PIM kinases facilitate lentiviral evasion from SAMHD1 restriction via Vpx phosphorylation

Lentiviruses have evolved to acquire an auxiliary protein Vpx to counteract the intrinsic host restriction factor SAMHD1. Although Vpx is phosphorylated, it remains unclear whether such phosphorylation indeed regulates its activity toward SAMHD1. Here we identify the PIM family of serine/threonine protein kinases as the factors responsible for the phosphorylation of Vpx and the promotion of Vpx-mediated SAMHD1 counteraction. Integrated proteomics and subsequent functional analysis reveal that PIM family kinases, PIM1 and PIM3, phosphorylate HIV-2 Vpx at Ser13 and stabilize the interaction of Vpx with SAMHD1 thereby promoting ubiquitin-mediated proteolysis of SAMHD1. Inhibition of the PIM kinases promotes the antiviral activity of SAMHD1, ultimately reducing viral replication. Our results highlight a new mode of virus–host cell interaction in which host PIM kinases facilitate promotion of viral infectivity by counteracting the host antiviral system, and suggest a novel therapeutic strategy involving restoration of SAMHD1-mediated antiviral response

Synchronous bilateral pheochromocytomas and paraganglioma with novel germline mutation in MAX: a case report

BackgroundRecent advance of genetic testing has contributed to the diagnosis of hereditary pheochromocytoma and paraganglioma (PPGL). The clinical characteristics of hereditary PPGL are varying among the types of mutational genes. It is still difficult to specify the pathognomonic symptoms in the case of rare genetic mutations. Here, we report the case of synchronous bilateral pheochromocytomas and paraganglioma with novel MYC associated factor X (MAX) gene mutation.Case presentationA 24-year-old female had hyperhidrosis and hypertension. Her urine test showed high normetanephrine and vanillylmandelic acid. Enhanced computed tomography revealed three enhanced masses in right adrenal gland, left adrenal gland, and left renal hilus. She was diagnosed with PPGL. Because 123I-metaiodobenzylguanidine scintigraphy indicated the accumulations in the left adrenal gland mass and the left renal hilus mass and not in the right adrenal gland mass, we performed laparoscopic left adrenalectomy and extirpation of the left renal hilus mass to preserve the right adrenocortical function. However, her symptoms recurred shortly after the operation presumably due to unveiling of the activity of the right pheochromocytoma. Following right adrenalectomy as the second operation, the catecholamine levels declined to normal range. Her genetic testing indicated the novel germline mutation in MAX gene (c.70_73 del AAAC/p.Lys24fs*40).ConclusionsMAX germline mutation is recently identified as a rare cause of hereditary PPGL. The deletion mutation in MAX gene in this patient has never reported before. In the case of bilateral pheochromocytomas, the surgical indication should be decided considering each patient’s genetic background. Due to the possibility for other types of malignant tumors, close follow-up is essential for MAX mutation carriers

岡山県内保育所の園児定員数の推移調査(1)

本資料は,岡山県内に平成11年現存している保育所404園を対象に,設置年,保育所の沿革,措置児童数の推移,措置定員の推移,特別保育事業の開始年度等についての調査を実施した際,明らかになった園児定員数の推移結果を2編にわたってまとめた第1編(市別編)である。本編では岡山県内10市の保育所の昭和23年から平成11年までの園児定員数の全体推移(表1)と公立・私立別の推移(表2)を分けて掲載している

岡山県内保育所の園児定員数の推移調査(2)

本資料は,岡山県内の保育所404園を対象に,設置年,保育所の沿革,措置児童数の推移,措置定員の推移,特別保育事業の開始年度等についての調査を実施した際,明らかになった園児定員数の推移結果を2編にわたってまとめた第2編(郡部別編)である。本編では岡山県内18郡の保育所の昭和23年から平成11年までの園児定員数を公立・私立別にまとめ,第1編にまとめている岡山県10市の園児定員数の推移結果とあわせて比較検討し考察を行っている

Effect of oil droplet size on activation energy for coalescence of oil droplets in an O/W emulsion.

