Fas Activation Research Articles

Expression of p53 in human adipose tissue correlates positively with FAS and BMI.

Activation of Fas (CD95) in adipocytes inhibits browning and may contribute to body weight gain in mice. Moreover, Fas expression in white adipose tissue (WAT) correlates positively with body mass index (BMI) in humans. However, molecular pathways involved in the inhibitory effect of Fas on energy metabolism remain incompletely understood. Herein, we report that protein levels of the tumor suppressor p53 were reduced in primary white adipocytes of adipocyte-specific Fas-knockout mice. Moreover, Fas ligand (FasL) treatment increased p53 concentrations in cultured adipocytes and decreased mitochondrial oxygen consumption in control but not in p53-depleted cells, indicating that Fas activation reduces energy expenditure in a p53-dependent manner. In line, in differentiated human mesenchymal stem cells and WAT derived from different anatomical depots, FAS expression was positively associated with p53. Furthermore, p53 expression in human subcutaneous and visceral WAT correlated positively with BMI, whereas its expression in visceral WAT was inversely associated with insulin sensitivity (as assessed by hyperinsulinemic-euglycemic clamp). Taken together, our data suggest that Fas regulates p53 expression in adipocytes, and may thereby affect body weight gain and insulin sensitivity.

Fas (CD95) expression in adipocytes contributes to diet-induced obesity.

Induction of browning in white adipose tissue (WAT) increases energy expenditure and may be an attractive target for the treatment of obesity. Since activation of Fas (CD95) induces pathways known to blunt expression of uncoupling protein 1 (UCP1), we hypothesized that Fas expression in adipocytes inhibits WAT browning and thus contributes to the development of obesity. Adipocyte-specific Fas knockout (FasΔadipo) and control littermate (FasF/F) mice were fed a regular chow diet or a high-fat diet (HFD) for 20 weeks. Energy expenditure was assessed by indirect calorimetry, and browning was determined in subcutaneous WAT. In vitro, UCP1 was analyzed in subcutaneous murine adipocytes treated with or without Fas ligand. Moreover, FAS expression in WAT was correlated to UCP1 and percentage of body fat in human individuals. HFD-fed FasΔadipo mice displayed reduced body weight gain and blunted adiposity compared to control littermates. Concomitantly, whole-body energy expenditure and WAT browning were elevated. In cultured adipocytes, Fas ligand treatment blunted isoproterenol-induced UCP1 protein levels. In support of these findings in rodents, FAS expression in WAT correlated negatively with UCP1 but positively with adiposity in human individuals. Fas activation in adipocytes contributes to HFD-associated adiposity in rodents and may be a therapeutic target to reduce obesity and associated diseases.

Comparative effects of viable Lactobacillus rhamnosus GG and its heat-inactivated paraprobiotic in the prevention of high-fat high-fructose diet-induced non-alcoholic fatty liver disease in rats.

Nonalcoholic fatty liver disease (NAFLD) is one of the most prevalent chronic liver alterations worldwide, being gut microbiota dysbiosis one of the contributing factors to its development. The aim of this research is to compare the potential effects of a viable probiotic (Lactobacillus rhamnosus GG) with those exerted by its heat-inactivated paraprobiotic counterpart in a dietary rodent model of NAFLD. The probiotic administration effectively prevented the hepatic lipid accumulation induced by a high-fat high-fructose diet feeding, as demonstrated by chemical (lower TG content) and histological (lower steatosis grade and lobular inflammation) analyses. This effect was mainly mediated by the downregulation of lipid uptake (FATP2 protein expression) and upregulating liver TG release to bloodstream (MTTP activity) in rats receiving the probiotic. By contrast, the effect of the paraprobiotic preventing diet-induced liver lipid accumulation was milder, and mainly derived from the downregulation of hepatic de novo lipogenesis (SREBP-1c protein expression and FAS activity) and TG assembly (DGAT2 and AQP9 protein expression). The obtained results demonstrate that under these experimental conditions, the effects induced by the administration of viable L. rhamnosus GG preventing liver lipid accumulation in rats fed a diet rich in saturated fat and fructose differ from those induced by its heat-inactivated paraprobiotic counterpart.

