著者は、技術的な仕事のためのジェイミー・トゥイを承認したいと思います。この作業は、部分的には、研究によって、サポートされていたSC CTSI NIH / NCRR / NCATSを付与 - UL1TR000130（DLD）、AA022448（DLD）、および薬局のUSCの学校。また、IVMを含む速溶性経口フィルムは、親切にキュア製薬（オックスナード、CA）によって提供されました。

記載されている全ての手順は、南カリフォルニア大学の施設内動物管理使用委員会（IACUC）によって承認されています。 1.動物の取り扱いと馴化<ol><li>導入<ol><li>マウスはその後の薬物投与手順中にストレスを減少させるために飼育条件と実験の技術を扱うに順応することを可能にします。この時間の間、各マウスの体重および食物摂取量を記録します。所望であれば、それらは後に（オプション）薬物毒性を評価するために、比較の非侵襲的な点として使用することができます。前者はまた、重量ベースの薬物投与を決定することができます。 注：ちなみに、馴化期間の長さは、動物と相互作用する実験者の数に基づいて決定されるべきです。我々の経験では、C57BL / 6J系統のマウスは、私たちの典型的な一週間の馴化期間内より5〜よりも人によって取り扱いにhabituating悩みを持っていることが観察されています。実現可能な場合は、実験者は（複数可）も以前の報告は、このような臭いなどの新しい刺激を示すよう馴化期間11を妨げる可能性が、動物を取り扱う前に香りのローション、石鹸、香水の使用を控えることが奨励されます。 </li></ol></li><li>ステップ<ol><li>前研究の開始に少なくとも5日から開始し、その後の実験が起こる部屋に動物を転送します。 </li><li>しっかりと尾の真ん中をつかみ、静かにケージから出て、重量ボートに動物を持ち上げることにより、マウスの重量を記録します。 <ol><li>尾12の遠位面を把握するときに発生することができますdegloving怪我を避けるために適切に、尾の内側/近位側面を把握するために念頭に置いてcagekeepingから持ち上げされるとすぐにスケール上にマウスを置きます。 </li><li>マウスが安全に計量ボート内に配置された後に重量値を記録します。 </li></ol></li><li> RECOMのための食品の重量を記録します実験13の間に潜在的な毒性をキャプチャするのに役立ちますベースライン食料消費水準を確立するために、2日間連続直し。 </li></ol></li></ol>摂食ニードルの作製、ショ糖液、および個々のODS <ol><li>実験の準備のためにステンレス鋼の強制飼養針を滅菌するオートクレーブを使用してください。 2針/グループとすることが必要な場合はバックアップとして使用する1追加の針を準備します。 注：この研究は、1.5インチ、針の端で2.25ミリメートルボールと湾曲し、18 G、ステンレス鋼供給針を利用しました。 </li><li> （w / v）のスクロース溶液を4.25％に作成するために、スクロース0.85gのに水20mlを加えます。 注：ショ糖の評価の他の低用量の下のパラメータに応じて、潜在的にも14を使用することができます。 </li><li>直径0.5センチメートルインチ片に標準の長方形の試験片を切断して各動物の経口溶解ストリップ（ODS）を準備たち市販の単四分の一インチの穴パンチャーをる。この実演で使用長方形の試験片は、イベルメクチン6mgの（IVM）を含有するように配合し、この大きさに切断した場合、所望の各動物について10mg / kgの用量に近似し、各個々の円形片に0.21 mgの用量を（私達の割り当てられました用量/動物）。この用量は、短冊の全体的な製剤を希釈及び/又は異なる大きさにストリップを切断/濃縮することによって調整することができます。 </li></ol> 3.薬剤 - 配達<ol><li>以前のステップ1.1から1.2で説明したように重み値を記録します。 </li><li>体重を記録した後、金属グリッドケージ上部にマウスを移動します。そこから、支配的な親指、人差し指、および3 番目の指を使用して、そっと尾に引き戻します。これは、不注意グリップにマウスワイヤーケージのトップ原因と離れ実験からその体を伸ばし、その背中を伸ばします。 </li><li>非DOと首筋をつかん快適に安全であるグリップで動物を抑制するminant親指、人差し指、および3 番目の指。 注：小さい手で実験者はまた、背中の首筋を引き戻すと、マウスのより安全なホールドを取得するために、同じ手の指番目の 4を使用することができます。さらに、個々の嗜好、薬指、および/ ​​または小指の指によって尾を固定するためにも使用され得るが、これは12必要ではありません。 </li><li>直立位置にマウスを置きます。 </li><li>注意：マウスの​​粘膜の色を観察します。紫/青外観は、実験者があまりにもきつく首筋を締めるされ、マウスが息ができないことを示しています。 <ol><li>この問題が発生した場合は、すぐに戻ってダウン金属ケージの上にマウスを置くと、それは動物を拘束するために再試行する前に〜のために2〜3分を休ませることができます。 </li></ol></li><li>味物質として機能するように4.25パーセントのショ糖溶液中に強制経口投与針の電球の先端を水没ODSへの接着剤。 </li><li>経口溶解ストリップを取り付けるために強制経口投与針の電球の先端とのプレカットODSを押し下げます。 </li><li>鼻孔および/または口の近くに配置することにより、マウスにODSを提示し、そして動物がフィルムを消費することができます。あるいは、ODSは、飲み込むことがマウスについての舌の上に直接薄いストリップを配置することにより、口腔内に局所的に送達することができます。 </li><li>感染の拡大を防止するために、薬物投与セッション中に針半ばグループを交換してください。 </li></ol>

