In Vitro Assessment of Cardiac Fibroblast Activation at Physiologic Stiffness.

Cardiac fibroblasts (CF) are an essential cell type in cardiac physiology, playing diverse roles in maintaining structural integrity, extracellular matrix (ECM) synthesis, and tissue repair. Under normal conditions, these cells reside in the interstitium in a quiescent state poised to sense and respond to injury by synthesizing and secreting collagen, vimentin, hyaluronan, and other ECM components. In response to mechanical and chemical stimuli, these "resident" fibroblasts can undergo a transformation through a continuum of activation states into what is commonly known as a "myofibroblast," in a process critical for injury response. Despite progress in understanding the contribution of fibroblasts to cardiac health and disease, much remains unknown about the signaling mediating this activation, in part owing to technical challenges in evaluating CF function and activation status in vitro. Given their role in monitoring the ECM, CFs are acutely sensitive to stiffness and pressure. High basal activation of isolated CFs is common due to the super-physiologic stiffness of traditional cell culture substrates, making assays dependent on quiescent cells challenging. To overcome this problem, cell culture parameters must be tightly controlled, and the use of dishes coated with biocompatible reduced-stiffness substrates, such as 8-kPa polydimethylsiloxane (PDMS), has shown promise in reducing basal activation of fibroblasts. Here, we describe cell culture protocol for maintaining CF quiescence in vitro to enable a dynamic range for the assessment of activation status in response to fibrogenic stimuli using PDMS-coated coverslips. Our protocol provides a cost-effective tool to study fibroblast signaling and activity, allowing researchers to better understand the underlying mechanisms involved in cardiac fibrosis. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Generation of 8-kPa polydimethylsiloxane (PDMS)/gelatin-coated coverslips for cardiac fibroblast cell culture Basic Protocol 2: Isolation of adult cardiac fibroblasts and plating onto PDMS coverslips Basic Protocol 3: Assessment of cardiac fibroblast activation by α smooth muscle actin (αSMA) immunocytochemistry.

Dopamine Concentration Changes Associated with the Retrodialysis of Methylone and 3,4-Methylenedioxypyrovalerone (MDPV) into the Caudate Putamen.

Structural modifications to synthetic psychoactive cathinones (SPCs), a class of drugs that contain a ß-keto modification of the phenethylamine pharmacophore of amphetamine, induce differences in dopamine transporter (DAT) activity. Here, in vivo retrodialysis was utilized to deliver the SPCs 3,4-methylenedioxypyrovalerone (MDPV, a DAT inhibitor) or methylone (a DAT substrate) into the caudate putamen of male Sprague-Dawley rats. Dialysate samples were collected prior to and post drug administration, and temporal changes in dopamine concentration were quantified using HPLC-EC methods. Methylone elicited a 200% increase and MDPV a 470% increase in dopamine levels at the 10 min time point. The findings demonstrate that in vivo retrodialysis can be used to evaluate the effects of SPCs on neurotransmission in the brain.

Altered reward sensitivity to sucrose outcomes prior to drug exposure in alcohol preferring rats.

Addiction involves key impairments in reward sensitivity (RS). The current study explored impaired RS to natural reward as a predisposing factor to addictive-like behavior. Alcohol preferring (P) rats are selectively bred based on significantly greater ethanol consumption and preference and offer the ability to inspect differences in subjects with a positive family history of addictive-like behavior. P rat's RS was compared to RS in the well-used Sprague-Dawley (SD) strain. To assess RS in a novel manner, instrumental incentive contrast, discrimination and consumption of sucrose solution were examined. Animals performed in a free operant situation for different sucrose concentration solutions using a block of 'mixed' trials with alternating outcome concentrations (e.g., 5 and 10 % sucrose) to change outcome value in a predictable manner. Animals also performed for reward in blocks of single outcome trials (5 or 10 or 20 or 40 % sucrose daily exposure) surrounding the mixed block. RS (e.g., reward discrimination and contrast effects between and within-sessions) was measured by changes in trials completed, instrumental response latency and consumption. P rats expressed an altered profile of RS with a greater tendency toward equivalent responding to different outcomes within the same session and an absence of incentive contrast from diverse reward comparisons. In contrast, SD animals expressed within-session reward discrimination and a subset of incentive contrast effects. These effects were moderated by food deprivation more consistently in SD compared to P rats. P rat alterations in processing natural rewards could predispose them to addictive-like behaviors including greater alcohol consumption and preference.

