PPARG activation promotes the proliferation of colorectal cancer cell lines and enhances the antiproliferative effect of 5-fluorouracil.

BACKGROUND: Peroxisome proliferator-activated receptor gamma (PPARG) is a member of the nuclear receptor family. It is involved in the regulation of adipogenesis, lipid metabolism, insulin sensitivity, vascular homeostasis and inflammation. In addition, PPARG agonists, known as thiazolidinediones, are well established in the treatment of type 2 diabetes mellitus. PPARGs role in cancer is a matter of debate, as pro- and anti-tumour properties have been described in various tumour entities. Currently, the specific role of PPARG in patients with colorectal cancer (CRC) is not fully understood. MATERIAL AND METHODS: The prognostic impact of PPARG expression was investigated by immunohistochemistry in a case-control study using a matched pair selection of CRC tumours (n = 246) with either distant metastases to the liver (n = 82), lung (n = 82) or without distant metastases (n = 82). Its effect on proliferation as well as the sensitivity to the chemotherapeutic drug 5-fluorouracil (5-FU) was examined after activation, inhibition, and transient gene knockdown of PPARG in the CRC cell lines SW403 and HT29. RESULTS: High PPARG expression was significantly associated with pulmonary metastasis (p = 0.019). Patients without distant metastases had a significantly longer overall survival with low PPARG expression in their tumours compared to patients with high PPARG expression (p = 0.045). In the pulmonary metastasis cohort instead, a trend towards longer survival was observed for patients with high PPARG expression in their tumour (p = 0.059). Activation of PPARG by pioglitazone and rosiglitazone resulted in a significant dose-dependent increase in proliferation of CRC cell lines. Inhibition of PPARG by its specific inhibitor GW9662 and siRNA-mediated knockdown of PPARG significantly decreased proliferation. Activating PPARG significantly increased the CRC cell lines sensitivity to 5-FU while its inhibition decreased it. CONCLUSION: The prognostic effect of PPARG expression depends on the metastasis localization in advanced CRC patients. Activation of PPARG increased malignancy associated traits such as proliferation in CRC cell lines but also increases sensitivity towards the chemotherapeutic agent 5-FU. Based on this finding, a combination therapy of PPARG agonists and 5-FU-based chemotherapy constitutes a promising strategy which should be further investigated.

Muscle wobbling mass dynamics: eigenfrequency dependencies on activity, impact strength, and ground material.

In legged locomotion, muscles undergo damped oscillations in response to the leg contacting the ground (an impact). How muscle oscillates varies depending on the impact situation. We used a custom-made frame in which we clamped an isolated rat muscle (M. gastrocnemius medialis and lateralis: GAS) and dropped it from three different heights and onto two different ground materials. In fully activated GAS, the dominant eigenfrequencies were 163 Hz, 265 Hz, and 399 Hz, which were signficantly higher (p < 0.05) compared to the dominant eigenfrequencies in passive GAS: 139 Hz, 215 Hz, and 286 Hz. In general, neither changing the falling height nor ground material led to any significant eigenfrequency changes in active nor passive GAS, respectively. To trace the eigenfrequency values back to GAS stiffness values, we developed a 3DoF model. The model-predicted GAS muscle eigenfrequencies matched well with the experimental values and deviated by - 3.8%, 9.0%, and 4.3% from the passive GAS eigenfrequencies and by - 1.8%, 13.3%, and - 1.5% from the active GAS eigenfrequencies. Differences between the frequencies found for active and passive muscle impact situations are dominantly due to the attachment of myosin heads to actin.

Corrigendum: Where have the dead gone?

[This corrects the article DOI: 10.3389/fmed.2022.837287.].

Training doctoral students in critical thinking and experimental design using problem-based learning.

