Alteration of estrogen-regulated gene expression in human cells induced by the agricultural and horticultural herbicide glyphosate.

Gene expression is altered in mammalian cells (MCF-7 cells), by exposure to a variety of chemicals that mimic steroid hormones or interact with endocrine receptors or their co-factors. Among those populations chronically exposed to these endocrine disruptive chemicals are persons, and their families, who are employed in agriculture or horticulture, or who use agricultural/horticultural chemicals. Among the chemicals most commonly used, both commercially and in the home, is the herbicide glyphosate. Although glyphosate is commonly considered to be relatively non-toxic, we utilized in vitro DNA microarray analysis of this chemical to evaluate its capacity to alter the expression of a variety of genes in human cells. We selected a group of genes, determined by DNA microarray analysis to be dysregulated, and used quantitative real-time PCR to corroborate their altered states of expression. We discussed the reported function of those genes, with emphasis on altered physiological states that are capable of initiating adverse health effects that might be anticipated if gene expression were significantly altered in either adults or embryos exposed in utero.

Postimpoundment time course of increased mercury concentrations in fish in hydroelectric reservoirs of northern Manitoba, Canada.

Mercury (Hg) concentrations in fish in boreal reservoirs have been shown to be increased for up to 3 decades after impoundment. However, the time course of increased concentrations is not well known. The purpose of this study was to determine the evolution of Hg concentrations in fish in the boreal reservoirs of northern Manitoba, Canada, and its relationship with severity of flooding. We determined total Hg concentrations in three species of fish for up to 35 years after impoundment in 14 lakes and lake basins. Postimpoundment trends depended on fish species and reservoir. In the benthivorous lake whitefish (Coregonus clupeaformis), Hg concentrations increased after flooding to between 0.2 and 0.4 microg g(-1) wet weight compared with preimpoundment concentrations between 0.06 and 0.14 microg g(-1) and concentrations in natural lakes between 0.03 and 0.06 microg g(-1). Hg concentrations in lake whitefish were usually highest within 6 years after lake impoundment and took 10 to 20 years after impoundment to decrease to background concentrations in most reservoirs. Hg concentrations in predatory northern pike (Esox lucius) and walleye (Sander vitreus) were highest 2 to 8 years after flooding at 0.7 to 2.6 microg g(-1) compared with preimpoundment concentrations of 0.19 to 0.47 microg g(-1) and concentrations in natural lakes of 0.35 to 0.47 microg g(-1). Hg concentrations in these predatory species decreased consistently in subsequent years and required 10 to 23 years to return to background levels. Thus, results demonstrate the effect of trophic level on Hg concentrations (biomagnification). Peak Hg concentrations depended on the amount of flooding (relative increase in lake surface area). Asymptotic concentrations of approximately 0.25 microg g(-1) for lake whitefish and 1.6 microg g(-1) for both walleye and northern pike were reached at approximately 100% flooding. Downstream effects were apparent because many reservoirs downstream of other impoundments had higher Hg concentrations in fish than would be expected on the basis of flooding amount.

Alteration of gene expression in human cells treated with the agricultural chemical diazinon: possible interaction in fetal development.

Agricultural chemicals frequently alter human health or development, typically because they have endocrine agonist or antagonist activities and alter hormone-regulation of gene expression. The insecticide, diazinon, was evaluated for gene expression disrupting activity using MCF-7 cells, an estrogen-dependent human cell line, to examine the capacity of the insecticide to disrupt gene expression essential for morphological development, immune system development or function, and/or central nervous system development and function. MCF-7 cells were treated with 30, 50 or 67 ppm diazinon, and gene expression was measured in treated cells compared to expression in untreated or estrogen-treated cells. DNA microarray analysis of diazinon-treated cells showed significant up- or down-regulation of a large number of genes compared to untreated cells. Of the 600 human genes on the Phase 1 chip utilized for these studies, two specific genes--calreticulin and TGF-beta3--were selected for corroboration using quantitative real time PCR (qrtPCR). qrtPCR, completed to assess gene expression levels for calreticulin and TGFbeta3, confirmed results showing significant up-regulation of these two genes obtained from the microarray data. These studies were designed to provide baseline data on the gene expression-altering capacity of a specific chemical, diazinon, and allow a partial assessment of the potentially deleterious effects associated with exposure of human cells to this chemical. Currently, it is not known whether results from cells in vitro can be extrapolated to human health consequences of chemical exposure.

Altered gene expression in human cells treated with the insecticide diazinon: correlation with decreased DNA excision repair capacity.

