PhD Dissertations by MUM Students – Physiology

Suppression of cytochrome P4501A (CYP1A1 and CYP1A2) mRNA levels in isolated hepatocytes by IL-1 and oxidative stress.

Order No. 9421867

Animals subjected to immunostimulatory conditions exhibit reduced tissue levels of total CYP and CYP dependent drug metabolism. We have investigated the possibility that depressed levels of two carcinogen-metabolizing CYP enzymes may be due to decreased levels of the mRNAs encoding these enzymes by studying the effect of monocyte-derived cytokines on the induction of CYP1A1 and CYP1A2 mRNAs in isolated rat hepatocytes. Medium conditioned by activated human peripheral blood monocytes or by the U937 monocyte cell line suppressed the induction of both mRNAs by TCDD, while beta-fibrinogen mRNA levels increased 30- to 40-fold. Recombinant interleukin-1 suppressed the inducer-dependent accumulation of both CYP1A1 and CYP1A2 mRNAs in a dose-dependent fashion, while two other monocyte derived cytokines, interleukin-6 and transforming growth factor-beta, did not. Run-on transcription analysis demonstrated that conditioned medium and interleukin-1 rapidly suppressed the transcription rate of CYP1A1 and CYP1A2 in inducer-treated hepatocytes. Since many of the actions of inflammatory mediators can be mimicked by oxidative stress, we also treated isolated hepatocytes with various concentrations of H(sub 2) O(sub 2) (0.25 to 1.0 mM) to investigate the possibility that expression of these genes may also be modulated by oxidative stress. Inducer-dependent accumulation of CYP1A1 and CYP1A2 mRNAs were maximally reduced approximately 50 and 70% respectively by 1.0 mM H(sub 2) O(sub 2). Run on transcription analysis suggested that the effect of H(sub 2) O(sub 2) on these mRNAs was mediated transcriptionally. The reduction in CYP1A mRNA levels was not due to a reduction in the levels of all mRNAs due to some general toxic effect since glyceraldehyde-3-phosphate dehydrogenase, alpha-tubulin, beta- fibrinogen and albumin mRNA levels did not change or were actually increased, and lactate dehydrogenase released into the medium was not increased, with H(sub 2) O(sub 2) treatment. Insulin also reduced the expression of both mRNAs, and N- acetylcysteine, which increases intracellular glutathione levels, completely reversed the insulin effect on both mRNAs and the H(sub 2) O(sub 2) effect on CYP1A1 mRNA but only partially reversed the H(sub 2) O(sub 2) effect on CYP1A2 mRNA. Source: DAI, 55, no. 04B, (1994): 1294

Reduced cardiovascular and all-cause mortality in older African Americans practicing the Transcendental Meditation Program

Order No. 9701126

African Americans have a well-documented excess of CVD mortality which is at least in part due to psychosocial stress. The Transcendental Meditation® (TM) program has been reported to reduce psychological stress, cardiovascular risk factors and incidence of heart disease. A randomized controlled trial indicated that TM reduced hypertension significantly more than progressive muscle relaxation (PMR) and an educational control (EC) in older (mean age = 67 years) Africans Americans after 3 months. Pilot research in Caucasian elderly has found a 73% reduction in all-cause and cardiovascular (CVD) mortality in the TM group compared to the combined control group.

Based on these findings, TM (n = 36) was hypothesized to reduce incidence of all-cause and CVD mortality compared to PMR (n = 37) and EC (n = 36) and a combined control (CC, n = 73) group among the African American participants with mild hypertension in the original BP study. After 5 years, an all-cause and CVD mortality follow-up was conducted with data provided from Vital Statistics, Sacramento, CA. Survival distributions were compared by the Wilcoxon and Cox proportional hazards tests. There were 0.0% (0/36) CVD fatalities for TM compared to 9.5% (7/73) for CC, and 8.5% (3/36) all-cause fatalities for TM compared to 19% (14/13) for CC. Both all-cause (P =.045) and CVD (P =.021) mortality were significantly lower for TM compared to combined controls. The relative risk (RR) for TM compared with combined controls was 0.00 (95% CI 0-0.63) for CVD mortality and 0.32 (95% CI 0-0.96) for all-cause mortality.

These findings suggest that TM practice may reduce incidence of CVD and all-cause mortality in older hypertensive African Americans. According to Maharishi’s Vedic Approach to Health, TM enhances the holistic inner intelligence of mind and body, and thereby promotes balance in psychophysiological functioning and thus helps prevent premature disease and death. The demonstrated benefit for the Transcendental MeditationÒ program seems to have important implications for clinical and public health policy for reducing excessive CVD and all-cause mortality in African Americans. Source: DAI, 57, no. 08B, (1996): 4999

Effects of a multimodal approach of Maharishi consciousness-based health care on carotid atherosclerosis: A study of coronary artery disease patients

Order No. 3131257

Cardiovascular disease (CVD) is still the largest contributor to morbidity and mortality in the world. Over the past 30 years, focus on primary and secondary prevention of cardiovascular disease and its related risk factors have yielded three major prevention strategies-drug therapies, lifestyle modification and stress reduction therapies. Despite this effort, up to 50% of patients with documented CVD have recurrent cardiac events.

A new angle that supports further prevention of CVD is needed. Maharishi Consciousness-Based Health Care system, a natural, prevention-oriented system of health, includes 40 modalities for enlivening the “inner intelligence of the body,” which are responsible for coordinating diverse physiological systems into an integrated whole.

This pilot trial compared effects of four Maharishi Consciousness-Based Health Care modalities to those of usual care on carotid intima-media thickness (IMT) in elderly subjects with documented cardiovascular disease and two to six CVD risk factors. The Maharishi Consciousness-Based Health Care modalities included the Transcendental Meditation program, neuro-physiological integration exercises, dietary and herbal supplement approaches. Usual care included the secondary prevention system offered at the University of Iowa Hospitals and Clinics, which are based on practice guidelines promoted by the American Heart Association.

Twenty-eight volunteer subjects were matched on age (mean 72 years), gender and severity of documented CVD. Measures were taken for baseline and nine-month posttest. At baseline the experimental group had significantly higher BMI (30 versus 26), triglycerides (177 mg/dl versus 101 mg/dl, and blood pressure (137 versus 120 mm Hg). They were more often single, and had lower income. Covarying for these baseline differences in major CVD risk factors, the experimental group tended to show a greater decrease in mean common carotid IMT after nine months. (Experimental -0.023 mm, Usual Care +0.041 mm, p = 0.07). The IMT regression in the experimental subjects was associated with high compliance. There was a strong correlation between compliance and increases in physical, mental and behavioral strength (r = 0.47), as assessed by Maharishi Consciousness-Based Health Care procedures. These findings suggest that enlivening the body’s inner intelligence could be an effective tool to deal with the current epidemic of cardiovascular disease.

