The immediate effects of cervical spine manipulation on pain and biochemical markers in females with acute non-specific mechanical neck pain: a randomized clinical trial.

Study Design: Randomized clinical trial with pre-test, post-test control group design. Objectives: To examine the immediate effects of cervical spinal manipulation (CSM) on serum concentration of biochemical markers (oxytocin, neurotensin, orexin A, and cortisol). Background: Several studies have found an association between spinal manipulation (SM) and pain perception. However, the mechanism by which SM modulates pain remains undefined. Methods: Twenty-eight female subjects with non-specific mechanical neck pain were randomly assigned to one of two interventions (CSM versus sham CSM). Blood samples were drawn before and immediately after the respective interventions. Oxytocin, neurotensin, orexin A, and cortisol were measured from the blood and serum using the Milliplex Map Magnetic Bead Panel Immunoassay on the Luminex 200 Platform. Results: In the CSM group, there were significant increases in pre- versus post-manipulation mean oxytocin (154.5 ± 60.1 vs. 185.1 ± 75.6, p = .012); neurotensin (116.0 ± 26.5 vs.136.4 ± 34.1, p < . 001); orexin A (52.2 ± 31.1 vs. 73.8 ± 38.8, p < .01) serum concentration; but no significant differences in mean cortisol (p = .052) serum concentration. In the sham group, there were no significant differences in any of the biomarkers (p > .05). Conclusion: The results of the current study suggest that the mechanical stimuli provided through a CSM may modify neuropeptide expression by immediately increasing the serum concentration of nociception-related biomarkers (oxytocin, neurotensin, orexin A, but not cortisol) in the blood of female subjects with non-specific mechanical neck pain.

Substantial changes in gene expression level due to the storage temperature and storage duration of human whole blood.

Blood is a valuable clinical sample for high-throughput analysis of gene expression and is likely to become more popular as a diagnostic tool and as a predictive measure of disease progression and drug responsiveness. Gene expression data from blood that has been stored at ambient temperature for greater than 1 h vs. blood samples that have been lysed immediately post-collection shows dramatic changes in relative gene expression for a number of cytokines, chemokines, and transcription factors. Results indicate significant changes in the relative expression of several genes, many of which were either up-regulated or down-regulated, because of storage at ambient temperature: (1) In only 4 h of storage at ambient temperature, greater than 10-fold increases in relative gene expression were observed for interleukin-8 (IL-8), c-myc, and c-fos; (2) Up-regulation of IL-8, a chemokine that mediates inflammatory cell migration, took place only 1-h after collection and increased nearly 100-fold by 4 h; (3) Down-regulation of several anti-inflammatory genes was observed for blood stored at ambient temperature; and (4) A general trend toward selective enhancement of inflammatory responses was observed, mediated by possible mRNA transcription and turnover. These results validate the need for the rapid lysis of whole blood after removal from the source.

Modulation of neuroimmune parameters during the eustress of humor-associated mirthful laughter.

CONTEXT: Humor therapy and the related mirthful laughter are suggested to have preventive and healing effects. Although these effects may be mediated by neuroendocrine/neuroimmune modulation, specific neuroimmune parameters have not been fully investigated. OBJECTIVE: To determine the efficacy of mirthful laughter to modulate neuroimmune parameters in normal subjects. DESIGN: A series of 5 separate studies based on a multivariate repeated measures design, with post hoc simple contrast analysis. SETTING: The schools of medicine and public health at Loma Linda University, Loma Linda, Calif. SUBJECTS: 52 healthy men. INTERVENTION: Viewing of a humor video for 1 hour. Blood samples were taken 10 minutes before, 30 minutes into, and 30 minutes and 12 hours after the intervention. MAIN OUTCOME MEASURES: Natural killer cell activity; plasma immunoglobulins; functional phenotypic markers for leukocytes including activated T cells, nonactivated T cells, B cells, natural killer cells, T cells with helper and suppressor markers, and assessment of plasma volume and compartmental shifts; plasma cytokine--interferon-gamma; and total leukocytes with subpopulations of lymphocytes, granulocytes, and monocytes. RESULTS: Increases were found in natural killer cell activity (P < .01); immunoglobulins G (P < .02), A (P < .01), and M (P < .09), with several immunoglobulin effects lasting 12 hours into recovery from initiation of the humor intervention; functional phenotypic markers for leukocyte subsets such as activated T cells (P < .01), active cytotoxic T cells (P < .01), natural killer cells (P = .09), B cells (P < .01), helper T cells (P < .02), uncommitted T cells with helper and suppressor markers (P < .02), helper/suppressor ratio (P = .10) with several leukocyte subset increase effects lasting 12 hours after the humor experience; the cytokine interferon-gamma (P = .02), with increases lasting 12 hours; total leukocytes (P < .05), with specific subpopulation lymphocytes during the intervention (P < .01) and 90 minutes into recovery (P < .05); and granulocytes during the intervention (P < .05) and 90 minutes following the intervention (P < .01). CONCLUSION: Modulation of neuroimmune parameters during and following the humor-associated eustress of laughter may provide beneficial health effects for wellness and a complementary adjunct to whole-person integrative medicine therapies.

