Enhancing Factor Research Articles

Critical role of Myc activation in mouse hepatocarcinogenesis induced by the activation of AKT and RAS pathways

MYC activation at modest levels has been frequently found in hepatocellular carcinoma. However, its significance in hepatocarcinogenesis has remained obscure. Here we examined the role of Myc activation in mouse liver tumours induced by hepatocytic expression of myristoylated AKT (AKT) and/or mutant HRASV12 (HRAS) via transposon-mediated gene integration. AKT or HRAS alone required 5 months to induce liver tumours, whereas their combination generated hepatocellular carcinoma within 8 weeks. Co-introduction of AKT and HRAS induced lipid-laden preneoplastic cells that grew into nodules composed of tumour cells with or without intracytoplasmic lipid, with the latter being more proliferative and associated with spontaneous Myc expression. AKT/HRAS-induced tumorigenesis was almost completely abolished when MadMyc, a competitive Myc inhibitor, was expressed simultaneously. The Tet-On induction of MadMyc in preneoplastic cells significantly inhibited the progression of AKT/HRAS-induced tumours; its induction in transformed cells suppressed their proliferative activity with alterations in lipid metabolism and protein translation. Transposon-mediated Myc overexpression facilitated tumorigenesis by AKT or HRAS, and when it was co-introduced with AKT and HRAS, diffusely infiltrating tumours without lipid accumulation developed as early as 2 weeks. Examination of the dose-responses of Myc in the enhancement of AKT/HRAS-induced tumorigenesis revealed that a reduction to one-third retained enhancing effect but three-times greater introduction damped the process with increased apoptosis. Myc overexpression suppressed the mRNA expression of proteins involved in the synthesis of fatty acids, and when combined with HRAS introduction, it also suppressed the mRNA expression of proteins involved in their degradation. Finally, the MYC-positive human hepatocellular carcinoma was characterized by the cytoplasm devoid of lipid accumulation, prominent nucleoli and a higher proliferative activity. Our results demonstrate that in hepatocarcinogenesis induced by both activated AKT and HRAS, activation of endogenous Myc is an enhancing factor and adequate levels of Myc deregulation further facilitate the process with alterations in cellular metabolism.

Regulation of PfEMP1-VAR2CSA translation by a Plasmodium translation-enhancing factor.

Pregnancy-associated malaria commonly involves the binding of Plasmodium falciparum-infected erythrocytes to placental chondroitin sulfate A (CSA) through the PfEMP1-VAR2CSA protein. VAR2CSA is translationally repressed by an upstream open reading frame. In this study, we report that the P. falciparum translation enhancing factor (PTEF) relieves upstream open reading frame repression and thereby facilitates VAR2CSA translation. VAR2CSA protein levels in var2csa-transcribing parasites are dependent on the expression level of PTEF, and the alleviation of upstream open reading frame repression requires the proteolytic processing of PTEF by PfCalpain. Cleavage generates a C-terminal domain that contains a sterile-alpha-motif-like domain. The C-terminal domain is permissive to cytoplasmic shuttling and interacts with ribosomes to facilitate translational derepression of the var2csa coding sequence. It also enhances translation in a heterologous translation system and thus represents the first non-canonical translation enhancing factor to be found in a protozoan. Our results implicate PTEF in regulating placental CSA binding of infected erythrocytes.

Characterization and hydrogen sorption behaviors of FeNiCr-carbon composites derived from Fe, Ni and Cr-containing polyacrylonitrile fibers prepared by electrospinning method

In order to enhance hydrogen storage capacity of carbonaceous materials through metal modification, FeNiCr-carbon composites were prepared by calcination of Fe, Ni and Cr-containing polyacrylonitrile (PAN) fibers fabricated by electrospinning method. Fe (III), Ni (II) and Cr (III) acetylacetonates (M (acac)n) were selected as metal sources. Increase of specific surface area and formation of micropores were observed on heat decomposition of M (acac)n particularly through introduction of steam. Maximal hydrogen content, 1.62 mass%, was obtained at 77 K under 0.8 MPa of hydrogen for a FeNiCr-carbon composite, which contained about 15.5 mass% of metals and had specific surface area of 501 m2 g−1. The hydrogen content exceeded the hydrogen physisorption limit, 2.34 mass% per 1000 m2 g−1, which was calculated on the basis of the commensurate–incommensurate transition with an enhancing factor ρ of 1.126. After hydrogenation at 653 K, no hydrogen desorption peaks were observed for FeNiCr powders derived from M (acac)n, and one peak at 828 K for a carbonaceous sample prepared from unmodified PAN fibers. From the most promising FeNiCr-carbon composite, another peak was recorded at 752 K in addition to the peak at 828 K. The former would be originated from hydrogen on novel sorption sites additionally created on the composite formation.

