Caregiver education in Parkinson's disease: formative evaluation of a standardized program in seven European countries.

PURPOSE: The formative evaluation of a standardized psychosocial education program for patients with Parkinson's disease (PD) and their caregivers. The results of the participation of the caregivers are presented next to the data of the patients. METHODS: Caregivers (n = 137) and patients with PD (n = 151) participated in the 8-week program in separate groups. Measurements were performed on psychosocial problems (BELA-P/A-k), health state (EQ-5D VAS), quality of life (PDQ-39) and depression (SDS) 1 week before and 1 week after the program. Participants rated their mood on a visual analogue scale before and after each session, and they filled in an evaluation questionnaire after the last session. RESULTS: Scores on the BELA-P/A-k improved significantly on the 'bothered by scale' as well as the 'need for help scale'. No improvements were found on EQ-5D VAS, PDQ-39 and SDS. Mood ratings improved significantly after each session. Most participants evaluated the program as positive. Feedback led to improvements in the program, which are incorporated in a final manual. CONCLUSIONS: The program was feasible to run in the different countries. This exploratory study led to improvements in the program and recommendations for further research. A study on the effectiveness of the program is the next step.

Patterns of psychological problems in Parkinson's disease.

OBJECTIVE: To identify patterns of psychological problems in Parkinson's disease (PD). METHOD: A sample of 3075 patients was surveyed to determine frequencies of psychological problems and cross-validated cluster analyses were computed to identify patterns of these problems. RESULTS: An increase of symptoms during arousal was reported by 68%, sleep disturbances by 32% of the sample, and sexual problems by 57% of men and 22% of women. Less frequently reported were difficulties in communicating (27%), needing help of others (38%), and depressive moods (20%). Four patterns of psychological problems were identified: general low stress, general high stress, sexual and social problems, and non-social problems. CONCLUSION: The study confirms the clinical observation that PD patients differ not only in degree, but also in structure of psychological stress. Social and non-social stress constitute principal types of stress experienced in PD. This distinction should be taken into account for any approach to support people with PD.

Chocolate eating in healthy men during experimentally induced sadness and joy.

The study compared influences of qualitatively different emotions on eating. Motivation to eat, affective responses to chocolate and chewing of chocolate were investigated in healthy normal weight males during experimentally induced emotions. Subjects abstained from eating 2 h (n = 24) or 8 h (n = 24) before testing. They received pieces of chocolate after viewing film clips presented to induce anger, fear, sadness and joy. Motivation to eat and most affective responses to eating chocolate were higher after 8 h than after 2 h of deprivation. Sadness and joy affected motivation to eat in opposite directions: joy increased and sadness decreased appetite (p < 0.001). In joy, a higher tendency to eat more chocolate was reported (p < 0.001), and chocolate tasted more pleasant (p < 0.001) and was experienced as more "stimulating" than in sadness (p < 0.01). No effects of deprivation could be found for chewing time and number of chews. Results indicate that the quality of emotions can affect motivation to eat and affective responses to eating chocolate. Our findings on decreased eating responses to sadness in healthy males and the contradictory increased eating responses to sadness reported by others supports two types of emotion-induced changes of eating: emotion-congruent modulation of eating and eating to regulate emotions.

Molecular structure, processing, and tissue distribution of matrilin-4.

Matrilin-4 is the most recently identified member of the matrilin family of von Willebrand factor A-like domain containing extracellular matrix adapter proteins. Full-length matrilin-4 was expressed in 293-EBNA cells, purified using affinity tags, and subjected to biochemical characterization. The largest oligomeric form of recombinantly expressed full-length matrilin-4 is a trimer as shown by electron microscopy, SDS-polyacrylamide gel electrophoresis, and mass spectrometry. Proteolytically processed matrilin-4 species were also detected. The cleavage occurs in the short linker region between the second von Willebrand factor A-like domain and the coiled-coil domain leading to the release of large fragments and the formation of dimers and monomers of intact subunits still containing a trimeric coiled-coil. In immunoblots of calvaria extracts similar degradation products could be detected, indicating that a related proteolytic processing occurs in vivo. Matrilin-4 was first observed at day 7.5 post-coitum in mouse embryos. Affinity-purified antibodies detect a broad expression in dense and loose connective tissue, bone, cartilage, central and peripheral nervous systems and in association with basement membranes. In the matrix formed by cultured primary embryonic fibroblasts, matrilin-4 is found in a filamentous network connecting individual cells.

