Trait mindfulness differentiates the interest in healthy diet from orthorexia nervosa.

BACKGROUND: Obsessive healthy eating and its extreme form orthorexia nervosa are epidemiologically significant problems. Mindfulness, the focused, non-judgmental attention to and awareness of present events, may be an important psychological contributor to (orthorexic) eating habits. METHODS: In this cross-sectional survey-based study, 314 women and 75 men (mean agetotal sample = 27.17 years, SD = 10.64) provided data on mindfulness (Freiburg Mindfulness Inventory, presence and acceptance subscale) and orthorexic eating (Teruel Orthorexia Scale, healthy orthorexia and orthorexia nervosa subscale). RESULTS: In this study, we found a positive relation between mindfulness and healthy orthorexia, the non-pathological interest in eating healthy. By contrast, orthorexia nervosa, the pathological obsession with healthy eating, was negatively associated with mindfulness. Gender differences appeared neglectable. CONCLUSION: Taken together, these results confirm previous research showing that mindfulness encourages eating healthy and may protect against eating-related pathologies. Result also support the notion that orthorexia has two dimensions, healthy and nervosa, which are differently related to psychological factors, herein mindfulness. LEVEL OF EVIDENCE: Level III, cohort study.

Food cue-elicited brain potentials change throughout menstrual cycle: Modulation by eating styles, negative affect, and premenstrual complaints.

BACKGROUND: While there is evidence for increased food intake and craving during the luteal phase, underlying mechanisms are incompletely understood. The present study investigated electrophysiological responses to food pictures as a function of menstrual cycle phase. In addition, the moderating effects of progesterone, eating behaviors (restraint, emotional, orthorexic), negative affect, and premenstrual complaints were explored. METHODS: Using a within-subject design, 35 free-cycling women watched and rated pictures of food (high and low caloric) and control items during the follicular, the ovulatory, and the luteal phase (counterbalanced), while EEG was recorded to examine the late positive potentials (LPP). Salivary gonadal hormones and affect were examined at each occasion. Eating behaviors and premenstrual complaints were assessed once. RESULTS: For parietal regions, average LPPs were comparable between cycle phases but slightly larger LPP amplitudes were elicited by high caloric food pictures as compared to the neutral category. Descriptively, both food categories elicited larger parietal LPPs than neutral pictures during the luteal phase. Analyses of LPPs for central-parietal regions showed no effect of picture category or cycle phase, except higher amplitudes in the right area during the luteal phase. During the luteal phase, progesterone and functional interference from premenstrual symptoms (but not age, BMI, picture ratings, affect, estradiol, or eating behaviors) significantly predicted larger parietal LPPs towards high caloric (but not low caloric) pictures. CONCLUSION: Our findings suggest a heightened food cue reactivity during the luteal phase, which may relate to higher ovarian hormone secretion and more functional impact of premenstrual symptoms. This research contributes to a better understanding of menstrual health and the identification of preventive strategies for premenopausal women.

Neural correlates of gender differences in distractibility by sexual stimuli.

Attentional interference control is a prominent feature of human cognition. To what extent sexual stimuli attract attention and interfere with cognitive tasks has still little been studied. Our study aimed to identify associations between attentional interference, sexual arousal, trait sexual motivation, and neural activity to sexual distractors while accounting for gender differences. Therefore, the present study examined the neural correlates of attentional interference by arousing sexual distractors using functional magnetic resonance imaging (fMRI). Fifty women and 47 men underwent fMRI while indicating the orientation of two lines (equal or unequal) next to an explicit sexual (as compared to a neutral) picture. Results confirmed prolonged response times when a sexual image was shown. There was neither a difference between genders nor an effect of sexual arousal ratings or trait sexual motivation on distractibility. Neural activity specific to sexual images was found in brain regions implicated in motivation and reward processing. Men as compared to women showed stronger responses in the nucleus caudatus, the anterior cingulate cortex, and the nucleus accumbens. Trait sexual motivation was selectively correlated with nucleus caudatus activity. Taken together, findings support the notion that even when not in the focus, sexual images activate the brains' reward circuitry. Men's higher sensitivity to the rewarding value of sexual cues may be critical for their higher risk of addictive/compulsive sexual behaviors.

A silent, neutral substitution detected by reverse-phase high-performance liquid chromatography: hemoglobin Beirut.

