Yazarlar, teknik çalışmalar için Jamie Thuy kabul etmek istiyorum. UL1TR000130 (Anti Blokaj Sistemi), AA022448 (Anti Blokaj Sistemi), ve Eczacılık USC Okulu - Araştırma SC CTSI NIH / NCRR / NCATS hibe ile bu iş, kısmen, desteklenmiştir. Buna ek olarak, IVM içeren ve hızlı çözünen oral filmler nazik Kür İlaç (Oxnard, CA) tarafından sağlanmıştır.

açıklanan tüm prosedür Southern California Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Komitesi (IACUC) tarafından onaylanmıştır. 1. Hayvan Taşıma ve Alışma <ol><li> tanıtım <ol><li> fareler hayvancılık koşullara gelmesini ve deney tekniği ele izleyen ilaç uygulama prosedürleri sırasında stresi azaltmak için izin verin. Bu süre boyunca, vücut ağırlığı ve her bir farenin gıda alımı kayıt; arzu edildiği takdirde, daha sonra, ilaç toksisite (isteğe bağlı) değerlendirmek için bir karşılaştırma invazif olmayan noktası olarak kullanılabilir. Eski da kilo bazlı ilaç dozunu belirlemek için kullanılabilir. Not: Biraraya, alışma süresinden uzunluğu hayvan etkileşim deneyi sayısına göre belirlenmelidir. Bizim tecrübelerimize göre, C57BL / 6J suşu fareler sorun bizim tipik haftalık uzun alışma süresi içinde fazla 5 ~ den insanlar tarafından kullanımdan alıştırmak zorunda gözlenmiştir. , Mümkün isedeneyci (lar) da önceki raporlar gibi kokuyor gibi yeni uyaranlar, işaret olarak alışkanlık dönemi 11 engelleyebilir, hayvanlar dokunmadan önce kokulu losyonlar, sabunlar ve parfüm kullanmaktan kaçının teşvik edilmektedir. </li></ol></li><li> adımlar <ol><li> en az 5 gün önce Çalışmanın başlamasından itibaren, bir sonraki deneyler oluşacak olan odasına hayvanları aktarın. </li><li> sıkıca kuyruk ortasında tutarak ve yavaşça kafesin dışına ve ağırlık tekneye hayvan kaldırarak farenin ağırlığını kaydedin. <ol><li> O kuyruk 12 distal yönünü kavrama oluşabilecek deglove yaralanmaları önlemek için düzgün olarak, kuyruk medial / proksimal yönünü kavramak için akılda cagekeeping dışında kaldırdı kısa sürede ölçekte fare yerleştirin. </li><li> Fare güvenle tartmak tekne içine yerleştirilmiş edildikten sonra ağırlık değerini kaydedin. </li></ol></li><li> Bir RECOM için yiyecek ağırlığını kaydedindeney 13 sırasında yakalama potansiyel toksisite yardımcı olacak temel besin tüketim düzeylerini saptamak amacıyla, 2 gün üst üste iyileşiyorlardı. </li></ol></li></ol> Besleme Needle 2. Hazırlık, Sakaroz Çözüm ve Bireysel ODS <ol><li> deney için hazırlık paslanmaz çelik sonda iğneler sterilize etmek için bir otoklav kullanın. gerekirse 2 iğne / grubu ve 1 ek iğne hazırlayın back-up olarak kullanılır. Not: Bu çalışma, kavisli bir 1.5 inç, 18 g, kullanılır, iğne sonunda 2.25 mm&#39;lik top ile iğne besleme paslanmaz