Evaluation of Antiobesity activity of Ethanolic extract of Cashew Apple against high fat diet induced obesity in rodents

Obesity has doubled since 1980, as per WHO report affecting more than 1900 million adults throughout the world. In recent studies, diets containing antioxidants and branch chain amino acids were demonstrated to have potential effects on fat utilization. Cashew apple juice comprising nutritional components including vitamin C and BCAS reported as an enhancing factor in fat oxidation during exercise and may improve endurance performance (Piyapong Prasertsri et al., 2013). Hence, the current work was planned with an objective to evaluate the anti‐obesity activity of the ethanol extract of cashew apple (ECA). To identify the active constituents, ECA was subjected for analysis using GC‐MS technique. High fat diet induced rodent model was used to induce obesity in the experimental animals. Adult male albino rats were randomized in to five groups (n=6) among which first group was kept on standard rodent chow (normal). The remaining four groups were assigned as obese control, standard & ECA treated groups which received high fat diet along with vehicle, atorvastatin (10 mg/kg, p.o) and ECA (200 & 400 mg/kg, p.o) respectively for 60 days. At the end of study, obese control group animals showed a significant increase in BMI & body wt, serum total cholesterol, triglycerides, LDL‐cholesterol and VLDL‐cholesterol compared to normal group. Administration of ECA (200mg/kg, 400mg/kg) exhibited significant decrease in serum total cholesterol, triglycerides, LDL‐cholesterol and VLDL‐cholesterol levels in ECA treated groups. In addition HDL‐cholesterol levels were also increased distinctly compared to the obese control groups. A notable elevation in the levels of serum creatinine, transaminases & alkaline phosphatase were observed with an intake of high fat diet. These effects were reversed by oral administration of ECA. A significant inhibition of serum malondialdehyde (MDA) and improvement in hepatic enzymatic (catalase, superoxide dismutase), non‐enzymatic(reduced glutathione) levels was also identified as an outcome after treatment with ECA (200mg/kg, 400mg/kg) and standarad (atorvastatin) compared to the obese control groups. Eventually, consumption of cashew apple may even reverse the fatty liver which was quite evident from histology images. GC‐MS analysis revealed the presence of 5‐Hydroxy‐2‐(hydroxymethyl)‐4H‐pyran‐4‐one; 2,3‐dihydro‐3,5‐dihydroxy‐6‐methyl‐4H‐pyran‐4‐one; Hexadecanoic acid; octadecanoic acid; 5‐(Hydroxymethyl)‐2‐furancarboxaldehyde; Furfural; 4‐H‐pyran‐4‐one,3,5, dihydro‐2‐methyl decanoic acid ethyl ester.Based up on the available experimental evidence it can be concluded that ethanolic cashew apple extract possess anti‐obesity activity and reinforce recommending consumption of cashew apple to modulate obesity.Support or Funding Information1. Sri Padmavati Mahila Visvavidyalayam, Tirupati, Andhra Pradesh, India.2. DST, Government of India.

Characterization of a molt-related myostatin gene (FmMstn) from the banana shrimp Fenneropenaeus merguiensis