A substitution of alanine for valine at position 126 in the beta-chain of hemoglobin was discovered in a hematologically normal adult male of Lebanese extraction. The variant beta-globin was initially observed and subsequently purified by reverse-phase high-performance liquid chromatography (HPLC). Reverse-phase HPLC was also used to isolate the variant tryptic peptide of beta-T13 that has alanine replacing valine at residue 126. The discovery of hemoglobin Beirut illustrates the usefulness of reverse-phase HPLC for the detection of neutral amino acid substitutions in proteins. The ability to detect neutral substitutions in undigested proteins is pertinent to the monitoring of genetic variation in human populations.

Attentional bias toward and distractibility by sexual cues: A meta-analytic integration.

From an evolutionary perspective, sexual stimuli are highly salient and are assumed to be processed with high priority. Hence, attentional processing of sexual cues is expected to not only bias attention but to also distract from other cognitive (foreground) tasks. It is, however, unclear to what extent these stimuli capture attention and whether there are differences between men and women. This meta-analysis combined the results of 32 studies employing experiments of attentional bias toward and distraction by sexual stimuli. From these, 13 studies provided data to examine gender differences. Overall, attentional bias and distractibility was lower than anticipated (gz = 0.43, p < .001) and there was support for the assumption of higher attention bias/interference in men (gs = 0.29, p = .031). Importantly, there was evidence for the presence of publication bias. With this in mind, findings are discussed in the context of stimulus features, the impact of provoked sexual arousal and motivational state, and gender-specific and -nonspecific neural processing of sexual stimuli which influence attention toward them.

Diurnal cortisol and alpha-amylase in the daily lives of older adults with vital exhaustion.

OBJECTIVE: Vital exhaustion (VE) is characterised by unusual fatigue, increased irritability, and a feeling of demoralisation. It has been found a major risk factor for cardiovascular diseases, and one that is independent of subclinical or clinical manifestations of coronary heart disease or lifestyle-related risk factors. Stress-induced alterations in the hypothalamic-pituitary-adrenal (HPA) axis and sympathetic nervous system may mediate the link between VE and increased cardiovascular risk. However, no studies have yet assessed both systems simultaneously and in high-risk populations, such as older adults. METHODS: A total of 72 older adults (34 women, mean age 61.7±7.3) who were free of any major physical or mental illnesses filled out the Maastricht Vital Exhaustion Questionnaire (MVEQ) and the Perceived Stress Scale (PSS). To determine cortisol and alpha-amylase, participants collected saliva samples upon awakening, +30min thereafter, and at 11am, 3pm, and 8pm. RESULTS: Participants with higher VE reported lower perceived stress (ß=-0.515, p<0.001). Individuals reporting higher VE also exhibited more diminished cortisol concentrations across the day, although only by trend (ß=-0.218, p=0.092). There was no significant association between VE and diurnal alpha-amylase activity. Moreover, women had lower diurnal cortisol (ß=-0.381, p=0.004) and alpha-amylase (ß=-0.329, p=0.011) when compared to men. CONCLUSIONS: Our findings provide initial evidence for psychosocial stress to be linked to VE in older adults, while evidence for HPA alterations remains tentative. Future research is warranted to determine whether VE related hypocortisolaemia represents a specific stage of the stress adaptation process that may put individuals at risk for incident cardiovascular diseases.

Differential expression of Op18 phosphoprotein during human thymocyte maturation.

Op18 (also termed prosolin/stathmin) is a highly conserved 18-kD cytosolic phosphoprotein expressed in low levels in mature resting G0 lymphocytes, but induced in late G1 and S phases after entry into the cell cycle. In addition to its induction in normal proliferating lymphocytes, Op18 has been found to occur at high levels in acute leukemias and in neuroendocrine tissue. The presence and rapid phosphorylation of Op18 after stimulation of proliferating cells correlates with subsequent functional responses of the cells, and, therefore, Op18 has been suggested to play a key role in signal transduction. The pattern of expression of Op18 during lymphoid development is of interest in view of its high levels of expression in acute leukemias, representing cells arrested at an immature stage, thus raising the possibility that Op18 may be regulated differently in mature and immature lymphoid cells. We report here that immature human thymocytes bearing the cortical double positive phenotype (CD4+CD8+) constitutively express high levels of Op18 protein. In contrast, in mature single positive thymocytes (CD3+CD4+ or CD3+CD8+), Op18 protein is expressed at a lower level, comparable to that seen in peripheral blood T cells. Cell cycle analysis demonstrated that most of the cells in the double positive thymocyte population expressing high levels of Op18 were noncycling and arrested in G0. Furthermore, there was no correlation between Op18 levels and the proportion of cycling cells in double positive thymocyte populations isolated from different thymuses. Interestingly, although Op18 protein levels did not increase any further after mitogenic stimulation of double positive thymocytes, an increase in Op18 phosphorylation was observed, thus coupling of Op18 phosphorylation to cell activation remained intact. Our results show that during lymphoid maturation Op18 expression is uncoupled from cell proliferation. These data also suggest that the ordered expression of proliferation-associated genes seen in mature T cells may be disrupted during T cell maturation.

