Structure-permeation relations of met-enkephalin peptide analogues on absorption and secretion mechanisms in Caco-2 monolayers

Due to the low effective permeabilities of peptides at many absorption sites, their structure-permeation relations are of high interest. In this work structure-permeation relations of Met-enkephalin analogues are presented using confluent Caco-2 cells as an in vitro permeation model. Four model peptides (Met-enkephalin, [D-Ala2]Met-enkephalin, [D-Ala2]Met-enkephalinamide, and metkephamid) were tested in terms of permeability, lipophilicity, charge, and molecular size. Permeability coefficients (P(eff)) across Caco-2 cells were low, 3.3 x 10(-8) to 9.5 x 10(-8) cm s-1, and were similar to typical paracellular markers. No correlation of permeability and the log(apparent octanol/buffer partition coefficient) was observed. A 40-fold increase of the permeability of metkephamid in the presence of 10 mM EDTA suggested a significant contribution of paracellular transport. Independent support for this conclusion was obtained by visualizing the pathway of the fluorescein isocyanate isomer I 1-metkephamid by confocal laser scanning microscopy (CLSM). The fluorophore-labeled peptide was observed in the intercallular space only. Metkephamid permeabilities were found to be direction-specific. Permeabilities from basolateral to apical (b-to-a) were significantly higher (ca. 4-fold) than in the opposite (a-to-b) direction. The addition of verapamil equalized the permeabilities in the a-to-b and b-to-a directions, suggesting the involvement of a P-glycoprotein-mediated secretion mechanism. Similar observations were obtained with [D-Ala2]Met-enkephalinamide, but not with Met-enkephalin and [D-Ala2]Met-enkephalin. In contrast to the other analogues, metkephamid and [D-Ala2]Met-enkephalinamide are positively charged at neutral pH, as demonstrated by their isoelectric points (pl = 8.6 for [D-Ala2]Met-enkephalinamide and metkephamid and 5.3 for [D-Ala2]Met-enkephalin and Met-enkephalin). The data is in agreement with the literature showing that most compounds secreted by the P-glycoprotein transporter carry a positive charge.     

An artificial intelligent diagnostic system on differential recognition of hematopoietic cells from microscopic images

The endothelium participates actively in homeostatic mechanisms such as the regulation of vascular tone and maintenance of a nonthrombotic environment, as well as directing biological responses such as leukocyte trafficking to inflammatory sites. Disruption of these processes leads to disease. In the antiphospholipid antibody syndrome autoantibodies provoke the endothelium to develop a prothrombotic surface. In systemic vasculitides associated with presence of antineutrophil cytoplasm antibodies, it is likely that the autoantibodies incite premature neutrophil activation, disrupted neutrophil-endothelium interactions and endothelial damage. This review considers how normal endothelial functions may be subverted in disease and how active endothelial responses may contribute to disease.     

Thermally-induced changes in the metabolism of the eye of the crayfish Paranephrops planifrons: respiration and substrate utilization at three distinct temperatures

Gleason's score (GS) has been reported to be the most valuable prognostic factor in cases of prostate cancer. GS is solely dependent on the histological architecture of the prostate cancer, but, it seems doubtful that histological patterns are sufficient for evaluating the degree of malignancy of prostate cancer. We previously reported that the estimation of volume-weighted mean nuclear volume (MNV) might be a more useful prognosticator for prostate cancer than subjective histological grading. However, the previous study was conducted on patients treated in a single hospital, and the number of subjects was too small to draw a definitive conclusion. In this study, we analyzed a larger number of subjects at another institution using a blinded study design. A retrospective prognostic study of 195 patients with prostate cancer diagnosed between January 1966 and December 1988 at Kyoto University Hospital, and treated by conservative therapy, was conducted. Unbiased estimates of MNV were compared with the clinical stage and histological grading according to GS with regard to the prognostic value. Univariate analysis revealed that estimates of MNV, clinical stage, and GS all correlated significantly with disease-specific survival in cases of prostate cancer. Multivariate analysis of all cases also revealed that all of these factors were significant independent prognosticators of disease-specific survival. However, focusing on clinically localized cases (stages A, B, and C), multivariate analysis revealed that the estimation of MNV was the only powerful prognosticator of prostate cancer. This study indicates that the estimation of MNV is prognostically equal or superior to GS in cases of prostate cancer. We emphasized that the estimates of MNV is a more objective method for histological grading to predict the malignant potential of prostate cancer.     