Composite effects of group drumming music therapy on modulation of neuroendocrine-immune parameters in normal subjects.

CONTEXT: Drum circles have been part of healing rituals in many cultures throughout the world since antiquity. Although drum circles are gaining increased interest as a complementary therapeutic strategy in the traditional medical arena, limited scientific data documenting biological benefits associated with percussion activities exist. OBJECTIVE: To determine the role of group-drumming music therapy as a composite activity with potential for alteration of stress-related hormones and enhancement of specific immunologic measures associated with natural killer cell activity and cell-mediated immunity. DESIGN: A single trial experimental intervention with control groups. SETTING: The Mind-Body Wellness Center, an outpatient medical facility in Meadville, Pa. PARTICIPANTS: A total of 111 age- and sex-matched volunteer subjects (55 men and 56 women, with a mean age of 30.4 years) were recruited. INTERVENTION: Six preliminary supervised groups were studied using various control and experimental paradigms designed to separate drumming components for the ultimate determination of a single experimental model, including 2 control groups (resting and listening) as well as 4 group-drumming experimental models (basic, impact, shamanic, and composite). The composite drumming group using a music therapy protocol was selected based on preliminary statistical analysis, which demonstrated immune modulation in a direction opposite to that expected with the classical stress response. The final experimental design included the original composite drumming group plus 50 additional age- and sex-matched volunteer subjects who were randomly assigned to participate in group drumming or control sessions. MAIN OUTCOME MEASURES: Pre- and postintervention measurements of plasma cortisol, plasma dehydroepiandrosterone, plasma dehydroepiandrosterone-to-cortisol ratio, natural killer cell activity, lymphokine-activated killer cell activity, plasma interleukin-2, plasma interferon-gamma, the Beck Anxiety Inventory, and the Beck Depression Inventory II. RESULTS: Group drumming resulted in increased dehydroepiandrosterone-to-cortisol ratios, increased natural killer cell activity, and increased lymphokine-activated killer cell activity without alteration in plasma interleukin 2 or interferon-gamma, or in the Beck Anxiety Inventory and the Beck Depression Inventory II. CONCLUSIONS: Drumming is a complex composite intervention with the potential to modulate specific neuroendocrine and neuroimmune parameters in a direction opposite to that expected with the classic stress response.

Dietary intake and biochemical, hematologic, and immune status of vegans compared with nonvegetarians.

Dietary and nutritional status of individuals habitually consuming a vegan diet was evaluated by biochemical, hematologic, and immunologic measures in comparison with a nonvegetarian group. On the basis of 4-d dietary records, the intake of female and male vegans tended to be lower in fat, saturated fat, monounsaturated fat, and cholesterol and higher in dietary fiber than that of vegetarians. With computed food and supplement intakes, vegan diets provided significantly higher amounts of ascorbate, folate, magnesium, copper, and manganese in both female and male participants. The body mass index (BMI; in kg/m(2)) of the vegans was significantly lower than that of the nonvegetarians and 9 of the 25 vegans had a BMI <19. Serum ferritin concentrations were significantly lower in vegan men but iron and zinc status did not differ between the sexes. Mean serum vitamin B-12 and methylmalonic acid concentrations did not differ; however, 10 of the 25 vegans showed a vitamin B-12 deficit manifested by macrocytosis, circulating vitamin B-12 concentrations <150 pmol/L, or serum methylmalonic acid >376 nmol/L. Vegans had significantly lower leukocyte, lymphocyte, and platelet counts and lower concentrations of complement factor 3 and blood urea nitrogen but higher serum albumin concentrations. Vegans did not differ from nonvegetarians in functional immunocompetence assessed as mitogen stimulation or natural killer cell cytotoxic activity.

