Level Of Activation Research Articles

8343 The Role Of Pancreatic Nerve Activity On The Development Of Beta Cell Dysfunction With The Macrophage-Beta Cell Interaction

Abstract Disclosure: J. Yu: None. Y. Wang: None. L. Sha: None. Islet inflammation is a vital factor to the beta cell dysfunction which is driven by inflammatory cytokines TNF-α and IL-1β released by resident macrophages. Pancreatic nerve innervates islet function and our previous study suggested that pancreatic nerve activity might involved in the development of pancreatic beta cell dysfunction. However, it is not known whether pancreatic nerve activity modulates macrophage activity and contributes to the macrophage - beta cell interaction. The aim of our study was to investigate the role of nerve activity on it and the possible mechanism. Methods: Sprague Dawley Rats and high fat diet-induced obese rates were used. An intrapancreatic nerve truck with its intact innervated pancreatic islets were used for the study. The nerve activity was evoked with electric nerve stimulation and insulin and ATP released by beta cells, TNF-α released by macrophages, and IL-1β released mainly by macrophages were measured with nerve activation. Results: Insulin secretion was induced significantly with the nerve activation (8 Hz). With nerve activation, ATP level was significantly increased by 25.5 ± 11.7%. The increased releases of insulin and ATP were abolished with cholinergic M receptor blocker atropine. With nerve activation, TNF-α and IL-1β levels were increased by 25.3±7.6% and 51.8±11.7%, respectively (p<0.05). Interestingly, the increases of TNF-α and IL-1β with nerve activation were also blocked by atropine. Since it is known that acetylcholine does not induce pro-inflammatory cytokine release in macrophages, we suspected that the nerve activity induced TNF-α and IL-1β releases were indirect. ATP is likely a candidate for the indirect effect as receptors. With applying Suramin, a ATP receptor blocker, nerve activation increased TNF-α release was inhibited significantly and IL-1β release was trended to inhibited. atropine, an M receptor blocker, compared with the Control group, decreased by 88.0±3.7% and 63.1±19.6%, respectively (p< 0.01). In obese rats, nerve activation increased TNF-α and IL-1β content by 68.1 ± 22.6% and 39.7 ± 7.4%, respectively, and suramin significantly inhibited the nerve activation induced increase of TNF-α release and trended to inhibit IL-1β release. Conclusion: Our results suggested that the pancreatic nerve activity contribute to the macrophage-beta cell interaction in the development of beta cells dysfunction by enhancing ATP release from islet beta cells. Presentation: 6/3/2024

Alterations in magnitude and spatial distribution of erector spinae muscle activity in cyclists with a recent history of low back pain.

While cycling offers several health benefits, repetitive loading and maintenance of static postures for prolonged periods expose cyclists to low back pain (LBP). Despite high LBP prevalence in cyclists, underlying pathomechanics and specific lumbar region muscle activation patterns during cycling are unclear. Here, we compared lumbar erector spinae (ES) muscles activation and spatial distribution activity in cyclists with and without recent LBP history. Ten cyclists with recent LBP history (LBPG; Oswestry Disability Index score ~ 17.8%) and 11 healthy cyclists (CG) were recruited. After assessing the Functional Threshold Power (FTP), participants underwent an incremental cycling test with 4 × 3min steps at 70%, 80%, 90%, and 100% of their FTP. High-density surface electromyography (HDsEMG) signals were recorded from both lumbar ES using two 64-channel grids. Information about ES activation levels (root-mean-square, RMS), degree of homogeneity (entropy), and cranio-caudal displacement of muscle activity (Y-axis coordinate of the barycenter of RMS maps) was extracted from each grid separately and then grand-averaged across both grids. Repeated-measure 2-way ANOVAs showed a significant intensity by group interaction for RMS amplitude (p = 0.003), entropy (p = 0.038), and Y-bar displacement (p = 0.033). LBPG increased RMS amplitude between 70-100% (+ 19%, p = 0.010) and 80-100% FTP (+ 21%, p = 0.004) and decreased entropy between 70-100% FTP (-8.4%, p = 0.003) and 80-100% FTP (-8.5%, p = 0.002). Between-group differences emerged only at 100% FTP (+ 9.6%, p = 0.049) for RMS amplitude. Our findings suggest that cyclists with recent LBP history exhibit higher ES muscles activation and less homogeneous activity compared to healthy controls, suggesting potential inefficient muscle recruitment strategy. HEC-DSB/09-2023.

