Dog behaviours in veterinary consultations: Part II. The relationship between the behaviours of dogs and their owners.

Dogs synchronise their behaviour with those of their owners when confronted with an unfamiliar situation and interactions with their owners have been shown to decrease the dog's stress levels in some instances. However, whether owners may help manage dog anxiety during veterinary consultations remains unclear. In Part I, we compared the behaviour of dogs in the presence or absence of their owners during consultations, which consisted in three phases: exploration, examination, and greeting. Our findings suggest that allowing owners to attend consultations may be beneficial for dogs. In Part II, we investigated the direct relationship between owners' actions and their dog's behaviour. Using the videos from Part I, we examined whether: (1) dogs interact more when their owner is more interactive; (2) owners' stress scores are related to canine stress-related behaviour and emotional state; (3) owners' actions influence canine stress-related behaviours, emotional state and tolerance to manipulations; (4) canine stress-related behaviours and emotional state are associated with increased eye contact with their owners. We analysed the recordings of 29 dog-owner dyads submitted to a veterinary consultation in Part I. The behaviours of the dogs and their owners were analysed, and their emotional states were scored. The ease of manipulations was also scored. Despite limitations (e.g. no physical contact during examinations, no invasive procedures, aggressive dogs excluded, no male owners, limited sample size), our study showed a link between dog and owner behaviours: when owners attended an examination, their negative behaviours intensified the signs of anxiety in their dogs. Additionally, visual and verbal attempts to comfort their dog had no significant effect. However, we observed that the more dogs displayed stress-related behaviours, the more they established eye contact with their owners, suggesting that dogs seek information (through social referencing) or reassurance from their owners.

Dog behaviours in veterinary consultations: Part 1. Effect of the owner's presence or absence.

Veterinary practices can be stressful places for dogs. Decreasing stress during veterinary consultations is therefore a major concern, since animal welfare matters both for owners and veterinarians. Stress can be expressed through behaviour modifications; monitoring canine behaviour is thus one way to assess stress levels. We also know that the owner can affect dog behaviour in different ways. The aim of this study was therefore to assess the effect of the presence of owners on the behaviour of their dogs in veterinary consultations. We studied 25 dog-owner dyads at two standardised veterinary consultations, conducted at intervals of 5-7 weeks; the owner was present for the first consultation and absent for the second (O/NoO group, n = 12), or vice versa (NoO/O group, n = 13). A consultation consisted in three phases: exploration, examination, greeting. Dog behaviours were compared between the two conditions using a video recording. Despite some limitations (e.g. no male owners, the exclusion of aggressive dogs, a limited sample size, minimally invasive veterinary examinations, restricted owner-dog interactions), our results showed that the presence or absence of the owner had no significant effect on the stress-related behaviour of the dog or the veterinarian's ability to handle the animal during the examination phase (P > 0.05). Nevertheless, the behaviour of the dogs towards people was affected before, during, and after the veterinary examination. In the presence of their owner, dogs were more willing to enter the consultation room (P < 0.05), and they appeared more relaxed during the exploration phase (P < 0.01). During the examination, dogs looked in direction of their owner in both situations (owner present and behind the door, respectively; P < 0.001). These results suggest that allowing the owner to stay in the room during veterinary consultations is a better option for canine welfare.

Pseudoxanthoma elasticum overlaps hereditary spastic paraplegia type 56.

