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Efferocytosis refers to the process by which phagocytes remove apoptotic cells and related apoptotic products. It is essential for the growth and development of the body, the repair of damaged or inflamed tissues, and the balance of the immune system. Damaged efferocytosis will cause a variety of chronic inflammation and immune system diseases. Many studies show that efferocytosis is a process mediated by mitochondria. Mitochondrial metabolism, mitochondrial dynamics, and communication between mitochondria and other organelles can all affect phagocytes' clearance of apoptotic cells. Therefore, targeting mitochondria to modulate phagocyte efferocytosis is an anticipated strategy to prevent and treat chronic inflammatory diseases and autoimmune diseases. In this review, we introduced the mechanism of efferocytosis and the pivoted role of mitochondria in efferocytosis. In addition, we focused on the therapeutic implication of drugs targeting mitochondria in diseases related to efferocytosis dysfunction. [ABSTRACT FROM AUTHOR]

Temperature as an environmental parameter influences the evolution of an open quantum system. In detail, temperature lies in Lindblad operator of quantum master equation that the evolution of an open quantum system follows. Hence, one can implement a temperature estimation of thermal baths through a measurement of quantum Fisher information about temperature brought from quantum states. Such a method by calculating quantum Fisher information about a parameter to estimate its value avoids measuring the parameter directly and it does not change the value of the parameter due to making measurements. In this paper, we consider a model consisting of a XXZ spin- 1 2 chain coupled locally to independent thermal baths with different temperature. Based on the model, we investigate optimal temperature estimation for thermal baths with respect to an open quantum system subjected to non-steady states. We first study optimal probe time for temperature estimation in the case of non-steady states and find that the optimal time shows different features for different types of system variables. It proves that in a certain duration there exists a tradeoff between the trial times and the attaining amount of Fisher information in each trial. In addition, we pay attention to an issue on optimal probe states. We demonstrate that in many cases the optimal states are not always the maximally entangled states and even maybe the separable states, which is related with the measuring time, system couplings. [ABSTRACT FROM AUTHOR]

We propose a scheme for generating squeezed states of a nanomechanical resonator. The scheme is based on a hybrid system consisting of two NAMRs and a superconducting charge qubit (SCCQ). The nonlinear interaction between the two nanomechanical resonators (NAMRs) can be implemented by setting the external biased flux of the SCCQ at some certain values, which plays the role of 'nonlinear media'. The noise in the SCCQ does not need to be considered, since we can adiabatically keep the SCCQ at the ground state. In addition, the squeezing parameters can also be adjusted by changing the external driving voltage. [ABSTRACT FROM AUTHOR]

When testing small size and high density objects with a broad field-of-view (FOV) industrial CT system, multiple objects are always assembled evenly onto the turntable for detecting to improve test efficiency. However, the maximum X-ray’s penetrating path through the materials increases, which means the CT system should collocate with a high energy X-ray source and wide-dynamic range detectors to complete the CT scanning. In this study, we proposed and tested a novel and efficient CT scanning method based on linear-arrangement and synchronous-rotating multi-turntables without enhancing the energy of X-ray source and wide-dynamic range of detectors for the CT system. With this modality, multiple objects are assembled onto multiple synchronous-rotating turntables respectively, and X-rays within the FOV merely penetrate one single object when scanning. The corresponding filtered back projection algorithm for image reconstruction is deduced. The computer simulation and experimental results verified the feasibility of this novel method and the scanning time was reduced to 5–8 minutes when completing the scanning of 3 to 5 group objects. [ABSTRACT FROM AUTHOR]

