Chronic Exposure to Rhodobacter Sphaeroides Extract Lycogen? Prevents UVA-Induced Malondialdehyde Accumulation and Procollagen I Down-Regulation in Human Dermal Fibroblasts

UVA contributes to the pathogenesis of skin aging by downregulation of procollagen I content and induction of matrix metalloproteinase (MMP)-associated responses. Application of antioxidants such as lycopene has been demonstrated as a convenient way to achieve protection against skin aging. Lycogen™, derived from the extracts of Rhodobacter sphaeroides, exerts several biological effects similar to that of lycopene whereas most of its anti-aging efficacy remains uncertain. In this study, we attempted to examine whether Lycogen™ could suppress malondialdehyde (MDA) accumulation and restore downregulated procollagen I expression induced by UVA exposure. In human dermal fibroblasts Hs68 cells, UVA repressed cell viability and decreased procollagen I protein content accompanied with the induction of MMP-1 and MDA accumulation. Remarkably, incubation with 50 μM Lycogen™ for 24 h ameliorated UVA-induced cell death and restored UVA-induced downregulation of procollagen in a dose-related manner. Lycogen™ treatment also prevented the UVA-induced MMP-1 upregulation and intracellular MDA generation in Hs68 cells. Activation of NFκB levels, one of the downstream events induced by UVA irradiation and MMP-1 induction, were also prevented by Lycogen™ administration. Taken together, our findings demonstrate that Lycogen™ may be an alternative agent that prevents UVA-induced skin aging and could be used in cosmetic and pharmaceutical applications.     

CC16 Regulates Inflammation,ROS Generation and Apoptosis in Bronchial Epithelial Cells during Klebsiella pneumoniae Infection

Gram-negative (G-) bacteria are the leading cause of hospital-acquired pneumonia in the United States. The devastating damage caused by G- bacteria results from the imbalance of bactericidal effects and overwhelming inflammation. Despite decades of research, the underlying mechanisms by which runaway inflammation is developed remain incompletely understood. Clara Cell Protein 16 (CC16), also known as uteroglobin, is the major protein secreted by Clara cells and the most abundant protein in bronchoalveolar lavage fluid (BALF). However, the regulation and functions of CC16 during G- bacterial infection are unknown. In this study, we aimed to assess the regulation of CC16 in response to Klebsiella pneumoniae (K. pneu) and to investigate the role of CC16 in bronchial epithelial cells. After K. pneu infection, we found that CC16 mRNA expression was significantly decreased in bronchial epithelial cells. Our data also showed that K. pneu infection upregulated cytokine and chemokine genes, including IL-1β, IL-6, and IL-8 in BEAS-2B cells. Endogenously overexpressed CC16 in BEAS-2B cells provided an anti-inflammatory effect by reducing these markers. We also observed that endogenous CC16 can repress NF-κB reporter activity. In contrast, the recombinant CC16 (rCC16) did not show an anti-inflammatory effect in K. pneu-infected cells or suppression of NF-κB promoter activity. Moreover, the overexpression of CC16 reduced reactive oxygen species (ROS) levels and protected BEAS-2B cells from K. pneu-induced apoptosis.     

Homogeneous charge compression ignition of LPG and gasoline using variable valve timing in an engine

The combustion characteristics and exhaust emissions in an engine were investigated under homogeneous charge compression ignition (HCCI) operation fueled with liquefied petroleum gas (LPG) and gasoline with regard to variable valve timing (VVT) and the addition of di-methyl ether (DME). LPG is a low carbon, high octane number fuel. These two features lead to lower carbon dioxide (CO₂) emission and later combustion in an LPG HCCI engine as compared to a gasoline HCCI engine. To investigate the advantages and disadvantages of the LPG HCCI engine, experimental results for the LPG HCCI engine are compared with those for the gasoline HCCI engine. LPG was injected at an intake port as the main fuel in a liquid phase using a liquefied injection system, while a small amount of DME was also injected directly into the cylinder during the intake stroke as an ignition promoter. Different intake valve timings and fuel injection amount were tested in order to identify their effects on exhaust emissions and combustion characteristics. Combustion pressure, heat release rate, and indicated mean effective pressure (IMEP) were investigated to characterize the combustion performance. The optimal intake valve open (IVO) timing for the maximum IMEP was retarded as the λ_TOTAL was decreased. The start of combustion was affected by the IVO timing and the mixture strength (λ_TOTAL) due to the volumetric efficiency and latent heat of vaporization. At rich operating conditions, the θ_90-20 of the LPG HCCI engine was longer than that of the gasoline HCCI engine. Hydrocarbon (HC) and carbon monoxide (CO) emissions were increased as the IVO timing was retarded. However, CO₂ was decreased as the IVO timing was retarded. CO₂ emission of the LPG HCCI engine was lower than that of the gasoline HCCI engine. However, CO and HC emissions of the LPG HCCI engine were higher than those of the gasoline HCCI engine.     

