Morphologically mixed chemical-electrical synapses formed by primary afferents in rodent vestibular nuclei as revealed by immunofluorescence detection of connexin36 and vesicular glutamate transporter-1.

Axon terminals forming mixed chemical/electrical synapses in the lateral vestibular nucleus of rat were described over 40 years ago. Because gap junctions formed by connexins are the morphological correlate of electrical synapses, and with demonstrations of widespread expression of the gap junction protein connexin36 (Cx36) in neurons, we investigated the distribution and cellular localization of electrical synapses in the adult and developing rodent vestibular nuclear complex, using immunofluorescence detection of Cx36 as a marker for these synapses. In addition, we examined Cx36 localization in relation to that of the nerve terminal marker vesicular glutamate transporter-1 (vglut-1). An abundance of immunolabeling for Cx36 in the form of Cx36-puncta was found in each of the four major vestibular nuclei of adult rat and mouse. Immunolabeling was associated with somata and initial dendrites of medium and large neurons, and was absent in vestibular nuclei of Cx36 knockout mice. Cx36-puncta were seen either dispersed or aggregated into clusters on the surface of neurons, and were never found to occur intracellularly. Nearly all Cx36-puncta were localized to large nerve terminals immunolabeled for vglut-1. These terminals and their associated Cx36-puncta were substantially depleted after labyrinthectomy. Developmentally, labeling for Cx36 was already present in the vestibular nuclei at postnatal day 5, where it was only partially co-localized with vglut-1, and did not become fully associated with vglut-1-positive terminals until postnatal day 20-25. The results show that vglut-1-positive primary afferent nerve terminals form mixed synapses throughout the vestibular nuclear complex, that the gap junction component of these synapses contains Cx36, that multiple Cx36-containing gap junctions are associated with individual vglut-1 terminals and that the development of these mixed synapses is protracted over several postnatal weeks.

Secondary poisoning of eagles following intentional poisoning of coyotes with anticholinesterase pesticides in western Canada.

Records of eagles, coyotes (Canis latrans), and red foxes (Vulpes vulpes) necropsied at the Western College of Veterinary Medicine, Saskatoon, Saskatchewan, Canada, between 1967 and 2002 were reviewed for cases suggestive of anticholinesterase poisoning. From 1993 to 2002, 54 putative poisoning incidents involving 70 bald eagles (Haliaeetus leucocephalus) and 10 golden eagles (Aquila chrysaetus) were identified. Of these, 50 incidents occurred in Saskatchewan, two were in Manitoba, and one occurred in each of Alberta and the Northwest Territories. The diagnosis was confirmed in eight instances by demonstration of pesticide in ingesta from eagles or known use of pesticide at the site together with brain cholinesterase (AChE) reduction of >50% in at least one animal. A presnmptive diagnosis of poisoning was made in 33 incidents based on brain AChE reduction of >50% in at least one animal; 13 incidents were considered suspicious because of circumstantial evidence of the death of eagles in association with other species and limited AChE reduction. Other wild species were found dead in 85% of the incidents involving eagles. Coyotes, foxes, black-billed magpies (Pica pica), and striped skunks (Mephitis mephitis) were associated with 34, six, six, and three incidents, respectively. There were eight additional incidents that did not involve eagles in which poisoning was diagnosed in coyotes. Carbofuran was identified in nine incidents. Carbamate poisoning was indicated on the basis of reactivation of brain AChE activity in two additional incidents. Brain AChE activity was not reduced from normal in eagles in four of seven incidents in which carbofuran was identified. The organophosplorous insecticide terbufos was found together with carbofuran in one incident. Brain AChE activity was measured in wild canids and in eagles in 15 incidents; in all of these incidents, brain AChE was redulced by >50% in at least one mammal, whereas this level of reduction occrred in eagles in only four incidents. Use of anticholinesterase pesticides to poison coyotes is illegal, but the practice continues and secondary poisoning of eagles is a problem of unknown proportions in western North America.

The effect of exposure to a commercial 2,4-D formulation during gestation on the immune response in CD-1 mice.

