Pharmacologic interaction between oxfendazole and triclabendazole: In vitro biotransformation and systemic exposure in sheep.

The aim of the current work was to evaluate a potential pharmacokinetic interaction between the flukicide triclabendazole (TCBZ) and the broad-spectrum benzimidazole (BZD) anthelmintic oxfendazole (OFZ) in sheep. To this end, both an in vitro assay in microsomal fractions and an in vivo trial in lambs parasitized with Haemonchus contortus resistant to OFZ and its reduced derivative fenbendazole (FBZ) were carried out. Sheep microsomal fractions were incubated together with OFZ, FBZ, TCBZ, or a combination of either FBZ and TCBZ or OFZ and TCBZ. OFZ production was significantly diminished upon coincubation of FBZ and TCBZ, whereas neither FBZ nor OFZ affected the S-oxidation of TCBZ towards its sulfoxide and sulfone metabolites. For the in vivo trial, lambs were treated with OFZ (Vermox® oral drench at a single dose of 5 mg/kg PO), TCBZ (Fasinex® oral drench at a single dose of 12 mg/kg PO) or both compounds at a single dose of 5 (Vermox®) and 12 mg/kg (Fasinex®) PO. Blood samples were taken to quantify drug and metabolite concentrations, and pharmacokinetic parameters were calculated by means of non-compartmental analysis. Results showed that the pharmacokinetic parameters of active molecules and metabolites were not significantly altered upon coadministration. The sole exception was the increase in the mean residence time (MRT) of OFZ and FBZ sulfone upon coadministration, with no significant changes in the remaining pharmacokinetic parameters. This research is a further contribution to the study of metabolic drug-drug interactions that may affect anthelmintic efficacies in ruminants.

Pharmacokinetic assessment of the monepantel plus oxfendazole combined administration in dairy cows.

Monepantel (MNP) is a novel anthelmintic compound launched into the veterinary pharmaceutical market. MNP is not licenced for use in dairy animals due to the prolonged elimination of its metabolite monepantel sulphone (MNPSO2 ) into milk. The goal of this study was to evaluate the presence of potential in vivo drug-drug interactions affecting the pattern of milk excretion after the coadministration of the anthelmintics MNP and oxfendazole (OFZ) to lactating dairy cows. The concentrations of both parent drugs and their metabolites were measured in plasma and milk samples by HPLC. MNPSO2 was the main metabolite recovered from plasma and milk after oral administration of MNP. A high distribution of MNPSO2 into milk was observed. The milk-to-plasma ratio (M/P ratio) for this metabolite was equal to 6.75. Conversely, the M/P ratio of OFZ was 1.26. Plasma concentration profiles of MNP and MNPSO2 were not modified in the presence of OFZ. The pattern of MNPSO2 excretion into milk was also unchanged in animals receiving MNP plus OFZ. The percentage of the total administered dose recovered from milk was 0.09 ± 0.04% (MNP) and 2.79 ± 1.54% (MNPSO2 ) after the administration of MNP alone and 0.06 ± 0.04% (MNP) and 2.34 ± 1.38% (MNPSO2 ) after the combined treatment. The presence of MNP did not alter the plasma and milk disposition kinetics of OFZ. The concentrations of the metabolite fenbendazole sulphone tended to be slightly higher in the coadministered group. Although from a pharmacodynamic point of view the coadministration of MNP and OFZ may be a useful tool, the presence of OFZ did not modify the in vivo pharmacokinetic behaviour of MNP and therefore did not result in reduced milk concentrations of MNPSO2 .

Assessment of the pharmacological interactions between the nematodicidal fenbendazole and the flukicidal triclabendazole: In vitro studies with bovine liver microsomes and slices.