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DDは 、この方法での共同発明者として特許を申請し、慢性動物実験（ ファストを使用した実験動物における薬物活性物質の経口送達のためのこの技術はよく知られており、容易に利用できるようにする科学委員会でこの方法を共有することに興味ありました経口フィルムを溶解させる （Davies氏は、共同発明者、特許出願は、2014年6月23日出願しました）。 他の競合する金融持分は指摘されていません。

強制経口投与は、潜在的に危険にさらさデータ収集と動物のスレ1の高い速度をもたらす数々の合併症に関連付けられています。ここでは経口薬物送達の簡単な方法は、（他の動物と潜在的に）どのマウスを介して導入され、容易かつ確実に経口溶解ストリップ（ODS）を介して選択される薬物を消費することができます。注目すべきことに、この方法の使用は、強制経口投与のために?...

前臨床（動物試験）の臨床開発の薬剤の成功翻訳、明確に定義された一連のためには、急性および慢性の薬理学的及び毒物学的研究がある（投与の意図される臨床経路を用いて動物モデルで実行される必要があります通常、経口経路）。この目標を達成するために、ほとんどの前臨床試験は、現在指定された経口胃管栄養法の手順を利用し、動物に試験化合物の一貫した正確な送達におけるこの方法は、結果は。しかし、多くの場合、経口胃管栄養法はよく動物により許容され、蓄積の証拠は、この方法は、ストレス誘導1のかなりの量に関連していることを示唆していません。 これは、このような場合であるが、ほぼすべての神経精神障害、これらに限定されないなどの長期的な前臨床研究を、必要とする慢性の生涯条件のための治療法の開発に特にそうです。イン用ンスは、アルツハイマー病のトランスジェニックマウスモデルでは、最近胃管栄養法自体は、投薬誘発性ストレス2による実験結果を混乱させる可能性が示唆されました。それ自体は - - 大幅実験の整合性を危うくする点にアルコール摂取のレベルを減らすことができます。同様に、アルコール消費のマウスモデルでは、私たち自身の研究室からの未発表の所見は、胃管栄養チューブの繰り返し挿入があることを見出しましたパラダイム（ 図1参照）。 強制経口投与手順に関連する前述の限界を考えると、かなりの努力が前臨床経口薬物送達の新規な方法を開発に向けて置かれています。現在の別のアプローチはassociあるすべてが、水7、ウエハクラッカー8飲料 、ピーナッツバターの混合物3、ゼラチン状の金型4-5、チョコレートペレット6に試験化合物を組み込むことが含まれます動物モデルで確実に使用することができないこと、試験化合物の被験者による取り込みまたは薬物が少なく容易に9を消費させる風味のための個々の好みの採用の欠如などの問題の様々な程度でated。 本明細書に導入された手順は、簡単かつ容易に経口で試験動物に試験化合物を投与するために使用することができる所望の薬剤を風味の経口溶解ストリップ（ODS）に組み込まれた方法を記載します。本稿では、マウスを使用してODS手法の有効性を実証するに焦点を当てています。しかし、同法はまた、他の齧歯類と同様に、より大きな哺乳動物被験体に有用ではないであろうことを期待する理由はありません。臨床的には、ODSの使用が困難10を嚥下克服する方法として、小児及び老人患者で採用され始めています。私たちは、ODSも安心、便利なように前臨床創薬プログラムで正常に使用できることを提案します強制経口投与であり、人道的な代替。