Myostatin is an important member of the transforming growth factor (TGF) family that functions to regulate muscle growth in animals. In this study, the myostatin gene (FmMstn) and two slightly different (short and long forms) cDNAs of the banana shrimp Fenneropenaeus merguiensis were cloned and characterized. Similar to Mstn gene of the scallop, fish and mammal, FmMstn gene consists of 3 exons and 2 introns. The 2kb upstream promoter region of the FmMstn gene consists of putative response elements for myocyte enhancing factor (MEF2) and E-box factors. The longest open reading frame of the short Mstn consists of 1260bp encoding for a protein with 420 amino acid residues. The long FmMstn is almost identical to the short FmMstn with the exception of 8 amino acid insertions. FmMstn is most similar to the Mstn of Litopenaeus vannamei and Penaeus monodon sharing >92–98% amino acid sequence identity. Multiple sequence alignment results revealed high degree of amino acid conservation of the cysteine residues and mature peptide of the FmMstn with Mstn from other animals. FmMstn transcript was detected in the heart, muscle, optic nerve and thoracic ganglion. FmMstn transcript level in muscle is higher in early postmolt, decreases in intermolt and increases again towards ecdysis. Higher expression level of FmMstn is also observed in smaller shrimp of the same age. Knock-down of FmMstn gene by RNAi can cause a significant increase in molt cycle duration and failure of some shrimp to undergo ecdysis. Direct DNA sequencing results revealed that FmMstn gene is highly polymorphic and several potential SNPs have been identified. Some SNPs are associated with the size difference of the shrimp. In summary, the result of this study indicates that shrimp FmMstn gene is molt/growth-related and the presence of SNP suggests that it could be a candidate gene for shrimp genetic improvement research.

Self-Association and Conformational Stability of NAMPT Protein

Nicotinamide phosphoribosyltransferase (NAMPT), also known as Visfatin and Pre B-cell colony enhancing factor (PBEF), is a rate-limiting enzyme in the salvage pathway required for nicotinamide adenine dinucleotide (NAD) biosynthesis. It is a highly conserved 52-kDa protein, found in living species from bacteria to humans [1]. The protein serves as a cytokine and is involved in cellular regulation influencing cancer, ischemia, obesity, and type-II diabetes [2, 3]. Despite being involved in many regulatory pathways, there is a paucity of information concerning the function of NAMPT, due to the limitations of in vivo assays, and lack of expression systems for the protein. Here, we successfully expressed the NAMPT protein using the pET-SUMO expression vector in E.coli strain SHuffle containing a hexa-His tag for protein purification. Activity assays demonstrated functionality of the protein. Moreover, initial biophysical characterization of the protein using circular dichroism revealed secondary structural elements consistent with crystallographic data. Dynamic light scattering showed the protein exists as large oligomeric units potentially involved in the NAMPT signal amplification pathway. Hydropathy analysis indicated possible hydrophobic patches on the protein surface that explains the native oligomeric state. Most striking, we discovered that NAMPT can be solubilized in n-dodecyl-β-D-maltopyranoside detergent in monomeric form. These findings open opportunities for further structural and functional investigations. Presently we are optimizing conditions for NMR experiments on NAMPT protein. These methods [4] are complementary to X-ray crystallography, and provide valuable information on the structure and dynamics, offering an important tool for understanding biological functioning.[1] T. Wang et al. (2006) Nat. Struct. Mol. Biol 13, 661.[2] S. M. Camp et al. (2015) Sci. Rep. 5, 13135.[3] A. Fukuhara et al. (2005) Science 307, 426.[4] T. R. Molugu et al. (2016) Chem. Rev. (in press).

Increased amygdala reactivity following early life stress: a potential resilience enhancer role

BackgroundAmygdala hyper-reactivity is sometimes assumed to be a vulnerability factor that predates depression; however, in healthy people, who experience early life stress but do not become depressed, it may represent a resilience mechanism. We aimed to test these hypothesis examining whether increased amygdala activity in association with a history of early life stress (ELS) was negatively or positively associated with depressive symptoms and impact of negative life event stress in never-depressed adults.MethodsTwenty-four healthy participants completed an individually tailored negative mood induction task during functional magnetic resonance imaging (fMRI) assessment along with evaluation of ELS.ResultsMood change and amygdala reactivity were increased in never-depressed participants who reported ELS compared to participants who reported no ELS. Yet, increased amygdala reactivity lowered effects of ELS on depressive symptoms and negative life events stress. Amygdala reactivity also had positive functional connectivity with the bilateral DLPFC, motor cortex and striatum in people with ELS during sad memory recall.ConclusionsIncreased amygdala activity in those with ELS was associated with decreased symptoms and increased neural features, consistent with emotion regulation, suggesting that preservation of robust amygdala reactions may reflect a stress buffering or resilience enhancing factor against depression and negative stressful events.