Mass spectrometric analysis of head-to-tail connected cyclic peptides.

Tandem mass spectrometry is an extremely useful tool for high sensitive sequence identification of peptides. In the case of cyclic peptides fragmentation can easily be performed for sequence analysis. However, analysis is usually tedious due to the lack of a defined beginning and end of the sequence. Since cyclic peptides are a highly interesting class of compounds especially for the pharmaceutical industry, ways have to be found to identify their strictures. In this work we demonstrate how software and dedicated analytical strategies can be used for detailed analysis of these substances.

Magnetic, microplate-format plasmid isolation protocol for high-yield, sequencing-grade DNA.

We have developed a rapid, microplate-format plasmid isolation procedure to purify sequencing-grade DNA templates for high-throughput DNA sequencing operations. A modified lysozyme/boiling method is used to produce a plasmid-containing supernatant that is then purified by iron bead capture. After binding, the beads are pelleted in a magnetic field, washed and the DNA eluted in water. The method yields up to 10 micrograms plasmid DNA from a 1-mL overnight culture in a deep-well microplate. The procedure is suitable for large-scale experiments, amenable to automation and does not require expensive reagents or equipment. The entire protocol can be completed in as little as 2 h, and one technician with a 96-well pipetting station can process up to 48 plates per day. This protocol is ideal for any high-throughput operation in which template quantity, quality and reproducibility are of primary importance.

Emotions and eating in everyday life.

This field study assessed emotional states experienced in everyday life and examined the subjective motivation to eat associated with these emotional states. Twenty-three female subjects rated their momentary emotional state and motivation to eat on 6 consecutive days at 11:00a.m., 2:00p.m., 5:00p.m., 8:00p.m. and 11:00p.m. A cluster analysis of the resulting 634 emotion profiles revealed three types of emotional states characterized by the labels "Anger-dominance", "Tension/Fear" and "Relaxation/Joy". A fourth cluster showing generally low levels of emotions was labelled "Unemotional state". Most of the self-rated motivations to eat were increased during periods of negative emotions. During negative emotions a heightened tendency to cope with these emotions through eating and more intense bodily symptoms of hunger were also reported. No differences in motivations to eat were found between the two negative emotion clusters or between relaxation/joy and the unemotional state. Results indicate the presence of "emotionally instrumental eating" in a non-clinical population under real life conditions. Physiological correlates of negative emotional states may be involved in emotionally instrumental eating.

Characteristics of eating in anger, fear, sadness and joy.

The influences of emotions on eating were investigated. One-hundred and seven female and 103 male subjects were asked to report how various characteristics of eating could change during the emotions of anger, fear, sadness and joy. Subjects completed a questionnaire of 33 items for each emotion and a food deprivation condition. A factor analysis of the items answered with respect to food deprivation yielded four factors which were labelled hunger, impulsive eating, sensory eating and hedonic eating. ANCOVAs showed main effects of emotions upon each factor. Single comparisons showed that: (1) subjects reported to experience higher levels of hunger during anger and joy than during fear and sadness; (2) impulsive eating and sensory eating were rated higher during anger than during the other emotions; and (3) hedonic eating was rated higher during joy than during the other emotions. Results did not indicate any differential influences upon eating between fear and sadness. Women reported to experience higher tendencies of impulsive eating and sensory eating than men during anger and sadness. Eating scores of negative emotions correlated low and positive with body mass index, low and negative with dietary restraint. The differential effects of emotions on characteristics of eating may be due to differences of the examined emotions in frequency of occurrence, physiological correlates and motivational properties.

Behavioral analysis of the freezing phenomenon in Parkinson's disease: a case study.