<table border="0" cellpadding="0" cellspacing="0" width="948">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:203px;">Orally Dissolving Strip&#160;</td>
			<td style="width:185px;">Cure Pharmaceutical</td>
			<td style="width:89px;"></td>
			<td style="width:471px;">Available upon special request. Alternatively, ODS may also be prepared in-house.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:203px;">Gavage Feeding Needle</td>
			<td style="width:185px;">VWR</td>
			<td align="right" style="width:89px;">7912</td>
			<td>Our rodent studies utilized a 18-gauge feeding needle. This size may need to be adjusted for use in other mammilian species.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:203px;">Sucrose</td>
			<td style="width:185px;">Sigma Aldrich</td>
			<td style="width:89px;">S0389-500G</td>
			<td></td>
		</tr>
	</tbody>
</table>

utilizing an orally dissolving strip for pharmacological and toxicological studies: a simple and humane alternative to oral gavage for animals

ヒトにおける新規治療薬をテストする前に短期及び長期前臨床（ すなわち 、動物）、反復的な薬理学的および毒物学的テストが必要です。ほとんどの場合、好ましい投与経路は、経口送達を介するものです。現時点では、経口送達は、主に、それが動物モデルで一貫して正確な投与を達成することができるので、部分的には、経口胃管栄養法の手順を使用して達成されます。この方法は十分に確立されているが、それは動物の損耗率の高さになることが合併症を持っていません。この目的のために、ここで紹介する手順は、単にそれがその後急速に取り、試験動物に提示することができる所望の薬剤を経口ストリップ（ODS）を溶解、味物質に組み込まれた強制経口投与法に代わるものを説明します被験者の最小限の操作。ここで、我々は、ODS法を用いた前臨床、経口薬物送達は、安全で便利、そして人道的なアルテを表すことを示しています強制経口投与にrnative。

以前15に記載されているように 、以下の代表的な研究では、社会的な飲酒は、24時間の二瓶選択（TBC）パラダイムを用いてモデル化しました。簡潔には、マウスは、水を含有一つは溶液の2つのボトルへのアクセスを有し、他の10％（v / v）のエタノール溶液。平均10E（10％エタノール/ 90％水v / v）の摂取がグループ間で類似していたように、被?...

Watch this Scientific Journal Video about Utilizing an Orally Dissolving Strip for Pharmacological and Toxicological Studies: A Simple and Humane Alternative to Oral Gavage for Animals at JoVE.com

Utilizing an Orally Dissolving Strip for Pharmacological and Toxicological Studies: A Simple and Humane Alternative to Oral Gavage for Animals

以下のプロトコルは、より一般的に使用される経口栄養法の代わりに経口溶解ストリップ（ODS）を使用して、慢性経口薬物投与のための新規な方法を説明します。私たちは、ODS法を用いた前臨床、経口薬物送達は、経口栄養に、安全で便利な、そして人間的な代替手段を表すことを示しています。

薬理学的および毒性試験のための経口溶解ストリップを利用：動物のための強制経口投与にシンプルで動物愛護の代替を

Prior to testing novel therapeutics in humans, short and long term preclinical (i.e., animal), repetitive pharmacological and toxicological testing is ...