An experimental investigation of CNG as an alternative fuel for a retrofitted gasoline vehicle

This paper presents test results obtained from running a 1.5 L, 4-cylinder Proton Magma retrofitted spark ignition car engine with dynamometer. Performance, fuel consumption and exhaust emissions measurements were recorded under steady state operating conditions for gasoline and compressed natural gas (CNG). The engine was converted to computer integrated bi-fueling system from a gasoline engine and was operated separately either with gasoline or CNG using an electronically controlled solenoid actuated valve system. A PC based data acquisition and control system was used for controlling all the operation. A comparative analysis of the performance and emissions has been made for gasoline and CNG. Based on the experimental results, it is transparent that CNG shows low brake mean effective pressure (BMEP), brake specific fuel consumptions (BSFC), higher efficiency and lower emissions of CO, CO₂, HC but more NO_x compared to gasoline.     

Combustion of n-butanol in a spark-ignition IC engine

Alcohols, because of their potential to be produced from renewable sources and because of their high quality characteristics for spark-ignition (SI) engines, are considered quality fuels which can be blended with fossil-based gasoline for use in internal combustion engines. They enable the transformation of our energy basis in transportation to reduce dependence on fossil fuels as an energy source for vehicles. The research presented in this work is focused on applying n-butanol as a blending agent additive to gasoline to reduce the fossil part in the fuel mixture and in this way to reduce life cycle CO₂ emissions. The impact on combustion processes in a spark-ignited internal combustion engine is also detailed. Blends of n-butanol to gasoline with ratios of 0%, 20%, and 60% in addition to near n-butanol have been studied in a single cylinder cooperative fuels research engine (CFR) SI engine with variable compression ratio manufactured by Waukesha Engine Company. The engine is modified to provide air control and port fuel injection. Engine control and monitoring was performed using a target-based rapid-prototyping system with electronic sensors and actuators installed on the engine [1]. A real-time combustion analysis system was applied for data acquisition and online analysis of combustion quantities. Tests were performed under stoichiometric air-to-fuel ratios, fixed engine torque, and compression ratios of 8:1 and 10:1 with spark timing sweeps from 18° to 4° before top dead center (BTDC). On the basis of the experimental data, combustion characteristics for these fuels have been determined as follows: mass fraction burned (MFB) profile, rate of MFB, combustion duration and location of 50% MFB. Analysis of these data gives conclusions about combustion phasing for optimal spark timing for maximum break torque (MBT) and normalized rate for heat release. Additionally, susceptibility of 20% and 60% butanol-gasoline blends on combustion knock was investigated. Simultaneously, comparison between these fuels and pure gasoline in the above areas was investigated. Finally, on the basis of these conclusions, characteristic of these fuel blends as substitutes of gasoline for a series production engine were discussed.     

Hypoxia-Induced Collagen Synthesis of Human Lung Fibroblasts by Activating the Angiotensin System