Complex dynamics in biological systems: Spontaneous variations of serotonin uptake into platelets as a model of signal control in the central nervous system

Order No. 9228948

Coherent fluctuations in the level of enzymatic activity in solutions of purified enzymes have been demonstrated in several laboratories. The existence of such fluctuations involving simple enzymes in solution suggests that coordinated behavior among macromolecules can occur in the absence of synchronizing cues from the environment. The present research sought to determine whether spontaneous fluctuations in the frequency range of 0.01 to 0.25 cycles/minute occur in the initial rate of uptake of serotonin (also known as 5-hydroxytryptamine, 5-HT) into platelets, a protein-mediated function. Such fluctuations were found, and their temperature dependence and response to partial inhibition by imipramine were examined. The level of uptake was determined as the amount of ($\sp3$H) 5-HT taken up by platelets in a one-minute incubation at 37$\sp\circ$C. Spectral analysis provided a measure of the frequency content of each time series of initial rates. The average behavior of many time series was displayed by summing their individual spectra. Statistical significance of individual frequency estimates was determined by Fisher’s test or Siegel’s test. The major findings of this research were: (1) apparently spontaneous periodic and aperiodic oscillations in the initial rate of 5-HT uptake, (2) an apparent shift to faster frequencies with an increase in sample storage temperature, (3) alterations in the frequency or phase of fluctuations in the presence of imipramine, (4) absence of an effect of a brief pre-sonication of platelet preparations on 5-HT uptake patterns. The findings suggest that coherent variations in 5-HT uptake may be organized by factors intrinsic to the platelet suspensions. Fluctuations in 5-HT uptake rate could be due to oscillatory variations of metabolic parameters in the platelet or to spontaneous conformational fluctuations in the uptake protein. The mechanism mediating ordering effects between platelets could involve alterations in water structure associated with such configurational changes or possibly alterations of diffusible chemical species such as intermediates of metabolism or metal ions. Since the 5-HT uptake system of neurons is essentially identical to that of platelets, these findings may help to explain serotonin-mediated rhythmicities in signal transmission in brain areas rich in non-classical serotonergic nerve endings.

Lifestyle Modifications and Healthy Biological Aging: Effects of Telomerase Activity and Telomere Length.

Order No. 3523287

Lifestyle modifications such as practicing the Transcendental Meditation (TM) program, maintaining a healthy diet and regular exercise could help to overcome age-related disorders and promote healthy biological aging. Biochemical, physiological, and psychological studies have revealed their positive effect, although the effect on telomerase gene expression and telomere length is poorly understood. Therefore, the goal of this research study is to determine whether lifestyle modifications will support healthy biological aging. Two independent studies were conducted for this purpose. First, a National Institute of Health (NIH) funded randomized pilot study was conducted using two intervention groups (24 TM + health education (HE) group and 24 enhanced health education (EHE) group) with Stage I hypertension African Americans.Second, a cross-sectional study was conducted using 19 long-term meditators and 19 nonmeditator controls. Telomerase gene expression (hTR and hTERT) and relative telomere length of peripheral blood cells by quantitative real-time PCR technique was measured in both studies.

In the NIH pilot study, Wilcoxon matched pairs tests showed a significant difference in the medians for hTERT (TM + HE = 0.03, p = 0.05; EHE = 0.60, p < 0.01) and hTR (TM + HE = 0.34, p < 0.001; EHE = 5.48, p < 0.001) in both groups. Dependent t-tests showed significant differences for systolic BP (TM + HE = -5.53 ± 11.23, p = 0.02; EHE = -9.00 ± 11.41, p < 0.001) in both groups and diastolic BP in the EHE group (EHE = -4.93 ± 7.05, p < 0.01).These findings suggest that intensive lifestyle modifications may be effective in promoting healthy biological aging.

In the cross-sectional study, the Kruskal-Wallis test indicated a significant trend for hTR in long-term meditators (long-term meditators = 0.74; non-meditator controls = 0.45, p = 0.08). Analysis of covariance showed a significant difference in psychological stress in long-term meditators (long-term meditators = 26.60 ± 3.95 non-meditator controls = 43.20 ± 0.68,p < 0.001), adjusting for BMI, exercise, smoking, intake of vitamins and omega-3. Future research is warranted with larger sample sizes to further evaluate the impact of TM on telomerase gene expression and telomere length.

Analysis of CYP1A1 gene chromatin structure—evidence for multiple translationally positioned nucleosomes.

Order No. 9722238

In vivo low-resolution indirect end-labeling analysis of CYP1A1 gene chromatin structure revealed precisely positioned nucleosomes in the enhancer, promoter and transcribed regions. In vivo high-resolution LMPCR analysis of the CYP1A1 regulatory region revealed multiple micrococcal nuclease (MNase) cleavages with different relative intensities suggesting that multiple translationally positioned nucleosomes occupy the CYP1A1 regulatory region in major and minor translational nucleosome frames. In both low-resolution and high-resolution experiments, positions of nucleosomes did not change when CYP1A1 gene transcription was induced with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). In clone-11 cells, TCDD-induction globally increased the sensitivity of the CYP1A1 regulatory region to MNase indicating induced alterations in chromatin structure. In vitro nucleosome reconstitution and hydroxyl radical footprinting of the CYP1A1 enhancer demonstrated the presence of multiple overlapping translationally and rotationally positioned nucleosomes. In vivo footprinting and high-resolution LMPCR analysis of the CYP1A1 enhancer region in conjunction with the in vitro electrophoretic mobility shift analyses demonstrated that, following the treatment of cells with TCDD, the kinetics and time course of in vivo interactions at xenobiotic response elements (XREs) and at diverse sequence xenobiotic responsive elements (DXEs), and the kinetics and time course of DNA binding activity of aryl hydrocarbon receptor (AHR) paralleled the kinetics and time course of increased MNase accessibility of the CYP1A1 enhancer region in clone-11 cells, but not in clone-3 cells. Even though nuclear extracts prepared from both cell lines contained AHR that was capable of binding to XRE1, TCDD treatment results in the appearance of in vivo footprints at XREs and DXEs in the clone-11 cell line, but not in clone-3 cell line indicating the presence of an in vivo inhibitory activity that prevents the binding of AHR to XREs in the clone-3 cell line. The results of experiments presented in this thesis indicated that ligand-dependent and AHR-dependent chromatin alterations occur in the regulatory region of CYP1A1 gene without an accompanying change in the multiple translational positions of nucleosomes, and support the hypothesis that such alterations may be one transcriptional regulatory mechanism contributing to the high expression of CYP1A1 gene in clone-11 cells.

