Generation of retinal pigment epithelium from human induced pluripotent stem cells showed polarized secretion of VEGF and PEDF.

Age-related macular degeneration (AMD) is one of the major diseases that cause severe visual impairment in developed countries and is related to dysfunction of the retinal pigment epithelium (RPE). Human induced pluripotent stem cells (hiPSCs) have been drawing attention as a valuable source of RPE for basic research on or treatment of AMD. Here we investigated whether RPE could be generated from hiPSCs in Kawasaki Medical School. We maintained hiPSCs retaining pluripotent markers after a series of regular culture steps involving passaging, freezing, and thawing. hiPSCs were then cultured in RPE differentiation medium, and pigmented colonies were manually isolated for further differentiation into RPE. Differentiated cells formed pigmented cells with a typical RPE cobblestone appearance, abundant apical microvilli, adherens junctions, and tight junctions. Furthermore, generated pigmented cells expressed typical RPE marker genes, exhibited a barrier function, and secreted growth factors in a polarity-dependent manner similar to native RPE. These results indicate that RPE derived from hiPSCs in our facility can be used for in vitro and in vivo research

New Surgical Procedure for Pancreas Head

In this study, we demonstrate two new methods for pancreaticoduodenectomy (PD). One method is the mini‐laparotomic PD by Shuriken‐shaped umbilicoplasty with the real‐time moving window‘s method. The other method is the new pancreaticojejunostomy (PJ) by punctured stent slide guiding method (PSSGM). This procedure could be performed by complete mini‐laparotomy under direct vision, and the final major wound is only 2 cm of round navel. PSSGM prevents the difference of caliber between pancreatic anastomosis and the inside out of jejunal mucosa in theory. Ten cases of mini‐lap PD were successfully performed under new PJ anastomosis. The pancreatic leakage (PL) was only one case of ISGPF grade A, and its frequency was 9% (1/11). Our mini‐lap PD by Shuriken‐shaped umbilicoplasty might be a useful way for overcoming the obstacles about safety, complication risk, cosmetic demand, and medical cost compared to laparoscopic PD. Also, our new device of PJ reconstruction by PSSGM might be an easy and useful device for the prevention of PL

Tissue factor expression in human pterygium

Purpose: A pterygium shows tumor-like characteristics, such as proliferation, invasion, and epithelial–mesenchymal transition (EMT). Previous reports suggest that tissue factor (TF) expression is closely related to the EMT of tumor cells, and subsequent tumor development. In this study, we analyzed the expression and immunolocalization of TF in pterygial and normal conjunctival tissues of humans. Methods: Eight pterygia and three normal bulbar conjunctivas, surgically removed, were used in this study. Formalinfixed, paraffin-embedded tissues were submitted for immunohistochemical analysis with anti-TF antibody. Double staining immunohistochemistry was performed to assess TF and alpha-smooth muscle actin (α-SMA) or epidermal growth factor receptor (EGFR) expression in the pterygia. Results: Immunoreactivity for TF was detected in all pterygial tissues examined. TF immunoreactivity was localized in the cytoplasm of basal, suprabasal, and superficial epithelial cells. The number of TF-immunopositive cells in pterygial epithelial cells was significantly higher than in normal conjunctival epithelial cells (p<0.001). TF immunoreactivity was detected in α-SMA-positive or -negative pterygial epithelial cells. EGFR immunoreactivity was detected in pterygial epithelium, which was colocalized with TF. Conclusions: These results suggest that TF plays a potential role in the pathogenesis and development of a pterygium, and that TF expression might be involved through EMT-dependent and -independent pathways

Characterization of spliced leader trans-splicing in a photosynthetic rhizarian amoeba, Paulinella micropora, and its possible role in functional gene transfer

Paulinella micropora is a rhizarian thecate amoeba, belonging to a photosynthetic Paulinella species group that has a unique organelle termed chromatophore, whose cyanobacterial origin is distinct from that of plant and algal chloroplasts. Because acquisition of the chromatophore was quite a recent event compared with that of the chloroplast ancestor, the Paulinella species are thought to be model organisms for studying the early process of primary endosymbiosis. To obtain insight into how endosymbiotically transferred genes acquire expression competence in the host nucleus, here we analyzed the 5′ end sequences of the mRNAs of P. micropora MYN1 strain with the aid of a cap-trapper cDNA library. As a result, we found that mRNAs of 27 genes, including endosymbiotically transferred genes, possessed the common 5′ end sequence of 28–33 bases that were posttranscriptionally added by spliced leader (SL) trans-splicing. We also found two subtypes of SL RNA genes encoded by the P. micropora MYN1 genome. Differing from the other SL trans-splicing organisms that usually possess poly(A)-less SL RNAs, this amoeba has polyadenylated SL RNAs. In this study, we characterize the SL trans-splicing of this unique organism and discuss the putative merits of SL trans-splicing in functional gene transfer and genome evolution

