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Self-reported exposures and their association with unexplained illness in a population-based case-control study of Gulf War veterans.

Many factors have been considered as possible causes of the unexplained illness reported by veterans of the Gulf War (GW). In this study, we report an analysis of risk factors and unexplained illness in a population-based sample of GW veterans who underwent clinical evaluation. Multiple risk factors were compared in 241 veterans who met criteria for unexplained illness and 113 healthy controls. Results suggest that GW unexplained illness is most highly associated with combat conditions, heat stress, and having sought medical attention during the GW. When controlling for multiple simultaneous exposures during the GW, interactions around pyridostigmine bromide, insecticides and repellents, and stress were not significant. These results indicate that most unexplained illness in GW veterans cannot be explained by neurotoxic effects of exposures to chemicals that inhibit cholinesterase activity.

Adult↗

Behavioral sensitization after repeated formaldehyde exposure in rats.

Multiple chemical sensitivity (MCS) is a phenomenon whereby individuals report increased sensitivity to chemicals in the environment, and attribute their sensitivities to prior exposure to the same or often structurally unrelated chemicals. A leading hypothesis suggests that MCS is akin to behavioral sensitization observed in rodents after repeated exposure to drugs of abuse or environmental stressors. Sensitization occurring within limbic circuitry of the central nervous system (CNS) may explain the multisymptom complaints in individuals with MCS. The present studies represent the continuing development of an animal model for MCS, the basis of which is the CNS sensitization hypothesis. Three behaviors were assessed in rats repeatedly exposed to formaldehyde (Form) inhalation. In the first series of experiments, rats were given high-dose Form exposure (11 parts per million [ppm]; 1 h/day x 7 days) or low-dose Form exposure (1 ppm; either 1 h/day x 7 days or 1 h/day x 5 days/week x 4 weeks). Within a few days after discontinuing daily Form, cocaine-induced locomotor activity was elevated after high-dose Form or 20 days of low-dose Form inhalation. Approximately 1 month later, cocaine-induced locomotor activity remained significantly elevated in the 20-day Form-exposed rats. The second experiment assessed whether prior exposure to Form (20 days, as above) would alter the ability to condition to an odor (orange oil) paired with footshock. The results suggested a tendency to increase the conditioned fear response to the odor but not the context of the footshock box, and a decreased tendency to extinguish the conditioned fear response to odor. The third experiment examined whether CNS sensitization to daily cocaine or stress would alter subsequent avoidance responding to odor (Form). Daily cocaine significantly elevated approach responses to Form, while daily stress pretreatment produced a trend in the opposite direction, producing greater avoidance of Form. Preliminary studies indicated that repeated daily Form inhalation (20 days, as above) produced a greater avoidance to subsequent Form presentation, suggesting that daily Form inhalation may serve as a stressor. The results support the hypothesis that repeated chemical exposure in rats may produce CNS plasticity manifest as greater sensitivity to dopaminergic drugs, enhanced fear conditioning to odor paired with an aversive event, and greater avoidance of odors. Some of these behavioral changes observed in rats may provide a link with symptoms in a subset of individuals with MCS.

Administration, Inhalation↗

Pathophysiologic effects of acceleration stress in the miniature swine.