The exact molecular mechanism that mediates hypoxia-induced pulmonary fibrosis needs to be further clarified. The aim of this study was to explore the effect and underlying mechanism of angiotensin II (Ang II) on collagen synthesis in hypoxic human lung fibroblast (HLF) cells. The HLF-1 cell line was used for in vitro studies. Angiotensinogen (AGT), angiotensin converting enzyme (ACE), angiotensin II type 1 receptor (AT1R) and angiotensin II type 2 receptor (AT2R) expression levels in human lung fibroblasts were analysed using real-time polymerase chain reaction (RT-PCR) after hypoxic treatment. Additionally, the collagen type I (Col-I), AT1R and nuclear factor κappaB (NF-κB) protein expression levels were detected using Western blot analysis, and NF-κB nuclear translocation was measured using immunofluorescence localization analysis. Ang II levels in HLF-1 cells were measured with an enzyme-linked immunosorbent assay (ELISA). We found that hypoxia increased Col-I mRNA and protein expression in HLF-1 cells, and this effect could be inhibited by an AT1R or AT2R inhibitor. The levels of NF-κB, RAS components and Ang II production in HLF-1 cells were significantly increased after the hypoxia exposure. Hypoxia or Ang II increased NF-κB-p50 protein expression in HLF-1 cells, and the special effect could be inhibited by telmisartan (TST), an AT1R inhibitor, and partially inhibited by PD123319, an AT2R inhibitor. Importantly, hypoxia-induced NF-κB nuclear translocation could be nearly completely inhibited by an AT1R or AT2R inhibitor. Furthermore pyrrolidine dithiocarbamate (PDTC), a NF-κB blocker, abolished the expression of hypoxia-induced AT1R and Col-I in HLF-1 cells. Our results indicate that Ang II-mediated NF-κB signalling via ATR is involved in hypoxia-induced collagen synthesis in human lung fibroblasts.     

Reciprocal Changes in miRNA Expression with Pigmentation and Decreased Proliferation Induced in Mouse B16F1 Melanoma Cells by l-Tyrosine and 5-Bromo-2′-Deoxyuridine

Background: Many microRNAs have been identified as critical mediators in the progression of melanoma through its regulation of genes involved in different cellular processes such as melanogenesis, cell cycle control, and senescence. However, microRNAs’ concurrent participation in syngeneic mouse B16F1 melanoma cells simultaneously induced decreased proliferation and differential pigmentation by exposure to 5-Brd-2′-dU (5’Bromo-2-deoxyuridine) and L-Tyr (L-Tyrosine) respectively, is poorly understood. Aim: To evaluate changes in the expression of microRNAs and identify which miRNAs in-network may contribute to the functional bases of phenotypes of differential pigmentation and reduction of proliferation in B16F1 melanoma cells exposed to 5-Brd-2′-dU and L-Tyr. Methods: Small RNAseq evaluation of the expression profiles of miRNAs in B16F1 melanoma cells exposed to 5-Brd-2′-dU (2.5 μg/mL) and L-Tyr (5 mM), as well as the expression by qRT-PCR of some molecular targets related to melanogenesis, cell cycle, and senescence. By bioinformatic analysis, we constructed network models of regulation and co-expression of microRNAs. Results: We confirmed that stimulation or repression of melanogenesis with L-Tyr or 5-Brd-2′-dU, respectively, generated changes in melanin concentration, reduction in proliferation, and changes in expression of microRNAs 470-3p, 470-5p, 30d-5p, 129-5p, 148b-3p, 27b-3p, and 211-5p, which presented patterns of coordinated and reciprocal co-expression, related to changes in melanogenesis through their putative targets Mitf, Tyr and Tyrp1, and control of cell cycle and senescence: Cyclin D1, Cdk2, Cdk4, p21, and p27. Conclusions: These findings provide insights into the molecular biology of melanoma of the way miRNAs are coordinated and reciprocal expression that may operate in a network as molecular bases for understanding changes in pigmentation and decreased proliferation induced in B16F1 melanoma cells exposed to L-Tyr and 5-Brd-2′-dU.     

An experimental study for the effects of boost pressure on the performance and exhaust emissions of a DI-HCCI gasoline engine

As an alternative combustion mode, the HCCI combustion has some benefits compared to conventional SI and CI engines, such as low NO_x emission and high thermal efficiency. However, this combustion mode can produce higher UHC and CO emissions than those of conventional engines. In the naturally aspirated HCCI engines, the low engine output power limits its use in the current engine technologies. Intake air pressure boosting is a common way to improve the engine output power which is widely used in high performance SI and CI engine applications. Therefore, in this study, the effect of inlet air pressure on the performance and exhaust emissions of a DI-HCCI gasoline engine has been investigated after converting a heavy-duty diesel engine to a HCCI direct-injection gasoline engine. The experiments were performed at three different inlet air pressures while operating the engine at the same equivalence ratio and intake air temperature as in normally aspirated HCCI engine condition at different engine speeds. The SOI timing was set dependently to achieve the maximum engine torque at each test condition. The effects of inlet air pressure both on the emissions such as CO, UHC and NO_x and on the performance parameters such as BSFC, torque, thermal and combustion efficiencies have been discussed. The relationships between the emissions are also provided.     

