The authors would like to thank John Kristensen and S&#248;ren Braad Andersen from the Department of Communication, Aarhus University Hospital, Denmark, for their exceptional help with filming and editing. Animal technician Diana Gyldenl&#248;ve and veterinarian Birgitte Kousgaard, Institute of Clinical Medicine Aarhus University Hospital, Denmark for assisting with animal care. This study was supported by the Lundbeck Foundation, the Laerdal Foundation for Acute Medicine, Central Denmark Region&#8217;s Research Foundation, Augustinus Foundation, Aase and Ejnar Danielsens Foundation, the Institute of Clinical Medicine Aarhus University Hospital, Br&#248;drene Hartmanns Foundation, Karen Elise Jensens Foundation, Fonden til L&#230;gevidenskabens Fremme, and Marie Dorthea og Holger From, Haderslevs Fond.

<ol>
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الكتاب ليس لديهم ما يكشف.

نظرا لتعقيده، لا يمكن إلا أن تنفيذ نموذج موضح في المرافق المعتمدة وذوي الخبرة في مجال البحوث الحيوانية. يجب الحصول على موافقة من قبل لجان الأخلاق المحلية قبل الشروع في التجارب، ويجب ضمان الرفق بالحيوان الأمثل في جميع الأوقات. كما يستند هذا النموذج على البقاء على قيد ...