PURPOSE: Pseudoxanthoma elasticum (PXE) is a recessive disorder involving skin, eyes and arteries, mainly caused by ABCC6 pathogenic variants. However, almost one fifth of patients remain genetically unsolved despite extensive genetic screening of ABCC6, as illustrated in a large French PXE series of 220 cases. We searched for new PXE gene(s) to solve the ABCC6-negative patients. METHODS: First, family-based exome sequencing was performed, in one ABCC6-negative PXE patient with additional neurological features, and her relatives. CYP2U1, involved in hereditary spastic paraplegia type 56 (SPG56), was selected based on this complex phenotype, and the presence of two candidate variants. Second, CYP2U1 sequencing was performed in a retrospective series of 46 additional ABCC6-negative PXE probands. Third, six additional SPG56 patients were evaluated for PXE skin and eye phenotype. Additionally, plasma pyrophosphate dosage and functional analyses were performed in some of these patients. RESULTS: 6.4% of ABCC6-negative PXE patients (n = 3) harboured biallelic pathogenic variants in CYP2U1. PXE skin lesions with histological confirmation, eye lesions including maculopathy or angioid streaks, and various neurological symptoms were present. CYP2U1 missense variants were confirmed to impair protein function. Plasma pyrophosphate levels were normal. Two SPG56 patients (33%) presented some phenotypic overlap with PXE. CONCLUSION: CYP2U1 pathogenic variants are found in unsolved PXE patients with neurological findings, including spastic paraplegia, expanding the SPG56 phenotype and highlighting its overlap with PXE. The pathophysiology of ABCC6 and CYP2U1 should be explored to explain their respective role and potential interaction in ectopic mineralization.

Revisiting CFTR inhibition: a comparative study of CFTRinh -172 and GlyH-101 inhibitors.

BACKGROUND AND PURPOSE: For decades, inhibitors of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel have been used as tools to investigate the role and function of CFTR conductance in cystic fibrosis research. In the early 2000s, two new and potent inhibitors of CFTR, CFTRinh -172 and GlyH-101, were described and are now widely used to inhibit specifically CFTR. However, despite some evidence, the effects of both drugs on other types of Cl(-) -conductance have been overlooked. In this context, we explore the specificity and the cellular toxicity of both inhibitors in CFTR-expressing and non-CFTR-expressing cells. EXPERIMENTAL APPROACH: Using patch-clamp technique, we tested the effects of CFTRinh -172 and GlyH-101 inhibitors on three distinct types of Cl(-) currents: the CFTR-like conductance, the volume-sensitive outwardly rectifying Cl(-) conductance (VSORC) and finally the Ca(2+) -dependent Cl(-) conductance (CaCC). We also explored the effect of both inhibitors on cell viability using live/dead and cell proliferation assays in two different cell lines. KEY RESULTS: We confirmed that these two compounds were potent inhibitors of the CFTR-mediated Cl(-) conductance. However,GlyH-101 also inhibited the VSORC conductance and the CaCC at concentrations used to inhibit CFTR. The CFTRinh -172 did not affect the CaCC but did inhibit the VSORC, at concentrations higher than 5 µM. Neither inhibitor (20 µM; 24 h exposure) affected cell viability, but both were cytotoxic at higher concentrations. CONCLUSIONS AND IMPLICATIONS: Both inhibitors affected Cl(-) conductances apart from CFTR. Our results provided insights into their use in mouse models.

Role of CFTR in oxidative stress and suicidal death of renal cells during cisplatin-induced nephrotoxicity.

The clinical use of the antineoplastic drug cisplatin is limited by its deleterious nephrotoxic side effect. Cisplatin-induced nephrotoxicity is associated with an increase in oxidative stress, leading ultimately to renal cell death and irreversible kidney dysfunction. Oxidative stress could be modified by the cystic fibrosis transmembrane conductance regulator protein (CFTR), a Cl(-) channel not only involved in chloride secretion but as well in glutathione (GSH) transport. Thus, we tested whether the inhibition of CFTR could protect against cisplatin-induced nephrotoxicity. Using a renal proximal cell line, we show that the specific inhibitor of CFTR, CFTR(inh)-172, prevents cisplatin-induced cell death and apoptosis by modulating the intracellular reactive oxygen species balance and the intracellular GSH concentration. This CFTR(inh)-172-mediated protective effect occurs without affecting cellular cisplatin uptake or the formation of platinum-DNA adducts. The protective effect of CFTR(inh)-172 in cisplatin-induced nephrotoxicity was also investigated in a rat model. Five days after receiving a single cisplatin injection (5 mg/kg), rats exhibited renal failure, as evidenced by the alteration of biochemical and functional parameters. Pretreatment of rats with CFTR(inh)-172 (1 mg/kg) prior to cisplatin injection significantly prevented these deleterious cisplatin-induced nephrotoxic effects. Finally, we demonstrate that CFTR(inh)-172 does not impair cisplatin-induced cell death in the cisplatin-sensitive A549 cancer cell line. In conclusion, the use of a specific inhibitor of CFTR may represent a novel therapeutic approach in the prevention of nephrotoxic side effects during cisplatin treatment without affecting its antitumor efficacy.