Diabetes is strongly associated with cognitive decline, but the molecular reasons are unknown. We found that fasting and peripheral insulin promote phosphorylation and dephosphorylation, respectively, of specific residues on brain proteins including cytoskeletal regulators such as slit-robo GTPase-activating protein 3 (srGAP3) and microtubule affinity-regulating protein kinases (MARKs), in which deficiency or dysregulation is linked to neurological disorders. Fasting activates protein kinase A (PKA) but not PKB/Akt signaling in the brain, and PKA can phosphorylate the purified srGAP3. The phosphorylation of srGAP3 and MARKs were increased when PKA signaling was activated in primary neurons. Knockdown of PKA decreased the phosphorylation of srGAP3. Furthermore, WAVE1, a protein kinase A-anchoring protein, formed a complex with srGAP3 and PKA in the brain of fasted mice to facilitate the phosphorylation of srGAP3 by PKA. Although brain cells have insulin receptors, our findings are inconsistent with the down-regulation of phosphorylation of target proteins being mediated by insulin signaling within the brain. Rather, our findings infer that systemic insulin, through a yet unknown mechanism, inhibits PKA or protein kinase(s) with similar specificity and/or activates an unknown phosphatase in the brain. Ser858 of srGAP3 was identified as a key regulatory residue in which phosphorylation by PKA enhanced the GAP activity of srGAP3 toward its substrate, Rac1, in cells, thereby inhibiting the action of this GTPase in cytoskeletal regulation. Our findings reveal novel mechanisms linking peripheral insulin sensitivity with cytoskeletal remodeling in neurons, which may help to explain the association of diabetes with neurological disorders such as Alzheimer disease. [ABSTRACT FROM AUTHOR]

UF-growth algorithm generated a large number of tree nodes and branches, and it needed to constantly calculate the support of candidate data items. To solve these problems, this paper proposed a new algorithm named compressed UF-tree. The algorithm changed the condition of constructing tree. Once the data item in transaction matched the tree node in a branch, merged the data item to the branch. Otherwise, it created a new branch from the tree node. Finally, it saved TID of transaction in the leaf node. Building probability vector of single data item and calculating the support of candidate data items through TID and probability vector to mine frequent items. It proves that it's feasible and efficient according to experimental comparison and analysis. [ABSTRACT FROM AUTHOR]

A library of isoxazole and 1,2,4-oxadiazole-containing diheterocyclic compounds has been prepared. Our strategy was explored in solution phase first as follows. PMB-protected 3-butyn-2-ol was deprotonated with nBuLi, acylated with methyl chloroformate, and then employed in a nitrile oxide 1,3-dipolar cycloaddition (benzaldehyde oxime in the presence of bleach) to afford the isoxazolesubstituted carboxylic acid methyl ester. Ester saponification with aqueous NaOH followed by a two-step condensation with benzamidoxime gave the final isoxazole-oxadiazole diheterocyclic product in good yield. With some modifications, we next explored this chemistry on Wang resin, which led to 18 final products that were cleaved from polymer beads with 50% TFA in dichloromethane. [ABSTRACT FROM AUTHOR]

We propose a scheme for implementing quantum information transfer based on frequency modes of microwave photons in a superconducting circuit. In our proposal, quantum information can be encoded on frequency modes of microwave photons, which act as a qubit in the resonator. Operations for the qubit, which is a process involving parametric frequency conversion, can be implemented by adjusting biased-dc superconducting quantum interference (SQUID). The coupling between two resonators can be controlled by tuning the frequency of the LC circuit inserted by a dc SQUID with two Josephson-junctions (2JJ-SQUID). Compared with previous ones, our work can avoid dephasing and decoherence resulting from atom decay. In addition, the resonator which includes multiple photons in two frequency modes can play a role of an identical atomic ensemble, which could lead to photon blockade [ABSTRACT FROM AUTHOR]

Integrated demand response (IDR) is deemed as an effective tool to balance energy supply and demand. User's uncertain information containing prior uncertain information and posterior uncertain information is a key factor affecting the implementation effectiveness of IDR, but existing studies fail to consider the two types of uncertain information, response risk caused by the uncertain information, and risk appetite comprehensively. Based on the principal‐agent theory of optimal incentive contract under uncertain information and Markowitz's mean‐variance portfolio theory, a new IDR model is established in this paper, and an IDR optimization strategy considering risk appetite under uncertain information is proposed. By proposing the user model considering multi‐dimensional uncertain information and the risk appetite‐based integrated energy service providers (IESP) model based on the principal‐agent theory and Markowitz's mean‐variance portfolio theory, we have achieved effective modelling of the user's uncertain information and the risk borne by IESP. The arithmetic examples have verified advantages of the model in enhancing the accuracy of user's actual response prediction and the superiority of incentive strategies, which is beneficial to reduce the cost of IESPs and enhance the benefit of users participating in IDR. [ABSTRACT FROM AUTHOR]