Four groups of six animals each were exposed to simulated +Gz aerial combat maneuvers (SACM). The first group was exposed to 5 SACM/d for 1 d. The other three groups were exposed to 5 SACM/d three times per week for 1 week, 1 month, and 6 months, respectively. The SACM was 100 s long and contained two 5-s +9Gz peaks. A control group of four animals was exposed to 1 d of five 100-s +1Gz exposures. All animals were unanesthetized. The animals were necropsied and the hearts were examined macro- and microscopically within 48 h of their respective acceleration regimen. Eight transmural samples were taken from each heart for hydroxyproline analysis. Subendocardial hemorrhage (scale = 0-4) and myocyte damage (damaged cells per tissue section), respectively, were greatest in the control (2.08 and 0.87) and the 1-d (2.39 and 0.84) exposure groups, declining thereafter to the lowest values in the 6-month (0.33 and 0.06) group. Hydroxyproline concentration, as an indicator of possible scar-tissue development, showed a significant positive trend with increasing time at several sample sites on the left ventricular free wall. Heart rate (HR) and ventricular ectopy were highest during the early + Gz exposures and decreased with time thereafter. Seven additional swine were chronically instrumented with an arterial catheter for blood collection during the SACM to determine hematocrit and plasma levels of catecholamines, cortisol, and protein. The animals were exposed to the same 5 SACMs/d, 3 times/week for 4 months. Plasma norepinephrine (norepi), epinephrine (epi), and cortisol increased, respectively, from mean pre-G values of 0.9 ng/ml, 0.8 ng/ml, and 10.8 micrograms/dl to mean high values of 53.8 ng/ml, 58.1 ng/ml, and 34.0 micrograms/dl during 1-d and 1-week +Gz exposure; then declined to high values of 8.6 ng/ml, 4.3 ng/ml, and 18.8 micrograms/dl after 4 months of +Gz. The decline in macro- and microscopic myocardial damage during the approximate time frame of the decline in HR, arrhythmias, norepi, epi, and cortisol suggests a causative relationship. This decline also suggests that the injury observed during +Gz exposures is the result of the emotional stress of handling, restraint, and the unfamiliar environment of the centrifuge and +Gz, to which the animal becomes accustomed with time. However, the hydroxyproline and histologic data also suggest that myocardial scar-tissue may develop as a result of multiple +Gz exposures.

Acceleration↗

A twin registry study of familial and individual risk factors for trauma exposure and posttraumatic stress disorder.

This study examines the association of individual and familial risk factors with exposure to trauma and posttraumatic stress disorder (PTSD) in male twins (N = 6744) from the Vietnam Era Twin Registry. Independent reports of familial psychopathology from co-twins were used to avoid the potential biases of the family history method. Risk for exposure to traumatic events was increased by service in Southeast Asia, preexisting conduct disorder, preexisting substance dependence, and a family history of mood disorders whose effects appear to be partly genetic. Preexisting mood disorders in the individual were associated with decreased odds of traumatic exposure. Risk of developing PTSD following exposure was increased by an earlier age at first trauma, exposure to multiple traumas, paternal depression, less than high school education at entry into the military, service in Southeast Asia, and preexisting conduct disorder, panic disorder or generalized anxiety disorder, and major depression. Results suggest the association of familial psychopathology and PTSD may be mediated by increased risk of traumatic exposure and by preexisting psychopathology.

Adult↗

Copper inhibits the induction of stress protein synthesis by elevated temperatures in embryos of the sea urchin Strongylocentrus purpuratus.

A major component of the cellular stress response entails the induced synthesis of a suite of stress proteins under environmentally adverse conditions that functions to protect organisms from environmentally induced damage. Here, we examined induction of the stress response in the embryos of the sea urchin Strongylocentrus purpuratus under a combination of environmentally realistic conditions. First, we examined the response elicited over a range of free cupric ion activities, (Cu2+), using a metal buffer system to control trace metal speciation. We observed no pronounced differences in translational patterns in embryos exposed to free cupric ion activities, (Cu2+), of 10(-13)-10(-9) M by metabolic labeling, 1-dimensional electrophoresis and autoradiography. Further separation by 2-dimensional electrophoresis, however, revealed electrophoretically discernable variants of several groups of proteins at the higher Cu concentrations and the synthesis of a 60 kDa protein at (Cu2+) of 10(-9) M. In addition, there were differences in the stress response induced by heat-shock treatment in embryos cultured in seawater with different Cu concentrations; radiolabel was incorporated into a greater number of cellular proteins in embryos at lower (Cu2+) and the induced synthesis of stress proteins was greater. These data suggest that elevations in (Cu2+) impair the ability of the embryos to mount the stress response upon exposure to elevated temperatures and that Cu may alter critical developmental pathways by inhibiting the synthesis of regulatory proteins. Such effects on gene expression can result in manifestations that have been widely attributed to Cu toxicity, including developmental abnormalities and increased sensitivity to environmental extremes. We suggest that the particular sensitivity of embryonic systems upon exposure to multiple stressors may be a consequence of these mechanisms.

Animals↗

Cancer risk in laboratory workers: an emphasis on biological research.

