Isolation and characterization of human osteoblasts from needle biopsies without in vitro culture.

SUMMARY: We isolate and characterize osteoblasts from humans without in vitro culture. These techniques should be broadly applicable to studying the pathogenesis of osteoporosis and other bone disorders. INTRODUCTION: There is currently no data regarding the expression of specific genes or pathways in human osteoblasts that have not been subjected to extensive in vitro culture. Thus, we developed methods to rapidly isolate progressively enriched osteoblast populations from humans and characterized these cells. METHODS: Needle bone biopsies of the posterior iliac crest were subjected to sequential collagenase digests. The cells from the second digest were stained with an alkaline phosphatase (AP) antibody, and the AP+ cells were isolated using magnetic cell sorting. RESULTS: Relative to AP- cells, the AP+ cells contained virtually all of the mineralizing cells and were enriched for key osteoblast marker genes. The AP+ cells were further purified by depletion of cells expressing CD45, CD34, or CD31 (AP+/CD45/34/31- cells), which represented a highly enriched human osteoblast population devoid of hematopoietic/endothelial cells. These cells expressed osteoblast marker genes but very low to undetectable levels of SOST. We next used high-throughput RNA sequencing to compare the transcriptome of the AP+/CD45/34/31- cells to human fibroblasts and identified genes and pathways expressed only in human osteoblasts in vivo, but not in fibroblasts, including 448 genes unique to human osteoblasts. CONCLUSIONS: We provide a detailed characterization of highly enriched human osteoblast populations without in vitro culture. These techniques should be broadly applicable to studying the pathogenesis of osteoporosis and other bone disorders.

Effects of estrogen therapy on bone marrow adipocytes in postmenopausal osteoporotic women.

UNLABELLED: One-year treatment of osteoporotic postmenopausal women with transdermal estrogen resulted in significant decreases in bone marrow adipocyte volume and prevented increases in adipocyte number as compared to placebo-treated controls. Estrogen treatment also prevented increases in mean adipocyte size over 1 year. INTRODUCTION: Aging is associated not only with bone loss but also with increases in bone marrow adipocytes. Since osteoblasts and adipocytes are derived from a common precursor, it is possible that with aging, there is a preferential "switch" in commitment of this precursor to the adipocyte over the osteoblast lineage. We tested the hypothesis that the apparent "age-related" increase in marrow adipocytes is due, at least in part, to estrogen (E) deficiency. METHODS: Reanalysis of bone biopsies from a randomized, placebo-controlled trial involving 56 postmenopausal osteoporotic women (mean age, 64 years) treated either with placebo (PL, n = 27) or transdermal estradiol (0.1 mg/d, n = 29) for 1 year. RESULTS: Adipocyte volume/tissue volume (AV/TV) and adipocyte number (Ad#) increased (by 20%, P < 0.05) in the PL group, but were unchanged (Ad#) or decreased (AV/TV, by -24%, P < 0.001) in the E group. E treatment also prevented increases in mean adipocyte size over 1 year. CONCLUSIONS: These findings represent the first in vivo demonstration in humans that not only ongoing bone loss, but also the increase in bone marrow adipocyte number and size in postmenopausal osteoporotic women may be due, at least in part, to E deficiency.

Utility of type I procollagen propeptide assays for assessing abnormalities in metabolic bone diseases.

We measured type I procollagen carboxyl-terminal propeptide (PICP) by a commercial radioimmunoassay and amino-terminal propeptide (PINP) by an enzyme-linked immunosorbent assay (ELISA) developed in our laboratory in serum from 75 normal women, 10 growing girls, 84 normal men, and 197 patients with various metabolic bone diseases. The molar concentrations of serum PINP were 100-fold higher than those of PICP, suggesting differences in the metabolism of PICP and PINP. In normal women, serum PICP values correlated positively with age and serum PINP values correlated negatively with age (r = 0.28 and -0.32, respectively; P = 0.02). In normal men, however, PICP correlated negatively with age (r = -0.32, P = 0.003) whereas PINP did not change. As assessed by Z scores (SD from age- and sex-specific predicted normal mean), changes in serum PICP and PINP values were concordant in hypoparathyroidism (mean Z scores for PICP and PINP, -0.63 and -1.48, respectively) and Cushing's syndrome (0.50 and 0.40) but were discordant in acromegaly (0.78 and -0.68), hyperthyroidism (1.33 and -0.66), untreated postmenopausal osteoporosis (-0.11 and 0.40), fluoride-treated postmenopausal osteoporosis (-0.61 and 1.08), Paget's disease (4.05 and -0.20), and chronic renal failure (1.45 and -0.50). With either assay, deviations from normal were less pronounced than the deviations of concurrently measured serum osteocalcin and bone alkaline phosphatase values. The deviations in these latter two values agreed better with those of PICP than with those of PINP, except in untreated or fluoride-treated osteoporotic patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Circulating levels of cytokines that modulate bone resorption: effects of age and menopause in women.