The essential role of intraflagellar transport protein IFT81 in male mice spermiogenesis and fertility. Am J Physiol Cell Physiol 318: C1092-C1106, 2020. First published April 1, 2020; doi:10.1152/ajpcell.00450.2019.--Intraflagellar transport (IFT) is an evolutionarily conserved mechanism that is indispensable for the formation and maintenance of cilia and flagella; however, the implications and functions of IFT81 remain unknown. In this study, we disrupted IFT81 expression in male germ cells starting from the spermatocyte stage. As a result, homozygous mutant males were completely infertile and displayed abnormal sperm parameters. In addition to oligozoospermia, spermatozoa presented dysmorphic and nonfunctional flagella. Histological examination of testes from homozygous mutant mice revealed abnormal spermiogenesis associated with sloughing of germ cells and the presence of numerous multinucleated giant germ cells (symblasts) in the lumen of seminiferous tubules and epididymis. Moreover, only few elongated spermatids and spermatozoa were seen in analyzed cross sections. Transmission electron microscopy showed a complete disorganization of the axoneme and para-axonemal structures such as the mitochondrial sheath, fibrous sheath, and outer dense fibers. In addition, numerous vesicles that contain unassembled microtubules were observed within developing spermatids. Acrosome structure analysis showed normal appearance, thus excluding a crucial role of IFT81 in acrosome biogenesis. These observations showed that IFT81 is an important member of the IFT process during spermatogenesis and that its absence is associated with abnormal flagellum formation leading to male infertility. The expression levels of several IFT components in testes, including IFT20, IFT25, IFT27, IFT57, IFT74, and IFT88, but not IFT140, were significantly reduced in homozygous mutant mice. Overall, our study demonstrates that IFT81 plays an essential role during spermatogenesis by modulating the assembly and elongation of the sperm flagella. [ABSTRACT FROM AUTHOR]

Cardiovascular disease is the leading cause of death worldwide, and it's of great importance to understand its underlying mechanisms and find new treatments. Sphingosine 1-phosphate (S1P) is an active lipid that exerts its effects through S1P receptors on the cell surface or intracellular signal, and regulates many cellular processes such as cell growth, cell proliferation, cell migration, cell survival, and so on. S1PR modulators are a class of modulators that can interact with S1PR subtypes to activate receptors or block their activity, exerting either agonist or functional antagonist effects. Many studies have shown that S1P plays a protective role in the cardiovascular system and regulates cardiac physiological functions mainly through interaction with cell surface S1P receptors (S1PRs). Therefore, S1PR modulators may play a therapeutic role in cardiovascular diseases. Here, we review five S1PRs and their functions and the progress of S1PR modulators. In addition, we focus on the effects of S1PR modulators on atherosclerosis, myocardial infarction, myocardial ischaemia/reperfusion injury, diabetic cardiovascular diseases, and myocarditis, which may provide valuable insights into potential therapeutic strategies for cardiovascular disease. [ABSTRACT FROM AUTHOR]

We propose a scheme for generating squeezed states based on a superconducting hybrid system. Our system consists of a nanomechanical resonator, a superconducting flux qubit, and a superconducting transmission line resonator. Using our proposal, one can easily generate the squeezed states of the nanomechanical resonator. In our scheme, the nonlinear interaction between the nanomechanical resonator and the superconducting transmission line resonator can be implemented by the flux qubit as ‘nonlinear media’ with a tunable Josephson energy. The realization of the nonlinearity does not need any operations on the flux qubit and just needs to adiabatically keep it at the ground state, which can greatly decrease the effect of the decoherence of the flux qubit on the squeezed efficiency. [ABSTRACT FROM AUTHOR]