BACKGROUND: At the end of the 1980s, several cancer clusters were observed in biological research laboratories. Over time, biological research and the technologies used have been diverse and have involved a wide range of mutagenic or carcinogenic chemical, physical and biological agents. METHODS: We reviewed 45 published studies on cancer risk among biological research personnel and workers in closely related fields, and numerous reports based on routinely collected data. RESULTS: Biological research could be associated with an elevated risk for pancreatic cancer, brain tumors, and certain hemopathies. A common limitation of available studies was low statistical power and the absence or inaccuracy of data on individual past exposure. CONCLUSIONS: Overall, this study suggests a low overall risk of cancer, albeit a higher risk may be suggested for cancers of the pancreas (risk ratios ranging from 0.5 to 6.3) and brain (0.7-9.4), and for non-Hodgkin's lymphoma (0.6-51.5). We suggest ways in which multiple past exposures could be assessed more precisely and emphasize a pressing need to take into account known confounders.

Biological Science Disciplines↗

Cystoisospora belli: in vitro multiplication in mammalian cells.

Intracellular development of Cystoisospora belli was demonstrated in 4 different mammalian cell lines. Human ileocecal adenocarcinoma (HCT-8), epithelial carcinoma of lung (A549), Madin-Darby bovine kidney (MDBK), and African green monkey kidney (VERO) were exposed in vitro to C. belli sporozoites, which had been isolated from the feces of HIV-AIDS patients. Parasites invaded all the cellular types between 4 and 12h after exposure and multiplication was demonstrated after 24 h. Grater number of merozoites formed in VERO cells, followed by HCT-8. In the MDBK and HCT-8 cells, the parasitophorous vacuole was less evident and immobile merozoites were observed in the cytoplasm. In VERO cells, one or several parasitophorous vacuoles contained up to 16 mobile sporozoites. No oocysts were found in any of the cell types used. VERO cells may be suitable for studies of the interaction between parasite and host cells.

AIDS-Related Opportunistic Infections↗

Whole-body biodistribution, radiation dosimetry estimates for the PET norepinephrine transporter probe (S,S)-[18F]FMeNER-D2 in non-human primates.

BACKGROUND: (S,S)-[F]FMeNER-D2 is a recently developed norepinephrine transporter ligand which is a potentially useful radiotracer for mapping the brain and heart norepinephrine transporter in vivo using positron emission tomography. In this work, we quantified the biodistribution over time and radiation exposure to multiple organs with (S,S)-[F]FMeNER-D2. METHODS: Whole-body images were acquired for 21 time points in two cynomolgus monkeys for approximately 270 min after injection of radioligand. Compressed 3-D to 2-D planar images were used to identify organs with the highest radiation exposure at each time point. Estimates of the absorbed dose of radiation were calculated using the MIRDOSE 3.1 software program performed with the dynamic bladder and ICRP 30 gastrointestinal tract models. RESULTS: In planar images, peak values of the percent injected dose (%ID) at a time after radioligand injection were calculated for the lungs (26.76% ID at 1.42 min), kidneys (13.55% ID at 2.18 min), whole brain (5.65% ID at 4.48 min), liver (7.20% ID at 2 min), red bone marrow (5.02% ID at 2.06 min), heart (2.36% ID at 1.42 min) and urinary bladder (23% ID at 250 min). Assuming a urine voiding interval of 2.4 h, the four organs with highest exposures in microGy . MBq ( mrad . mCi) were kidneys 126 (468), heart wall 108 (399), lungs 88.4 (327) and urinary bladder 114 (422). The effective doses were estimated with and without urine voiding at a range of 123 (33) and to 131 (35.5) microGy . MBq ( mrad . mCi). CONCLUSION: The estimated radiation burden of (S,S)-[F]FMeNER-D2 is comparable to that of other F radioligands.

Animals↗

A controlled comparison of symptoms and chemical intolerances reported by Gulf War veterans, implant recipients and persons with multiple chemical sensitivity.

