The Influence of Religious Affiliation on Participant Responsiveness to the Complete Health Improvement Program (CHIP) Lifestyle Intervention.

Seventh-day Adventist (SDA) and non-SDA (21.3 and 78.7 %, respectively) individuals (n = 7172) participating in the Complete Health Improvement Program, a 30-day diet and lifestyle intervention, in North America (241 programs, 2006-2012) were assessed for changes in selected chronic disease risk factors: body mass index (BMI), blood pressure (BP), pulse, lipid profile and fasting plasma glucose (FPG). Reductions were greater among the non-SDA for BMI, pulse and blood lipids. Furthermore, the majority of non-SDA in the highest risk classifications for BP, lipids and FPG, but only some lipids among SDA, were able to show improvement by 20 % or more.

The effects of an intensive lifestyle modification program on sleep and stress disorders.

BACKGROUND: To determine if a lifestyle change program can modify behavior to reduce sleep and stress disorders. METHODS: Analyses are based on 2,624 individuals aged 30 to 80 years from the Rockford, Illinois metropolitan area who completed a lifestyle evaluation at baseline and again after four weeks, following participation in a 40-hour educational course given over a four-week period. Participants receive instruction on the importance of making better lifestyle choices related to making long-term improvements in nutrition and physical activity and they learn ways to improve sleep and reduce stress in their lives. RESULTS: Significant percent decreases were observed in the number experiencing selected sleep or stress disorders from baseline to four weeks later for "sleeps restlessly" (-59%), "suffers from insomnia" (-64%), "feels under pressure" (-37%), "easily emotionally upset" (-52%), and "feels fearful or depressed" (-61%). Experiencing a selected sleep or stress disorder after four weeks among those who had the disorder at baseline was significantly more likely in those not physically active and/or not having lowered their BMI after four weeks. Changes in alcohol consumption and smoking did not significantly contribute to changes in the disorders. Those who failed to lower their coffee/tea use after four weeks were significantly more likely to have a sleep disorder and be easily emotionally upset. CONCLUSIONS: Changes in lifestyle behaviors after attending an educational program significantly reduced sleep and stress disorders in as little as four weeks, primarily explained by decreasing BMI and/or increasing exercise.

Phase transition affects energy transfer efficiency in phospholipid vesicles.

The fluorescence quenching of 6-propionyl-2-dimethylaminonaphtalene (PRODAN) and 6-dodecanoyl-2-dimethylaminonaphtalene (LAURDAN) by octadecyl rhodamine B (ORB) in a model system of small unilamellar vesicles (SUV) of dipalmitoylphosphatidyl-choline (DPPC) was investigated. Non-linear Stern-Volmer behaviour was observed in both systems in the gel phase (25 degrees C) and in the fluid phase (50 degrees C), resulting from association processes and from static quenching. The relative quenching efficiencies of both dyes depend on the phase state of the bilayer and indicate a deeper incorporation of PRODAN and LAURDAN into the membrane in its fluid phase than in its gel phase.

Coronary risk reduction through intensive community-based lifestyle intervention: the Coronary Health Improvement Project (CHIP) experience.

Vigorous cholesterol lowering with diet, drugs, or a combination has been shown to slow, arrest, or even reverse atherosclerosis. Residential lifestyle intervention programs have successfully lowered serum cholesterol levels and other coronary risk factors, but they have the disadvantages of high cost and difficulty with long-term adherence. Community-based risk-reduction programs have the potential to effect change at low cost and improve long-term adherence. To assess the effectiveness of, and to develop a model for, such programs, the community-based Coronary Health Improvement Project (CHIP) was developed in Kalamazoo, Michigan. In the intensive (30-day, 40-hour), hospital-based educational program, participants are encouraged to exercise 30 minutes a day and to embrace a largely unrefined plant-food-centered diet that is high in complex carbohydrates and fiber; very low in fat, animal protein, sugar, and salt; and virtually free of cholesterol. A total of 304 enrollees in the first program were at elevated risk of coronary artery and related diseases: 70% were > or =10% above their ideal weight, 14% had diabetes, 47% had hypertension, and 32% had a history of coronary artery disease. Of the enrollees, 288 "graduated" from the program (123 men, 165 women; mean age was 55+/-11 years). Various markers of disease risk, including serum blood lipids and fasting blood glucose concentrations, were measured before and after the program. At 4 weeks, overall improvements in the participants' laboratory test results, blood pressures, weights, and body mass indexes were highly significant (p <0.001). Triglyceride levels decreased significantly (p <0.05) in participants who had elevated triglyceride levels (>200 mg/dL in men, 200-299 mg/dL in women).