Extracellular pH alkalinization by Cl-/HCO3- exchanger is crucial for TASK2 activation by hypotonic shock in proximal cell lines from mouse kidney.

We have previously shown that K(+)-selective TASK2 channels and swelling-activated Cl(-) currents are involved in a regulatory volume decrease (RVD; Barriere H, Belfodil R, Rubera I, Tauc M, Lesage F, Poujeol C, Guy N, Barhanin J, Poujeol P. J Gen Physiol 122: 177-190, 2003; Belfodil R, Barriere H, Rubera I, Tauc M, Poujeol C, Bidet M, Poujeol P. Am J Physiol Renal Physiol 284: F812-F828, 2003). The aim of this study was to determine the mechanism responsible for the activation of TASK2 channels during RVD in proximal cell lines from mouse kidney. For this purpose, the patch-clamp whole-cell technique was used to test the effect of pH and the buffering capacity of external bath on Cl(-) and K(+) currents during hypotonic shock. In the presence of a high buffer concentration (30 mM HEPES), the cells did not undergo RVD and did not develop outward K(+) currents (TASK2). Interestingly, the hypotonic shock reduced the cytosolic pH (pH(i)) and increased the external pH (pH(e)) in wild-type but not in cftr (-/-) cells. The inhibitory effect of DIDS suggests that the acidification of pH(i) and the alkalinization of pH(e) induced by hypotonicity in wild-type cells could be due to an exit of HCO(3)(-). In conclusion, these results indicate that Cl(-) influx will be the driving force for HCO(3)(-) exit through the activation of the Cl(-)/HCO(3)(-) exchanger. This efflux of HCO(3)(-) then alkalinizes pH(e), which in turn activates TASK2 channels.

Epidemiology of alveolar echinococcosis in southern Cantal, Auvergne region, France.

Alveolar echinococcosis (AE) is a helminth zoonosis which is encountered only in the northern hemisphere. In central France, the Auvergne region represents the most western and southern extension of this helminthiasis. In 1999, a human case of AE was diagnosed in the southern part of the Cantal department, where AE was supposed absent, and an epidemiological survey was subsequently carried out. The transmission of the zoonosis in the sylvatic and peridomestic definitive hosts was studied, as well as that in the rodent and human intermediate hosts. Eleven red foxes (Vulpes vulpes) were shot, and 50 fox faecal deposits were collected. Twelve farm dogs had their faeces taken by rectal touch, and four were checked after arecoline purgation. Optical detection of Echinococcus multilocularis worms was achieved on fox intestines after scraping, and also on dog stools after arecoline therapy. Coproantigen ELISA assay was performed for the 11 scraping products, for the 50 fox faeces, and for the 12 dog faecal samples. No adult AE agent was observed by microscopy, and the ELISA assay yielded positive results in one of 11 fox intestines, one of 50 fox faeces, and 2 of 12 dog faecal samples. Twenty-five small mammals were trapped, of which 19 were Arvicola terrestris water voles. One rodent liver exhibited a hepatic lesion consistent with AE. An epidemiological questionnaire was completed in 85 human volunteers, who were also serologically tested for AE. Only one (the case's husband) exhibited a Western-blotting pattern indicative of a low-grade AE infection. The results of this preliminary study suggested a slow AE extension to the south of Cantal department from the northern focus.

Inhibition of erythrocyte cation channels and apoptosis by ethylisopropylamiloride.