Preservation of retinal structure and function in two mouse models of inherited retinal degeneration by ONL1204, an inhibitor of the Fas receptor

Due to the large number of genes and mutations that result in inherited retinal degenerations (IRD), there has been a paucity of therapeutic options for these patients. There is a large unmet need for therapeutic approaches targeting shared pathophysiologic pathways in a mutation-independent manner. The Fas receptor is a major activator and regulator of retinal cell death and inflammation in a variety of ocular diseases. We previously reported the activation of Fas-mediated photoreceptor (PR) cell death in two different IRD mouse models, rd10 and P23H, and demonstrated the protective effect of genetic Fas inhibition. The purpose of this study was to examine the effects of pharmacologic inhibition of Fas in these two models by intravitreal injection with a small peptide inhibitor of the Fas receptor, ONL1204. A single intravitreal injection of ONL1204 was given to one eye of rd10 mice at P14. Two intravitreal injections of ONL1204 were given to the P23H mice, once at P14 and again at 2-months of age. The fellow eyes were injected with vehicle alone. Fas activation, rate of PR cell death, retinal function, and the activation of immune cells in the retina were evaluated. In both rd10 and P23H mice, ONL1204 treatment resulted in decreased number of TUNEL (+) PRs, decreased caspase 8 activity, enhanced photoreceptor cell counts, and improved visual function compared with vehicle treated fellow eyes. Treatment with ONL1204 also reduced immune cell activation in the retinas of both rd10 and P23H mice. The protective effect of pharmacologic inhibition of Fas by ONL1204 in two distinct mouse models of retinal degeneration suggests that targeting this common pathophysiologic mechanism of cell death and inflammation represents a potential therapeutic approach to preserve the retina in patients with IRD, regardless of the genetic underpinning.

The Chaochou active fault in southern Taiwan: Characteristics and geomorphological significance as a reverse fault

The Chaochou Fault in southern Taiwan has long been recognized as an active fault, but its exact location is still uncertain. In this study, we focused on the middle part of the Chaochou Fault, where a flight of fluvial terraces was deformed and preserved. High-resolution aerial photos were first used to observe and map these terraces and their vicinity, followed by comprehensive field investigations including RTK-GPS surveys for high-resolution topographic profiles of the deformed terraces and OSL sample collections for obtaining the ages of the terraces. A series of active faults, consisting of Fault A (“FA”), Fault B (“FB”), and Fault Bb (“FBb”), are collectively named the Chaochou Active Fault Zone. The fault zone is recognized and characterized by range-facing scarps, frontal flexural scarps, and discontinuous slopes deformed by FA, FB, and FBb, respectively. The repeated activities of FA were recorded by the incremental fault scarp heights from young to old terraces. Considering the highest fault scarp height of 65.9 ± 6 m observed in T1 and its OSL age of 35.3 ± 4.3 ka, the long-term fault slip rate of FA is about 2 mm/yr.

A molecular switch controls assembly of bacterial focal adhesions.

Cell motility universally relies on spatial regulation of focal adhesion complexes (FAs) connecting the substrate to cellular motors. In bacterial FAs, the Adventurous gliding motility machinery (Agl-Glt) assembles at the leading cell pole following a Mutual gliding-motility protein (MglA)-guanosine 5'-triphosphate (GTP) gradient along the cell axis. Here, we show that GltJ, a machinery membrane protein, contains cytosolic motifs binding MglA-GTP and AglZ and recruiting the MreB cytoskeleton to initiate movement toward the lagging cell pole. In addition, MglA-GTP binding triggers a conformational shift in an adjacent GltJ zinc-finger domain, facilitating MglB recruitment near the lagging pole. This prompts GTP hydrolysis by MglA, leading to complex disassembly. The GltJ switch thus serves as a sensor for the MglA-GTP gradient, controlling FA activity spatially.

Abstract PO2-15-04: Fas/FasL expression on circulating tumor and immune cells in the peripheral blood of patients with metastatic breast cancer (BC): correlation with clinical outcome

