Barriers to vaccine use in small ruminants and poultry in Tanzania.

Vaccination is an important disease prevention and control measure; however, vaccine adoption by livestock farmers in Tanzania is still low. This cross-sectional study examined the challenges to vaccine use faced by livestock owners and animal health professionals (AHPs) in Tanzania. A questionnaire was administered to 216 households that kept small ruminants and poultry and 19 AHPs' data were collected electronically via the survey platform Qualtrics, and descriptive statistics were performed. Households with poultry reported vaccinating mostly against Newcastle disease (91.7%), fowl pox (48.1%) and Gumboro disease (37.0%), whilst households with small ruminants reported contagious caprine pleuropneumonia (62.2%), sheep and goat pox (17.1%), foot-and-mouth disease (7.3%) and peste des petits ruminants (7.3%). The households' decision to vaccinate was mostly influenced by knowledge of diseases (82.4%), disease history on the farm (69.4%) and vaccine price (63.4%). Most households (54.6%) experienced challenges when purchasing vaccines, including high vaccine cost (78.0%), long distance from vaccine source (61.0%) and vaccine unavailability (21.2%). The findings suggest that improving the knowledge of livestock owners regarding the priority diseases and the benefits of vaccination, establishing more vaccine suppliers, improving vaccine distribution and access and training AHPs and households on appropriate vaccine storage and handling are necessary to improve vaccine adoption and ensure vaccine quality and effectiveness.

Prediction of Paroxysmal Atrial Fibrillation From Complexity Analysis of the Sinus Rhythm ECG: A Retrospective Case/Control Pilot Study.

Paroxysmal atrial fibrillation (PAF) is the most common cardiac arrhythmia, conveying a stroke risk comparable to persistent AF. It poses a significant diagnostic challenge given its intermittency and potential brevity, and absence of symptoms in most patients. This pilot study introduces a novel biomarker for early PAF detection, based upon analysis of sinus rhythm ECG waveform complexity. Sinus rhythm ECG recordings were made from 52 patients with (n = 28) or without (n = 24) a subsequent diagnosis of PAF. Subjects used a handheld ECG monitor to record 28-second periods, twice-daily for at least 3 weeks. Two independent ECG complexity indices were calculated using a Lempel-Ziv algorithm: R-wave interval variability (beat detection, BD) and complexity of the entire ECG waveform (threshold crossing, TC). TC, but not BD, complexity scores were significantly greater in PAF patients, but TC complexity alone did not identify satisfactorily individual PAF cases. However, a composite complexity score (h-score) based on within-patient BD and TC variability scores was devised. The h-score allowed correct identification of PAF patients with 85% sensitivity and 83% specificity. This powerful but simple approach to identify PAF sufferers from analysis of brief periods of sinus-rhythm ECGs using hand-held monitors should enable easy and low-cost screening for PAF with the potential to reduce stroke occurrence.

ECG Restitution Analysis and Machine Learning to Detect Paroxysmal Atrial Fibrillation: Insight from the Equine Athlete as a Model for Human Athletes.

Atrial fibrillation is the most frequent arrhythmia in both equine and human athletes. Currently, this condition is diagnosed via electrocardiogram (ECG) monitoring which lacks sensitivity in about half of cases when it presents in paroxysmal form. We investigated whether the arrhythmogenic substrate present between the episodes of paroxysmal atrial fibrillation (PAF) can be detected using restitution analysis of normal sinus-rhythm ECGs. In this work, ECG recordings were obtained during routine clinical work from control and horses with PAF. The extracted QT, TQ, and RR intervals were used for ECG restitution analysis. The restitution data were trained and tested using k-nearest neighbor (k-NN) algorithm with various values of neighbors k to derive a discrimination tool. A combination of QT, RR, and TQ intervals was used to analyze the relationship between these intervals and their effects on PAF. A simple majority vote on individual record (one beat) classifications was used to determine the final classification. The k-NN classifiers using two-interval measures were able to predict the diagnosis of PAF with area under the receiving operating characteristic curve close to 0.8 (RR, TQ with k ≥ 9) and 0.9 (RR, QT with k ≥ 21 or TQ, QT with k ≥ 25). By simultaneously using all three intervals for each beat and a majority vote, mean area under the curves of 0.9 were obtained for all tested k-values (3-41). We concluded that 3D ECG restitution analysis can potentially be used as a metric of an automated method for screening of PAF.

The complexity of clinically-normal sinus-rhythm ECGs is decreased in equine athletes with a diagnosis of paroxysmal atrial fibrillation.

Equine athletes have a pattern of exercise which is analogous to human athletes and the cardiovascular risks in both species are similar. Both species have a propensity for atrial fibrillation (AF), which is challenging to detect by ECG analysis when in paroxysmal form. We hypothesised that the proarrhythmic background present between fibrillation episodes in paroxysmal AF (PAF) might be detectable by complexity analysis of apparently normal sinus-rhythm ECGs. In this retrospective study ECG recordings were obtained during routine clinical work from 82 healthy horses and from 10 horses with a diagnosis of PAF. Artefact-free 60-second strips of normal sinus-rhythm ECGs were converted to binary strings using threshold crossing, beat detection and a novel feature detection parsing algorithm. Complexity of the resulting binary strings was calculated using Lempel-Ziv ('76 & '78) and Titchener complexity estimators. Dependence of Lempel-Ziv '76 and Titchener T-complexity on the heart rate in ECG strips obtained at low heart rates (25-60 bpm) and processed by the feature detection method was found to be significantly different in control animals and those diagnosed with PAF. This allows identification of horses with PAF from sinus-rhythm ECGs with high accuracy.