Even though lacking mitochondria and nuclei erythrocytes do undergo apoptotic cell death which is characterized by breakdown of phosphatidylserine asymmetry (leading to annexin binding), membrane blebbing and cell shrinkage. Previously, we have shown that erythrocyte apoptosis is triggered by osmotic shrinkage at least in part through activation of cell volume-sensitive cation channels and subsequent Ca2+ entry. The channels could not only be activated by cell shrinkage but as well by replacement of Cl- with gluconate. Both, channel activity and annexin binding were sensitive to high concentrations of amiloride (1 mM). The present study has been performed to search for more effective blockers. To this end channel activity has been evaluated utilizing whole-cell patch-clamp and annexin binding determined by FACS analysis as an indicator of erythrocyte apoptosis. It is shown that either, increase of osmolarity or replacement of Cl- by gluconate triggers the activation of the cation channel which is inhibited by amiloride at 1 mM but not at 100 microM. Surprisingly, the cation channel was significantly more sensitive to the amiloride analogue ethylisopropylamiloride (EIPA, IC(50)=0.6+/-0.1 microM, n=5). Exposure of the cells to osmotic shock by addition of sucrose (850 mOsm) led to stimulation of annexin binding which was inhibited similarly by EIPA (IC(50)=0.2+/-0.2 microM, n=4). Moreover, annexin binding was inhibited by higher concentrations of HOE 642 (IC(50)=10+/-5 microM, n=5) and HOE 694 (IC(50)=12+/-6 microM, n=4). It is concluded that osmotic shock stimulates a cation channel which participates in the triggering of erythrocyte apoptosis. EIPA is an effective inhibitor of this cation channel and of channel mediated triggering of erythrocyte apoptosis.

Cation channels trigger apoptotic death of erythrocytes.

Erythrocytes are devoid of mitochondria and nuclei and were considered unable to undergo apoptosis. As shown recently, however, the Ca(2+)-ionophore ionomycin triggers breakdown of phosphatidylserine asymmetry (leading to annexin binding), membrane blebbing and shrinkage of erythrocytes, features typical for apoptosis in nucleated cells. In the present study, the effects of osmotic shrinkage and oxidative stress, well-known triggers of apoptosis in nucleated cells, were studied. Exposure to 850 mOsm for 24 h, to tert-butyl-hydroperoxide (1 mM) for 15 min, or to glucose-free medium for 48 h, all elicit erythrocyte shrinkage and annexin binding, both sequelae being blunted by removal of extracellular Ca(2+) and mimicked by ionomycin (1 microM). Osmotic shrinkage and oxidative stress activate Ca(2+)-permeable cation channels and increase cytosolic Ca(2+) concentration. The channels are inhibited by amiloride (1 mM), which further blunts annexin binding following osmotic shock, oxidative stress and glucose depletion. In conclusion, osmotic and oxidative stress open Ca(2+)-permeable cation channels in erythrocytes, thus increasing cytosolic Ca(2+) activity and triggering erythrocyte apoptosis.

Time-dependent regulation of capacitative Ca2+ entry by IGF-1 in human embryonic kidney cells.

In a wide variety of cells, mitogenic factors release Ca(2+) from intracellular stores. The fall of the [Ca(2+)] within the lumen of the Ca(2+)-storing organelles triggers in many cells capacitative Ca(2+) entry (CCE). The present study was performed to elucidate the effect of insulin-like growth factor (IGF-1) on CCE in human embryonic kidney (HEK 293) cells. After depletion of Ca(2+) stores by thapsigargin, CCE was assessed by the increase in cytosolic free [Ca(2+)] (Fura-2 fluorescence imaging) when raising extracellular [Ca(2+)] from 0 to physiological concentrations. IGF-1 exposure (50 ng/ml) for 4 h in serum-free medium markedly enhanced CCE, while a 24-h exposure to IGF-1 depressed CCE profoundly. As some Ca(2+) channels are highly sensitive to the cell membrane potential, and as IGF-1 has been reported to enhance K(+) channel activity, the influence of K(+) channel blockers on the IGF-1-dependent stimulation of CCE was also tested. TEA, charybdotoxin and margatoxin decreased CCE. Similar to the total capacitative calcium entry, the fraction of CCE that was sensitive to K(+) channel blockers was increased after 4 h and decreased after 24 h exposure to IGF-1. Taken together, these data suggest that IGF-1 induces a transient increase followed by a decrease of CCE, and that these effects are at least partly dependent on IGF-1-induced K(+) channel activity.