الاختناق حول الولادة هو حالة حادة وغير متوقعة في كثير من الأحيان يرتبط بنقص التأكسج اعتلال الدماغ الإقفاري (عجل). الهدف العام من هذا البروتوكول هو لإظهار نموذج بقاء خنزير صغير من فترة ما حول الولادة التأكسج نقص تروية الدماغ. هذا النموذج يمكن أن تستخدم لدراسة تأثير درجات مختلفة من نقص الأكسجة نقص التروية على الدماغ حديثي الولادة والعلاجات التجريبية على الأعصاب، والتصوير بالرنين المغناطيسي والتحليل الطيفي (MRI وMRS) والمؤشرات الحيوية في سوائل الجسم مثل الدم والسائل النخاعي والبول . وقد أثبت هذا النموذج مفيد أيضا للتحقيق في نظام القلب والأوعية الدموية والجهاز التنفسي والكلى والكبد، والتي تتأثر في عالمي نقص الأكسجة نقص التروية. الاختناق حول الولادة هو نتيجة للخطر أثناء الوضع امدادات الاوكسجين أو في فترة ما بعد الولادة مباشرة. حساب المخاض الأحداث ميتة ل0660000 شخص حول العالم سنويا، حوالي ربع سكان العالم البالغ عددهم 2.9 طاحونةوقدرت وفيات الولدان أيون في عام 2012 3. وفي عام 2010 1.15 مليون طفل قد وضعت اعتلال الدماغ عند الوليد بعد الاختناق أثناء الولادة 4. HIE يعرف بأنه التهاب الدماغ في الأطفال الذين يولدون بعد 34 أسبوعا من الحمل يحدث في 1-3 / 1000 ولادة حية 5 في العالم الصناعي، و 8.5 / 1000 ولادة حية في البلدان النامية (4). خطر الموت هو 10-60٪، وخطر الإعاقة العصبية عند الناجين 30-100٪ 6،7. وتعزى 50200000 العجز تعديل الحياة سنة (سنوات العمر) للأحداث ميتة الولاد 4. في الوقت الحاضر العلاج الوحيد غير داعمة لHIE هو انخفاض درجة حرارة الجسم بعد ميتة. وهكذا، والتقدم في إجراءات التشخيص واستراتيجيات العلاج ضرورية لتحسين إدارة HIE 8. وتستند تحسينات في التشخيص بعد الاختناق في الفترة المحيطة بالولادة وإدارة إصابات الدماغ حديثي الولادة على توسيع معرفة آليات المرض الكامنة فيالعلاجات الممكنة د. نماذج حيوانية من HIE مفيدة بشكل خاص عن الأحداث سريرية مختلفة قد تؤدي إلى HIE والإصابة في أي مركز ولادة واحد منخفض 5. الإعداد التجريبي الذي تأثير الاختلاف البيولوجي يمكن أن يكون الحد الأدنى، أمر ضروري عند اختبار جديد أدوات النذير والتشخيص واستراتيجيات العلاج. يجب على نموذج حيواني تقارب الحالة السريرية قدر الإمكان، وبالتالي المساهمة في فهم الآليات المرضية الكامنة وراء الإصابة التي يسببها وعملية ديناميكية تشارك في مرض وانها نتيجة 9. وشملت نماذج حيوانية من الأطفال حديثي الولادة HIE عدد من الأنواع، بما في ذلك القوارض، والضأن، والخنازير. في المقابل، خنزير صغير حديث الولادة لديها أعلى تشابه الوليد البشري فيما يتعلق حجم، ونظام القلب والأوعية الدموية 10 ونضج الدماغ في وقت التسليم 11،12. الرصد والقياس وتقييم النتائج في النموذج خنزير صغير يشبه ثاتي المستخدمة في الرعاية السريرية لالاطفال الذين يعانون من عجل. وفقا لذلك، هناك درجة عالية من الترجمة إلى رعاية الأطفال حديثي الولادة من هذا النموذج. وتستخدم نماذج الخنازير من نقص الأكسجة حول الولادة وعجل من قبل العديد من الجماعات وتختلف في عدد من مناطق 13. وفقا للغرض من هذه التجربة، ويجب إيلاء اهتمام دقيق لاختيار الأدوية وطريقة إحداث نقص الأكسجة نقص التروية، وطريقة التحكم في مدة وشدة الإهانة، الإنعاش بعد إهانة والرعاية، وتقييم النتائج. لتجنب التحيز وينبغي دائما على تصميم التجارب العشوائية يمكن استخدامها في الدراسات التدخلية. طريقة تطبيقها عند يحفز الإصابة بنقص التأكسج الدماغية هو المهم. نقص الأكسجة العالمي مما أدى إلى HIE غالبا ما يؤدي إلى فشل الأعضاء المتعددة التي تنطوي على الدماغ والقلب والرئتين والكلى والكبد. اعتمادا على نتائج تقييمها، ينبغي أن تستند نماذج من HIE على نقص الأكسجين ونقص التروية العالمي بدلا من الاعتماد على نقص التروية التنسيق، على سبيل المثال، من خلال