The influence of oxygenated fuels on emissions of aldehydes and ketones from a two-stroke spark ignition engine

A spark-ignited two-stroke chainsaw engine was used to study the influence of pure oxygenated fuels on exhaust emissions of carbonyls (aldehydes and ketones) and regulated emissions, i.e. hydrocarbons (HC), carbon monoxide (CO), and nitrogen oxides (NO_x). Three fuels—methanol, methyl tert-butylether (MTBE), and ethyl tert-butylether (ETBE)—were used in the tests, each at three air/fuel ratios (λ) and the generated emissions were compared to those observed in previous tests with ethanol, aliphatic gasoline, and regular gasoline. Use of all four oxygenated fuels (ETBE, ethanol, methanol and MTBE) resulted in substantially higher total carbonyl emissions (11, 11, 8.9 and 7.8 g/kWh, respectively) than use of both aliphatic and regular gasoline (2.1 and 2.6 g/kWh, respectively). Further, up to 44-fold higher levels of specific carbonyls were generated from the oxygenated fuels than from regular gasoline: significant amounts of formaldehyde were produced from all of the oxygenated fuels, but they were especially high from methanol and MTBE; acetaldehyde was formed in high amounts from ethanol and ETBE; while acetone and methacrolein were formed from both MTBE and ETBE. In addition, increases in λ increased exhaust emissions of formaldehyde, acetaldehyde, acetone, and methacrolein in cases where these were the main carbonyls formed. Increasing λ also variously increased, reduced or had no significant effect on emissions of other measured carbonyls. Lower amounts of CO and NO_x emissions were formed from all oxygenates (especially methanol) than from regular gasoline.     

A20 Overexpression Inhibits Lipopolysaccharide-Induced NF-κB Activation,TRAF6 and CD40 Expression in Rat Peritoneal Mesothelial Cells

Zinc finger protein A20 is a key negative regulator of inflammation. However, whether A20 may affect inflammation during peritoneal dialysis (PD)-associated peritonitis is still unclear. This study was aimed to investigate the effect of A20 overexpression on lipopolysaccharide (LPS)-induced inflammatory response in rat peritoneal mesothelial cells (RPMCs). Isolated and cultured RPMCs in vitro. Plasmid pGEM-T easy-A20 was transfected into RPMCs by Lipofectamine™2000. The protein expression of A20, phospho-IκBα, IκBα, TNF receptor-associated factor (TRAF) 6 and CD40 were analyzed by Western blot. The mRNA expression of TRAF6, CD40, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were determined by real time-PCR. NF-κB p65 DNA binding activity, IL-6 and TNF-α levels in cells culture supernatant were determined by ELISA. Our results revealed that RPMCs overexpression of A20 lead to significant decrease of LPS-induced IκBα phosphorylation and NF-κB DNA binding activity (all p < 0.01). In addition, A20 also attenuated the expression of TRAF6, CD40, IL-6 and TNF-α as well as levels of IL-6 and TNF-α in cells culture supernatant (all p < 0.05). However, A20 only partly inhibited CD40 expression. Our study indicated that A20 overexpression may depress the inflammatory response induced by LPS in cultured RPMCs through negatively regulated the relevant function of adaptors in LPS signaling pathway.     