The impact of caffeine use on tobacco cessation and withdrawal.

Continuous caffeine consumption with smoking cessation has been associated with more than doubled caffeine plasma levels. Such concentrations may be sufficient to produce caffeine toxicity symptoms in smoking abstinence conditions. To test whether caffeine abstinence influences smoking cessation, 162 caffeine-using smokers were enlisted from American Lung Association smoking cessation programs. Volunteers were randomly assigned by clinic to caffeine-use and caffeine-abstinence conditions and measured for 3 weeks post-smoking cessation, at 6 months and one year. Results showed a significant linear increase in caffeine sputum levels across 3 weeks post cessation for those who quit smoking and continued using caffeine. Three weeks after cessation, concentrations reached 203% of baseline for the caffeine user. Typical nicotine withdrawal symptoms occurred during the first 16 days of cessation. The caffeine abstainers, but not continued users of caffeine, reported increased fatigue during the first 3 days of cessation. Among complete caffeine abstainers, compared with caffeine users, there was a significant increase in fatigue, a decrease in stimulation, and a marginal increase in caffeine craving immediately following tobacco cessation. There were no differences between the groups on other withdrawal symptoms or in cessation success at 16 days, 6 months, or 12 months.

Indapamide regresses, but transdermal clonidine does not regress, left ventricular hypertrophy in hypertensive diabetic patients.

This report describes the effects of indapamide versus transdermal clonidine on left ventricular hypertrophy (LVH) in hypertensive diabetic patients. A sample of 24 hypertensive diabetic men, aged 40-68 years, with echocardiographically proven LVH was equally divided in to 2 groups. Group 1 was treated with indapamide 2.5 mg/day, and group C with transdermal clonidine weekly. Left ventricular mass and posterior wall and septal thickness were measured by standard echocardiograms done at baseline and every 6 months. At 24 months, treatment crossover was done. Normotension was maintained throughout the study. With indapamide, LVH regression was measurable at 6 months, and left ventricular mass had returned to normal after 18 months. Transdermal clonidine did not regress LVH, but when the patients were switched to indapamide, LVH did regress. Clonidine maintained normal ventricular dimensions after regression had been induced by indapamide.

Effects of exogenous monoiodotyrosine on the serum levels of anterior pituitary hormones.

Monoiodotyrosine is a tyrosine hydroxylase inhibitor. Ingestion of one gram monoiodotyrosine caused a 10,000-fold increase of serum monoiodotyrosine from basal levels of 0.69 +/- 0.20 nmol/l to a peak of 10.6 +/- 1.7 mumol/l in women and 7.1 +/- 2.3 mumol/l in men 30 min later, and the t1/2 was 45 min. Monoiodotyrosine stimulated PRL to a peak of 170 +/- 51 micrograms/l in women and 90 +/- 6 micrograms/l in men 30 min after the monoiodotyrosine peak, or 60 min after the ingestion. Other anterior pituitary hormones were unchanged. Dopamine infusion or L-dopa pretreatment attenuated the monoiodotyrosine effect. TRH exaggerated the PRL peak, and chlorpromazine did not increase but prolonged the hyperprolactinemia. These results suggest that dopamine synthesis inhibition may be the mechanism of PRL stimulation.

Modulation of the renin-aldosterone system by iodotyrosines as tyrosine hydroxylase inhibitors.

This study shows that MIT and DIT stimulate aldosterone secretion. This may be due to their tyrosine hydroxylase inhibitory property. Dopamine abolishes the stimulation. Prolonged MIT administration enhances the stimulation of aldosterone secretion and can cause hypokalemia. Volume expansion reverses the hyperaldosteronism. PRA and blood pressure do not change, even after prolonged MIT intake.

The effect of long endurance running on natural killer cells in marathoners.