Parasitic diseases have a significant impact on livestock production. Nematodicidal drugs, such as fenbendazole (FBZ) or its oxidized metabolite oxfendazole (OFZ), can be used along with the trematodicidal triclabendazole (TCBZ), to broaden the spectrum of anthelmintic activity. However, co-exposure to these compounds could lead to drug-drug (D-D) interactions and eventually alter the clinical profile of each active principle. The aim of this study was to assess the presence of such interactions by means of two in vitro models, namely bovine liver microsomal fractions and bovine precision-cut liver slices (PCLSs). To this end, an in vitro assessment involving incubation of FBZ and TCBZ or a combination of FBZ and TCBZ was carried out. Results with microsomal fractions showed a 78.4% reduction (p = .002) in the rate of OFZ production upon co-incubation, whereas the sulfoxide metabolite of TCBZ (TCBZSO) exhibited a decreasing tendency. With PCLS, OFZ accumulation in the incubation medium increased 1.8-fold upon co-incubation, whereas TCBZSO accumulation decreased by 28%. The accumulation of FBZ and OFZ in the liver tissue increased upon 2-hr co-incubation, from 2.1 ± 1.5 to 18.2 ± 6.1 (p = .0009) and from 0.4 ± 0.1 to 1.3 ± 0.3 nmol (p = .0005), respectively. These results confirm the presence of D-D interactions between FBZ and TCBZ. Further studies are needed to determine the extent of involvement of drug-metabolizing enzymes and membrane transporters in interactions between compounds largely used in livestock production systems.

Hepatic biotransformation pathways and ruminal metabolic stability of the novel anthelmintic monepantel in sheep and cattle.

Monepantel (MNP) is a new amino-acetonitrile derivative anthelmintic drug used for the treatment of gastrointestinal (GI) nematodes in sheep. The present work investigated the main enzymatic pathways involved in the hepatic biotransformation of MNP in sheep and cattle. The metabolic stability in ruminal fluid of both the parent drug and its main metabolite (monepantel sulphone, MNPSO2 ) was characterized as well. Additionally, the relative distribution of both anthelmintic molecules between the fluid and particulate phases of the ruminal content was studied. Liver microsomal fractions from six (6) rams and five (5) steers were incubated with a 40 µm of MNP. Heat pretreatment (50 °C for 2 min) of liver microsomes was performed for inactivation of the flavin-monooxygenase (FMO) system. Additionally, MNP was incubated in the presence of 4, 40, and 80 µm of methimazole (MTZ), a FMO inhibitor, or equimolar concentrations of piperonyl butoxide (PBx), a well-known general cytochrome P450 (CYP) inhibitor. In both ruminant species, MNPSO2 was the main metabolite detected after MNP incubation with liver microsomes. The conversion rate of MNP into MNPSO2 was fivefold higher (P < 0.05) in sheep (0.15 ± 0.08 nmol/min·mg) compared to cattle. In sheep, the relative involvement of both FMO and CYP systems (FMO/CYP) was 36/64. Virtually, only the CYP system appeared to be involved in the production of MNPSO2 in cattle liver. Methimazole significantly reduced (41 to 79%) the rate of MNPSO2 production in sheep liver microsomes whereas it did not inhibit MNP oxidation in cattle liver microsomes. On the other hand, PBx inhibited the production of MNPSO2 in liver microsomes of both sheep (58 to 98%, in a dose-dependent manner) and cattle (almost 100%, independently of the PBx concentration added). The incubation of MNP and MNPSO2 with ruminal contents of both species showed a high chemical stability without evident metabolism and/or degradation as well as an extensive degree of adsorption (83% to 90%) to the solid phase of the ruminal content. Overall, these results are a further contribution to the understanding of the metabolic fate of this anthelmintic drug in ruminants.

Intestinal drug transport: ex vivo evaluation of the interactions between ABC transporters and anthelmintic molecules.

The family of ATP-binding cassette (ABC) transporters is composed of several transmembrane proteins that are involved in the efflux of a large number of drugs including ivermectin, a macrocyclic lactone (ML) endectocide, widely used in human and livestock antiparasitic therapy. The aim of the work reported here was to assess the interaction between three different anthelmintic drugs with substrates of the P-glycoprotein (P-gp) and the breast cancer resistance protein (BCRP). The ability of ivermectin (IVM), moxidectin (MOX) and closantel (CST) to modulate the intestinal transport of both rhodamine 123 (Rho 123), a P-gp substrate, and danofloxacin (DFX), a BCRP substrate, across rat ileum was studied by performing the Ussing chamber technique. Compared to the controls, Rho 123 efflux was significantly reduced by IVM (69%), CST (51%) and the positive control PSC833 (65%), whereas no significant differences were observed in the presence of MOX (30%). In addition, DFX efflux was reduced between 59% and 72% by all the assayed drug molecules, showing a higher potency than that observed in the presence of the specific BCRP inhibitor pantoprazole (PTZ) (52%). An ex vivo intestinal transport approach based on the diffusion chambers technique may offer a complementary tool to study potential drug interactions with efflux transporters such as P-gp and BCRP.