Systematic Reviews and Meta-Analyses on Effects of Ayurvedic Interventions for Hypercholesterolemia, Hypertension, and Coronary Heart Disease

Order No. 10617601

Heart disease is the number one cause of death globally. Because of numerous side effects and increasing cost of conventional treatments, there is a growing interest in complementary healing approaches like Ayurveda. However, in lack of sufficient scientific evidence, safety and efficacy profile of these interventions has not yet been established. Systematic reviews and meta-analysis are the gold standard of evidence upon which clinicians, consumers, policymakers rely. Until this time, there are no Cochrane or other systematic reviews and meta-analysis on Ayurvedic interventions for cardiovascular disorders. This study was conducted with an aim to explore the efficacy of Ayurvedic interventions in hypercholesterolemia, hypertension and coronary heart disease, strength of the evidence and their possible side effects.

Cochrane guidelines for systematic reviews and meta-analysis were followed to design, formulate and implement search strategies, select studies, collect, abstract and analyze data, assess risk of bias, and report and interpret results.

Evidence from results were classified as per guidelines from American Academy of Neurology. Three meta-analyses of 64 studies on 2629 people studying effects of 10 different Ayurvedic interventions for hypercholesterolemia, high blood pressure and coronary heart disease, concluded that there is moderate to high strength evidence that several Ayurvedic herbal preparations are safe and effective. They pose no known side effects and thus can be used as dietary supplements or as an adjuvant to conventional therapy for better results. It was observed that Commiphora mukul (guggulu) reduced total cholesterol and low density lipoprotein levels by approximately 16 mg/dL and 18 mg/dL respectively with high certainty evidence. Similarly, garlic and Terminalia arjuna (arjuna) based formulas also had high to moderate strength evidence of their efficacy to reduce cholesterol levels. On the other hand, Arjun Vachyadi compound and Rauwolfia serpentina based formulas were found to have moderate certainty evidence to reduce high blood pressure. It was also observed that Ayurvedic formulas with arjuna as a chief ingredient are capable of improving left ventricular ejection fraction by 12 % with a moderate strength of evidence. Findings of these systematic reviews and meta-analysis encourage future researchers to conduct methodologically rigorous randomized clinical trials studies with a larger sample size.

Effects of the “Transcendental Meditation” (rtm) program on neuroendocrine indicators of chronic stress

Order No. 9806955

Reduction of stress and its effects is an important objective because evidence suggests that stress causes or aggravates almost every human disease. The Transcendental Meditation$\sp\circler$ (TMS) program is a widely studied stress-reduction approach; thus, understanding the neuroendocrine mechanisms mediating its effects would be useful. A previous cross-sectional study comparing long-term practitioners of the TM program to matched controls showed highly significant differences in urinary variables reflecting neuroendocrine function. Practitioners of the TM program had increased urinary excretion of dehydroepiandrosterone and nighttime 5-hydroxyindoleacetic acid (5-HIAA, the major metabolite of serotonin), decreased excretion of cortisol, aldosterone, the norepinephrine/epinephrine metabolite vanillylmandelic acid, zinc, calcium and sodium, and lower scores on tests of anxiety and mood disturbance. These and other results suggested that the differences were due to reversal of the long-term effects of stress by the TM program. The current prospective, random-assignment study attempted to test this hypothesis. Healthy, Caucasian men (18-32 y) were randomly assigned to either the TM program or a stress education control (SEC). Before and after four months’ practice of the assigned stress management program, three consecutive 8-hour urine collections were taken, and psychological self-report tests were administered. Urine samples were analyzed for 5-HIAA by spectrophotometry, for adrenocortical steroids by radioimmunoassay and for ions by atomic absorption spectrometry. Compared to controls, TM subjects showed a significant decrease in sodium excretion during the afternoon-evening period, near-significant decreases in calcium excretion during the afternoon-evening and 24-hour periods, and statistically insignificant decreases in excretion of the other three ions. High-compliance subjects from both groups showed significantly lower cortisol excretion over the 24-hour period than low-compliance subjects, suggesting that high compliance with either program leads to a reduction in cortisol. Results for sleeptime 5-HIAA were in the predicted direction, though not significant. Increased regularity of practice of the TM program was associated with a decreased POMS Tension-Anxiety score. Some meaningful changes may not have reached significance due to inadequate statistical power. Results were generally consistent with previous findings.

Mapping of sequence specific DNA- protein interactions: a versatile, quantitative method and its applications to transcription factor XF1.

Order No.9608533

Mapping the consensus sequence of DNA binding proteins has been greatly accelerated by methods that use in vitro selection of high affinity sequences from a library of random DNA molecules, followed by PCR-amplification and sequence analysis. However, these methods lose other valuable information because they use repetitive cycles of selection and amplification.

We have developed a method that overcomes this limitation, not only defining the consensus sequence, but also quantitating the effect on DNA-protein affinity of replacing each base in the recognition domain with every other base. The features of this method are: (1) Instead of synthesizing one oligonucleotide population containing a long randomized domain, we synthesize several oligonucleotide populations, each randomized at two positions. Because only a few species are present in each population, the concentration of each is sufficient to saturate the DNA-binding protein. Consequently, the abundance of each protein-bound oligonucleotide accurately reflects its binding affinity. (2) Because only a few species are represented in each oligonucleotide population, a single round of selection and amplification generates sufficient material for sequencing. This avoids biasing the population of protein-bound oligonucleotides toward high affinity species. Consequently, the abundances of oligonucleotides determined by sequence analysis accurately reflects their binding affinities. (3) We developed data collection and analysis procedures that eliminate artifacts, and yield accurate measures of: (a) the selectivity of the protein for each base at each position within the recognition domain (normalized relative selectivity), (b) the contributions of individual sites within the recognition domain to the binding affinity (selectivity variance), (c) the relative affinity of a particular sequence for the DNA-binding protein (global selectivity). (4) We developed a procedure for deducing aspects of the matrix of hydrogen bonds involved in DNA-protein interactions.

This method was first developed and applied to the nuclear protein XF1, which binds to xenobiotic responsive elements, that class of elements through which the liganded Ah receptor activates transcription of the CYP1A1 gene. We confirmed results by (1) cloning and sequencing individual XF1-bound oligonucleotides, and (2) competition EMSA analysis of oligonucleotides designed on the basis of in vitro selection results. Source: DAI, 56, no. 11B, (1996): 5943

Mechanisms relating stress reduction and health: changes in neuroendocrine responses to laboratory stress after four months of Transcendental Meditation

Order No. 9534651

Pharmacological treatments have as yet failed to show clear reduction in the risk of development of coronary heart disease (CHD). As a result, behavioral treatments such as stress reduction programs continue to receive attention as alternative approaches for prevention as well as for treatment of heart disease. Research on the Transcendental Meditation (TM) technique of Maharishi Mahesh Yogi has shown it to be effective in reducing hypertension and also responsible for decreased basal cortisol levels, both acutely with the practice and longitudinally. In this study, the longitudinal effects of TM and a stress education control (SEC) on neuroendocrine responses to acute laboratory stressors were investigated.