Abstract Background: The activation of Fas (CD95/APO-1) death receptor by Fas ligand (FasL/CD95L) can trigger a signal transduction pathway leading to apoptosis, which is a common pathway used by immune cells for tumor elimination. Cancer cells also exploit FasL expression to induce immune cell apoptosis, known as "Fas counterattack", and thus to increase their invasion and migration capacity. We herein assessed the expression of Fas and FasL on circulating tumor cells (CTCs) and immune cells in the peripheral blood (PB) of patients with metastatic BC. Methods: PB was obtained from 98 patients with metastatic BC at baseline before first-line treatment. Peripheral blood mononuclear cells (PBMCs) were isolated and immunofluorescently stained with antibodies against cytokeratins (CK; clones: AE1/AE3 & C11) and CD45 for CTC detection, along with antibodies for Fas, FasL and dapi. Fas/FasL expression was individually assessed on CTCs and PBMCs using the Ariol microscopy system (2x10^6 PBMCs were analyzed per patient). Results: CTCs were detected in 26/98 (26.5%) patients (total CTC count: n=70). Fas+ CTCs and FasL+ CTCs were identified in 88.5% and 92.3% of CTC-positive patients, respectively, representing the 57.1% and 82.9% of total CTCs. Regarding co-expression at single cell level, Fas+/FasL+ CTCs were detected in 84.6% of patients, whereas Fas+/FasL- CTCs, Fas-/FasL+ CTCs and Fas-/FasL- CTCs were identified in 7.7%, 19.2% and 11.5% of patients, respectively. Regarding PBMCs, the Fas+/FasL+ phenotype was identified in 70.3%, of patients, whereas Fas-/FasL+ PBMCs and Fas-/FasL- PBMCs were detected in 24.2% and 5.5% of patients, respectively; interestingly the Fas+/FasL- expression pattern was not observed in any patient. A reduced progression-free survival (PFS) was demonstrated among CTC-positive as compared to CTC-negative patients (median PFS: 9.5 versus 13.4 months; p=0.004), and specifically those harboring Fas+/FasL+ CTCs (median PFS: 9.5 vs 13.4 months; p=0.009). Increased overall survival (OS) was demonstrated among patients harboring Fas+/FasL+ PBMCs, as compared to those with Fas-/FasL+ PBMCs and Fas-/FasL- PBMCs (median OS: 35.7 vs 25.9 vs 14.4 months; p=0.008). Conclusion: Herein we provide for the first time evidence on Fas/FasL expression on CTCs and PBMCs with significant prognostic relevance for patients with metastatic BC. The findings highlight the Fas/FasL signaling pathway as a putative mechanism of immune evasion and metastatic progression of BC. Their role as prognostic biomarkers in BC patients merits further investigation. Citation Format: Maria Papadaki, Maria Vassilakopoulou, Eleni Papadaki, Despoina Aggouraki, Chrysostomos Zioudas, Anastasia Mala, Sofia Agelaki, Dimitrios Mavroudis. Fas/FasL expression on circulating tumor and immune cells in the peripheral blood of patients with metastatic breast cancer (BC): correlation with clinical outcome [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-15-04.

Synthesis of geopolymers using municipal solid waste incineration fly ash and construction and demolition waste: Mechanical and thermal activation

The safe disposal and utilization of MSWI FA, enriching with high amounts of heavy metals, chlorides and organic pollutants, has turned into a critical problem that restricts MSWI development. In this study, the resource utilization feasibility of MSWI FA with CDW and GGFBs by geopolymers was analyzed, and the performance of geopolymers with the different MSWI FA dose and activation method was investigated. With the increase of MSWI FA addition, the compressive strength of geopolymer showed a trend of first increasing and then decreasing. Mechanical and thermal activation had a significant effect on the properties of geopolymers through improving the reactivity of raw materials and promoting hydration reaction. Various characterization methods indicated hydration products such as CSH, CASH and NASH gel were generated in geopolymers. In terms of S/S of pollutants, geopolymers exhibited excellent capability. Compared with MSWI FA, the concentration of heavy metals, especially Pb, in the leachate from geopolymers was significantly reduced due to the transformation of heavy metals into more stable forms. Moreover, mechanical and thermal activation could promote the S/S of heavy metals, except Pb and Zn. After geopolymerization, the total TEQ of PCDD/Fs decreased from 42.8 to 4.7 ng TEQ/kg with the IE of 45.09%. This article provides a new method for the collaborative resource utilization of MSWI FA, CDW and GGFBs.

Intermediate Phase Engineering with 2,2-Azodi(2-Methylbutyronitrile)for Efficient and Stable Perovskite Solar Cells.

