The Effect of Eccentric or Isometric Training on Strength, Architecture, and Sprinting across an Australian Football Season.

PURPOSE: This study aimed to investigate the effect of an isometric (ISO) or Nordic hamstring exercise (NHE) intervention, alongside a sprint training program on hamstring strength, architecture, and sprinting performance in Australian footballers. METHODS: Twenty-five male athletes undertook NHE ( n = 13) or ISO ( n = 12) training across a 38-wk period (including preseason and in season). Biceps femoris long head (BFlh) architecture, ISO, and eccentric knee flexor strength were assessed at baseline, at the end of preseason (14 wk), and at the conclusion of the intervention. Sprint times and force-velocity profiles were determined at baseline and at the end of preseason. RESULTS: After the intervention, both groups had significant improvements in BFlh fascicle length (NHE: 1.16 cm, 95% CI = 0.68 to 1.63 cm, d = 1.88, P < 0.001; ISO: 0.82 cm, 95% CI = 0.57 to 1.06 cm, d = 1.70, P < 0.001), muscle thickness (NHE: 0.11 cm, 95% CI = 0.01 to 0.21 cm, d = 0.51, P = 0.032; ISO: 0.21 cm, 95% CI = 0.10 to 0.32 cm, d = 0.86, P = 0.002), and eccentric strength (NHE: 83 N, 95% CI = 53 to 114 N, d = 1.79, P < 0.001; ISO: 83 N, 95% CI = 17 to 151 N, d = 1.17, P = 0.018). Both groups also finished the intervention weaker isometrically than they started (NHE: -45 N, 95% CI = -81 to -8 N, d = -1.03, P = 0.022; ISO: -80 N, 95% CI = -104 to -56 N, d = -3.35, P < 0.001). At the end of preseason, the NHE group had improved their 5-m sprint time by 3.3% ± 2.0%), and their maximum horizontal velocity was 3% ± 2.1% greater than the ISO group who saw no changes. CONCLUSIONS: Both ISO and NHE training with a periodized sprinting program can increase BFlh fascicle length, thickness, and eccentric strength in Australian footballers. NHE training also improves 5-m sprint time and maximum velocity. However, both interventions reduced ISO strength. These findings provide unique, contextually relevant insights into the adaptations possible in semiprofessional athletes.

Utilization of mechanistic modelling and simulation to analyse fenspiride proarrhythmic potency - Role of physiological and other non-drug related parameters.

WHAT IS KNOWN AND OBJECTIVE: Fenspiride, a drug that had been used for decades for the treatment of respiratory diseases, was recently withdrawn from the market due to the potential risk of QT prolongation and proarrhythmia. This is the first such withdrawal for many years and hence poses a question whether such risk could have been predicted and to what degree non-drug-specific parameters play a role in the reported QT prolongation and cases of TdP. The study aim was to test various 'what-if' scenarios to assess the influence of age, gender, heart rate, and plasma potassium concentration on QT interval prolongation due to various doses of fenspiride with the use of mechanistic mathematical modelling. METHODS: Concentration-time profiles were simulated with the use of a PBPK model developed based on published physico-chemical data, data from in vitro ADME experiments, and in vivo PK study results. Pharmacodynamic effect, that is, drug-triggered pseudoECG signal modification was simulated using a biophysically detailed model of human cardiac myocytes. Analysis of the qNet metric was also performed to classify proarrhythmic risk related to fenspiride. RESULTS: In the simulation study, arrhythmia was not observed even in the 'what-if' scenarios with extreme exposure, age, heart rate, and plasma potassium concentration. The qNet metric value positioned fenspiride in the intermediate risk class. WHAT IS NEW AND CONCLUSION: It can be hypothesized that the clinically observed arrhythmia cases were not directly caused by fenspiride alone but a combination of multiple factors, including comedications.

Substantia nigra micro-haemorrhage causing ipsilateral unilateral Parkinsonism and abnormal dopamine transporter scan uptake.

Parkinsonism is a commonly seen movement disorder syndrome with neurodegenerative and non-neurodegenerative causes. Presynaptic dopamine transporter (DaT) single-photon emission computed tomography (SPECT) imaging is the most commonly used imaging technique in clinical practice to differentiate degenerative Parkinson's disease (PD) and PD plus syndromes from other causes such as essential tremor and drug-induced parkinsonism. This can help identify the patients who would benefit from medical therapy due to underlying pre-synaptic dopaminergic deficits. We report a case of unilateral parkinsonism caused by ipsilateral substantia nigra micro-haemorrhage resulting in disruption of the nigrostriatal pathway. This is an unusual case of a 55-year-old male patient who presented with unilateral Parkinsonism a decade after significant head trauma where MRI plays a critical and complementary role in diagnosing complete interruption of the nigrostriatal pathway due to cerebral micro-haemorrhage. The case also beautifully demonstrates the anatomy of the nigrostriatal pathway where a small lesion in the substantia nigra caused complete loss of radioligand uptake in the ipsilateral corpus striatum. Physicians should be aware of the importance of structural imaging in atypical movement disorder cases and, in particular, the routine use of susceptibility-weighted sequences (SWI).