The purpose of the present research was to examine in healthy male caucasians (18-32 yrs) the acute effects of laboratory stressors on plasma cortisol, serotonin, catecholamines, thyroid-stimulating hormone (TSH), growth hormone (GH), testosterone and dehydroepiandrosterone (DHAS) during the stress session, and changes in their responses to stress after four months’ participation in either stress management approach.

Plasma for cortisol, serotonin and the catecholamines was sampled periodically throughout the one-hour stress session using a continuous blood withdrawal pump, whereas samples of GH, TSH, DHAS and testosterone were sampled for 4 min at the beginning and at the end of the session. The laboratory stress session consisted of mental arithmetic (6 min), a mirror star tracing task (3.5 min), and isometric hand grip (3.5 min), separated by 25 min rest periods. Samples for cortisol, GH, TSH and testosterone were assayed by radioimmunoassay and statistically analyzed by t-test and one-way repeated measures ANOVA.

When compared to the SEC group by ANCOVA, basal cortisol levels and the average cortisol levels across the stress session decreased, while cortisol responsiveness increased, for the TM group after four months’ practice. For the TM group, TSH response to stress decreased while GH and testosterone responses increased over the same period. Plasma serotonin baseline, average and response to stress during the session showed a rise for the SEC group and a fall for the TM group over four months of intervention. No differences between the two groups in the changes in catecholamine responses to stress from pre- to posttest were noted, likely due to the small sample size.

These results indicate that practice of the Transcendental Meditation technique is associated with lowered plasma serotonin and cortisol as well as increased cortisol response to acute stress, in addition to changes in the responses of GH, TSH and testosterone to acute stressors. It is suggested that not only the changes in cortisol but also changes in basal level or response of other hormones reflect reduction of, or resistance to, the effects of chronic stress, i.e., changes towards more optimal adaptive mechanisms. (Abstract shortened by UMI.) Source: DAI, 56, no. 06B, (1995): 3074

Effect of the Transcendental Meditation Program and Health Education on Allostatic Load: Promoting Normal Aging

Order No. 3475629

The “weathering” hypothesis of aging suggests that African Americans experience accelerated aging due to the cumulative effects of stress, which causes multisystem “wear and tear” or allostatic load. Previous research shows that lifestyle changes can improve individual biomarkers of allostatic load, leading toward greater health and normal aging. Specifically, practice of the Transcendental Meditation (TM) program and changes in diet and physical activity reduce biological aging factors. This exploratory study examines the combined effects of the Transcendental Meditation program and conventional health education of diet and physical activity on allostatic load in 19 African American women and men with stage I hypertension over a 4-month period. This is a sub-study to a parent study on hypertension mechanisms conducted at Howard University Medical Center in Washington, DC.

The primary outcome of this study was a seven biomarker allostatic load index: body mass index (BMI), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), systolic blood pressure (SBP), diastolic blood pressure (DBP), glycosylated hemoglobin (HbA1c), and dehydroepiandrosterone sulfate (DHEAS). Each of the individual biomarkers of the allostatic load index and a psychosocial stress measure, the General Health Questionnaire (GHQ-20), were also analyzed.

Results showed a significant reduction in the allostatic load index based on composite T scores (pretest mean = 22.73 ± 2.83 and posttest mean = 20.13 ± 4.66; t (18) =3.46; p=0.003; effect size=0.92), and a marginally significant reduction in the allostatic load index threshold scores (2.63 ± 1.01 vs. 2.05 ± 1.13; t (18) =1.93; p=0.069; effect size=0.57). Results showed a significant reduction in the individual biomarkers of SBP (144.9 ± 7.29 vs. 136.72 ± 12.51; t (18)=2.94; p=0.009; effect size=1.12) and DBP (86.16 ± 5.44 vs. 81.30 ± 8.5; t (18)=2.83; p=0.012; effect size=0.89), and in the psychological distress measure: GHQ (23.54 ± 11.96 vs. 15.42 ± 10.34; t (18)=3.75; p=0.001; effect size=0.68). Other individual biomarkers also changed in the predicted direction, but the changes were not statistically significant.

Findings suggest that changing lifestyle with the Transcendental Meditation program and health education of diet and physical activity may reduce overall allostatic load in hypertensive African Americans.

Prevention of cardiovascular disease in Maharishi Ayur-Veda participants: a cross-sectional study of carotid atherosclerosis

Order No. 3374437

Cardiovascular disease (CVD) is the number one cause of mortality in developed countries. Previous research on Maharishi Ayur-Veda indicates reduction in cardiovascular risk factors and events, including decreased blood pressure, carotid atherosclerosis and all cause mortality.

Maharishi Ayur-Veda is Maharishi Mahesh Yogi’s revival of an ancient system of natural health care and includes mind, body, behavioral and environmental modalities to enliven the field of consciousness at the basis of all physiological functioning.

The purpose of this cross-sectional study was to investigate effects of long-term practice of Maharishi Ayur-Veda in a community setting on carotid artery blockage, blood pressure and serum lipid measures. Possible mediators of carotid blockage were also explored.

One hundred fifty-four adult subjects (n=74 MAV and n=80 controls) were included in this study. The Maharishi Ayur-Veda group included subjects from southeast Iowa who had been practicing the Transcendental Meditation (TM) program for greater than 5 years (mean=27 years). Among other MAV modalities widely practiced were the group practice of the TM-Sidhi program with Yogic Flying, yoga postures (asanas), a breathing exercise (pranayama), herbal supplements, and vegetarian diet. Controls were selected from the Stroke Detection Plus database of southeast Iowa clients.
All subjects were measured on the primary outcome, carotid artery blockage, by modified duplex ultrasound.

Analysis of covariance (ANCOVA), controlling for age, gender, body-mass index, family history of cardiovascular disease, smoking and exercise indicated significantly less carotid blockage in the MAV group (0.26 ± .37) compared to controls (0.36 ± .62) (p = 0.01). ANCOVA indicated significant difference between MAV and controls on systolic blood pressure (MAV 118.45 mm Hg ± 13.37, vs. controls 129.70 mm Hg ± 15.52; p=0.01), and HDL cholesterol (MAV 60.60 mg/dL ± 15.09, vs. controls 47.78 mg/dL ± 13.96; p=0.05). Systolic blood pressure was found to be a possible mediator of the effects of treatment on carotid blockage.

The results of this study provide further evidence for the effect of Maharishi Ayur-Veda on CVD risk factors. It is recommended that Maharishi Ayur-Veda, as a comprehensive preventative approach to cardiovascular disease, be incorporated into public health programs.