C5a Induces Inflammatory Signaling and Apoptosis in PC12 Cells through C5aR-Dependent Signaling: A Potential Mechanism for Adrenal Damage in Sepsis.

The complement system is critically involved in the pathogenesis of sepsis. In particular, complement anaphylatoxin C5a is generated in excess during sepsis, leading to cellular dysfunction. Recent studies have shown that excessive C5a impairs adrenomedullary catecholamine production release and induces apoptosis in adrenomedullary cells. Currently, the mechanisms by which C5a impacts adrenal cell function are poorly understood. The PC12 cell model was used to examine the cellular effects following treatment with recombinant rat C5a. The levels of caspase activation and cell death, protein kinase signaling pathway activation, and changes in inflammatory protein expression were examined following treatment with C5a. There was an increase in apoptosis of PC12 cells following treatment with high-dose C5a. Ten inflammatory proteins, primarily involved in apoptosis, cell survival, and cell proliferation, were upregulated following treatment with high-dose C5a. Five inflammatory proteins, involved primarily in chemotaxis and anti-inflammatory functions, were downregulated. The ERK/MAPK, p38/MAPK, JNK/MAPK, and AKT protein kinase signaling pathways were upregulated in a C5aR-dependent manner. These results demonstrate an apoptotic effect and cellular signaling effect of high-dose C5a. Taken together, the overall data suggest that high levels of C5a may play a role in C5aR-dependent apoptosis of adrenal medullary cells in sepsis.

Clinicogenomic predictors of outcomes in patients with hepatocellular carcinoma treated with immunotherapy.

Immune checkpoint inhibitor (ICI) combinations extend overall survival (OS) while anti-PD-1/L1 monotherapy is non-inferior to sorafenib in treatment-naïve, patients with advanced hepatocellular carcinoma (HCC). Clinicogenomic features are posited to influence patient outcomes. The primary objective of this retrospective study was to define the clinical, pathologic, and genomic factors associated with outcomes to ICI therapy in patients with HCC. Patients with histologically confirmed advanced HCC treated with ICI at Memorial Sloan Kettering Cancer Center from 2012 to 2022 were included. Association between clinical, pathological, and genomic characteristics were assessed with univariable and multivariable Cox regression model for progression-free survival (PFS) and OS. Two-hundred and forty-two patients were treated with ICI-based therapy. Patients were predominantly male (82%) with virally mediated HCC (53%) and Child Pugh A score (70%). Median follow-up was 28 months (0.5-78.4). Median PFS for those treated in 1st line, 2nd line and ≥ 3rd line was 4.9 (range: 2.9-6.2), 3.1 (2.3-4.0), and 2.5 (2.1-4.0) months, respectively. Median OS for those treated in 1st line, 2nd line, and ≥ 3rd line was 16 (11-22), 7.5 (6.4-11), and 6.4 (4.6-26) months, respectively. Poor liver function and performance status associated with worse PFS and OS, while viral hepatitis C was associated with favorable outcome. Genetic alterations were not associated with outcomes. Clinicopathologic factors were the major determinates of outcomes for patients with advanced HCC treated with ICI. Molecular profiling did not aid in stratification of ICI outcomes. Future studies should explore alternative biomarkers such as the level of immune activation or the pretreatment composition of the immune tumor microenvironment.

Construction of self-driving anti-αFR CAR-engineered NK cells based on IFN-γ and TNF-α synergistically induced high expression of CXCL10