Cerebral localization and regulation of the cell volume-sensitive serum- and glucocorticoid-dependent kinase SGK1.

The serum- and glucocorticoid-dependent kinase SGK1 is regulated by alterations of cell volume, whereby cell shrinkage increases and cell swelling decreases the transcription, expression and activity of SGK1. The kinase is expressed in all human tissues studied including the brain. The present study was performed to localize the sites of SGK1 transcription in the brain, to elucidate the influence of the hydration status on SGK1 transcription and to explore the functional significance of altered SGK1 expression. Northern blot analysis of human brain showed SGK1 to be expressed in all cerebral structures examined: amygdala, caudate nucleus, corpus callosum, hippocampus, substantia nigra, subthalamic nucleus and thalamus. In situ hybridization and immunohistochemistry in the rat revealed increased expression of SGK1 in neurons of the hippocampal area CA3 after dehydration, compared with similar slices from brains of euvolaemic rats. Additionally, several oligodendrocytes, a few microglial cells, but no astrocytes, were positive for SGK1. The abundance of SGK1 mRNA in the temporal lobe, including hippocampus, was increased by dehydration and SGK1 transcription in neuroblastoma cells was stimulated by an increase of extracellular osmolarity. Co-expression studies in Xenopus laevis oocytes revealed that SGK1 markedly increased the activity of the neuronal K+ channel Kv1.3. As activation of K+ channels modifies excitation of neuronal cells, SGK1 may participate in the regulation of neuronal excitability.

Genetic structure of Oestrus ovis populations in sheep and goats.

A genetic analysis using RAPD markers was performed on 12 natural populations of Oestrus ovis (Linné, 1761). Three-hundred and six O. ovis larvae (first, second and third instars) were randomly recovered in nasal cavities of sheep and goats naturally infected in Algeria, Ethiopia, France, Mauritania, Rumania and Tunisia and were analysed by 56 RAPD fragments. The results showed a high diversity within all samples. A significant genetic divergence was showed by discriminant analyses among the 12 populations sampled (p<0.0001). Moreover, discriminant analyses showed significant differentiation (p<0.0001) between O. ovis larva populations of sheep and goats and also among samples collected in the same region.

Efficacy of doramectin injectable against Oestrus ovis and gastrointestinal nematodes in sheep in the southwestern region of France.

A study was conducted to evaluate the therapeutic efficacy of doramectin administered intramuscularly at a dose rate of 200 microg/kg to sheep harbouring naturally acquired infections of gastrointestinal nematodes and Oestrus ovis in the southwestern region of France. On day 0, 24 sheep were selected on the basis of positive faecal egg counts (>100 EPG) and positive assessment of O. ovis infection (including positive O. ovis antibody level and positive clinical score). The sheep were randomly allocated to a non-medicated control group (T1) or a doramectin-treated group (T2) of 12 animals each. On day 0, sheep in group T2 received a single intramuscular injection of doramectin (200 microg/kg), whereas those in group T1 received an intramuscular injection of saline solution (sodium chloride, 0.02ml/kg). Individual faecal egg counts were performed on days 0, 1, 2, 3, 4, 5, 6, 7, and 14. Between days 14 and 16, all sheep were slaughtered, and worm and O. ovis burdens were determined. In doramectin-treated sheep, faecal egg counts had decreased to zero by day 4 for all recovered types of nematode eggs: strongyles, Nematodirus sp., Trichuris sp., and Rhabditidae sp. For strongyles, Nematodirus sp., and Rhabditidae, the percentage reductions in faecal egg counts (geometric means) of doramectin-treated sheep, compared to the non-medicated control sheep were 100% from days 4-7. For Trichuris sp., they were 100, 99.7, 99.9, and 100% on days 4, 5, 6, and 7, respectively. On day 14, percentage reductions were 100% for Nematodirus sp. and Rhabditidae, and 99.8 and 99.1% for strongyles and Trichuris sp., respectively. At necropsy, only adult nematodes and mainly first-stage O. ovis larvae were recovered. Doramectin was highly efficacious against the adult stages of Teladorsagia circumcincta (100%), Nematodirus battus (100%), Nematodirus filicollis (99.9%), Oesophagostomum venulosum (99.8%), and Trichuris sp. (99.3%). It was also 100% efficacious against first-stage larvae of O. ovis. No abnormal clinical signs or adverse reactions in any of the sheep treated with doramectin were observed.