Mechanisms of AH receptor down-regulation: Involvement of a labile protein, a calcium dependent protease, and a protein kinase

Order No. 9633803

In this dissertation we present evidence that CYP1A1 transcription and in vivo DNA-protein interactions at XREs are down-regulated in parallel with the DNA-binding activity of the ligand activated AH receptor complex (AHRC). This indicates that down-regulation of AH receptor DNA-binding activity is important in regulating CYP1A1 transcription, and that the AHRC is required continuously to maintain transcription. We show also that the down-regulation process depends on protein synthesis, and that it involves degradation of the AHR subunit but not of ARNT. AHRC down-regulation is a Ca$sp{2+}$ dependent process, since both depletion of intracellular Ca$sp{2+}$ stores, and interference with Ca$sp{2+}$ currents can inhibit down-regulation in Hepa-1 cells. We also demonstrate that a specific inhibitor of the Ca$sp{2+}$ dependent protease calpain, as well as the protein kinase inhibitors H-7, calphostin C, and bisindolylmaleimide can block AHRC down-regulation. These findings are functionally relevant, since treatments that block down-regulation increase in AHRC dependent CAT gene expression substantially. Ca$sp{2+}$ measurements reveal a very rapid and transient change in intracellular free (Ca$sp{2+}$) in response to 2,3,7,8- tetrachlorodibenzo- (p) -dioxin (TCDD).

Thus, TCDD orchestrates a second response, that starts with a rapid rise in (Ca$rmsp{2+}rbracksb{i}$ and results in the activation of a Ca$sp{2+}$ dependent protease, which in turn is instrumental in AHRC down-regulation. Down-regulation involves components in addition to calpain since: (1) In vitro calpain digestion of the AHRC results in partial digestion products not observed in vivo, which implicates secondary proteases. (2) Calpains are too stable to mediate cycloheximide action. (3) Down-regulation can be blocked by cycloheximide after TCDD-induced $rmlbrack Casp{2+}rbracksb{i}$ transients have passed.

Finally, we present data suggesting that ionomycin might induce AHRC/ARNT complex formation in a ligand independent manner. Also, experiments with caffeine show that AHRC dependent CAT gene expression can be elevated without changing the levels of liganded AHRC, the kinetics of AHRC activation, or down- regulation. This increase in AHRC mediated transactivation is probably due to elevated $rmlbrack Casp{2+}rbracksb{i}.$ Source: DAI, 57, no. 06B (1996): p. 3580

Mechanisms of inflammation in ulcerative colitis: a role for neutrophils and their free radicals

Order No. 9836285

Ulcerative Colitis (UC), a common form of Inflammatory Bowel Disease (IBD), is characterized by recurrent episodes of acute colonic inflammation, associated with abdominal pain, cramping, and bloody diarrhea. Since current treatments for IBD are not ideal, there is a need to develop better clinical management of UC. We chose to investigate pathophysiological mechanisms through which inflammation and tissue damage in UC occurs.

Hypothesis. (1) The final common pathway which leads to tissue damage in UC is mediated primarily by reactive oxygen species (ROS), (2) neutrophils are the main source of these ROS, and (3) these neutrophils are attracted to the colonic mucosa and activated by circulating (plasma) factors and local (colonic) factors.

Aims and methods. To support the above hypothesis, we proposed three aims: (Aim 1) To determine whether plasma from UC patients is pro-inflammatory. To this end, we evaluated the respiratory burst of PMN after incubation with plasma from UC patients. (Aim 2) To determine whether colonic factors in UC patients are pro-inflammatory. To this end, we evaluated the expression of the PMN adhesion molecule CD11b after stimulation with colonic factors from UC patients. (Aim 3) To determine whether colonic tissues from UC patients have abnormally high levels of oxidative products. To this end, we developed and used a novel immunoblotting technique to analyze oxidation products in colonic tissue from IBD patients. Colonic tissues were analyzed for protein carbonyls, nitrotyrosine, and 4- hydroxynonenal (4-HNE).

Results and discussion. (1) Plasma from UC patients significantly enhanced the PMN oxidative burst compared to plasma from controls. (2) Colonic factors from patients with UC significantly up-regulated CD11b compared to colonic factors from controls. These two results suggest that plasma and colonic factors in UC are pro-inflammatory, and may, therefore, perpetuate chronic inflammation. (3) Nitrotyrosine and 4-HNE were significantly higher in CD than in controls. In UC, nitrotyrosine and 4-HNE were also elevated, but these values did not reach significance. These results suggest that the ulcerations and tissue damage, which are hallmark features of IBD, may be the result of above normal oxidative stress. There were no differences between the groups for protein carbonyls. Source: DAI, 59, no. 06B, (1998): 2698

Functional role of diverse sequence xenobiotic response elements (dxes) in regulation of cytochrome P-4501a1 (CYP1A1) gene transcription

Order No. 9636934

This dissertation presents evidence that activation of CYP1A1 gene transcription by aryl hydrocarbons is a multicomponent process involving interactions of the AH receptor complex (AHRC) with xenobiotic response elements (XREs) and interactions of secondary transcription factors with diverse sequence xenobiotic response elements (DXEs) and that interactions at DXEs are functionally important and are dependent on interactions of functional AHRC with XREs. Interactions at DXEs appear to be important due to the fact that DXEs can cooperate with XREs to confer on a reporter gene responsiveness to aryl hydrocarbons and the fact that the appearance of in vivo interactions at DXEs during activation parallels those at XREs as well as the fact that the kinetics of appearance of these interactions correlate with the levels of expression of P4501A1 mRNA in different cell lines. Dependence of interactions at DXEs on the interactions of functional AHRC with XREs is supported by the following findings: (1) Interactions at XREs and DXEs appear in parallel. (2) Down-regulation of the AH receptor leads to disappearance of footprints at XREs and DXEs. (3) Inhibition of protein synthesis which is known to prevent down-regulation of the AH receptor, preserves footprints at DXEs and XREs. (4) Cell line- specific differences in the kinetics of in vivo interactions at XREs are parallel at DXEs. (5) AHs fail to induce in vivo footprints at both XREs and DXEs in Hepa 1 mutant cells lacking functional nuclear AH receptor complex. (6) The appearance and disappearance of in vivo footprints at DXEs could not be correlated with changes in constitutive DXE-specific DNA- binding activities and instead correlated with changes in the DNA-binding activity of the AH receptor. The in vivo and in vitro findings reported here regarding the relationship between the interactions at XREs and DXEs are consistent with a chromatin remodelling mechanism during activation of CYP1A1 transcription which is induced by the activated AH receptor exposing previously inaccessible DXEs for interactions with constitutively present nuclear factors. Finally, a link is made between CYP1A1 gene regulation, the unified field described by physics and Maharishi’s Vedic Science. Source: DAI, 57, no. 07B, (1996): 4210

Transcendental Meditation technique: a new direction for smoking cessation programs.