Interleukin 1 alpha (IL-1 alpha), interleukin 1 beta (IL-1 beta), and interleukin 6 (IL-6) are cytokines with potent bone-resorbing effects; some of these biologic effects are opposed by interleukin-1 receptor antagonist (IL-1ra). In vitro and animal model studies suggest that these cytokines are paracrine mediators of the increased bone resorption associated with estrogen deficiency, and increases in their production also could contribute to age-related bone loss. Therefore, we measured serum concentrations of these cytokines in 80 normal women who were 24-87 years old. IL-6 concentration correlated highly with age (p < 0.001) and increased three-fold during life. However, multiple-regression analysis showed no significant correlation between serum IL-6 levels and menopausal status, serum estradiol concentration, or markers for bone turnover (serum bone alkaline phosphatase, osteocalcin, carboxyl-terminal telopeptide of type I collagen, or 24 h urinary free pyridinoline excretion). Serum IL-1 alpha, IL-1 beta, or IL-1ra level did not change with age and, by multiple-regression analysis, did not correlate with markers of bone turnover, except IL-1ra weakly with ICTP. We found no relationship between bone-resorbing cytokines and ovarian function. Although the large age-related increase in serum IL-6 concentration could contribute to age-related bone loss, the lack of correlation with markers for bone turnover argues against this. However, based on the strong evidence in experimental animals that these cytokines are involved in estrogen action on bone, further studies in humans are warranted.

Circulating cytokine levels in osteoporotic and normal women.

Interleukin-1 (IL-1) and IL-6 have been postulated to play roles in the pathogenesis of postmenopausal osteoporosis. To test this hypothesis, we measured circulating levels of IL-6, IL-1 alpha, and IL-1 beta in 40 age-matched normal and 40 osteoporotic women with vertebral fractures and increased bone turnover. Since IL-1 activity is modulated by the IL-1 receptor antagonist (IL-1ra), we also measured circulating IL-1ra levels in these women. Despite having higher rates of bone turnover as assessed by bone biochemical markers, the osteoporotic women had serum levels of IL-6, IL-1 alpha, and IL-1 beta that were similar to the normal women. IL-1ra levels (mean +/- SEM) tended to be lower in the osteoporotic women (143 +/- 21 pg/mL) vs. the normal women (189 +/- 22 pg/mL; P = 0.08). The IL-1 alpha/IL-1ra ratio, an index of IL-1 alpha bioactivity, was higher in the osteoporotic women (6.4 +/- 1.1) than in the normal women (4.4 +/- 0.5; P < 0.05). There were nonsignificant trends for increased IL-1 beta/IL-1ra and (IL-1 alpha + IL-1 beta)/IL-1ra ratios in the osteoporotic vs. normal women. None of the cytokines correlated with markers of bone resorption in either group of women. In summary, although there was a trend for a higher IL-1/IL-1ra ratio in the osteoporotic women, we were unable to demonstrate unequivocal abnormalities of cytokines affecting bone resorption in peripheral serum of women with postmenopausal osteoporosis. However, it is possible that increased production of these cytokines occurs in the local environment of bone or bone marrow and is not detected by analysis of peripheral serum. Thus, further studies will be required to exclude this possibility.

Concurrent assays of circulating bone Gla-protein and bone alkaline phosphatase: effects of sex, age, and metabolic bone disease.