ربط السيارةالشرايين otid 14. ورقة الأخيرة تطبيق مزيج من نقص الأكسجة (قوة المراقبة الدولية 2 12٪) والشريان السباتي ضغط الشريان مع الحفاظ على متوسط ​​ضغط الدم الشرياني&gt; 40 ملم زئبق 2. مجموعة أخرى يسببها نقص الأكسجة العالمي بنسبة 8٪ O 2 حتى القاعدة السلبية الزائدة&gt; 20 مليمول / لتر أو يعني ضغط الدم الشرياني (MABP) &lt;15 ملم زئبق، والتضحية الحيوانات في 4 ساعة 15. كما تم معاير نقص الأكسجة التي كتبها النتاج القلبي (30-40٪ من خط الأساس)، MABP (ل30-35 ملم زئبق) ودرجة الحموضة الشرياني (6،95-7،05) 16. وقد أظهرت نماذج العالمي نقص الأكسجة نقص التروية معاير التي كتبها aEEG قمع مماثلة لتلك الواردة في هذا التقرير، والاعتلال الدماغي الذي هو سريريا، electrophysiologically، وقابلة للمقارنة neuropathologically إلى حالة وجدت في الرضع المدى اختنق 17،18. درجة عجل بفعل أمر ضروري. ويجب على نموذج حيواني مفيد للHIE تسمح أيضا لاختبار ديا جديدإجراءات وخيارات العلاج معرفي. لتمكين هذا، ينبغي للنماذج لحث على عجل معتدل حيث هناك إمكانية العلاج في إصابات الدماغ الحادة مع إمكانية العلاج ضئيلة أو معدومة سيكون أقل أهمية عند تقييم علاجات جديدة. التسامح لنقص الأكسجين اختلافا كبيرا بين حيوانات التجارب. وقد أظهرت دراسات سابقة أن إصابات الدماغ أكثر اتساقا يمكن أن يتحقق وأن أكثر الحيوانات البقاء على قيد الحياة 17،19 من قبل إضفاء الطابع الشخصي على نقص الأكسجة الناجم فقا لاستجابة كل خنزير صغير في الدماغ تقييمها من قبل السعة متكاملة كهربية (aEEG) بدلا من استخدام مجموعة قوة المراقبة الدولية 2 القيمة في جميع أنحاء الحدث ميتة. مدة aEEG قمع يرتبط إلى درجة من إصابات الدماغ، مع بعض التغييرات النسيجية في &lt;20 دقيقة aEEG قمع والنوبات الدماغية الخطيرة زيادة في&gt; 45 دقيقة aEEG القمع. وقد أظهرت مراجعة الأخير من العلاجات اعصاب لHIE الحاجة إلى نماذج بقاء تمكين الاتفاقات البيئية المتعددة الأطراف النتيجة السلوكيةures في النماذج الحيوانية 20. هناك العديد من المزايا من HIE نموذج خنزير صغير المقدمة. لأنه يقوم على الأنواع التي يحتمل بشكل كبير لترجمة لعلم وظائف الأعضاء البشرية النتائج. نماذج نقص الأكسجة نقص التروية العالمية فشل الأعضاء المتعددة والمعايرة من نقص الأكسجة نقص التروية التي كتبها aEEG يؤدي الى درجة ثابتة من إصابات الدماغ مع بقاء النتائج السريرية ذات الصلة مثل أن المؤشرات الحيوية، التصوير بالرنين المغناطيسي والسلوك يمكن تقييمها في نقاط زمنية ذات الصلة. وقد نماذج خنزير صغير من الاختناق ما حول الولادة وعجل لم تسهم فقط إلى حد كبير في البصيرة الحالية إلى HIE الفيزيولوجيا المرضية، ولكن أيضا سبق بنجاح التجارب السريرية، مما يؤدي في نهاية المطاف إلى علاجات جديدة في البشر. لعبت دراسات نموذج الخنزير الصغير دورا رئيسيا في تأسيس انخفاض حرارة الجسم كعلاج للHIE 21، وتستخدم في البحوث إنعاش الوليد 22. وقد استخدمت مجموعات مختلفة نماذج خنزير صغير عند إجراء البحوث داخل الاختناق وعجل، وهووتشمل د الدراسات انخفاض حرارة الجسم 23، وتحفيز الخلايا الصباغية ألفا هرمون 24، وتوقف القلب 25، التيروزين النشاط هيدروكسيلاز 26، التعرض المتكرر ميتة 27، NMDA نشاط مستقبلات 14، والأشعة تحت الحمراء القريبة التحليل الطيفي 28. على عجل نموذج خنزير صغير الواردة في هذا التقرير يمثل تحديا تقنيا للعمل مع، وتعديلات طفيفة أثناء إجراء قد يؤدي إلى إما خفيفة جدا أو شديدة جدا إصابات الدماغ 29،2. وجدنا أن الكتابات الموجودة تفتقر إلى التفاصيل الكافية لإعادة إنتاج النماذج المنشورة سابقا. وبالتالي، نحن هنا لشرح كل خطوة من الإجراءات الفنية اللازمة لإنشاء خنزير صغير 72 ساعة نموذج البقاء على قيد الحياة في هذا التقرير، تمكين الباحثين من إقامة هذا النموذج المتقدم لدراسة عجل.

