Comparative Genomics Identifies the Mouse BMP3 Promoter and an Upstream Evolutionary Conserved Region (ECR) in Mammals.

The Bone Morphogenetic Protein (BMP) pathway is a multi-member signaling cascade whose basic components are found in all animals. One member, BMP3, which arose more recently in evolution and is found only in deuterostomes, serves a unique role as an antagonist to both the canonical BMP and Activin pathways. However, the mechanisms that control BMP3 expression, and the cis-regulatory regions mediating this regulation, remain poorly defined. With this in mind, we sought to identify the Bmp3 promoter in mouse (M. musculus) through functional and comparative genomic analyses. We found that the minimal promoter required for expression in resides within 0.8 kb upstream of Bmp3 in a region that is highly conserved with rat (R. norvegicus). We also found that an upstream region abutting the minimal promoter acts as a repressor of the minimal promoter in HEK293T cells and osteoblasts. Strikingly, a portion of this region is conserved among all available eutherian mammal genomes (47/47), but not in any non-eutherian animal (0/136). We also identified multiple conserved transcription factor binding sites in the Bmp3 upstream ECR, suggesting that this region may preserve common cis-regulatory elements that govern Bmp3 expression across eutherian mammals. Since dysregulation of BMP signaling appears to play a role in human health and disease, our findings may have application in the development of novel therapeutics aimed at modulating BMP signaling in humans

On the Emerging Role of the Taste Receptor Type 1 (T1R) Family of Nutrient-Sensors in the Musculoskeletal System.

The special sense of taste guides and guards food intake and is essential for body maintenance. Salty and sour tastes are sensed via ion channels or gated ion channels while G protein-coupled receptors (GPCRs) of the taste receptor type 1 (T1R) family sense sweet and umami tastes and GPCRs of the taste receptor type 2 (T2R) family sense bitter tastes. T1R and T2R receptors share similar downstream signaling pathways that result in the stimulation of phospholipase-C-β2. The T1R family includes three members that form heterodimeric complexes to recognize either amino acids or sweet molecules such as glucose. Although these functions were originally described in gustatory tissue, T1R family members are expressed in numerous non-gustatory tissues and are now viewed as nutrient sensors that play important roles in monitoring global glucose and amino acid status. Here, we highlight emerging evidence detailing the function of T1R family members in the musculoskeletal system and review these findings in the context of the musculoskeletal diseases sarcopenia and osteoporosis, which are major public health problems among the elderly that affect locomotion, activities of daily living, and quality of life. These studies raise the possibility that T1R family member function may be modulated for therapeutic benefit

Adrenocorticotropic hormone at pathophysiological concentration modulates the proliferation and differentiation of bone cells

SummaryBackground/purposeAdrenocorticotropic hormone (ACTH) plays a vital role in maintaining the function of the hypothalamic–pituitary–adrenal axis. Recent studies have demonstrated that ACTH directly affects the proliferation and differentiation of bone cells. However, the ACTH concentrations used in these studies appear to be markedly higher than the physiological concentrations. Here, we investigated whether ACTH at pathophysiological concentration affects the proliferation and differentiation of osteoblasts and osteoclasts.Materials and methodsWe evaluated the effect of ACTH at pathophysiological concentration on osteoclasts using tartrate-resistant acid phosphatase staining and on osteoblasts using alkaline phosphatase activity assay. Additionally, we conducted reverse transcriptase-polymerase chain reaction analysis.ResultsWe found that at pathophysiological concentration, ACTH does not affect osteoblast proliferation and inhibits osteoblast differentiation. Moreover, we showed that at pathophysiological concentration, ACTH does not affect the proliferation of bone marrow macrophages, but promotes differentiation of osteoclasts and induces expression of genes involved in bone resorption.ConclusionTaken together, our findings suggest that ACTH modulates the proliferation and differentiation of bone cells in vitro at pathophysiological concentration

BMP3 Suppresses Osteoblast Differentiation of Bone Marrow Stromal Cells Via Interaction With Acvr2b.