IntroductionOvarian cancer is the most malignant gynecological tumor. Previous studies have demonstrated that chimeric antigen receptor (CAR)-engineered NK-92 cells targeting folate receptor α (αFR) (NK-92-αFR-CAR) can specifically kill αFR-positive ovarian cancer cells. However, the migration barrier restricts antitumor effects of CAR-engineered cells. ObjectivesTo elucidate the mechanism by which NK-92-αFR-CAR cells induce the secretion of chemokine CXCL10 during killing ovarian cancer cells. It is speculated that NK-92-αFR-CAR-CXCR3A can target αFR and have chemotaxis of CXCL10, and they may have stronger killing effect of ovarian cancer. MethodsStudy the mechanism of CXCL10 expression strongly induced by TNF-α and IFN-γ combined stimulation in ovarian cancer cells. Construct the fourth generation of NK-92-αFR-CAR-CXCR3A cells, which were co-expressed CXCR3A and αFR-CAR. Evaluate the killing and migration effects of NK-92-αFR-CAR-CXCR3A in vitro and in vivo. ResultsRNA sequencing (RNA-seq) first revealed that the expression level of the chemokine CXCL10 was most significantly increased in ovarian cancer cells co-cultured with NK-92-αFR-CAR. Secondly, cytokine stimulation experiments confirmed that IFN-γ and TNF-α secreted by NK-92-αFR-CAR synergistically induced high CXCL10 expression in ovarian cancer cells. Further signaling pathway experiments showed that IFN-γ and TNF-α enhanced the activation level of the IFN-γ-IFNGR-JAK1/2-STAT1-CXCL10 signaling axis. Cytotoxicity experiments showed that NK-92-αFR-CAR-CXCR3A cells could not only efficiently kill αFR-positive ovarian cancer cells in vitro but also secrete IFN-γ and TNF-α. Higher migration than that of NK-92-αFR-CAR was detected in NK-92-αFR-CAR-CXCR3A using transwell assay. NK-92-αFR-CAR-CXCR3A effectively killed tumor cells in different mouse xenograft models of ovarian cancer and increased infiltration into tumor tissue. ConclusionThis study confirmed that IFN-γ and TNF-α secreted by αFR-CAR-engineered NK cells can synergistically induce high expression of CXCL10 in ovarian cancer cells and constructed self-driving αFR-CAR-engineered NK cells that can break through migration barriers based on CXCL10, which may provide a new therapeutic weapon for ovarian cancer.

CO oxidation on single Pt atom supported by two-dimensional ZnO monolayer: Reaction mechanism and precursor activation

CO oxidation is an effective solution for controlling CO emissions, and supported single-atom catalysts have shown excellent catalytic reactivity for chemical reactions. CO oxidation on a single Pt atom supported by a two-dimensional ZnO monolayer is investigated by combing ab initio molecular dynamics and density functional theory calculations. Seven reaction mechanisms are identified, including two new trimolecular Elie-Riedel mechanisms; six of them, except for the Mars-van Krevelen mechanism, have rate-limiting step energy barriers between 0.31 and 0.47 eV, which is lower than the experimental activation barrier of 0.81 eV on single Pt atoms supported by bulk ZnO, and is much lower than that of about 1.0 eV on Pt(111) surface. Furthermore, two preferred reaction mechanisms are discovered, an Eley-Rideal mechanism and a trimolecular Eley-Rideal mechanism, both of which have rate-limiting step energy barriers as low as 0.31 eV. Our results show that the more antibonding orbitals are filled in the reaction precursor, the higher its activation level. Activation of precursors plays an important role in CO oxidation and leads to different mechanisms. These results provide insights into the design of efficient single-atom catalysts supported by two-dimensional materials for the removal of CO pollutants.

TLR2 reprograms glucose metabolism in CD4+ T cells of rheumatoid arthritis patients to mediate cell hyperactivation and TNF-α secretion.

Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease in which activated CD4+ T cells participate in the disease process by inducing inflammation. We aimed to investigate the role of Toll-like receptor 2 (TLR2) on CD4+ T cells in RA patients, and to elucidate the underlying mechanisms by which TLR2 contributes to the pathogenesis of RA. Serum samples were collected from RA patients and healthy controls. Soluble TLR2 levels were quantified using an enzyme-linked immunosorbent assay (ELISA). Flow cytometry was employed to assess the TLR2 expression level, activation status, cytokine production, reactive oxygen species (ROS) levels, and glucose uptake capacity of CD4+ T cells. Quantitative polymerase chain reaction (qPCR) was used to measure the expression of enzymes associated with glucose and lipid metabolism. The concentration of lactic acid in the culture supernatant was determined using a dedicated detection kit. RA patients had higher levels of TLR2 in their serum, which positively correlated with C-reactive protein and rheumatoid factor. The expression level of TLR2 in CD4+ T cells of RA patients was increased, and TLR2+ cells showed higher activation levels than TLR2- cells. Activation of TLR2 in CD4+ T cells of RA patients promoted their activation, TNF-α secretion, and increased production of ROS. Furthermore, TLR2 activation led to changes in enzymes related to glucose metabolism, causing a shift in glucose metabolism towards the pentose phosphate pathway. Blocking oxidative phosphorylation and the pentose phosphate pathway had varying effects on CD4+ T cell function. TLR2 reprograms the glucose metabolism of CD4+ T cells in RA patients, contributing to the development of RA through ROS-mediated cell hyperactivation and TNF-α secretion. Key Points • TLR2 is upregulated in CD4+ T cells of RA patients and correlates with disease severity markers such as CRP and RF. • Activation of TLR2 in CD4+ T cells promotes cell activation, TNF-α secretion, and increased ROS production, contributing to the pathogenesis of RA. • TLR2 activates glucose metabolism in CD4+ T cells, shifting towards the pentose phosphate pathway, which may be a novel therapeutic target for RA treatment. • Blocking glucose metabolism and ROS production can reduce CD4 + T cell hyperactivation and TNF-α secretion, indicating potential therapeutic strategies for RA management.