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a piglet model of neonatal hypoxic-ischemic encephalopathy

الاختناق أثناء الولادة، والذي يسبب بنقص التأكسج اعتلال الدماغ الإقفاري (عجل)، وحسابات لل0660000 شخص حول العالم سنويا، نحو ربع 2.9 مليون حالة وفاة حديثي الولادة في العالم. وقد ساهمت نماذج حيوانية من عجل في فهم الفيزيولوجيا المرضية في عجل، وسلطت الضوء على العملية الديناميكية التي تحدث في إصابات الدماغ بسبب الاختناق في الفترة المحيطة بالولادة. وهكذا، وقد اقترحت الدراسات الحيوان نافذة الوقت لاستراتيجيات العلاج بعد إهانة. وقد تم اختبار انخفاض حرارة الجسم كعلاج للعجل في نماذج pdiglet وأثبتت فعاليتها في التجارب السريرية في وقت لاحق. وقد طبقت الاختلافات في نموذج في دراسة طرق اعصاب مرافقة والدراسات خنزير صغير من الزينون والميلاتونين أدت إلى المرحلة السريرية I و II المحاكمات 1،2. وكذلك استخدم النموذج خنزير صغير عجل لresuscitation- والدورة الدموية دراسات حديثي الولادة وكذلك في التحقيقات من نقص الأكسجة الدماغية على المستوى الخلوي. ومع ذلك، فإنه هو تحديا تقنيانموذج والاختلافات في البروتوكول قد يؤدي إما خفيفة جدا أو شديدة جدا إصابات الدماغ. في هذه المقالة، ونحن لشرح الإجراءات الفنية اللازمة لإنشاء نموذج خنزير صغير مستقر من الأطفال حديثي الولادة عجل. أولا، يتم تخدير خنزير صغير حديث الولادة (&lt;24 ساعة من العمر، متوسط ​​وزن 1500 غ)، مدخل أنبوب، ورصد في إعداد مماثلة لتلك الموجودة في وحدة العناية المركزة لحديثي الولادة. هو فعل العالمي نقص الأكسجة نقص التروية عن طريق خفض نسبة الأكسجين الشهيق لتحقيق نقص الأكسجة العالمي، نقص التروية من خلال انخفاض ضغط الدم وEEG السعة أثر شقة متكاملة (aEEG) يدل على نقص الأكسجة الدماغية. يتم الترويج البقاء على قيد الحياة من خلال تعديل الأوكسجين وفقا لاستجابة aEEG وضغط الدم. وكميا إصابة الدماغ عن طريق التشريح والتصوير بالرنين المغناطيسي بعد 72 ساعة.

<html> يتم توثيق آثار نقص الأكسجين نقص التروية على الدماغ التي تحدث أثناء إهانة الناجم عن تسجيل التتبع aEEG. ويرد ممثل aEEG أثر في الشكل (6). <img alt="الشكل (6)" src="/files/ftp_upload/52454/52454fig6.jpg" /> الرقم 6. ممثل aEEG أثر.</...

Watch this Scientific Journal Video about A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy at JoVE.com

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

A الخنزير الصغير نموذج من الأطفال حديثي الولادة نقص الأوكسجين، اعتلال الدماغ الإقفاري

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the ...

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19.5K Views.  Institute of Clinical Medicine, Aarhus University Hospital. Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy. 