Sequential deposition has been widely employed to modulate the crystallization of perovskite solar cells because it can avoid the formation of nucleation centers and even initial crystallization in the precursor solution. However, challenges remain in overcoming the incomplete and random transformation of PbI2 films with organic ammonium salts. Herein, a unique intermediate phase engineering strategy has been developed by simultaneously introducing 2,2-azodi(2-methylbutyronitrile) (AMBN) to both PbI2 and ammonium salt solutions to regulate perovskite crystallization. AMBN not only coordinates with PbI2 to form a favorably mesoporous PbI2 film due to the coordination between Pb2+ and the cyano group (C≡N), but also suppresses the vigorous activity of FA+ ions by interacting with FAI, leading to the full PbI2 transformation with the preferred orientation. Therefore, perovskites with favorable facet orientations are obtained, and the defects are largely suppressed owing to the passivation of uncoordinated Pb2+ and FA+ . As a result, a champion power conversion efficiency over 25% with a stabilized efficiency of 24.8% is achieved. Moreover, the device exhibits an improved operational stability, retaining 96% of initial power conversion efficiency under 1000 h continuous white-light illumination with an intensity of 100mW cm-2 at ≈55°C in N2 atmosphere.

Oral Administration of Chaetoceros gracilis—A Marine Microalga—Alleviates Hepatic Lipid Accumulation in Rats Fed a High-Sucrose and Cholesterol-Containing Diet

Microalgae are attracting attention as a next-generation alternative source of protein and essential fatty acids that do not consume large amounts of water or land. Chaetoceros gracilis (C. gracilis)-a marine microalga-is rich in proteins, fucoxanthin, and eicosapentaenoic acid (EPA). Growing evidence indicates that dietary fucoxanthin and EPA have beneficial effects in humans. However, none of these studies have shown that dietary C. gracilis has beneficial effects in mammals. In this study, we investigated the effects of dietary C. gracilis on lipid abnormalities in Sprague-Dawley rats fed a high-sucrose cholesterol-containing diet. Dried C. gracilis was added to the control diet at a final dose of 2 or 5% (w/w). After four weeks, the soleus muscle weights were found to be dose-responsive to C. gracilis and showed a tendency to increase. The hepatic triglyceride and total cholesterol levels were significantly reduced by C. gracilis feeding compared to those in the control group. The activities of FAS and G6PDH, which are related to fatty acid de novo synthesis, were found to be dose-responsive to C. gracilis and tended to decrease. The hepatic glycerol content was also significantly decreased by C. gracilis feeding, and the serum HDL cholesterol levels were significantly increased, whereas the serum levels of cholesterol absorption markers (i.e., campesterol and β-sitosterol) and the hepatic mRNA levels of Scarb1 were significantly decreased. Water-soluble metabolite analysis showed that the muscular contents of several amino acids, including leucine, were significantly increased by C. gracilis feeding. The tendency toward an increase in the weight of the soleus muscle as a result of C. gracilis feeding may be due to the enhancement of muscle protein synthesis centered on leucine. Collectively, these results show that the oral administration of C. gracilis alleviates hepatic lipid accumulation in rats fed a high-sucrose and cholesterol-containing diet, indicating the potential use of C. gracilis as a food resource.

Increasing Bioavailability of Trans-Ferulic Acid by Encapsulation in Functionalized Mesoporous Silica

Two types of mesoporous materials, MCM-41 and MCM-48, were functionalized by the soft-template method using (3-aminopropyl)triethoxysilane (APTES) as a modifying agent. The obtained mesoporous silica materials were loaded with trans-ferulic acid (FA). In order to establish the morphology and structure of mesoporous materials, a series of specific techniques were used such as: X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM), Brunauer-Emmet-Teller (BET), Fourier Transform Infrared Spectroscopy (FTIR) and thermogravimetric analysis (TGA). We monitored the in vitro release of the loaded FA at two different pH values, by using simulated gastric fluid (SGF) and simulated intestinal fluid (SIF). Additionally, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and Candida albicans ATCC 10231 were used to evaluate the antimicrobial activity of FA loaded mesoporous silica materials. In conclusion such functionalized mesoporous materials can be employed as controlled release systems for polyphenols extracted from natural sources.

Role of let-7 family in the invasion and metastasis of osteosarcoma.

Role of let-7 family in the invasion and metastasis of osteosarcoma.