Effect of inhalation anaesthetics on the phase behaviour, permeability and order of phosphatidylcholine bilayers.

We have used differential scanning calorimetry and fluorescence anisotropy measurements to investigate the effect of five inhalation anaesthetics of diverse chemical structure (halothane, enflurane, n-pentane, chloroform and diethylether) on the phase behaviour of liposomes prepared from dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC), respectively. The incorporation of these anaesthetics induced a decrease of the phase transition temperature and/or a broadening of the phase transition peak depending on the transverse localisation of the investigated anaesthetic. At high anaesthetic concentrations we observed the disappearance of the pretransition peak and the appearance of a shoulder on the main phase transition peak due to the domain formation of the anaesthetics. An anaesthetic induced carboxyfluorescein efflux from the vesicle lumen was completed within a few minutes after the addition of the anaesthetics, probably resulting from a transient formation of membrane holes. All results are discussed with regard to the physicochemical properties of the anaesthetics applied.

Competitive carotenoid and cholesterol incorporation into liposomes: effects on membrane phase transition, fluidity, polarity and anisotropy.

Pure 1,2-dipalmitoyl-sn-glycero-3-phosphorylcholine (DPPC) or mixed DPPC:1,2-dipalmitoyl phosphatidyletanolamine (DPPE):1,2-dipalmitoyl diphosphatidylserine (DPPS) (17:5:3) liposomes were incorporated with 5 mol% dietary carotenoids (beta-carotene, lutein and zeaxanthin) or with cholesterol (16 and 48 mol%) in the absence or presence of 15 mol% carotenoids, respectively. The carotenoid incorporation yields ranged from 0.42 in pure to 0.72 in mixed phospholipid liposomes. They decreased significantly, from 3 to 14%, in the corresponding cholesterol-doped liposomes, respectively. Highest incorporation yields were achieved by zeaxanthin and lutein in phospholipid liposomes while in cholesterol-containing liposomes, lutein was highest incorporated. The effects on membrane structure and dynamics were determined by differential scanning calorimetry, steady-state fluorescence and anisotropy measurements. Polar carotenoids and cholesterol cause similar, dose-dependent effects: ordering and rigidification revealed by broadening of the transition peak, and increase of anisotropy. Membrane hydrophobicity is determined by cholesterol content and carotenoid polarity. In cholesterol-doped liposomes, beta-carotene is less incorporated than in cholesterol-free liposomes. Our observations suggest effects of carotenoids, even at much lower effective concentrations than cholesterol (8 to 80-fold), on membrane structure and dynamics. Although they are minor constituents of animal membranes, carotenoids may act as modulators of membrane phase transition, fluidity, polarity and permeability, and therefore, can influence the membrane physiology and pathology.

Heterogeneity of microsomal membrane fluidity: evaluation using intrinsic tryptophan energy transfer to pyrene probes.

Membrane fluidity measurements based on excimer formation of pyrene and pyrene derivatives as a measure of lateral diffusion yield a decreased fluidity in the presence of proteins [1-3]. It was the aim of our study to investigate whether the reduced excimer formation is due to a rigidifying effect of proteins on the whole membrane or if the fluorophore mobility is mainly hindered in the immediate protein environment. Resonance energy transfer in microsomal membranes between intrinsic tryptophan residues and pyrene were used to study the excimer formation rate in the vicinity of proteins. The excimer-to-monomer fluorescence ratio at excitation via resonance energy transfer is lower than that observed for the direct excitation. The results suggest that, because of a reduced fluidity in the neighbourhood of proteins, pyrene and pyrene fatty acids do not diffuse homogeneously in the membrane plane. A fluidity gradient exists from the membrane proteins to the bulk lipid.

Carotenoid incorporation into natural membranes from artificial carriers: liposomes and beta-cyclodextrins.