The "freezing" phenomenon was examined in a 77 yr old male patient suffering from Parkinsons's disease (PD). Motor blocks occurred several times a day, mainly affected walking, and lasted mostly between 2 and 5 min. Behavioral analysis showed that freezing was elicited and intensified not only by external factors (limited space and room to move), but also by negative cognitions, negative emotions and tension. The most effective behavior to overcome freezing was a combination of relaxation and loud rhythmic counting. Effectiveness in overcoming freezing correlated positively with self-rated mobility before trials. Relaxation training improved self-rated mobility. It is suggested that relaxation training and cognitive techniques should be used to reduce tension, negative thoughts and negative emotions contributing to freezing and to improve the ability to cope with freezing.

Correlation between chlamydial infection and autoimmune response: molecular mimicry between RNA polymerase major sigma subunit from Chlamydia trachomatis and human L7.

L7 is one of the ribosomal proteins frequently targeted by autoantibodies in rheumatic autoimmune diseases. A computer search revealed a region within the immunodominant epitope of L7 (peptide II) that is highly homologous to amino acid sequence 264-286 of the RNA polymerase major sigma factor of the eubacterium Chlamydia trachomatis. Anti-L7 autoantibodies affinity purified from the immunodominant epitope were able to recognize this sequence as they reacted with purified recombinant sigma factor. Immunofluorescence labeling experiments on C. trachomatis lysates revealed a punctate staining pattern of numerous spots when incubated with the affinity-purified anti-peptide II autoantibodies. Binding of autoantibodies to peptide II was inhibited by the homologous sigma peptide. This is the first demonstration of epitope mimicry between a human and a chlamydial protein on the level of B cells. Antibody screening revealed a significant correlation between the presence of anti-L7 autoantibodies and C. trachomatis infection in patients with systemic lupus erythematosus and mixed connective tissue disease. Our results suggest that molecular mimicry is involved in the initiation of anti-L7 autoantibody response and may represent a first glance into the immunopathology of Chlamydia with respect to systemic rheumatic diseases.

Molecular characterization of a conformational epitope of hen egg white lysozyme by differential chemical modification of immune complexes and mass spectrometric peptide mapping.

A new approach for the characterization of conformationally dependent epitope structures in protein antigens is described using differential chemical modification of immune complexes in combination with mass spectrometric peptide mapping analysis. Well-established methods for epitope characterization are frequently not applicable to conformationally dependent epitopes, and direct methods of structure analysis such as X-ray crystallography of immune complexes have been successful only in a few cases. Our approach combines tertiary structure-selective chemical modification of immune complexes with the molecular characterization of reaction products by mass spectrometric peptide mapping. The comparison of the modification pattern of free and antibody-bound antigen provides the identification of residues protected from modification by the antibody. These residues hence are characterized as part of the epitope structure. The well-characterized hen egg white lysozyme and a corresponding monoclonal IgM-type antibody were investigated as a model system. Specific modification reactions for arginine, lysine, and tyrosine residues were performed, and the modification sites in free and antibody-bound antigen were determined by mass spectrometric peptide mapping. The R14 residue and residues K13 and K96 in the antibody-bound lysozyme were found to be protected from modification, comprising a surface of spatially adjacent residues by folding of the native protein. In contrast, other K and R residues as well as Y20 and Y23 showed no significant shielding from modification in the immune complex. These results provided an estimation of the molecular epitope surface area of native lysozyme.

Effects of Noise-Induced Arousal on Chewing of Sweet Food and the Subjective Motivation to Eat.

The effects of noise-induced arousal on chewing, subjective motivation to eat, the pleasantness of sweet food, and food intake were investigated. Twenty-four healthy normal weight male subjects received cookies differing in sucrose content (2%, 11%, 18% or 27%) before, during, and after 10 min of intermittent white noise. Half of the subjects were exposed to high-level noise (95 dB(A)) to induce psychological arousal. The remaining other half were exposed to low-level noise (60 dB(A)) as a control condition. During noise exposure the high-level noise group exhibited a shorter chewing time and fewer overall chewing movements with cookies of a medium sucrose content (11% or 18%). After exposure, subjects who received the high-level noise reported a reduced desire to eat, and tended to consume less strawberry ice cream. No significant effect of noise and Noise × Sugar interaction could be detected from taste and pleasantness ratings. The findings point to a relation between arousal, hunger motivation and chewing behavior.