Comparison of Osteogenic Potentials of Dental Pulp and Bone Marrow Mesenchymal Stem Cells Using the New Cell Transplantation Platform,CellSaic, in a Rat Congenital Cleft-Jaw Model

Scaffolds stimulate cell proliferation and differentiation and play major roles in providing growth and nutrition factors in the repair of bone defects. We used the recombinant peptide Cellnest™ to prepare the three-dimensional stem cell complex, CellSaic, and evaluated whether CellSaic containing rat dental pulp stem cells (rDPSCs) was better than that containing rat bone marrow stem cells (rBMSCs). rDPSC-CellSaic or rBMSC-CellSaic, cultured with or without osteogenic induction medium, formed the experimental and control groups, respectively. Osteoblast differentiation was evaluated in vitro and transplanted into a rat model with a congenital jaw fracture. Specimens were collected and evaluated by microradiology and histological analysis. In the experimental group, the amount of calcium deposits, expression levels of bone-related genes (RUNX2, ALP, BSP, and COL1), and volume of mineralized tissue, were significantly higher than those in the control group (p < 0.05). Both differentiated and undifferentiated rDPSC-CellSaic and only the differentiated rBMSC-CellSaic could induce the formation of new bone tissue. Overall, rBMSC-CellSaic and rDPSC-CellSaic made with Cellnest™ as a scaffold, provide excellent support for promoting bone regeneration in rat mandibular congenital defects. Additionally, rDPSC-CellSaic seems a better source for craniofacial bone defect repair than rBMSC-CellSaic, suggesting the possibility of using DPSCs in bone tissue regenerative therapy.     

Investigation on combustion and emissions of DME/gasoline mixtures in a spark-ignition engine

Dimethyl ether (DME) has a lot of good properties and is thought to be one of the best alternative fuels for IC engines in the future. In order to improve the efficiency, combustion stability and emissions performance of a spark-ignited (SI) gasoline engine at stoichiometric condition, an experimental study aiming at improving engine performance through DME addition was carried out on a four-cylinder SI engine. The engine was modified to be fueled with the mixture of gasoline and DME which were injected into the engine intake ports simultaneously. A hybrid electronic control unit (HECU) was dedicatedly developed to control the injection timings and durations of gasoline and DME. The spark timing was adjusted to reach the maximum brake torque (MBT) without knocking. Various DME fractions were selected to investigate the effect of DME addition on engine performance, thermal efficiency, combustion characteristics, cyclic variation and emissions under stoichiometric conditions. The experimental results showed that thermal efficiency, NOx and HC emissions are improved with the increase of DME addition level. The combustion performance was improved when DME addition fraction was less than 10%. CO emission first decreased and then increased with the increase of DME enrichment level at stoichiometric condition.     

Combustion performance of bio-ethanol at various blend ratios in a gasoline direct injection engine

Bio-ethanol has the potential to be used as an alternative to petroleum gasoline for the purpose of reducing the total CO₂ emissions from internal combustion engines and this paper is devoted to the investigation of using different blending-ratios of bio-ethanol/gasoline with respect to spark timing and injection strategies. The experimental work has been carried out on a direct injection spark ignition engine at a part load and speed condition. It is shown that the benefits of adding ethanol into gasoline are reduced engine-out emissions and increased efficiency, and the impact changes with the blend ratio following a certain pattern. These benefits are attributed to the fact that the addition of ethanol modifies the evaporation properties of the fuel blend which increases the vapour pressure for low blends and reduces the heavy fractions for high blends. This is furthermore coupled with the presence of oxygen within the ethanol fuel molecule and the contribution of its faster flame speed, leading to enhanced combustion initiation and stability and improved engine efficiency.     

Prenylated Flavonoids from Cudrania tricuspidata Suppress Lipopolysaccharide-Induced Neuroinflammatory Activities in BV2 Microglial Cells

In Korea and China, Cudrania tricuspidata Bureau (Moraceae) is an important traditional medicinal plant used to treat lumbago, hemoptysis, and contusions. The C. tricuspidata methanol extract suppressed both production of NO and PGE₂ in BV2 microglial cells. Cudraflavanone D (1), isolated from this extract, remarkably suppressed the protein expression of inducible NO synthase and cyclooxygenase-2, and decreased the levels of NO and PGE₂ in BV2 microglial cells exposed to lipopolysaccharide. Cudraflavanone D (1) also decreased IL-6, TNF-α, IL-12, and IL-1β production, blocked nuclear translocation of NF-κB heterodimers (p50 and p65) by interrupting the degradation and phosphorylation of inhibitor of IκB-α, and inhibited NF-κB binding. In addition, cudraflavanone D (1) suppressed the phosphorylation of c-Jun N-terminal kinase (JNK) and p38 MAPK pathways. This study indicated that cudraflavanone D (1) can be a potential drug candidate for the cure of neuroinflammation.     