Liposomes and beta-cyclodextrin (beta-CD) have been used as carriers for the incorporation of three dietary carotenoids (beta-carotene (BC), lutein (LUT) and canthaxanthin (CTX)) into plasma, mitochondrial, microsomal and nuclear membrane fractions from pig liver cells or the retinal epithelial cell line D407. The uptake dynamics of the carotenoids from the carriers to the organelle membranes and their incorporation yield (IY) was followed by incubations at pH 7.4 for up to 3 h. The mean IYs saturated between 0.1 and 0.9 after 10-30 min of incubation, depending on membrane characteristics (cholesterol to phospholipid ratio) and carotenoid specificity. Mitochondrial membranes (more fluid) favour the incorporation of BC (non-polar), while plasma membranes (more rigid) facilitate the incorporation of lutein, the most polar carotenoid. A high susceptibility of BC to degradation in the microsomal suspension was observed by parallel incubations with/without 2,6-di-t-buthyl-p-cresol (BHT) as antioxidant additive. The beta-CD carrier showed to be more effective for the incorporation of lutein while BC was incorporated equally into natural membranes either from liposomes or from cyclodextrins. The presence of cytosol in the incubation mixture had no significant effects on the carotenoid incorporations.

Carotenoid incorporation into microsomes: yields, stability and membrane dynamics.

The carotenoids beta-carotene (BC), lycopene (LYC), lutein (LUT), zeaxanthin (ZEA), canthaxanthin (CTX) and astaxanthin (ASTA) have been incorporated into pig liver microsomes. Effective incorporation concentrations in the range of about 1-6 nmol/mg microsomal protein were obtained. A stability test at room temperature revealed that after 3 h BC and LYC had decayed totally whereas, gradually, CTX (46%), LUT (21%), ASTA (17%) and ZEA (5%) decayed. Biophysical parameters of the microsomal membrane were changed hardly by the incorporation of carotenoids. A small rigidification may occur. Membrane anisotropy seems to offer only a small tolerance for incorporation of carotenoids and seems to limit the achievable incorporation concentrations of the carotenoids into microsomes. Microsomes instead of liposomes should be preferred as a membrane model to study mutual effects of carotenoids and membrane dynamics.

Quantitative structure-activity relationship of coumarin derivatives. Involvement of partition between aqueous and membrane phase.

In order to understand the substrate behaviour of several 7-alkoxycoumarins and 7-alkoxy-4-alkylcoumarins towards the liver microsomal monooxygenase system, their lipophilic properties have been examined. As a model for the lipophilicity the reversed-phase liquid chromatographic retention parameter log kw has been used. In a system with methanol-water as the mobile phase and RP-18 (octadecylsilica) as the stationary phase, we found a quadratic relationship between the volume fraction of the organic solvent and the logarithm of the capacity factor (log k'). The extrapolation to a pure aqueous phase reveals a linear relationship of the theoretical capacity factor log kw with the chain length. This holds for 1-12 carbon atoms in the alkoxy chain and for zero to three carbon atoms in the alkyl chain. Moreover, the incremental effect of the methylene residues on the lipophilicity of the compounds (delta log kw/delta CH2) is found to be 0.60 +/- 0.01. If the coumarin derivatives are used as substates for the liver microsomal monooxygenase system, no systematic dependence of the enzymic data (Michaelis-Menten constant Km) on the lipophilic data (log kw) can be demonstrated. The metabolism of these compounds by the microsomal monooxygenase system seems not to be limited by the partition between the membrane and the aqueous phase. Whether other factors, e.g. the lateral diffusion of the substrates versus the membrane-bound enzyme system or enzyme active-site characteristics, govern the metabolism remains to be investigated.

Specific molecular properties of organic solvents determine the fluorescence depolarization of DPH and TMA-DPH in membranes.

In pig liver microsomes and protein-free egg PC liposomes the effects of organic solvent molecules on the fluorescence depolarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1-[4-(trimethylamino)phenyl]-6-phenyl-hexa-3,5-triene (TMA-DPH) were investigated. Aromaticity, alkyl chain length, and stereometry of the solvent molecules are shown to determine the changes of fluorescence depolarization. A concentration-dependent decrease in the fluorescence anisotropy is obtained with aromatic molecules but not with aliphatic molecules. The decrease correlates with the increasing side chain length of alkylbenzenes for both fluorophors and both membrane systems. Benzene in microsomes deviates characteristically from this trend. Measurements of TMA-DPH reveal smaller changes of anisotropy but yield the same qualitative results. Moreover, it is possible to distinguish the effects of the different stereometric properties of the three xylene isomers on the fluorescence anisotropy of DPH. According to their alkyl chain length and their specific stereometry, they exert the strongest fluidizing effect of all solvents investigated.