Tcf7l2 Gene Polymorphism in T2dm with Patients in Turkish Population

Diabetes Mellitus (DM) is a group of metabolic diseases characterized by hyperglycemia resulting from defects of insulin secretion and/or increased cellular resistance to insulin. Chronic hyperglycemia and other metabolic disturbances of DM lead to long-term tissue and organ damage as well as dysfunction involving the eyes, kidneys, and nervous and vascular systems. Type 2 Diabetes mellitus; is the most common form of DM worldwide, and its prevalence is increasing. Its underlying defects can vary from predominant insulin resistance with relative insulin deficiency to a predominant insulin secretory defect with insulin resistance. Transcription 7-like 2 (TCF7L2) protein has been implicated in blood glucose homeostasis. The transcription factor 7-like 2 (TCF7L2) gene is a member of the T-cell factor (TCF)/lymphoid enhancing factor family which encodes a high mobility group (HMG) box-containing transcription factor that plays a key role in the Wnt signaling pathway. The key effector of the canonical Wnt signaling pathway (defined as Wnt pathway hereafter) is the bipartite transcription factor β-cat (β-catenin)/TCF, formed by β-cat and a member of the TCF family [TCF-1/TCF7, LEF-1, TCF-3/TCF7L1 and TCF-4/TCF7L2]. In the absence of Wnt signaling, these HMG box TCF proteins function in the nucleus as transcriptional repressors of the Wnt target genes. TCF7L2 gene is located in chromosome 10q25.3, spans 215.9 kb and contains 14 exons (NCBI build 36.2). However, a previous study has shown that TCF7L2 has 17 exons, of which five are alternative splicing. Genetic variants of gene are associated with increased risk of type 2 diabetes. Common polymorphisms of the transcription factor 7-like 2 gene (TCF7L2) have recently been associated with Type 2 Diabetes all population. The prevalence of Type 2 Diabetes shows wide ethnic and geographic variations. In this study, we analyzed three polymorphisms in the TCF7L2 gene, rs12255372, rs11196205 and rs7901695 using a case-control design in 163 individuals (113 unrelated T2DM patients and 50 normoglycemic controls) to assess their association with T2DM risk in the Turkish population.

Study of Explosive Power as Performance Enhancing Factor Among Kabaddi Players

The study of power as a performance-enhancing factor among Kabaddi players is essential to comprehend the relationship between power development and athletic performance in this dynamic sport. Power plays a crucial role in Kabaddi, as it influences the ability to generate force rapidly, efficiently, and explosively, enabling athletes to execute actions such as sprinting, jumping, and tackling opponents effectively. This abstract provides an overview of the methodologies used to investigate power in Kabaddi performance. Performance tests, including vertical jump tests, standing broad jump tests, and agility tests, assess power-related abilities. Strength training interventions, targeting power development, involve exercises such as squats, lunges, power cleans, and plyometrics. Biomechanical analysis using motion capture systems and force plates evaluates the mechanics of Kabaddi movements. Comparative studies compare power-related characteristics and performance outcomes between elite and non-elite players. Longitudinal studies track the progress of players over time, measuring changes in strength, speed, agility, and on-field performance. These research approaches aim to enhance our understanding of power's impact on Kabaddi performance, provide evidence-based recommendations for training, and contribute to the development of Kabaddi players.

Pre-B-cell colony enhancing factor expression and its clinical significance in colorectal cancer

Objective To explore the expression of pre-B-cell colony enhancing factor (PBEF) in colorectal cancer, and analyze the relationship between PBEF expression and the clinical pathological characte-ristics and its diagnostic and prognostic value. Methods Sixty-eight human colorectal tumor and normal colo-rectal tissues were collected, and the expression of PBEF was detected by immunohistochemical method. The relationship between the expression level of PBEF and the characteristics of tumors and prognosis was analyzed. Results The positive expression rate of PBEF in colorectal cancer tissues was 80.9%, higher than that in normal colorectal tissues, which was 29.4% (χ2=36.41, P 0.05), sex (χ2=0.39, P>0.05), histology type (χ2=0.54, P>0.05) or penetrating serosa (χ2=1.55, P>0.05), but significantly associated with the degree of differentiation (χ2=6.29, P 0.05). Conclusion The abnormal high expression of PBEF may be closely related to the occurrence and development of colorectal cancer. Key words: Colorectal neoplasms; Immunohistochemistry; Pre-B-cell colony enhancing factor