Order No. 9000436

This prospective cohort study examined the effect of practice of the TM technique on smoking behavior during a period of 20 months. The subjects were 7070 individuals over 16 years of age who attended introductory lectures on the TM technique. Nine hundred and twenty-five (13%) of these learned the TM technique and 6145 (87%) did not. Prior to attending the lecture there were no differences in demographic variables or in smoking habits between the TM and non-TM groups. At the end of the study however, 33% of the smokers in the TM group had quit smoking as compared to 21% of those in the non-TM group (df = 1, chi squared = 3.85, p <.05). When regularity of practice of the TM technique was taken into account, it was found that 60% of those who meditated regularly twice each day had quit smoking as compared to 41% for those who meditated once each day, 21% for those who meditated irregularly or had stopped the practice, and 21% for those who never learned the TM technique (df = 3, chi squared = 18.25, p =.0004). The quit rates represented on the average a 12-month cessation period (SD = 7.57) for the meditators and a 10-month cessation period (SD = 8.54) for the non-meditators.

Furthermore, when quit and decreased rates were combined, it was found that 90% of those who practiced TM twice each day had quit or decreased smoking by the end of the study vs 71% for the once each day TM meditators, 55% for those who were irregular or no longer practiced TM, and 33% for the non-TM group (df = 3, chi squared = 35.734, p <.0001). These results strongly suggest a correlation between frequency of practice of the TM technique and increased likelihood of stopping smoking. Source: DAI, 50, no. 08B, (1989): 3428

Extensive artifact research was conducted. Some of the weighings were done under constant temperature and constant relative humidity.

Significant deviations from constancy of weight were found in the range of 0.2 to 1.5 mg for the 250-ml flasks and up to 0.06 mg for the 25-ml flasks. For sprouting seeds deviations of the same magnitude were found. Deviations of positive and negative amounts and zero results were obtained. Results were reproduced in blind experiments by five different experimenters. Experiments during eclipses implied a relationship of weight changes with planetary constellations. Source: DAI, 54, no. 03B, (1993): 1449

Selective growth inhibition of human colon adenocarcinoma and malignant melanoma cell lines by sesame oil in vitro

Order No. 9133557

Ayurveda, an ancient, comprehensive and prevention-oriented system of natural health care, has highly recommended the topical use of sesame oil above other oils. Applied to the skin on a daily basis and to the colon on a seasonal basis, it is claimed to improve physiological balance, vitality and longevity. Sesame oil contains relatively high levels of the essential polyunsaturated fatty acid, linoleic acid, in the form of triglycerides. The antineoplastic properties of the essential fatty acids such as linoleic acid have been documented. Linoleic acid and its metabolites have been shown to selectively inhibit and kill a variety of human and animal tumor cells both in vitro and in vivo while affecting normal cells either significantly less or not at all.

Therefore, it was hypothesized that linoleic acid and sesame oil would inhibit the growth of human colon adenocarcinoma and human malignant melanoma cell lines to a greater extent in vitro than their normal counterparts, human colon epithelial cells or human melanocytes. Cells in culture were supplemented with linoleic acid at a dose range of 3-100 $\mu$g/ml. For lipase-digested and undigested sesame oil, a range of 10-300 $\mu$g/ml was used. Growth inhibition was determined by harvesting and counting the total number of cells by hemacytometer after five or eight days of incubation and comparing them to controls. The results showed that free linoleic acid and undigested sesame oil all had a significantly stronger inhibitory effect on both the malignant melanoma and colon adenocarcinoma cell lines than on the corresponding normal cell lines throughout most of the dosage ranges tested.

In comparison with other common vegetable oils and their major component fatty acids, the saturated fatty acid palmitic acid, the monounsaturated fatty acid oleic acid, and the vegetable oils; olive, coconut and safflower all were tested on the malignant melanoma cells. Of these, only safflower, had a significantly selective inhibitory effect.

In conclusion, these results suggest vegetable oils with high linoleic acid content, such as sesame, may possess selective antineoplastic properties against the in vitro growth of malignant melanoma and colon adenocarcinoma. This finding appears to warrant further investigation into the clinical usefulness of the Ayurvedic procedure of topically applying sesame oil.

A comparative risk assessment of chemical genetic engineering, and organic approaches to pest management.

Order No.9933981

As approximately 50% of the world’s food supply is destroyed each year by pests while the human population continues to expand rapidly, agricultural pest management is a global, serious problem. Chemical pesticides, the principal approach to managing pests, has been much analyzed, but relatively little attention has given to organic and genetic engineering methods. The purpose of this study was to conduct a comparative risk assessment of these three approaches, first generally and then in terms of a case study on Roundup Ready soybeans, a genetically modified crop.

The main risk with chemical pesticides is the development of pest resistance. Consequently, a greater percentage of the world’s crops are consumed by pests today than 50 years ago despite a thirty-fold increase in the use of chemicals. When the environmental and health problems of this approach are also considered, it is clear that chemicals not only carry unacceptable risks, they are unsustainable. For this reason the world will likely shift to one of the other two approaches.

The principal general risks of genetic engineering are escape of transgenes into the environment, development of pest resistance, harmful effects on non-target species, continued dependence on chemicals, toxic and allergenic health effects, and increase in antibiotic resistance. The risks of Roundup Ready soybeans in particular are increased use of Roundup, which has been shown to be acutely toxic to a significant number of organisms in the environment and to be potentially carcinogenic to agricultural workers and possibly consumers. Available evidence indicates that all these risks are real and potent. However, because no tests have been conducted to assess any of these risks for the long term, it is recommended that genetically engineered crops should not be commercialized until they are proven safe beyond a reasonable doubt.

The principal risk of the organic approach is the introduction of alien species for biological control, which can also result in effects on non-target species. However, if alien species are avoided, organic agriculture offers the least risk for managing pests effectively while maintaining or even increasing food production without endangering human health or the environment. Source: DAI, 60, no. 06B (1999): p. 2473

Enhancer elements and transcription factors mediating the suppressive effect of IL-1 on CYP1A1 transcriptional activation.

Order No. 9726318

Total P450 content and related activities are known to decrease in cultured rat hepatocytes in response to IL-1 treatment. Previous studies reported that IL-1 suppressed the induction of CYP1A1 and CYP1A2 through a transcriptional mechanism. In order to identify the cis-acting element which mediates IL-1 effect, we analyzed the promoter activity of the 5′-flanking region of CYP1A1 gene. Two elements were identified: xenobiotic responsive element (XRE) and HNF-4 binding site. Transient transfection experiment using primary hepatocytes transfected with CAT reporter genes carrying either XRE1 or XRE2 gave direct evidence that XREs can mediate IL-1 action, although the level of AH receptor binding was not affected. By deletion analysis of the 3.1 kb regulatory region of the CYP1A1 gene, a 36 bp IL-1 responsive region was identified. The region is a composite of three distinct sites: XRE, Sp1-like, and IL-1 responsive element (ILRE). Gel mobility shift assays demonstrated that the ILRE binds constitutively a liver-enriched protein designated as IL-1 responsive protein (ILRP), whose binding activity is reduced by IL-1. Antiserum to the rat HNF-4 transcription factor supershifted the DNA-protein complex formed between ILRE and ILRP. Cotransfection with an HNF-4 expression plasmid increased transcriptional activity of the CYP1A1 minimal promoter carrying one copy of ILRE (about 1.7-fold), or three copies of ILRE (about 2.7-fold) in HepG2 cells. These data suggested that ILRP is in fact HNF-4. Although the transactivation potential of HNF-4 is weak in the context of CYP1A1 promoter, its reduced binding activity upon IL-1 treatment suggests that it may mediate IL-1 action in down- regulating CYP1A1 induction. This is the first report that showed the binding activity of HNF-4 can be down-regulated by IL-1. In summary, IL-1 down-regulation of CYP1A1 transcriptional activation is mediated by XREs, however the mechanism by which this occurs is not known. The HNF-4 binding site may also mediate IL-1 action but more direct evidence is needed. Source: DAI, 58, no. 03B, (1997): 1125

Weight changes of biological and chemical material in a thermodynamically closed system.

