The integrin-binding defective FGF2 mutants potently suppress FGF2 signalling and angiogenesis.

We recently found that integrin αvβ3 binds to fibroblast growth factor (FGF)-αvβ31 (FGF1), and that the integrin-binding defective FGF1 mutant (Arg-50 to glutamic acid, R50E) is defective in signalling and antagonistic to FGF1 signalling. R50E suppressed angiogenesis and tumour growth, suggesting that R50E has potential as a therapeutic. However, FGF1 is unstable, and we had to express R50E in cancer cells for xenograft study, since injected R50E may rapidly disappear from circulation. We studied if we can develop antagonist of more stable FGF2. FGF2 is widely involved in important biological processes such as stem cell proliferation and angiogenesis. Previous studies found that FGF2 bound to αvβ3 and antagonists to αvβ3 suppressed FGF2-induced angiogenesis. However, it is unclear how FGF2 interacts with integrins. Here, we describe that substituting Lys-119/Arg-120 and Lys-125 residues in the predicted integrin-binding interface of FGF2 to glutamic acid (the K119E/R120E and K125E mutations) effectively reduced integrin binding to FGF2. These FGF2 mutants were defective in signalling functions (ERK1/2 activation and DNA synthesis) in NIH3T3 cells. Notably they suppressed, FGF2 signalling induced by WT FGF2 in endothelial cells, suggesting that the FGF2 mutants are antagonists. The FGF2 mutants effectively suppressed tube formation in vitro, sprouting in aorta ring assays ex vivo and angiogenesis in vivo The positions of amino acids critical for integrin binding are different between FGF1 and FGF2, suggesting that they do not interact with integrins in the same manner. The newly developed FGF2 mutants have potential as anti-angiogenic agents and useful tools for studying the role of integrins in FGF2 signalling

気管支喘息症例における末梢血白血球のロイコトリエンC4産生能とロイコトリエン受容体括抗薬プランルカストの効果に関する検討

The correlation between the efficacy of 4-weeks administration with pranlukast, leukotriene receptor antagonist, and LTs generation by peripheral leukocytes were evaluated in 18 patients with mild-persistent asthma. The efficacy of pranlukast administration was assessed by symptom, morning PEF and pulmonary function. Pranlukast were effective in 12/18(67%) patients. In those patients, LTC4 generation before pranlukast administration was significantly high, compared with that in pranlukast-ineffective patients. LTC4 generation decreased after 4-weeks administration with pranlukast in effective patients. In ineffective patients, however, LTC4 generation increased after 4-weeks administration. LTB4 had shown no significant difference between effective and ineffective patients before administration, and LTB4 decreased after 4-weeks in both groups. Proport ion of peripheral eosinophi Is in effective patients were higher than that in ineffective patients, however not significant. After 4-weeks, proportion of eosinophi Is was decreased in effective patients and increased in ineffective patients. These findings suggest that pranlukast is effective for patients with high LTC4 generation and has the effect to suppress the accumulation of eosinophils in such patients.軽症気管支喘息18例にロイコトリエン受容体桔抗薬プランルカストを4週間投与し，その効果と末梢血白血球からのLTC4，LTB4産生能の関係を検討した．プランルカストの効果は臨床症状，起床時ピークフロー値，肺機能の変化によって判定し，効果群，非効果群の2群に分類した．18例中12例（67％）の症例がプランルカスト投与により，臨床症状の軽減，ピークフロー値の増加，肺機能の改善が認められた．効果群におけるプランルカスト投与前のLTC4値は，非効果群のLTC4値に比較して有意に高値であった．4週間の投与後には 効果群ではLTC4値は減少し，非効果群では増加した．両群のLTB4値はプランルカスト投与前で有意な差は認められず，投与後には両群で減少した．投与前の好酸球分画は，効果群において非効果群に比べ高値であったが，有意な差は認められなかった．4週間の投与後，効果群においては好酸球は減少し，非効果群においては増加した．以上の結果より，プランルカストは末梢血白血球のLTC4産生能が高い症例において効果的であり，好酸球集積を抑制する作用を有すると考えられる

Religion and Politics in Contemporary World

書

気管支喘息患者における白血球ロイコトリエン産生能に対する不飽和脂肪酸食の効果に影響する因子

Dietary supplementation with perilla seed oil, a vegetable oil rich in α -lin- olenic acid, inhibits the generation of leukotrienes(LTs) by leucocytes in patients with bronchial asthma. We examined the factors that affect the suppression of LT generation by leucocytes with perilla seed oil-rich supplementation in patients with asthma, by comparing the clinical features of patients with asthma, whose generation of leukotriene (LT) C4 was suppressed by dietary supplementation with perilla seed oil (n-3 fatty acids) (group A), with those of patients who showed no suppression of LTC4 generation (group B). Group A showed a significant increase in the generation of LTB4 and L TC4 by leucocytes after corn oil-rich supplementation (n-6 fatty acids), and a significant decrease in the generation of LTB4 and LTC4 after perilla seed oil-rich supplementation (n-3 fatty acid). However, this was not observed in group B. The level of serum IgE and peak expiratory flow (PEF) in group A were significantly higher than in group B. Furthermore, the serum levels of LDL-cholesterol, β-lipoprotein and　phospholipid were significantly lower in group A than in group B. These results suggest that the clinical features differ between these two asthmatic populations with respect to suppression of LTB4 and LTC4 generation by n-3 fatty acids in perilla seed oil-rich supplementation.a-リノレン酸の豊富なエゴマ油の食事は気管支喘息患者の白血球ロイコトリエン(LT)産生能を抑制する｡気管支喘息患者の内,エゴマ油食によりLTC4の産生が抑制された群(A群)と抑制されない群(B群)の臨床データを比較することにより,気管支喘息患者の白血球ロイコトリエン産生能に影響する因子を検討した｡A群はコーン油(n-6系脂肪酸)の豊富な食事後,白血LTB4,LTC4の産生能が増加し,エゴマ油(n-3系脂肪酸)の豊富な食事後LTB4,LTC4の産生能が減少した｡これらの変化はB群では認められなかった｡A群のIgE値,ピークフロー(PEF)値はB群に比し,有意に高値であった｡またLDL-コレステロール,β-リポ蛋白,リン脂質はA群ではB群に比し,有意に低値であった｡これらの結果はエゴマ油の豊富な食事のn-3系脂肪酸によるLTB4,LTC4の産生能の抑制に関して2群の気管支喘息患者群間に臨床データの相違があることを示唆している