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12.6K ビュー.  University of Southern California School of Pharmacy. 以下のプロトコルは、より一般的に使用される経口栄養法の代わりに経口溶解ストリップ（ODS）を使用して、慢性経口薬物投与のための新規な方法を説明します。私たちは、ODS法を用いた前臨床、経口薬物送達は、経口栄養に、安全で便利な、そして人間的な代替手段を表すことを示しています。 

ビデオ: Utilizing an Orally Dissolving Strip for Pharmacological and Toxicological Studies: A Simple and Humane Alternative to Oral Gavage for Animals

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

口腔扁平上皮癌の同所性マウスモデルにおける腫瘍細胞の浸潤の多光子イメージング

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

医学

A Simple Guide Screw Method for Intracranial Xenograft Studies in Mice

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma (glioma) and medulloblastoma. The widely used stereotactic approach only allows for the injection of a single animal at a time, is labor intensive and requires highly specialized equipment. The guide screw method, initially developed by Lal et al.,1 was developed to eliminate cumbersome stereotactic procedures. We now describe a modified guide screw approach that is rapid and exceptionally safe; both of which are critical ethical considerations. Notably, our procedure now incorporates an infusion pump that allows up to 10 animals to be simultaneously injected with tumor cells.
To demonstrate the utility of this procedure, we established human U87MG glioma cells as intracranial xenografts in mice, which were then treated with AMG102; a fully human antibody directed to HGF/scatter factor currently undergoing clinical evaluation2-5. Systemic injection of AMG102 significantly prolonged the survival of all mice with intracranial U87MG xenografts and resulted in a number of complete cures. 
This study demonstrates that the guide screw method is an inexpensive, highly reproducible approach for establishing intracranial xenografts. Furthermore, it provides a relevant physiological model for validating novel therapeutic strategies for the treatment of brain cancers.

In order to evaluate novel therapeutic paradigms for the treatment of glioma, physiological relevant models are essential. We utilize an implantable guide screw procedure for establishment of intracranial xenograft models that is more rapid and safer than stereotactic approaches.

マウスにおける頭蓋内異種移植研究のための簡単​​なガイドスクリュー方式

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma ...

Sublingual Immunotherapy as an Alternative to Induce Protection Against Acute Respiratory Infections

Sublingual route has been widely used to deliver small molecules into the bloodstream and to modulate the immune response at different sites. It has been shown to effectively induce humoral and cellular responses at systemic and mucosal sites, namely the lungs and urogenital tract. Sublingual vaccination can promote protection against infections at the lower and upper respiratory tract; it can also promote tolerance to allergens and ameliorate asthma symptoms. Modulation of lung&#8217;s immune response by sublingual immunotherapy (SLIT) is safer than direct administration of formulations by intranasal route because it does not require delivery of potentially harmful molecules directly into the airways. In contrast to intranasal delivery, side effects involving brain toxicity or facial paralysis are not promoted by SLIT. The immune mechanisms underlying SLIT remain elusive and its use for the treatment of acute lung infections has not yet been explored. Thus, development of appropriate animal models of SLIT is needed to further explore its potential advantages. 
This work shows how to perform sublingual administration of therapeutic agents in mice to evaluate their ability to protect against acute pneumococcal pneumonia. Technical aspects of mouse handling during sublingual inoculation, precise identification of sublingual mucosa, draining lymph nodes and isolation of tissues, bronchoalveolar lavage and lungs are illustrated. Protocols for single cell suspension preparation for FACS analysis are described in detail. Other downstream applications for the analysis of the immune response are discussed. Technical aspects of the preparation of Streptococcus pneumoniae inoculum and intranasal challenge of mice are also explained.
SLIT is a simple technique that allows screening of candidate molecules to modulate lungs&#8217; immune response. Parameters affecting the success of SLIT are related to molecular size, susceptibility to degradation and stability of highly concentrated formulations.