The Effects of Exercise Training on Obesity-Induced Dysregulated Expression of Adipokines in White Adipose Tissue

Obesity is recognized as a risk factor for lifestyle-related diseases such as type 2 diabetes and cardiovascular disease. White adipose tissue (WAT) is not only a static storage site for energy; it is also a dynamic tissue that is actively involved in metabolic reactions and produces humoral factors, such as leptin and adiponectin, which are collectively referred to as adipokines. Additionally, because there is much evidence that obesity-induced inflammatory changes in WAT, which is caused by dysregulated expression of inflammation-related adipokines involving tumor necrosis factor-α and monocyte chemoattractant protein 1, contribute to the development of insulin resistance, WAT has attracted special attention as an organ that causes diabetes and other lifestyle-related diseases. Exercise training (TR) not only leads to a decrease in WAT mass but also attenuates obesity-induced dysregulated expression of the inflammation-related adipokines in WAT. Therefore, TR is widely used as a tool for preventing and improving lifestyle-related diseases. This review outlines the impact of TR on the expression and secretory response of adipokines in WAT

iPS 細胞由来網膜色素上皮細胞移植のためのNaIO3誘因網膜色素上皮変性げっ歯類モデルの特性評価

網膜色素上皮（RPE）は視細胞維持に重要な役割を担い，その機能低下は加齢黄斑変性等の原因となる．近年，健常なRPE を補充するRPE 移植が注目されており，臨床応用も開始された．前臨床研究における動物実験にあたり，過去にもげっ歯類のヨウ素酸ナトリウム（NaIO3）誘因RPE 変性モデルの研究はあるが，げっ歯類は黄斑部を欠くため最適でない．黄斑を有する非ヒト霊長類における薬剤全身投与は両眼の失明を招くため動物愛護の観点から不適切であり，現在適切なモデルが存在しない．従って，我々はサルを用いた移植実験を念頭に置きNaIO3の硝子体内投与によりラット片眼にRPE 変性を誘発可能か研究した．また，NaIO3誘因RPE 変性モデルにヒト多能性幹細胞由来RPE（hiPS-RPE）を網膜下移植し視細胞の保護効果を検討した．8-10週齢のオスのWistar ラットの片眼に0.005，0.01，0.02 mg/μl のNaIO3を硝子体内投与した．経時的にHE 染色にて網膜外層厚と網膜内層厚を計測し，透過型電子顕微鏡で形態学的評価を行った．NaIO3投与後2日目にhiPS-RPE 懸濁液を網膜下に移植し，組織学的評価を行った．0.005 mg/μl 投与群では網膜内層・外層厚ともに変化はなく，0.02 mg/μl 投与群では網膜全層が障害され，0.01 mg/μl 投与群では網膜外層のみが障害された．RPE の形態学的変化はNaIO3投与後3日目から出現し，以降網膜外層厚が急速に菲薄化した．いずれの投与群も非投与眼に影響はなかった．hiPS-RPE 移植眼では，網膜外層厚が厚い傾向にあった．まとめると，NaIO3片眼硝子体内投与により，げっ歯類片眼RPE 障害モデル作製に成功した．hiPS-RPE 網膜下移植により視細胞保護効果を得られた．本方法はRPE 移植の前臨床研究の動物モデルの基礎として有用である．The retinal pigment epithelium (RPE) plays an important role in maintaining photoreceptors, and RPE dysfunction causes eye diseases such as age-related macular degeneration (AMD). RPE transplantation has garnered much attention in recent years as a way to replenish healthy RPE, and clinical applications for RPE transplantation in patients with AMD have been reported. Preclinical studies use animal models to examine treatment efficacy, however, whether conventional methods with animal models are suitable for examining RPE transplantation is controversial. Systemic administration of sodium iodate (NaIO3) has beenshown to induce RPE degeneration in rodent models, recapitulating morphological atrophy in corresponding areas. However, as rodents lack a macular region, these studies cannot precisely evaluate the effects of systemic administration. The induction of blindness in both eyes in nonhuman primates bearing a macular region is also inappropriate in terms of animal ethics. With this in mind, we investigated whether intravitreal injection of NaIO3 can selectively induce a single-eye impairment in an RPE animal model. Additionally, we investigated whether subretinal transplantation of human induced pluripotent stem cells-derived RPE (hiPS-RPE) can protect against NaIO3-induced RPE degeneration.The intravitreal injection of NaIO3 was performed in 8-10 week-old male Wistar rats with rats being grouped by dose (0.005, 0.01, and 0.02 mg/μl). We measured outer nuclear layer (ONL) thickness and inner nuclear layer (INL) thickness at 8 retinal points by hematoxylin-eosin staining and evaluated RPE morphology by transmission electron microscopy sequentially (3, 7, 14, and 28 days after injection). In addition, hiPSC-RPE cell suspension was injected into the subretinal space 2 days after NaIO3 administration and histological evaluation was performed sequentially (7, 14, and 28 days after NaIO3 injection).The results showed no significant changes in INL and ONL thicknesses in the 0.005 mg/μl group. Retinal thicknesses in the 0.02 mg/μl group had almost completely degenerated. In the 0.01 mg/μl group, ONL thickness had almost completely degenerated, but, in contrast INL thickness was preserved as well as 0.005 mg/μl injection group. RPE morphological changes and TUNEL-positive cells appeared at 3 days after NaIO3 injection, followed by rapid thinning of ONL thickness. None of the retinal thickness of contralateral eye in all groups showed any changes. Additionally, ONL thicknesses in the transplanted eyes were significantly greater than those in the nontransplanted and sham-surgery groups