Using the Environmental Exposure and Sensitivity Inventory (EESI), a standardized instrument for measuring chemical sensitivity, we obtained and compared ratings of symptoms, chemical (inhalant) intolerances, other intolerances (e.g., drugs, caffeine, alcohol, skin contactants), lifeimpact, and masking (ongoing exposures) in five populations: multiple chemical sensitivity (MCS) patients who did (n = 96) or did not (n = 90) attribute onset of their illness to a specific exposure event, patients with implanted devices (n = 87), Gulf War veterans (n = 72), and controls (n = 76). For each patient group, mean scores on the first four scales were significantly greater than for controls. MCS patients reported avoiding more chemical exposures (were less masked) than the other groups. Across groups, for a given level of symptoms, as masking increased, mean scores on the Chemical Intolerance Scale decreased. In contrast, mean scores on the Other Intolerance Scale appeared to be less affected by masking. These findings suggest that some patients with antecedent chemical exposures, whether exogenous (chemical spill, pesticide application, indoor air contaminants) or endogenous (implant), develop new chemical, food, and drug intolerances. Reports of new caffeine, alcohol, medication, food, or other intolerances by patients may signal exposure-related illness. Masking may reduce individuals' awareness of chemical intolerances, and, to a lesser degree, other intolerances.

Adult↗

Differential regional zif268 messenger RNA expression in an escalating dose/binge model of amphetamine-induced psychosis.

Amphetamine-induced psychosis is most often associated with a high-dose multiple binge pattern of stimulant abuse. To simulate these conditions in rats, we used an escalating dose/binge administration paradigm. Animals were pretreated with escalating doses of amphetamine (1.0-8.0mg/kg) over four days, then exposed to nine daily binges (8.0mg/kg every 2h; four injections/day). Other animals received either multiple injections of saline, saline followed by acute amphetamine (8.0mg/kg) or single daily injections of amphetamine (8.0mg/kg) in parallel with the escalating dose/binge treatment. One hour after the last injection, all animals were decapitated and regional brain activation patterns were assessed using in situ hybridization with antisense probes for zif268. Acute amphetamine resulted in a significant elevation of zif268 messenger RNA in both the nucleus accumbens and dorsal striatum. However, whereas after single daily amphetamine treatment this index was no longer elevated above control levels in the dorsal striatum, multiple binge exposures were required for the nucleus accumbens to return to baseline. Agranular insular cortex and medial olfactory tubercle zif268 messenger RNA expression was also markedly increased after acute amphetamine treatment but, unlike the nucleus accumbens and dorsal striatum, this increase was not significantly attenuated by either single daily injection or multiple binge treatment. Zif268 messenger RNA expression in the lateral nucleus of the amygdala also remained elevated above baseline after binge treatment. The possible relationships of these changes in zif268 messenger RNA regional expression patterns to the development of psychosis in high-dose stimulant abusers are discussed.

Amphetamines↗

Evaluation of oxidative stress and genotoxicity in organophosphorus insecticide formulators.

The aim of this study was to evaluate genotoxicity and oxidative stress in workers who formulate organophosphorus (OP) pesticides. In this survey, blood leukocytes and erythrocytes of a group of 21 pesticide formulating workers and an equal number of control subjects were examined for genotoxicity and oxidative stress parameters. The mean comet tail length and mean comet length were used to measure DNA damage. Lipid peroxidation level, catalase, superoxide dismutase (SOD) and glutathione peroxidase activities in erythrocytes were analysed as biomarkers of oxidative stress. In addition, the acetylcholinesterase activity was measured as a biomarker of toxicity. The average duration of employment of workers in the factory was 97 months. Results indicated that chronic exposure (multiple-dose, greater than or equal to 6 months duration) to OP pesticides was associated with increased activities of catalase, SOD and glutathione peroxidase in erythrocytes. The level of lipid peroxidation and acetylcholinesterase activity did not show any significant differences between the two groups. The results also indicated that chronic exposure to OP pesticides was associated with increased DNA damage. It is concluded that human chronic exposure to OP pesticides may result in stimulated antioxidant enzymes and increased DNA damage in the absence of depressed acetylcholinesterase levels. Routine genotoxicity monitoring concomitant to acetylcholinesterase activity in workers occupationally exposed to OP insecticides is suggested.

Adult↗

[Assessment of cumulative risk of mixtures with multiple target organs].