BACKGROUND: Traditionally, doctoral student education in the biomedical sciences relies on didactic coursework to build a foundation of scientific knowledge and an apprenticeship model of training in the laboratory of an established investigator. Recent recommendations for revision of graduate training include the utilization of graduate student competencies to assess progress and the introduction of novel curricula focused on development of skills, rather than accumulation of facts. Evidence demonstrates that active learning approaches are effective. Several facets of active learning are components of problem-based learning (PBL), which is a teaching modality where student learning is self-directed toward solving problems in a relevant context. These concepts were combined and incorporated in creating a new introductory graduate course designed to develop scientific skills (student competencies) in matriculating doctoral students using a PBL format. METHODS: Evaluation of course effectiveness was measured using the principals of the Kirkpatrick Four Level Model of Evaluation. At the end of each course offering, students completed evaluation surveys on the course and instructors to assess their perceptions of training effectiveness. Pre- and post-tests assessing students' proficiency in experimental design were used to measure student learning. RESULTS: The analysis of the outcomes of the course suggests the training is effective in improving experimental design. The course was well received by the students as measured by student evaluations (Kirkpatrick Model Level 1). Improved scores on post-tests indicate that the students learned from the experience (Kirkpatrick Model Level 2). A template is provided for the implementation of similar courses at other institutions. CONCLUSIONS: This problem-based learning course appears effective in training newly matriculated graduate students in the required skills for designing experiments to test specific hypotheses, enhancing student preparation prior to initiation of their dissertation research.

Case of a Patient With Pancreatic Cancer With Sporadic Microsatellite Instability Associated With a BRAF Fusion Achieving Excellent Response to Immunotherapy.

In this case report, we discuss a case of pancreatic cancer bearing a BRAF fusion, leading to MAPK activation, MLHph, and finally MSI.

Indirect Antioxidant Effects of the Nitrite Anion: Focus on Xanthine Oxidase.

One electron reduction of nitrite (NO2 -) has been determined to be a significant, noncanonical source of nitric oxide (NO) with molybdopterin enzymes being identified as critical to this process. Of the molybdopterin enzymes identified as NO2 - reductases, xanthine oxidoreductase (XOR) is the most extensively studied. Paradoxically, XOR generates oxidants and thus can contribute to oxidative stress under inflammatory conditions when the oxidase form (XO) of XOR is abundant. However, under similar inflammatory conditions XO has been associated with NO generation, especially when NO2 - levels are elevated which begs the question: if reaction of nitrite with XO consumes electrons, then does it subsequently reduce oxidant generation? To address this question, electron paramagnetic resonance (EPR) was used, under controlled O2 tensions, to assess superoxide (O2 •-) generation by endothelial-bound XO plus xanthine and the resultant impact of introducing NO2 -. Nitrite diminished XO-derived O2 •- under hypoxia (1% O2) whereas at 21% O2, it had no impact. To confirm these results and discount contributions from the reaction of NO with O2 •-, molecular O2 consumption was assessed. The presence of NO2 - decreased the rate of XO/xanthine-dependent O2 consumption in a concentration-dependent manner with greater impact under hypoxic conditions (1% O2) compared to 21% O2. In a more biologic setting, NO2 - also diminished XO-dependent H2O2 formation in murine liver homogenates supplemented with xanthine. Interestingly, nitrate (NO3 -) did not alter XO-dependent O2 consumption at either 21% or 1% O2; yet it did slightly impact nitrite-mediated effects when present at 2:1 ratio vs. NO2 -. When combined, these data: 1) show a significant indirect antioxidant function for NO2 - by decreasing oxidant generation from XO, 2) demonstrate that both XO-derived H2O2 and O2 •- production are diminished by the presence of NO2 - and 3) incentivize further exploration of the difference between XO reaction with NO2 - vs. NO3 -.

Sponge EEG is equivalent regarding signal quality, but faster than routine EEG.