Combined moxidectin-levamisole treatment against multidrug-resistant gastrointestinal nematodes: A four-year efficacy monitoring in lambs.

Nematicide combinations may be a valid strategy to achieve effective nematode control in the presence of drug resistance. The goal of the current trial was to evaluate the pharmaco-parasitological performance of the moxidectin (MOX) and levamisole (LEV) combination after four years of continuous use in lambs naturally parasitized with multi-resistant gastrointestinal nematodes. At the beginning of the trial, 40 lambs were divided into four groups (n = 10), which were untreated (control) or subcutaneously treated with MOX (0.2 mg/kg), LEV (8 mg/kg) or with the combination MOX + LEV (administered separately at 0.2 and 8 mg/kg, respectively). Blood samples were collected at different times post-treatment and LEV and MOX plasma concentrations were measured by HPLC. The clinical efficacy of the continuous use of MOX + LEV combination was assessed with the controlled efficacy test (CET), performed at the beginning and end of the study, and with the faecal egg count reduction (FECR) test, performed over the four-year study period. No significant adverse pharmacokinetic changes were observed either for MOX or LEV after their co-administration to infected lambs. The CET (first year) showed efficacies of 84.3 % (Haemonchus contortus), 100 % (Teladorsagia circumcincta and Trichostrongylus axei), and 97.4 % (T. colubriformis). After the repetitive use of the combined treatment for four years, those efficacies remained high (100 %) and only decreased to 58 % against T. colubriformis. The evaluation of the FECR over the study period showed fluctuations in the performance of the combined administration. The initial FECR (2014) was 99 % (MOX), 85 % (LEV) and 100 % (MOX + LEV). The co-administration of MOX + LEV during the four-year experimental period resulted in a significantly higher anthelmintic effect (87 %) than that of MOX (42 %) or LEV (69 %) given alone. The combined use of MOX + LEV to control resistant gastrointestinal nematodes appears to be a valid strategy under specific management conditions. A high initial therapeutic response to the combination would be a relevant feature for the success of this tool.

Are topical oestrogens useful in faecal incontinence? Double-blind randomized trial.

BACKGROUND: Histopathological studies have shown the presence of oestrogenic receptors in the anal sphincter, which presumes a role in muscular trophism for circulating oestrogens. This could explain the increase in faecal incontinence (FI) in postmenopausal women. OBJECTIVE: To evaluate the effect of topical oestrogens (TE) in controlling symptoms of FI in postmenopausal women. Hypothesis The application of TE in postmenopausal women with FI improves continence. METHOD: We performed a prospective double-blind randomized trial applying TE to the anal mucosa in postmenopausal women with FI. STUDY PERIOD: 2005-2006. The patients were randomized into two groups: I--topical estriol, II--placebo. In both groups, the ointment was applied three times daily for a period of 6 weeks. We compared Wexner's FI score and the FI quality of life scale, before commencing and after 6 weeks of application. RESULTS: In this period we evaluated 36 patients. Average age: 67 years (48-84). Group I: 18 patients and group II: 18 patients, one patient was excluded. Wexner's FI score in group I was 11 (5-18) and 7 (0-19) with pre- and postapplication respectively (P = 0.002). Wexner's FI score in group II was 12 and 9 with pre- and postapplication respectively (P = 0.013). When we compared the results between both groups, this was not statistically significant (P = 0.521). CONCLUSION: There is improvement of continence in both groups that had the ointment applied; nonetheless this study could not show that TE improves FI more than a placebo does.

Effects of fenbendazole and triclabendazole on the expression of cytochrome P450 1A and flavin-monooxygenase isozymes in bovine precision-cut liver slices.