[Objectives] To observe the effect of motor relearning combined with transcranial direct current stimulation on the motor function of lower extremities in patients with cerebral infarction, and to observe its effect on gait by 3D gait analysis. [Methods] 60 patients with cerebral infarction who met the inclusion criteria were randomly divided into 3 groups according to the order of treatment (n =20). Group A received motor relearning treatment, group B received transcranial direct current stimulation treatment, group C received motor relearning combined with transcranial direct current stimulation, and the curative effect was observed after 5 courses of treatment. [Results] Before treatment, FMA, MBI, spatio-temporal parameters for 3D gait analysis (gait frequency, gait cycle, stride length, gait speed, stride length deviation, double support) and lower limb joint motion parameters (affected side stride length, maximum hip flexion, maximum hip extension, maximum knee flexion, maximum knee extension, stance phase, swing phase) were compared among the three groups. After treatment, the FMA and MBI of the three groups increased, and the spatio-temporal parameters for 3D gait analysis (gait frequency, gait cycle, gait speed, double support) and the lower limb joint motion parameters (affected side stride length, maximum hip flexion, maximum hip extension, maximum knee flexion, swing phase) were all improved, while the spatio-temporal parameters (stride length and stride length deviation) and the lower limb joint motion parameters (maximum knee extension and stance phase) decreased. Compared with those before treatment, there were significant differences among the three groups (P < 0.05) . Through the comparison between groups, it was found that the FMA, MBI, spatio-temporal parameters for 3D gait analysis (gait frequency, gait cycle, gait speed, double support) and lower limb joint motion parameters (affected side stride length, maximum hip flexion, maximum hip extension, maximum knee flexion, swing phase) in group C were significantly higher than those in group A and B, while the spatio-temporal parameters (stride length and stride length deviation) and lower limb joint motion parameters (maximum knee extension and stance phase) in group C were significantly lower than those in group A and group B, and the difference was statistically significant (P <0.05). [ Conclusions] Motor relearning combined with transcranial direct current stimulation could in-crease MBI and FMA, improve gait spatio-temporal parameters and lower limb joint motion parameters, and correct abnormal gait in patients with cerebral infarction. [ABSTRACT FROM AUTHOR]

Gene transcription analysis is important in cancer research, and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) has been demonstrated to be an effective method to evaluate gene transcription in cancer. RT-qPCR requires an internal reference gene with a consistent level of mRNA transcription across various experimental conditions. However, it has been suggested that different treatments, including anticancer therapy, may influence the transcriptional stability of internal reference genes. Paclitaxel (PTX) and 10-hydroxycamptothecin (HCPT) are widely used to treat various types of cancer, and a suitable internal reference gene is required in order to analyze the transcription profiles of the cells following treatment. In the current study, the transcriptional stability of 30 candidate reference genes was investigated in cancer cells following treatment with PTX and HCPT. The two ovarian cancer cell lines, UACC-1598 and SKOV3, were treated with PTX and HCPT for 24 and 48 h, and the transcriptional levels of the candidate reference genes were subsequently evaluated by RT-qPCR analysis. The transcriptional stability of the selected genes was then analyzed using qbase+ and NormFinder software. A total of 9 genes were demonstrated to exhibit high transcriptional stability and one of these genes, ribosomal protein L13a (RPL13A), was identified to exhibit high transcriptional stability in every group. The current study identified various reference genes suitable under different circumstances, while RPL13A was indicated to be the most suitable reference gene for analyzing the transcription profile of ovarian cancer cells following treatment with PTX and HCPT. [ABSTRACT FROM AUTHOR]

AS160 (Akt substrate of 160 kDa) is a Rab GTPase-activating protein implicated in insulin control of GLUT4 (glucose transporter 4) trafficking. In humans, a truncation mutation (R363X) in one allele of AS160 decreased the expression of the protein and caused severe postprandial hyperinsulinaemia during puberty. To complement the limited studies possible in humans, we generated an AS160-knockout mouse. In wild-type mice, AS160 expression is relatively high in adipose tissue and soleus muscle, low in EDL (extensor digitorum longus) muscle and detectable in liver only after enrichment. Despite having lower blood glucose levels under both fasted and randomfed conditions, the AS160-knockout mice exhibited insulin resistance in both muscle and liver in a euglycaemic clamp study. Consistent with this paradoxical phenotype, basal glucose uptake was higher in AS160-knockout primary adipocytes and normal in isolated soleus muscle, but their insulin-stimulated glucose uptake and overall GLUT4 levels were markedly decreased. In contrast, insulin-stimulated glucose uptake and GLUT4 levels were normal in EDL muscle. The liver also contributes to the AS160-knockout phenotype via hepatic insulin resistance, elevated hepatic expression of phosphoenolpyruvate carboxykinase isoforms and pyruvate intolerance, which are indicative of increased gluconeogenesis. Overall, as well as its catalytic function, AS160 influences expression of other proteins, and its loss deregulates basal and insulin-regulated glucose homoeostasis, not only in tissues that normally express AS160, but also by influencing liver function. [ABSTRACT FROM AUTHOR]

