Escherichia coli, which harbored the ars operon from a plasmid pMH12 of Klebsiella oxytoca D12, showed filamentation due to the expression of ars genes in the presence of arsenite. The continued DNA replication in the absence of cell division was revealed, since nucleoids abound with DAPI appeared to be arranged in chains. In contrast to overexpression of arsA, its frame-shift mutant and knock-out mutant lost filamentation in the presence of arsenite, which suggested that ars-induced division block was dependent on expression of arsA. ArsA-induced division inhibition was not a consequence of an inhibition of DNA replication, and the inability of arsenite to induce an SOS response indicated that arsA-mediated division inhibition was dependent on the expression of the gene product encoded by the minB operon. ArsA is a peripheral membrane protein with an ATP-binding domain, which is homologous to MinD that requires ATP-dependent efflux. These results suggested that ArsA could possibly recruit MinC to the membrane and modulate cytoplasmic FtsZ to block assembly at the middle of the cell.
Endotoxic shock causes death in humans and animals via extreme hypoperfusion of peripheral organs. A massive production of nitric oxide (NO) both from the endothelical cells and smooth muscle cells has been proposed as a possible mechanism in this process. Since NO attenuated the contractility to vasoconstricting agents such as norepinephrine (NE) by directly acting on the smooth muscle cells, this mechanism was considered mainly as a postsynaptic mechanism. In this research it was investigated whether NO, thus released, also participates in the presynaptic events for the regulation of vascular tone in endotoxic shock. The role of NO was studied by adding NO donors or NO synthase inhibitor $N^\omega $methyl-L-arginine (NMA) in stimulated sympathetic nerves of the mesenteric vascular bed and the Langendorff heart of rats. Sodium nitroprusside (SNP), an NO donor, reduced the pressor responses of isolated mesenteric artery either to electrical stimulation or exogenously administered phenylephrine (PE). In this mesentery, although neither agent influenced NE release, in the presence of the adrenergic $\alpha_2$-receptor antagonist yohimbine, elecrical stimulation-evoked NE release was augumented by SNP. In the heart SNP facilitated the NE release induced by electrical stimulation, while NMA had no effect. From these results it is proposed that there exists a local reflex phenomenon in the junction between the sympathetic nerve terminals and the smooth muscle of resistance blood vessels; by which sympathetic responses are reduced by NO at the postjunctional level while NO facilitates NE release contributing to augumentation of sympathetic tone. All these facts suggest that NO produced during endotoxic shock has dual effects: whereas NO blunts the vasoconstrictive activity of NE at the postsynaptic level, NO presynaptically facilitates the release of NE from sympathetic nerve terminals.
In the present study, buckling analysis of sandwich composite (carbon nanotube reinforced composite and fiber reinforced composite) Euler-Bernoulli beam in two configurations (core and layers material), three laminates (combination of different angles) and two models (relative thickness of core according to peripheral layers) using differential quadrature method (DQM) is studied. Also, the effects of porosity coefficient and different types of porosity distribution on critical buckling load are discussed. Using sandwich beam, it shows a considerable enhancement in the critical buckling load when compared to ordinary composite. Actually, resistance against buckling in sandwich beam is between two to four times more. It is also showed the critical buckling loads of laminate 1 and 3 are significantly larger than the results of laminate 2. When Configuration 2 is used, the critical buckling load rises about 3 percent in laminate 1 and 3 compared to the results of configuration 1. The amount of enhancement for laminate 3 is about 17 percent. It is also demonstrated that the influence of the core height (thickness) in the case of lower carbon volume fractions is ignorable. Even though, when volume fraction of fiber increases, differences grow smoothly. It should be noticed the amount of decline has inverse relationship with the beam aspect ratio. Among three porosity patterns investigated, beam with the distribution of porosity Type 2 (downward parabolic) has the maximum critical buckling load. At the end, the first three modes of buckling will be demonstrated to investigate the effect of spring constants.
Fu, Zhicheng;Yun, So Yoon;Won, Jong Hoon;Back, Moon Jung;Jang, Ji Min;Ha, Hae Chan;Lee, Hae Kyung;Shin, In Chul;Kim, Ju Yeun;Kim, Hee Soo;Kim, Dae Kyong
Biomolecules & Therapeutics
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제27권2호
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pp.193-200
/
2019
Ceramide metabolism is known to be an essential etiology for various diseases, such as atopic dermatitis and Gaucher disease. Glucosylceramide synthase (GCS) is a key enzyme for the synthesis of glucosylceramide (GlcCer), which is a main ceramide metabolism pathway in mammalian cells. In this article, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine GCS activity using synthetic non-natural sphingolipid C8-ceramide as a substrate. The reaction products, C8-GlcCer for GCS, could be separated on a C18 column by reverse-phase high-performance liquid chromatography (HPLC). Quantification was conducted using the multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z $588.6{\rightarrow}264.4$ for C8-GlcCer at positive ionization mode. The calibration curve was established over the range of 0.625-160 ng/mL, and the correlation coefficient was larger than 0.999. This method was successfully applied to detect GCS in the human hepatocellular carcinoma cell line (HepG2 cells) and mouse peripheral blood mononuclear cells. We also evaluated the inhibition degree of a known GCS inhibitor 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) on GCS enzymatic activity and proved that this method could be successfully applied to GCS inhibitor screening of preventive and therapeutic drugs for ceramide metabolism diseases, such as atopic dermatitis and Gaucher disease.