Enhancing bone morphogenetic protein (BMP) signaling increases bone formation in a variety of settings that target bone repair. However, the role of BMP in the maintenance of adult bone mass is not well understood. Targeted disruption of BMP3 in mice results in increased trabecular bone formation, whereas transgenic overexpression of BMP3 in skeletal cells leads to spontaneous fracture, consistent with BMP3 having a negative role in bone mass regulation. Here we investigate the importance of BMP3 as a mediator of BMP signaling in the adult skeleton. We find that osteoblasts (OBL) and osteocytes are the source of BMP3 in adult bone. Using in vitro cultures of primary bone marrow stromal cells, we show that overexpression of BMP3 suppresses OBL differentiation, whereas loss of BMP3 increases colony-forming unit fibroblasts and colony-forming unit OBL. The ability of BMP3 to affect OBL differentiation is due to its interaction with activin receptor type 2b (Acvr2b) because knockdown of endogenous Acvr2b in bone marrow stromal cells reduces the suppressive effect of BMP3 on OBL differentiation. These findings best fit a model in which BMP3, produced by mature bone cells, acts to reduce BMP signaling through Acvr2b in skeletal progenitor cells, limiting their differentiation to mature OBL. Our data further support the idea that endogenous BMPs have a physiological role in regulating adult bone mass

Expression of Ascorbate Peroxidase Derived from Cyanidioschyzon merolae in Mammalian Cells

Background: Ascorbate peroxidase (APX) derived from Cyanidioschyzon merolae, a primitive red alga living in high temperature and acidic environments, has a greater anti-oxidative capacity than similar peroxidases occurring in other plants. In the present study, we examined the ability of Cyanidioschyzon merolae-derived APX (cAPX) to increase anti-oxidative capacity when expressed in mammalian cells. Materials and Methods: The cAPX gene was introduced into the mouse fibroblast-like cell line C3H10T1/2. Production of reactive oxygen species (ROS) and/or cell viability was assessed after heat, H2O2 and acid stimulation. Results: Heat and H2O2 stimulation caused ROS production. cAPX-expressing cells were more tolerant to oxidative stress induced by heat, H2O2 and acid stimulations than control cells lacking cAPX. Conclusion: Introduction of cAPX increases anti-oxidative capacity in mammalian cells

Crystal structure of Hop2-Mnd1 and mechanistic insights into its role in meiotic recombination

In meiotic DNA recombination, the Hop2-Mnd1 complex promotes Dmc1-mediated single-stranded DNA (ssDNA) invasion into homologous chromosomes to form a synaptic complex by a yet-unclear mechanism. Here, the crystal structure of Hop2-Mnd1 reveals that it forms a curved rod-like structure consisting of three leucine zippers and two kinked junctions. One end of the rod is linked to two juxtaposed winged-helix domains, and the other end is capped by extra ?-helices to form a helical bundle-like structure. Deletion analysis shows that the helical bundle-like structure is sufficient for interacting with the Dmc1-ssDNA nucleofilament, and molecular modeling suggests that the curved rod could be accommodated into the helical groove of the nucleofilament. Remarkably, the winged-helix domains are juxtaposed at fixed relative orientation, and their binding to DNA is likely to perturb the base pairing according to molecular simulations. These findings allow us to propose a model explaining how Hop2-Mnd1 juxtaposes Dmc1-bound ssDNA with distorted recipient double-stranded DNA and thus facilitates strand invasion