Objective: Emergency diagnostics, such as acquisition of an electroencephalogram (EEG), are of great diagnostic importance, but there is often a lack of experienced personnel. Wet active electrode sponge-based electroencephalogram (sp-EEG) systems can be applied rapidly and by inexperienced personnel. This makes them an attractive alternative to routine EEG (r-EEG) systems in these settings. Here, we examined the feasibility and signal quality of sp-EEG compared to r-EEG. Methods: In this case-control, single-blind, non-randomized study, EEG recordings using a sp- and a r-EEG system were performed in 18 individuals with a variety of epileptiform discharges and 11 healthy control subjects. The time was stopped until all electrodes in both systems displayed adequate skin-electrode impedances. The resulting 58 EEGs were visually inspected by 7 experienced, blinded neurologists. Raters were asked to score physiological and pathological graphoelements, and to distinguish between the different systems by visual inspection of the EEGs. Results: Time to signal acquisition for sp-EEG was significantly faster (4.8 min (SD 2.01) vs. r-EEG 13.3 min (SD 2.72), p < 0.001). All physiological and pathological graphoelements of all 58 EEGs could be identified. Raters were unable to distinguish between sp-EEG or r-EEG based on visual inspection of the EEGs alone. Conclusions: Sp-EEG represents a feasible alternative to r-EEG in emergency diagnostics or resource-limited settings. Significance: Given shortage of trained personnel or resources, the easy implementation and comparable quality of a novel sp-EEG system may increase general availability of EEG and thus improve patient care.

Clinical Impact of Structured Post-Operative Surveillance in Resected Pancreatic Adenocarcinoma: Results from a Retrospective Cohort Study.

INTRODUCTION: To this date, surgery remains the only potentially curative approach in the treatment of pancreatic cancer. To analyse the clinical impact of a structured post-operative follow-up programme, we retrospectively analysed a cohort of resected pancreatic adenocarcinoma patients treated at LMU Munich. METHODS: Pancreatic adenocarcinoma patients who underwent resection and presented for regular follow-up visits at our centre between 2002 and 2017 were identified from two existing study cohorts. Diagnosis of recurrences was categorised by timing (within or outside a scheduled follow-up visit) and detection modality (imaging, CA 19-9 increase, or clinical deterioration) and correlated with disease-free survival and overall survival (OS). RESULTS: One hundred and twenty-five patients with resected pancreatic adenocarcinoma were included in this analysis. Median OS in the whole cohort was 21.1 months. Of these 125 patients, 103 (82.4%) patients had a documented relapse. Tumour recurrences detected within a scheduled follow-up visit (n = 86, 83.5%) compared to recurrences becoming apparent at an unplanned visit (n = 17, 16.5%) were associated with a significantly improved OS (median 25.5 vs. 20.2 months, p = 0.019). Compared to patients with recurrence detected by clinical deterioration (n = 4, 3.9%), patients with recurrences detected by imaging or laboratory abnormalities (n = 99, 96.0%) had a longer median OS (24.8 vs. 15.1 months, p = 0.007). DISCUSSION: A structured follow-up after pancreatic ductal adenocarcinoma resection may have an impact on patient outcome. Prospective trials are needed to evaluate the clinical impact of post-operative follow-up programmes.

Bcl-xL as prognostic marker and potential therapeutic target in cholangiocarcinoma.