Internal ring beams are primary components of new ring-stiffened cooling towers. In this study, numerical simulation of the internal flow field of a cooling tower with three ring beams under wind-thermal coupling effect is performed. The studied cooling tower is a 220-m super-large hyperbolic indirect natural draft cooling tower that is under construction in China and will be the World's highest cooling tower, the influence of peripheral radiators in operating cooling tower is also considered. Based on the simulation, the three-dimensional effect and distribution pattern of the wind loads on inner surface of the cooling tower is summarized, the average wind pressure distributions on the inner surface before and after the addition of the ring beams are analyzed, and the influence pattern of ring beams on the internal pressure coefficient value is derived. The action mechanisms behind the air flows inside the tower are compared. In addition, the effects of internal ring beams on temperature field characteristics, turbulence kinetic energy distribution, and wind resistance are analyzed. Finally, the internal pressure coefficients are suggested for ring-stiffened cooling towers under wind-thermal coupling effect. The study shows that the influence of internal stiffening ring beams on the internal pressure and flow of cooling towers should not be ignored, and the wind-thermal coupling effect should also be considered in the numerical simulation of cooling tower flow fields. The primary conclusions presented in this paper offer references for determining the internal suction of such ring-stiffened cooling towers.
Objective: Approximately 30% of preeclamptic pregnancies exhibit abnormal liver function tests. We assessed liver injury-associated enzyme levels and circulating transforming growth factor beta (TGF-β) levels in an arginine vasopressin (AVP)-induced pregnant Sprague-Dawley rat model. Methods: Pregnant and non-pregnant Sprague-Dawley rats (n=24) received AVP (150 ng/hr) subcutaneously via mini-osmotic pumps for 18 days. Blood pressure was measured, urine samples were collected, and all animals were euthanized via isoflurane. Blood was collected to measure circulating levels of TGF-β1-3 isomers and liver injury enzymes in pregnant AVP (PAVP), pregnant saline (PS), non-pregnant AVP (NAVP), and non-pregnant saline (NS) rats. Results: The PAVP group showed significantly higher systolic and diastolic blood pressure than both saline-treated groups. The weight per pup was significantly lower in the AVP-treated group than in the saline group (p<0.05). Circulating TGF-β1-3 isomer levels were significantly higher in the PAVP rats than in the NS rats. However, similar TGF-β1 and TGF-β3 levels were noted in the PS and PAVP rats, while TGF-β2 levels were significantly higher in the PAVP rats. Circulating liver-type arginase-1 and 5'-nucleotidase levels were higher in the PAVP rats than in the saline group. Conclusion: This is the first study to demonstrate higher levels of TGF-β2, arginase, and 5'-nucleotidase activity in PAVP than in PS rats. AVP may cause vasoconstriction and increase peripheral resistance and blood pressure, thereby elevating TGF-β and inducing the preeclampsia-associated inflammatory response. Future studies should explore the mechanisms through which AVP dysregulates liver injury enzymes and TGF-β in pregnant rats.
Background: We have reported that internal deletions in the nef, gag, and pol genes in HIV-1-infected patients are induced in those treated with Korean Red Ginseng (KRG). KRG delays the development of resistance mutations to antiretroviral drugs. Methods: The vif-vpr genes over 26 years in 20 hemophiliacs infected with HIV-1 from a single source were sequenced to investigate whether vif-vpr genes were affected by KRG and KRG plus highly active antiretroviral therapy (ART) (hereafter called GCT) and compared the results with our previous data. Results: A significantly higher number of in-frame small deletions were found in the vif-vpr genes of KRG-treated patients than at the baseline, in control patients, and in ART-alone patients (p < 0.001). These were significantly reduced in GCT patients (p < 0.05). In contrast, sequences harboring a premature stop codon (SC) were more significant in GCT patients (10.1%) than in KRG-alone patients, control (p < 0.01), and ART-alone patients (p = 0.078 for peripheral blood mononuclear cells). The proportion of SC in Vpr was similar to that in Vif, whereas the proportion of sequences revealing SC in the env-nef genes was significantly lower than that in the pol-vif-vpr genes (p < 0.01). The genetic distance was 1.8 times higher in the sequences harboring SC than in the sequences without SC (p < 0.001). Q135P in the vif gene is significantly associated with rapid progression to AIDS (p < 0.01). Conclusion: Our data show that KRG might induce sD in the vif-vpr genes and that vif-vpr genes are similarly affected by lethal mutations.