Physiologic correlates to running performance in pre-pubertal distance runners

In adults, four major variables have been shown to be associated with success in distance running performance: submaximal oxygen consumption (running economy), peak oxygen consumption (Peak VO2), ventilatory threshold (VT) and fractional utilisation (FU). The primary aim of this study was to describe the relationship between the 3000 m race times of run-trained prepubertal boys to these four variables. Thirteen male run-trained pre-pubertal boys (age 11.7 +/- 1.1 yrs, mean +/- SD), volunteered to take part in a 3000 m time trial and laboratory assessment, consisting of treadmill running at four submaximal speeds (8, 9.6, 11.2 and 12.8 km.h-1) as well as a peak VO2 test. The group demonstrated a heterogeneous array of peak VO2 data. A high level of association (p < 0.05) was found between mass-relative peak VO2 and 3000 m time trial results (r = -0.83). In addition ventilatory threshold expressed as %peak VO2, VO2 at VT and estimated velocity at VT was also highly related to 3000 m time trial (r = -0.78, -0.77 and -0.77) respectively. Fractional utilisation (%peak VO2) was significantly (p < 0.05) associated with race time at the final two submaximal running speeds only (11.2 and 12.8 km.h-1) (r = 0.61 and 0.67, respectively). Respiratory Exchange Ratio (RER) was also found to be significantly (p < 0.05) associated with 3000 m race time at 11.2 and 12.8 km.h-1. Overall peak VO2 appeared to be the single most important factor associated with success at 3000 m.     

Correlates of alcohol use and misuse in fourth-grade children: psychosocial, peer, parental, and family factors

To determine level of alcohol use/misuse and to examine correlates of these behaviors, 1,314 fourth-grade students were surveyed. The questionnaire included 55 items concerning tolerance of deviance, deviant self-image, self-efficacy, susceptibility to peer pressure, personal and peer approval of alcohol use, peer adjustment, parent nurturance and monitoring, family adjustment, parental permissiveness, peer use of alcohol, and exposure to alcohol. The items were factor analyzed and indices constructed. The indices generally had acceptable alpha coefficients (alpha = .61-.91); two exceptions were peer adjustment (alpha = .51) and parental permissiveness (alpha = .42). Tolerance of deviance, deviant self-image, susceptibility to peer pressure, personal and peer approval, peer use and exposure by peers, and parental permissiveness were positively correlated with alcohol use/misuse. Self-efficacy, child-parent interactions, family adjustment, and peer adjustment were negatively correlated with alcohol use/misuse. Implications for the design of family-based alcohol use/misuse prevention programs are discussed.     

Cognitive distortions and adolescent affective disorder. Validity of the CNCEQ in an inpatient sample. Children's Negative Cognitive Error Questionnaire

Despite a proliferation of recent research examining childhood and adolescent depression, the area still lags behind the adult depression field, particularly in the investigation of cognitive correlates of affective psychopathology. To advance cognitive research with youth, the Children's Negative Cognitive Error Questionnaire (CNCEQ) was developed to provide a measure of cognitive distortions or errors in children and adolescents. Yet, few studies have employed the CNCEQ and no evidence exists supporting the validity of its four component cognitive error scales. The purpose of the present study was to examine the construct validity of the CNCEQ and its constituent scales through the use of factor analysis and criterion-group comparisons. Groups of adolescent psychiatric inpatients, diagnosed as affective or disruptive disordered, completed the CNCEQ following admission. Results failed to support the implicit four-factor structure of the CNCEQ, instead suggesting the appropriateness of a single-factor solution labeled "negative thinking." Despite no diagnostic group differences on the CNCEQ total or other scale scores, affective disordered patients evinced more cognitive errors on the Overgeneralizing scale. Findings suggest that the CNCEQ in its current stage of development holds promise, yet requires refinement to produce a valid measure of cognitive functioning in youth.