Maternal hyperthyroidism increases the synthesis activity and the osteogenic markers expression of calvarial osteoblasts from offspring in a murine model

To evaluate the characteristics and synthesis activity of osteoblasts extracted from the calvaria of offspring of rats exposed to maternal hyperthyroidism. Twelve adult Wistar rats were divided into two groups, one control and one treated with daily administration of L-thyroxine by an orogastric tube (50 µg/animal/day) during pregnancy. Three days after delivery and confirmation of the mothers’ hyperthyroidism, the offspring were euthanized for the extraction of osteoblasts from the calvaria. At 7, 14, and 21 days, proliferation activity was assessed using MTT assay, while alkaline phosphatase (ALP) activity was assessed by the BCIP/NBT method. At 21 days, the total area of the mineralized matrix stained by von Kossa was evaluated by morphometry. The expression of gene transcripts for Runx2, Bmp2, Fgfr1, collagen type 1 (Col1), osteocalcin (Oc), and osteopontin (Op) were evaluated by real-time RT-PCR. Means were compared using the Student’s t-test. FA activity was significantly higher at 14 and 21 days in cultures of osteoblasts extracted from offspring exposed to maternal hyperthyroidism, while MTT conversion was significantly lower at 21 days in this group. Osteoblast cultures of neonates exposed to maternal hyperthyroidism also showed a larger total area of mineralized matrix and greater expression of gene transcripts for Oc and Op. Maternal hyperthyroidism increases the activity of matrix synthesis, alkaline phosphatase activity, and expression of gene transcripts for osteocalcin and osteopontin in the osteoblasts, extracted from the calvaria of the offspring, which may be one of the mechanisms of premature fusion of cranial sutures.

Antitumor and antimicrobial activity of fatty acids from green microalga Coelastrella sp. BGV

Green microalgae are a valuable source of biologically active substances with antitumor and antimicrobial effects. Coelastrella sp. BGV is a strain of green microalga isolated in Bulgaria. The aim of the study was to analyze the fatty acid profile of Coelastrella sp. BGV and to assess the antitumor and antimicrobial activities of the extracted free fatty acids. The fatty acid content of a total lipid extract from the green microalga Coelastrella sp. BGV was determined by gas chromatography mass spectrometry. The antitumor activity of FAs against human HeLa tumor cells was estimated by MTT testing and fluorescence microscopy methods. The antimicrobial activity of FAs against Gram-positive, Gram-negative bacteria and the fungal pathogen Candida albicans was studied by the disk diffusion and microdilution methods. Eleven fatty acids were identified in the lipid extract of Coelastrella sp. BGV, of which the monounsaturated oleic acid (18:1) was present in the largest amount - 31.7%, followed by the polyunsaturated linoleic acid (18:2) - 25.9% and saturated palmitic acid (16:0) - 19.2%. The ratio of total unsaturated to saturated fatty acids was 3:1, and the monounsaturated to polyunsaturated fatty acid ratio was 1:1. The extracted FAs showed statistically significant antiproliferative activity against HeLa tumor cells and induced morphological changes in the cell and nucleus characteristic of early and/or late apoptosis. FAs showed varying degrees of antibacterial and antifungal activity against Gram-positive, Gram-negative pathogens and C. albicans . The results obtained are promising in terms of the therapeutic potential of FAs from Coelastrella sp. BGV against tumor cells and against Gram-negative bacteria ( Escherichia coli UPEC, Pseudomonas aeruginosa, Klebsiella pneumoniae ) and the fungal pathogen Candida albicans . • The fatty acid profile of green microalga strain Coelastrella sp. BGV was analyzed. • The antitumor and antimicrobial effects of the isolated fatty acids was assessed. • The lipid extract had a high content of unsaturated fatty acids. • The fatty acids inhibited the growth of HeLa tumor cells and induced apoptosis. • The fatty acids had antimicrobial and antifungal activity.

Gryllus bimaculatus extract protects against palmitate-induced β-cell death by inhibiting ceramide synthesis