Video: A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

A Swine Model of Neonatal Asphyxia

Annually more than 1 million neonates die worldwide as related to asphyxia. Asphyxiated neonates commonly have multi-organ failure including hypotension, perfusion deficit, hypoxic-ischemic encephalopathy, pulmonary hypertension, vasculopathic enterocolitis, renal failure and thrombo-embolic complications. Animal models are developed to help us understand the patho-physiology and pharmacology of neonatal asphyxia. In comparison to rodents and newborn lambs, the newborn piglet has been proven to be a valuable model. The newborn piglet has several advantages including similar development as that of 36-38 weeks human fetus with comparable body systems, large body size (&tilde;1.5-2 kg at birth) that allows the instrumentation and monitoring of the animal and controls the confounding variables of hypoxia and hemodynamic derangements.
We here describe an experimental protocol to simulate neonatal asphyxia and allow us to examine the systemic and regional hemodynamic changes during the asphyxiating and reoxygenation process as well as the respective effects of interventions. Further, the model has the advantage of studying multi-organ failure or dysfunction simultaneously and the interaction with various body systems. The experimental model is a non-survival procedure that involves the surgical instrumentation of newborn piglets (1-3 day-old and 1.5-2.5 kg weight, mixed breed) to allow the establishment of mechanical ventilation, vascular (arterial and central venous) access and the placement of catheters and flow probes (Transonic Inc.) for the continuously monitoring of intra-vascular pressure and blood flow across different arteries including main pulmonary, common carotid, superior mesenteric and left renal arteries. Using these surgically instrumented piglets, after stabilization for 30-60 minutes as defined by Z&lt;10% variation in hemodynamic parameters and normal blood gases, we commence an experimental protocol of severe hypoxemia which is induced via normocapnic alveolar hypoxia. The piglet is ventilated with 10-15% oxygen by increasing the inhaled concentration of nitrogen gas for 2h, aiming for arterial oxygen saturations of 30-40%. This degree of hypoxemia will produce clinical asphyxia with severe metabolic acidosis, systemic hypotension and cardiogenic shock with hypoperfusion to vital organs. The hypoxia is followed by reoxygenation with 100% oxygen for 0.5h and then 21% oxygen for 3.5h. Pharmacologic interventions can be introduced in due course and their effects investigated in a blinded, block-randomized fashion.

Large animal models have good translational values in the examination of physiology and pharmacology of neonatal asphyxia. Using newborn piglets, we develop an experimental protocol to simulate neonatal asphyxia which has advantages of studying the systemic and regional hemodynamics, oxygen transport with biochemical and pathologic pathways and correlations.

نموذج الخنازير من الاختناق الوليدي

Annually more than 1 million neonates die worldwide as related to asphyxia. Asphyxiated neonates commonly have multi-organ failure including hypotension, ...

Assessment of Perigenital Sensitivity and Prostatic Mast Cell Activation in a Mouse Model of Neonatal Maternal Separation

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has a lifetime prevalence of 14% and is the most common urological diagnosis for men under the age of 50, yet it is the least understood and studied chronic pelvic pain disorder. A significant subset of patients with chronic pelvic pain report having experienced early life stress or abuse, which can markedly affect the functioning and regulation of the hypothalamic-pituitary-adrenal (HPA) axis. Mast cell activation, which has been shown to be increased in both urine and expressed prostatic secretions of CP/CPPS patients, is partially regulated by downstream activation of the HPA axis. Neonatal maternal separation (NMS) has been used for over two decades to study the outcomes of early life stress in rodent models, including changes in the HPA axis and visceral sensitivity. Here we provide a detailed protocol for using NMS as a preclinical model of CP/CPPS in male C57BL/6 mice. We describe the methodology for performing NMS, assessing perigenital mechanical allodynia, and histological evidence of mast cell activation. We also provide evidence that early psychological stress can have long-lasting effects on the male urogenital system in mice.

We are measuring perigenital mechanical sensitivity and mast cell activation in the prostate of male C57BL/6 mice that underwent an early life stress paradigm &#8211; neonatal maternal separation, in order to induce a preclinical model of chronic prostatitis/chronic pelvic pain syndrome.