Razor hamstring curl and Nordic hamstring exercise architectural adaptations: Impact of exercise selection and intensity.

OBJECTIVES: To investigate knee flexor strength and biceps femoris long head (BFlh) architectural adaptations following two different Nordic hamstring exercise (NHE) interventions and one razor hamstring curl (RHC) intervention. METHODS: Thirty recreationally active males performed a total of 128 reps of NHEbodyweight (n = 10), NHEweighted (n = 10), or RHCweighted training (n = 10) across 6 weeks. Following the intervention, participants avoided any eccentric training for 4 weeks (detraining period). Strength results during the NHE and RHC were recorded pre- and post-intervention, as well as following detraining. Architectural characteristics of the BFlh were assessed weekly throughout the intervention and detraining periods. RESULTS: For the NHEweighted group, NHE strength increased (+81N, P = 0.044, d = 0.90) and BFlh fascicles lengthened (+1.57 cm, P < 0.001, d = 1.41) after 6 weeks of training. After 1 week of detraining, BFlh fascicle lengths shortened, with the largest reductions seen in the NHEweighted group (-0.96 cm, P = 0.021, d = -0.90). Comparatively, BFlh fascicle length and NHE strength responses were moderate in the NHEbodyweight group and negligible in the RHCweighted group. The greatest RHC strength changes (+82N, P = 0.038, d = 1.15) were seen in the RHCweighted group. CONCLUSIONS: NHEweighted interventions induce large BFlh fascicle lengthening responses and these adaptations decay after just 1 week of detraining. NHEbodyweight training has a moderate impact on BFlh architecture while the RHCweighted group has the least. Weighted NHE and RHC training promoted exercise-specific increases in strength. These findings suggest that exercise selection and intensity should be considered when prescribing exercises aiming to increase eccentric strength and BFlh fascicle length.

Balint syndrome (chronic visual-spatial disorder) presenting without known cause.

Balint's syndrome is a rare disorder characterized by a triad of simultanagnosia, optic apraxia, and ocular apraxia. The syndrome manifests when there is an injury to the posterior parietal and occipital lobes and is often bilateral. Several causes of this syndrome were published in the literature, such as trauma, infarctions, infections, tumors, and pre-eclampsia. It can also be the presenting feature of several neurodegenerative disorders, such as atypical Alzheimer's disease. We report a case of a 62-year-old lady who presented with simultanagnosia, optic apraxia, and ocular ataxia which are the typical signs and symptoms of this syndrome. Neuropsychological evaluation revealed severe affection of the visual-spatial function with intact memory, language, and cognition. Brain imaging confirmed atrophy and decreased perfusion in the posterior parietal and occipital lobes. No underlying cause could be identified to explain the brain parenchymal atrophy. The follow-up neuropsychological assessment and brain imaging did not show any progression confirming the static course of the disease.

Running exposure is associated with the risk of hamstring strain injury in elite Australian footballers.

BACKGROUND: To investigate the association between running exposure and the risk of hamstring strain injury (HSI) in elite Australian footballers. METHODS: Elite Australian footballers (n=220) from 5 different teams participated. Global positioning system (GPS) data were provided for every athlete for each training session and match for the entire 2015 season. The occurrences of HSIs throughout the study period were reported. Receiver operator characteristic curve analyses were performed and the relative risk (RR) of subsequent HSI was calculated for absolute and relative running exposure variables related to distance covered above 10 and 24 km/hour in the preceding week/s. RESULTS: 30 prospective HSIs occurred. For the absolute running exposure variables, weekly distance covered above 24 km/hour (>653 m, RR=3.4, 95% CI 1.6 to 7.2, sensitivity=0.52, specificity=0.76, area under the curve (AUC)=0.63) had the largest influence on the risk of HSI in the following week. For the relative running exposure variables, distance covered above 24 km/hour as a percentage of distance covered above 10 km/hour (>2.5%, RR=6.3, 95% CI 1.5 to 26.7, sensitivity=0.93, specificity=0.34, AUC=0.63) had the largest influence on the risk of HSI in the following week. Despite significant increases in the RR of HSI, the predictive capacity of these variables was limited. CONCLUSIONS: An association exists between absolute and relative running exposure variables and elite Australian footballers' risk of subsequent HSI, with the association strongest when examining data within 7-14 days. Despite this, the use of running exposure variables displayed limited clinical utility to predict HSI at the individual level.