çeliktir. </li><li> (Ağırlık / hacim) sukroz çözümü, bir 4.25% oluşturmak için sakarozun 0,85 g 20 mL su ilave edin. Not: sükroz değerlendirilmesi için diğer düşük dozlarda altında parametrelerine bağlı olarak potansiyel olarak da 14 kullanılabilir. </li><li> Çapı 0.5 cm inç bir parçası haline standart dikdörtgen test şeridi keserek her hayvan için sözlü eritme şeridi (ODS) hazırlayın bizeticari olarak satılan bir tek çeyrek inç delik zımba ing. Bu tasvirde kullanılan dikdörtgen deney şeridi ivermektin 6 mg (IVM) ihtiva edecek şekilde formüle ve bu boyutuna kesme olduğunda, arzu edilen her bir hayvan için 10 mg / kg dozda yaklaşan her bir dairesel parça için 0.21 mg dozaj (eden tahsis edilmiş doz / hayvan). Bu doz / konsantre dikdörtgen şerit genel formülasyonun su seyreltilmesi ve / veya farklı bir boyutu şerit kesilerek ayarlanabilir. </li></ol> 3. İlaç - Teslim <ol><li> Daha önce adımlarda 1.1-1.2 açıklandığı gibi ağırlık değerlerini kaydedin. </li><li> kilo kaydedildikten sonra, metal ızgara kafes üstüne fareyi hareket ettirin. Bir kez orada, baskın başparmak, işaret parmağı, ve 3. parmağı kullanarak, yavaşça kuyruk çekin. Bu fare için yanlışlıkla kavrama tel kafes üst sebep ve uzak deneyci onun vücut germe, sırtını dışarı uzatmak olacaktır. </li><li> olmayan do ile boyun ense kavramakminant başparmak, işaret parmağı, ve 3. parmak rahat güvenli bir kavrama ile hayvan dizginlemek için. Not: Daha küçük el ile bir deneyci da arka ense geri çekin ve fare daha güvenli bir tutuş elde etmek için, aynı elin parmak inci 4 kullanabilirsiniz. Buna ek olarak, ayrı ayrı tercih, yüzük parmağı ve / veya serçe parmağı bağlı olarak, aynı zamanda kuyruk sabitlemek için kullanılabilir, ancak bu 12 gerekli değildir. </li><li> dik bir konumda fare tutun. </li><li> Dikkat: fare mukoza zarlarının renk dikkate alınmalıdır. Bir mor / mavi bir görünüm deneyci çok sıkı ense cinching ve fare nefes alamaz bir göstergesidir. <ol><li> Bu durum ortaya çıkarsa, derhal geri aşağı metal kafes üstüne fare koymak ve hayvan dizginlemek için bir denemede önce ~ 2-3 dk dinlenmeye bırakın. </li></ol></li><li> Bir tat yoğunluğu arttırıcı madde olarak hizmet% 4.25 sukroz çözeltisi içinde sonda iğne ampul ucu BatmakODS için yapıştırıcı. </li><li> oral eriterek şeridi eklemek için sonda iğne ampul ucu ile bir önceden kesilmiş ODS bastırın. </li><li> burun delikleri ve / veya ağız yakın yerleştirerek fare ODS sunmak ve hayvan filmi tüketmek için izin verir. Alternatif ODS fareler yutmak için dil üzerine doğrudan ince şerit koyarak ağız boşluğuna topik teslim edilebilir. </li><li> enfeksiyonun yayılmasını önlemek için ilaç dozaj oturumu sırasında iğne orta grup değiştirin. </li></ol>