Proliferation-related expression of p19/nm23 nucleoside diphosphate kinase.

High level expression of the nm23-H1 gene, which encodes for a nucleoside diphosphate kinase, has been found to correlate with diminished metastasis in some tumors but not in others. We have previously identified the protein product of the nm23-H1 gene in two-dimensional electrophoretic gels and have designated it p19/nm23. In neuroblastoma, higher levels of p19/nm23, which are associated with amplification of the N-myc oncogene, large tumor mass, and metastasis, were observed in advanced stage tumors compared with limited stage disease. Because of the variable expression of nm23-H1 in different tumors, we have investigated the relationship between amounts of the protein and cell proliferation. The levels of p19/nm23 were compared between resting and mitotically stimulated normal human PBLs and in leukemia cells. The amount of p19/nm23 increased in normal lymphocytes in response to mitotic stimulation and paralleled the increase in DNA synthesis. In leukemia cells obtained from patients with different subtypes of acute leukemia, p19/nm23 levels were also increased relative to resting normal lymphocytes. Treatment of mitotically stimulated lymphocytes with cyclosporin, which inhibits proliferation, blocked the increase in p19/nm23; treatment of the leukemia cell line HL-60 with dimethylsulfoxide, which induces terminal differentiation, resulted in diminished levels of p19/nm23. Our data therefore provide evidence that nm23-H1 expression is related to cell proliferative activity.

Identification of two related markers for common acute lymphoblastic leukemia as heat shock proteins.

By direct analysis of the polypeptide constituents of leukemic cells, we have previously detected several polypeptides that are restricted in their expression to acute lymphoblastic leukemia (ALL). In this study, we provide evidence that two polypeptides designated L2 and L4 are structurally related and represent novel markers for common ALL. Partial amino acid sequence analysis did not uncover differences between L2 and L4. The sequences obtained correspond to a previously cloned human gene designated hsp 27 that is expressed, following heat shock treatment, in a variety of cells. 32Pi incorporation studies indicate that L4 is an unphosphorylated form and L2 is a phosphorylated form of hsp27. The two forms were inducible by heat shock in leukemic and nonleukemic lymphoid cells. Thus, in acute leukemia, the common ALL subtype is uniquely characterized by the constitutive expression of a polypeptide that represents a major cellular phosphoprotein.

High levels of p19/nm23 protein in neuroblastoma are associated with advanced stage disease and with N-myc gene amplification.

The gene encoding a novel protein designated nm23-H1, which was recently identified as identical to the A subunit of nucleotide diphosphate kinase from human erythrocytes, has been proposed to play a role in tumor metastasis suppression. We report that untreated neuroblastoma tumors contain a cellular polypeptide (Mr = 19,000) designated p19, identified in two-dimensional electrophoretic gels, which occurs at significantly higher levels (P = 0.0001) in primary tumors containing amplified N-myc gene. The partial amino acid sequence obtained for p19 is identical to the sequence of the human nm23-H1 protein. An antibody to the A subunit of erythrocyte nucleotide diphosphate kinase reacted exclusively with p19. In this study, significantly higher levels of p19/nm23 occurred in primary neuroblastoma tumors from patients with advanced stages (III and IV) relative to tumors from patients with limited stages (I and II) of the disease. Even among patients with a single copy of the N-myc gene, tumors from patients with stages III and IV had statistically significantly higher levels of p19/nm23 than tumors from patients with stages I and II. Our findings indicate that, in contrast to a proposed role for nm23-H1 as a tumor metastasis suppressor, increased p19/nm23 protein in neuroblastoma is correlated with features of the disease that are associated with aggressive tumors. Therefore, nm23-H1 may have distinct if not opposite roles in different tumors.

Hsp27 expression in neuroblastoma: correlation with disease stage.