Lantibiotics are ribosomally synthesized peptides that undergo posttranslational modifications to their mature, antimicrobial form. They are characterized by the unique amino acids lanthionine and methyllanthionine, introduced by means of dehydration of Ser/Thr residues followed by reaction of the resulting dehydro amino acids with cysteines to form thioether linkages. Two-component lantibiotics use two peptides that are each posttranslationally modified to yield two functionally distinct products that act in synergy to provide bactericidal activity. By using genetic data instead of isolation, a two-component lantibiotic, haloduracin, was identified in the genome of the Gram-positive alkaliphilic bacterium Bacillus halodurans C-125. We show that heterologously expressed and purified precursor peptides HalA1 and HalA2 are processed by the purified modification enzymes HalM1 and HalM2 in an in vitro reconstitution of the biosynthesis of a two-component lantibiotic. The activity of each HalM enzyme is substrate-specific, and the assay products exhibit antimicrobial activity after removal of their leader sequences at an engineered Factor Xa cleavage site, indicating that correct thioether formation has occurred. Haloduracin's biological activity depends on the presence of both modified peptides. The structures of the two mature haloduracin peptides Halα and Halβ were investigated, indicating that they have similarities as well as some distinct differences compared with other two-component lantibiotics. [ABSTRACT FROM AUTHOR]

Systemic lupus erythematosus (SLE) is an autoimmune disease posing threats to multiple organs in the human body. As a typical manifestation of SLE, lupus nephritis is characterized by a series of pathological changes in glomerulus as well as accumulation of pathogenic autoreactive IgG with complement in the kidney that dramatically disrupts renal functions. Activation-induced deaminase (AID), which governs both somatic hypermutation (SHM) and class-switch recombination (CSR), has been shown to be essential for the regulation of SLE. However, the relative contributions of SHM and CSR to SLE pathology have not been determined. Based on the available AIDG23S mice, we successfully established an AIDG23S MRL/lpr mouse model, in which SHM is specifically abolished, although CSR is largely unaffected. We found that the abrogation of SHM effectively alleviated SLE-associated histopathological alterations, such as expansion of the mesangial matrix and thickening of the basement membrane of Bowman's capsule as well as infiltration of inflammatory cells. Compared with SLE mice, AIDG23S MRL/lpr mice exhibited decreased proteinuria, blood urea nitrogen, and creatinine, indicating that the loss of SHM contributed to the recovery of renal functions. As a consequence, the life span of those SHM-deficient MRL/lpr mice was extended. Together, we provide direct evidence pinpointing a vital role of SHM in the control of SLE development. [ABSTRACT FROM AUTHOR]

Skeletal muscle absorbs long-chain fatty acids (LCFAs) that are either oxidized in mitochondria or temporarily stored as triglycerides in lipid droplets (LDs). So far, it is still not fully understood how lipid uptake and storage are regulated in muscle and whether these are important for whole-body lipid homeostasis. Here we show that the small GTPase Rab8a regulates lipid uptake and storage in skeletal muscle. Muscle-specific Rab8a deletion caused hyperlipidemia and exacerbated hepatosteatosis induced by a high-fat diet. Mechanistically, Rab8a deficiency decreased LCFA entry into skeletal muscle and inhibited LD fusion in muscle cells. Consequently, blood lipid levels were elevated and stimulated hepatic mammalian target of rapamycin, which enhanced hepatosteatosis by upregulating hepatic lipogenesis and cholesterol biosynthesis. Our results demonstrate the significance of lipid uptake and storage in muscle in regulating whole-body lipid homeostasis, and they shed light on the roles of skeletal muscle in the pathogenesis of hyperlipidemia and hepatosteatosis. [ABSTRACT FROM AUTHOR]