Reversal of temperature responses to methylone mediated through bi-directional fecal microbiota transplantation between hyperthermic tolerant and naïve rats.

The synthetic cathinone ("bath salt") methylone induces a hyperthermia response and with chronic administration tolerance to this hyperthermia has been reported. The microbiome-gut-brain axis has been implicated in multiple bodily systems and pathologies, and intentional manipulation of the gut-microbiome has yielded clinically significant results. Here, we examined the effects of bi-directional Fecal Microbiota Transplantation (FMT) between methylone-induced hyperthermic tolerant (MHT) and methylone-naïve (MN) rats. Rats treated with methylone once per week developed tolerance to methylone-induced hyperthermia by the fourth week. Once tolerant, daily bi-directional FMT between the two groups were performed for seven days prior to the next methylone treatment. The FMT abated the developed tolerance in the MHT group. When treated with methylone for the first time following FMT, recipient MN rats displayed significant tolerance to hyperthermia despite it being their initial drug treatment. Post-FMT, MHT rats displayed elevations in norepinephrine and expression of UCP1, UCP3 and TGR5 in brown adipose tissue, with reductions in expression of TGR5 and UCP3 in skeletal muscle. The pre- and post-FMT methylone tolerance phenotypes of transplant recipients are concurrent with changes in the relative abundance of several classes of Proteobacteria, most evident for Gammaproteobacteria and Alphaproteobacteria. MHT recipients demonstrated a marked increase in the relative proportion of the Firmicutes class Erysipelotrichia. These findings suggest that transplantation of gut-microbiomes can confer phenotypic responses to a drug and that the microbiome may be playing a major role in sympathomimetic-mediated hyperthermia. Abbreviations: 3,4-methylenedioxymethamphetamine (MDMA); methylone-induced hyperthermic tolerant (MHT); methylone-naïve (MN); fecal microbiota transplantation (FMT); uncoupling protein (UCP); subcutaneous (sc); intraperitoneal (ip); brown adipose tissue (BAT); skeletal muscle (SKM); sympathetic nervous system (SNS); norepinephrine (NE); quantitative PCR (qRT-PCR); quantification cycle (Cq); High Performance Liquid Chromatography-Electrochemical Detection (HPLC-EC); amplicon sequence variants (ASVs); principal coordinates analysis (PCoA); permutational multivariate analysis (PERMANOVA).

Sirtuin 5 levels are limiting in preserving cardiac function and suppressing fibrosis in response to pressure overload.

Heart failure (HF) is the inability of the heart to pump blood sufficiently to meet the metabolic demands of the body. HF with reduced systolic function is characterized by cardiac hypertrophy, ventricular fibrosis and remodeling, and decreased cardiac contractility, leading to cardiac functional impairment and death. Transverse aortic constriction (TAC) is a well-established model for inducing hypertrophy and HF in rodents. Mice globally deficient in sirtuin 5 (SIRT5), a NAD+-dependent deacylase, are hypersensitive to cardiac stress and display increased mortality after TAC. Prior studies assessing SIRT5 functions in the heart have all employed loss-of-function approaches. In this study, we generated SIRT5 overexpressing (SIRT5OE) mice, and evaluated their response to chronic pressure overload using TAC. Compared to littermate controls, SIRT5OE mice were protected against adverse functional consequences of TAC, left ventricular dilation and impaired ejection fraction. Transcriptomic analysis revealed that SIRT5 suppresses key HF sequelae, including the metabolic switch from fatty acid oxidation to glycolysis, immune activation, and fibrotic signaling pathways. We conclude that SIRT5 is a limiting factor in the preservation of cardiac function in response to experimental pressure overload.