Scanning behavior of rats during eating under stressful noise.

Behavioral ecological theories postulate that threatening environments should increase eating speed and vigilance during feeding. In the present experiment, eating speed and scanning behavior during eating were measured in 36 rats in 5 consecutive test sessions under stressful noise (95 dB white noise, n = 18) and control conditions (60 dB, n = 18) after the animals had been habituated to the test environment. Intense noise induced an increase of scanning rate and eating speed. These effects are similar to those reported for novel and light environments.

Mass spectrometric mapping of protein epitope structures of myocardial infarct markers myoglobin and troponin T.

Monoclonal antibodies are widely used analytical tools in biochemical research. The knowledge of their corresponding epitopes is of major interest. One possible approach for epitope characterization is the application of protein antigen proteolysis in combination with mass spectrometric peptide mapping analysis. Two complementary analytical strategies were applied: (a) limited proteolysis of antibody-bound antigen followed by removal of nonbound peptides and detachment of the antigenic peptides (epitope excision) and (b) enzymatic digest of the antigen followed by extraction of the antigenic peptides with the antibody and detachment of antigenic peptides after removal of nonbinding fragments (epitope extraction). In the few examples published so far, immobilized antibodies were used for these studies. In this study we present a method for characterization of the epitope sequences without prior immobilization of the monoclonal antibody. The separation of nonepitope peptides from antibody-bound peptides was carried out by ultrafiltration. The epitope and nonepitope fractions were analyzed by MALDI-MS without further purification, and the epitope sequences were identified. The method was developed using a model system consisting of the synthetic C-terminal cyanogen bromide fragment CB3 of myoglobin and the commercial monoclonal anti-myoglobin MG1. In further investigations the epitope sequence of a synthetic 32 amino acid peptide derived from heart muscle protein troponin T toward a monoclonal antibody MAb-M7, which was raised against the intact protein, was characterized. With this approach the epitope binding site of this antibody was determined, and selective shielding of potential cleavage sites in the immune complex could be observed. Furthermore, statements about the three-dimensional structure of the bound antigen were made.

Effects of stressful noise on eating and non-eating behavior in rats.

Eating and other behaviors were measured in 36 food-deprived rats on 15 consecutive days during 20 min test sessions. During training sessions 1 to 5 all animals were habituated to the test boxes with white noise of 55 dB intensity. For sessions 6 to 10 noise intensity was increased to 95 dB for the experimental rats and to 60 dB for the control rats. The food intake of experimental rats was lower for stress session 1. The duration of eating behavior was lower, and durations of exploring, grooming and resting behaviors were higher for all stress sessions for rats exposed to 95 dB white noise. Speed of eating behavior was higher for all stress sessions in the experimental group. Defecation rate of the experimental rats was higher for all stress sessions. On post-stress sessions 11 to 15 animals were again tested under the stimulus conditions of the training period (55 dB). No significant effects were observed for this period. The results are discussed with respect to models of "stress-induced" eating and behavioral ecology.

Effects of high- and low-energy meals on hunger, physiological processes and reactions to emotional stress.

The effects of a reduced energy content of two meals on hunger motivation, physiological variables and reactions to emotional stress were investigated. Healthy normal-weight male students received breakfast and lunch in the laboratory. Half of the subjects (n = 28) received meals with normal energy content (1700 kcal), and half received meals with reduced energy content (260 kcal). Psychological and physiological variables were obtained for 8 h from morning to afternoon, including during a period of emotional stress in the afternoon. Psychophysical state was altered by the reduction of energy in food (e.g. increased subjective motivation to eat, decreased systolic blood pressure). Noise decrease feelings of relaxation in subjects who had received low-energy meals, but not in subjects who had received high-energy meals. This enhanced emotional reactivity after low-energy intake is interpreted as a biologically meaningful consequence of the heightened hunger motivation. Furthermore, subjective hunger motivation was potentiated by stress when energy intake in the preceding hours was low. The latter result may be due to increased emotional reactions and/or an augmentation of deprivation-induced physiological changes by noise-induced emotional stress.