Humans are exposed to very complex environmental mixtures. In contrast to this reality, most laboratory investigations of mixtures, with exception of mutagenicity assays, have focused on mixtures comprised of a few chemicals. Component-based approaches that use information on the individual chemical contained in the mixture are not by themselves sufficient for complex, environmental mixtures because significant portions of the mixtures mass are unidentified. Assessment of the cumulative risk posed by exposure to multiple chemicals is a problem the USEPA's and FDA's program discuss regularly. In USA the Food Quality Protection Act of 1996 directs the Office of Pesticide Programs to include in its assessments the risk associated with the cumulative effects of pesticides that have a common mode of action. organophosphorus pesticides (OPs) have been the first class addressed, based on the common mechanism of acetylcholinesterase inhibition. Interestingly toxicity studies focussing on component-based analysis of mixtures indicated that for chemicals not sharing the same target and mechanism, the type of combined action or interaction found when each chemical in the mixture was present at clearly toxic effect levels did not predict the response observed when each chemical was present at or below the lowest observed adverse effect level (LOAEL). Moreover, mixtures of chemicals produced toxicity only for those chemicals that showed the same toxic effect on the same target organ when given singly at dose above the LOAEL values. However, end-points chosen in routine toxicity studies do not offer a wide dynamic range of testable, significant responses around and above the NOAEL. So far sensitive methods that detect low level changes have not yet been employed in mixtures testing protocol. Toxicogenomics integrated functional genomics with classical toxicology and we believe that it has great potential to revolutionize mixtures research.

Environmental Exposure↗

Multiple sclerosis unrelated to dog exposure.

We conducted a case-control study to evaluate the possible association between exposure to dogs and MS. Seventy cases were compared with 70 age- and sex-matched neighborhood controls and 57 cases with 57 age- and sex-matched clinic controls. No association was found, by age groups or by time periods before onset of MS, between MS and presence of any dog, a small dog, a medium or large dog, or an indoor dog in the household. There was a significant negative association between MS and presence of cats in the household and MS and presence of medium and large dogs in the household, and a significant positive association, for several age groups and time periods, between MS and a history of canine distemper in a household dog. The basis for these significant associations is not clear. This study adds weight to the evidence against an association between exposure to small or indoor dogs and MS.

Adolescent↗

Detection and differentiation of Xylella fastidiosa strains acquired and retained by glassy-winged sharpshooters (Hemiptera: Cicadellidae) using a mixture of strain-specific primer sets.

Xylella fastidiosa Wells is a bacterial pathogen that causes a variety of plant diseases, including Pierce's disease (PD) of grapevine, almond leaf scorch, alfalfa dwarf, citrus variegated chlorosis, and oleander leaf scorch (OLS). Numerous strains of this pathogen have been genetically characterized, and several different strains occur in the United States. The dominant vector in southern California is the glassy-winged sharpshooter, Homalodisca coagulata (Say) (Hemiptera: Cicadellidae). The high mobility of this insect, and its use of large numbers of host plant species, provides this vector with ample exposure to multiple strains of X. fastidiosa during its lifetime. To learn more about the ability of this vector to acquire, retain, and transmit multiple strains of the pathogen, we developed a polymerase chain reaction (PCR)-based method to detect and differentiate strains of X. fastidiosa present in individual glassy-winged sharpshooter adults. Insects were sequentially exposed to plants infected with a PD strain in grapevine and an OLS strain in oleander. After sequential exposure, a few insects tested positive for both strains (7%); however, in most cases individuals tested positive for only one strain (29% PD, 41% OLS). In transmission studies, individual adults transmitted either the PD or OLS strain of the pathogen at a rate (39%) similar to that previously reported after exposure to a single strain, but no single individual transmitted both strains of the pathogen. PD and OLS strains of X. fastidiosa remained detectable in glassy-winged sharpshooter, even when insects were fed on a plant species that was not a host of the strain for 1 wk.

Animals↗

Placental examination in intrauterine coinfection with herpes simplex virus and cytomegalovirus.

Intrauterine coinfections have rarely been reported. However, pregnancies exposed to multiple sexually transmitted infectious agents and drugs are likely to occur with increasing frequency and lead to complex pathology in the newborn. Often it will be difficult to establish a diagnosis, above all when this has to be done retrospectively. A premature (34 weeks) newborn presented with a complex clinical picture after exposure to multiple infectious and noninfectious teratogens during gestation. Immunocytochemical staining of the placental membranes and parenchyma suggested intrauterine coinfection by herpes simplex virus (HSV) type 2 and cytomegalovirus. This case illustrates the importance of careful placental investigation with modern techniques for the diagnosis of intrauterine HSV infection and coinfections.