Oestrus ovis (Diptera: Oestridae): effects of larval excretory/secretory products on nitric oxide production by murine RAW 264.7 macrophages.

Larvae of Oestrus ovis (Insecta: Diptera: Oestridae) are common parasites of nasal and sinus cavities of sheep and goats. Previous studies revealed that crude extracts of larvae modify NO synthesis by ovine monocyte derived macrophages. The aim of this study was to investigate the larval excretory/secretory products effects on nitric oxide production by murine tumour macrophages RAW 264.7. Stimulation of RAW macrophages by excretory/secretory products of the three instars larvae (25 microg/ml) significantly increased nitrite concentrations in culture supernatants compared to negative and positive Escherichia coli lipopolysaccharide control. This effect was time and dose dependent. Nitrite production in culture supernatants was due to induction of isoform NOS-2 because both NG monomethyl L-arginine (100 microM) and dexamethasone (20 microM) inhibited, by 60 and 50%, respectively, nitrite accumulation in culture supernatants. First steps of purification, by ion exchange chromatography, indicated that one protein of 29 kDa was able to induce NO synthesis by macrophages. Further studies are needed for a better characterization of these molecule and to investigate their immunogenicity for a vaccine approach.

Seasonal variation of Oestrus ovis-specific antibodies in sheep and goats mixed flocks in Greece.

The aim of this survey was to investigate the year-round epidemiological patterns of Oestrus ovis ELISA sero-prevalence in sheep and goats kept together under the same husbandry system in an endemic area of Greece. Twenty-five adult female sheep and 25 adult female goats, coming from a large mixed flock, were randomly selected, eartaged and monthly blood sampled during 1 year period (November 1998-October 1999). Serological prevalence in sheep was 100% all around the year. Mean intensities of specific O. ovis antibodies follow a seasonal evolution with higher mean titers between March and July than in winter. In contrast, the serological prevalences in goats were low specially in winter months (from October to January). No significant difference were noticed in goats antibody levels during the year period. The possible reasons of this difference of O. ovis sero-prevalence between sheep and goats are discussed.

Immunisation of lambs with excretory secretory products of Oestrus ovis third instar larvae and subsequent experimental challenge.

Excretory-secretory products (ESP) of myiasis producing agents are involved in nutrition and development of larvae and are often immunogens. This study was carried out in order to define the antigenicity, the immunogenicity of Oestrus ovis ESP and the role of sheep immune response to ESP. Twenty-four six to eight month old female lambs were randomly allocated into two groups. The first one was immunised twice, four weeks apart, with excretory-secretory products of Oestrus ovis third instar larvae (L3ESP) in complete then incomplete Freund adjuvant. The second one served as a control, and received two injections of PBS plus complete and incomplete Freund adjuvant. Fifteen and twenty-eight days after the second immunisation, animals of both groups were experimentally challenged with O. ovis first instar larvae. Twelve days after the second experimental challenge, the twenty-four lambs were necropsied. The total number of O. ovis larvae, their stages of development, weights and sizes were recorded per animal and compared between the two groups. Establishment rates were very similar in both groups: 39% and 35% in control and vaccinated groups respectively but the percentage of developing stages was higher in the control group (13%) than in the vaccinated group (6%). It was concluded that the L3ESP immunisation of sheep did not protect against larval establishment but provided an inhibitory effect on larval growth.

Potassium channels in primary cultures of seawater fish gill cells. I. Stretch-activated K(+) channels.