We measured the serum concentrations of 2 biochemical markers of bone formation, bone Gla-protein (BGP) and bone alkaline phosphatase (BAP), in 164 normal subjects and 164 patients with metabolic bone disorders. The data were reported as Z scores (deviation in SDs from the sex-specific age regression in normal subjects). Both serum BGP and BAP distinguished abnormalities well (mean Z scores for BGP and BAP, respectively) and gave concordant results in patients with hypoparathyroidism (-1.7, -1.4), hyperthyroidism (+1.1, +1.8), primary hyperparathyroidism (+3.6, +2.5), acromegaly (+1.2, +2.8), and postmenopausal osteoporosis (+0.4, +1.9). The 2 markers gave discordant results, however, in patients with glucocorticoid excess (-2.4, +0.9), Paget's disease (+1.8, +41.8), chronic renal failure (+16.3, +0.4), and osteolytic metastases (-1.4, +5.9). These discrepancies may have occurred because serum BGP and BAP concentrations reflect different aspects of osteoblast function or because there are differences in their clearance from the circulation. Consequently, more information is derived about the level of bone formation across the wide range of metabolic bone disorders when both biochemical markers are assayed.

Sensitization to early life stress and response to chemical odors in older adults.

This study examined the hypothesis that older persons who currently report illness from environmental chemical odors (cacosmia) may have experienced higher levels of stress early in life than did noncacosmic controls. The hypothesis derives from a time-dependent sensitization (TDS) model for cacosmia (Bell et al 1992) that predicts a relative interchangeability of stress and chemicals in inducing and eliciting sensitized responses in vulnerable individuals. Subjects were selected from those in the top 24% (cacosmic) and bottom 27% (noncacosmic) of a sample of 192 older adults (mean age 73.8 years) for self-reported frequency of illness form the odors of pesticide, car exhaust, paint, perfume, and new carpet. As in previous investigations, cacosmics were younger, more depressed, and more shy; cacosmics also included a higher proportion of women (83% versus 61%). As predicted, cacosmics rated themselves higher in stress for the first four decades of their lives, but not the recent past or present, even after controlling for depression, anxiety, hostility, shyness, age, and gender. Cacosmics reported increased prevalence of physician-diagnosed nasal allergies, breast cysts, hypothyroidism, sinusitis, food sensitivities, irritable bowel, and migraine headache. Only 4% of the overall sample (including 9% of the cacosmics) acknowledged the controversial physician diagnosis of "chemical sensitivity." The replicated observation of greater shyness in cacosmics is consistent with the ability of hyperreactivity to novelty to predict enhanced susceptibility to TDS from low levels of pharmacological agents in animals. The findings support a TDS model for cacosmia and suggest that cacosmia as a symptom identifies a large subset of the nonindustrial population with significant psychophysiological health problems that merit further objective examination.

A polysomnographic study of sleep disturbance in community elderly with self-reported environmental chemical odor intolerance.

Subjective sleep complaints and food intolerances, especially to milk products, are frequent symptoms of individuals who also report intolerance for low-level odors of various environmental chemicals. The purpose of the present study was to evaluate the objective nature of nocturnal sleep patterns during different diets, using polysomnography in community older adults with self-reported illness from chemical odors. Those high in chemical odor intolerance (n = 15) exhibited significantly lower sleep efficiency (p = .005) and lower rapid-eye-movement (REM) sleep percent (p = .04), with a trend toward longer latency to REM sleep (p = .07), than did those low in chemical intolerance (n = 15), especially on dairy-containing as compared with nondairy (soy) diets. The arousal pattern of the chemical odor intolerant group differed from the polysomnographic features of major depression, classical organophosphate toxicity, and subjective insomnia without objective findings. The findings suggest that community elderly with moderate chemical odor intolerance and minimal sleep complaints exhibit objectively poorer sleep than do their normal peers. Individual differences in underlying brain function may help generate these observations. The data support the need for similar studies in clinical populations with chemical odor intolerance, such as multiple chemical sensitivity patients and perhaps certain veterans with "Persian Gulf Syndrome."

Demography of handedness in two samples of randomly selected adults (N = 2083).

Handedness was examined in relation to sex, race, age, education, occupation, marital status, and religious preferences for two random samples of adults drawn from an urban population. There were statistically significant differences on each variable for the total of 2083 respondents as well as for many subgroups based on combinations of sex, race, and age. The results are compared to previous findings.

Bone resorption in healthy and osteoporotic postmenopausal women: comparison markers for serum carboxy-terminal telopeptide of type I collagen and urinary pyridinium cross-links.