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DD bu yönteme bir Co-Inventor olarak bir patent için başvurdu ve kronik hayvanlar üzerinde test (hızlı kullanma Laboratuvar Hayvanlarında İlaç aktif maddelerinin oral teslim için bu tekniğin iyi bilinen ve kolaylıkla kullanılabilir hale getirmek için bilimsel komite ile bu yöntemi paylaşımı ilgileniyor etti Ağız Filmler eriterek (Davies, Co-Inventor, Patent Başvurusu 6-23-2014 yayımlandı). Başka hiçbir rakip mali çıkarları belirtilmiştir.

Ağızdan sonda potansiyel tehlikeye veri toplama ve hayvan yıpratma 1 yüksek oranda neden sayısız komplikasyonlarla ilişkilidir. Burada, oral ilaç verilmesi basit bir yöntem olup, üzerinden fareler (ve potansiyel olarak diğer hayvanlar) oral olarak eritilmesi şeridin (ODS) ile tercih edilen bir ilaç tüketen kolayca ve güvenilir bir şekilde olabilir sokulur. Özellikle, bu yöntemin kullanılması, oral gavaj için güvenli, uygun ve insani bir alternatiftir. oral ga...

klinik geliştirme için klinik öncesi (hayvan testi) için bir ilacın başarılı bir çeviri için, bir dizi olan (akut ve kronik farmakolojik ve toksikolojik çalışmalar, amaçlanan ilaç verme klinik yol kullanılarak hayvan modellerinde gerçekleştirilmesi gerekir, iyi tanımlanmış normalde ağız yolu). Bu amacı gerçekleştirmek için, en çok klinik öncesi çalışmalar halen verilen bir oral gavaj prosedürünü kullanan hayvana, test bileşiğinin, bir tutarlı ve hassas bir teslimat Bu metod bu. Bununla birlikte, bir çok durumda, ağızdan sonda, hayvan tarafından tolere artan kanıtlar yöntem yük indüksiyon 1 önemli bir miktarı ile ilişkili olduğunu göstermektedir değildir. Bu gibi uzun vadeli klinik öncesi çalışmalar gerektiren kronik yaşam boyu durumlar için tedavi geliştirilmesinde de geçerlidir olduğu, ancak hemen hemen her nöropsikiyatrik bozukluğun, bunlarla sınırlı değildir. instance, Alzheimer hastalığının bir transgenik fare modelinde, son zamanlarda sonda yöntemi kendisi yüzünden stres 2 kaynaklı dozlama için deney sonuçlarını bulandırabilir kanısına varıldı. ve kendisini - - anlamlı deneysel bütünlüğünü tehlikeye noktasına alkol alımı düzeyini azaltabilir Benzer şekilde, alkol tüketiminin bir fare modelinde, kendi laboratuvarından yayınlanmamış bulgular sonda tüpünün tekrarlayan ekleme bulduk paradigma (Şekil 1). ağızdan sonda prosedürü ile ilişkili yukarıda belirtilen sınırlamaları göz önüne alındığında, önemli bir çaba klinik öncesi ağızdan ilaç verilmesinin yeni yöntemler geliştirmeye yönelik konulmuştur. Güncel alternatif yaklaşımlar Grubu Derneği&#39;nden bütün bunlar, su 7 ve gofret kraker 8 içme, fıstık ezmesi karışımları 3, jelatinimsi kalıplar 4-5, çikolata pelet 6 içine Test bileşimi içeren içeriryetersizlik gibi konularda değişen derecelerde ated hayvan modelinde güvenilir kullanılmak üzere ilaç daha az kolaylıkla 9 tüketilen yapan denek ya da lezzet için bireysel tercihiyle test bileşiği alımını kabul eksikliği. Bu tarifnamede sunulan prosedür istenen ilaç oral test hayvanına test bileşiği tatbik etmek için kullanılabilir kolaylıkla kolayca ve bir aromalı oral çözünen şeridin (ODS) dahil edildiği bir yöntem tarif eder. Bu kağıt fareler kullanılarak ODS yöntemin etkinliğini gösteren odaklanır. Ancak, aynı yöntem aynı zamanda diğer kemirgen yanı sıra büyük memeli konularda yararlı olmayacağını beklemek için hiçbir neden yoktur. Klinik, ODS kullanımının zorlukları 10 yutma üstesinden gelmek için bir yol olarak, pediatrik ve geriatrik hastalarda kabul edilmesi başlıyor. Biz ODS de güvenli, rahat bir şekilde klinik öncesi ilaç keşif programları başarılı bir şekilde kullanılabilir öneriyoruzağızdan sonda ile ve insancıl bir alternatif.