تقييم محيط بالغدد التناسلية حساسية والبروستات ماست تنشيط خلية في نموذج الفأر من فصل الأمهات حديثي الولادة

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has a lifetime prevalence of 14% and is the most common urological diagnosis for men under the ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

وحديثي الولادة ماوس الحبل الشوكي ضغط إصابة نموذج

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

A Battery of Motor Tests in a Neonatal Mouse Model of Cerebral Palsy

As the sheer number of transgenic mice strains grow and rodent models of pediatric disease increase, there is an expanding need for a comprehensive, standardized battery of neonatal mouse motor tests. These tests can validate injury or disease models, determine treatment efficacy and/or assess motor behaviors in new transgenic strains. This paper presents a series of neonatal motor tests to evaluate general motor function, including ambulation, hindlimb foot angle, surface righting, negative geotaxis, front- and hindlimb suspension, grasping reflex, four limb grip strength and cliff aversion. Mice between the ages of post-natal day 2 to 14 can be used. In addition, these tests can be used for a wide range of neurological and neuromuscular pathologies, including cerebral palsy, hypoxic-ischemic encephalopathy, traumatic brain injury, spinal cord injury, neurodegenerative diseases, and neuromuscular disorders. These tests can also be used to determine the effects of pharmacological agents, as well as other types of therapeutic interventions. In this paper, motor deficits were evaluated in a novel neonatal mouse model of cerebral palsy that combines hypoxia, ischemia and inflammation. Forty-eight hours after injury, five tests out of the nine showed significant motor deficits: ambulation, hindlimb angle, hindlimb suspension, four limb grip strength, and grasping reflex. These tests revealed weakness in the hindlimbs, as well as fine motor skills such as grasping, which are similar to the motor deficits seen in human cerebral palsy patients.

Presented is a concise battery of mouse neonatal motor tests. Using these tests, neonatal motor deficits can be demonstrated in a variety of neonatal motor disorders. By having a standardized set of tests, results from different studies can be compared, allowing for better and accurate reporting between groups.

مجموعة من الاختبارات للسيارات في ماوس نموذج حديثي الولادة من الشلل الدماغي

As the sheer number of transgenic mice strains grow and rodent models of pediatric disease increase, there is an expanding need for a comprehensive, ...

Myocardial Infarction in Neonatal Mice, A Model of Cardiac Regeneration

Myocardial infarction induced by coronary artery ligation has been used in many animal models as a tool to study the mechanisms of cardiac repair and regeneration, and to define new targets for therapeutics. For decades, models of complete heart regeneration existed in amphibians and fish, but a mammalian counterpart was not available. The recent discovery of a postnatal window during which mice possess regenerative capabilities has led to the establishment of a mammalian model of cardiac regeneration. A surgical model of mammalian cardiac regeneration in the neonatal mouse is presented herein. Briefly, postnatal day 1 (P1) mice are anesthetized by isoflurane and placed on an ice pad to induce hypothermia. After the chest is opened, and the left anterior descending coronary artery (LAD) is visualized, a suture is placed around the LAD to inflict myocardial ischemia in the left ventricle. The surgical procedure takes 10-15 min. Visualizing the coronary artery is crucial for accurate suture placement and reproducibility. Myocardial infarction and cardiac dysfunction are confirmed by triphenyl-tetrazolium chloride (TTC) staining and echocardiography, respectively. Complete regeneration 21 days post myocardial infarction is verified by histology. This protocol can be used to as a tool to elucidate mechanisms of mammalian cardiac regeneration after myocardial infarction.

This protocol describes a highly reproducible model of cardiac regeneration by surgical induction of myocardial infarction in the left ventricle of postnatal day 1 mice. The method involves induction of hypothermic anesthesia and ligation of the left anterior descending coronary artery.

احتشاء عضلة القلب في الفئران حديثي الولادة، نموذج للتجديد القلب

Myocardial infarction induced by coronary artery ligation has been used in many animal models as a tool to study the mechanisms of cardiac repair and ...