We compared two highly specific markers for bone resorption-pyridinium cross-links (pyridinoline (PYR) and deoxypyridinoline (DPR)) in urine and carboxy-terminal telopeptide of type I collagen (ICTP) in serum-in 63 healthy postmenopausal women and 63 women with osteoporosis characterized by more bone resorption than bone formation. The ICTP, PYR and DPR levels were all higher, by 24% (p = 0.001), 16% (p = 0.05) and 25% (p = 0.004), respectively, in the osteoporotic women. For the merged groups, there were significant correlations between serum ICTP concentration and urinary PYR (r = 0.667, p < 0.0001) and DPR (r = 0.452, p < 0.0001) excretion; for the osteoporotic and normal women separately, the r values were 0.73 (p < 0.01) and 0.45 (p < 0.01) for PYR and 0.51 (p < 0.01) and 0.22 (p = 0.08) for DPR versus ICTP respectively. Weak correlations in linear regression between ICTP and various indices of bone formation (osteocalcin, bone-specific alkaline phosphatase and carboxy-terminal propeptide of type I procollagen) disappeared when the correlation between ICTP and pyridinolines was accounted for by calculation of partial correlation coefficients in multiple regression analysis. Serum ICTP concentration appears to discriminate between groups of normal and osteoporotic women as well as urinary pyridinium cross-links, which is thus far the most sensitive method for assessing bone resorption.

Medical histories and psychological profiles of middle-aged women with and without self-reported illness from environmental chemicals.

BACKGROUND: Cacosmia, which is a predictor of cognitive deficits in industrial samples, is a core symptom of several controversial syndromes. Previous studies of cacosmic populations have considered only psychiatric but not medical or family histories of identified patients. METHOD: This questionnaire survey study examined subjective characteristics of illness from chemical odors, sensitivity to chemicals, psychological and stress profiles, and medical, psychiatric, and family health histories of 28 middle-aged women with cacosmia in self-reported poor health attributed to chemicals (MCS), 17 controls with cacosmia in good health, and 20 normal controls without cacosmia in good health. RESULTS: Those with MCS rated themselves in significantly poorer overall health with higher Pennebaker symptom scores, a larger number of chemical triggers, and greater frequency of illness from chemicals than the other two groups, even after controlling for variables on which the groups differed (i.e., education, Symptom Checklist-90 [revised] somatization, obsessive-compulsiveness, depression, anxiety, phobic anxiety, psychoticism, Barsky Somatic Symptom Amplification, and Cheek-Buss shyness). Despite increased levels of affective distress, those with MCS reported the greatest intolerance for alcohol and the lowest alcohol consumption. CONCLUSION: The data suggest that women with MCS report increased disability, multiple medical diagnoses including inflammatory and gynecologic dysfunctions, and psychological distress. The data are consistent descriptively with the phenomenology of somatization disorder. However, the persisting significance of group health rating differences after controlling for psychological variables, the lack of differences in life stress ratings between those with MCS and healthy cacosmics, the later age at onset (60% after age 30 years), and the lack of excess family psychiatric histories in this sample of women with MCS suggest a potential role for an organic factor in the evolution of poor health in certain cacosmics.

Significance of T-lymphocytes in wound healing.

To determine the importance of T-lymphocytes in wound healing, we examined the effect of T-lymphocyte depletion on the healing of surgical wounds. Thirty Balb/c mice were injected intraperitoneally with 1 mg of rat anti-mouse (IgG2b) cytotoxic monoclonal antibody (30H12) against the Thy1.2 (all T) determinant. Twenty-four hours later animals showed a greater than 95% depletion of Thy1.2 cells in peripheral blood and spleen. Thirty control mice received nonspecific rat immunoglobulin (1 mg). Twenty-four hours after treatment mice underwent a 2.5 cm dorsal skin incision with subcutaneous placement of polyvinyl alcohol sponges. Injections were repeated at weekly intervals. Wound healing was assessed at 2, 3, and 4 weeks by the breaking strength of wound strips and by the hydroxyproline content of sponge granulomas (an index of wound reparative collagen deposition). Thy1.2 depletion at death was 95% to 57% in peripheral blood and 86% to 68% in the spleen. Both groups gained weight equally. We found that T cell depletion significantly impairs wound breaking strength and wound collagen deposition at all times studied. The data strongly suggest that T-lymphocytes modulate fibroblast activity during normal wound healing.

Effect of oxotremorine on local cerebral glucose utilization in motor system regions of the rat brain.