Combinations of the anthelmintics fenbendazole (FBZ) and triclabendazole (TCBZ) have shown enhanced efficacy against the liver fluke Fasciola hepatica. This study aimed to measuring the constitutive expression of CYP1A1, CYP1A2, FMO1 and FMO3, thought to be involved in the metabolism of those compounds, by using an absolute quantitative real time (RT)-PCR approach in bovine precision-cut liver slices (PCLS). It also aimed to characterize the effects of FBZ and TCBZ (alone and in combination) on the expression and activity of the aforementioned isozymes. Both FMO1 and FMO3 were equally represented in control PCLS, whereas CYP1A2 was expressed more than CYP1A1 (P<0.05). PCLS cultured in the presence of beta naphthoflavone (ß-NF; CYP1A inducer) had higher mRNA levels of CYP1A1, CYP1A2, FMO1 and FMO3 (P<0.05). No clear-cut evidence of transcriptional effects of the anthelmintics were recorded. After incubation of PCLS with FBZ, there was a significant increase (P<0.05) vs. controls and TBCZ was observed for CYP1A1. PCLS treated with FBZ showed a higher (P<0.05) expression of CYP1A2 compared to controls, TCBZ alone, and the combination FBZ+TCBZ. The gene expression profiles of FMO1 and FMO3 were not affected by the presence of the anthelmintics; the only exception was an upregulation of FMO3 by TCBZ alone. The observed transcriptional effects of the xenobiotics were not mirrored by increased enzyme activities using prototypical substrates of the isozymes under study. Although further confirmatory studies are needed, these results suggest that PCLS represent an alternative in vitro tool for studies on the expression, regulation and function of relevant xenobiotic-metabolizing enzymes in cattle.

Exploring language barriers to Evidence-based Health Care (EBHC) in post-graduate medical students: a randomised trial.

BACKGROUND: Understanding the written English language might be a barrier when teaching Evidence-based Health Care (EBHC) to Spanish-speaking physicians. AIM: To quantify the magnitude of this potential barrier. METHOD: Cochrane Review abstracts in English or in Spanish were randomly distributed among first-year residents at the Pontificia Universidad Catolica of Chile. Residents answered investigator-designed questionnaires to measure their comprehension while the time needed to complete the task was recorded. RESULTS: Groups were similar at baseline. Mean score for those reading in Spanish was 11.9 +/- 2.8 (range 5 to 18) compared to 10.5 +/- 3.8 (range 1 to 17) for those reading in English (p=0.04). Low scores ( pound 9) were twice as frequent for the English group than for the Spanish group (16.7% vs 34.7%; p=0.042). The time to complete the task was also longer for the group reading in English. CONCLUSION: Language should be taken into account when teaching EBHC to Spanish-speaking physicians.

Factors influencing the success of aerial rabies vaccination of foxes.

Sylvatic rabies has been eradicated from most of Central Europe, but cases still occur in the Balkans. Oral rabies vaccination of foxes is an effective method for controlling the disease. The aim of this study was to evaluate the success of aerial vaccination campaigns conducted in Montenegro by identifying ecological, environmental and climatic factors that influenced the prevalence of antibodies to the rabies vaccine. To monitor the bait uptake and the serological responses to vaccination, foxes were shot by hunters. Of 175 shot foxes, 142 foxes (81.1%) had consumed baits. Of these only a total of 81 (57.0%) tested positive for rabies vaccine antibodies, possibly, due to the delayed uptake of bait in which the rabies vaccine was already inactivated. We found that low vaccination responses were associated with high fox density and bait delivery in open areas. In high fox density habitat, bait uptake might be delayed as other food and prey options for foxes are abundant. Similarly, delayed bait uptake probably occurred in open areas as such areas are less frequently used by foxes. The findings of this study suggest that efficacy of oral rabies vaccination by aerial delivery is associated with landscape features.

The relation between linear extent and velocity in drawing movements.

