NcGRA2 as a molecular target to assess the parasiticidal activity of toltrazuril against Neospora caninum

The treatment of Neospora caninum infection in the bovine host is still at an experimental stage. In contrast to the in vivo situation, a wide range of compounds have been intensively investigated in cell-culture-based assays. Tools to demonstrate efficacy of treatment have remained conventional including morphological and cell biological criteria. In this work, we present a molecular assay that allows the distinction between live and dead parasites. Live parasites can be detected by measuring the mRNA level of specific genes, making use of the specific mRNA available in live cells. The NcGra2 gene of N. caninum, which is known to be expressed in both tachyzoites and bradyzoites, was used to establish a quantitative real-time RT-PCR, for monitoring parasite viability. Validation of the system in vitro was achieved using Neospora-infected cells that had been treated for 2-20 days with 30μg/ml toltrazuril. NcGRA2-RT-real time PCR demonstrated that a 10-day toltrazuril-treatment exerted parasitostatic activity, as assessed by the presence of NcGRA2-transcripts, whereas after a 14-day treatment period no NcGRA2-transcripts were detected, showing that the parasites were no longer viable. Concurrently, extended culture for a period of 4 weeks in the absence of the drug following the 14-day toltrazuril treatment did not lead to further parasite proliferation, confirming the parasiticidal effect of the treatment. This assay has the potential to be widely used in the development of novel drugs against N. caninum, with a view to distinguishing between parasiticidal and parasitostatic efficacy of given compound

Randomized trial comparing the i-gel™ and Magill tracheal tube with the single-use ILMA™ and ILMA™ tracheal tube for fibreoptic-guided intubation in anaesthetized patients with a predicted difficult airway

Background The i-gel™ is a single-use supraglottic airway device (SAD) that allows fibreoptic-guided tracheal intubation through the device. Until now, no prospective data for this procedure are available. Therefore, in a prospective randomized controlled trial, we evaluated fibreoptic-guided tracheal intubation with a standard Rüsch™ PVC tracheal tube (TT) through the i-gel™ compared with the single-use ILMA™ (sILMA™) TT through the sILMA™ in patients with a predicted difficult airway. Methods With ethics committee approval and written informed consent, 160 patients were randomly assigned to either SAD. After placement of the SAD, a fibreoptic bronchoscope was introduced into the trachea as a railroad for the TT. Primary outcome variable was the first-attempt fibreoptic-guided intubation success rate. Secondary variables included time for insertion and intubation, airway leak pressures, fibreoptic view, and adverse events. Data are presented as mean (sd) or percentages. A P-value of <0.05 was considered statistically significant. Results Fibreoptic-guided intubation was successful at the first attempt in 76 patients (96%) using the i-gel™ and in 71 patients (90%) using the sILMA™ (P=0.21). Most of the failed intubations were rescued by conventional laryngoscopy. Airway leak pressure was higher for the sILMA™. There were no problems during removal of either type of SAD. Conclusions Fibreopic-guided tracheal intubation through the i-gel™ using a standard Rüsch™ Magill TT is successful and an alternative to the sILMA™ with the sILMA™ T

Toltrazuril treatment of congenitally acquired Neospora caninum infection in newborn mice

C57BL/6 mice were infected with Neospora caninum tachyzoites during pregnancy, yielding a transplacental infection of developing fetuses. Subsequently, congenitally infected newborn mice were treated either once or three times with toltrazuril (or placebo) at a concentration of 31.25mg compound per kg body weight. Both toltrazuril and placebo treatment had no negative effect on newborns, as noninfected treated pups developed normally without differences in mortality and morbidity to matching nontreated control animals. Already one application of toltrazuril was significantly (p < 0.01) able to delay the outbreak of neosporosis in newborn mice, when compared to placebo-treated infected controls. We found significantly higher proportion of surviving newborns in one-time-toltrazuril-treated and three-time-toltrazuril-treated groups (34% and 54%, respectively) when compared to one-time-placebo-treated and three-time-placebo-treated groups (14% and 30%, respectively). There was no significant difference (p = 0.2) in the proportion of surviving pups between one-time-toltrazuril and three-time-toltrazuril treatment. However, the number of diseased and Neospora-positive pups (46% and 47%, respectively) was markedly reduced after three-time-toltrazuril treatment compared to all other groups. Three-time-treatment also resulted in the highest antibody (IgG, IgG2a) response. Pharmacokinetic analyses using individual serum samples revealed that, although toltrazuril was absorbed and metabolized to toltrazuril sulfone by newborn mice, medicated animals exhibited an unexpected rapid turn-over (half-life time) of the compound. Toltrazuril and the metabolite were also found in brain tissues, indicating that passage of the blood-brain barrier occurred. In conclusion, we could show that three times treatment with toltrazuril had a high impact on the course of infection in congenitally N. caninum-infected newborn mic