The present work illustrates the convenience of using sublingual immunotherapy to boost the innate immune response in the lungs and confer protection against acute pneumococcal pneumonia in mouse.

代替として舌下免疫療法は、急性呼吸器感染症に対する保護を誘導するために

Sublingual route has been widely used to deliver small molecules into the bloodstream and to modulate the immune response at different sites. It has been ...

Utilizing Murine Inducible Telomerase Alleles in the Studies of Tissue Degeneration/Regeneration and Cancer

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that cause tissue degeneration during ageing. Cancer, with incidence rates greatly increasing with age, is characterized by short telomere lengths and high telomerase activity. To study the roles of telomere dysfunction and telomerase reactivation in ageing and cancer, the protocol shows how to generate two murine inducible telomerase knock-in alleles 4-Hydroxytamoxifen (4-OHT)-inducible TERT-Estrogen Receptor (mTERT-ER) and Lox-Stopper-LoxTERT (LSL-mTERT). The protocol describes the procedures to induce telomere dysfunction and reactivate telomerase activity in mTERT-ER and LSL-mTERT mice in vivo. The representative data show that reactivation of telomerase activity can ameliorate the tissue degenerative phenotypes induced by telomere dysfunction. In order to determine the impact of telomerase reactivation on tumorigenesis, we generated prostate tumor model G4 PB-Cre4 PtenL/L p53L/L LSL-mTERTL/L and thymic T-cell lymphoma model G4 Atm-/- mTERTER/ER. The representative data show that telomerase reactivation in the backdrop of genomic instability induced by telomere dysfunction can greatly enhance tumorigenesis. The protocol also describes the procedures used to isolate neural stem cells (NSCs) from mTERT-ER and LSL-mTERT mice and reactivate telomerase activity in NSCs in vitro. The representative data show that reactivation of telomerase can enhance the self-renewal capability and neurogenesis in vitro. Finally, the protocol describes the procedures for performing telomere FISH (Fluorescence In Situ Hybridization) on both mouse FFPE (Formalin Fixed and Paraffin Embedded) brain tissues and metaphase chromosomes of cultured cells.

Telomere and telomerase play essential roles in ageing and tumorigenesis. The goal of this protocol is to show how to generate two murine inducible telomerase knock-in alleles and how to utilize them in the studies of tissue degeneration/regeneration and cancer.

組織の変性/再生と癌の研究におけるマウス誘導テロメラーゼ対立遺伝子を利用

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that ...

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe the use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations of superior and uniform quality to that which can be achieved by hand. The device comprises a base that holds 4 stainless steel rods and a top, which acts a cutting guide. The rods are inserted into the lumen of the small intestine [divided into thirds] and the colon. The rods and samples are then placed over a piece of filter paper or card into the holding slots in the base of the device. The top of the device is then positioned and serves as a cutting guide. The two angled sections in the center of the top piece are used to guide a knife or scalpel and cut the intestines longitudinally on the top of the rods. Once the intestinal sections have been cut, the top is removed and the card,&#160;tissue and rods gently removed from the device and placed on the bench. The rods are then gently rolled sideways to flatten and stick the intestinal segments onto the underlying piece of filter paper or card. The final preparation can then be examined or fixed and stored for later analysis. The preparations are invaluable for the study of intestinal changes in normal or genetically modified mouse models. The preparations have been used for the study and quantification of the effects of inflammation (colitis), damage, pre-cancerous lesions (aberrant crypt foci (ACFs) and mucin depleted foci (MDFs)) and polyps or tumors.

The use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations is described.

