Minimal mechanical load and tissue culture conditions preserve native cell phenotype and morphology in tendon—a novel ex vivo mouse explant model

Appropriate mechanical load is essential for tendon homeostasis and optimal tissue function. Due to technical challenges in achieving physiological mechanical loads in experimental tendon model systems, the research community still lacks well‐characterized models of tissue homeostasis and physiological relevance. Toward this urgent goal, we present and characterize a novel ex vivo murine tail tendon explant model. Mouse tail tendon fascicles were extracted and cultured for 6 days in a load‐deprived environment or in a custom‐designed bioreactor applying low magnitude mechanical load (intermittent cycles to 1% strain, at 1 Hz) in serum‐free tissue culture. Cells remained viable, as did collagen structure and mechanical properties in all tested conditions. Cell morphology in mechanically loaded tendon explants approximated native tendon, whereas load‐deprived tendons lost their native cell morphology. These losses were reflected in altered gene expression, with mechanical loading tending to maintain tendon specific and matrix remodeling genes phenotypic of native tissue. We conclude from this study that ex vivo load deprivation of murine tendon in minimal culture medium results in a degenerative‐like phenotype. We further conclude that onset of tissue degeneration can be suppressed by low‐magnitude mechanical loading. Thus a minimal explant culture model featuring serum‐free medium with low mechanical loads seems to provide a useful foundation for further investigations. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1383–1390, 2018.

     

Evaluation of the Novel PET Tracer [11C]HACH242 for Imaging the GluN2B NMDA Receptor in Non-Human Primates

Purpose

There are currently no positron emission tomography (PET) radiotracers for the GluN2B (NR2B) binding sites of brain N-methyl-d-aspartate (NMDA) receptors. In rats, the GluN2B antagonist Ro25-6981 reduced the binding of N-((5-(4-fluoro-2-[¹¹C]methoxyphenyl)pyridin-3-yl)methyl)cyclopentanamin ([¹¹C]HACH242). This paper reports the evaluation of [¹¹C]HACH242 PET in non-human primates at baseline and following administration of the GluN2B negative allosteric modulator radiprodil.

Procedures

Eight 90-min dynamic [¹¹C]HACH242 PET scans were acquired in three male anaesthetised rhesus monkeys, including a retest session of subject 1, at baseline and 10 min after intravenous 10 mg/kg radiprodil. Standardised uptake values (SUV) were calculated for 9 brain regions. Arterial blood samples were taken at six timepoints to characterise pharmacokinetics in blood and plasma. Reliable input functions for kinetic modelling could not be generated due to variability in the whole-blood radioactivity measurements.

Results

[¹¹C]HACH242 entered the brain and displayed fairly uniform uptake. The mean (±?standard deviation, SD) T_max was 17?±?7 min in baseline scans and 24?±?15 min in radiprodil scans. The rate of radioligand metabolism in plasma (primarily to polar metabolites) was high, with mean parent fractions of 26?±?10 % at 20 min and 8?±?5 % at 85 min. Radiprodil increased [¹¹C]HACH242 whole-brain SUV in the last PET frame by 25 %, 1 %, 3 and 17 % for subjects 1, 2, 3 and retest of subject 1, respectively. The mean brain to plasma ratio was 5.4?±?2.6, and increased by 39 to 110 % in the radiprodil condition, partly due to lower parent plasma radioactivity of ?11 to ?56 %.

Conclusions

The present results show that [¹¹C]HACH242 has a suitable kinetic profile in the brain and low accumulation of lipophilic radiometabolites. Radiprodil did not consistently change [¹¹C]HACH242 brain uptake. These findings may be explained by variations in cerebral blood flow, a low fraction of specifically bound tracer, or interactions with endogenous NMDA receptor ligands at the binding site. Further experiments of ligand interactions are necessary to facilitate the development of radiotracers for in vivo imaging of the ionotropic NMDA receptor.

     

The effect of motivational interviewing on physical activity level among older adults: a systematic review and meta-analysis

Abstract

Aim: To determine the reported effect of randomized controlled trials (RCTs) using Motivational Interviewing (MI) to advance physical activity among older adults.

Methods: We searched for RCTs in MEDLINE, EMBASE, CINAHL, AgeLine, PsycINFO and Cochrane Library from inception until March, 2019. Identified trials that used MI for improving physical activity in community-dwelling older adults (≥65?years).

Results: From 5616 citations identified from the search, we included three trials (four publications). There was no evidence of a significant difference between the effect of MI and usual care on physical activity in older adults (Standard Mean Difference (SMD) -0.02, 95% Confidence Interval (CI) 0.05 to 0.46, I² 16%; 3 trials; 84 participants).

Conclusion: There is insufficient evidence to support the effect of MI on improving physical activity among older adults. There is a need for more high quality trials to show that MI is beneficial in older adults who are physically inactive.     

