Effects of TGF-β1 and IGF-1 on proliferation of human nucleus pulposus cells in medium with different serum concentrations

BACKGROUND: The low proliferative viability of human nucleus pulposus(NP) cells is considered as a cause of intervertebral discs degeneration. Growth factors, such as TGF-β1 and IGF-1, have been implicated in cell proliferation and matrix synthesis. OBJECTIVE: To investigate the dose-response and time-course effect of transforming growth factorβ1(TGF-β1) and insulin-like growth factor-1(IGF-1) on proliferation of NP cells. STUDY DESIGN: 3-(4,5-dimethylthiazolyl)-2,5-diphenyl-tetrazolium bromide (MTT) is reduced by dehydrogenase in mitochondria of live cells. The proliferative viability of cells corresponds to the amount of MTT reduced, which is measured with an enzyme-linked immunosorbent assay plate reader. In this study, we assessed dose- and time-dependent effects of NP cells to TGF-β1 and IGF-1 in medium with different serum concentrations by MTT assay. METHODS: After release of informed consent, tissue samples of NP were obtained from anterior surgical procedures performed on five donors with idiopathic scoliosis. Isolated cells were cultured in F12 medium supplemented with 10% fetal bovine serum(FBS). Cells were seeded in 96-well plates at 1 × 10(3 )cells/well. After synchronization, medium was replaced by F12 containing 1% or 10% FBS with either single or combination of TGF-β1 and IGF-1. Dose-response and time-course effect were examined by MTT assay. RESULTS: In the presence of 1% FBS, the response to IGF-1 was less striking, whereas TGF-β1 had a remarkably stimulating effect on cell proliferation. In 10% FBS, both of the two growth factors had statistical significant mitogenic effects, especially TGF-β1. The dose-dependent effect of TGF and IGF on cell proliferation was found within different concentrations of each growth factor(TGF-β1 1–10 μg/L, IGF-1 10–100 μg/L). The time-course effect showed a significant elevation three days later. CONCLUSION: TGF-β1 and IGF-1 were efficient to stimulate cell proliferation of human NP cells in vitro with a dose- and time-dependent manner. These results support the therapeutic potentials of the two growth factors in the treatment of disc degeneration

Nitrosative and Oxidative Stresses Contribute to Post-Ischemic Liver Injury Following Severe Hemorrhagic Shock: The Role of Hypoxemic Resuscitation

Purpose: Hemorrhagic shock and resuscitation is frequently associated with liver ischemia-reperfusion injury. The aim of the study was to investigate whether hypoxemic resuscitation attenuates liver injury. Methods: Anesthetized, mechanically ventilated New Zealand white rabbits were exsanguinated to a mean arterial pressure of 30 mmHg for 60 minutes. Resuscitation under normoxemia (Normox-Res group, n = 16, PaO2 = 95–105 mmHg) or hypoxemia (Hypox-Res group, n = 15, PaO 2 = 35–40 mmHg) followed, modifying the FiO 2. Animals not subjected to shock constituted the sham group (n = 11, PaO 2 = 95–105 mmHg). Indices of the inflammatory, oxidative and nitrosative response were measured and histopathological and immunohistochemical studies of the liver were performed. Results: Normox-Res group animals exhibited increased serum alanine aminotransferase, tumor necrosis factor- alpha, interleukin (IL)-1b and IL-6 levels compared with Hypox-Res and sham groups. Reactive oxygen species generation, malondialdehyde formation and myeloperoxidase activity were all elevated in Normox-Res rabbits compared with Hypox-Res and sham groups. Similarly, endothelial NO synthase and inducible NO synthase mRNA expression was up-regulated and nitrotyrosine immunostaining increased in animals resuscitated normoxemically, indicating a more intense nitrosative stress. Hypox-Res animals demonstrated a less prominent histopathologic injury which was similar to sham animals. Conclusions: Hypoxemic resuscitation prevents liver reperfusion injury through attenuation of the inflammatory respons

柿樹の栄養生理的研究(第 1 報) : 無機成分の体内分布ならびに吸収量の季節的変化について(農学部門)