Many industrial and agricultural chemicals have steroid hormone agonist or antagonist activities and disrupt hormone-regulated gene expression. The widely-used agricultural insecticide, diazinon, was evaluated using MCF-7 cells - a breast cancer-derived, estrogen-dependent, human cell line - to examine the capacity of this chemical to alter steroid hormone-regulated gene expression. MCF-7 cells were treated with 30, 50, or 67 ppm of diazinon, and gene expression in treated cells was measured as mRNA levels in the cells compared to mRNA levels in untreated or estrogen-treated cells. DNA microarray analysis showed significant up- or down-regulation of a number of genes in treated cells compared to untreated cells. Of the 600 human genes on the chip utilized, specific genes with related functions were selected for additional consideration. Real time quantitative PCR (qrtPCR) completed to corroborate mRNA levels as a measure of specific gene expression, confirmed results obtained from analysis of the micro-array data. The data show that ERCC5, encoding Xeroderma pigmentosum protein G (XPG), essential for DNA excision repair, and ribonucleotide reductase subunit M1 (RNRM1), encoding a gene necessary for providing the nucleotides needed for DNA repair, were down-regulated in cells treated with diazinon. These studies were designed to provide base-line data on the gene expression-altering capacity of a specific agricultural chemical, diazinon, and allow assessment of some of the potentially deleterious effects associated with exposure of human cells to diazinon.

Effects of environmental and maternally derived methylmercury on the embryonic and larval stages of walleye (Stizostedion vitreum).

The effects of environmental and maternally derived methylmercury (MeHg) on the embryonic and larval stages of walleye (Stizostedion vitreum) were investigated using eggs collected during two successive spawning seasons. Eggs were collected from fish in a mercury (Hg)-polluted environment (Clay Lake, Ontario, Canada), and from fish in two relatively pristine lakes (Lakes Manitoba and Winnipeg, in the province of Manitoba). Both bioaccumulation of Hg into muscle and its mobilization into eggs was significantly higher in Clay Lake females. Maternal muscle MeHg concentration was positively correlated with female length and egg MeHg was positively correlated with muscle MeHg concentration in all three populations. Hatching success of eggs from all three stocks declined significantly with increasing waterborne MeHg (0.1-7.8 ng l-1). Hatching success was not significantly affected by egg MeHg concentration. Embryonic heart rate declined with increasing waterborne MeHg concentration, but larval growth was not affected. Occurrence of larval deformities was negatively correlated with size of female, but was not significantly correlated with MeHg in either eggs or water. Larval MeHg was positively correlated with the concentrations of MeHg in eggs demonstrating transmission of MeHg from females. Uptake of ambient MeHg was higher in larvae exposed to higher waterborne MeHg concentrations.

Intra- and interpopulation variability in maternal transfer of mercury to eggs of walleye (Stizostedion vitreum).

Mercury concentrations were determined for unfertilized eggs from seven walleye populations and for muscle and liver tissue from three of these seven populations in Canada and the United States. Female walleye transferred very little of their body methylmercury burden to their eggs. Methylmercury concentrations in eggs were 1.1-12% of those in muscle, and methylmercury burdens in eggs represented only 0.2-2.1% of the total body burden. Egg methylmercury as a percentage of total mercury increased with maternal length across populations. Percent methylmercury also increased with egg total mercury concentration but the slope of this relationship varied among populations. Egg methylmercury concentration increased with female age, and both muscle and liver methylmercury concentrations. Egg methylmercury concentration was also affected by female length at age but the effect of this relationship varied among populations. Mean predicted egg methylmercury concentrations (ng g(-1) dry) of 8-year-old, 550-mm females for the seven populations were as follows: Clay Lake, 796; Lake Superior, 24.3; Lake Winnipeg, 16.3; Lake Erie, 11.8; Primrose Lake, 8.76; Lake Manitoba, 7.32; Waconda Lake, 6.69.

Uptake of mercury by fish in an experimental boreal reservoir.

We studied the uptake of mercury (Hg) by finescale dace (Phoxinus neogaeus) in an experimental boreal reservoir for 2 years prior to flooding and 3 years after flooding, and in a natural wetland pond over the same 5-year period. Hg uptake was much higher after flooding as compared to uptake before flooding and in the natural pond. After flooding, Hg concentrations in late summer were usually 2-3x higher than concentrations observed prior to flooding. Net uptake of Hg by fish over the summer in the experimental reservoir was 0.25 and -0.07 microg per fish in the 2 years before flooding as compared to 0.63, 0.64, and 0.42 microg per fish in the 3 years after flooding. Thus, Hg uptake by fish responded quickly to flooding and was highest in the first 2 years following impoundment. Uptake in the reference pond ranged from 0.10 to 0.28 microg of Hg per fish over the same 5-year period. Calculated fluxes of Hg on an areal basis ranged from 0.04-0.09 microg m-2 year-1 in the reference pond, were 0.08 and -0.02 microg m-2 year-1 in the experimental reservoir prior to flooding, and ranged from 0.14-0.22 microg m-2 year-1 in the experimental reservoir after flooding. These fluxes were much smaller than fluxes of methyl mercury (MeHg) through the zooplankton and emerging insect communities. Most (71-89%) of the mercury measured in the muscle of finescale dace was MeHg, and the proportion that was MeHg decreased over the summer period prior to flooding, but increased over the summer after flooding. Growth of fish was not significantly affected by flooding. Fish ate predominantly benthic invertebrates (64-84% of food items found in stomachs), with lesser proportions of crustacean zooplankton (16-31% of items found in stomachs) and feeding was similar after as compared to before flooding. Therefore, differences in Hg uptake did not appear to be the result of changes in diet.