Meningoencephalitis in a Royal Marine after skinning reindeer in Norway.

Meningoencephalitis presenting in service personnel overseas may present a diagnostic challenge due to the broad range of potential differential diagnosis as well as the requirement for rapid assessment and treatment. A 25-year-old Royal Marine was evacuated to the Royal Centre for Defence Medicine in Birmingham, UK, with a history of rash consistent with erythema chronicum migrans, a seizure, and lymphocytic pleocytosis after skinning reindeer in Norway. Neuroborreliosis was suspected and empirical antibiotics were administered. Despite subsequent negative serology for Borrelia burgdorferi, given the clinical features and lymphocytic pleocytosis, an atypical presentation of neuroborreliosis remains a possible diagnosis in this scenario. This case serves to illustrate that British military personnel on exercise are potentially at risk of contracting borreliosis both in the UK and abroad, serological tests can be unreliable, and the differential diagnosis of meningoencephalitis can be broad with specialist input often required.

Predicting Drug Safety and Communicating Risk: Benefits of a Bayesian Approach.

Drug toxicity is a major source of attrition in drug discovery and development. Pharmaceutical companies routinely use preclinical data to predict clinical outcomes and continue to invest in new assays to improve predictions. However, there are many open questions about how to make the best use of available data, combine diverse data, quantify risk, and communicate risk and uncertainty to enable good decisions. The costs of suboptimal decisions are clear: resources are wasted and patients may be put at risk. We argue that Bayesian methods provide answers to all of these problems and use hERG-mediated QT prolongation as a case study. Benefits of Bayesian machine learning models include intuitive probabilistic statements of risk that incorporate all sources of uncertainty, the option to include diverse data and external information, and visualizations that have a clear link between the output from a statistical model and what this means for risk. Furthermore, Bayesian methods are easy to use with modern software, making their adoption for safety screening straightforward. We include R and Python code to encourage the adoption of these methods.

An Analysis of the Relationship Between Preclinical and Clinical QT Interval-Related Data.

There has been significant focus on drug-induced QT interval prolongation caused by block of the human ether-a-go-go-related gene (hERG)-encoded potassium channel. Regulatory guidance has been implemented to assess QT interval prolongation risk: preclinical guidance requires a candidate drug's potency as a hERG channel blocker to be defined and also its effect on QT interval in a non-rodent species; clinical guidance requires a "Thorough QT Study" during development, although some QT prolonging compounds are identified earlier via a Phase I study. Clinical, heart rate-corrected QT interval (QTc) data on 24 compounds (13 positives; 11 negatives) were compared with their effect on dog QTc and the concentration of compound causing 50% inhibition (IC50) of hERG current. Concordance was assessed by calculating sensitivity and specificity across a range of decision thresholds, thus yielding receiver operating characteristic curves of sensitivity versus (1-specificity). The area under the curve of ROC curves (for which 0.5 and 1 indicate chance and perfect concordance, respectively) was used to summarize concordance. Three aspects of preclinical data were compared with the clinical outcome (receiver operating characteristic area under the curve values shown in brackets): absolute hERG IC50 (0.78); safety margin between hERG IC50 and clinical peak free plasma exposure (0.80); safety margin between QTc effects in dogs and clinical peak free plasma exposure (0.81). Positive and negative predictive values of absolute hERG IC50 indicated that from an early drug discovery perspective, low potency compounds can be progressed on the basis of a low risk of causing a QTc increase.

From the Cover: High-Throughput Imaging of Cardiac Microtissues for the Assessment of Cardiac Contraction during Drug Discovery.

Cardiotoxicity is a common cause of attrition in preclinical and clinical drug development. Current in vitro approaches have two main limitations, they either are limited to low throughput methods not amendable to drug discovery or lack the physiological responses to allow an integrated risk assessment. A human 3D cardiac microtissue containing human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs), cardiac endothelial cells and cardiac fibroblast were used to assess their suitability to detect drug induced changes in cardiomyocyte contraction. These cardiac microtissues, have a uniform size, spontaneously beat, lack a hypoxic core, and contain key markers of each cell type. Application of field stimulation and measurement of cardiac contraction confirm cardiac microtissues to be a suitable model to investigate drug-induced changes in cardiomyocyte contractility. Using a bespoke image acquisition work flow and optical flow analysis method to test 29 inotroptic and 13 non-inotroptic compounds in vivo We report that cardiac microtissues provide a high-throughput experimental model that is both able to detect changes in cardiac contraction with a sensitivity and specificity of 80 and 91%, respectively, and provide insight into the direction of the inotropic response. Allowing improved in vitro cardiac contractility risk assessment. Moreover, our data provide evidence of the detection of this liability at therapeutically relevant concentrations with a throughput amenable to drug discovery.