In the late nineteenth century, British colonial activities became aggressive and annexationist in the tropics, including the Southeast Asian Archipelago, which reflected the historical circumstances of both increasing resistance from the indigenous and severe competition among European powers. Interestingly, the change in English colonial policy toward an annexationist or imperialist vision adopted the motto of a civilizing mission, which was founded on the anthropological assumption that the white English were civilized, while the non-white indigenous were savage. The assumption developed into colonial discourse through systematic gathering of anthropological knowledge about the peripheries of the Empire. The knowledge system was flawed, which stressed the differences of the peripheral populations from the English and served as an inverted discourse on the Imperial Self rather than the description of the Other. Furthermore, the natives were heterogeneous, which rendered indistinct the racial and cultural differences between the English and the natives. Still, the aboriginals called Malays, who were comprised of many ethnic subgroups, needed to be deemed savage or inferior by the English in order to justify the English civilizing work or imperial ambition. Put differently, the representation of the English as civilized necessitated the (mis)representation of the natives as savage. In this context, Clifford's works contribute to systematic misrepresentation of the Malays, on which colonial discourse is founded, though not without self-contradiction. On the other hand, Conrad's novels that are set in the Malay Archipelago resort to a strategic misrepresentation that reveals the relativity of the discourse. Exploring the dilemma of denationalization to various degrees, Conrad's Malay texts problematize the (mis)representation of the indigenous as inferior, which is the basis of English claim to superiority.
Hydroxychavicol, a primary active phenolic compound of betel leaves, previously inhibited bone loss in vivo by stimulating osteogenesis. However, the effect of hydroxychavicol on bone remodeling induced by osteoclasts is unknown. In this study, the anti-osteoclastogenic effects of hydroxychavicol and its mechanism were investigated in receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclasts. Hydroxychavicol reduced the number of tartrate resistance acid phosphatase (TRAP)-positive multinucleated, F-actin ring formation and bone-resorbing activity of osteoclasts differentiated from RAW264.7 cells in a concentration-dependent manner. Furthermore, hydroxychavicol decreased the expression of osteoclast-specific genes, including cathepsin K, MMP-9, and dendritic cell-specific transmembrane protein (DC-STAMP). For mechanistic studies, hydroxychavicol suppressed RANKL-induced expression of major transcription factors, including the nuclear factor of activated T-cells 1 (NFATc1), c-Fos, and c-Jun. At the early stage of osteoclast differentiation, hydroxychavicol blocked the phosphorylation of NF-κB subunits (p65 and Iκβα). This blockade led to the decrease of nuclear translocation of p65 induced by RANKL. In addition, the anti-osteoclastogenic effect of hydroxychavicol was confirmed by the inhibition of TRAP-positive multinucleated differentiation from human peripheral mononuclear cells (PBMCs). In conclusion, hydroxychavicol inhibits osteoclastogenesis by abrogating RANKL-induced NFATc1 expression by suppressing the NF-κB signaling pathway in vitro.
Kim, Kyung Min;Kim, Hyo Sup;Yoon, Ji Hong;Lee, Eun-Jung;Yum, Sook Kyung;Moon, Cheong-Jun;Youn, Young-Ah;Kwun, Yoo Jin;Lee, Jae Young;Sung, In Kyung
Neonatal Medicine
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제25권2호
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pp.78-84
/
2018
Purpose: To investigate the hemodynamic risk factors for necrotizing enterocolitis (NEC), we analyzed the characteristics of descending aorta (DA) blood flow in preterm neonates, who later developed NEC. Methods: This was an observational case-control study on 53 preterm neonates at a tertiary referral center. Clinical and echocardiographic data were collected from 23 preterm neonates with NEC (NEC group), and compared with those of 30 preterm neonates without NEC (control group). Echocardiography was done at a median (interquartile range) of 5 (3-9) days after birth and 2 (1-2.5) days before the diagnosis of NEC. Results: Basic clinical characteristics including gestational age, birth weight, Apgar score, breast feeding status, use of umbilical catheters, and mode of invasive ventilator care were similar between the groups. Compared with the control group, the lowest diastolic velocity of DA was significantly decreased, whereas the diastolic reverse flow and the ratio of diastolic reverse to systolic forward flows were significantly increased in the NEC group. In addition, the resistive index (RI) of DA was significantly increased in the NEC group and showed a positive association with the development of NEC. Multivariate logistic regression analysis showed that increasing RI of DA was an independent risk factor for the development of NEC (P=0.008). Conclusion: Significant changes in DA flow characteristics including decreased diastolic velocity and increased diastolic reverse flow along with increased peripheral vascular resistance were observed before the development of NEC in preterm neonates. These findings may help clinicians stratify in advance neonates at a risk of developing NEC and may help improve outcomes in these neonates.
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