Measurement of number and size distribution of particles emitted from a mid-sized transportation multipoint port fuel injection gasoline engine

This study was carried out to characterize the engine-exhaust particulate emissions from a typical multipoint port fuel injection gasoline engine used in transportation sector. Though gasoline engine showed no visible tail pipe emissions yet its particle concentrations were comparable to mineral diesel, particularly at high engine loads. Average sizes of particles emitted in gasoline exhaust are found to be way smaller than particles emitted in diesel exhaust under similar operating conditions. The peak particle concentrations for mineral diesel never go below 40 nm size however for gasoline engine, it was as low as 20 nm for most engine operating conditions. Within a very limited operating range, gasoline engine performance was superior to its diesel counterparts in terms of particulate size and number distribution however it deteriorates very quickly as soon as the fuel-air mixture becomes closer to stoichiometric ratio, typically under high engine load and speed conditions.     

Effect of hydrogen addition on lean burn performance of a spark-ignited gasoline engine at 800 rpm and low loads

To reduce the fuel consumption and emissions of spark-ignited (SI) engines, hydrogen enrichment was used to improve the performance of a lean burn SI engine operating at low speed and load conditions. A hydrogen port-injection system was mounted on the intake manifolds to introduce hydrogen into the intake ports sequentially while keeping the original gasoline injection system unchanged. A hybrid electronic control unit (HECU) was adopted to control injection timings and durations of gasoline and hydrogen, accomplishing four excess air ratios of 1.00, 1.18, 1.43 and 1.67 and three hydrogen volume fractions in the intake of 3%, 5%, 8%. The experimental results showed that engine brake thermal efficiency and torque output were increased, combustion durations were shortened, cyclic variation and HC emissions were reduced, but NO_x emissions were increased with the increase of hydrogen addition. CO emission was also reduced under lean conditions with hydrogen enrichment.     

CK2 Down-Regulation Increases the Expression of Senescence-Associated Secretory Phenotype Factors through NF-κB Activation

Senescent cells secrete pro-inflammatory factors, and a hallmark feature of senescence is senescence-associated secretory phenotype (SASP). The aim of this study is to investigate the protein kinase CK2 (CK2) effects on SASP factors expression in cellular senescence and organism aging. Here CK2 down-regulation induced the expression of SASP factors, including interleukin (IL)-1β, IL-6, and matrix metalloproteinase (MMP) 3, through the activation of nuclear factor-κB (NF-κB) signaling in MCF-7 and HCT116 cells. CK2 down-regulation-mediated SIRT1 inactivation promoted the degradation of inhibitors of NF-κB (IκB) by activating the AKT-IκB kinase (IKK) axis and increased the acetylation of lysine 310 on RelA/p65, an important site for the activity of NF-κB. kin-10 (the ortholog of CK2β) knockdown increased zmp-1, -2, and -3 (the orthologs of MMP) expression in nematodes, but AKT inhibitor triciribine and SIRT activator resveratrol significantly abrogated the increased expression of these genes. Finally, antisense inhibitors of miR-186, miR-216b, miR-337-3p, and miR-760 suppressed CK2α down-regulation, activation of the AKT-IKK-NF-κB axis, RelA/p65 acetylation, and expression of SASP genes in cells treated with lipopolysaccharide. Therefore, this study indicated that CK2 down-regulation induces the expression of SASP factors through NF-κB activation, which is mediated by both activation of the SIRT1-AKT-IKK axis and RelA/p65 acetylation, suggesting that the mixture of the four miRNA inhibitors can be used as anti-inflammatory agents.     

