Transcriptional control of occludin expression in vascular endothelia: regulation by Sp3 and YY1

Endothelium differentiates in response to tissue-specific signals; brain endothelium expresses tight junctions and transporters which are absent from other endothelia. The promoter of the tight junction protein occludin exhibited strong activity in a brain endothelial cell line, hCMEC/D3 but was inactive in lung endothelial cells. Expression of occludin in brain endothelium corresponded with binding of Sp3 to a minimal promoter segment close to the transcription-start site. However, in lung endothelium Sp-transcription factors did not bind to this site although they are present in the cell nucleus. In contrast, repression of occludin in lung endothelium was associated with the binding of YY1 to a remote site in the promoter region, which was functionally inactive in brain endothelium. The work identified a group of transcription factors including Sp3 and YY1, which differentially interact with the occludin promoter to induce expression of occludin in brain endothelium and repression in other endothelia. The mechanism controlling occludin expression is similar to that which controls tissue-specific expression of the transferrin receptor in brain endothelium, leading to a scheme for endothelial differentiation, in which activation or repression of tissue-specific proteins is maintained by a set of transcription factors which include Sp3 and YY1

Strategies to increase austenite FCC relative phase stability in high-Mn steels

Several strategies to increase the FCC austenite stability compared to BCC and HCP martensites have been tested and are discussed. The relative stability of the different phases was analyzed by studying the effects of: a) grain size, b) antiferromagnetic ordering of the austenite, c) thermal cycling through the FCC-HCP transition, d) plastic deformation of the austenite and e) combined effects. As a first step, the effect of decreasing the grain size was analyzed in Fe-Mn alloys for Mn contents smaller than 18 wt.%, where BCC and HCP martensites compete in stability. Formation of the BCC phase is inhibited for 15 wt.% and 17 wt.% of Mn for grain sizes smaller than 2 μm. This enabled, for the first time at these compositions, the measurement of the Neel temperature of the austenite using specific heat and magnetic measurements. A comparison of the obtained transition temperatures with accepted models is discussed. The effect of modifying the grain size on the FCC-HCP transition temperatures was also analyzed for 15 wt.% and 17 wt.% Mn contents showing a complete HCP inhibition for grain sizes smaller than 200 nm. A nucleation model for the HCP martensite is considered which includes an additional resistance to the transformation term depending on the austenitic grain size. Additional combined effects on the FCC stabilization are discussed like the interaction between the antiferromagnetic ordering and the introduction of defects by thermal cycling through the martensitic transformation. The analysis can be easily applied to systems with a larger number of components. Results obtained in the Fe-Mn-Cr system are also presented.The authors acknowledge the financial support from ANPCyT (PICT-2017-2198), CONICET (PIP 2015-112-201501-00521), CONICET (PIP 2017e2019 GI 0634), ANPCyT (PICT-2017-4518), and Universidad Nacional de Cuyo (06/C516 and 06/C588)

Expression and localization of claudins-3 and -12 in transformed human brain endothelium

<p>Abstract</p> <p>Background</p> <p>The aim of this study was to characterize the hCMEC/D3 cell line, an <it>in vitro </it>model of the human Blood Brain Barrier (BBB) for the expression of brain endothelial specific claudins-3 and -12.</p> <p>Findings</p> <p>hCMEC/D3 cells express claudins-3 and -12. Claudin-3 is distinctly localized to the TJ whereas claudin -12 is observed in the perinuclear region and completely absent from TJs. We show that the expression of both proteins is lost in cell passage numbers where the BBB properties are no longer fully conserved. Expression and localization of claudin-3 is not modulated by simvastatin shown to improve barrier function <it>in vitro </it>and also recommended for routine hCMEC/D3 culture.</p> <p>Conclusions</p> <p>These results support conservation of claudin-3 and -12 expression in the hCMEC/D3 cell line and make claudin-3 a potential marker for BBB characteristics <it>in vitro</it>.</p

Betanodavirus infection in marine fish aquaculture in Malaysia

Betanodavirus is known to cause mass mortality in many marine aquaculture fish species. In this study, we detected the virus in four different marine aquaculture fish species in Malaysia. These included humpback grouper (Cromileptisaltivelis), brown marbled grouper (Epinephelusfuscoguttatus), Asian seabass (Latescalcarifer) and golden pompano (Trachinoltusblochii). Out of 246 fish specimens analyzed using RT-PCR, 60.98% detected infected by the virus. Histological pathological analysis showed extensive cell vacuolationin the brain and retina tissues of severely infected specimens. However, some of the fish specimens detected positive using RT-PCR did not exhibit cell vacoulation which indicate the carrier state of those specimens. The RT-PCR amplification method developed in this study was shown useful as biosecurity tool in monitoringBetanodavirus infection in aquaculture. Although the origin of Betanodavirus in Malaysia is difficult to ascertained, evidence showed that some infections may have been contributed by the importation of fish fingerlings form neighboring countries. Currently, effective treatment of the viral disease is still impossible hence strict biosecurity measures need to be carried out in order to control the spread of the virus in fish stocks. These can include enforcement of biosecurity check and quarantine of every batch of imported fish, the use of virus-free broodstocks in hatchery, and proper disposal of infected fish stocks. In addition, good aquaculture practices must be carried in aquaculture farms or fish nursery all the time