Interindividual differences in the concentration of 1-hydroxypyrene-glucuronide in urine and polycyclic aromatic hydrocarbon-DNA adducts in peripheral white blood cells after charbroiled beef consumption

Biological markers of internal dose and macromolecular dosefrom PAHs provide a potential means of assessing environmentalexposure to PAHs through inhalation, ingestion and percutaneousabsorption. In this study we examined the time course and interindividualvariation of 1-hydroxypyrene-glucuronide (1-OHP-gluc) excretionin urine and PAH—DNA adduct formation in peripheral whiteblood cells (WBCs) after charbroiled (CB) beef consumption.As a marker of internal dose, 1-OHP-gluc was measured in humanurine using immunoaffinity chromatography and synchronous fluorescencespectroscopy. PAH—DNA adducts were measured in WBCs byenzyme-linked immunosorbent assay (ELISA) in order to assessmacromolecular dose. Ten healthy non smoking males consumedidentical amounts of CB beef on five consecutive days. Multipleblood and urine samples were collected before, during, and afterthe feeding period. The morning after the first day of CB beefconsumption, individual urinary concentrations of 1-OHP-glucincreased 10- to 80-fold (range:2.0–16.6 pmol/ml urine)aboveprefeed baseline concentreations (0.23±0.11pmol/ml) inthe 10 subjects. 1-OHP-gluc concentration decreased to nearbaseline levels by 24–72 h after CB beef consumption ended.In contrast, PAH—DNA adducts in WBCs increased markedlyin only four of 10 subjects during or after CB beef consumption.Significant interindividual variation was observed for bothurinary 1-OHP-gluc concentration (P < 0.001 by Kruskal—Wallis)and PAH—DNA adduct levels (P < 0.005) during the feedingperiod. The mean urinary 1-OHP-gluc concentration for each subjectduring and immediately after (days 2–8) the feeding periodwas significantly correlated with their mean PAH—DNA adductlevel in WBCs during the same time period (Spearman r = 0.79,P<0.01). Evidence of segregation of the subjects into separateresponse groups based on level of urinary 1-OHP-gluc was observed,suggesting that discrete determaints may regulate the absorption,metabolism and/or excretion of ingested pyrene.     

Cancer mortality among males in relation to exposures assessed through a job-exposure matrix

To identify potential associations between workplace exposures and cancer mortality risks, job titles collected from 1965 to 1971 for 58,678 men (a subset of a large representative sample of the Canadian workforce) were transformed into probable chemical exposures using a job-exposure matrix developed in Montreal. Mortality follow-up was determined through computerized record linkage with the National Mortality Database in Canada for 1965-1991. Cancer mortality risk was evaluated at two levels of exposure, any and substantial, using Poisson regression controlling for age, calendar period, and social class. Among the 58,678 men, 3,160 died of cancer. Using a liberal reporting criterion, relative risk (RR) >1.0, five or more exposed cancer deaths, p < or = 0.100, several potential associations were identified, including: lung cancer and any exposure to abrasives dust (RR = 2.84), prostate cancer and any exposure to calcium carbonate (RR = 2.46), and prostate cancer and substantial exposure to metallic dust (RR = 2.13).     

Postnatal stress of early weaning exacerbates behavioral outcome in prenatal alcohol-exposed juvenile rats

Some of the behavioral deficits caused by prenatal or postnatal alcohol exposure have been demonstrated to be ameliorated by environmental manipulations such as handling or environmental enrichment. This experiment, in contrast, investigated whether behavioral deficits due to prenatal alcohol exposure could be exacerbated by a stressful experience, early weaning. Pregnant dams were given either a liquid diet with 35% of the calories derived from alcohol, a liquid diet without alcohol to control for any effects of the liquid diet administration, or ad libitum food and water. Half of each litter were weaned at 15 days of age (early weaning) and half were weaned at 21 days of age (normally weaned). Offspring were weighed, tested for activity in an open field at 18 days of age, and trained to find a hidden platform in the Morris water maze at 22-24 days of age. Alcohol-exposed subjects who were weaned early were more impaired in spatial navigation ability than any other group. Similarly, the combination of early weaning and prenatal alcohol exposure caused the slowest growth. All subjects exposed to alcohol, regardless of weaning condition, had greater latencies to find the platform than those from the two control groups. There was no synergistic effect of alcohol and stress on activity levels, but all early-weaned females were more active than normally weaned females; males did not show this effect. Thus, environmental stressors such as early weaning can compound detrimental symptoms of prenatal alcohol exposure. These results have implications for the understanding of the effects of the environment on neuronal plasticity.     

