Aberrant Active cis-Regulatory Elements Associated with Downregulation of RET Finger Protein Overcome Chemoresistance in Glioblastoma

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Wnt3a upregulates transforming growth factor-β-stimulated VEGF synthesis in osteoblasts

It is recognized that Wnt3a affects bone metabolism via the canonical Wnt/β-catenin signalling pathway. We have previously shown that transforming growth factor-β (TGF-β) stimulates the synthesis of vascular endothelial growth factor (VEGF) via p44/p42 mitogen-activated protein (MAP) kinase, stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK) and p38 MAP kinase in osteoblast-like MC3T3-E1 cells. In the present study, we investigated the effect of Wnt3a on TGF-β-stimulated VEGF synthesis in these cells. Wnt3a, which alone had little effect on the VEGF levels, significantly enhanced the TGF-β-stimulated VEGF release. Lithium chloride and SB216763, inhibitors of glycogen synthase kinase 3β, markedly amplified the TGF-β-stimulated VEGF release. Wnt3a failed to affect the TGF-β-induced phosphorylation of Smad2, p44/p42 MAP kinase, p38 MAP kinase or SAPK/JNK. Wnt3a and lithium chloride strengthened the VEGF mRNA expression induced by TGF-β. These results strongly suggest that Wnt3a upregulates VEGF synthesis stimulated by TGF-β via activation of the canonical pathway in osteoblasts.     

Synthesis and biological evaluation of thielocin B1 analogues as protein-protein interaction inhibitors of PAC3 homodimer

The synthesis and biological evaluation of thielocin B1 analogues have been demonstrated. Fourteen analogues modified in the central core and terminal carboxylic acid moiety were concisely synthesized by simple esterification or etherification reaction. The evaluation of synthetic analogues as inhibitors of proteasome assembling chaperone (PAC) complexes (the PAC3 homodimer and PAC1/PAC2) revealed that the natural product-like bending structure and terminal carboxylic acid groups were crucial for its biological activity. Moreover, SAR and in silico docking studies indicated that all methyl groups on the diphenyl ether moiety of thielocin B1 contribute to the potent and selective inhibition of the PAC3 homodimer via hydrophobic interactions.     

Day workers suffering from a wider range of sleep problems are more likely to experience suicidality

Both a higher suicide rate and widespread sleep problems are serious health concerns in Japan when compared with those of other countries. We investigated the relationship between suicidal ideation and sleep problems in Japanese day workers using the 3-dimensional sleep scale (3DSS), which measures three sleep elements (phase, quality, and quantity). Data from 635 Japanese day workers (461 mens and 174 womens) were included. The 3DSS was used to assess participants’ sleep condition. Participants were classified into eight sleep types based on scores of phase, quality, and quantity: All Good Sleep, Owl (poor phase), Inefficient (poor quality), Short (poor quantity), Owl + Inefficient (poor phase and quality), Owl + Short (poor phase and quantity), Inefficient + Short (poor quality and quantity), and All Poor Sleep. We assessed participants’ suicidal ideation using question 19 of the self-rating depression scale (SDS); 119 cases (18.7 %) had ratings of 2–4 for this question and were considered to have suicidal ideation. The higher the number of sleep problems, the higher the risk of suicidal ideation compared to sleep types not indicative of problems. All Poor Sleep had the highest risk of the eight sleep types. Individuals with Owl + Short, Inefficient + Short, or All Poor Sleep had a significant risk of suicidal ideation even after adjusting for hopelessness and nightmares. Our findings suggested that sleep problems assessed by the 3DSS were related to suicidal ideation. Analysis of various aspects of sleep could be helpful for suicide prevention.