Type I diabetes mellitus is an autoimmune disease characterized by the destruction of β-cells, leading to severe insulin deficiency. Environmental factors and genetic predisposition are implicated in β-cell destruction, which is the final step in a cascade of complex events. Possible triggers of β-cell destruction are activation of Fas, activation of perforin, increased generation of reactive oxygen species, increased production of inflammatory cytokines, and endoplasmic reticulum (ER) stress. In this study, we examined whether Gryllus bimaculatus (GB) extract could prevent palmitate-induced β-cell apoptosis. Exposure to GB extract prevented palmitate-induced death of MIN6 cells, a mouse pancreatic β-cell line. Palmitate increased total ceramide levels with the elevation of ceramide synthase (CerS)1, CerS4, and CerS6 expressions. Treatment with GB extract decreased the levels and expressions of ceramides related to insulin resistance. CerS4 and CerS6 overexpression, but not CerS1 overexpression, increased palmitate-induced MIN6 cell death by increasing ceramide synthesis. Oppositely, inhibition of ceramide synthesis by fumonisin B1 treatment partially recovered palmitate-induced MIN6 cell death. Furthermore, GB extract reduced ER stress (phosphorylation of PERK and eIF2α), NF-κB–iNOS signaling, and the phosphorylation of MAP kinase (JNK, p38). GB extract reduced pro-apoptotic Bax protein expression but increased anti-apoptotic Bcl2 expression. In addition, CerS4 and CerS6 overexpression aggravated impairment of insulin secretion by palmitate, but GB extract recovered it. In conclusion, GB could be a functional food that improves palmitate-induced β-cell death and insulin secretion.

Carboxypeptidase G and pterin deaminase metabolic pathways degrade folic acid in Variovorax sp. F1

BackgroundFolic acid (FA) is a synthetic vitamin (B9) and the oxidized form of a metabolic cofactor that is essential for life. Although the biosynthetic mechanisms of FA are established, its environmental degradation mechanism has not been fully elucidated. The present study aimed to identify bacteria in soil that degrade FA and the mechanisms involved.ResultsWe isolated the soil bacterium Variovorax sp. F1 from sampled weed rhizospheres in a grassland and investigated its FA degradation mechanism. Cultured Variovorax sp. F1 rapidly degraded FA to pteroic acid (PA), indicating that FA hydrolysis to PA and glutamate. We cloned the carboxypeptidase G (CPG) gene and found widely distributed paralogs within the Variovorax genus. Recombinant CPG preferred FA and deaminofolic acid as substrates, indicating its involvement in FA degradation by Variovorax. Prolonged culture of Variovorax sp. F1 resulted in decreased rates of deaminofolic acid (DFA) and deaminopteroic acid (DPA) accumulation. This indicated that the deamination reaction also comprised a route of FA degradation. We also identified an F1 gene that was orthologous to the pterin deaminase gene (Arad3529) of Agrobacterium radiobacter. The encoded protein deaminated FA and PA to DFA and DPA, which was consistent with the deamination activity of FA and PA in bacterial cell-free extracts.ConclusionWe discovered that the two enzymes required for FA degradation pathways in isolates of Variovorax sp. F1 comprise CPG and pterin deaminase, and that DFA and PA are intermediates in the generation of DPA.

Functional roles for AGPAT6 in milk fat synthesis of buffalo mammary epithelial cells

AGPAT6 plays a crucial role in the triglyceride (TG) synthesis pathway in mammals. However, its roles in buffalo lactation remain unknown. Therefore, we investigated the functional roles of AGPAT6 in milk fat synthesis by transfecting overexpression and lentivirus interference vectors in buffalo mammary epithelial cells (BuMECs) in vitro. AGPAT6 overexpression in BuMECs significantly enhanced the mRNA expression of FABP4, SLC27A6, ACSL1, DGAT1, DGAT2, LPIN1, INSIG1, CEBPA and SREBF1 genes, and significantly reduced that of XDH, CPT1A, LIPE, INSIG2 and PPARGC1A, but has no significant influence to the mRNA abundance of FABP3, GPAM, PPARG and SREBF2. However, the interference with AGPAT6, the mRNA expression of FABP4, SLC27A6, ACSL1, DGAT1, DGAT2, INSIG1, CEBPA, SREBF1, XDH, CPT1A, LIPE, INSIG2 and PPARGC1A genes in BuMECs changed contrary to the overexpression experiment, and that of GPAM, PPARG and SREBF2 also did not change significantly, but the expression of FABP3 was significantly decreased. In addition, the overexpression/interference of AGPAT6 gene significantly increased/decreased TG content in BuMECs. The results here indicate that AGPAT6 gene is involved in TG synthesis in BuMECs, and affects the expression of major genes associated with FA transport and activation, TG synthesis and transcription regulation, FA oxidation and TG degradation during the lipogenesis of milk.