<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

Insanlarda yeni terapötiklerin test öncesinde kısa ve uzun vadeli klinik öncesi (yani, hayvan), tekrarlayan farmakolojik ve toksikolojik test gereklidir. Çoğu durumda, tercih edilen uygulama yolu ağızdan uygulama yoluyladır. Şu an için, oral uygulama çoğunlukla hayvan modelinde eden tutarlı ve kesin dozaj elde çünkü, kısmen, bir oral gavaj prosedür kullanılarak gerçekleştirilir. Bu yöntem iyi kurulmuş olmasına rağmen hayvan yıpratma yüksek oranda neden olabilir komplikasyonları var. Bu amaçla, burada kişiye prosedürü sadece daha sonra hızlı bir şekilde içine alınır test hayvanına sunulabilir istenen ilaç oral şeridin (ODS) eritilmesi, bir tat yoğunluğu arttırıcı madde olarak dahil edilen, oral gavaj metoduna bir alternatif tarif denek minimal manipülasyon. Bu yazıda, ODS yöntemi kullanarak preklinik, oral ilaç teslimat, güvenli, rahat ve insancıl alte temsil ettiğini göstermektediroral gavaj ile rnative.

Aşağıdaki temsili incelemelerde, sosyal içme önce 15 açıklandığı gibi bir 24 saat iki şişe seçim (TBC) paradigma kullanılarak modellenmiştir. Kısaca, fareler su içeren biri çözeltisi iki şişe, erişebildiği ve diğer bir% 10 (hac / hac) etanol çözeltisi. Ortalama 10E (% 10 etanol /% 90 su hac / hac) emme gruplarında benzer şekilde Konular daha sonra, ilaç tedavi gruplarına ayrıldılar. S...

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

Aşağıdaki protokol daha yaygın olarak kullanılan oral gavaj yönteminin yerine bir oral eritilmesi şeridi (ODS) ile kronik oral ilaç uygulaması için yeni bir yöntem tarif eder. Biz ODS yöntemi kullanarak preklinik oral ilaç dağıtım ağızdan sonda güvenli, rahat ve insancıl bir alternatif temsil ettiğini göstermektedir.

Farmakolojik ve Toksikoloji Araştırmaları bir Oral Çözme Strip kullanmak: Hayvanlar için ağızdan sonda ile Basit ve İnsanca Alternatif

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

utilizing-an-orally-dissolving-strip-for-pharmacological

Research

JoVE Journal

Medicine

12.6K Görüntüleme.  University of Southern California School of Pharmacy.  Aşağıdaki protokol daha yaygın olarak kullanılan oral gavaj yönteminin yerine bir oral eritilmesi şeridi (ODS) ile kronik oral ilaç uygulaması için yeni bir yöntem tarif eder. Biz ODS yöntemi kullanarak preklinik oral ilaç dağıtım ağızdan sonda güvenli, rahat ve insancıl bir alternatif temsil ettiğini göstermektedir. 

Video: 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.

Oral Skuamöz Hücre Karsinomu Ortotopik Fare Modeli Tümör Hücre Invasion Multi-foton Görüntüleme

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

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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.

Basit Fare İntrakranial Xenograftlarında Çalışmaları Kılavuzu Vida Yöntemi

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.

Alternatifi olarak dilaltı immünoterapi Akut Solunum Enfeksiyonları Karşı Koruma tetikleyebilecek

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.

Doku Dejenerasyonu / Yenilenme ve Kanser Araştırmaları Murin İndüklenebilir Telomeraz allelleri kullanılması

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.

Basit Aygıt Hızla Fare Bağırsak Tüm Mounts Hazırlanır

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.

Bir rekombinant adeno-ilişkili viral vektörün stereotaktik enjeksiyon yoluyla Parkinson Hastalığı için bir alfa-sinüklein Bazlı Fare Modeli Geliştirme

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.

Köpek arter ve venlerin Primer Endotel Hücreleri İzolasyon ve Kültür

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.

Sağlıklı çocuklarda Microbiome analiz için sözlü biyofilm örnekleme

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.

Yaşlı Yetişkinler için Lay Personel tarafından Ağız Sağlığı Değerlendirmesi

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.

Ağız Dışı ve Ağız İçi Dijital Fotoğrafçılığa Standartlaştırılmış Bir Yaklaşım

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