Effort-reward balance and work motivation in rats: Effects of context and order of experience.

Being motivated means exerting effort toward a goal. The 'law of least work' emphasizes a preference for exerting relatively less effort. The law crosses boundaries among species and between physical and mental work. Organisms should be highly sensitive to shifts in effort-reward balance (ERB) in order to make optimal choices. We used a free operant-foraging task to investigate changes in ERB on choice between options requiring more or less effort. Results showed a consistent preference for the option with less effort and insensitivity to shifts in ERB. A second aim explored the influence of order of experience on effort choice. Choice for the more effortful option significantly increased after experiencing an equal effort-reward relationship during the initial free operant-foraging session. This relative increase in choice for the effortful option persisted even after effort-reward imbalance. The findings highlight the importance of contextual factors such as order of experience when examining the impact of shifting effort-reward associations. Instead of ignoring or reducing order effects, the sequence of experience (e.g. for shifts in ERB) could be manipulated to enhance or reduce value of outcomes or effort itself.

Understanding sciatica: illness and treatment beliefs in a lumbar radicular pain population. A qualitative interview study.

BACKGROUND: Several pathological processes contribute to lumbar radicular pain (LRP), commonly known as sciatica. It is not known how patients rationalise the experience of sciatica or understand the diagnosis. Providing clinicians with a better understanding of how patients conceptualise sciatica will help them to tailor information for patients on the management and treatment of the condition. AIM: To understand patients' beliefs regarding their illness following a diagnosis of LRP, how these beliefs were developed, and the impact of illness beliefs on treatment beliefs. DESIGN & SETTING: Qualitative interview study from a single NHS musculoskeletal interface service (in Wales, UK). METHOD: Thirteen patients recently diagnosed with LRP were consecutively recruited. Individual semi-structured interviews were recorded and transcribed. Data were analysed using a thematic approach. RESULTS: Four main themes were generated: (1) the illness experience (2) the concept of sciatica, (3) treatment beliefs, and (4) the desire for credible information. CONCLUSION: The diagnosis of LRP is often communicated and understood within a compressive conceptual illness identity. Explaining symptoms with a compressive pathological model is easily understood by patients but may not accurately reflect the spectrum of pathological processes known to contribute to radicular pain. This model appears to inform patient beliefs about treatments. Clinicians should take care to fully explain the pathology prior to shared decision-making with patients.

Gender differences in tolerance to the hyperthermia mediated by the synthetic cathinone methylone.

The toxidrome associated with death from the synthetic cathinones includes hyperthermia as part of the sympathomimetic syndrome. Here, we examine the gender differences in the development of tolerance to the hyperthermia mediated by the synthetic cathinone methylone. In addition to temperature changes, expression differences in genes encoding the uncoupling proteins (UCP) 1 & 3, and TGR5 in skeletal muscle (SKM) and brown adipose tissue (BAT) were examined. Male and female rats were treated weekly with methylone (10 mg/kg). The females developed a tolerance to the methylone-induced hyperthermia by week two of drug exposure. By the third week, females displayed a hypothermic response to methylone. Conversely, males continued to display a hyperthermic response up to and including week four. At week four, the males demonstrated a significantly lower hyperthermia and a complete tolerance seen at week five with no significant hyperthermia. Tissue samples collected after treatment on the sixth week indicate that chronic exposure to methylone reduced UCP1 expression in SKM and BAT of the female rats. Only the females displayed increased TGR5 expression in BAT. UCP3 expression increased in both the SKM and BAT of the males and females. The differences between responses in male and female subjects further demonstrate the need for gender studies in the toxicology associated with drugs with abuse potential.

Systematic review of the measurement properties of self-report physical activity questionnaires in healthy adult populations.