Tre-2/USP6, BUB2, cdc16 domain family member 1 (the TBC domain is the GTPase activating protein domain) (TBC1D1) is a Rab GTPase activating protein that is phosphorylated on Ser231 by the AMP-activated protein kinase (AMPK) in response to intracellular energy stress. However, the in vivo role and importance of this phosphorylation event remains unknown. To address this question, we generated a mouse model harboring a TBC1D1Ser231Ala knockin (KI) mutation and found that the KI mice developed obesity on a normal chow diet. Mechanistically, TBC1D1 is located on insulin-like growth factor 1 (IGF1) storage vesicles, and the KI mutation increases endocrinal and paracrinal/autocrinal IGF1 secretion in an Rab8a-dependent manner. Hypersecretion of IGF1 causes increased expression of lipogenic genes via activating the protein kinase B (PKB; also known as Akt)–mammalian target of rapamycin (mTOR) pathway in adipose tissues, which contributes to the development of obesity, diabetes, and hepatic steatosis as the KI mice age. Collectively, these findings demonstrate that the AMPK–TBC1D1 signaling nexus interacts with the PKB–mTOR pathway via IGF1 secretion, which consequently controls expression of lipogenic genes in the adipose tissue. These findings also have implications for drug discovery to combat obesity. [ABSTRACT FROM AUTHOR]

[Display omitted] • •Free-standing V 2 O 3 /VN/C interlayers are constructed to suppress shuttle effect. • •The V 2 O 3 /VN catalysts with adsorption and catalysis enhance redox kinetics. • •The cells with V 2 O 3 /VN/C interlayers exhibit high capacity and long cycle stability. The shuttle effect caused by soluble lithium polysulfides (LiPSs) hinders the practical application of lithium–sulfur batteries. The sluggish reaction kinetics of LiPSs results in rapid capacity decay. In this work, we constructed a free-standing multifunctional interlayer decorated with V 2 O 3 /VN catalysts to suppress the shuttle effect and boost the redox reaction kinetics. The introduction of VN with strong chemical adsorption improves the conductivity of V 2 O 3 , which is beneficial for the kinetics of conversion reaction. In addition, the micro/mesopores inside the structure not only facilitate ion diffusion but also buffer the volume change of the sulfur cathode during the cycling process. Benefiting from the synergistic effects, the fabricated cells exhibit a high specific capacity of 1419.5 mAh g−1 at 0.1C. The cell with V 2 O 3 /VN/C exhibits a promising cycling performance over 1000 cycles with a decay rate of 0.037% per cycle at 1C. The synergistic effects of electrocatalysts and conductive carbon material provide inspiration on the development of high-performance Li–S batteries. [ABSTRACT FROM AUTHOR]

Single-base substitution may affect the function of genes. This study identified a single-base substitution of G for A in codon 148 of cyclin-dependent kinase inhibitor 2A (CDKN2A/p16) by sequencing human ovarian cancer cell line UACC-1598. As a tumor suppressor gene, the expression of CDKN2A/p16 should be strictly controlled. In order to control CDKN2A/p16 gene expression, an inducible pTUNE vector system was selected. Using recombinant DNA technology, a CDKN2A/p16-A148T and CDKN2A/p16-wi ld-type gene expression system was successfully constructed to investigate whether this single-base substitution affects the function of CDKN2A/p16. For the wild-type and the mutant, expression of CDKN2A/p16-green fluorescent protein fusion protein increased markedly following isopropyl-β-D-thiogalactoside induction, and was accompanied by signiicant G1 arrest in the transfected human ovarian cancer SKOV3 cell line. The inducible vectors used in this study, CDKN2A/p16-wild-type and CDKN2A/p16-A148T open reading frame, may be useful for further investigation into whether this somatic mutation could alter the function of CDKN2A/p16 as a tumor suppressor gene. In summary, CDKN2A/p16-A148T was identified in ovarian cancer cells, and this single-base substitution did not affect the ability of CDKN2A/p16 to arrest the cell cycle. [ABSTRACT FROM AUTHOR]