Previous studies using the patch-clamp technique demonstrated the presence of a small conductance Cl(-) channel in the apical membrane of respiratory gill cells in primary culture originating from sea bass Dicentrarchus labrax. We used the same technique here to characterize potassium channels in this model. A K(+) channel of 123 +/- 3 pS was identified in the cell-attached configuration with 140 mM KCl in the bath and in the pipette. The activity of the channel declined rapidly with time and could be restored by the application of a negative pressure to the pipette (suction) or by substitution of the bath solution with a hypotonic solution (cell swelling). In the excised patch inside-out configuration, ionic substitution demonstrated a high selectivity of this channel for K(+) over Na(+) and Ca(2+). The mechanosensitivity of this channel to membrane stretching via suction was also observed in this configuration. Pharmacological studies demonstrated that this channel was inhibited by barium (5 mM), quinidine (500 microM), and gadolinium (500 microM). Channel activity decreased when cytoplasmic pH was decreased from 7.7 to 6.8. The effect of membrane distension by suction and exposure to hypotonic solutions on K(+) channel activity is consistent with the hypothesis that stretch-activated K(+) channels could mediate an increase in K(+) conductance during cell swelling.

Potassium channels in primary cultures of seawater fish gill cells. II. Channel activation by hypotonic shock.

Previous studies performed on apical membranes of seawater fish gills in primary culture have demonstrated the existence of stretch-activated K(+) channels with a conductance of 122 pS. The present report examines the involvement of K(+) channels in ion transport mechanisms and cell swelling. In the whole cell patch-clamp configuration, K(+) currents were produced by exposing cells to a hypotonic solution or to 1 microM ionomycin. These K(+) currents were inhibited by the addition of quinidine and charybdotoxin to the bath solution. Isotopic efflux measurements were performed on cells grown on permeable supports using (86)Rb(+) as a tracer to indicate potassium movements. Apical and basolateral membrane (86)Rb effluxes were stimulated by the exposure of cells to a hypotonic medium. During the hypotonic shock, the stimulation of (86)Rb efflux on the apical side of the monolayer was inhibited by 500 microM quinidine or 100 microM gadolinium but was insensitive to scorpion venom [Leirus quinquestriatus hebraeus (LQH)]. An increased (86)Rb efflux across the basolateral membrane was also reduced by the addition of quinidine and LQH venom but was not modified by gadolinium. Moreover, basolateral and apical membrane (86)Rb effluxes were not modified by bumetanide or thapsigargin. There is convincing evidence for two different populations of K(+) channels activated by hypotonic shock. These populations can be separated according to their cellular localization (apical or basolateral membrane) and as a function of their kinetic behavior and pharmacology.

Prevalence and larval burden of oestrus ovis (Linné 1761) in sheep and goats in northern mediterranean region of France.

A slaughterhouse survey to determine prevalence and larval burden of Oestrus ovis larvae in sheep and goats was performed monthly during one year in Pézenas, South of France, northern mediterranean region. A total of 1303 sheep and goat heads were selected at random. O. ovis larvae were found in 274 sheep out of 631 (43.4%), and the prevalence rate varied from 14.3% in February to 65% in October. The mean number of larvae in infected sheep heads was 10.86 with 9.24 L1, 0.91 L2 and 0.71 L3. One hundred and ninety-one goats out of 672 were infected (28.4%), and the prevalence rate varied from 6.25% in September to 47.1% in April. In infected goat heads, the mean parasitic burden was 5.35 with 4.04 L1, 0.73 L2 and 0.58 L3. These results confirm worldwide observations indicating that the prevalence and the parasitic burdens are less in goats than in sheep.

Changing reactivity of caprine and ovine mononuclear phagocytes throughout part of the life cycle of Oestrus ovis: assessment through spontaneous and inductible NO production.

The in vitro reactivity of monocyte-derived macrophages (MDM) from Oestrus ovis (O. ovis) artificially infested lambs and kids was determined by measuring their production of nitric oxide (NO) during the course of infestation. In both species, crude antigenic preparations obtained from O. ovis first instar larvae (L1) were found to significantly (P < 0.01) inhibit this NO production, whereas O. ovis second instar (L2) extract stimulated it. Furthermore, this NO production by MDM decreased during infestation and was related to blood eosinophilia. It appears that crude antigenic extract from O. ovis modified the NO activity of macrophages from lambs and kids infested with O. ovis larvae.