The speed of execution of complex movements depends on both the local, differential properties of the trajectory and on some of its more global metric parameters. The effects of these global factors were studied in free, writing-like movements with either piece-wise constant, or regularly changing curvature. It is demonstrated that the tangential velocity of the pen's tip is tightly correlated, through a power function, with the total linear extent of the trajectory (perimeter). Thus, a strong tendency exists to keep the execution time of these complex trajectories independent of the movement size (isochrony). Furthermore, it is shown that the average tangential velocity over identifiable segments of the trajectory also depends on the corresponding average curvature. The implications of these results vis-à-vis the central representation and planning of movements are discussed.

Trajectory determines movement dynamics.

The relation between figural and kimematic aspects of movement was studied in handwriting and drawing. It was found that, throughout the movement, the tangential velocity. V is proportional to the radius of curvature r of the trajectory: V= kr, or, equivalently, that the angular velocity is constant: dalpha(t)/dt = K. However, the constant k generally takes several distinct values during the movement, the changes being abrupt. These changes suggest a clear segmentation of the movement into units of action which overlap but do not coincide with the figural units as defined by the discontinuities of the movement (cuspids, points of inflection). This organisational principle holds even when movements are mechanically constrained or are executed under strict visuo-motor guidance. Moreover, the segmentation of a given trajectory is invariant with respect to the total duration of the movement. A tentative interpretation of the principle is proposed which results from the assumption that the actual movement is produced as a continuous approximation to an intended movement, and that the well known relationship between movement speed and extent in rectilinear trajectories (Fitts' law) also applies to such continuous approximation.

Internally driven vs. externally cued movement selection: a study on the timing of brain activity.

Brain imaging studies in man and single cell recordings in monkey have suggested that medial supplementary motor areas (SMA) and lateral pre-motor areas (PMA) are functionally dissociated concerning their involvement in internally driven and externally cued movements. This dichotomy, however, seems to be relative rather than absolute. Here, we searched for further evidence of relative differences and aimed to determine by what aspect of brain activity (duration, strength, or both) these might be accounted for. Event-related potentials (ERPs) were recorded while healthy, right-handed subjects selected one of three possible right hand digit movements based either on 'internal' choice or 'external' cues. The results obtained from ERP mapping suggest that movement selection evokes the same electrical brain activity patterns in terms of surface potential configurations in the same order and at the same strength independent of the selection mode. These identical configurations, however, differed in their duration. Combined with the results of a distributed source localization procedure, our data are suggestive of longer lasting activity in SMA during the 'internal' and longer lasting activity in PMA during the 'external' condition. Our results confirm previous findings in showing that SMA and PMA are distinctively involved in the two tasks and that this functional dichotomy is relative rather than absolute but indicate that such a dissociation can result from differences in duration rather than pure strength of activation.

Choice reaction times for temporal numerosity.

Subjects watched rapidly presented visual sequences of two events until they decided which event was the more frequent. Accuracy was reduced both by increased rates of presentation and by the addition of more inconclusive information to the initial portion of the sequence. Response latencies were longest when there was no objective numerosity inbalance, and they decreased, for both correct responses and errors, when discrimination was relatively easy. The patterns of decision latencies for individual subjects could be represented on a continuum defined by the sensitivity of the latencies to the rate of event presentation and to the objective inbalance between the two events. The discussion points out formal and empirical similarities between sequential numerosity judgments and choice reaction times. The present results are discussed within the framework of a first-passage time model, to be developed formally in a subsequent paper.

Cerebral dominance and asynchrony between bimanual two-dimensional movements.

The asynchrony of bimanual movements was investigated. Right- and left-handers traced simple geometrical patterns (ellipses) continuously with both hands. All combinations of the direction of rotation in each hand were executed at different rhythms. Geometrically, performances were largely independent of manual dominance. However, by comparing the passage times at homologous positions, the authors found that the dominant hand led the nondominant one by about 25 ms. The asynchrony was affected by neither movement type nor rhythm. The variability of the asynchrony varied along the trajectory, with well-defined maxima and minima. The variability profiles for movements that engaged homologous muscles differed markedly from those that engaged nonhomologous muscles. The authors discuss the hypothesis that bimanual periodic movements are timed by a lateralized functional module and asynchrony is due to the necessity of transmitting time-keeping information to the other hemisphere.