1.柿栽培における施肥の合理化を進める資料をえる目的で, 1957∿58年にわたり富有, 次郎, 会津身不知の成木を供試して, 無機成分の体内分布, 各部位の含有率および吸収量の季節的消長を調査した。2.枝葉, 果実の新成組織内の含有率は旧生部より高く, 生育時期別にみた場合に, 発育初期の含有率は後期に比べて高い傾向が認められた。3.葉内含有率はNはほぼ生育初期に高く生育期間中は多少の増減はみられるが大きな変化はなく着果の影響も少ない。Pは生育当初に高く以後漸減の傾向を示すが, 幼果の生育が急速に進む頃に着果枝内含有率が低下し6月下旬頃にその傾向が著しい。KについてはN, Pに比べて当初高くなく季節的消長はNより大きいものであつたが, 着果によつて会津身不知では減少するが, 富有では必ずしも減少せずむしろ8月中旬頃ではかえつて高い結果を認めた。4.新梢内では生育当初の含有率は高く以後漸減しているが, Nについて着果の影響は少なく, P, Kについては着果によつて多少低下する傾向が認められた。5.果実内ではNは花蕾発生初期より果実発育第I期頃まで含有率が高く以後漸減する傾向があり, Pは初期に高く第I期頃よりわずかに減少し, Kは当初Nより少なく第I期終期頃より増加しN含有率より高くなるが, 生長後期にはやや減少の傾向が認められた。6.葉内吸収量については, N, Kでは吸収量も多くその増減も大きいが, Pは吸収量およびその変化も少なかつた。その季節的消長は各要素とも生長にともなつて吸収量は増加するが, 6月中下旬に一時やや低下しその後ふたたび増大するが, 8月下旬以降は漸減する傾向を示した。このことは展葉後の新梢の伸長および果実肥大に由来するものと思われるが, 葉内吸収量は両品種とも着果枝は生育当初ほぼ無着果枝より高いが, 生長後期では富有では着果枝が会津身不知では無着果枝が高い傾向が認められた。7.果実内吸収量は果実肥大とともに増大し, 8月中下旬にやや劣り9月中旬以降ふたたび増加するが, 特にKはN, Pに比べてこの傾向が著しい。8.枝葉, 果実を含む新梢1本当たりのN, P, Kの吸収量は, 展葉より7月中旬頃まで著しく増加しやがて8月下旬までやや減少するが, 9月初旬頃よりふたたび増大する。新梢生長期および生育後期の増加が大きく, K, Pは9月下旬頃に最高となり, Nは6月下旬∿7月中旬に高くなつた。会津身不知では吸収量の最大となる時期が富有より約1カ月程度おくれる傾向がある。9.生長周期を新梢伸長期, 果実発育第I期, 第II期, 第III期に区分して, 各時期別の1日当たり平均吸収量をみると, 両品種とも新梢伸長期および果実発育第I期に吸収量が大となり, 第II, III期ではやや低く一時とどまる傾向が認められた。その吸収量最大期は富有では着果枝が第I期, 無着果枝が新梢伸長期となり, 会津身不知では着果枝が新梢伸長期, 無着果枝では第I期であつた。In the present study the authers investigated he internal distribution and seasonal absor ption of N, P and K in the Japanese persimmons (Fuyu, Jiro and Aizumishirazu), using mature trees. The leaves and fruits as the new tissues were highest in the content of any constituent among the another organs and the contents of all constitution decreased in the latter period of growing season. Concerning a seasonal change of the amounts of N, P and K in the fruit were increased with development of fruit towards the ripening period, however at their second growing period the rate of absorption tended to be slow down temporary. In the shoot including leaves, twig and fruits, the amounts of absorption were abundant in the bearing shoot than in the non-bearing, and they increased highly with the development of leaves and fruits in the middle of July, but the maximum absorption of P and K was observed in the end of september, and N absorption was in the middle of July. The ratio of N, P and K absorbed by fruit was 10 : 4.6 : 13.6 by Fuyu and 10 : 5.9 : 14.2 by Aizumishirazu in the middle of september, and 10 : 6.2 : 18.4 by Fuyu in the end of October. In the middle of september, the amouuts of N, P and K absorbed by the current shoot including leaves, twing and fruits was 243mg : 76mg : 288mg by Fuyu and 179mg : 54mg : 204mg by Aizumishirazu at the bearing shoot, but in the non-bearing shoot 184mg : 31mg : 124mg by Fuyu and 132mg : 22mg : 130mg by Aizumishirazu

Human DEF6 deficiency underlies an immunodeficiency syndrome with systemic autoimmunity and aberrant CTLA-4 homeostasis