Effect of Cyclosporine and Rifampin on the Pharmacokinetics of Macitentan, a Tissue-Targeting Dual Endothelin Receptor Antagonist

Macitentan is a dual endothelin receptor antagonist under phase 3 investigation in pulmonary arterial hypertension. We investigated the effect of cyclosporine (Cs) and rifampin on the pharmacokinetics of macitentan and its metabolites ACT-132577 and ACT-373898 in healthy male subjects. In addition, in vitro studies were performed to investigate interactions between macitentan and its active metabolite ACT-132577 with human organic anion-transporting polypeptides (OATPs). The clinical study (AC-055-111) was conducted as a two-part, one-sequence, crossover study. Ten subjects in each part received multiple-dose macitentan followed by multiple-dose co-administration of Cs (part A) or rifampin (part B). In the presence of Cs, steady-state area under the plasma concentration-time profiles during a dose interval (AUC(τ)) for macitentan and ACT-373898 increased 10% and 7%, respectively, and decreased 3% for ACT-132577. Steady-state AUC(τ) of macitentan and ACT-373898 in the presence of rifampin decreased 79% and 64%, respectively. For ACT-132577, no relevant difference in AUC(τ) between the two treatments was observed. Macitentan co-administered with Cs or rifampin was well tolerated. The complementary in vitro studies demonstrated no marked differences in uptake rates of macitentan and ACT-132577 between the wild-type and OATP over-expressing cells over the concentration range tested. Concomitant treatment with Cs did not have any clinically relevant effect on the exposure to macitentan or its metabolites, at steady-state. Concomitant treatment with rifampin reduced significantly the exposure to macitentan and its metabolite ACT-373898 at steady-state but did not affect the exposure to the active metabolite ACT-132577 to a clinically relevant extent.     

Integrated pharmacokinetics and pharmacodynamics of epoprostenol in healthy subjects

AIM

The aim of the study was to report the first thorough characterization of the pharmacokinetics (PK) and pharmacodynamics (PD) of epoprostenol in an integrated manner.

METHOD

Twenty healthy male subjects received two formulations of i.v. epoprostenol, in a crossover design, in sequential infusions of 2, 4, 6 and 8 ng kg⁻¹ min⁻¹ for 2 h each. A sensitive assay was developed which allowed accurate PK characterization of epoprostenol via analysis of the concentration–time profiles of its two primary metabolites, 6-keto-prostacyclin F_1α and 6,15-diketo-13,14-dihydro-prostacyclin F_1α. PD parameters included cardiac output (CO), cardiac index (CIn) and heart rate (HR).

RESULTS

The pharmacokinetics of epoprostenol deviated slightly from dose-proportionality, probably due to a food effect. After infusion of the two formulations of epoprostenol, the t_1/2 values expressed as geometric mean (95% confidence interval) were 0.25 h (0.14, 0.46) and 0.22 h (0.13, 0.38) for 6-keto-prostacyclin F_1α, and 0.32 h (0.22, 0.45) and 0.34 h (0.26, 0.46) for 6,15-diketo-13,14-dihydro-prostacyclin F_1α. A single compartment infusion model with first order elimination adequately described the PK of 6-keto-prostacyclin F_1α. This model also characterized the food effect. Stepwise infusions with epoprostenol resulted in a progressive increase in CO, CIn and HR.

CONCLUSION

Of the two metabolites analyzed, the appearance of 6-keto-prostacyclin F_1α in plasma was more closely associated with the haemodynamic effects of i.v. epoprostenol. PK and PD profiles showed that CIn relates proportionally and linearly to the plasma concentrations of 6-keto-prostacyclin F_1α. These results suggest that 6-keto-prostacyclin F_1α is a suitable surrogate marker of plasma concentrations of epoprostenol.     

Prior Mating Experience Modulates the Dispersal of <Emphasis Type="Italic">Drosophila</Emphasis> in Males More Than in Females

Cues from both an animal’s internal physiological state and its local environment may influence its decision to disperse. However, identifying and quantifying the causative factors underlying the initiation of dispersal is difficult in uncontrolled natural settings. In this study, we automatically monitored the movement of fruit flies and examined the influence of food availability, sex, and reproductive status on their dispersal between laboratory environments. In general, flies with mating experience behave as if they are hungrier than virgin flies, leaving at a greater rate when food is unavailable and staying longer when it is available. Males dispersed at a higher rate and were more active than females when food was unavailable, but tended to stay longer in environments containing food than did females. We found no significant relationship between weight and activity, suggesting the behavioral differences between males and females are caused by an intrinsic factor relating to the sex of a fly and not simply its body size. Finally, we observed a significant difference between the dispersal of the natural isolate used throughout this study and the widely-used laboratory strain, Canton-S, and show that the difference cannot be explained by allelic differences in the foraging gene.     

Evaluative conditioning of fear and disgust in blood-injection-injury phobia: specificity and impact of individual differences in disgust sensitivity

The present study examines whether the repeated pairing of neutral facial expressions with phobic-relevant stimuli differentially influences evaluative ratings of fear and disgust between analogue blood-injection-injury (BII) phobic (n=40) and non-phobic (n=40) participants. Consistent with prior research, BII phobics reported greater disgust sensitivity than non-phobic participants even after controlling for between group differences in anxiety symptoms. Results from the evaluative conditioning experiment indicated that pre- to posttest increases in fear ratings were only marginally greater for phobic compared to non-phobic participants. However, increases in disgust from pre- to posttest were greater for phobic compared to non-phobic participants and greater for neutral expressions that were paired with threat-relevant stimuli compared to stimuli not paired with threat-relevant stimuli. Subsequent analysis also indicated that pre- to posttest increases in disgust ratings of neutral expressions that were paired with threat-relevant stimuli was moderated by disgust sensitivity levels among phobic and non-phobic participants. Heightened fear and disgust ratings were subsequently reduced by an extinction procedure. Implications of present findings in understanding the role of fear and disgust in BII phobia are discussed.