サイケツ ヲ ウケル 1サイ ノ コドモ ト オヤ ヘノ エンジョ ナイヨウ

背景 小児看護領域において, 子どもの権利条約の批准後, プレパレーションの概念が広まった. しかし, 小児専門病院の看護師に比べ, 総合病院の看護師のプレパレーションの認識は低く, プレパレーションの実践の普及がされていない実態がある. さらに, 採血を受ける子どもに関する研究は多いが, 自己の思考により物事をみることができない発達段階である2歳未満の年少の子どもを対象とした研究は少ない. 目的 採血を受ける1歳の子どもとその親への援助内容を明らかにする. 方法 全国の小児外来を標榜する総合病院131施設の外来に勤務する看護師のうち, 子どもの採血に関わる786名を対象に質問紙調査を行い, 自由記述の内容を分析しカテゴリー化した. 結果および考察 282名 (回収率35.8%) から回答が得られた. 看護師のプレパレーションの認知は, 知っている51.1%, 知らない45.0%であり先行研究より認識が高まっていた. しかし, 親の付き添いがないと回答した看護師は66.5%であり, さらに子どもの姿勢は寝かされ, 身体の抑制を受けながら採血を受けていたことが明らかとなった. 自由記述の内容分析では, 1歳の子どもと親への援助内容から看護師は, 子どもの発達段階に応じた援助ができていたが, 親が付き添う採血のケアモデル提示が少ないことが明らかとなった. 結論 看護師が, 1歳の子どもの発達段階に応じた援助を行うために, 子どもの権利およびプレパレーションの概念を学習する機会の提供が必要である. さらに, 親が付き添う採血のケアモデルの提示をする必要があることが示唆された.Background Nurses\u27 awareness of preparation at general hospitals is lower than that of nurses at specialist pediatric clinics, which has impeded the spread of preparation being put into practice throughout Japan. Although many studies have investigated children undergoing blood collection, few have examined young subjects aged less than two years.Objective This study aimed to clarify the state of blood collections conducted on one-year-old patients and support offered to parents.Methods A questionnaire survey targeting 786 outpatient nurses who routinely collected blood from pediatric patients at 131 general hospital facilities nationwide was conducted. Free comments were analyzed and content was categorized.Results/Discussion Answers were received from 282 nurses (recovery rate: 35.8%). It was found that nurses\u27 awareness of preparation had increased from that reported in previous studies to 51.1%. However, 66.5% of nurses reported that parents did not accompany children during blood collection. Analysis of free comments revealed that although nurses were offering parents support in accordance with children\u27s developmental stage, few care models incorporating parental attendance during blood collection had been presented.Conclusions Opportunities to study the concepts of child rights and preparation need to be provided so that nurses can give support in accordance with the developmental stage of one-year-old children undergoing blood collection. Results also suggested that a care model incorporating parental attendance during blood collection needs to be presented

カゾク ガ ツキソッタ バアイ ノ ヨウジ ノ サイケツ ニ タイスル タイショ コウドウ ノ カンサツ ブンセキ

背景 1994年に児童の権利に関する条約をわが国が批准してから, 小児看護領域では子どもの権利の保障としてプレパレーションの概念が広まった. 特に認知能力や言語的表現力が未熟な幼児に対して, 身体的苦痛が伴う検査や処置では子どもの理解力に応じたプレパレーションの実施が重要であることが明らかにされている. しかし検査や処置の場面構成は医療者が主体であるため, 採血場面に親の付き添いを除外する施設が多い. そのため親から分離された幼児は, 不安の増大により処置室への入室も困難な状態を引き起こす. 親が採血に付き添う場合に, 子どもがどのような反応や行動をとるかは明らかにされていない. 目的 家族が付き添う幼児後期の子どもの採血場面をビデオに録画し, 子どもと家族の相互の関係性から対処行動を明らかにする. 方法 小児外来で家族が付き添って採血を受ける幼児後期の子どもが, 採血を受けて退室するまでの行動をビデオに録画し, 採血終了後に家族へ半構成的面接を行った. 結果 採血前は【緊張の高まり】【誘導を受け入れる】【抵抗する】【自分なりの方法で心の準備をする】の4カテゴリーと8サブカテゴリー, 採血中は【誘導を受け入れる】【自分なりの方法で取り組む】【苦痛を表現する】【緊張をとく】の4カテゴリーと8サブカテゴリー, 採血後は【終了を行動する】【緊張がとける】【満足感を得る】の3カテゴリーと5サブカテゴリーが抽出された. 結論 採血場面に家族が付き添った幼児後期の子どもは, 家族が寄り添うことで安心を得ることができ, 調整能力を発揮し, 子どもは自分なりの方法で立ち向かうことができる. また採血中は痛みや苦痛をその場で受け止めてもらえる家族の存在により, 子どもは少しずつ緊張をとくことができる. さらに家族からの称賛により子どもは満足を得ることができ, 早期に日常の姿に戻ることができた.Background Japan ratified a treaty about the Child Rights in 1994, a general idea of preparation spread out as security of the rights of children in the child care region. For the infants that cognitive ability and verbal power of expression are immature, it is revealed that enforcement of preparation which accepted the understanding of children is important by testing and the treatment that physical pain is associated with. However, as for the scene constitution of testing and the treatment, there are many institutions excluding the attendant of the parent in the drawing blood scene because a medical person is the main constituent. Therefore the infants isolated from a parent cause a state having difficulty in admission to the dressing room by an increase of the anxiety. When a parent goes with drawing blood, it is uncertain what kind of response and behavior children take. Also, it is uncertain what kind of response and behavior children take when a parent goes with drawing blood.Purpose We determine coping behavior from the mutual relations of children and the family by shooting the drawing blood scene of the infants whom a family attends with a video camera.Methods We recorded behavior before undergoing drawing blood, and being discharged from the infants who underwent drawing blood with a family in pediatric outpatient department on a video .And had an interview to a family