Residual force enhancement decreases when scaling from the single muscle fiber to joint level in humans

BackgroundResidual force enhancement (rFE), defined as increased isometric force following active lengthening compared to a fixed-end isometric contraction at the same muscle length and level of activation, is present across all scales of muscle. While rFE is always present at the cellular level, often rFE “non-responders” are observed during joint-level voluntary contractions. MethodsWe compared rFE between the joint level and single fiber level (vastus lateralis biopsies) in 16 young males. In vivo voluntary knee-extensor rFE was measured by comparing steady-state isometric torque between a stretch-hold (maximal activation at 150°, stretch to 70°, hold) and a fixed-end isometric contraction, with ultrasonographic recording of vastus lateralis fascicle length (FL). Fixed-end contractions were performed at 67.5°, 70.0°, 72.5°, and 75.0°; the joint angle that most closely matched FL of the stretch-hold contraction's isometric steady-state was used to calculate rFE. The starting and ending FLs of the stretch-hold contraction were expressed as % optimal FL, determined via torque-angle relationship. ResultsIn single fiber experiments, the starting and ending fiber lengths were matched relative to optimal length determined from in vivo testing, yielding an average sarcomere excursion of ∼2.2–3.4µm. There was a greater magnitude of rFE at the single fiber (∼20%) than joint level (∼5%) (p = 0.004), with “non-responders” only observed at the joint level. ConclusionBy comparing rFE across scales within the same participants, we show the development of the rFE non-responder phenomenon is upstream of rFE's cellular mechanisms, with rFE only lost rather than gained when scaling from single fibers to the joint level.

Identification of NET formation and the renoprotective effect of degraded NETs in lupus nephritis.

To explore molecular biomarkers associated with the pathophysiology and therapy of lupus nephritis (LN), we conducted a joint analysis of transcriptomic data from 40 peripheral blood mononuclear cells (PBMCs) (GSE81622) and 21 kidney samples (GSE112943) from the Gene Expression Omnibus database using bioinformatics. A total of 976 and 2,427 differentially expressed genes (DEGs) were identified in PBMCs and renal tissues. Seven and two functional modules closely related to LN were identified. Further enrichment analysis revealed that the neutrophil activation pathway was highly active in both PBMCs and the kidney. Subsequently, 16 core genes closely associated with LN were verified by protein-protein interaction screening and quantitative PCR. In vitro cell models and MRL/lpr mouse models confirmed that the abnormal expression of these core genes was closely linked to neutrophil extracellular traps (NETs) generated by neutrophil activation, while degradation of NETs led to downregulation of core gene expression, thereby improving pathological symptoms of LN. Therefore, identification of patients with systemic lupus erythematosus exhibiting abnormal expression patterns for these core genes may serve as a useful indicator for kidney involvement. In addition, targeting neutrophils to modulate their activation levels and inhibit aberrant expression of these genes represents a potential therapeutic strategy for treating LN. NEW & NOTEWORTHY The mechanisms by which immune cells cause kidney injury in lupus nephritis are poorly understood. We integrated and analyzed the transcriptomic features of PBMCs and renal tissues from the GEO database to identify key molecular markers associated with neutrophil activation. We confirmed that neutrophil extracellular traps (NETs) formed by neutrophil activation promoted the upregulation of key genes in cell and animal models. Targeted degradation of NETs significantly ameliorated kidney injury in MRL/lpr mice.