Intrahepatic, perihilar, and distal cholangiocarcinoma (iCCA, pCCA, dCCA) are highly malignant tumours with increasing mortality rates due to therapy resistances. Among the mechanisms mediating resistance, overexpression of anti-apoptotic Bcl-2 proteins (Bcl-2, Bcl-xL , Mcl-1) is particularly important. In this study, we investigated whether antiapoptotic protein patterns are prognostically relevant and potential therapeutic targets in CCA. Bcl-2 proteins were analysed in a pan-cancer cohort from the NCT/DKFZ/DKTK MASTER registry trial (n = 1140, CCA n = 72) via RNA-sequencing and transcriptome-based protein activity interference revealing high ranks of CCA for Bcl-xL and Mcl-1. Expression of Bcl-xL , Mcl-1, and Bcl-2 was assessed in human CCA tissue and cell lines compared with cholangiocytes by immunohistochemistry, immunoblotting, and quantitative-RT-PCR. Immunohistochemistry confirmed the upregulation of Bcl-xL and Mcl-1 in iCCA tissues. Cell death of CCA cell lines upon treatment with specific small molecule inhibitors of Bcl-xL (Wehi-539), of Mcl-1 (S63845), and Bcl-2 (ABT-199), either alone, in combination with each other or together with chemotherapeutics was assessed by flow cytometry. Targeting Bcl-xL induced cell death and augmented the effect of chemotherapy in CCA cells. Combined inhibition of Bcl-xL and Mcl-1 led to a synergistic increase in cell death in CCA cell lines. Correlation between Bcl-2 protein expression and survival was analysed within three independent patient cohorts from cancer centers in Germany comprising 656 CCA cases indicating a prognostic value of Bcl-xL in CCA depending on the CCA subtype. Collectively, these observations identify Bcl-xL as a key protein in cell death resistance of CCA and may pave the way for clinical application.

Muscle active force-length curve explained by an electrophysical model of interfilament spacing.

The active isometric force-length relation (FLR) of striated muscle sarcomeres is central to understanding and modeling muscle function. The mechanistic basis of the descending arm of the FLR is well explained by the decreasing thin:thick filament overlap that occurs at long sarcomere lengths. The mechanistic basis of the ascending arm of the FLR (the decrease in force that occurs at short sarcomere lengths), alternatively, has never been well explained. Because muscle is a constant-volume system, interfilament lattice distances must increase as sarcomere length shortens. This increase would decrease thin and thick-filament electrostatic interactions independently of thin:thick filament overlap. To examine this effect, we present here a fundamental, physics-based model of the sarcomere that includes filament molecular properties, calcium binding, sarcomere geometry including both thin:thick filament overlap and interfilament radial distance, and electrostatics. The model gives extremely good fits to existing FLR data from a large number of different muscles across their entire range of measured activity levels, with the optimized parameter values in all cases lying within anatomically and physically reasonable ranges. A local first-order sensitivity analysis (varying individual parameters while holding the values of all others constant) shows that model output is most sensitive to a subset of model parameters, most of which are related to sarcomere geometry, with model output being most sensitive to interfilament radial distance. This conclusion is supported by re-running the fits with only this parameter subset being allowed to vary, which increases fit errors only moderately. These results show that the model well reproduces existing experimental data, and indicate that changes in interfilament spacing play as central a role as changes in filament overlap in determining the FLR, particularly on its ascending arm.

The short term influence of right ventricular pacing burden on echocardiographic and spiroergometric parameters in patients with preserved left ventricular ejection fraction.

BACKGROUND: The incidence of worsened clinical outcome due to high right ventricular (RV) pacing burden in patients with preserved left ventricular function remains controversial. OBJECTIVE: To investigate the impact of RV pacing on several echocardiographic and spiroergometric parameters. METHODS: In 60 pacemaker patients with preserved left ventricular ejection fraction (LVEF) serial echocardiographies and spiroergometries were performed over a time course of 12 months. Additionally, in 48 patients retrospective echocardiographic analyses of the LV- and RV function were carried out up to 24 months after pacemaker implantation. RESULTS: The patients were divided into two groups: The high RV pacing burden group (hRVP: ≥ 40%) and the low RV pacing group (lRVP < 40%) according to the definitions in previous randomized MOST and DAVID trials. After a period of 12-month pacemaker therapy no changes to left ventricular end diastolic diameter (LVEDD), left ventricular end systolic diameter (LVESD), LVEF, E/A-ratio; E/E'-ratio and tricuspid annular plane systolic excursion (TAPSE) could be revealed, independently of the RV pacing burden. Additionally, after 24-month long term follow-up there were no differences in LVEF and TAPSE in both groups. Accordingly, no relevant changes of peak exercise capacity, ventilatory anaerobic threshold or maximal oxygen consumption could be demonstrated independently of the RV pacing. CONCLUSIONS: In pacemaker patients with preserved LVEF the burden of RV pacing has no adverse influence on several echocardiographic and spiroergometric surrogate parameters of pacemaker-induced cardiomyopathy after a follow-up of 12 to 24 month. Despite this, screening for pacemaker induced cardiomyopathy should be performed especially in the presence of new heart failure symptoms.