Order No. 9318167

The basic research to test the validity of the law of constancy of weight during chemical reactions was done by Landolt /1/, /2/, /3/, Manley /4/. Irregularities in the data of these experiments and the research of Hauschka /6/ on weight changes of sprouting seeds in a thermodynamically closed system suggested a reinvestigation of the question of constancy of weight for chemical reactions and for biological material.The reduction of silver nitrate to metallic silver was chosen as chemical reaction. 3.5 g silver nitrate was reduced in 250-ml gas tight round bottom glass flasks, silver lining the inner glass surface. Sprouting seeds were used in biological experiments. The total weight of the flasks was monitored over several days with an electronic Mettler AE 163 balance, readability 0.1 mg and a mechanical two-pan Volant balance readability 0.2 mg. 25-ml flasks with accordingly smaller silver lining were weighed with and an electronic two-pan Sartorius balance with vacuum capability, readability 1 micro g. Control flasks contained water or glass balls.Extensive artifact research was conducted. Some of the weighings were done under constant temperature and constant relative humidity.Significant deviations from constancy of weight were found in the range of 0.2 to 1.5 mg for the 250-ml flasks and up to 0.06 mg for the 25-ml flasks. For sprouting seeds deviations of the same magnitude were found. Deviations of positive and negative amounts and zero results were obtained. Results were reproduced in blind experiments by five different experimenters. Experiments during eclipses implied a relationship of weight changes with planetary constellations. Source: DAI, 54, no. 03B, (1993): 1449

Transient superinducibility of cyp1a1 mrna and transcription, and the dna elements responsible for mediating this effect

Order No. 9227181

The polycyclic aromatic hydrocarbon (PAH)-inducible CYP1A subfamily of cytochromes P450 is involved in the oxidative metabolism of a wide variety of endogenous and exogenous compounds, including carcinogens and other environmental contaminants. The expression of one member of the subfamily, the CYP1A1 gene, has previously been shown to be under transcriptional control modulated both by PAH-type inducers and by other factors. CYP1A1 transcription can also be superinduced by simultaneous treatment of certain cells in culture with PAH inducers and inhibitors of protein synthesis. In the present study we demonstrated that some cell types that are highly superinducible can be rendered unresponsive to cycloheximide if its addition is delayed approximately 1.5 hours after the cells are exposed to polycyclic aromatic hydrocarbons (Teifeld et al., 1989). This phenomenon, termed transient superinducibility, demonstrates that there are two phases to the initial transcriptional response of the CYP1A1 gene to polycyclic aromatic compounds: an early phase, during which inhibition of protein synthesis can augment the effect of inducers, and a later phase, during which inhibition of protein synthesis does not further increase CYP1A1 gene transcription rate. In order to identify the DNA elements which mediate this phenomenon we used chloramphenicol acetyltransferase (CAT) expression vector analysis.

These studies revealed that the region between $-$1.2 and $- $0.9 kb was necessary to mediate transient superinducibility in the NRK cell line. Further experiments with synthesized CYP1A1 sequences showed that the same elements responsible for mediating the inducible expression of the CYP1A1 gene by xenobiotic treatment, the two xenobiotic response elements, XRE1 and XRE2, were sufficient to mediate both superinduction and transient superinducibility. Loss of responsiveness to cycloheximide was correlated with the disappearance of Ah receptor/XRE binding activity from nuclear extracts of induced cells. Source: DAI, 53, no. 06B, (1992): 2700

Superoxide scavenging, hydrogen peroxide deactivation, and benzoapyrene chemoprotective activities of a Maharishi Ayurveda food supplement, Maharishi Amrit Kalash.

Order No.9427918

Maharishi Ayurveda, a recent restoration of the traditional health care system of India, upholds Maharishi Amrit Kalash (MAK)–an herbal fruit concentrate (MAK-4) and an herbal tablet (MAK-5)–as a rasayana, a food supplement which promotes physiological balance, health, and immunity. Antioxidant and anticarcinogenic activities of MAK have been previously demonstrated in biochemical, cell culture, and in vivo studies. In the present investigation, superoxide scavenging and hydrogen peroxide deactivation properties of MAK were quantified in enzymatic assays, and the ability of MAK to scavenge reactive oxygen species within HeLa cells and to protect C3H/10T1/2 mouse embryo fibroblast-like cells from benzo (a) pyrene transformation was determined.

The superoxide scavenging properties of MAK were investigated with superoxide radicals generated during the catalytic activity of xanthine oxidase. Solutions containing MAK-4 and MAK-S (7.5 and 2.0 mg dry weight of extract/ml) inhibited the reduction of nitroblue tetrazolium (NBT) 96% and 98%, respectively. NBT reduction was decreased 50% by 30 micro g dry weight of extract/ml MAK-4 or 94 micro g dry weight of extract/ml MAK-5. Ascorbic acid inhibition of superoxide radical reduction of NBT reached 88% at 0.176 mg/ml, but declined to 42% at a concentration of 1.76 mg/ml. The rate of uric acid production monitored at 290 nm demonstrated negligible inhibition of xanthine oxidase by MAK-4, MAK-5, or ascorbic acid.

The ability of MAK to deactivate hydrogen peroxide was measured by determining the extent to which MAK inhibited the reduction of scopoletin by H sub 2 O sub 2 generated during the catalytic activity of glucose oxidase. At what appear to be approximately physiological concentrations (0.75 and 0.20 mg dry weight of extract/ml), MAK-4 and MAK-5 inhibited loss of scopoletin fluorescence 79% and 98%, respectively.

In cell culture, extracts from 1 micro g/ml MAK-4 and 10 micro g/ml MAK-5 inhibited the transformation of C3H/10T1/2 cells by benzo(a) pyrene 54% and 56%, respectively. Extracts from 20 micro g/ml MAK-4 and 15 micro g/ml MAK-5 inhibited intracellular reactive oxygen species generated by HeLa cells and monitored by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide reduction 12% and 17%, respectively.