急速にマウス腸のホールマウントを準備するための簡単​​な装置

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

Development of an Alpha-synuclein Based Rat Model for Parkinson's Disease via Stereotactic Injection of a Recombinant Adeno-associated Viral Vector

In order to study the molecular pathways of Parkinson&#39;s disease (PD) and to develop novel therapeutic strategies, scientific investigators rely on animal models. The identification of PD-associated genes has led to the development of genetic PD models. Most transgenic &#945;-SYN mouse models develop gradual &#945;-SYN pathology but fail to display clear dopaminergic cell loss and dopamine-dependent behavioral deficits. This hurdle was overcome by direct targeting of the substantia nigra with viral vectors overexpressing PD-associated genes. Local gene delivery using viral vectors provides an attractive way to express transgenes in the central nervous system. Specific brain regions can be targeted (e.g. the substantia nigra), expression can be induced in the adult setting and high expression levels can be achieved. Further, different vector systems based on various viruses can be used. The protocol outlines all crucial steps to perform a viral vector injection in the substantia nigra of the rat to develop a viral vector-based alpha-synuclein animal model for Parkinson&#39;s disease.

This manuscript describes how viral vector-mediated local gene delivery provides an attractive way to express transgenes in the central nervous system. The protocol outlines all crucial steps to perform a viral vector injection in the substantia nigra of the rat to develop a viral vector-based animal model for Parkinson's disease.

組換えアデノ随伴ウイルスベクターの定位注射を介して、パーキンソン病のα-シヌクレインベースのラットモデルの開発

In order to study the molecular pathways of Parkinson&#39;s disease (PD) and to develop novel therapeutic strategies, scientific investigators rely on ...

Isolation and Culture of Primary Endothelial Cells from Canine Arteries and Veins

Cardiovascular disease is studied in both human and veterinary medicine. Endothelial cells have been used extensively as an in vitro model to study vasculogenesis, (tumor) angiogenesis, and atherosclerosis. The current standard for in vitro research on human endothelial cells (ECs) is the use of Human Umbilical Vein Endothelial Cells (HUVECs) and Human Umbilical Artery Endothelial Cells (HUAECs). For canine endothelial research, only one cell line (CnAOEC) is available, which is derived from canine aortic endothelium. Although currently not completely understood, there is a difference between ECs originating from either arteries or veins. For a more direct approach to in vitro functionality studies on ECs, we describe a new method for isolating Canine Primary Endothelial Cells (CaPECs) from a variety of vessels. This technique reduces the chance of contamination with fast-growing cells such as fibroblasts and smooth muscle cells, a problem that is common in standard isolation methods such as flushing the vessel with enzymatic solutions or mincing the vessel prior to digestion of the tissue containing all cells. The technique we describe was optimized for the canine model, but can easily be utilized in other species such as human.

Novel isolation methods of primary endothelial cells from blood vessels are needed. This protocol describes a new technique that completely inverts blood vessels of interest, exposing only the endothelial side to enzymatic digestion. The resulting pure endothelial cell culture can be used to study cardiovascular diseases, disease modelling, and angiogenesis.

犬の動脈と静脈からの初代内皮細胞の単離および培養

Cardiovascular disease is studied in both human and veterinary medicine. Endothelial cells have been used extensively as an in vitro model to study ...

Oral Biofilm Sampling for Microbiome Analysis in Healthy Children

Oral biofilm and its molecular analysis provide a basis for investigating various dental research and clinical questions. Knowledge of biofilm composition leads to a better understanding of cariogenic and periopathogenic mechanisms. Microbial changes taking place in the oral cavity during childhood are of interest for several reasons. The evolution of the child oral microbiota and shifts in its composition need to be analyzed further to understand and possibly prevent the onset of disease. At the same time, advanced knowledge of the natural composition of oral biofilm is needed. Early stages of caries-free permanent dentition with healthy gums provide a widely unaffected subgingival habitat that can serve as an in situ baseline for studying features of oral health and disease. Analysis of children's oral biofilm during different stages in life is thus an important theme in the field. Modern molecular analysis methods can provide comprehensive information about the bacterial diversity of such biofilms. To enable microbiota data comparison, it is important to standardize each step in the procedure for molecular data generation. This procedure spans from clinical sampling, Next Generation Sequencing (NGS), bioinformatic data processing, to taxonomic interpretation. One of the most critical factors here is biofilm sampling. Sampling in children is even more challenging in particular due to limited space in subgingival areas. We thus focus on the use of paper points for subgingival sampling. This article provides a detailed protocol for oral biofilm sampling of the subgingival sulcus, the mucosa, and saliva in children.