Foot function enabled by human walking dynamics.

Bipedal walking, the habitual gait for man, is rather unique in nature and poses particular challenges for balance and propulsion. The characteristic double-humped ground reaction force profile has been widely observed but not put into functional context. We propose a mathematical model that captures the dynamics of the human foot in walking including the characteristic motion of the center of pressure. Using this model, we analyze the functional interplay of all essential biomechanical contributors to foot dynamics in walking. Our results demonstrate the intricate interplay of a self-stabilizing mechanism which allows extending a leg's stance phase while simultaneously powering rapid swing by condensing the essentials of foot dynamics into a reductionist, biomechanical model. A theory is presented which identifies the foot to be the key functional element and which explains the global dynamics of human walking. The provided insights will impact gait therapy and rehabilitation, the development of assistive devices, such as leg prostheses and exoskeletons, and provide guidelines for the design and control of versatile humanoid robots.

Cross-bridge mechanics estimated from skeletal muscles' work-loop responses to impacts in legged locomotion.

Legged locomotion has evolved as the most common form of terrestrial locomotion. When the leg makes contact with a solid surface, muscles absorb some of the shock-wave accelerations (impacts) that propagate through the body. We built a custom-made frame to which we fixated a rat (Rattus norvegicus, Wistar) muscle (m. gastrocnemius medialis and lateralis: GAS) for emulating an impact. We found that the fibre material of the muscle dissipates between 3.5 and [Formula: see text] ranging from fresh, fully active to passive muscle material, respectively. Accordingly, the corresponding dissipated energy in a half-sarcomere ranges between 10.4 and [Formula: see text], respectively. At maximum activity, a single cross-bridge would, thus, dissipate 0.6% of the mechanical work available per ATP split per impact, and up to 16% energy in common, submaximal, activities. We also found the cross-bridge stiffness as low as [Formula: see text], which can be explained by the Coulomb-actuating cross-bridge part dominating the sarcomere stiffness. Results of the study provide a deeper understanding of contractile dynamics during early ground contact in bouncy gait.

Enantioselective Construction of Modular and Asymmetric Baskets.

The precise positioning of functional groups about the inner space of abiotic hosts is a challenging task and of interest for developing more effective receptors and catalysts akin to those found in nature. To address it, we herein report a synthetic methodology for preparing basket-like cavitands comprised of three different aromatics as side arms with orthogonal esters at the rim for further functionalization. First, enantioenriched A (borochloronorbornene), B (iodobromonorbornene), and C (boronorbornene) building blocks were obtained by stereoselective syntheses. Second, consecutive A-to-B and then AB-to-C Suzuki-Miyaura (SM) couplings were optimized to give enantioenriched ABC cavitand as the principal product. The robust synthetic protocol allowed us to prepare (a) an enantioenriched basket with three benzene sides and each holding either tBu, Et, or Me esters, (b) both enantiomers of a so-called "spiral staircase" basket with benzene, naphthalene, and anthracene groups surrounding the inner space, and (c) a photo-responsive basket bearing one anthracene and two benzene arms.

Rules of nature's Formula Run: Muscle mechanics during late stance is the key to explaining maximum running speed.