Efficient derivation and banking of clinical-grade human embryonic stem cell lines in accordance with Japanese regulations

[Introduction] We recently established clinical-grade human embryonic stem cell (hESC) line KthES11 in accordance with current good manufacturing practice standards in Japan. Despite this success, the establishment efficiency was very low at 7.1% in the first period. [Methods] To establish clinical-grade hESC lines, we used xeno-free chemically defined medium StemFit AK03N with the LM-E8 fragments instead of feeder cells. The protocol was then optimized, especially in the early culture phase. [Results] We established five hESC lines (KthES12, KthES13, KthES14, KthES15, and KthES16) with 45.5% efficiency. All five hESC lines showed typical hESC-like morphology, a normal karyotype, pluripotent state, and differentiation potential for all three germ layers. Furthermore, we developed efficient procedures to prepare master cell stocks for clinical-grade hESC lines and an efficient strategy for quality control testing. [Conclusions] Our master cell stocks of hESC lines may contribute to therapeutic applications using human pluripotent stem cells in Japan and other countries

Bafilomycin A1-sensitive pathway is required for the maturation of cystic fibrosis transmembrane conductance regulator

AbstractCystic fibrosis (CF) is the most common lethal genetic disease in Caucasians caused by the trafficking defects of CF transmembrane conductance regulator (CFTR), which is a cAMP-dependent Cl− channel at the plasma membrane. The trafficking pathway of CFTR is thought to be non-conventional because CFTR maturation is inhibited by the dysfunction of syntaxin 13, which is involved in protein recycling via endosomal pathway. In this study, to clarify whether the endosomal trafficking is required for CFTR maturation, we utilized a specific vacuolar H+-ATPase inhibitor, bafilomycin A1 (BafA1), which inhibits the protein trafficking from early endosome. Our data showed that low concentration of BafA1 (50nM) decreased the expression of mature CFTR but induced the accumulation of immature CFTR in the juxta-nuclear region containing an early endosome marker. Pulse-chase analysis showed that BafA1 inhibited the maturation of CFTR, but it slightly stabilized immature CFTR. These results indicate that BafA1-sensitive pathway is required for CFTR maturation and emphasize that endosomal trafficking pathway might be involved in the maturation of CFTR

Characterization of a Novel Bat Adenovirus Isolated from Straw-Colored Fruit Bat (Eidolon helvum).

Bats are important reservoirs for emerging zoonotic viruses. For extensive surveys of potential pathogens in straw-colored fruit bats (Eidolon helvum) in Zambia, a total of 107 spleen samples of E. helvum in 2006 were inoculated onto Vero E6 cells. The cell culture inoculated with one of the samples (ZFB06-106) exhibited remarkable cytopathic changes. Based on the ultrastructural property in negative staining and cross-reactivity in immunofluorescence assays, the virus was suspected to be an adenovirus, and tentatively named E. helvum adenovirus 06-106 (EhAdV 06-106). Analysis of the full-length genome of 30,134 bp, determined by next-generation sequencing, showed the presence of 28 open reading frames. Phylogenetic analyses confirmed that EhAdV 06-106 represented a novel bat adenovirus species in the genus Mastadenovirus. The virus shared similar characteristics of low G + C contents with recently isolated members of species Bat mastadenoviruses E, F and G, from which EhAdV 06-106 diverged by more than 15% based on the distance matrix analysis of DNA polymerase amino acid sequences. According to the taxonomic criteria, we propose the tentative new species name "Bat mastadenovirus H". Because EhAdV 06-106 exhibited a wide in vitro cell tropism, the virus might have a potential risk as an emerging virus through cross-species transmission

Gambling symptoms, behaviors, and cognitive distortions in Japanese university students

Background: This study was conducted to investigate the relationship between symptoms of gambling problems, gambling behaviours, and cognitive distortions among a university student population in Japan ages 20 to 29 years. We aimed to address the gap in knowledge of gambling disorders and treatment for this population. Methods: Data were obtained from 1471 Japanese undergraduate students from 19 universities in Japan. Descriptive statistics and hierarchical multivariate regression analysis were used to investigate whether the factors of gambling cognitive distortions would have predictive effects on gambling disorder symptoms. Results: Results indicated that 5.1% of the participants are classifiable as probable disordered gamblers. The bias of the gambling type to pachinko and pachislot was unique to gamblers in Japan. Of the students sampled, 342 self-reported gambling symptoms via the South Oaks Gambling Screen. Hierarchical multivariate regression analysis indicated that one domain of gambling cognitive distortions was associated significantly with gambling symptoms among the 342 symptomatic participants: gambling expectancy (β = 0.19, p < .05). The multivariate model explained 47% of the variance in the gambling symptoms. Conclusion: This study successfully contributed to the sparse research on university student gambling in Japan. Specifically, our results indicated a statistically significant relationship between gambling cognitive distortions and gambling disorder symptoms. These results can inform the development of preventive education and treatment for university students with gambling disorder in Japan. The report also describes needs for future research of university students with gambling disorder