Computerized self-assessment of psychosis severity questionnaire [COSAPSQ] in schizophrenia: preliminary results.

We attempted to develop and validate a computer-driven patient self-rated questionnaire [COSAPSQ] which should provide a reliable, rapid, and inexpensive method to assess symptom severity in patients with psychosis in general and with schizophrenia in particular. After giving informed consent patients with DSM-IV schizophrenia or schizoaffective disorder were interviewed and rated on PANSS and CGI. Subsequently patients completed the COSAPSQ questionnaire (61 multiple choice questions) in the presence of an observer. The analysis of the first 29 rating sets showed that patients with CGI scores of 3-6 completed the questionnaire in a mean time of 21.6 minutes. One-way analysis of variance of COSAPSQ total scores by CGI ratings was highly significant (p < .001). COSAPSQ total scores correlated well with PANSS total, general and positive scores and with CGI (all r = 6-.7; p < .005). The next versions of the questionnaire will require some adjustments: overall fewer questions, improved assessment of negative symptoms, and improved graphic presentation.

Constricted expressiveness and psychophysiological reactivity in schizophrenia.

The present study investigated flat affect in schizophrenia within a multicomponent model of emotions in which affects were identified with the expression or display of feelings. Display, as well as other components of emotion, such as subjective experience and psychophysiological reactivity, were examined as 24 schizophrenic inpatients (12 with flat affect) and 12 depressed inpatients were guided through imagery of personal emotional experiences. It was hypothesized that display variables would show a relationship with flat affect, whereas physiological and subjective variables would not. Increased flatness of affect was associated with longer pauses and reduced dyadic interaction and less zygomatic (cheek) electromyogram activity. The flat affect patients unexpectedly showed more corrugator (brow) electromyogram activity compared with the depressed group, which perhaps reflects difficulty in self-expression. The flat affect group had faster heart rates at baseline. The groups did not, however, differ in self-report of the intensity of experienced emotion. These results support the view that flat affect patients show a reduction in emotional display, whereas other subsystems of emotion processing appear normally engaged.

Pursuit eye movements in chronic schizophrenics: relationship between increased saccades and negative symptoms.

Smooth pursuit eye movements of chronic schizophrenics (n = 16) and controls (n = 10) were recorded while subjects tracked a sinusoidally-moving target. Negative symptoms in schizophrenics were rated using the Schedule for the Assessment of Negative Symptoms. An increased frequency of smooth pursuit eye movement abnormalities was found in the schizophrenic group, including greater variability of gain and more frequent and larger saccades. Frequency of saccades was positively correlated with the Negative Symptoms scales for attentional impairment and alogia. The results confirm findings of eye movement dysfunction in schizophrenics and provide new evidence that the motor and cognitive dysfunctions of schizophrenia may be related.

The effects of electromyographic feedback training on suppression of the oral-lingual movements associated with tardive dyskinesia.

The efficacy of electromyographic feedback training in reducing the magnitude and frequency of the oral-lingual movements associated with tardive dyskinesia (TD) was investigated in a groups design. Twenty adult male inpatients diagnosed as having TD using the Abnormal Involuntary Movements Scale (AIMS) were randomly assigned to one of two treatment conditions. Following identification, all participants were initially reduced to the lowest effective dosage of neuroleptics, and then discontinued from anticholinergics. Following one month on this regimen, they were given a course of feedback training consisting of ten 14-minute sessions. Group one participants were provided with a tone contingent upon oral-lingual movements above a yoked threshold. Group two participants were given noncontingent feedback tones generated randomly. Weekly AIMS were administered as well as an initial baseline during each session to determine current level of oral-lingual activity. An analysis of session effects indicated significantly more suppression of oral-lingual activity in the contingent group versus the noncontingent feedback group. Jaw and forehead activity also measured showed reductions of similar magnitudes for both groups.

Competency-based advancement program for pharmacists.

A competency-based advancement program for practicing pharmacists is presented. The criteria for selecting this method of providing incentives for pharmacists are outlined. Procedures for developing and implementing this competency-based program are described. Five competency levels for pharmacists were developed based on the pharmacist's educational background, training, and experience. Differentiated position titles and competency expectations for each level are summarized. Criteria for pharmacist placement and promotion from one completely level to another are outlined. The administration of the program is described, including budget considerations and methods for evaluating its effectiveness. This system recognizes and rewards accomplishments of staff pharmacists without removing them from patient care roles.

Third-party reimbursement for pharmacist instruction about antihemophilic factor.

The trial and evaluation of pharmacy education program to teach a 14-year-old hemophiliac to self-administer Factor VIII at home is described. The home therapy program realized a cost saving of $20,230 for this patient for the first year; school attendance increased 15%. The results of the trial were used to justify a professional fee from Blue Cross of Central Ohio for the pharmacist's educational services.