Mechanism of Tissue Factor (TF) Enhancing Factor (F)VIII Activity on FXa Generation in Initial Phase of Coagulation and Interaction Between TF and FVIII C2 Domain

In the cell-based coagulation model, factor (F)VIIa complex with tissue factor (TF) initiates the blood coagulation by generating FXa as the extrinsic tenase complex and activates FIX which composes the intrinsic tenase complex. We demonstrated that FVIIa/TF directly activated FVIII in an early coagulation phase (Soeda, JTH, 2010), and TF enhanced the intrinsic tenase activity via possible interaction with FVIIIa (Ogiwara, ASH, 2010). In this study, we clarified the enhancing mechanism of intrinsic tenase activity in the TF-related up-regulation of FVIII, and identified the TF-interactive region on FVIII.To explore the enhancing mechanism of TF for FVIII regulation, we performed the FXa generation assay with various amounts of FVIII or thrombin-mediated FVIIIa, constant FIXa (1 nM), FX (300 nM) and phospholipid vesicles (PL; 20 µM) in the presence of recombinant lipidated TF (rTF, Innovin®). The Km value for FVIII in the presence of rTF was ~2.4-fold lower than that its absence (Km; 5.6±0.5, 13.3±3.7 nM, respectively, p<0.05). Similarly, the Km for FVIIIa in the presence of TF was ~1.5-fold lower than that in its absence (Km; 6.3±0.6 nM, 9.7±1.6 nM, respectively, p<0.05), supporting that the presence of TF could promote the FXa-catalyzed activation of FVIII and FVIIIa-dependent generation of intrinsic FXa. To further evaluate the effect of TF on FVIII-dependent FXa generation, the FXa generation assay with FVIIa/TF-activated FVIIIa (FVIIIa-VIIa/TF) was also performed. The initial velocity on FXa generation with FVIIIa-VIIa/TF was 22.6 nM/min. However, the initial velocity on FXa generation with FVIIIa-VIIa/TF by addition of FVIIa-inhibitor (E-76), not to generate FVIIa/TF-dependent FXa, was 3.4 nM/min, and that with FVIII alone was 0.05 nM/min. In addition, the initial velocity with FVIIa/TF alone was 10.4 nM/min. These findings supported that the TF increased FXa generation greater than the additive effect of FVIII-dependent and FVIIa/TF-dependent FXa generation in early initiation phase of coagulation prior to thrombin generation. Since tissue factor pathway inhibitor (TFPI) is present in physiological circulating whole blood, a similar experiment on FXa generation assay was repeated under the presence of TFPI, estimated to be present at 0.5 nM in the pre-coagulant state or at 15 nM in the coagulant state in circulating blood. The initial velocity on FXa generation with FVIIIa-VIIa/TF was reduced by the presence of 0.5 or 15 nM TFPI (17.9 and 12.6 nM/min, respectively). By contrast, the initial velocity on FVIIIa-VIIa/TF-dependent FXa generation with addition of FVIIa inhibitor was little reduced by 0.5 nM TFPI, whilst was reduced by 15 nM TFPI (3.5 and 2.5 nM/min, respectively). These findings supported that the TFPI possibly didn’t inhibit TF on the enhanced intrinsic tenase on association with FVIII in the pre-coagulant state.We further reported that TF enabled FXa to activate FVIII, irrespective of von Willebrand factor (VWF), and the direct association of rTF and non-lipidated TF with FVIII (Furukawa, ISTH, 2015). Since TF is transmembrane protein, however, we performed a surface plasmon resonance (SPR)-based assay (BIAcore®) and solid phase-based ELISA to identify the interactive region(s) on FVIII to recombinant soluble TF (sTF; Altor BioScience), a portion of TF outside of the PL membrane. An SPR-based assay revealed the direct binding of intact FVIII, LCh (a3-A3C1C2, A3C1C2) subunit, C2 domain to immobilized sTF (Kd; 2.3±0.6, 5.8±1.0, 10.5±3.5, 11.8±0.5 nM, respectively). The intact HCh, A1 or A2 domain to sTF failed to bind, however. A non-equilibrium ELISA also revealed that sTF bound to immobilized C2 domain with moderate affinity (Kdapp; 16.9±2.2 nM), and the interaction was dependent on ionic strength and Ca2+. In addition, the presence of VWF significantly competitively inhibited the C2 and sTF binding by ~90% (IC50; 5.7 µg/ml) at the maximal concentration employed, suggesting that the C2 domain-TF interaction could activate FVIII by FXa even in the presence of VWF.We concluded that it might be possible that TF enhanced the FVIII-mediated FXa generation by not only FVIIa but also FXa, additionally this enhancing mechanism might not be suppressed by TFPI in the initiation phase of coagulation. Furthermore, TF might function to FVIII activation, irrespective of presence of VWF, by the binding to C2 domain through the competition with VWF. DisclosuresNogami:Sysmex Corporation: Patents & Royalties, Research Funding; F. Hoffmann-La Roche Ltd.: Honoraria, Membership on an entity’s Board of Directors or advisory committees; Chugai Pharmaceutical Co., Ltd.: Honoraria, Membership on an entity’s Board of Directors or advisory committees, Patents & Royalties, Research Funding. Shima:Sysmex Corporation: Patents & Royalties, Research Funding; F. Hoffmann-La Roche Ltd.: Consultancy, Honoraria, Membership on an entity’s Board of Directors or advisory committees; Chugai Pharmaceutical Co., Ltd.: Consultancy, Honoraria, Membership on an entity’s Board of Directors or advisory committees, Patents & Royalties, Research Funding.