Use of a Piglet Model for the Study of Anesthetic-induced Developmental Neurotoxicity (AIDN): A Translational Neuroscience Approach

Anesthesia cannot be avoided in many cases when surgery is required, particularly in children. Recent investigations in animals have raised concerns that anesthesia exposure may lead to neuronal apoptosis, known as anesthesia-induced developmental neurotoxicity (AIDN). Furthermore, some clinical studies in children have suggested that anesthesia exposure may lead to neurodevelopmental deficits later in life. Nonetheless, an ideal animal model for preclinical study has yet to be developed. The neonatal piglet represents a valuable model for preclinical study, as they share a striking number of developmental similarities with humans.
The anatomy and physiology of piglets allow for implementation of rigorous human perioperative conditions in both survival and non-survival procedures. Femoral artery catheterization allows for close monitoring, thus enabling prompt correction of any deviation of the piglet's vital signs and chemistries. In addition, there are multiple developmental similarities between piglets and human neonates. The techniques required to use piglets for experimentation will require experience to master. A pediatric anesthesiologist is a critical member of the investigative team. We describe, in a general sense, the appropriate use of a piglet model for neurodevelopmental study.

Use of a Piglet Model for the Study of Anesthetic-induced Developmental Neurotoxicity AIDN: A Translational Neuroscience Approach

Anesthesia-induced developmental neurotoxicity (AIDN) research has focused on rodents, which are not broadly applicable to humans. Non-human primate models are more relevant, but are cost-prohibitive and difficult to use for experimentation. The piglet, in contrast, is a clinically relevant, practical animal model ideal for the study of anesthetic neurotoxicity.

استخدام نموذج خنزير صغير لدراسة التسمم العصبي التنموي الناجم عن التخدير (إيدن): نهج علم الأعصاب الانتقالي

Anesthesia cannot be avoided in many cases when surgery is required, particularly in children.  Recent investigations in animals have raised concerns that ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

متساوي القياس كونتراكتيليتي القياس الشريان المساريقي العلوي الماوس باستخدام أسلاك ميوجرافي

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

A Neonatal BALB/c Mouse Model of Necrotizing Enterocolitis

Necrotizing enterocolitis (NEC) is the most severe gastrointestinal (GI) disease that often occurs in premature infants, especially very low birth weight infants, with high mortality and unclear pathogenesis. The cause of NEC may be related to inflammatory immune regulatory system abnormalities. An NEC animal model is an indispensable tool for NEC disease immune research. NEC animal models usually use C57BL/6J neonatal mice; BALB/c neonatal mice are rarely used. Related studies have shown that when mice are infected, Th2 cell differentiation is predominant in BALB/c mice compared to C57BL/6J mice. Studies have suggested that the occurrence and development of NEC are associated with an increase in T helper type 2 (Th2) cells and are generally accompanied by infection. Therefore, this study used neonatal BALB/c mice to induce an NEC model with similar clinical characteristics and intestinal pathological changes as those observed in children with NEC. Further study is warranted to determine whether this animal model could be used to study Th2 cell responses in NEC.

Necrotizing enterocolitis (NEC) is the most severe gastrointestinal (GI) disease that often occurs in premature infants, especially very low birth weight infants, with high mortality and unclear pathogenesis. The cause of NEC may be related to inflammatory immune regulatory system abnormalities. An NEC animal model is an indispensable tool for NEC disease immune research. NEC animal models usually use C57BL/6J neonatal mice; BALB/c neonatal mice are rarely used. Related studies have shown that when mice are infected, Th2 cell differentiation is predominant in BALB/c mice compared to C57BL/6J mice. Studies have suggested that the occurrence and development of NEC are associated with an increase in T helper type 2 (Th2) cells and are generally accompanied by infection. Therefore, this study used neonatal BALB/c mice to induce an NEC model with similar clinical characteristics and intestinal pathological changes as those observed in children with NEC. Further study is warranted to determine whether this animal model could be used to study Th2 cell responses in NEC.

نموذج فأر BALB/c الوليدي لالتهاب الأمعاء والقولون الناخر

Necrotizing enterocolitis (NEC) is the most severe gastrointestinal (GI) disease that often occurs in premature infants, especially very low birth weight ...