The application of Lempel-Ziv and Titchener complexity analysis for equine telemetric electrocardiographic recordings.

The analysis of equine electrocardiographic (ECG) recordings is complicated by the absence of agreed abnormality classification criteria. We explore the applicability of several complexity analysis methods for characterization of non-linear aspects of electrocardiographic recordings. We here show that complexity estimates provided by Lempel-Ziv '76, Titchener's T-complexity and Lempel-Ziv '78 analysis of ECG recordings of healthy Thoroughbred horses are highly dependent on the duration of analysed ECG fragments and the heart rate. The results provide a methodological basis and a feasible reference point for the complexity analysis of equine telemetric ECG recordings that might be applied to automate detection of equine arrhythmias in equine clinical practice.

Characterization of recombinant hERG K(+) channel inhibition by the active metabolite of amiodarone desethyl-amiodarone.

UNLABELLED: The aim of this study was to determine the effects of desethyl-amiodarone (DEA), the major metabolite of the class III antiarrhythmic drug amiodarone, on human ether-à-go-go-related gene (hERG) encoded potassium channel current. MATERIALS AND METHODS: Whole-cell patch clamp recordings were made at 37 degrees C of ionic current (I(hERG)) carried by recombinant hERG channels expressed in HEK-293 cells. RESULTS: Desethyl-amiodarone inhibited I(hERG) with a half-maximal inhibitory concentration of approximately 158 nmol/L, compared with approximately 47 nmol/L for amiodarone. The inhibitory action of DEA on I(hERG) was contingent on channel gating, showing significant time and voltage dependence. Desethyl-amiodarone also produced an approximately -9 mV shift in the voltage dependence of activation of I(hERG); however, there was no significant preference for activated over inactivated channels. CONCLUSIONS: Because hERG underlies native cardiac "I(Kr)" channels, hERG/I(Kr) inhibition by DEA as well as amiodarone may contribute to the overall effects of amiodarone administration on cardiac repolarization.

Suppression of TRPC3 leads to disappearance of store-operated channels and formation of a new type of store-independent channels in A431 cells.

In most non-excitable cells, calcium (Ca(2+)) release from the inositol 1,4,5-trisphosphate (InsP(3))-sensitive intracellular Ca(2+) stores is coupled to Ca(2+) influx through the plasma membrane Ca(2+) channels whose molecular composition is poorly understood. Several members of mammalian TRP-related protein family have been implicated to both receptor- and store-operated Ca(2+) influx. Here we investigated the role of the native transient receptor potential 3 (TRPC3) homologue in mediating the store- and receptor-operated calcium entry in A431 cells. We show that suppression of TRPC3 protein levels by small interfering RNA (siRNA) leads to a significant reduction in store-operated calcium influx without affecting the receptor-operated calcium influx. With single-channel analysis, we further demonstrate that reduction of TRPC3 levels results in suppression of specific subtype of store-operated calcium channels and activation of store-independent channels. Our data suggest that TRPC3 is required for the formation of functional store-operated channels in A431 cells.

Functional properties of endogenous receptor- and store-operated calcium influx channels in HEK293 cells.

Activation of phospholipase C (PLC)-mediated signaling pathways in non-excitable cells causes the release of calcium (Ca2+) from inositol 1,4,5-trisphosphate (InsP3)-sensitive intracellular Ca2+ stores and activation of Ca2+ influx via plasma membrane Ca2+ channels. The properties and molecular identity of plasma membrane Ca2+ influx channels in non-excitable cells is a focus of intense investigation. In the previous studies we used patch clamp electrophysiology to describe the properties of Ca2+ influx channels in human carcinoma A431 cell lines. Now we extend our studies to human embryonic kidney HEK293 cells. By using a combination of Ca2+ imaging and whole cell and single channel patch clamp recordings we discovered that: 1) HEK293 cells contain four types of plasma membrane Ca2+ influx channels: I(CRAC), Imin, Imax, and I(NS); 2) I(CRAC) channels are highly Ca2+-selective (P(Ca/Cs)>1000) and I(CRAC) single channel conductance is too small for single channel analysis; 3) Imin channels in HEK293 cells display functional properties identical to Imin channels in A431 cells, with single channel conductance of 1.2 pS for divalent cations, 10 pS for monovalent cations, and divalent cation selectivity P(Ba/K)=20; 4) Imin channels in HEK293 cells are activated by InsP3 and inhibited by phosphatidylinositol 4,5-bisphosphate, but store-independent; 5) when compared with Imin, Imax channels have higher conductance for divalent (17 pS) and monovalent (33 pS) cations, but less selective for divalent cations (P(Ba/K)=4), 6) Imax channels in HEK293 cells can be activated by InsP3 or by Ca2+ store depletion; 7) I(NS) channels are non-selective (P(Ba/K)=0.4) and display a single channel conductance of 5 pS; and 8) I(NS) channels are not gated by InsP3 but activated by depletion of intracellular Ca2+ stores. Our findings provide novel information about endogenous Ca2+ channels supporting receptor-operated and store-operated Ca2+ influx pathways in HEK293 cells.