Prediction of functional recovery after revascularization using quantitative gated myocardial perfusion SPECT: a multi-center cohort study in Japan

金沢大学医薬保健研究域医学系Backgrounds: Prediction of left ventricular functional recovery is important after myocardial infarction. The impact of quantitative perfusion and motion analyses with gated single-photon emission computed tomography (SPECT) on predictive ability has not been clearly defined in multi-center studies. Methods: A total of 252 patients with recent myocardial infarction (n = 74) and old myocardial infarction (n = 175) were registered from 25 institutions. All patients underwent resting gated SPECT using 99mTc-hexakis-2-methoxy-isobutyl isonitrile (MIBI) and repeated the study after revascularization after an average follow-up period of 132 ± 81 days. Visual and quantitative assessment of perfusion and wall motion were performed in 5,040 segments. Results: Non-gated segmental percent uptake and end-systolic (ES) percent uptake were good predictors of wall motion recovery and significantly differed between improved and non-improved groups (66 ± 17% and 55 ± 18%, p < 0.0001 for non-gated; 64 ± 16% and 51 ± 17% for ES percent uptake, p < 0.0001). The area under the curve of receiver operating characteristics curve for non-gated percent uptake, ES percent uptake, end-diastolic percent uptake and visual perfusion defect score was 0.70, 0.71, 0.61, and 0.56, respectively. Sensitivity and specificity of percent uptake were 68% and 64% for non-gated map and 80% and 52% for ES percent uptake map. An optimal threshold for predicting segmental improvement was 63% for non-gated and 52% for ES percent uptake values. Conclusion: Segmental 99mTc-MIBI uptake provided a useful predictor of wall motion improvement. Application of quantitative approach with non-gated and ES percent uptake enhanced predictive accuracy over visual analysis particularly in a multi-center study. © 2008 Springer-Verlag

Creation and characterization of Japanese standards for myocardial perfusion SPECT: database from the Japanese Society of Nuclear Medicine Working Group

金沢大学大学院医学系研究科がん制御

Current Performance and On-Going Improvements of the 8.2 m Subaru Telescope

An overview of the current status of the 8.2 m Subaru Telescope constructed and operated at Mauna Kea, Hawaii, by the National Astronomical Observatory of Japan is presented. The basic design concept and the verified performance of the telescope system are described. Also given are the status of the instrument package offered to the astronomical community, the status of operation, and some of the future plans. The status of the telescope reported in a number of SPIE papers as of the summer of 2002 are incorporated with some updates included as of 2004 February. However, readers are encouraged to check the most updated status of the telescope through the home page, http://subarutelescope.org/index.html, and/or the direct contact with the observatory staff.Comment: 18 pages (17 pages in published version), 29 figures (GIF format), This is the version before the galley proo

ケイブ コユウ カンカク トレーニング ガ ジュウシン ドウヨウ ニ オヨボス エイキョウ ニ ツイテ ノ ケンキュウ

PURPOSE: The purpose of this study was to investigate the effectiveness of cervicocephalic kinesthetic training on postural stability and neck joint position sense in healthy young adults. METHOD: 33 subjects were recruited for this study and randomly divided into training （n＝16） and control groups （n＝17）. The training group performed cervicocephalic kinesthetic training for 5 minutes per day for 10 days and the control group performed the same training but with eyes closed to eliminate visual feedback. All training sessions for both groups were performed under supervision. Pre and post cervical joint position sense and sensory organization testing （SOT） were measured on both groups. RESULT: Significant differences were found in cervical joint position sense and SOT condition 6 in the training group. No significant differences were found in SOT condition1-5 in both groups and cervical joint position in the control group. CONCLUSION: The improvements of postural stability and cervical joint position sense after 10 days of cervicocephalic kinesthetic training suggest that cervical joint position sense may may be beneficial for postural stability in healthy young adults