Strategies for designing machine learning models in renewable energy with insufficient data

In the area of renewable energy including wind, solar, and electricity, Machine Learning has established a far-reaching conceptual and algorithmic design framework. However, the design of Machine Learning architectures typically relies on substantial amounts of data. The data acquisition process often involves complex tasks of gathering, cleaning, and labeling data. This challenge is exacerbated in the energy sector due to concerns related to data security and the uniqueness of development initiatives, leading to a potential scarcity of available data. Quite commonly, researchers encounter a vast array of innovative projects that have not been studied in the past and therefore only limited data become available. Therefore, there is an urgent and highly motivated need to develop machine learning strategies that can construct reliable models in the presence of limited data. In this study, three machine learning models tailored for small-scale data scenarios are developed and analyzed. The first model, a fuzzy cluster ensemble approach, focuses on utilizing small training samples treated as the core of clustering. By applying clustering algorithms, one derives membership functions reflecting the structure of data that facilitate accurate predictions. The second model, a kernel integration framework, combines kernel functions with limited training data to calculate activation levels, enabling accurate prediction outputs. For both models, the final predictions are generated by applying weighted aggregations, where the weights are derived from the respective membership grades and activation levels of the data. Additionally, a neural network contrastive learning model is introduced, which enhances the training process by employing pairwise comparisons among data points. This method effectively increases the amount of useable training data, improving the model's ability to make accurate predictions. Together, these approaches provide a robust solution for making reliable predictions even in data-scarce scenarios within renewable energy research. Moreover, the predictive performance of these models is quantified using the Root Mean Square Error criterion. This study also assesses the performance of the developed models by forming a granular characterization of results delivered by the designed models. A comprehensive suite of experimental studies is included, accompanied by an exhaustive comparative analysis. Compared to conventional machine learning models, especially neural networks, the designed models offer reliable results even with limited training data, showing an improvement of around 20 %, thus laying the groundwork for digitization processes in the realm of sustainable energy.

Implementing a multisite shared haemodialysis care programme.

Adults receiving centre-based haemodialysis (HD) have low levels of patient activation which are associated with poorer outcomes. Shared haemodialysis care (SHC) describes an intervention whereby individuals are supported to undertake elements of their treatment to improve their activation levels and promote better self-care. This project aimed to increase the proportion of those performing SHC in seven HD centres within the Oxford Kidney Unit's catchment area. Sequential Plan-Do-Study-Act (PDSA) cycles effected change first in two central HD centres, in cycles 1 and 2, before rolling out to five satellite HD centres, in cycles 3 and 4. Cycle 1 explored and transformed staff perceptions regarding SHC using a questionnaire and teaching sessions while in cycle 2, staff partnered with patients to develop leaflets and noticeboards to improve awareness and participation. These interventions were then rolled out to the remaining HD centres in PDSA cycles 3 and 4. Other interventions included: Enrolling staff and patients in virtual training courses; designating SHC 'Champions'; engagement with a national SHC forum; and changes to the electronic patient record to enable the monitoring of patient SHC opportunity and to promote sustainable change. Outcome measurement data on the number of patients performing SHC and the number at different defined stages of SHC competency were captured monthly. In April 2022, only 4% (19/483) of those receiving centre-based HD performed any aspect of SHC. By the end of the project in December 2023, this had increased to 43% (220/511). There was a significant and sustained growth in the stage of patient SHC competency as well as the number of patients performing SHC in each HD centre. The project demonstrated that it is possible to implement, scale-up and maintain a multisite SHC programme even with little baseline staff and patient SHC experience.

PPP3CB overexpression mediates EGFR TKI resistance in lung tumors via calcineurin/MEK/ERK signaling.