Background: Rho GTPases are involved in cellular functions relevant to cancer. The roles of RhoA and Rac1 have already been established. However, the role of Rac3 in cancer aggressiveness is less well understood. Methods: This work was conducted to analyze the implication of Rac3 in the aggressiveness of two breast cancer cell lines, MDA-MB-231 and MCF-7: both express Rac3, but MDA-MB-231 expresses more activated RhoA. The effect of Rac3 in cancer cells was also compared with its effect on the non-tumorigenic mammary epithelial cells MCF- 10A. We analyzed the consequences of Rac3 depletion by anti-Rac3 siRNA. Results: Firstly, we analyzed the effects of Rac3 depletion on the breast cancer cells' aggressiveness. In the invasive MDA-MB-231 cells, Rac3 inhibition caused a marked reduction of both invasion (40%) and cell adhesion to collagen (84%), accompanied by an increase in TNF-induced apoptosis (72%). This indicates that Rac3 is involved in the cancer cells' aggressiveness. Secondly, we investigated the effects of Rac3 inhibition on the expression and activation of related signaling molecules, including NF-κB and ERK. Cytokine secretion profiles were also analyzed. In the non-invasive MCF-7 line; Rac3 did not influence any of the parameters of aggressiveness. Conclusions: This discrepancy between the effects of Rac3 knockdown in the two cell lines could be explained as follows: in the MDA-MB-231 line, the Rac3-dependent aggressiveness of the cancer cells is due to the Rac3/ERK-2/ NF-κB signaling pathway, which is responsible for MMP-9, interleukin-6, -8 and GRO secretion, as well as the resistance to TNF-induced apoptosis, whereas in the MCF-7 line, this pathway is not functional because of the low expression of NF-κB subunits in these cells. Rac3 may be a potent target for inhibiting aggressive breast cancer. [ABSTRACT FROM AUTHOR]

1. Metformin is an activator of AMP-activated protein kinase (AMPK). Recent studies suggest that pharmacological activation of AMPK inhibits cardiac hypertrophy. In the present study, we examined whether long-term treatment with metformin could attenuate ventricular hypertrophy in a rat model. The potential involvement of nitric oxide (NO) in the effects of metformin was also investigated. 2. Ventricular hypertrophy was established in rats by transaortic constriction (TAC). Starting 1 week after the TAC procedure, rats were treated with metformin (300 mg/kg per day, p.o.), N-nitro-arginine methyl ester (-NAME; 50 mg/kg per day, p.o.) or both for 8 weeks prior to the assessment of haemodynamic function and cardiac hypertrophy. 3. Cultured cardiomyocytes were used to examine the effects of metformin on the AMPK-endothelial NO synthase (eNOS) pathway. Cells were exposed to angiotensin (Ang) II (10 mol/L) for 24 h under serum-free conditions in the presence or absence of metformin (10 mol/L), compound C (10 mol/L), -NAME (10 mol/L) or their combination. The rate of incorporation of [H]-leucine was determined, western blotting analyses of AMPK-eNOS, neuronal nitric oxide synthase (nNOS) and inducible nitric oxide synthase (iNOS) were undertaken and the concentration of NO in culture media was determined. 4. Transaortic constriction resulted in significant haemodynamic dysfunction and ventricular hypertrophy. Myocardial fibrosis was also evident. Treatment with metformin improved haemodynamic function and significantly attenuated ventricular hypertrophy. Most of the effects of metformin were abolished by concomitant -NAME treatment. -NAME on its own had no effect on haemodynamic function and ventricular hypertrophy in TAC rats. 5. In cardiomyocytes, metformin inhibited AngII-induced protein synthesis, an effect that was suppressed by the AMPK inhibitor compound C or the eNOS inhibitor -NAME. The improvement in cardiac structure and function following metformin treatment was associated with enhanced phosphorylation of AMPK and eNOS and increased NO production. 6. The findings of the present study indicate that long-term treatment with metformin could attenuate ventricular hypertrophy induced by pressure overload via activation of AMPK and a downstream signalling pathway involving eNOS-NO. [ABSTRACT FROM AUTHOR]