Effect of Hyaluronic Acid 35 kDa on an In Vitro Model of Preterm Small Intestinal Injury and Healing Using Enteroid-Derived Monolayers

In vitro scratch wound assays are commonly used to investigate the mechanisms and characteristics of epithelial healing in a variety of tissue types. Here, we describe a protocol to generate a two-dimensional (2D) monolayer from three-dimensional (3D) non-human primate enteroids derived from intestinal crypts of the terminal ileum. These enteroid-derived monolayers were then utilized in an in vitro scratch wound assay to test the ability of hyaluronan 35 kDa (HA35), a human milk HA mimic, to promote cell migration and proliferation along the epithelial wound edge. After the monolayers were grown to confluency, they were manually scratched and treated with HA35 (50 &#181;g/mL, 100 &#181;g/mL, 200 &#181;g/mL) or control (PBS). Cell migration and proliferation into the gap were imaged using a transmitted-light microscope equipped for live-cell imaging. Wound closure was quantified as percent wound healing using the Wound Healing Size Plugin in ImageJ.&#160;The scratch area and rate of cell migration and the percentage of wound closure were measured over 24 h. HA35 in vitro accelerates wound healing in small intestinal enteroid monolayers, likely through a combination of cell proliferation at the wound edge and migration to the wound area. These methods can potentially be used as a model to explore intestinal regeneration in the preterm human small intestine.

Effect of Hyaluronic Acid 35 kDa on an In Vitro Model of Preterm Small Intestinal Injury and Healing Using Enteroid-Derived Monolayers

This protocol describes a method to establish and perform a scratch wound assay on two-dimensional (2D) monolayers derived from three-dimensional (3D) enteroids isolated from non-human primate ileum.

تأثير حمض الهيالورونيك 35 كيلو دال أمبير على نموذج في المختبر للإصابات المعوية الصغيرة المبكرة والشفاء باستخدام الطبقات الأحادية المشتقة من الأمعاء

In vitro scratch wound assays are commonly used to investigate the mechanisms and characteristics of epithelial healing in a variety of tissue types. ...

In Vitro Apical-Out Enteroid Model of Necrotizing Enterocolitis

Necrotizing enterocolitis (NEC) is a devastating disease affecting preterm infants, characterized by intestinal inflammation and necrosis. Enteroids have recently emerged as a promising system to model gastrointestinal pathologies. However, currently utilized methods for enteroid manipulation either lack access to the apical surface of the epithelium (three-dimensional [3D]) or are time-consuming and resource-intensive (two-dimensional [2D] monolayers). These methods often require additional steps, such as microinjection, for the model to become physiologically translatable. Here, we describe a physiologically relevant and inexpensive protocol for studying NEC in vitro by reversing enteroid polarity, resulting in the apical surface facing outward (apical-out). An immunofluorescent staining protocol to examine enteroid barrier integrity and junctional protein expression following exposure to tumor necrosis factor-alpha (TNF-&#945;) or lipopolysaccharide (LPS) under normoxic or hypoxic conditions is also provided. The viability of 3D apical-out enteroids exposed to normoxic or hypoxic LPS or TNF-&#945; for 24 h is also evaluated. Enteroids exposed to either LPS or TNF-&#945;, in combination with hypoxia, exhibited disruption of epithelial architecture, a loss of adherens junction protein expression, and a reduction in cell viability. This protocol describes a new apical-out NEC-in-a-dish model which presents a physiologically relevant and cost-effective platform to identify potential epithelial targets for NEC therapies and study the preterm intestinal response to therapeutics.

In Vitro Apical-Out Enteroid Model of Necrotizing Enterocolitis

This protocol describes an apical-out necrotizing enterocolitis (NEC)-in-a-dish model utilizing small intestinal enteroids with reversed polarity, allowing access to the apical surface. We provide an immunofluorescent staining protocol to detect NEC-related epithelial disruption and a method to determine the viability of apical-out enteroids subjected to the NEC-in-a-dish protocol.

في المختبر نموذج معوي قمي من التهاب الأمعاء والقولون الناخر

Necrotizing enterocolitis (NEC) is a devastating disease affecting preterm infants, characterized by intestinal inflammation and necrosis. Enteroids have ...