Despite initial high response rates to first-line EGFR TKI, all non-small-cell lung cancer (NSCLC) with EGFR-activating mutation will ultimately develop resistance to treatment. Identification of resistance mechanisms is critical to adapt treatment and improve patient outcomes. Here, we show that a PPP3CB transcript that encodes full-length catalytic subunit 2B of calcineurin accumulates in EGFR-mutant NSCLC cells with acquired resistance against different EGFR TKIs and in post-progression biopsies of NSCLC patients treated with EGFR TKIs. Neutralization of PPP3CB by siRNA or inactivation of calcineurin by cyclosporin A induces apoptosis in resistant cells treated with EGFR TKIs. Mechanistically, EGFR TKIs increase the cytosolic level of calcium and trigger activation of a calcineurin/MEK/ERK pathway that prevents apoptosis. Combining EGFR, calcineurin, and MEK inhibitors overcomes resistance to EGFR TKI in both in vitro and in vivo models. Our results identify PPP3CB overexpression as a new mechanism of acquired resistance to EGFR TKIs, and provide a promising therapeutic approach for NSCLC patients that progress under TKI treatment.

The double 90 rule: A new strategy for resuscitation in non-academic level II trauma centers

BackgroundEfficient resuscitation after trauma and shorter time to definitive hemorrhage control help improve trauma outcomes. We aimed to improve the speed and efficiency of resuscitation for critically ill trauma patients in the emergency department by involving interventional radiology and a second surgeon. Study designIn 2017 our community, non-academic level II trauma center implemented the Double 90 rule—for trauma patients with 2 confirmed systolic blood pressures <90 mm Hg—which involves a second activation including the interventional radiology team, backup trauma surgeon, and operating room charge nurse. We retrospectively reviewed our trauma registry to compare data for high-level trauma patients before (2016, “Pre-Dbl90”) and 3 consecutive years after intervention (2018–2020, “Dbl90”). ResultsAmong 613 patients who met criteria for our highest level of trauma activation, 100 either had activation of the Double 90 rule (Dbl90 patients, n = 76) or met Double 90 rule criteria (Pre-Dbl90 patients, n = 24). The groups were similar in age, sex, injury severity score, penetrating trauma incidence, and admission vitals. Median time to computed tomography decreased throughout the study period, from 34 min in 2016 to 18 min in 2020 (P < .001). Median time to first hemorrhage control procedure decreased from 118 min (2016) to 43 min (2020), (P = .013).Mean packed red blood cell transfusion decreased from 9.1 to 4.8 units (P = .016). Mortality rates were similar between groups. ConclusionThe Double 90 rule is effective for expediting trauma care starting in the emergency department, shortening the times to computed tomography, hemorrhage control intervention, and decreasing packed red blood cell transfusion.

A digital CRISPR-dCas9-based gene remodeling biocomputer programmed by dietary compounds in mammals

CRISPR-dCas9 (dead Cas9 protein) technology, combined with chemical molecules and light-triggered genetic switches, offers customizable control over gene perturbation. However, these simple ON/OFF switches cannot precisely determine the sophisticated perturbation process. Here, we developed a resveratrol and protocatechuic acid-programmed CRISPR-mediated gene remodeling biocomputer (REPACRISPR) for conditional endogenous transcriptional regulation of genes invitro and invivo. Two REPACRISPR variants, REPACRISPRi and REPACRISPRa, were designed for the logic control of gene inhibition and activation, respectively. We successfully demonstrated the digital computations of single or multiplexed endogenous gene transcription by using REPACRISPRa. We also established mathematical models to predict the dose-responsive transcriptional levels of a target endogenous gene controlled by REPACRISPRa. Moreover, high levels of endogenous gene activation in mice mediated by the AND logic gate demonstrated computational control of CRISPR-dCas9-based epigenome remodeling in mice. This CRISPR-based biocomputer expands the synthetic biology toolbox and can potentially advance gene-based precision medicine. A record of this paper's transparent peer review process is included in the supplemental information.

Understanding gait alterations: trunk flexion and its effects on walking neuromechanics.

Evolutionary and functional adaptations of morphology and postural tone of the spine and trunk are intrinsically shaped by the field of gravity in which humans move. Gravity also significantly impacts the timing and levels of neuromuscular activation, particularly in foot-support interactions. During step-to-step transitions, the centre of mass velocity must be redirected from downwards to upwards. When walking upright, this redirection is initiated by the trailing leg, propelling the body forward and upward before foot contact of the leading leg, defined as an anticipated transition. In this study, we investigated the neuromechanical adjustments when walking with a bent posture. Twenty adults walked on an instrumented treadmill at 4 km h-1 under normal (upright) conditions and with varying degrees of anterior trunk flexion (10, 20, 30 and 40deg). We recorded lower-limb kinematics, ground reaction forces under each foot, and the electromyography activity of five lower-limb muscles. Our findings indicate that with increasing trunk flexion, there is a lack of these anticipatory step-to-step transitions, and the leading limb performs the redirection after the ground collision. Surprisingly, attenuating distal extensor muscle activity at the end of stance is one of the main impacts of trunk flexion. Our observations may help us to understand the physiological mechanisms and biomechanical regulations underlying our tendency towards an upright posture, as well as possible motor control disturbances in some diseases associated with trunk orientation problems.