The maximum running speed of legged animals is one evident factor for evolutionary selection-for predators and prey. Therefore, it has been studied across the entire size range of animals, from the smallest mites to the largest elephants, and even beyond to extinct dinosaurs. A recent analysis of the relation between animal mass (size) and maximum running speed showed that there seems to be an optimal range of body masses in which the highest terrestrial running speeds occur. However, the conclusion drawn from that analysis-namely, that maximum speed is limited by the fatigue of white muscle fibres in the acceleration of the body mass to some theoretically possible maximum speed-was based on coarse reasoning on metabolic grounds, which neglected important biomechanical factors and basic muscle-metabolic parameters. Here, we propose a generic biomechanical model to investigate the allometry of the maximum speed of legged running. The model incorporates biomechanically important concepts: the ground reaction force being counteracted by air drag, the leg with its gearing of both a muscle into a leg length change and the muscle into the ground reaction force, as well as the maximum muscle contraction velocity, which includes muscle-tendon dynamics, and the muscle inertia-with all of them scaling with body mass. Put together, these concepts' characteristics and their interactions provide a mechanistic explanation for the allometry of maximum legged running speed. This accompanies the offering of an explanation for the empirically found, overall maximum in speed: In animals bigger than a cheetah or pronghorn, the time that any leg-extending muscle needs to settle, starting from being isometric at about midstance, at the concentric contraction speed required for running at highest speeds becomes too long to be attainable within the time period of a leg moving from midstance to lift-off. Based on our biomechanical model, we, thus, suggest considering the overall speed maximum to indicate muscle inertia being functionally significant in animal locomotion. Furthermore, the model renders possible insights into biological design principles such as differences in the leg concept between cats and spiders, and the relevance of multi-leg (mammals: four, insects: six, spiders: eight) body designs and emerging gaits. Moreover, we expose a completely new consideration regarding the muscles' metabolic energy consumption, both during acceleration to maximum speed and in steady-state locomotion.

A geometry- and muscle-based control architecture for synthesising biological movement.

A key problem for biological motor control is to establish a link between an idea of a movement and the generation of a set of muscle-stimulating signals that lead to the movement execution. The number of signals to generate is thereby larger than the body's mechanical degrees of freedom in which the idea of the movement may be easily expressed, as the movement is actually executed in this space. A mathematical formulation that provides a solving link is presented in this paper in the form of a layered, hierarchical control architecture. It is meant to synthesise a wide range of complex three-dimensional muscle-driven movements. The control architecture consists of a 'conceptional layer', where the movement is planned, a 'structural layer', where the muscles are stimulated, and between both an additional 'transformational layer', where the muscle-joint redundancy is resolved. We demonstrate the operativeness by simulating human stance and squatting in a three-dimensional digital human model (DHM). The DHM considers 20 angular DoFs and 36 Hill-type muscle-tendon units (MTUs) and is exposed to gravity, while its feet contact the ground via reversible stick-slip interactions. The control architecture continuously stimulates all MTUs ('structural layer') based on a high-level, torque-based task formulation within its 'conceptional layer'. Desired states of joint angles (postural plan) are fed to two mid-level joint controllers in the 'transformational layer'. The 'transformational layer' communicates with the biophysical structures in the 'structural layer' by providing direct MTU stimulation contributions and further input signals for low-level MTU controllers. Thereby, the redundancy of the MTU stimulations with respect to the joint angles is resolved, i.e. a link between plan and execution is established, by exploiting some properties of the biophysical structures modelled. The resulting joint torques generated by the MTUs via their moment arms are fed back to the conceptional layer, closing the high-level control loop. Within our mathematical formulations of the Jacobian matrix-based layer transformations, we identify the crucial information for the redundancy solution to be the muscle moment arms, the stiffness relations of muscle and tendon tissue within the muscle model, and the length-stimulation relation of the muscle activation dynamics. The present control architecture allows the straightforward feeding of conceptional movement task formulations to MTUs. With this approach, the problem of movement planning is eased, as solely the mechanical system has to be considered in the conceptional plan.