Generating Vegfr3 reporter transgenic mouse expressing membrane-tagged Venus for visualization of VEGFR3 expression in vascular and lymphatic endothelial cells

Vascular endothelial growth factor receptor 3 (Vegfr3) has been widely used as a marker for lymphatic and vascular endothelial cells during mouse embryonic development and in adult mouse, making it valuable for studying angiogenesis and lymphangiogenesis under normal and pathological conditions. Here, we report the generation of a novel transgenic (Tg) mouse that expresses a membrane-localized fluorescent reporter protein, Gap43-Venus, under the control of the Vegfr3 regulatory sequence. Vegfr3-Gap43-Venus BAC Tg recapitulated endogenous Vegfr3 expression in vascular and lymphatic endothelial cells during embryonic development and tumor development. Thus, this Tg mouse line contributes a valuable model to study angiogenesis and lymphangiogenesis in physiological and pathological contexts

カンイガタ ジキシキ ショクジ レキホウ シツモンヒョウ（BDHQ） オ モチイタ ガイライ トウニョウビョウ エイヨウ シドウ タイショウシャ チュウシュツホウ ニカンスル ケントウ

【背景】地域に開かれた無床診療所には、管理栄養士はほとんど常勤しておらず、栄養指導が必要な生活習慣 病患者に対する指導でさえも十分に実施できていない現状がある。また、糖尿病栄養指導対象者抽出については、医 師の主観（治療歴、患者背景、患者の性格等）や血液検査値等の客観的指標のみを参考にしており、必ずしも食事調 査結果に基づいた抽出をしている訳ではない。そのため、実際に食習慣が乱れており、糖尿病栄養指導の必要性が高 い患者に対して効率良く栄養指導ができているとは言い難い。 【目的】無床診療所における外来糖尿病患者の治療効果を向上させるため、BDHQ から得られた食事調査結果を活 用した新しい外来糖尿病栄養指導対象者抽出法を確立すること。 【方法】M 医院（大阪府阪南市）に通院している2 型糖尿病患者のうち、研究の趣旨に同意が得られた109 名（男 性77 名、女性32 名、平均年齢64.7±7.4 歳）にBDHQ を実施し、ある1 日の平均的な「栄養素摂取量」を算出し た。次いで当該患者カルテの検査値（HbA1c、クレアチニン等）を記録し、食事調査結果と組み合わせた栄養指導 対象者抽出基準を検討した。 【結果および考察】糖尿病食事療法のうち、栄養基準が大きく変化する糖尿病性腎症第3 期B に該当する患者は、 109 名中25 名であった。このうち、BDHQ から得られたたんぱく質摂取量が1.0g/kg 標準体重/日を上まっている 患者は17 名であり、特にこれらの患者は管理栄養士による詳細な栄養指導を受ける必要性が高いことが考えられる。 BDHQ は集団での妥当性はある程度証明されているが、個人での妥当性は証明されていない。そのため得られた結 果の解釈には慎重な配慮を要する。しかし、少なくとも今回のように、無床診療所において管理栄養士による詳細な 栄養指導を受ける必要性があるか否かを大雑把に判定するような目的には最適であろうと考えられた。Purpose: To improve therapeutic efficacy of ambulatory patients with diabetes in non-bed clinics and to establish a new method of identifying subjects at risk of diabetes related complications using a dietary survey obtained from a brief self administered diet history questionnaire（BDHQ）. Methods: Among patients with type 2 diabetes attending M clinic（Osaka, Japan）, 109 gave informed consent（male 77, female 32, mean age 64.7 ± 7.4 years）. A BDHQ was administered to calculate the average “nutrient intake” per day. The test results in the patients’ medical records（HbA1c, creatinine, etc.） were recorded. The criteria for identifying patients requiring nutritional guidance were also examined in conjunction with the dietary surveys. Results: For 25 of the participants, their nutritional standards put them at risk of third phase B diabetic nephropathy. Of these, the protein intake of 17, as calculated from the BDHQ, was >1.0 g/kg/day. It was considered essential that these patients receive detailed nutritional guidance from registered dietitians. Although the relevance of administering group BDHQ is to some extent proved, the benefit for the individual has not been demonstrated. The interpretation of the results must, therefore, be viewed with caution. However, there are indicators that suggest detailed nutritional guidance by registered dietitians at non bed clinics would be advisable. Keywords: Non bed Clinic, I

Fluorescence and Bioluminescence Imaging of Angiogenesis in Flk1-Nano-lantern Transgenic Mice

Angiogenesis is important for normal development as well as for tumour growth. However, the molecular and cellular mechanisms underlying angiogenesis are not fully understood, partly because of the lack of a good animal model for imaging. Here, we report the generation of a novel transgenic (Tg) mouse that expresses a bioluminescent reporter protein, Nano-lantern, under the control of Fetal liver kinase 1 (Flk1). Flk1-Nano-lantern BAC Tg mice recapitulated endogenous Flk1 expression in endothelial cells and lymphatic endothelial cells during development and tumour growth. Importantly, bioluminescence imaging of endothelial cells from the aortic rings of Flk1-Nano-lantern BAC Tg mice enabled us to observe endothelial sprouting for 18 hr without any detectable phototoxicity. Furthermore, Flk1-Nano-lantern BAC Tg mice achieved time-lapse luminescence imaging of tumour angiogenesis in freely moving mice with implanted tumours. Thus, this transgenic mouse line contributes a unique model to study angiogenesis within both physiological and pathological contexts

Critical Roles of the WASP N-Terminal Domain and Btk in LPS-Induced Inflammatory Response in Macrophages