BACKGROUND: The 27-kd heat shock protein (Hsp27) is differentially expressed in some malignancies, including breast carcinoma, leukemia, and malignant fibrous histiocytoma. In breast carcinoma, a high-level expression of Hsp27 has been associated with shorter disease-free survival in patients with localized disease. PURPOSE: We have observed variable levels of Hsp27 among neuroblastoma tumors. Our aim in this study was to investigate the relationship between Hsp27 expression and stage of the disease and N-myc gene copy number. METHODS: We determined Hsp27 protein levels in 53 neuroblastoma tumors representing different stages of the disease and in 17 neuroblastoma cell lines by quantitative two-dimensional polyacrylamide gel electrophoresis (PAGE). We also performed statistical analysis of Hsp27 levels in relation to stage of the disease and to N-myc gene copy number. RESULTS: Increased Hsp27 expression in neuroblastomas was associated with limited stage disease and inversely correlated with N-myc gene amplification, a feature known to predict poor clinical outcome. An inverse correlation was also observed between N-myc gene amplification and Hsp27 protein levels among the neuroblastoma cell lines analyzed. Immunohistochemical staining of sections of neuroblastomas showed that Hsp27 was most prominently expressed in the cytoplasm of large ganglionic tumor cells present in neuronally differentiated areas of the tumors. Induction of neuronal differentiation in SMS-KCNR neuroblastoma cells using retinoic acid resulted in an increase in Hsp27. CONCLUSION: High level expression of Hsp27 in neuroblastoma is a feature of limited stage, differentiated tumors. IMPLICATION: Hsp27 may play a part in the biology of neuroblastomas with a favorable outcome.

A nucleoside diphosphate kinase A (nm23-H1) serine 120-->glycine substitution in advanced stage neuroblastoma affects enzyme stability and alters protein-protein interaction.

A high level of nucleoside diphosphate kinase A (NDPK A/nm23-H1) in neuroblastoma is associated with advanced stage disease. We have also found a serine 120-->glycine substitution in NDPK A and/or amplification of the nm23-H1 gene in advanced stage neuroblastomas. Serine 120, a highly conserved residue, is located in proximity to histidine 118 which forms a phosphorylated intermediate essential for NDPK activity. The effect of Ser120-->Gly substitution on the biochemical properties of NDPK A was investigated. Phosphate-transferase activity was lower in the recombinant mutant NDPK A and in the immunoprecipitated complex consisting of NDPK A and NDPK B prepared from a neuroblastoma tumor containing the mutation, relative to the wild-type. There was a significant decrease in the enzyme stability toward urea- or temperature-induced denaturation for the recombinant mutant NDPK A and in an immunoprecipitate from a tumor containing the mutation. Recombinant NDPK A containing the Ser120-->Gly mutation exhibited reduced hexameric and increased dimeric oligomerization relative to the wild-type. Moreover a 28 kDa cellular protein was detected, that co-precipitated with the mutant but not wild-type NDPK A. The altered properties of the mutant protein may have relevance to a role for NDPK A in neuroblastoma progression.

N-myc gene amplification in neuroblastoma is associated with altered phosphorylation of a proliferation related polypeptide (Op18).

We have recently identified and cloned the gene for a cytosolic polypeptide, designated oncoprotein 18 (Op18), which is expressed in acute lymphocytic leukemia and some solid tumors including neuroblastoma. Op18 is phosphorylated upon treatment of lymphoid cells with phorbol myristate acetate. We have proposed that unphosphorylated Op18 plays a role in cellular proliferation, and that its phosphorylated forms, namely Op18a and Op18b, are associated with diminished cell proliferation. In this study, we report that in neuroblastoma tumors, the phosphorylation of Op18 was substantially diminished with increasing N-myc gene copy number. Treatment of the neuroblastoma cell line SMS-KCNR, which contains 75 copies of the N-myc gene, with retinoic acid for ten days resulted in an increase in Op18 phosphorylation. Our findings provide evidence for distinct patterns of Op18 phosphorylation in neuroblastoma tumors with and without N-myc gene amplification.

Two distinct pancreatic amylase genes are active in YBR mice.

The genetic determinants of pancreatic amylase expression in YBR mice differ in two respects from those of other inbred strains. First, there are two nonallelic amylase isozymes present in YBR pancreas, while most mouse strains express a single pancreatic amylase protein. In addition, the in vivo rate of total pancreatic amylase synthesis is 50% of that in other strains. Both these traits are determined by genetic sites in the region of the Amy-2 locus on mouse chromosome 3. To determine the molecular basis for the presence of two isozymes in this strain, we have compared portions of their amino acid sequences. Two differences between isozymes A1 and B1 were identified among the 77 residues compared. This result demonstrates that two distinct amylase genes are expressed in YBR pancreas.

Identification of novel Escherichia coli ribosome-associated proteins using isobaric tags and multidimensional protein identification techniques.