Ten experienced marathoners were exercised 3 h in the laboratory. Blood samples were collected at 0 h baseline, 1 h exercise, and 5 min, 1.5 h, 6 h, and 21 h recovery and were analyzed for total number of lymphocytes expressing membrane antigens found on natural killer (NK) cells. NK activity was also measured. Four of the seven subpopulations of lymphocytes studied, Leu-11+19+, Leu-11+19-, Leu-11+7-, and Leu-19+11-, showed significant within-subject effects over time, using repeated measures ANOVA. Simple contrasts with baseline values showed that, at 1.5 h and 21 h recovery, total number of lymphocytes bearing three different combinations of NK markers, Leu-11+19+, Leu-11+19-, and Leu-11+7-, were significantly decreased when compared with baseline values. At 1.5 h recovery, NK activity was significantly decreased below baseline levels for four of the six effector NK cell/target K562 myelogenous leukemia cell (E:T) ratios tested. At 6 h recovery, NK activity was still decreased significantly with the 12.5:1 and 3:1 E:T ratios. By 21 h recovery, NK activity did not differ significantly from baseline levels. Cortisol levels at 5 min post-exercise were negatively correlated with NK activity at 1.5 h recovery (r = -0.62, P = 0.05, 50:1 E:T ratio; r = -0.66, P = 0.04, 25:1 E:T ratio). Further research is needed to elucidate the effect these changes have on host immunosurveillance and immunoresponsiveness in vivo.

Extrathyroidal physiology of monoiodotyrosine in humans.

Normal serum monoiodotyrosine (MIT) levels (n = 152) were 0.69 +/- 0.20 nmol/l. There was wide variation of MIT levels in a 24-hour period without diurnal pattern, and there was no change throughout the menstrual cycle. MIT levels declined upon aging, but levels in hypo- and hyperthyroidism were not significantly different. MIT levels were detected in athyrotic patients (0.32 +/- 0.08 nmol/l). Desiccated thyroid raised the athyrotic MIT levels to the normal range, while levothyroxine did not. Diiodotyrosine (DIT) infusion caused an MIT rise which paralleled but lagged 1 h behind the DIT rise. These data suggest thyroidal as well as nonthyroidal sources of MIT, one of which is deiodination of DIT. Ingestion of 1 g MIT increased serum MIT to 10.6 +/- 1.7 mumol/l in women, and 7.1 +/- 2.3 mumol/l in men 30 min after ingestion; the serum half-life was 45 min.

Neuroendocrine and stress hormone changes during mirthful laughter.

Positive emotional activities have been suggested as modifiers of neuroendocrine hormones involved in the classical stress response. To detect changes in these components during a mirthful laughter experience, the authors studied 10 healthy male subjects. Five experimental subjects viewed a 60 minute humor video and five control subjects did not. Serial blood samples were measured for corticotropin (ACTH), cortisol, beta-endorphin, 3,4-dihydrophenylacetic acid (dopac)--the major serum neuronal catabolite of dopamine, epinephrine, norepinephrine, growth hormone, and prolactin. Repeated measures analysis of variance showed that cortisol and dopac in the experimental group decreased more rapidly from baseline than the control group (p = 0.011, p = 0.025, respectively). Epinephrine levels in the experimental group were significantly lower than the control at all time points (p = 0.017). Growth hormone levels in the experimental group significantly increased during baseline (p = 0.027) and then decreased with laughter intervention (p less than 0.0005), whereas, the controls did not change over time (p = 0.787). ACTH, beta-endorphin, prolactin, and norepinephrine levels did not significantly increase. The mirthful laughter experience appears to reduce serum levels of cortisol, dopac, epinephrine, and growth hormone. These biochemical changes have implications for the reversal of the neuroendocrine and classical stress hormone response.

Effects of long-endurance running on immune system parameters and lymphocyte function in experienced marathoners.

The extent and duration of changes in leukocyte subsets, lymphocyte subpopulations, spontaneous blastogenesis, cortisol, and catecholamines were measured in ten experienced marathoners, who ran 3 h to exhaustion in a laboratory setting. Blood samples were taken at baseline, 1 h of exercise, and 5 min, 1.5 h, 6 h, and 21 h of recovery. The 3-h endurance run was associated with significant leukocytosis, granulocytosis, neutrophilia, monocytosis, and eosinopenia during recovery. All of these parameters except for eosinophils returned to normal by 21 h of recovery. Total lymphocyte count increased 31% at 1 h of exercise, then decreased 19% at 1.5 h of recovery when compared with baseline values. T cell count showed no significant changes, but B cell lymphocytosis was measured at 5 min and 6 h of recovery. T helper/T suppressor ratio (H/S) was significantly elevated 39% at both 1.5 h and 21 h of recovery due to the decrease in number of T suppressor cells. Spontaneous blastogenesis was significantly increased 52% by 1 h of exercise and remained elevated throughout recovery. The increase in cortisol from baseline to 1.5 h of recovery correlated positively with the increase in both total leukocyte count (r = 0.78, P = 0.008) and granulocyte count (r = 0.81, P = 0.005). Our results suggest that exhaustive endurance exercise in marathon runners is associated with many significant perturbations in immune system parameters, most of which return to normal levels at 21 h of recovery.