While Wiskott-Aldrich syndrome protein (WASP) plays critical roles in TCR signaling as an adaptor molecule, how it transduces innate immune signals remains to be elucidated. To investigate the roles of WASP in innate immune cells, we established bone marrow-derived macrophage (BMDM) cell lines from WASP15 transgenic (Tg) mice overexpressing the WASP N-terminal region (exons 1–5). Upon LPS stimulation, WASP15 Tg BMDM cell lines produce lower levels of inflammatory cytokines, such as TNF-α, IL-6, and IL-12p40 than the wild-type BMDM cell line. In addition, the production of nitric oxide by WASP15 Tg BMDM cells in response to LPS and IFN-γ was significantly impaired. Furthermore, we uncovered that the WASP N-terminal domain associates with the Src homology (SH) 3 domain of Bruton's tyrosine kinase (Btk). Overexpression of the WASP N-terminal domain diminishes the extent of tyrosine phosphorylation of endogenous WASP in WASP15 Tg BMDM cells, possibly by interfering with the specific binding between endogenous WASP and Btk during LPS signaling. These observations strongly suggest that the interaction between WASP N-terminal domain and Btk plays important roles in the LPS signaling cascade in innate immunity

イギリスにおける政策の導入と変遷：1988 年から2013 年の25 年間にわたる教師たちへの影響

As policies for national education become increasingly contested, professional development of teachers needs to include critical understanding of the political context within which they work. That was the rationale for my recent book: Politics and the Primary Teacher (Routledge 2012). This seminar is based on research undertaken in the UK on the role of primary teachers, but will also identify distinct developments in secondary schooling. Consideration will be given to curriculum, pedagogy, teacher status and local and central governance and administration of schools in both the primary and secondary sectors, including quality assurance

Changes in the chemical compositions of leaf litter in the canopy of a Japanese cedar plantation

Canopy leaf litter is typically unevenly distributed due to interception by branches and epiphytes and/or to attachment to branches and tree trunks. Japanese cedar (Cryptomeria japonica) trees hold a large amount of canopy litter that stays attached to foliage and branches for several years before litterfall. Litter retention in the canopy is predicted to affect the chemical composition of litterfall and throughfall. We analyzed the chemical compositions of water-soluble materials washed from the leaves and those of the leaf residues with different leaf litter ages to clarify the effect of leaf-litter retention in the canopy of Japanese cedar trees. Chemical components of the leaf litter after the washing procedure show temporal changes in total carbon, total nitrogen, and carbon/nitrogen ratio just after senescence and prior to litterfall. Higher inorganic ion concentrations in soluble materials from old dead leaves (ODL) dead for more than 2 years suggest the importance of ODL in leaching and catching chemical components in the canopy. Our results show that Japanese cedar canopies retain the various qualities of leaf litter, which are produced by senescence, leaching, and catching dry deposition, and that a large amount of dissolved components leach from dead leaves that are still attached to the canopy

Determination of a pain substance produced by the photodegradation of dacarbazine

The anticancer drug, dacarbazine, is known to be photosensitive, and its photodegradation products have been pointed out as the causes of side effects including local venous pain of injection site. In this study, we attempted to clarify the causative substance of pain after photodegradation of dacarbazine. We synthesized five photodegradation products of dacarbazine; dimethylamine, 5-diazoimidazole-4-carboxamide (Diazo-IC), 4-carbamoylimidazolium-5-olate, 4-carbamoyl-2-(4-carbamoylimidazol-5-ylazo) imidazolium-5-olate and 2-azahypoxanthine, and examined pain reactions induced by these substances in mice. Mice were intraperitoneally administered each photodegradation product, then number of stretching reactions or writhing reactions as types of pain behaviors was counted. Only Diazo-IC clearly induced the pain reactions in mice in a concentration-dependent manner: the other products caused no pain reaction. The pain threshold of Diazo-IC in mice was estimated at between 0.1 mg/ml and 0.2 mg/ml. While diclofenac sodium significantly reduced acetic acid-induced pain reactions in mice, it did not influence the reactions induced by Diazo-IC. This result suggests that Diazo-IC-induced pain reactions represent a different mechanism from acetic acid-induced inflammatory pain. Degradation rate constant of 0.1 mg/ml of dacarbazine solution was 10 times larger than 1 mg/ml of dacarbazine. Dacarbazine solution for drip infusion should be sufficiently shielded from light

Clinical features of patients with acute coronary syndrome during the COVID-19 pandemic

International audienceAlthough a reduction in hospital admissions of acute coronary syndromes (ACS) patients has been observed globally during the coronavirus disease 2019 (COVID-19) pandemic, clinical features of those patients have not been fully investigated. The aim of the present analysis is to investigate the incidence, clinical presentation, and outcomes of patients with ACS during the COVID-19 pandemic. We performed a retrospective analysis of consecutive patients who were admitted for ACS at our institution between March 1 and April 20, 2020 and compared with the equivalent period in 2019. Admissions for acute myocardial infarction (AMI) reduced by 39.5% in 2020 compared with the equivalent period in 2019. Owing to the emergency medical services (EMS) of our region, all time components of ST-elevated myocardial infarction care were similar during the COVID-19 outbreak as compared with the previous year's dataset. Among the 106 ACS patients in 2020, 7 patients tested positive for COVID-19. Higher incidence of type 2 myocardial infarction (29% vs. 4%, p = 0.0497) and elevated D-dimer levels (5650 μg/l [interquartile range (IQR) 1905-13,625 μg/l] vs. 400 μg/l [IQR 270-1050 μg/l], p = 0.02) were observed in COVID-19 patients. In sum, a significant reduction in admission for AMI was observed during the COVID-19 pandemic. COVID-19 patients were characterized by elevated D-dimer levels on admission, reflecting enhanced COVID-19 related thrombogenicity. The prehospital evaluation by EMS may have played an important role for the timely revascularization for STEMI patients