Published online: 12 May 2015The activation energy of a reasonable order of magnitude was estimated for the coalescence of oil droplets in an O/W emulsion by formulating the balance of forces acting on a droplet that crosses over the potential barrier to coalesce with another droplet by the DLVO theory and Stokes' law. An emulsion with smaller oil droplets was shown to be more stable

Production of 5-hydroxymethylfurfural in a eutectic mixture of citric acid and choline chloride and its extractive recovery

Synthesis of 5-hydroxymethylfurfural (HMF) from d-fructose was analyzed from the kinetic and equilibrium points of view in a eutectic mixture of citric acid and choline chloride (CA/CC) or in [Bmim]Cl to achieve an efficient production of HMF in a biphasic reaction system. The partition coefficient of HMF between an organic solvent and the ionic liquid phases was measured using various organic solvents. Among these solvents, ethyl acetate was found to exhibit the best performance for the extractive recovery of HMF. The stability of HMF was also evaluated in CA/CC at 70–120 °C, and HMF was found to be stable below 80 °C. Kinetic analysis of the formation of HMF was performed under the assumption that fructose was converted into HMF through parallel and consecutive reactions. The kinetically calculated results matched well with the experimental ones. The reaction behaviors were different at low and high (over 20% (kg/kg of IL)) initial fructose concentrations. At high initial fructose concentrations, the equilibrium yield of HMF decreased with increasing initial fructose concentration. The addition of ethyl acetate to the CA/CC phase (biphasic reaction) was effective in improving the HMF yield, especially when the fructose concentration was high

Structural implications of the EL(K/Q)(L/C)LD(A/G)DD sequence in the C-terminal cytoplasmic tail for proper targeting of anion exchanger 1 to the plasma membrane

While the C-terminal cytoplasmic tail of anion exchanger 1 (AE1, band 3) has been reported to possess important physiological roles, including one for proper membrane trafficking, its precise characteristics remain unclear. To clarify the overall structural consequences of the conserved sequence EL(K/Q)(L/C)LD(A/G)DD, containing the core binding sequence LDADD for carbonic anhydrase II, in the C-terminal region, we analyzed the membrane expression and turnover of bovine AE1 with a series of truncation and substitution mutations in HEK293 cells. Immunofluorescence microscopy and cell-surface biotinylation demonstrated that truncation mutants missing 18 C-terminal residues targeted the plasma membrane, but the one lacking the conserved region, by truncation of 28 amino acid residues, was retained inside the cells. Substitutions of Ala for Glu901, Leu902, Leu905, and Asp906 in the sequence E901L(K/Q)(L/C)LDADD909 of bovine AE1 or those in the corresponding murine sequence also caused intracellular retention, though these mutants had half-lives comparable to that for wild-type AE1. These data demonstrate that the conserved amino acid residues Glu1, Leu2, Leu5, and Asp6 in the EL(K/Q)(L/C)LD(A/G)DD region have essential structural consequences in stable expression of AE1 at the plasma membrane regardless of the ability in binding to carbonic anhydrase II of this region

Kinetic analysis for the conversion of fructose to 5-hydroxymethylfurfural in 1-butyl-3-methyl imidazolium chloride with lower water contents

Conversion of fructose to 5-hydroxymethylfurfural (HMF) was conducted in an ionic liquid (IL), 1-butyl-3-methylimidazolium chloride ([BMIM]Cl), with low-to-middle (≤ 0.03‒10%) water contents at 100°C with p-toluenesulfonic acid as a catalyst. When fructose was mixed with [BMIM]Cl to start the reaction, fructose dissolved much faster and more rapidly converted to HMF in [BMIM]Cl with a low water content than it did in that with a middle one, suggesting that the conversion rate greatly depended on the said water content. HMF was stable in pure [BMIM]Cl at 100°C, and there were few possibilities for its rehydration and decomposition to byproducts. Based on these results, we proposed an autocatalytic model of the reaction that considers the effects of water; the formation of HMF was certainly affected by the water produced during the conversion. The kinetic analysis based on this model well expressed the formation of HMF in the IL with a low water content