Complement and immunoglobulin levels in athletes and sedentary controls.

Eleven marathon runners (42.7 +/- 2.1 yrs, 54.2 +/- 1.8 ml.kg-1.min-1) and nine sedentary controls (44.2 +/- 1.2 yrs, 33.3 +/- 1.1 ml.kg-1.min-1) were studied during 30 min of rest, a graded maximal treadmill test using the Balke protocol, and 45 min of recovery to determine the effects of training and acute exercise on complement and immunoglobulin levels. Three baseline and five recovery blood samples were obtained in addition to repeated 5-min samples during exercise. Data for the exercise period were analyzed using a multiple regression approach to repeated measures ANOVA to allow comparison between groups on a percent VO2max basis. Groups did not differ during any of the three phases for IgG, IgA, or IgM. Resting levels of complement C3 (0.89 +/- 0.05 vs 1.27 +/- 0.10 g/L, P less than 0.001) and C4 (0.19 +/- 0.02 vs 0.29 +/- 0.03 g/L, P less than 0.001) were significantly lower in athletes than in controls. Exercise complement C3 [F(1,18) = 14.1, P = 0.001] and C4 [F(1,18) = 7.6, P = 0.013], and recovery complement [F(1,18) = 19.4, P less than 0.001] and C4 [F(1,18) = 13.5, P = 0.002] were also lower in the athletes than in sedentary controls. Acute increases during exercise were not associated with changes in catecholamines or cortisol. These data suggest that blood concentrations of C3 and C4, but not IgG, IgA, or IgM, are decreased during rest, graded maximal exercise, and recovery in marathon runners in comparison with sedentary controls.

Evaluation of the powder-formulated enzyme multiplied immunoassay technique quantitative single test for gentamicin.

The usefulness of the enzyme multiplied immunoassay quantitative single test (EMIT QST) gentamicin assay was assessed for gentamicin analysis in patient sera. The EMIT QST reagents are in powder form in a single, premeasured vial and are run on a thermoregulated sample processor that controls mixing and timing steps. The results of the clinical evaluation showed that the standard curve was stable throughout a 26-day study period. Within-run precision on 20 replicates at 4.0 micrograms/ml yielded a coefficient of variation (CV) of 5.6%; between-run precision on 66 analyses at 6.0 micrograms/ml over a 152-day period yielded a CV of 4.0%. Mean recovery through the range of the standard curve with 10 spiked patient samples was 102%. Comparative analysis with radioimmunoassay of 95 patient samples showed a correlation of 0.97, with y = 0.93x - 0.03. It was concluded that the EMIT QST gentamicin assay is an appropriate, rapid methodology for patient gentamicin analysis.

Simple, rapid 125I-labeled cyclosporine double antibody/polyethylene glycol radioimmunoassay used in a pediatric cardiac transplant program.

We modified the Sandoz cyclosporine radioimmunoassay because of our need for frequent clinical monitoring of cyclosporine drug levels in allo- and xenograft pediatric cardiac transplant patients. With application of a commercially available [125I]cyclosporine label in place of [3H]cyclosporine and a second antibody/polyethylene glycol (PEG) method of separation in place of charcoal separation, we simplified and enhanced the speed and precision of assay performance. Studies of 140 whole blood samples comparing this new method to the [3H]cyclosporine radioimmunoassay (RIA) method of Berk and colleagues yielded a coefficient of correlation of 0.96 (p less than 0.00001) with means of 626 and 667 ng/ml for [3H]RIA and [125I]RIA, respectively, and a regression equation of y = 28 + 1.02x. The major advantages are that total assay time is reduced to approximately 1 h; [125I]cyclosporine label is used, avoiding the problems associated with liquid scintillation counting; and precision is enhanced by separating bound and free fractions with second antibody/PEG. These modifications should provide for greater ease of assay performance and improved clinical utility of cyclosporine monitoring not only in the pediatric but also in the adult transplant patient.

Immunoassays: a roadmap for the 1980s.

The use of immunoassay has greatly expanded the capability of the clinical laboratory. This paper describes the basic principles behind immunoassay as well as a number of specific applications of those principles. Radioimmunoassay, enzyme immunoassay, fluorescent immunoassay, chemiluminescence, and nephelometry are some of the subjects discussed. The discussion of fundamental principles forms a basis for exploration of the future of immunoassay.