Assessment of extracellular field potential and Ca2+ transient signals for early QT/pro-arrhythmia detection using human induced pluripotent stem cell-derived cardiomyocytes.

Cardiovascular toxicity is a prominent reason for failures in drug development, resulting in the demand for assays that can predict this liability in early drug discovery. We investigated whether iCell® cardiomyocytes have utility as an early QT/TdP screen. Thirty clinical drugs with known QT/TdP outcomes were evaluated blind using label-free microelectrode array (parameters measured were beating period (BP), field potential duration (FPD), fast Na+ amplitude and slope) and live cell, fast kinetic fluorescent Ca2+ transient FLIPR® Tetra (parameters measured were peak count, width, amplitude) systems. Many FPD-altering drugs also altered BP. Correction for BP, using a Log-Log (LL) model, was required to appropriately interpret direct drug effects on FPD. In comparison with human QT effects and when drug activity was to be predicted at top test concentration (TTC), LL-corrected FPD and peak count had poor assay sensitivity and specificity values: 13%/64% and 65%/11%, respectively. If effective free therapeutic plasma concentration (EFTPC) was used instead of TTC, the values were 0%/100% and 6%/100%, respectively. When compared to LL-corrected FPD and peak count, predictive values of uncorrected FPD, BP, width and amplitude were not much different. If pro-arrhythmic risk was to be predicted using Ca2+ transient data, the values were 67%/100% and 78%/53% at EFTPC and TTC, respectively. Thus, iCell® cardiomyocytes have limited value as an integrated QT/TdP assay, highlighting the urgent need for improved experimental alternatives that may offer an accurate integrated cardiomyocyte safety model for supporting the development of new drugs without QT/TdP effects.

Enhanced characterization of contractility in cardiomyocytes during early drug safety assessment.

We sought to investigate whether drug-induced changes in contractility were affected by pacing rates that represent the range of heart rates encountered in vivo. Using the cell geometry measurement system (IonOptix), we paced dog cardiomyocytes at different cycle lengths (CLs) of 2000, 1000, 500, and 333.3 ms, before and after exposure to 13 inotropic drugs. Time course data using vehicle control (0.1% dimethyl sulfoxide (DMSO)) demonstrated stability of the system at all CLs tested. Seven positive inotropes (eg isoproterenol) exerted rate-dependent increases in sarcomere shortening (Sarc. short.; maximal effect at a CL of 333.3 ms [0.1 µM isoproterenol increased Sarc. short. by 41.1% and 145.9% at 2000 and 333.3 ms, respectively]). Omecamtiv mecarbil showed an atypical profile (increased Sarc. short. at 2000 ms [106.9%] and decreased at 333.3 ms [IC(50) = 0.64 µM]). Four negative inotropes (eg flecainide) showed rate-independent inhibition of Sarc. short. (IC(50)s: 3.3 µM [2000 ms] versus 2.3 µM [333.3 ms]). The remaining negative inotropes, verapamil, and BTS (N-benzyl-p-toluene sulphonamide) produced an increase (IC(50)s: 3.9 µM [2000 ms] versus 0.043 µM [333.3ms]) and decrease (IC(50)s: 18.3 µM [2000 ms] versus 34.0 µM [333.3 ms]) in potency, respectively. Negative inotropes (eg flecainide, BTS, and verapamil) decreased the area of the Ca(2+) transient versus Sarc. short. hysteresis loop, although rate dependency was seen with verapamil only. Positive inotropes (eg isoproterenol and levosimendan) induced a rate-dependent increase in the area, however Omecamtiv mecarbil increased and decreased the area at CLs of 2000 and 333.3 ms, respectively. Thus, the use of different pacing rates may improve the detection of inotropes in early drug discovery and illustrate the potential for finger-printing different mechanisms of action.

Assessment of cardiomyocyte contraction in human-induced pluripotent stem cell-derived cardiomyocytes.