Pregnant CD-1 mice were administered a commercial 2,4-dichlorophenoxyacetic acid (2,4-D) formulation on days 6-16 days of gestation, in drinking water at concentrations ranging from 0 to 1.0% of the formulated product, equivalent to approximately 0-650 mg/kg per day expressed as the amine derivative. The effect of 2,4-D on immune function was evaluated in offspring 7 weeks after birth. The dams tolerated repeated 2,4-D exposure in drinking water without difficulty. The offspring exhibited decreased body weight with minor reductions in the kidney weights in the 0.1 and 1.0% 2,4-D treatment groups. A generalized suppression of lymphocyte stimulation by concanavalin A (Con A) was observed at high dose of commercial 2,4-D formulation (1.0%). Cytometric studies of the lymphocyte subpopulations demonstrated an increased relative count of B cells and reduced T cytotoxic or suppressor cells in the 1.0% formulation. The humoral immune response, antibody production against sheep red blood cells and peritoneal macrophage phagocytic function, were not altered by 2,4-D. Since the immune alterations in the offspring were observed many weeks after exposure, it appears as though 2,4-D exposure during gestation causes permanent changes in cell types associated with immune function. Since 2,4-D is not considered a persistent chemical, it is unlikely that 2,4-D residues are contributing significantly to the observed immune alterations. The immune alterations were observed only in the higher treatment groups. Therefore, the impact on human and animal health from an immune perspective, which would be encountered following normal application in the environment, would be minimal.

The meaning of the word "vertigo".

OBJECTIVE: To determine whether otologists see a need to further define the word "vertigo" and assess the variety of meanings that are meant in using the word. METHODS: A questionnaire was mailed to 720 members of the American Otological Society (AOS) and American Neurotological Society (ANS) to assess the need for further definition and the current understanding of the meaning of "vertigo." RESULTS: The response rate was 42%. Significant variability in the intended meaning of the word "vertigo" was found suggesting that the definition is inconsistent among experts. Three quarters of the respondents indicated that the word needs further definition. The word was most commonly used to indicate sensations that include spinning or turning only. CONCLUSION: The definition of the term "vertigo" is inconsistent among otolaryngologists. Refinement of the definition is desirable for accurate history taking and should be limited to false illusions of circular motion.

Suspect copper toxicity in an alpaca.

An alpaca presented in lateral recumbency and subsequently died. On necropsy examination the liver showed severe, widespread, periacinar hepatocellular necrosis, staining positive to a rhodamine stain for copper. Hepatic copper concentration was elevated. Copper toxicity in the camelid is difficult to diagnose, since the classical hemolytic crisis is not observed.

Local vestibular blood flow and systemic vascular responses to natural vestibular stimulation in the Mongolian gerbil.

HYPOTHESIS: Natural stimulation of the vestibular end organs will produce alterations in the local vestibular microvascular blood flow. BACKGROUND: The vestibular and cardiovascular systems require a coordinated interaction to maintain organ perfusion during rapid positional and postural changes. However, the detailed relationship of these systems is not well understood. There have been no previous descriptions of local vestibular blood flow (VBF) during natural stimulation (NS) conditions. METHODS: In vivo VBF and systemic blood pressure (BP) in the Mongolian gerbil during natural stimulation. Using laser Doppler flowmetry, the authors obtained continuous measures of local VBF in both anesthetized and alert gerbils during sinusoidal rotational stimuli. Simultaneous recordings of systemic BP were collected from the contralateral common carotid artery. RESULTS: The anesthetized gerbils showed stable VBF and BP during all vestibular stimuli. By contrast, alert subjects demonstrated a significant response to natural stimulation. The VBF increased 28% over baseline, and systemic BP increased 8% during a 45-second, 0.133-Hz sinusoid. Decreases in BP of 8% and 5%, respectively, were seen with a 0.10 and 0.20 Hz, 360-second stimulus. A corresponding determination of VBF during the extended stimulus conditions was not technically possible. CONCLUSIONS: To the authors' knowledge, these are the first in vivo descriptions of vestibular blood flow during natural stimulation. In the alert animals, VBF increased in response to NS. This increase in flow does not appear to be directly dependent on systemic blood pressure changes and indicates that the vestibular microvasculature is closely regulated.

Acoustic basis of tuning fork tests.

OBJECTIVES: To obtain pilot data to determine the feasibility of supporting or refuting a theory regarding lateralization of sound in conductive hearing loss and to describe the application of this theory to other tuning fork tests. DESIGN: Controlled audiometric testing with and without a conductive hearing loss. SETTING: A tertiary medical centre. METHODS: Sound pressure levels in external auditory canals were measured during presentation of 40- and 50-dB bone-conducted stimuli. Measurements were taken from five normal male subjects before and after inducing a conductive hearing loss. OUTCOME MEASURES: If sound intensity in the ear canal was greater in the unobstructed ear canal than the obstructed ear canal, the hypothesis was supported. The number of subjects required to provide definitive proof was calculated from the measured intensity difference with and without the conductive loss and the intrasubject variability of sound intensity measurements. The applicability of the theory to other tuning fork tests and auditory phenomena was explored. RESULTS AND CONCLUSIONS: A total of 600 subjects would be required to provide evidence to support our theory using this method. The acoustic impedance mismatch-reflected sound theory could be applied to other tuning fork tests, but until further proof is available, it must be considered only a theory.