Immune responses need to be controlled tightly to prevent autoimmune diseases, yet underlying molecular mechanisms remain partially understood. Here, we identify biallelic mutations in three patients from two unrelated families in differentially expressed in FDCP6 homolog (DEF6) as the molecular cause of an inborn error of immunity with systemic autoimmunity. Patient T cells exhibit impaired regulation of CTLA-4 surface trafficking associated with reduced functional CTLA-4 availability, which is replicated in DEF6-knockout Jurkat cells. Mechanistically, we identify the small GTPase RAB11 as an interactor of the guanine nucleotide exchange factor DEF6, and find disrupted binding of mutant DEF6 to RAB11 as well as reduced RAB11+CTLA-4+ vesicles in DEF6-mutated cells. One of the patients has been treated with CTLA-4-Ig and achieved sustained remission. Collectively, we uncover DEF6 as player in immune homeostasis ensuring availability of the checkpoint protein CTLA-4 at T-cell surface, identifying a potential target for autoimmune and/or cancer therapy.Mechanistically, we identify the small GTPase RAB11 as an interactor of the guanine nucleotide exchange factor DEF6, and find disrupted binding of mutant DEF6 to RAB11 as well as reduced RAB11+CTLA-4+ vesicles in DEF6-mutated cells. One of the patients has been treated with CTLA-4-Ig and achieved sustained remission. Collectively, we uncover DEF6 as player in immune homeostasis ensuring availability of the checkpoint protein CTLA-4 at T-cell surface, identifying a potential target for autoimmune and/or cancer therapy

遮光が桃樹の生育, 光合成及び葉の内部構造に及ぼす影響(農学部門)

5年生高陽白桃を供試して地上部を葭簀で遮光処理を行い, 枝葉の伸長量, 同化量, 葉の内部構造等を生長に伴つて季節的に比較調査し, 生長末期に円陣法によつて根群の分布を観察し, 併せて地上部, 地下部の生体重を比較測定した。新梢伸長は遮光によつて抑制され, 遮光程度が進むと枯死した。葉の生長では横径よりも縦径に及ぼす影響が大きく, 遮光程度が少ない場合には徒長して葉面積が大きく, 葉肉組織の厚さは薄くなり柵状, 海綿状組織は粗であつた。遮光によつて同化作用は抑制され, 無処理区に対して2/5∿1/10程度に低下し, 曇雨天日には晴天日に比して減少程度が顕著であつた。遮光の処理は根の生長にも悪影響を及ぼし, 伸長分布は水平的にも垂直的にも抑制され, 特に細根量が著しく減少した。1. Experiments were carried out in 1959 in order to study the effects of covering 5 years old peach trees with bamboo blind through the growing season, from the view point of the shoot elongation, photosynthesis, and the internal structure of leaves and roots system. 2. As the results, the shoot elongation and leaf thickness were decreased by shading, and the internal structure of leaves are non compacted to compared with the natural leaves. The natural range of roots were observed in radius of 3m and at a depth of 120cm, while the roots of covering plot were not observed more than radius of 2m and at a depth of 90cm, especially the fibrous roots were decreased. The photosynthesis were decreasad by shading, and the influence of covering became powerful in cloudy more than fine days

Design and baseline characteristics of the finerenone in reducing cardiovascular mortality and morbidity in diabetic kidney disease trial

Background: Among people with diabetes, those with kidney disease have exceptionally high rates of cardiovascular (CV) morbidity and mortality and progression of their underlying kidney disease. Finerenone is a novel, nonsteroidal, selective mineralocorticoid receptor antagonist that has shown to reduce albuminuria in type 2 diabetes (T2D) patients with chronic kidney disease (CKD) while revealing only a low risk of hyperkalemia. However, the effect of finerenone on CV and renal outcomes has not yet been investigated in long-term trials. Patients and Methods: The Finerenone in Reducing CV Mortality and Morbidity in Diabetic Kidney Disease (FIGARO-DKD) trial aims to assess the efficacy and safety of finerenone compared to placebo at reducing clinically important CV and renal outcomes in T2D patients with CKD. FIGARO-DKD is a randomized, double-blind, placebo-controlled, parallel-group, event-driven trial running in 47 countries with an expected duration of approximately 6 years. FIGARO-DKD randomized 7,437 patients with an estimated glomerular filtration rate >= 25 mL/min/1.73 m(2) and albuminuria (urinary albumin-to-creatinine ratio >= 30 to <= 5,000 mg/g). The study has at least 90% power to detect a 20% reduction in the risk of the primary outcome (overall two-sided significance level alpha = 0.05), the composite of time to first occurrence of CV death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for heart failure. Conclusions: FIGARO-DKD will determine whether an optimally treated cohort of T2D patients with CKD at high risk of CV and renal events will experience cardiorenal benefits with the addition of finerenone to their treatment regimen. Trial Registration: EudraCT number: 2015-000950-39; ClinicalTrials.gov identifier: NCT02545049