Adolescent↗

Treatment of anxiety during pregnancy: effects of psychotropic drug treatment on the developing fetus.

Pregnancy is a time of great emotional change for a woman, producing increased stress and anxiety. Medication may be required for the treatment of anxiety disorders at this time. Given the fact that psychotropic drugs readily cross the placenta and could have important implications for the developing fetus, it is necessary to balance the possible effects of medication against the potential effects to both the mother and fetus if the anxiety disorder is left untreated. Despite the widespread use of psychotropic drugs such as benzodiazepines and antidepressants during pregnancy, there is a paucity of information regarding the effect of such exposure on the developing fetus. From a review of the literature it is clear that the issue of safety of psychotropic drugs during pregnancy is far from resolved. While some of the findings from animal studies are alarming, these studies cannot be directly extrapolated to humans. In addition, varying sample sizes and multiple drug exposures further complicate interpretation of human studies. Nonpharmacological treatments such as cognitive behavioural therapy should be employed whenever possible for the treatment of anxiety disorders during pregnancy. However, if medication is required pregnant women should be prescribed the lowest dosage for the minimum amount of time.

Abnormalities, Drug-Induced↗

Rapid virus dissemination in infant macaques after oral simian immunodeficiency virus exposure in the presence of local innate immune responses.

A vaccine to protect human immunodeficiency virus (HIV)-exposed infants is an important goal in the global fight against the HIV pandemic. Two major challenges in pediatric HIV vaccine design are the competence of the neonatal/infant immune system in comparison to the adult immune system and the frequent exposure to HIV via breast-feeding. Based on the hypothesis that an effective vaccine needs to elicit antiviral immune responses directly at the site of virus entry, the pattern of virus dissemination in relation to host immune responses was determined in mucosal and lymphoid tissues of infant macaques at 1 week after multiple oral exposures to simian immunodeficiency virus (SIV). The results show that SIV disseminates systemically by 1 week. Infant macaques can respond rapidly to virus challenge and mount strong innate immune responses. However, despite systemic infection, these responses are most pronounced in tissues close to the viral entry site, with the tonsil being the primary site of virus replication and induction of immune responses. Thus, distinct anatomic compartments are characterized by unique cytokine gene expression patterns. Importantly, the early response at mucosal entry sites is dominated by the induction of proinflammatory cytokines, while cytokines with direct antiviral activity, alpha/beta interferons, are only minimally induced. In contrast, both antiviral and proinflammatory cytokines are induced in lymphoid tissues. Thus, although infant macaques can respond quickly to oral viral challenge, the locally elicited immune responses at mucosal entry sites are likely to favor immune activation and thereby virus replication and are insufficient to limit virus replication and dissemination.

AIDS Vaccines↗

Long-term potentiation in the rat hippocampus is reversibly depressed by chronic intermittent ethanol exposure.

Alcohol exposure induces multiple neuroadaptive changes in the CNS that can have serious long-term consequences on CNS function including cognitive effects and attenuation of learning and memory. The cellular mechanisms underlying the CNS effects of alcohol have yet to be fully elucidated and are likely to depend on the pattern and dose of alcohol exposure. Using electrophysiological recordings from hippocampal slices obtained from control and chronic alcohol-treated rats, we have investigated the effects of a binge pattern of alcohol abuse on synaptic plasticity in the CNS. The alcohol-treated animals were exposed to ethanol vapor for 12-14 days using an intermittent exposure paradigm (14 h ethanol exposure/10 h ethanol withdrawal daily; blood alcohol levels approximately 180 mg/dl), a paradigm that models human binge alcohol use. Induction of long-term potentiation (LTP) in the CA1 region of the hippocampus by tetanic stimulation of Schaffer collaterals was completely blocked in slices from the chronic alcohol-treated animals. LTP remained blocked 1 day after withdrawal of animals from alcohol, indicating that the neuroadaptive changes produced by alcohol were not readily reversible. Partial recovery was observed after withdrawal from alcohol for 5 days. Other measures of synaptic plasticity including posttetanic potentiation and paired-pulse facilitation were also altered by the intermittent alcohol treatment paradigm. The results suggest that alterations in synaptic plasticity induced by chronic intermittent ethanol consumption play an important role in the effects of binge alcohol use on learning and memory function.

Alcoholism↗