The effect of exposure to a commercial 2,4-D herbicide formulation during gestation on urethan-induced lung adenoma formation in CD-1 mice.

Female CD-1 mice were exposed to a commercial amine formulation of 2,4-dichlorophenoxyacetic acid (2,4-D) on days 6-16 of gestation in drinking water at concentrations ranging from 0 to 1.0% of the formulated product, equivalent to approximately 0-650 mg/kg/d expressed as the amine derivative. The effect of 2,4-D on urethan-induced pulmonary adenoma formation was evaluated in female offspring 19 w after birth. Urethan-induced sleeping times observed following ip injection of 1.5 mg urethan/g bw 7 w after birth were not altered by 2,4-D (p = 0.10), indicating that 2,4-D did not affect the rate of urethan elimination. 2,4-D exposure did not affect the number of tumors produced (p = 0.58), but did reduce the mean tumor diameter in the highest dose group (p < 0.01). This minor antineoplastic activity of 2,4-D may be related, in part, to inhibitory effects of 2,4-D on various enzymatic or metabolic pathways, essential for cellular growth and tissue development. Since exposure to 2,4-D during pregnancy had little impact of tumor production, it is unlikely that persistent alteration to developing immune cells involved in the cell-mediated immunosurveillance mechanisms occurred. The subtle alteration in tumor size and the mild impairment of growth in the offspring were observed almost exclusively in the highest treatment group. Since this level of exposure is well in excess of those associated with normal application of 2,4-D, the hazard to non-target mammalian populations appears minimal.

Trace mineral and vitamin concentrations in the liver and serum of wild muskoxen from Victoria Island.

Selected trace minerals and vitamins were assayed in the liver and serum of 25 wild muskoxen (Ovibos moschatus) from Victoria Island, (Nunavut, Canada) in November, 1995. Mean +/- SE liver concentrations in micromol/kg wet weight were 260+/-16 for copper; 1.04+/-0.06 for selenium; 11.5+/-0.7 for molybdenum and 62.8+/-3.3 for vitamin E. Mean +/- SE serum concentrations in micromol/L were 14.2+/-0.3 for copper; 0.75+/-0.04 for selenium, 1.53+/-0.07 for vitamin A and 5.80+/-0.55 for vitamin E. Comparison of liver and serum concentrations of copper, selenium and vitamin E showed that the concentration in one tissue was a relatively poor indicator of the concentration in the other. The copper-molybdenum interaction often seen in domestic species was not observed. In general, the concentrations of metals and vitamins found in muskoxen were comparable to those in other ungulates although serum vitamin E concentrations were about one-fourth of those expected.

Suppression of rat and mouse lymphocyte function by urban air particulates (Ottawa dust) is reversed by N-acetylcysteine.

Epidemiology studies have demonstrated increased pulmonary morbidity such as allergy and infection with episodes of high particulate air pollution (size range 0.1-10 microm diameter, PM10), but the mechanism(s) for this association is not yet well defined. The present study was undertaken to evaluate the effects of EHC-93 urban particles (Ottawa dust) on immune functions of peripheral blood mononuclear cells (PBMCs) and splenocytes from male Fischer 344 rats and C57Bl/6 mice. Immune function endpoints evaluated included cell viability, lymphocyte blastogenesis stimulated by T-cell mitogen (concanavalin A, Con A) or B-cell mitogens [lipopolysaccharide (LPS) or LPS/dextran sulfate], intracellular Ca2+ concentration, interleukin 2 (IL-2) production, and expression of receptors for transferrin (TfR) and IL-2 (IL-2R). In addition, the effect of N-acetylcysteine (NAC), an antioxidant, on the toxicity of EHC-93 particles was evaluated. Total EHC-93 particles, water leachate of EHC-93, and washed EHC-93 suppressed proliferation of PBMCs and splenocytes to T- and B-cell mitogens. Treatment of splenocytes with EHC-93 particles did not alter intracellular Ca2+ concentration or mitogen-induced expression of TfR and IL-2R expression, but increased IL-2 production assayed by enzyme-linked immunosorbent assay (ELISA). In spite of an increase in IL-2 production, exogenous IL-2 when added to cultures was able to reverse the suppression of Con A-induced lymphocyte proliferation by EHC-93 particles. Furthermore, the suppressive effect of EHC-93 particles on mitogen-induced lymphocyte proliferation was completely abolished by addition of the antioxidant NAC to cultures, suggesting a possible role of oxidative factors for the toxicity of EHC-93 particles.