Relaxometric properties and biocompatibility of a novel nanostructured fluorinated gadolinium metal-organic framework.

A novel Gd-MOF based on tetrafluoro-terephthalic acid has been synthesized and its structure has been solved using X-ray single crystal diffraction data. The compound, with the formula [Gd2(F4BDC)3·H2O]·DMF, is isostructural with other Ln-MOFs based on the same ligand and has been recently reported. Its crystals were also reduced to nanometer size by employing acetic acid or cetyltrimethylammonium bromide (CTAB) as a modulator. The relaxometric properties of the nanoparticles were evaluated in solution by measuring 1H T1 and T2 as a function of the applied magnetic field and temperature. The biocompatibility of Gd-MOFs was evaluated on murine microglial BV-2 and human glioblastoma U251 cell lines. In both cell lines, Gd-MOFs do not modify the cell cycle profile or the activation levels of ERK1/2 and Akt, which are protein-serine/threonine kinases that participate in many signal transduction pathways. These pathways are fundamental in the regulation of a large variety of processes such as cell migration, cell cycle progression, differentiation, cell survival, metabolism, transcription, tumour progression and others. These data indicate that Gd-MOF nanoparticles exhibit high biocompatibility, making them potentially valuable for diagnostic and biomedical applications.

Detection of heat transfer mechanism in biological systems and HIF-1 α inducing invasion of liver cancer cells in hypoxic environments by activating STAT3

HIF-1α, as a hypoxia-inducing factor, can promote the invasion and metastasis of cancer cells by activating downstream signaling pathways. However, different heat transfer mechanisms in biological systems have important effects on the survival and function of tumor cells. The aim of this study was to investigate how the mechanisms of heat transfer in biological systems affect HIF-1α-mediated STAT3 activation and further induce the invasion ability of liver cancer cells in anoxic environment. The expression and activation of HIF-1α and STAT3 in hepatocellular carcinoma cell lines were detected by establishing anoxic model. At the same time, thermal imaging and heat transfer analysis methods were used to evaluate the heat transfer characteristics of cells under different temperature and hypoxia conditions. The effects of HIF-1α and STAT3 on invasion of hepatocellular carcinoma cells were analyzed in combination with cell migration and invasion experiments. The results showed that the expression of HIF-1α was significantly increased and the STAT3 signaling pathway was activated under hypoxia. The abnormal change of the heat transfer mechanism of biological system accelerates the invasion ability of hepatocellular carcinoma cells. Inhibition of HIF-1α expression can significantly reduce the activation level of STAT3, thereby inhibiting cell migration and invasion. This study reveals the interaction between the biological heat transfer mechanism and HIF-1α-STAT3 signaling pathway under hypoxia environment, which provides a new potential target for the treatment of malignant tumors such as liver cancer, and emphasizes the importance of regulating the heat transfer mechanism.

How does prolonged tennis playing affect lower limb muscles' activity during first and second tennis serves?

We examined the effect of prolonged tennis playing on lower limb muscles' activity during the execution of first and second tennis serves. Ten male competitive tennis players executed five first and second serves before (pretest) and after (posttest) a 3-h tennis match. Surface electromyographic (EMG) activity of four lower limb muscles (vastus lateralis, rectus femoris, gastrocnemius lateralis, and soleus muscles) on each leg was recorded along with maximum ball velocity measured by a radar gun and peak vertical forces recorded by a force platform. For the vastus lateralis, gastrocnemius lateralis, and soleus muscles of the left leg as well as the vastus lateralis muscle of the right leg, EMG amplitude decreased from pre- to posttests (p≤0.033). These reductions in the EMG signal were generally more pronounced in the first serve (i.e., ranging from -10% to -40%) compared to the second serve (0% to -25%). Maximum ball velocity for both first (159±12 vs. 154±12km/h) and second (126±20 vs. 125±15km/h) serves remained unchanged from pre- to posttests (p=0.638) Similarly, peak vertical forces did not differ between pretest and posttest for both first (1.78±0.30 vs. 1.72±0.29 body weight) and second (1.62±0.25 vs. 1.75±0.23 body weight) serves (p=0.730). In conclusion, a 3-h tennis match led to decreased activation levels in various leg muscles during serves, particularly in first serves compared to second serves. Despite consistent maximum ball velocity and peak vertical forces, these reductions in EMG signals suggest that skilled tennis players may adopt compensatory strategies after prolonged play.