The [14C]2-deoxy-D-glucose technique was used to test the effects of central muscarinic stimulation on local cerebral glucose utilization (LCGU) in rats. Systemic administration of the muscarinic agonist oxotremorine (OXO, 0.7 mg/kg, ip) increased LCGU in brain regions involved in motor function: the sensorimotor cortex; the extrapyramidal motor system, including the striatum, globus pallidus, red nucleus, substantia nigra, subthalamus and ventral nucleus of the thalamus; the cerebellar vermis, fastigial nucleus and nucleus interpositus; and the vestibular nucleus. No effects were observed in the fibers of the pyramidal tract, internal capsule, cerebellar white matter and the dentate nucleus. Increases were not affected by methylatropine (1 mg/kg sc), but were completely antagonized by scopolamine (2.5 mg/kg, ip). The anatomic distribution and magnitude of the LCGU response to OXO were not simply correlated with the reported densities of muscarinic receptors.

Sex differences in the effects of dexamethasone phosphate on behavior in rats.

The effect of dexamethasone phosphate (DEX) administered in rats' drinking water on running activity and open field behavior was investigated. In Experiment 1 males were given DEX continuously from either five days or one day prior to and throughout testing. Only 5 day treatment significantly increased running wheel activity. DEX had no significant effect on males' 4 day open field activity, but significantly reduced open field and home cage defecation. In Experiment 2 females given DEX defecated significantly more in the open field than controls. This effect on females does not appear to be due to a general metabolic change, since DEX females, like males, defecated significantly less than controls in the home cage. Females' open field activity was not significantly affected. Weight loss and plasma corticosterone analysis confirmed the effectiveness of the dosage used. There appears to be a sex difference in the effects of DEX on open field defecation, possibly due to interaction with gonadal hormones.

Sex difference in the effect of dexamethasone on open-field behavior in rats: gonadal hormones.

PIP: A study was undertaken to determine whether or not the sex differences in the effect of dexamethasone (DEX) on open-field defecation depended on the difference in circulating gonadal hormones in normal male and female rats. The experiment involved ovariectomized females, ovariectomized females given supplementary male hormones, and sham-operated females. In comparison to control groups, all the DEX-treated groups showed a suppression of plasma corticosterone levels, a decrease in weight levels, and reduced adrenal weights. Open-field defecation increased in the DEX-treated female rats but open-field activity was not affected. No differences in behavior or physiological changes were evident between the 3 differently treated groups. These results indicate that sex differences in the response of open-field defecation to DEX-treatment is not the result of sex differences in circulating sex hormones. The response may spring from the organizing effects of the gonadal hormones during the perinatal stage rather than the activating effect of gonadal hormones during adulthood.^ieng

Trait shyness in the elderly: evidence for an association with Parkinson's disease in family members and biochemical correlates.

The emergence of potential treatments to slow the progression of idiopathic Parkinson's disease (PD) has increased the need for early identification of persons at risk. Although considered controversial, some prior studies indicate that PD patients may have premorbid histories of greater trait introversion or shyness as well as increased rates of disorders associated with shyness (e.g., anxiety, affective disorders, and irritable bowel syndrome). Essential features of trait shyness include (a) inhibited and avoidant behaviors and (b) physiological hyperreactivity to the novel or unfamiliar. In parallel, (a) depression in PD patients is associated with increased harm avoidance (a possible serotonergic function), and (b) PD patients have premorbid and comorbid decreases in novelty-seeking (a possible dopaminergic function). Taken together, previous research suggests the following hypotheses: (1) given evidence for marked heritability of shyness, shy elderly should report higher rates of PD in their family members than would nonshy elderly; and (2) shy elderly without PD should exhibit psychological and biologic characteristics similar to those reported in PD. Two groups, representing the top 27% (n = 37) and bottom 31% (n = 43) of scores on a standardized shyness scale, were drawn from a larger cohort of 138 older adults (ages 50-90) living in an active retirement community. Seventeen percent of the shy versus 2% of the nonshy reported PD in a family member or self (P < .05). Shy elderly were significantly more anxious (P < .01) and depressed (P < .05) than were the nonshy.(ABSTRACT TRUNCATED AT 250 WORDS)

Assessing beliefs of older rural Americans about nutrition education: use of the focus group approach.

We used the focus group interview technique as a preliminary research step in developing a nutrition education intervention for rural seniors who, because of less than optimal eating habits and changing demographics, are an important target audience. Sixty-eight well, active, rural North Dakota seniors, 60 years or older, from communities of 2,500 or fewer people, participated in five focus groups conducted in late summer 1988. As a qualitative research approach, focus group interviews offer a means of obtaining in-depth information on a specific topic from representatives of a target audience in a discussion group atmosphere. Focus groups require careful preparation and structuring and should include a capable moderator, a prepared discussion guide, carefully recruited participants, and a comfortable setting. The process generated ideas that we are using to develop a health promotion nutrition intervention that will be a mailed-home approach, including use of incentives, social role models, cholesterol screening, and learning activities. The intervention relies on the interest and ability of seniors to make positive health changes. We conclude that the focus group approach is useful in developing nutrition education interventions.