ヨウジ ゼンキ ノ コドモ ガ ウケル サイケツ ニ ドウセキ スル ハハオヤ ノ ストレス

背景 近年,小児看護領域において急速にプレパレーションという概念が普及してきた.医療処置を受ける子どもにプレパレーションを提供し,その効果を最大に引き出すためには,子どもが最も信頼している母親の存在が重要である.しかし,我が国では,母親が同席し泣き叫ぶ子どもをみることで不安が増大する,また子どもは母親が助けてくれないことによって不審感をもつなどを理由に,同席を推奨する施設は少ない.目的 幼児前期の子どもの採血に同席した母親27名を対象に,唾液中アミラーゼ(AMY値),STAI(状態不安得点)の測定及び母親へのインタビューを行い,子どもが受ける採血に同席する母親のストレスを明らかにすることを本研究の目的とした.結果及び考察 AMY値と状態不安得点は採血前から10分後に有意に低下したが,採血前の状態不安得点は標準レベルであった.また,母親は採血に同席することについて【一緒が安心】と感じていた.子どもが不安や恐怖で泣くことがあっても,子どもの年齢や過去の経験から予測可能な子どもの反応であり,心の準備ができていたことから【泣くことの受容】に繋がったと考えられる.結論 子どもが受ける採血に同席した母親のAMY値と状態得点からは,これまで医療者の間で信じられてきた同席による母親の不安の増大は否定された.しかしながら,母親は採血時の姿勢について【抱っこの難しさ】も感じているため,単に同席を勧めるだけでなく,安全性の高い固定法や幼児期前期に有効なディストラクションを具体的に示し,母親が自信をもって採血に臨めるよう支援することが重要である.Background Mothers are the persons who are often the most trusted individual by their children and their presence is important when preparing children for medical treatment that can be invasive,painful or generally frightening.This concept is often referred to as "preparation" and is at the heart of caring for the parent and child; as both go through the procedure,one as the patient the other as comforter and safe trusted presence for the child.In Japan,however,few hospitals agree with the concept of preparation that believing mothers may become increasingly anxious as they see their children crying and children may mistrust their mothers for failing to help.This paper seeks to examine stress levels of mothers who are participating in a preparation procedure of drawing blood. Method 27 mothers of children aged 1-3 years who were present when blood samples were taken from their children,agreed to participate in the research project.Informed consent was given for this research by the participants A sample of saliva was taken pre and post procedure and examined for salivary amylase(AMY) levels which are stress indicators.An increase would indicate higher stress and a decrease would indicate lower stress.A State-Trait Anxiety Interview (STAI)was also performed per and post procedure a high score would indicate high anxiety.A low score would indicate low score. Findings Salivary AMY levels and STAI scores decreased significantly 10 min after the blood sampling compared with before,STAI scores before blood sampling were normal,and mothers also felt reassured if they were present while the blood was being drawn. Discussion Mothers felt reassured when they were present while the blood was being drawn.Even if children cried due to anxiety or fear,mothers considered this reaction predictable because of the child\u27s age and their own experience; because the mothers were prepared for it as a result,they were able to tolerate the crying remaining calm and giving reassurance to the child.These findings go against the widespread belief among medical professionals that mothers become more anxious if they are present during treatment.However,mothers also found it difficult to hold their children during the blood drawing,so in addition to recommending that they be present,physicians must assist mothers in facing blood sampling with confidence,by teaching them how to hold the child still in a safe manner and showing them effective ways of distracting infants