Aberrant Expression of TIMP-2 and PBEF Genes in the Placentae of Cloned Mice Due to Epigenetic Reprogramming Error.

Cloned mice derived from somatic or ES cells show placental overgrowth (placentomegaly) at term. We had previously analyzed cloned and normal mouse placentae by using two-dimensional gel electrophoresis and mass spectrometry to identify differential protein expression patterns. Cloned placentae showed upregulation of tissue inhibitor of metalloproteinase-2 (TIMP-2), which is involved in extracellular matrix degradation and tissue remodeling, and downregulation of pre-B cell colony enhancing factor 1 (PBEF), which inhibits apoptosis and induces spontaneous labor. Here, we used Western blotting to further analyze the protein expression levels of TIMP-2 and PBEF in cloned placentae derived from cumulus cells, TSA-treated cumulus cells, intracytoplasmic sperm injection (ICSI), and natural mating (NM control). Cloned and TSA-treated cloned placentae had higher expression levels of TIMP-2 compared with NM control and ICSI-derived placentae, and there was a positive association between TIMP-2 expression and the placental weight of cloned mouse concepti. Conversely, PBEF protein expression was significantly lower in cloned and ICSI placentae compared to NM controls. To examine whether the observed differences were due to abnormal gene expression caused by faulty epigenetic reprogramming in clones, we investigated DNA methylation and histone modification in the promoter regions of the genes encoding TIMP-2 and PBEF. Sodium bisulfite sequencing did not reveal any difference in DNA methylation between cloned and NM control placentae. However, ChIP assays revealed that the level of H3-K9/K14 acetylation at the TIMP-2 locus was higher in cloned placentae than in NM controls, whereas acetylation of the PBEF promoter was lower in cloned and ICSI placenta versus NM controls. These results suggest that cloned placentae appear to suffer from failure of histone modification-based reprogramming in these (and potentially other) developmentally important genes, leading to aberrant expression of their protein products. These changes are likely to be involved in generating the abnormalities seen in cloned mouse placentae, including enlargement and/or a lack of proper placental function.

Abstract 12475: Mechanisms of Akt1-mediated Hypothermia Protection After Cardiac Arrest: Metabolism, Inflammation and Contractile Function