Flow cytometry as a tool to monitor the disturbance of phagocytosis in the clam Mya arenaria hemocytes following in vitro exposure to heavy metals.

The effectiveness of toxicology biomonitoring programs could be improved by the addition of sensitive biomarkers. In this study the cell viability and sensitivity of phagocytic function of phagocytes from bivalves (Mya arenaria) to selected heavy metals were measured by flow cytometry, a novel approach. Hemocytes (phagocytes) collected from bivalves by puncture of the posterior adductor muscle were incubated in vitro for 18 h in hemolymph containing 10(-9)-10(-3)M of cadmium chloride, zinc chloride, mercuric chloride, methylmercury chloride or silver nitrate, before determining their capacity to phagocytose fluorescent latex beads by flow cytometry. Heterogeneity of the hemocyte cell population was determined by forward scatter (FSC) and side scatter (SSC) cytometric profile which showed two distinct cell populations. At low doses (10(-9), 10(-8) M), all the metal compounds studied stimulated phagocytic activity except silver nitrate. At higher levels of exposure (10(-6), 10(7) M), all metals caused a significant concentration-related decrease in hemocyte phagocytosis activity. From the concentration of each metal inducing 50% suppression (IC50) of the phagocytic activity, the immunotoxic potential of metals with respect to phagocytic function can be ranked in the following increasing order: ZnCl2 < CdCl2 < AgNO3 < HgCl2 < CH3HgCl. Parallel analysis of hemocyte viability showed that suppression of phagocytosis by heavy metals was not solely related to a decreased cell viability. These results reveal the high but different degree of sensitivity of the phagocytosis activity of bivalves with respect to heavy metals, as measured by flow cytometry, and demonstrate that flow cytometry is a potentially useful tool in ecotoxicological monitoring.

Update on intratympanic gentamicin for Meniere's disease.

OBJECTIVE: To review the literature relating to intratympanic gentamicin injection therapy for Meniere's disease to detect consistencies and differences that might suggest optimal technique. DESIGN: Retrospective literature review. METHODS: Eighteen papers from the literature regarding clinical experience with intratympanic gentamicin injections for treatment of Meniere's disease were reviewed and tabulated. RESULTS: All papers reported high success rates in treating episodic vertigo of Meniere's disease, but technique, dose, duration, and treatment philosophy varied considerably. Hearing loss was typically reported in about 30% of patients. CONCLUSION: No single technique of gentamicin injection has a significant medical advantage over the others. Until controlled studies indicate otherwise, nonmedical needs such as convenience and safety may be considered when choosing a technique.

Toward a standard for electronystagmography.

Although the adoption of a standard is desirable, no person or laboratory will be entirely happy with whatever standard because ENG testing has become so individualized. Hopefully, a standard that will enhance patient care can be implemented without much cost.

Antidepressants and dizziness.

OBJECTIVE: To assess the clinical therapeutic significance of sleep disruption and admitted depression in patients with nonspecific dizziness. DESIGN: Retrospective database review. SETTING: Large tertiary care clinical practice. METHOD: Answers to questions about sleep disruption and depression from 137 patients who met inclusion criteria for nonspecific dizziness and a comparison group of 39 patients were contrasted. The Pearson chi-square test was used to determine the statistical significance of the results. MAIN OUTCOME MEASURE: Six weeks after beginning antidepressant therapy, patients were asked if their symptoms improved significantly. RESULTS: Patients with nonspecific dizziness reported more sleep disruption and admitted to depression more frequently than the comparison group. Persons with sleep disruption were more likely to respond to antidepressant therapy than those without sleep disruption. CONCLUSIONS: (1) Sleep disruption is common in persons with nonspecific dizziness. (2) Nonspecific dizziness may respond to antidepressant therapy, particularly in patients who report sleep disruption and/or depression.

Vestibular rehabilitation on a budget.

OBJECTIVE: This study was conducted to compare the results of an office-based vestibular rehabilitation program with the results from other larger programs in the literature. METHOD: Using a retrospective chart review, 138 dizzy patients seen in a tertiary medical centre were asked to perform directed exercises. Significant subjective improvement reported by the patients. RESULTS: Sixty-six percent of patients improved with this form of vestibular rehabilitation after 1 month. The results compare favourably with improvement rates from other centres in the literature with large, multidisciplinary teams. CONCLUSION: Vestibular rehabilitation therapy can be effectively administered from an office-based program.