Endothelium-dependent hyperpolarization to acetylcholine in carotid artery of guinea pig: role of lipoxygenase

This study was designed to determine whether lipoxygenase-dependent metabolites of arachidonic acid are involved in the endothelium-dependent hyperpolarization of the guinea pig carotid artery. The membrane potential of vascular smooth muscle cells was measured with intracellular microelectrodes and potassium channels were studied on freshly isolated cells with the patch-clamp technique. Acetylcholine-induced hyperpolarizations were not affected by arachidonyl trifluoromethyl ketone (AACOCF3), quinacrine (phospholipase A inhibitors), or eicosatetraenoic acid (nonspecific inhibitor of lipoxygenase, cytochrome P450, and cyclooxygenase). In contrast, cinnamyl-3,4 dihydroxy-alpha-cyanocinnamate (CDC) and AA861 (lipoxygenase inhibitors) as well as 1-(6-(17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino) hexyl)-1H-pyrrole-2,5-dione (U-73122) (phospholipase C inhibitor) produced a significant inhibition of the hyperpolarization. An opener of intermediate conductance calcium-activated potassium channels, 1-ethyl-2-benzamidazolinone (1-EBIO), induced a hyperpolarization that was unaffected by AACOCF3, CDC, AA861, or U-73122 but was inhibited by charybdotoxin. (+/-)12-hydroxy-eicosatetraenoic acid (12-HETE) and 12(S)-hydroperoxy-eicosatetraenoic acid (12(S)-HpETE) did not induce any significant changes in membrane potential. CDC inhibited the voltage-gated potassium current and increased the large conductance calcium-activated potassium current whereas AA861 inhibited both potassium currents. These results confirm that, in the isolated carotid artery of the guinea pig, stimulation of endothelial muscarinic receptors involves phospholipase C activation and indicate that the activation of phospholipase A2 and the release of lipoxygenase metabolites is unlikely to explain endothelium-dependent hyperpolarization.     

EDHF: bringing the concepts together

Endothelial cells synthesize and release vasoactive mediators in response to various neurohumoural substances (e.g. bradykinin or acetylcholine) and physical stimuli (e.g. cyclic stretch or fluid shear stress). The best-characterized endothelium-derived relaxing factors are nitric oxide and prostacyclin. However, an additional relaxant pathway associated with smooth muscle hyperpolarization also exists. This hyperpolarization was originally attributed to the release of an endothelium-derived hyperpolarizing factor (EDHF) that diffuses to and activates smooth muscle K(+) channels. More recent evidence suggests that endothelial cell receptor activation by these neurohumoural substances opens endothelial cell K(+) channels. Several mechanisms have been proposed to link this pivotal step to the subsequent smooth muscle hyperpolarization. The main concepts are considered in detail in this review.     

A group of glutamatergic interneurons expressing high levels of both neurokinin-1 receptors and somatostatin identifies the region of the pre-Bötzinger complex

The pre-B?tzinger complex (pre-B?tC) is a physiologically defined group of ventrolateral medullary neurons that plays a central role in respiratory rhythm generation. These cells are located in a portion of the rostral ventrolateral medulla (RVLM) that is difficult to identify precisely for lack of a specific marker. We sought to determine whether somatostatin (SST) might be a marker for this region. The rat pre-B?tC area was defined as a 500-microm-long segment of ventrolateral medulla coextensive with the ventral respiratory group. This region was identified by juxtacellular labeling of neurons with respiratory-related activity and by its location rostral to the phrenic premotor neurons. It contained most of the SST-ir neuronal somata of the RVLM. These cells were small (107 microm(2)) and expressed high levels of preprosomatostatin mRNA. They were strongly neurokinin 1 receptor (NK1R)-ir and were selectively destroyed by saporin conjugated with an NK1R agonist (SSP-SAP). Most SST-ir neurons (>90%) contained vesicular glutamate transporter 2 (VGLUT2) mRNA, and terminals immunoreactive for SST and VGLUT2 protein were found in their midst. Few SST-ir neurons contained GAD-67 mRNA (<1%) or preproenkephalin mRNA (6%). Retrograde labeling experiments demonstrated that over 75% of the SST-ir neurons project to the contralateral pre-B?tC area, but none projects to the spinal cord. In conclusion, the RVLM contains many neurons that express preprosomatostatin mRNA. A subgroup of these cells contains high levels of SST and NK1R immunoreactivity in their somata. These glutamatergic interneurons identify a narrow region of the RVLM that appears to be coextensive with the pre-B?tC of adult rats.     