OBJECTIVE: This systematic review evaluated the measurement properties of current self-report physical activity questionnaires (SRPAQs) completed within healthy adult populations. DESIGN: Two reviewers independently searched seven electronic databases and hand searched for articles investigating measurement properties of a SRPAQ evaluating physical activity over the previous 6 months. Articles published from 1 May 2001 to 4 December 2014 were systematically screened and eligible studies were not limited to English language sources. Articles investigating specific race, gender or socioeconomic populations were excluded. RESULTS: 10 studies investigating 10 SRPAQs were included. The methodological quality of the included studies was evaluated using COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN) and ranged from 'poor' to 'good'. The Recent Physical Activity Questionnaire, International Physical Activity Questionnaires and Physical Activity Assessment Tool demonstrated good/excellent test-retest reliability (intra-class coefficient (ICC)=0.76, p<0.0001; r=0.627-0.91; r=0.618, p<0.001, respectively), but variable criterion validity (r=0.67, p<0.0001; r=-0.02-0.43; r=0.392, p<0.01, respectively). The single-item measure showed significant criterion validity against an accelerometer (for moderate to vigorous physical activity (MVPA) k=0.23, 95% CI 0.05 to 0.41; and physical activity ≥10 min bouts 0.39 (95% CI 0.14 to 0.64). Construct validity of the six-point scale and Human Activity Profile varied significantly with age, marital status and presence of comorbidities (p<0.05, <0.01, <0.000 and p<0.05, <0.05, <0.000, respectively). The 1 week Godlin-Shephard recall demonstrated 'moderate' validity with the gold standard measure of accelerometry (r=0.43). CONCLUSIONS: Inconclusive evidence exists. Further investigation of criterion validity of the short-form International Physical Activity Questionnaire is required, as it demonstrated excellent test-retest reliability. PROSPERO NUMBER: CRD42012002484.

Medical marijuana in the workplace: challenges and management options for occupational physicians.

Although possession and use of marijuana is prohibited by federal law, legalization in four states (Alaska, Colorado, Oregon, and Washington) and allowance for palliation and therapy in 19 others may reposition the drug away from the fringes of society. This evolving legal environment, and growing scientific evidence of its effectiveness for select health conditions, requires assessment of the safety and appropriateness of marijuana within the American workforce. Although studies have suggested that marijuana may be used with reasonable safety in some controlled environments, there are potential consequences to its use that necessitate employer scrutiny and concern. Several drug characteristics must be considered, including Δ-tetrahydrocannabinol (Δ-THC, or THC) concentration, route of administration, dose and frequency, and pharmacokinetics, as well as the risks inherent to particular workplace environments.

Thinking inside the box: the health cube paradigm for health and wellness program evaluation and design.

Appropriately constructed health promotions can improve population health. The authors developed a practical model for designing, evaluating, and improving initiatives to provide optimal value. Three independent model dimensions (impact, engagement, and sustainability) and the resultant three-dimensional paradigm were described using hypothetical case studies, including a walking challenge, a health risk assessment survey, and an individual condition management program. The 3-dimensional model is illustrated and the dimensions are defined. Calculation of a 3-dimensional score for program comparisons, refinements, and measurement is explained. Program 1, the walking challenge, had high engagement and impact, but limited sustainability. Program 2, the health risk assessment survey, had high engagement and sustainability but limited impact. Program 3, the on-site condition management program, had measurable impact and sustainability but limited engagement, because of a lack of program capacity. Each initiative, though successful in 2 dimensions, lacked sufficient evolution along the third axis for optimal value. Calculation of a 3-dimensional score is useful for health promotion program development comparison and refinements, and overall measurement of program success.

Cold hyperalgesia as a prognostic factor in whiplash associated disorders: a systematic review.