Changes in the oral microbiome throughout childhood are of growing interest. Comparison of different microbiome studies reveals a lack of standardized sampling protocols. Limited space makes sampling the sound subgingival sulcus of children challenging. Paper point sampling is presented here in detail as the method of choice for this area.

マイクロバイにおける健常児の口腔バイオ フィルム サンプリング

Oral biofilm and its molecular analysis provide a basis for investigating various dental research and clinical questions. Knowledge of biofilm composition ...

Oral Health Assessment by Lay Personnel for Older Adults

Oral health is an often-undervalued contributor to overall health. The literature, however, underscores the myriad of systemic diseases influenced by oral health, including type II diabetes, heart disease, and atherosclerosis. Thus, assessments of oral health, called oral screenings, have a significant role in assessing risk of disease, managing disease, and even improving disease by oral care. Here we present a method to assess oral health quickly and consistently across time. The protocol is simple enough for non-oral health professionals such as students, family, and caregivers. Useful for any age of patient, the method is particularly key for older individuals who are often at risk of inflammation and chronic disease. Components of the method include existing oral health assessment scales and inventories, which are combined to produce a comprehensive assessment of oral health. Thus, oral characteristics assessed include intraoral and extraoral structures, soft and hard tissues, natural and artificial teeth, plaque, oral functions such as swallowing, and the impact this oral health status has on the patient&#39;s quality of life. Advantages of this method include its inclusion of measures and perceptions of both the observer and patient, and its ability to track changes in oral health over time. Results acquired are quantitative totals of questionnaire and oral screening items, which can be summed for an oral health status score. The scores of successive oral screenings can be used to track the progression of oral health across time and guide recommendations for both oral and overall health care.

The goal of this protocol is to enable non-dental professionals to assess oral health status for research or health-screening purposes. Aspects assessed include lips, tongue, soft and hard tissues, natural and artificial teeth, oral cleanliness, plaque, swallowing, and impact of oral health on quality of life.

高齢者のレイ・スタッフによる口腔健康評価

Oral health is an often-undervalued contributor to overall health. The literature, however, underscores the myriad of systemic diseases influenced by oral ...

A Standardized Approach to Extra-Oral and Intra-Oral Digital Photography

Contemporary dentistry mandates a more comprehensive and personalized analysis of each patient. Technological advances in digital photography have played vital roles in diagnostic accuracy, treatment planning, execution of therapies, and outcome evaluations, including esthetic enhancement. Digital photography also provides an excellent platform for patient education, communication, and co-management of cases with other healthcare providers. However, intra-oral photography often faces challenges such as inaccessibility of areas to be captured, different moveable vs. fixed tissues involved, contamination with saliva or blood, and differing illumination needs on various locations. Thus, a more standardized and systematic approach is proposed for intra- and extra-oral documentation via digital photography to overcome the existing technical challenges. The current work will outline the appropriate equipment specifications (camera bodies, macro lens, and flashes), positions and postures of the operator and patients, proper techniques of tissue retraction, the use of appropriate intra-oral mirrors, and the essential elements such as aperture settings (F-stop), ISO, shutter speed, and white balance. This article aims to provide all dental professionals with an approachable linear array of guidelines to produce simplified and standardized visual tools for more efficient and effective documentation.

The present protocol describes a method to capture intra- and extra-oral photographs for digital documentation in the clinical practice of dentistry. The protocol highlights all the critical components involved and the commonly utilized settings to capture dental photographs for clinical documentation.

口腔外および口腔内デジタル写真への標準化されたアプローチ

Contemporary dentistry mandates a more comprehensive and personalized analysis of each patient. Technological advances in digital photography have played ...