Biogenesis of the large ribosomal subunit requires the coordinate assembly of two rRNAs and 33 ribosomal proteins. In vivo, additional ribosome assembly factors, such as helicases, GTPases, pseudouridine synthetases, and methyltransferases, are also critical for ribosome assembly. To identify novel ribosome-associated proteins, we used a proteomic approach (isotope tagging for relative and absolute quantitation) that allows for semiquantitation of proteins from complex protein mixtures. Ribosomal subunits were separated by sucrose density centrifugation, and the relevant fractions were pooled and analyzed. The utility and reproducibility of the technique were validated via a double duplex labeling method. Next, we examined proteins from 30S, 50S, and translating ribosomes isolated at both 16 degrees C and 37 degrees C. We show that the use of isobaric tags to quantify proteins from these particles is an excellent predictor of the particles with which the proteins associate. Moreover, in addition to bona fide ribosomal proteins, additional proteins that comigrated with different ribosomal particles were detected, including both known ribosomal assembly factors and unknown proteins. The ribosome association of several of these proteins, as well as others predicted to be associated with ribosomes, was verified by immunoblotting. Curiously, deletion mutants for the majority of these ribosome-associated proteins had little effect on cell growth or on the polyribosome profiles.

Advances in two-dimensional electrophoresis.

New developments in the first dimension step of two-dimensional electrophoresis have expanded the utility of the technique in cell and molecular biology.

Identification and characterization of a human transthyretin variant.

An apparent Mr variant of plasma transthyretin (TTR), previously detected using 2-D PAGE, is the first reported occurrence of this type of human TTR variant. We characterized the variant TTR to determine the nature of this difference. Comparative tryptic peptide maps of variant and normal TTR and sequencing of peptides which differed indicated the variant contained a single amino acid substitution of valine for tyrosine at position 116. Because such a change requires two nucleotide substitutions, we postulate the variant arose through mutation in codon 116 of a heretofore unrecognized polymorphic or rare variant allele of TTR.

Moving and stationary boundaries in immobilized pH gradients.

The relations describing the concentration changes at moving boundaries in a medium containing bound, buffering group are derived for a system which, except for hydrogen and hydroxyl ions, contains one anionic and one cationic mobile constituent. The relations found have been used to calculate concentrations and conductivities in zones developing in immobilized pH gradients. Assumptions used in the calculations as well as conductivity ratios between zones have been experimentally controlled and were found to reasonably agree with expectations. It is also shown how difference in transference numbers between sample droplet and gel will cause concentration and pH changes at the gel-sample droplet interfaces and it is explained how these changes are related to ionic concentrations in the gel. The high concentration zone generated at one of the interfaces will be transported into the gel. This transport has been numerically simulated and experimentally verified. The low concentration generated at the opposite interface will cause titration impeding sample entrance in the gel through this interface even when the gel contains ions other than H+ or OH- transported towards the interface. The described phenomena explain the dependence of lateral spreading, precipitation at the application site as well as streaking and smearing along sample lanes, on the type and concentration of low molecular weight ions originally present in the gel.

Somatic mutation studies in human lymphoid cells: the detectability of quantitative genetic variants in two-dimensional gels.

The feasibility of detecting quantitative genetic variants based on a decrease in the integrated intensity of polypeptide spots in two-dimensional polyacrylamide gels of human lymphoblastoid cell clones was investigated. A battery of 65 spots on 115 gels was studied to determine the distribution of quantitative measures for spots where no mutation had occurred. The corresponding distribution for spots which have decreased integrated intensity as a result of a mutation at one of two alleles coding for the spot was investigated by quantitating spots for which mutations were known to have occurred. These two distributions allowed the estimation of the rates of false positive and false negative errors for any particular strategy aimed at detecting null mutations, and thus provides a basis for the design of efficient strategies. Our silver stained gels have sufficient reproducibility of spot integrated intensities so that, for situations in which the mutation rate is relatively high, it is practical to monitor a subset of spots for null variants using the same digitized images as are used to detect structural variants.

Pigmentation and lysosomal phenotypes in mice doubly homozygous for both light-ear and pale-ear mutant alleles.

We have developed a new strain of mice homozygous for mutant alleles at both the light-ear locus on chromosome 5 and the pale-ear locus on chromosome 19. The pigmentation pattern of the double mutants, designated light-pale, is indistinguishable from the parental type. Elevated concentrations of lysosomal enzymes observed in certain tissues of the light-ear and pale-ear singly homozygous mice also are present in the double mutants, and are quantitatively indistinguishable from either parent. Although both mutations have pleiotropic effects on organelles in several tissues, neither locus influences the secretion of pancreatic zymogen granules. The close similarity in phenotypes of light ear, pale ear, and light-pale mice suggest that the le and ep loci encode different subunits of a multimeric protein, and that mutations affecting either subunit result in comparable losses of function.