Enhancement of compressive strength of high-volume fly ash cement paste by wet grinded cement: Towards low carbon cementitious materials

The use of high-volume fly ash (HVFA) in cement-based materials can significantly reduce the CO2 emissions. Nevertheless, slow strength development becomes the barrier for HVFA system, and the main reason is because the low alkalinity offered by Portland cement (PC) cannot efficiently activate the pozzolanic reaction of FA. In this study, wet grinding was used to process PC and obtain superfine particles, with intention to promote the hydration of PC and offer more CH for FA activation. HVFA (50% PC and 50% FA) system was designed, and 0–15% wet grinded cement (WGC) was used to replace PC. The effects of WGC on HVFA system were investigated; the CO2 emissions and costs of WGC-HVFA system were calculated. Results showed that the superfine cement with D50 = 3.17 μm was obtained by wet grinding. The compressive strength of HVFA mortars without WGC was 14.3 MPa and 33.5 MPa at 3 d and 28 d age, while 15% WGC enhanced the compressive strength of the mortars to 22.0 MPa and 46.6 MPa, respectively. In addition, CO2 emissions and costs of the HVFA system containing 15% WGC were 477 kg/t and 347.5 RMB/t, which were much lower than that of PC system. Such results proposed an innovative approach to enhance the compressive strength of HVFA system, with great potential in low carbon cementitious materials.

Changes in 5-Fluorouracil-induced external granular cell damage during the time-course of the developing cerebellum of infant rats.

5-Fluorouracil (5-FU) is widely used as a chemotherapeutic agent that blocks DNA synthesis and replication by inhibiting thymidylate synthetase. This study aimed to elucidate 5-FU-induced changes in the external granular cells (EGCs) in the cerebellum of infant rats and the possible underlying mechanism. Six-day-old infant rats were injected subcutaneously with 40 mg/kg of 5-FU, and their cerebellums were examined at 6, 9, 12, and 24 h after treatment (HAT), and 2, 4, and 10 d after treatment (DAT). The width of the external granular layer (EGL) decreased from 24 HAT to 4 DAT in the 5-FU group compared to that in the control group. However, the width in the 5-FU group was comparable to that of the control group at 10 DAT. The number of apoptotic cells, cleaved caspase 3-labeling index (LI%), p21cip1-LI%, and expression levels of p53, p21cip1, and Fas mRNAs increased at 24 HAT. However, no changes were detected in the expression levels of Puma and Bax mRNAs at any time point. BrdU-LI% increased at 6 and 12 HAT but decreased at 24 HAT. The phospho-histone H3-LI% decreased from 6 HAT to 2 DAT. The width of the molecular layer decreased compared to that of the control group at 10 DAT. No differences were observed in Purkinje cell development. These results indicate that 5-FU inhibited cell proliferation by inducing apoptosis of EGCs via activation of Fas and caspase-3 without the involvement of the mitochondrial pathway and induced p53-dependent G1-S and G2-M phase arrest.

NF-\u03baB regulation in maternal immunity during normal and IUGR pregnancies

Intrauterine Growth Restriction (IUGR) is a leading cause of perinatal death with no effective cure, affecting 5–10% pregnancies globally. Suppressed pro-inflammatory Th1/Th17 immunity is necessary for pregnancy success. However, in IUGR, the inflammatory response is enhanced and there is a limited understanding of the mechanisms that lead to this abnormality. Regulation of maternal T-cells during pregnancy is driven by Nuclear Factor Kappa B p65 (NF-κB p65), and we have previously shown that p65 degradation in maternal T-cells is induced by Fas activation. Placental exosomes expressing Fas ligand (FasL) have an immunomodulatory function during pregnancy. The aim of this study is to investigate the mechanism and source of NF-κB regulation required for successful pregnancy, and whether this is abrogated in IUGR. Using flow cytometry, we demonstrate that p65+ Th1/Th17 cells are reduced during normal pregnancy, but not during IUGR, and this phenotype is enforced when non-pregnant T-cells are cultured with normal maternal plasma. We also show that isolated exosomes from IUGR plasma have decreased FasL expression and are reduced in number compared to exosomes from normal pregnancies. In this study, we highlight a potential role for FasL+ exosomes to regulate NF-κB p65 in T-cells during pregnancy, and provide the first evidence that decreased exosome production may contribute to the dysregulation of p65 and inflammation underlying IUGR pathogenesis.