OBJECTIVE: To review and critically evaluate the existing literature for the prognostic value of cold hyperalgesia in Whiplash Associated Disorders (WAD). METHODS: Embase, PsycINFO, and Medline databases were systematically searched (from inception to 20th September 2011) for prospective studies investigating a prognostic ability for cold hyperalgesia in WAD. Reference lists and lead authors were cross-referenced. Two independent reviewers selected studies, and consensus was achieved via a third reviewer. The risk of bias in identified studies was systematically evaluated by two reviewers using previously published guidance. The influences of seven potential covariates of cold hyperalgesia were considered. Quantitative synthesis was planned and homogeneity assessed. A modified Grades of Recommendation, Assessment, Development, and Evaluation (GRADE) approach was used to qualitatively assess trials. RESULTS: The review screened 445 abstracts, from these 20 full text studies were retrieved and assessed for eligibility. Six prospective studies on four cohorts were identified and reviewed. Findings from all four cohorts supported cold hyperalgesia as a prognostic factor in WAD. CONCLUSIONS: There is moderate evidence supporting cold hyperalgesia as a prognostic factor for long-term pain and disability outcome in WAD. Further validation of the strength of this relationship and the influence of covariates are required. The mechanism for this relationship is unknown.

The Stress Strategy Test in relation to personality traits and cognitive abilities.

Relations between results of the Stress Strategy Test (SST), a computerized test the first author developed partly involving the Stroop effect, and of an interview and an intelligence test used by the Swedish armed forces as selection instruments for placement of recruits in type of military service were investigated in a group of 3,205 recruits. The SST results were found able to explain 17.64% and 12.25% of the variance in results of the intelligence test and the interview, respectively. Emotional stability and psychological energy were the two interview variables most closely related to results of the SST. The usefulness of the SST in a selection context is discussed.

Toothbrush bristle wear and adherence of Streptococcus mutans.

PURPOSE: The purpose of this study was conducted to determine if bristle wear impacts the adherence of Streptococcus mutans on toothbrushes and to evaluate whether it affects the extent of adherence at 0, 8, and 24 hours after air-drying. METHODS: Sixty toothbrushes--composed of 20 each from 3 different groups and defined by brand, brush trim, and head shape--were used in this study. Bristle wear on half of the toothbrushes was achieved using an orthodontic typodont with metal bonds and brackets and evaluated by 4 independent observers. New and worn toothbrushes were inoculated with 5 mutans, rinsed in tap water, and air-dried for 0, 8, and 24 hours. Four tufts were removed from the brush heads at each time point, placed in saline and vortexed to remove bacteria. Bacteria were aerobically grown on Mitis Salivarius Agar plates until colony-forming units could be counted. RESULTS: The toothbrush group impacts adherence of 5 mutans on both new and worn toothbrushes at 0, 8, and 24 hours after air-drying, with new toothbrushes harboring significantly more S mutans than worn toothbrushes at 0 hours. CONCLUSIONS: The results have implications for the design of toothbrush tufts as well as storage of toothbrushes in the home.

Investigating a computerised test of the Stroop effect extended by inclusion of a third, more difficult task.

A validation study of a computerised test recently developed involving the Stroop effect, extended here by inclusion of a third, more difficult test series, is presented. Three groups of men belonging to the Swedish armed forces and adjudged to differ in their qualifications (20, 32, and 19 men of levels 1, 2, and 3, respectively) and a fourth group of 18 men convicted of serious crimes of violence were given this test, termed the Stress Strategy Test. Discriminant analysis of the test's 12 variables (four for each of the three test series) yielded a discriminant power of 65% for the total group, highest for the level 1 group (80%) and for the nonmilitary group (72%), results substantially better than obtained for the original version of the test with use of similar subject groups.

The stroop strategy test: investigation of a computerised version of the Stroop test for classifying Swedish military servicemen.

A computerised test termed the Stroop Strategy Test, utilising the Stroop effect, is described. To assess the test's usefulness and discriminant power, it was given to three military groups adjudged on the basis of interview and a test of intelligence to differ in their qualifications (61 men of Level 1, 41 of Level 2, and 17 of Level 3, in descending order) and a group of 16 men imprisoned or put in detention school for violent behaviour. In a discriminant analysis in which the eight measures the test provided were included and the four groups were compared yielded a discriminant power of 52.6% for the group as a whole, highest (56%) for the military Level 1 group. The potential usefulness of the test is discussed.