Hypothalamic orexin (hypocretin) neurons express vesicular glutamate transporters VGLUT1 or VGLUT2

Initially recognized for their importance in control of appetite, orexins (also called hypocretins) are neuropeptides that are also involved in regulating sleep, arousal, and cardiovascular function. Loss of orexin appears to be the primary cause of narcolepsy. Cells expressing the orexins are restricted to a discrete region of the hypothalamus, but their terminal projections are widely distributed throughout the brain. With the diversity of function and broad distribution of orexin terminals, it is not known whether the orexin cells constitute a homogeneous population. Because orexins produce neuroexcitatory effects, we hypothesized that orexin-containing neurons are glutamatergic. In the present study we used digoxigenin-labeled cRNA probes for the vesicular glutamate transporters, VGLUT1 and VGLUT2, for in situ hybridization studies in combination with immunohistochemical detection of orexin cell bodies in the hypothalamus. In general, cells in the hypothalamus expressed low levels of the vesicular glutamate transporters relative to other areas of the forebrain, such as the cortex and thalamus. Light labeling for VGLUT2 mRNA was detected in about 50% of the orexin-immunoreactive neurons, and a much smaller percentage (approximately 13%) of orexin-immunoreactive cells was found to express VGLUT1. Despite the fact that intense labeling for GAD67 mRNA was found in a large number of cells throughout the hypothalamus, none of the orexin-immunoreactive cells was found to be GABAergic. These findings, showing that many of the orexin neurons are glutamatergic, are consistent with the neuroexcitatory effects of orexin but suggest that another neurochemical phenotype may define the remaining subset of orexin neurons.     

Self-healing juvenile cutaneous mucinosis

A healthy 14-month-old black girl presented with a 3-week complaint of "knots" on the face and hands. The lesions were acute in onset and asymptomatic. Multiple, firm, nontender, skin-colored to erythematous nodules were noted on the scalp, forehead, axillae, lower legs, abdomen, and hands. A skin biopsy specimen revealed a well-circumscribed accumulation of mucin in the reticular dermis. Colloidal iron stain was positive. Radiographs showed soft tissue prominence only. Serum protein electrophoresis, thyroid function tests, complete blood count, sedimentation rate, and antinuclear antibody were normal, except for lymphocytosis. Findings were consistent with self-healing juvenile cutaneous mucinosis (SHJCM). SHJCM is a condition of unknown etiology characterized by rapid onset of asymptomatic, indurated papules or nodules. Affected children may have arthralgias, but are otherwise well. Spontaneous resolution is the rule. Most skin lesions in our patient had resolved within 6 months of onset. This patient is unique because of the young age of onset.     

Comparison of the cost and effectiveness of two strategies for maintaining hepatitis B immunity in hemodialysis patients

A decision-analysis model was developed to compare the strategies to maintain hepatitis B virus (HBV) immunity in hemodialysis patients who responded to the primary HBV vaccine. Our hypothesis is that the routine, annual administration of the vaccine booster to all hemodialysis patients (non-screening strategy) is more cost-effective than the current strategy of vaccination based on anti-HBs titers (screening strategy). Under baseline assumptions, the screening strategy was less costly and was associated with fewer HBV infections than the non-screening strategy. The results of our model did not support our hypothesis, and indicate that regularly screening patients for HBV immunity before revaccination is less costly and more effective than the empiric vaccination of hemodialysis patients.