Functional changes to cardiomyocytes are a common cause of attrition in preclinical and clinical drug development. Current approaches to assess cardiomyocyte contractility in vitro are limited to low-throughput methods not amenable to early drug discovery. Human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) were used to assess their suitability to detect drug-induced changes in cardiomyocyte contraction. Application of field stimulation and measurement of cardiac contraction (IonOptix edge detection) and Ca(2+) transients confirmed hiPS-CMs to be a suitable model to investigate drug-induced changes in cardiomyocyte contractility. Using a live cell, fast kinetic fluorescent assay with a Ca(2+) sensitive dye to test 31 inotropic and 20 non-inotropic compounds in vivo, we report that hiPS-CMs provide a high-throughput experimental model to detect changes in cardiomyocyte contraction that is applicable to early drug discovery with a sensitivity and specificity of 87% and 70%, respectively. Moreover, our data provide evidence of the detection of this liability at therapeutically relevant concentrations with throughput amenable to influencing chemical design in drug discovery. Measurement of multiple parameters of the Ca(2+) transient in addition to the number of Ca(2+) transients offered no insight into the mechanism of cardiomyocyte contraction.

Reducing attrition in drug development: smart loading preclinical safety assessment.

Entry into the crucial preclinical good laboratory practice (GLP) stage of toxicology testing triggers significant R&D investment yet >20% of AstraZeneca's potential new medicines have been stopped for safety reasons in this GLP phase alone. How could we avoid at least some of these costly failures? An analysis of historical toxicities that caused stopping ('stopping toxicities') showed that >50% were attributable to target organ toxicities emerging within 2 weeks of repeat dosing or to acute cardiovascular risks. By frontloading 2-week repeat-dose toxicity studies and a comprehensive assessment of cardiovascular safety, we anticipate a potential 50% reduction in attrition in the GLP phase. This will reduce animal use overall, save significant R&D costs and improve drug pipeline quality.

Preservation of cardiomyocytes from the adult heart.

Cardiomyocytes represent one of the most useful models to conduct cardiac research. A single adult heart yields millions of cardiomyocytes, but these cells do not survive for long after isolation. We aimed to determine whether inhibition of myosin II ATPase that is essential for muscle contraction may preserve fully differentiated adult cardiomyocytes. Using inhibitors of the myosin II ATPase, blebbistatin and N-benzyl-p-toluene sulphonamide (BTS), we preserved freshly isolated fully differentiated adult primary cardiomyocytes that were stored at a refrigerated temperature. Specifically, preserved cardiomyocytes stayed viable for a 2-week period with a stable expression of cardiac genes and retained the expression of key markers characteristic of cardiomyocytes. Furthermore, voltage-clamp, action potential, calcium transient and contractility studies confirmed that the preserved cardiomyocytes are comparable to freshly isolated cells. Long-term exposure of preserved cardiomyocytes to four tyrosine kinase inhibitors, sunitinib malate, dasatinib, sorafenib tosylate and imatinib mesylate, revealed their potential to induce cardiac toxicity that was manifested with a decrease in contractility and induction of cell death, but this toxicity was not observed in acute experiments conducted over the time course amenable to freshly prepared cardiomyocytes. This study introduces the concept that the inhibition of myosin II ATPase safeguards the structure and function of fully differentiated adult cardiomyocytes. The fact that these preserved cardiomyocytes can be used for numerous days after preparation makes them a robust and versatile tool in cardiac research and allows the investigation of long-term exposure to novel drugs on cardiomyocyte function.

The response to rapid infusion of fentanyl in the human brain measured using pulsed arterial spin labelling.

OBJECTIVE: We evaluated the sensitivity of pulsed Arterial Spin Labelling (pASL) for the detection of changes in regional cerebral blood perfusion (CBP) during and after intra-venous (i.v.) infusion of an opioid agonist (fentanyl) and an opioid antagonist (naloxone). MATERIALS AND METHODS: Twenty-three subjects were scanned four times, receiving i.v. infusion of fentanyl, naloxone, placebo and a second fentanyl administration, in four separate scanning sessions in randomised order. End-tidal CO(2), respiration rate and heart rate were recorded continuously throughout each scan. pASL time series were collected using single shot EPI for 15 min (including 5 min of baseline prior to infusion). RESULTS: Significant increases in CBP were detected during and after administration of fentanyl, (when compared to placebo and naloxone), in most areas of high concentration of mu-opioid receptors (thalamus, lingual gyrus, para-hippocampal gyrus, and insula); near-significant increases were also observed in the insula. No increases in perfusion were observed during or after naloxone infusion. No correlation was found between regional rCBF changes and end-tidal CO(2), respiration rate or heart rate. Good reliability was found between the first and second fentanyl sessions but the regions of high reliability did not overlap completely with those of highest perfusion change. CONCLUSION: pASL is a suitable method for examining rapid, dynamic effects of opioid administration on brain physiology.