Pyrene excitation via resonance energy transfer from protein tryptophan reveals a fluidity gradient in liver microsomes.

Membrane fluidity measurements based on excimer formation of pyrene and pyrene derivatives as a measure of lateral diffusion yield a decreased fluidity in the presence of proteins [1,2]. It was the aim of our study to investigate whether the reduced excimer formation is due to a rigidifying effect of proteins on the whole membrane or if the fluorophore mobility is hindered mainly in the immediate protein environment. Resonance energy transfer in microsomal membranes between intrinsic tryptophan residues and pyrene was used to study the excimer formation rate in the vicinity of proteins. The excimer-to-monomer fluorescence ratio after excitation via resonance energy transfer decreased compared to that observed for the direct excitation. The results suggest that, because of the reduced fluidity in the neighborhood of proteins, pyrene and pyrenedodecanoic acid do not diffuse homogeneously in the membrane plane.

Radiation induced membrane changes and programmed cell death: possible interrelationships.

A short review of the evidence that lymphocyte membranes are a target for the initiation of irradiation induced programmed cell death (PCD) is given. It is assumed that for lymphocytes PCD represents an essential physiological mechanism in order to prevent degeneration of the biological system involved. Initiation of PCD can be obtained by a pharmacological activation as well as with irradiation. In both cases, protein kinase-C (PKC) is involved in the signal transduction from the cellular membrane to the nucleus where, by means of a metabolically active process, DNA fragmentation is induced. It is hypothesized that processes connected to lipid peroxidation in the cell membrane constitute a primary effect of irradiation induced PCD, where membrane fluidization or a compensatory process aimed to the maintenance of membrane fluidity (membrane homeoviscosity hypothesis) are likely to be involved.

Membrane fluidity of microsomal and thymocyte membranes after X-ray and UV irradiation.

A brief literature review shows that ionizing radiation in biological membranes and in pure lipid membranes causes malondialdehyde formation, indicating lipid peroxidation processes. With respect to membrane fluidization by ionizing radiation, in pure lipid membranes rigidization effects are always reported, whereas contradictory results exist for biological membranes. Starting from the assumption that membrane proteins at least partly compensate for radiation effects leading to a rigidization of membrane lipid regions, pig liver microsomes, as a representative protein-rich intracellular membrane system, were irradiated with X-rays or UV-C with doses up to 120 Gy at a dose rate of 0.67 Gy min-1 and up to 0.73 J cm-2 at an exposure rate of 16.2 mJ cm-2 min-1, respectively. For both irradiation types a weak but significant positive correlation between malondialdehyde formation and membrane fluidity is revealed throughout the applied dose ranges. We conclude that the membraneous protein lipid interface increases its fluidity under radiation conditions. Also, thymocyte ghosts showed an increased fluidity after X-ray irradiation. Fluidity measurements were performed by the pyrene excimer method.

Protein-dependent reduction of the pyrene excimer formation in membranes.

The presence of proteins in lipid bilayers always decreases the excimer formation rate of pyrene and pyrene lipid analogues in a way that is related to the protein-to-lipid ratio. Energy transfer measurements from intrinsic tryptophans to pyrene have shown (Engelke et al., 1994), that in microsomal membranes, the excimer formation rate of pyrene and pyrene fatty acids is heterogeneous within the membrane plane, because a lipid layer of reduced fluidity surrounds the microsomal proteins. This study investigates whether of not liposomes prepared from egg yolk phosphatidylcholine with incorporated gramicidin A give results comparable to those from microsomal membranes. The results indicate that the influence of proteins on the lipid bilayer cannot be described by one unique mechanism: Small proteins such as gramicidin A obviously reduce the excimer formation rate by occupying neighboring positions of the fluorescent probe and thus decrease the pyrene collision frequency homogeneously in the whole membrane plane, while larger proteins are surrounded by a lipid boundary layer of lower fluidity than the bulk lipid. The analysis of the time-resolved tryptophan fluorescence of gramicidin A incorporated liposomes reveals, that the tryptophan quenching by pyrene is stronger for tryptophans located closely below the phospholipid headgroup region because of the pyrene enrichment in this area of the lipid bilayer.