Introduction: Induction of intra-arrest cooling (IC) improves cardiac, neurological and survival outcomes in preclinical models of cardiac arrest. Previous studies demonstrate that IC protection is mediated by Akt1 activation through initiating survival signaling. Hypothesis: We hypothesize that IC-induced Akt1 activation modulates protective calcium cycling proteins, metabolic, and inflammatory pathways. Methods: Adult female Akt1 +/- underwent 8 min of KCl-induced asystolic arrest and were randomized to receive IC (30 ± 0.5°C) or normothermia (NT) before CPR (n = 10 per group). 72 h survival and mean blood pressure (MAP) were measured. Indicators of glucose utilization, inflammatory responses and contractile function in heart and brain of Akt1 +/+ and Akt1 +/- mice were assessed (n=5 per group). Results: No difference on survival was seen for IC and NT treated Akt1 +/- mice. IC induced transient improvement in MAP at 30 min following resuscitation (53.6 ± 2.2 vs 71.2 ± 2.6 mmHg p&lt;0.05) and increased phospholamban phosphorylation in the heart of Akt1 +/+ mice which were not seen in Akt1 +/- mice. Compared to IC-treated Akt1 +/+ , pyruvate dehydrogenase phosphorylation and sorbitol accumulation were significantly increased, and NAD and ATP contents were markedly reduced in the hearts and brains of Akt1 +/- mice. IC significantly inhibited IκBα degradation and NF-κB nuclear translocation, and reduced concentrations of blood lactate and Pre-B cell Colony Enhancing Factor in Akt1 +/+ mice, but not Akt1 +/- mice. Conclusion: Akt1 is a critical mediator for TH protection during early post resuscitation in heart and brain through regulating calcium regulatory proteins, enhancing glucose utilization and downregulating inflammatory responses.

LEF1 Overexpression Is Associated with Preneoplastic and Neoplastic Glands of the Endometrium

Objectives: Lymphoid enhancing factor 1 (LEF1) is part of Wnt/CTNNB1 signaling pathways and plays an important role in endometrial gland formation. Overexpression of LEF1 has been detected in tumors from multiple organs, specifically in solid-pseudopapillary neoplasm of the pancreas due to CTNNB1 mutation, which encodes β-catenin. Mutations of CTNNB1 are also found in approximately 12%-25% of endometrial carcinomas. This study …

Thermoelectricity in periodic and quasiperiodically segmented nanobelts and nanowires

ABSTRACTThermoelectric properties of segmented nanowires and nanobelts are studied by means of the Kubo-Greenwood formula and a real-space renormalization plus convolution method. The tight-binding and Born models are respectively used for the calculation of electronic and lattice thermal conductivities. In particular, we investigate the thermoelectric figure of merit (ZT) of periodic and quasiperiodically segmented nanowires with two different cross sections, where the segments of the quasiperiodic one are ordered following the Fibonacci sequence. The results show an increase of ZT when the cross section area of nanowires diminishes. In addition, we present results of ZT in segmented nanobelts with an inhomogeneous cross section. For both nanowires and nanobelts, the quasiperiodicity seems to be an important enhancing factor of ZT.

PID Parameters Prediction Using Neural Network for A Linear Quarter Car Suspension Control

Providing control for suspension systems in vehicles is an enhancing factor for comfort and safety. With the improvement of control conditions, it is possible to design a cost-efficient controller which will maintain optimum comfort within harsher environmental conditions. The aim of this study is to design an adaptive PID controller with a predictive neural network model, which will be referred as NPID (NeuralPID), to control a suspension system. For this purpose, a NN (Neural Network) model is designed to produce outputs for PID’s Proportional (P) parameter to provide optimum responses for different road inputs. Also, reliability of the system outputs, which is using adaptive Proportional parameter, is tested. PID parameters for linear quarter vehicle model are decided through Zeigler-Nichols method. An ideal PID model, where Integral (I) and Derivative (D) parameters are bound to Proportional parameter, is used in the system. When the outputs of different controlled and not controlled systems, which are free, PID and NPID, are compared; it has been seen that NPID outputs are more convenient. In addition, it is possible to design controllers, with adaptively adjusting P parameter, which are operating cost-effective.

Heat transfer enhancement in a closed thermosyphon with thermally conductive PA6/water

A three-phase closed thermosyphon (THPCT) was constructed with the media of thermally conductive polyamide 6 (PA6) particles and pure water. Experimental investigations on the effects of filling ratio (30%–60%), heat flux (3.96–31.70 kW⋅m−2), and solid holdup (0%–50%) were conducted to study the heat transfer performance. Results of the THPCT reveal an enhanced heat transfer performance in contrast to that of a two-phase closed thermosyphon (TPCT) using pure water. The evaporator convective heat transfer coefficient for different solid holdups is investigated to evaluate thermal performance. The enhancing factor indicates a maximum improvement of the heat transfer coefficient by 25.6%, which corresponds to 442 W⋅m−2⋅°C−1. Thermal performance is also improved in terms of the overall thermal resistance because of the addition of PA6 particles. Moreover, fluidization of solid particles is closely related to the heat transfer performance.