Chronic methadone administration to male rats: tolerance to adverse effects on sires and their progeny.

Previous studies have shown that acute administration of methadone to male rats prior to mating results in adverse effects on their progeny, particularly decreased birth weights and increased neonatal mortality. Rather than chronic administration accentuating these effects, results of the present study indicate that tolerance developed so that no adverse effects were found in offspring sired after 21--32 days of methadone administration. In the sire, maintenance of normal weights of accessory sex glands after 4 months of daily methadone suggests that tolerance developed to the CNS effect(s) responsible for the depressed serum LH and testosterone levels found after acute administration of narcotics. In contrast, tolerance did not develop to the inhibition of weight gain produced by methadone administration. No evidence for a dominant lethal effect could be found after chronic methadone administration, in contrast to suggestive evidence for this effect found in previous experiments after acute methadone administration.

Quantitative EEG patterns during nose versus mouth inhalation of filtered room air in young adults with and without self-reported chemical odor intolerances.

Individuals who report illness (e.g. nausea, headache) from common chemical odors tend to report CNS symptoms suggestive of olfactory-limbic system involvement. This study compared the resting quantitative electroencephalographic (qEEG) patterns of young adult college students reporting subjectively elevated chemical odor intolerance ratings (HICI) with those of controls reporting little or no odor intolerance (LOCI). Each group was subdivided into those with higher (HIDEP) vs. lower (LODEP) ratings of concomitant depression. Nineteen channels of EEG were recorded during a single session over four separate rest periods, respectively, following baseline, cognitive, chemical exposure and olfactory identification tests. Each recording involved two 30-s, eyes-closed, filtered room air breathing conditions: (1) nose inhalation followed by mouth exhalation and (2) mouth inhalation followed by mouth exhalation. HICI showed significantly less beta 1 (beta 1) over the temporal-central region during nose than during mouth inhalation. Over some temporal and central leads, task, DEP and CI interacted to influence beta 1 as well. For theta (theta), CI differences emerged during nose inhalation after the cognitive task at Cz, after chemical exposures at C3, Cz and C4 and after the olfactory ID task at C4. CI differences emerged during mouth breathing after the olfactory ID task at Cz, C4 and T4. The T5-T6 coronal array showed significant CI differences after chemical exposures during nose breathing and during mouth breathing after the cognitive and olfactory ID tasks. The theta findings in the HICI may be related to reports of disturbed attention in CI.

Self-reported illness from chemical odors in young adults without clinical syndromes or occupational exposures.

The present survey of young adult college students investigated the prevalence of self-reported illness from the smell of the five following common environmental chemicals (cacosmia): (1) pesticide, (2) automobile exhaust, (3) paint, (4) new carpet, and (5) perfume. Sixty-six percent of 643 students reported feeling ill from one or more of the five chemicals; 15% identified the smell of at least four chemicals as making them ill. Ratings of illness from pesticide correlated weakly but significantly with ratings for the largest number of individual symptoms (9 of 11); daytime tiredness and daytime grogginess both correlated at high levels of significance with illness ratings (on a 5-point scale) for four of the five chemicals. The most cacosmic group (CS) included significantly more women (79%) than the noncacosmic group (NS) (49%); women overall were more cacosmic than men (p < .001), even with the significant covariate of depression. Ratings of cacosmia correlated only weakly with scores for depression (r = 0.16), anxiety (r = 0.08), and trait shyness (r = 0.18) in the total sample. On stepwise multiple regression with cacosmia score as the dependent measure, shyness accounted for 5.8% of the variance, while depression, anxiety, sense of mastery, and repression did not enter the equation. Histories of physician-diagnosed hay fever, but not asthma, were more frequent in the CS (16%) than in the NS group (5%). Without the confounds of chronic illness or specific treatment programs, these data are similar to patterns described clinically for a subset of patients with multiple chemical sensitivities (MCS), including previous data on increased nasal resistance in MCS.(ABSTRACT TRUNCATED AT 250 WORDS)