Fluidity of the microsomal membrane and cytochrome P450 reduction kinetics of pig liver microsomes as a consequence of organic solvent impact.

1. The effect of the aromatic solvents toluene, xylene and ethylbenzene on microsomal membrane fluidity and anaerobic NADPH-reduction kinetics were studied. 2. The relation of membrane fluidity to the kinetics of cytochrome P450 reduction by NADPH-cytochrome P450 reductase was examined with regard to a membrane-mediated molecular organization of the multienzyme components of the monooxygenase system. 3. Membrane fluidity changes were detected with the steady-state pyrene excimer formation method and with fluorescence lifetime measurements after incubation of the microsomes with organic solvents. 4. Increase in membrane fluidity in presence of organic solvents leads to a small but significant decrease of the rate constant of the cytochrome P450 reduction kinetics and a change in the relative amplitudes of the components of the biphasic response. 5. The results support the idea of a molecular organization of cytochrome P450 in clusters. Fluidization of the microsomal membrane by organic solvents increase the cytochrome P450 cluster formation.

Methyl-beta-cyclodextrins and liposomes as water-soluble carriers for cholesterol incorporation into membranes and its evaluation by a microenzymatic fluorescence assay and membrane fluidity-sensitive dyes.

A variety of methods to incorporate cholesterol into lipid membrane systems have been applied with varying success. We tested an incorporation method based on cholesterol-loaded methyl-beta-cyclodextrins and compared it to a method that uses cholesterol-loaded liposomes. With methyl-beta-cyclodextrin, we increased the cholesterol content in microsomal membranes to almost the fourfold of the original content. With cholesterol-loaded liposomes instead, we achieved an elevation of 140%. Short incubation times and well-defined carrier properties favor the beta-cyclodextrin method. For direct detection of membrane cholesterol, we slightly modified a microenzymatic fluorescence assay originally developed for precise cholesterol detection in serum. Without the need to perform lipid extraction, this assay was reliable for cholesterol detection in liposomes and in microsomes. Additionally, we compared the sensitivity of the fluidity-sensitive fluorescent dyes pyrene, pyrene-methanol, bis-pyrene, 1-6-phenyl-1,3,5,-hexatrien, and 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5,-hexatrien in order to detect cholesterol indirectly by the dynamically relevant changes exerted on lipid matrices. These dyes differ not only in their membrane location but also in their dynamical behavior. We calibrated the dyes in liposomes of defined cholesterol content and used the most suited ones to follow and quantify the cholesterol incorporation into liposomal and microsomal membranes.

Serum proteins of rats exposed to organic solvents examined by horizontal two-dimensional electrophoresis with an immobilized pH gradient in the first dimension.

Serum proteins of rats, exposed to methoxyethylacetate and a combination of methoxyethylacetate and isobutylacetate, were analyzed by horizontal high resolution two-dimensional gel electrophoresis with immobilized pH gradients in the first dimension. A total of 410 polypeptides were detected with either increasing or decreasing spot intensities after rat exposure to the organic solvents.

Calcium metabolism and osteoporotic ridge resorption: a protein connection.

The unique interrelationship between excess dietary protein, calcium metabolism, and osteoporosis with its associated ridge resorption has been reviewed. Recommendations for the prevention and management of osteoporosis have been discussed with concern for the calciuretic effect of a high protein diet customarily consumed in American society. Positive calcium balance promoted by the suggested treatment regimen may help to preserve ridge integrity and at the same time prevent the serious debilitating effects of generalized osteoporosis. Further research to evaluate for retardation and possible reversal of osteoporotic ridge resorption as affected by dietary protein intake is warranted.

Induction of apoptosis in thymocytes: new evidence for an interaction of PKC and PKA pathways.

The second messenger cascades connected to PKC and PKA are involved in the induction of apoptosis. Here we study the interaction of those two second messenger pathways with respect to the induction of apoptosis by stimulation or inhibition. The stimulators used were phorbol dibutyrate for PKC and one of the cAMP agonists Sp-5,6 DCl-cBIMPS or Sp-cAMP for PKA. The inhibitors used were staurosporin for PKC and the cAMP antagonist Rp-cAMPS for PKA. We found a synergism between both second messenger systems with regard to the induction of apoptosis in thymus lymphocytes.