Thermal Denaturation and Domain Stability of NAMPT Protein

Nicotinamide phosphoribosyltransferase (NAMPT) is also known as Pre-B cell colony enhancing factor (PBEF) or visceral fat protein (Visfatin). It is a highly conserved 52-kDa protein, found in living species from bacteria to humans [1], and acts as a rate-limiting step in the salvage pathway of nicotinamide adenine dinucleotide (NAD) biosynthesis. NAMPT is involved in several inflammatory deceases notably, sepsis, acute lung injury, and rheumatoid arthritis [2]. It is also known to activate insulin receptor causing type II diabetes [3]. Because of such medicinal relevance, NAMPT has been extensively investigated by X-ray crystallography, aiming at developing inhibitors [1]. However, physiologically relevant biochemical characterization including nuclear magnetic resonance (NMR) spectroscopy for NAMPT is scarce. NMR methods are complementary to X-ray crystallography, and provide valuable information on the structure and functional dynamics simultaneously. They offer an important tool for understanding biological functioning [4]. Bacterial expression, biochemical characterization, and thermal stability studies on NAMPT have been undertaken with the objective of optimizing conditions for NMR experiments. Secondary structure estimates using circular dichroism (CD) spectroscopy are consistent with crystallographic data. Thermal unfolding studies indicated an unusually high degree of thermal stability of NAMPT showing a melting temperature of about 90 °C. Results for dynamic light scattering (DLS) experiments showed a salt-dependent multimeric state. Presently, we are optimizing solvent conditions for obtaining the stable oligomeric state relevant to NMR experiments. Calorimetric studies are also under progress. We successfully expressed NAMPT in E.coli and are developing strategies for better yields and isotope labelling. [1] T. Wang et al. (2006) Nat. Struct. Mol. Biol. 13, 661. [2] S. M. Camp et al. (2015) Sci. Rep. 5, 13135. [3] A. Fukuhara et al. (2005) Science 307, 426. [4] T. R. Molugu et al. (2015) Concepts Magn. Reson. (in press).

Strong association of the polymorphisms in PBEF1 and knee OA risk: a two-stage population-based study in China.

The association of Pre-B cell colony enhancing factor 1 (PBEF1) with obesity, together with its pro-inflammatory properties suggests that PBEF1 might be another crucial mediator that links inflammation with obesity and primary osteoarthritis (OA). We hypothesized that polymorphisms in PBEF1 may modify the risk of developing OA. Thus we systematically screened 4 tagging polymorphisms (rs4730153, rs2058540, rs3801267 and rs16872158) in PBEF1 and evaluated the association between the genetic variants and OA risk in a two-stage case-control study including 196 cases and 442 controls in the first stage and 143 cases and 238 controls in the second stage. In the first stage, two SNPs (rs4730153 and rs16872158) were found to be potentially associated with OA risk (P < 0.05), which were further confirmed in the second stage with similar effects. After combining the two stages, we found that rs4730153 was significantly associated with decreased risk of OA in an additive genetic model (P < 0.05), while rs16872158 showed increased risk of developing OA (P < 0.05). Combined analysis of these 2 SNPs showed a significant allele-dosage association between the number of risk alleles and OA risk (Ptrend = 5.25 × 10−5). These findings indicate that genetic variants in PBEF1 gene may modify individual susceptibility to OA in the Chinese population.

Amyloid persistence in decellularized liver: biochemical and histopathological characterization

Systemic amyloidoses are a group of debilitating and often fatal diseases in which fibrillar protein aggregates are deposited in the extracellular spaces of a range of tissues. The molecular basis of amyloid formation and tissue localization is still unclear. Although it is likely that the extracellular matrix (ECM) plays an important role in amyloid deposition, this interaction is largely unexplored, mostly because current analytical approaches may alter the delicate and complicated three-dimensional architecture of both ECM and amyloid. We describe here a decellularization procedure for the amyloidotic mouse liver which allows high-resolution visualization of the interactions between amyloid and the constitutive fibers of the extracellular matrix. The primary structure of the fibrillar proteins remains intact and the amyloid fibrils retain their amyloid enhancing factor activity.