HYPB isa human histone H3 lysine 36 (H3K36)-specific methyltransferase and acts as the ortholog of yeast Set2. This study explored the physiological function of mammalian HYPB using knockout mice. Homozygous disruption of Hypb impaired H3K36 trimethylation but not monoor dimethylation, and resulted in embryonic lethality at E10.5-E11.5. Severe vascular defects were observed in the Hypb-/- embryo, yolk sac, and placenta. The abnormally dilated capillaries in mutant embryos and yolk sacs could not be remodeled into large blood vessels or intricate networks, and the aberrantly rounded mesodermal cells exhibited weakened interaction with endothelial cells. The embryonic vessels failed to invade the labyrinthine layer of placenta, which impaired the embryonic-maternal vascular connection. These defects could not be rescued by wildtype tetraploid blastocysts, excluding the possibility that they were caused by the extraembryonic tissues. Consistent with these phenotypes, gene expression profiling in wild-type and Hypb yolk sacs revealed that the Hypb disruption altered the expression of some genes involved in vascular remodeling. At the cellular level, Hypb-/- embryonic stem cell-derived embryonic bodies, as well as in vitro-cultured human endothelial cells with siRNA-mediated suppression of HYPB, showed obvious defects in cell migration and invasion during vessel formation, suggesting an intrinsic role of Hypb in vascular development. Taken together, these results indicate that Hypb is required for embryonic vascular remodeling and provide a tool to study the function of H3K36 methylation in vasculogenesis/angiogenesis. [ABSTRACT FROM AUTHOR]

Background: It has been demonstrated that hypertrophic scar fibroblasts (HSFs) overexpress collagen messenger ribonucleic acid (mRNA) and protein, especially α1 collagen. Antisense nucleic acids are effective in inhibiting harmful or uncontrolled gene expression, suggesting that antisense ribonucleic acid (RNA) can effectively downregulate the expression of α1 collagen gene and attenuate the scars. Aims: This study was conducted to observe the effect of recombinant plasmid pREP9-COL1 on α1 collagen expression in HSFs and clarify the prospect of antisense RNA on scar treatment. Methods: The α1 collagen gene fragment including the region of 5' UTR to exon (229 bp) was cloned in the eukaryotic expression plasmid pREP9 in the antisense orientation relative to the RSV-LTR promoter to reconstruct the pREP9- COL1 plasmid. Then it was transferred into HSFs through lipofectamine. The expression of α1 collagen was examined by immunostaining, reverse-transcriptase polymerase chain reaction, and Western blots. Results: The recombinant plasmid pREP9-COL1 with a correct sequence was constructed successfully; pREP9-COL1 consistently inhibited human α1 collagen gene expression at both mRNA and protein levels. Conclusions: Antisense RNA was effective in downregulating α1 collagen expression of HSFs. Therefore, this approach offered a prospect of scar treatment by attenuation of α1 collagen production with antisense RNA. [ABSTRACT FROM AUTHOR]

Background: Rho GTPases are involved in cellular functions relevant to cancer. The roles of RhoA and Rac1 have already been established. However, the role of Rac3 in cancer aggressiveness is less well understood.Methods: This work was conducted to analyze the implication of Rac3 in the aggressiveness of two breast cancer cell lines, MDA-MB-231 and MCF-7: both express Rac3, but MDA-MB-231 expresses more activated RhoA. The effect of Rac3 in cancer cells was also compared with its effect on the non-tumorigenic mammary epithelial cells MCF-10A. We analyzed the consequences of Rac3 depletion by anti-Rac3 siRNA.Results: Firstly, we analyzed the effects of Rac3 depletion on the breast cancer cells' aggressiveness. In the invasive MDA-MB-231 cells, Rac3 inhibition caused a marked reduction of both invasion (40%) and cell adhesion to collagen (84%), accompanied by an increase in TNF-induced apoptosis (72%). This indicates that Rac3 is involved in the cancer cells' aggressiveness. Secondly, we investigated the effects of Rac3 inhibition on the expression and activation of related signaling molecules, including NF-κB and ERK. Cytokine secretion profiles were also analyzed. In the non-invasive MCF-7 line; Rac3 did not influence any of the parameters of aggressiveness.Conclusions: This discrepancy between the effects of Rac3 knockdown in the two cell lines could be explained as follows: in the MDA-MB-231 line, the Rac3-dependent aggressiveness of the cancer cells is due to the Rac3/ERK-2/NF-κB signaling pathway, which is responsible for MMP-9, interleukin-6, -8 and GRO secretion, as well as the resistance to TNF-induced apoptosis, whereas in the MCF-7 line, this pathway is not functional because of the low expression of NF-κB subunits in these cells. Rac3 may be a potent target for inhibiting aggressive breast cancer. [ABSTRACT FROM AUTHOR]