A Triple-Mismatch Differentiating assay exploiting activation and trans cleavage of CRISPR-Cas12a for mutation detection with ultra specificity and sensitivity

Liquid biopsy technology is non-invasive and convenient, and is currently an emerging technology for cancer screening. Among them, clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein 12a (Cas12a) based nucleic acid detection technology has the advantages of high sensitivity, rapidity, and easy operation. However, CRISPR-Cas12a does not discriminate single-base mismatches of targets well enough to meet the needs of clinical detection. Herein, we developed the Triple-Mismatch Differentiating (TMD) assay. This assay amplified the small thermodynamic difference in mismatches at one site at the level of CRISPR-Cas12a activation to a significant thermodynamic difference at three sites at both the level of CRISPR-Cas12a activation and trans-cleavage, which greatly improves the ability of CRISPR-Cas12a to discriminate between base mismatches. Our manipulation greatly improved the specificity of the CRISPR-Cas12a system while maintaining its inherent sensitivity and simplicity, increasing the detection limit to 0.0001%. When testing samples from pancreatic cancer patients, our results were highly consistent with NGS sequencing results. We believe that the TMD assay will provide a new technology for early cancer detection and will be widely used in the clinical practice.

Mild therapeutic hypothermic protection activates the PI3K/AKT signaling pathway to inhibit TRPM7 and suppress ferroptosis induced by myocardial ischemia‑reperfusion injury.

The present study aimed to investigate the role of PI3K‑mediated ferroptosis signaling induced by mild therapeutic hypothermia (MTH), which was defined as a temperature of 34˚C, in protecting against myocardial ischemia-reperfusion (I/R) injury (MIRI). To meet this aim, H9C2 cells underwent hypoxia‑reperfusion (H/R) and/or MTH. The MTT assay was used to assess cell viability, cytotoxicity was measured using a lactate dehydrogenase cytotoxicity assay, and Annexin V‑FITC/PI flow cytometric analysis was used to analyze early and late cell apoptosis. In addition, 84 healthy adult male Sprague‑Dawley rats were randomly divided into seven groups (n=12), and underwent I/R and various treatments. Hemodynamics were monitored, and the levels of myocardial injury marker enzymes and oxidative stress markers in myocardial tissue were measured using ELISA. The expression levels of PI3K, AKT, transient receptor potential cation channel subfamily M member 7 (TRPM7), glutathione peroxidase 4 (GPX4) and acyl‑CoA synthetase long chain family member 4 (ACSL4) in animals and cells were measured using western blot analysis. These experiments revealed that MTH could effectively reduce myocardial infarct size, improve hemodynamic performance following MIRI and suppress myocardial apoptosis, thereby contributing to the recovery from H/R injury. Mechanistically, MTH was revealed to be able to activate the PI3K/AKT signaling pathway in cells, upregulating GPX4, and downregulating the expression levels of TRPM7 and ACSL4. Treatment with 2‑aminoethoxydiphenyl borate (an inhibitor of TRPM7) could further strengthen the myocardial protective effects of MTH, whereas treatment with erastin (promoter of ferroptosis) and wortmannin (inhibitor of PI3K) led to the effective elimination of the myocardial protective effects of MTH. Compared with in the I/R group, the PI3K/AKT activation level and the expression levels of GPX4 were both significantly increased, whereas the expression levels of TRPM7 and ACSL4 were significantly decreased in the I/R + MTH group. Taken together, the results of the present study indicated that MTH may activate the PI3K/AKT signaling pathway to inhibit TRPM7 and suppress ferroptosis induced by MIRI.