Recent Decisions

Comments on recent decisions by John M. Sullivan, William G. Greif, Joseph F. MacKrell, William N. Antonis, Thomas Meaney, Jr., William J. Hurley, Joseph H. Harrison, Robert L. Berry, Robert F. McCoy, Edward Canary, Maynard R. Bissonnette, and Luke R. Morin

i-gel™ supraglottic airway in clinical practice: a prospective observational multicentre study

Background The i-gel™ supraglottic airway device has been studied in randomized controlled studies, but it has not been evaluated in a large prospective patient cohort. Therefore, we performed this prospective multicentre observational study to evaluate success rates, airway leak pressure, risk factors for i-gel failure, and adverse events. Methods With Ethics Committee approval and waiver of patients' consent, data about anaesthesia providers, patient characteristics, and the performance of the i-gel were recorded in five independent hospitals in Switzerland over a period of 24 months. We analysed success rates, leak pressures, adverse events, and risk factors for failure. Results Data from 2049 i-gel uses were analysed. Patients' mean age was 47 (range 6-91) yr. The primary i-gel success rate without changing size was 93%; the overall success rate was 96%. Insertion was deemed very easy or easy in 92%. The mean airway leak pressure was 26 (8) cm H2O. The mean anaesthesia time was 67 (42) min. Risk factors associated with i-gel failure were males (P<0.001), impaired mandibular subluxation (P=0.01), poor dentition (P=0.02), and older age (P<0.01). Adverse events recorded were laryngeal spasms (n=25, 1.2%), blood stained airway devices (n=79, 3.9%), transient nerve damage (n=2, 0.1%), one case of transient vasovagal asystole, and one glottic haematoma. Conclusions The i-gel is a reliable supraglottic airway device failing in <5% and providing high airway leak pressures. Males, impaired mandibular subluxation, poor dentition, and older age are risk factors associated with primary device failure. Serious adverse events are rar

Randomized clinical trial of the i-gel™ and Magill tracheal tube or single-use ILMA™ and ILMA™ tracheal tube for blind intubation in anaesthetized patients with a predicted difficult airway

Background The single-use supraglottic airway device i-gel™ has been described in several case reports as a conduit for intubation, but no prospective data about success rates of blind intubation are available. Therefore, we performed this prospective randomized controlled trial to compare the success rate of blind tracheal intubation with a Magill PVC tube through the i-gel™ with intubation using an sILMA™ PVC tube through the single-use intubating laryngeal mask airway (sILMA™). Methods With ethics committee approval and written informed consent, 80 patients with predictors of a difficult airway were computer randomized to either supraglottic airway device (SAD). The corresponding tracheal tube (TT) was introduced through the SAD under fibreoptic visualization but without fibreoptic guidance. Primary outcome was blind intubation success rate. Times, airway leak pressure, fibreoptic view, and adverse events were recorded. To control for the influence of the TT, we compared data from 40 patients described in an accompanying study (sILMA™ with Magill TT and i-gel™ with sILMA™ TT). Results Blind intubation success rate through the sILMA™ (69%) was higher than with the i-gel™ (15%, P<0.001). Data from the other patient group excluded the TT type as the primary cause for the difference in success rate. Removal of SADs was without problems with no difference between the type of SAD. Conclusions Blind tracheal intubation using the sILMA™ tube through the sILMA™ is much more successful than blind intubation with a Magill PVC tube through the i-gel™. Because of its low success rate, we would not recommend blind intubation through the i-gel

Using Fidelity Measures to Support the Training of Graduate Students Working in School Settings

This paper presents a case example of a school-based social development program supported by counselors, yet implemented by graduate student facilitators in a mid-sized urban public school in New England. The authors describe a process for developing and validating fidelity instruments and the use of these assessments to train lay facilitators or graduate students enrolled in counseling or counselor education programs. Guidelines are presented for school counselors interested in using customized fidelity instruments to formatively assess training needs of graduate students

Quantitative Determination of Temperature in the Approach to Magnetic Order of Ultracold Fermions in an Optical Lattice

We perform a quantitative simulation of the repulsive Fermi-Hubbard model using an ultracold gas trapped in an optical lattice. The entropy of the system is determined by comparing accurate measurements of the equilibrium double occupancy with theoretical calculations over a wide range of parameters. We demonstrate the applicability of both high-temperature series and dynamical mean-field theory to obtain quantitative agreement with the experimental data. The reliability of the entropy determination is confirmed by a comprehensive analysis of all systematic errors. In the center of the Mott insulating cloud we obtain an entropy per atom as low as 0.77k(B) which is about twice as large as the entropy at the Neel transition. The corresponding temperature depends on the atom number and for small fillings reaches values on the order of the tunneling energy

Using Fidelity Measures to Support the Training of Graduate Students Working in School Settings

This paper presents a case example of a school-based social development program supported by counselors, yet implemented by graduate student facilitators in a mid-sized urban public school in New England. The authors describe a process for developing and validating fidelity instruments and the use of these assessments to train lay facilitators or graduate students enrolled in counseling or counselor education programs. Guidelines are presented for school counselors interested in using customized fidelity instruments to formatively assess training needs of graduate students