The results contribute to an understanding of previously reported anticarcinogenic, antioxidant, and anti-aging properties of MAK-4 and MAK-5, and warrant consideration in the light of present preventive, nutritional, and chemotherapeutic approaches to health, antioxidant defense, and carcinogenesis. Source: DAI, 55, no. 06B, (1994): 2120

Effects of a multimodal approach of Maharishi consciousness-based health care on carotid

Order No. 3131257

Cardiovascular disease (CVD) is still the largest contributor to morbidity and mortality in the world. Over the past 30 years, focus on primary and secondary prevention of cardiovascular disease and its related risk factors have yielded three major prevention strategies-drug therapies, lifestyle modification and stress reduction therapies. Despite this effort, up to 50% of patients with documented CVD have recurrent cardiac events. A new angle that supports further prevention of CVD is needed. Maharishi Consciousness-Based Health Care system, a natural, prevention-oriented system of health, includes 40 modalities for enlivening the “inner intelligence of the body,” which are responsible for coordinating diverse physiological systems into an integrated whole. This pilot trial compared effects of four Maharishi Consciousness-Based Health Care modalities to those of usual care on carotid intima-media thickness (IMT) in elderly subjects with documented cardiovascular disease and two to six CVD risk factors. The Maharishi Consciousness-Based Health Care modalities included the Transcendental Meditation program, neuro-physiological integration exercises, dietary and herbal supplement approaches. Usual care included the secondary prevention system offered at the University of Iowa Hospitals and Clinics, which are based on practice guidelines promoted by the American Heart Association. Twenty-eight volunteer subjects were matched on age (mean 72 years), gender and severity of documented CVD. Measures were taken for baseline and nine-month posttest. At baseline the experimental group had significantly higher BMI (30 versus 26), triglycerides (177 mg/dl versus 101 mg/dl, and blood pressure (137 versus 120 mm Hg). They were more often single, and had lower income. Covarying for these baseline differences in major CVD risk factors, the experimental group tended to show a greater decrease in mean common carotid IMT after nine months. (Experimental −0.023 mm, Usual Care +0.041 mm, p = 0.07). The IMT regression in the experimental subjects was associated with high compliance. There was a strong correlation between compliance and increases in physical, mental and behavioral strength (r = 0.47), as assessed by Maharishi Consciousness-Based Health Care procedures. These findings suggest that enlivening ‘the body’s inner intelligence’ could be an effective tool to deal with the current epidemic of cardiovascular disease.

The effects of Transcendental Meditation on ambulatory blood pressure, cardiovascular reactivity, anger/hostility, and platelet aggregation

Order No. 9421866

In addition to traditional risk factors, psychosocial stress such as Type A behavior pattern, anger and hostility, and increased cardiovascular reactivity to stress have been proposed as risk factors for cardiovascular disease. Therefore, stress reduction approaches such as Transcendental Meditation (TM) may be useful in modifying these behavioral factors.

Forty normotensive volunteers were pretested and posttested for cardiovascular reactivity to a standard battery of laboratory stressors, underwent ambulatory blood pressure monitoring during the day, and were tested for levels of anger, hostility, and platelet aggregation. They were randomly assigned to either TM or a cognitive-based Stress Education Class (SEC) control group. Both treatment groups involved similar instructional attention and daily practice.

After a four-month treatment period, no significant differences were found between the two treatment groups in cardiovascular reactivity or in average cardiovascular levels in the laboratory or in the field. However, the regular TM practitioners demonstrated increased systolic blood pressure reactivity to the preparation of a speech and to the speech task itself. In addition, the regular TM practitioners also demonstrated a significant reduction in average ambulatory diastolic blood pressure. No significant differences in platelet aggregation, anger or hostility were found between the two groups, except that the SEC group had lower outwardly expressed anger. Among all subjects of the study, significant positive correlations were found between outwardly expressed anger and collagen- induced platelet aggregation, and heart rate reactivity.

These results show that it is possible to decrease average ambulatory blood pressure levels without decreasing cardiovascular reactivity in normotensive subjects with the regular practice of TM. This finding supports the hypothesis that tonic (average) and reactive blood pressure are largely independently regulated and therefore can be differentially modified by behavioral treatment. Since average ambulatory blood pressure is a better predictor of cardiovascular complications of hypertension than clinic blood pressure, this finding may have implications for the prevention of cardiovascular disease. Data from this study also suggest mechanisms whereby stress may be translated into coronary heart disease, i.e., anger may increase coronary heart disease through its association with platelet aggregation and heart rate reactivity. Source: DAI, 55, no. 06B, (1994): 2120

Transcriptional suppression of cytochrome P450 1A1 gene is under redox regulation – Ah receptor-mediated processes with distinct mechanisms.

Order No. 9726319

Oxidative stress in a cell is defined as an unusually high level of reactive oxygen species which can be caused by a number of stimuli. We have investigated the molecular mechanism whereby transcriptional expression of CYP1A1 gene was regulated by redox potential. XRE was found to be the response element by which H(sub 2) O(sub 2) exhibited its inhibitory effect on the transcription of CYP1A1 gene in Hepa 1 cell line using transient transfection technique. However, H(sub 2) O(sub 2) did not alter the DNA binding activity of the Ah receptor. Further study demonstrated that modulation of XRE enhancer strength by various means could modify H(sub 2) O(sub 2)-dependent suppression of CAT expression. The results from this study suggest the presence of a protein that inhibits transactivation by the Ah receptor without influencing its DNA binding ability. In search for the candidate protein(s) which mediated H(sub 2) O(sub 2) action on Ah receptor function, we first demonstrated that overexpression of the product of the retinoblastoma susceptibility gene (RB) downregulated transcription of the CYP1A1 gene. XREs alone were sufficient to mediate RB action. Results from coimmunoprecipitation assays indicated that the Ah receptor coprecipitated with the RB protein or its family member p107 and vice versa. Similar to other RB binding proteins, the Ah receptor only bound to the hypophosphorylated form of RB or p107 protein. To further characterize regulation of CYP1A1 by redox potential, more powerful and more specific oxidants were used to oxidize vicinal sulfhydryl groups in intact Hepa 1 cells. Pretreatment with diamide or phenylarsine oxide for 20 minutes rapidly prevented the formation of ligand-dependent Ah receptor/XRE complex and thus inhibited XRE-mediated luciferase expression. Direct oxidization of the Ah receptor by PAO was further demonstrated by the experiments in which DTT, a reducing agent, could restore the Ah receptor DNA binding activity. Finally, a one hundred-fold difference in the effectiveness between dithiol 2,3-dimercaptopropanol and monothiol 2- mercaptoethanol in reversing PAO-dependent inhibition of Ah receptor DNA binding activity suggests that vicinal sulfhydryl residues may be involved in the DNA binding of Ah receptor. Source: DAI, 58, no. 03B, (1997): 1130