25(OH)D Is Effective to Repress Human Cholangiocarcinoma Cell Growth through the Conversion of 25(OH)D to 1α,25(OH)2D3

Cholangiocarcinoma (CCA) is a devastating disease without effective treatments. 1α,25(OH)₂D₃, the active form of Vitamin D, has emerged as a new anti-cancer regimen. However, the side effect of hypercalcemia impedes its systemic administration. 25(OH)D is biologically inert and needs hydroxylation by CYP27B1 to form 1α,25(OH)₂D₃, which is originally believed to only take place in kidneys. Recently, the extra-renal expression of CYP27B1 has been identified and in vitro conversion of 25(OH)D to 1α,25(OH)₂D₃ has been found in some cancer cells with CYP27B1 expression. In this study, CYP27B1 expression was demonstrated in CCA cells and human CCA specimens. 25(OH)D effectively represses SNU308 cells growth, which was strengthened or attenuated as CYP27B1 overexpression or knockdown. Lipocalcin-2 (LCN2) was also found to be repressed by 25(OH)D. After treatment with 800 ng/mL 25(OH)D, the intracellular 1α,25(OH)₂D₃ concentration was higher in SNU308 cells with CYP27B1 overexpression than wild type SNU308 cells. In a xenograft animal experiment, 25(OH)D, at a dose of 6 μg/kg or 20 μg/kg, significantly inhibited SNU308 cells’ growth without inducing obvious side effects. Collectively, our results indicated that SNU308 cells were able to convert 25(OH)D to 1α,25(OH)₂D₃ and 25(OH)D CYP27B1 gene therapy could be deemed as a promising therapeutic direction for CCA.     

Alzheimer’s Disease Associated Presenilin 1 and 2 Genes Dysregulation in Neonatal Lymphocytes Following Perinatal Asphyxia

Perinatal asphyxia is mainly a brain disease leading to the development of neurodegeneration, in which a number of peripheral lesions have been identified; however, little is known about the expression of key genes involved in amyloid production by peripheral cells, such as lymphocytes, during the development of hypoxic-ischemic encephalopathy. We analyzed the gene expression of the amyloid protein precursor, β-secretase, presenilin 1 and 2 and hypoxia-inducible factor 1-α by RT-PCR in the lymphocytes of post-asphyxia and control neonates. In all examined periods after asphyxia, decreased expression of the genes of the amyloid protein precursor, β-secretase and hypoxia-inducible factor 1-α was noted in lymphocytes. Conversely, expression of presenilin 1 and 2 genes decreased on days 1–7 and 8–14 but increased after survival for more than 15 days. We believe that the expression of presenilin genes in lymphocytes could be a potential biomarker to determine the severity of the post-asphyxia neurodegeneration or to identify the underlying factors for brain neurodegeneration and get information about the time they occurred. This appears to be the first worldwide data on the role of the presenilin 1 and 2 genes associated with Alzheimer’s disease in the dysregulation of neonatal lymphocytes after perinatal asphyxia.     

Geranylgeraniol Inhibits Lipopolysaccharide-Induced Inflammation in Mouse-Derived MG6 Microglial Cells via NF-κB Signaling Modulation

Persistent inflammatory reactions in microglial cells are strongly associated with neurodegenerative pathogenesis. Additionally, geranylgeraniol (GGOH), a plant-derived isoprenoid, has been found to improve inflammatory conditions in several animal models. It has also been observed that its chemical structure is similar to that of the side chain of menaquinone-4, which is a vitamin K₂ sub-type that suppresses inflammation in mouse-derived microglial cells. In this study, we investigated whether GGOH has a similar anti-inflammatory effect in activated microglial cells. Particularly, mouse-derived MG6 cells pre-treated with GGOH were exposed to lipopolysaccharide (LPS). Thereafter, the mRNA levels of pro-inflammatory cytokines were determined via qRT-PCR, while protein expression levels, especially the expression of NF-κB signaling cascade-related proteins, were determined via Western blot analysis. The distribution of NF-κB p65 protein was also analyzed via fluorescence microscopy. Thus, it was observed that GGOH dose-dependently suppressed the LPS-induced increase in the mRNA levels of Il-1β, Tnf-α, Il-6, and Cox-2. Furthermore, GGOH inhibited the phosphorylation of TAK1, IKKα/β, and NF-κB p65 proteins as well as NF-κB nuclear translocation induced by LPS while maintaining IκBα expression. We showed that GGOH, similar to menaquinone-4, could alleviate LPS-induced microglial inflammation by targeting the NF-kB signaling pathway.