     

Phenomenological description of active transport of salt and water

The phenomenological definition of active transport by Kedem and the methods of Kedem and Katchalsky have been used to obtain practical equations describing active transport in the single salt and bi-ionic systems. Procedures were devised to evaluate the required set of 10 coefficients for the single salt case and 15 for the bi-ionic. Three of these coefficients are unusual. They express the effects of active transport, i.e. of entrainment between metabolism and the conventional transport flows: active salt transport coefficient, a volume pump coefficient, and an electrogenicity coefficient. In the bi-ionic case a new passive coefficient, lambda, was used to express the linkage between the fluxes of the two salts. However, if primary active transport involves only one ion, for example in the bi-ionic case, 12 coefficients suffice and certain relations can be predicted between the practical coefficients. Particular types of primary active transport could be identified by this means. The relation of active transport to membrane electrogenesis was also examined and the flux ratio equation was rederived in terms of the practical coefficients. Applications to specific parallel and series membrane systems have been analyzed.     

Effects of basolateral ouabain, amphotericin B, cyanide and potassium on amiloride noise during voltage clamp of Rana pipiens skin support sodium-amiloride competition

In a previous study, the amiloride-induced corner frequency (fc) was found to decrease as apical sodium was increased. This effect was small or absent when the basolateral surface was exposed to high potassium. It has been suggested that the apical sodium effect may be indirect, due either to increased intracellular [Na+] which repelled amiloride or to an increased potential at the apical surface which reduced amiloride affinity. High basolateral K+ might then suppress the sodium effect either by preventing intracellular [Na+] from increasing or by allowing a better clamp of the apical membrane potential by reducing basolateral membrane resistance and potential. We checked the effects of basolateral [K+], of cyanide and of ouabain at concentrations known to increase intracellular [Na+]. We found only negligible effects on fc. In addition, amphotericin B added to the basolateral bathing solution either in 115 mM Na+ or in 120 mM K+ had no significant effect on fc. We found that relatively wide variation in clamp potential under all conditions, even with active transport severely inhibited, left fc virtually constant. Since the amiloride kinetics were independent of clamp potential, we were able to measure paracellular and transcellular conductances separately by examining the voltage dependence of clamp current (linear) and amiloride noise power (quadratic). This made possible estimation of channel density and single-channel current.     

Analysis of stability and catalytic properties of two tryptophanases from a thermophile.

Two tryptophanases, Tna1 and Tna2, both of which were cloned from the thermophile Symbiobacterium thermophilum, differ in their enzymatic properties, such as thermal stability, catalytic efficiency and activation energy of catalysis, despite the great similarity (92%) in their amino acid sequences. Chimeric tryptophanases were constructed by recombination of the two genes to try to elucidate the molecular basis for the difference. The stability of each chimeric enzyme was roughly proportional to the content of amino acid residues from Tna1. Three regions, tentatively named regions 2, 4 and 5, which contained the amino acid residues 70-129, 192-298 and 299-453, respectively, were especially important for the increase in thermal stability. Site-directed mutagenesis revealed that V104 in region 2 and Y198 in region 4 of Tna1 were involved in the increase in thermal stability of Tna1. Amino acid residues contributing to the higher catalytic efficiency of Tna1 were similarly analyzed, using the chimeric tryptophanases, and found to be located in region 5. Site-directed mutagenesis revealed that I383 and G395 in Tna1, which were presumably located close to the putative active center, played an active role in the increase of catalytic efficiency of Tna1. The activation energy of catalysis was proportional to the content of amino acid residues from Tna2, suggesting the amino acid residues responsible for the difference were dispersed over the whole molecule.     

Acetylation regulates subcellular localization of eukaryotic translation initiation factor 5A (eIF5A)

Eukaryotic translation initiation factor 5A (eIF5A) is a protein subject to hypusination, which is essential for its function. eIF5A is also acetylated, but the role of that modification is unknown. Here, we report that acetylation regulates the subcellular localization of eIF5A. We identified PCAF as the major cellular acetyltransferase of eIF5A, and HDAC6 and SIRT2 as its major deacetylases. Inhibition of the deacetylases or impaired hypusination increased acetylation of eIF5A, leading to nuclear accumulation. As eIF5A is constitutively hypusinated under physiological conditions, we suggest that reversible acetylation plays a major role in controlling the subcellular localization of eIF5A.