Biological movements look uniform: evidence of motor-perceptual interactions.

Six experiments demonstrate a visual dynamic illusion. Previous work has shown that in 2-dimensional (2D) drawing movements, tangential velocity and radius of curvature covary in a constrained manner. The velocity of point stimuli is perceived as uniform if and only if this biological constraint is satisfied. The illusion is conspicuous: The variations of velocity in the stimuli exceed 200%. Yet movements are perceived as uniform. Conversely, 2D stimuli moving at constant velocity are perceived as strongly nonuniform. The illusion is robust: Exposure to true constant velocity fails to suppress it. Results cannot be explained entirely by the kinetic depth effect. The illusion is evidence of a coupling between motor and perceptual processes: Even in the absence of any intention to perform a movement, certain properties of the motor system implicitly influence perceptual interpretation of the visual stimulus.

A developmental study of the relationship between geometry and kinematics in drawing movements.

Trajectory and kinematics of drawing movements are mutually constrained by functional relationships that reduce the degrees of freedom of the hand-arm system. Previous investigations of these relationships are extended here by considering their development in children between 5 and 12 years of age. Performances in a simple motor task--the continuous tracing of elliptic trajectories--demonstrate that both the phenomenon of isochrony (increase of the average movement velocity with the linear extent of the trajectory) and the so-called two-thirds power law (relation between tangential velocity and curvature) are qualitatively present already at the age of 5. The quantitative aspects of these regularities evolve with age, however, and steady-state adult performance is not attained even by the oldest children. The power-law formalism developed in previous reports is generalized to encompass these developmental aspects of the control of movement.

Minimum-jerk, two-thirds power law, and isochrony: converging approaches to movement planning.

Two approaches to the study of movement planning were contrasted. Data on the drawing of complex two-dimensional trajectories were used to test whether the covariations of the kinematic and geometrical parameters of the movement formalized by the two-thirds power law and by the isochrony principle (P. Viviani & R. Schneider, 1991) can be derived from the minimum-jerk model hypothesis (T. Flash & N. Hogan, 1985). The convergence of the 2 approaches was satisfactory insofar as the relation between tangential velocity and curvature is concerned (two-thirds power law). Global isochrony could not be deduced from the optimal control hypothesis. Scaling of velocity within movement subunits can instead be derived from the minimum-jerk hypothesis. The implications vis-à-vis the issue of movement planning are discussed with an emphasis on the representation used by the motor control system for coding the intended trajectories.

Segmentation and coupling in complex movements.

In two experiments we explore the structure of complex sequences of drawing movements. We find that in these movements a single parameter--the velocity gain factor--relates the geometrical and kinematic aspects of the movement trajectory via a two-thirds power law. In Experiment 1 we investigate the relation between the velocity gain factor and the linear extent of the trajectory. In Experiment 2 we demonstrate that the gain factor provides a criterion for segmenting the movement into distinct units of motor action, and we investigate the effects of the speed of execution on this segmentation. A theoretical analysis shows that the results of both Experiments 1 and 2 can be given a unitary interpretation by assuming a coupling function of variable strength between segments. The general problem of representing motor programs is discussed within this theoretical framework.

Frames of reference and control parameters in visuomanual pointing.

Three hypotheses concerning the control variables in visuomanual pointing were tested. Participants pointed to a visual target presented briefly in total darkness on the horizontal plane. The starting position of the hand alternated randomly among 4 points arranged as a diamond. Results show that during the experiment, movement drifted from hypometric to hypermetric. Final positions depended on the starting position. Their average pattern reproduced the diamond of the starting points, either in same orientation (hypometric trials), or with a double inversion (hypermetric trials). The distribution of variable errors was elliptical, with the major axis aligned with the direction of the movement. Statistical analysis and Monte Carlo simulations showed that the results are incompatible with the final point control hypothesis (A. Polit & E. Bizzi, 1979). Better, but not fully satisfactory, agreement was found with the view that pointing involves comparing initial and desired postures (J. F. Soechting & M. Flanders, 1989a). The hypothesis that accounted best for the results is that final hand position is coded as a vector represented in an extrinsic frame of reference centered on the hand.