ويتم تمويل هذا العمل من قبل المعاهد الوطنية للصحة NR011209 R01 - 03

<ol>
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I., Sustrova, M., Garaiova, I., Muchova, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uric+acid+and+allantoin+levels+in+Down+syndrome:+antioxidant+and+oxidative+stress+mechanisms?.">Uric acid and allantoin levels in Down syndrome: antioxidant and oxidative stress mechanisms?.</a> Clin Chim Acta. 341, 139-146 (2004).</li><li>Siciarz, A., Weinberger, B., Witz, G., Hiatt, M., Hegyi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Urinary+thiobarbituric+acid-reacting+substances+as+potential+biomarkers+of+intrauterine+hypoxia.">Urinary thiobarbituric acid-reacting substances as potential biomarkers of intrauterine hypoxia.</a> Arch Pediatr Adolesc Med. 155, 718-722 (2001).</li><li>Buonocore, G., Perrone, S., Longini, M., Terzuoli, L., Bracci, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Total+hydroperoxide+and+advanced+oxidation+protein+products+in+preterm+hypoxic+babies.">Total hydroperoxide and advanced oxidation protein products in preterm hypoxic babies.</a> Pediatr Res. 47, 221-224 (2000).</li><li>Berne, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+nucleotides+in+hypoxia:+possible+role+in+regulation+of+coronary+blood+flow.">Cardiac nucleotides in hypoxia: possible role in regulation of coronary blood flow.</a> Am J Physiol. 204, 317-322 (1963).</li><li>Harkness, R. A., Lund, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebrospinal+fluid+concentrations+of+hypoxanthine,+xanthine,+uridine+and+inosine:+high+concentrations+of+the+ATP+metabolite,+hypoxanthine,+after+hypoxia.">Cerebrospinal fluid concentrations of hypoxanthine, xanthine, uridine and inosine: high concentrations of the ATP metabolite, hypoxanthine, after hypoxia.</a> J Clin Pathol. 36, 1-8 (1983).</li><li>Plank, M. S., Boskovic, D. S., Sowers, L. C., Angeles, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biochemical+markers+of+neonatal+hypoxia.">Biochemical markers of neonatal hypoxia.</a> Pediatric Health. 2, 485-501 (2008).</li><li>Cighetti, G., Allevi, P., Anastasia, L., Bortone, L., Paroni, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+methyl+malondialdehyde+as+an+internal+standard+for+malondialdehyde+detection:+validation+by+isotope-dilution+gas+chromatography-mass+spectrometry.">Use of methyl malondialdehyde as an internal standard for malondialdehyde detection: validation by isotope-dilution gas chromatography-mass spectrometry.</a> Clin Chem. 48, 2266-2269 (2002).</li><li>Paroni, R., Fermo, I., Cighetti, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Validation+of+methyl+malondialdehyde+as+internal+standard+for+malondialdehyde+detection+by+capillary+electrophoresis.">Validation of methyl malondialdehyde as internal standard for malondialdehyde detection by capillary electrophoresis.</a> Anal Biochem. 307, 92-98 (2002).</li><li>Cighetti, G., Debiasi, S., Ciuffreda, P., Allevi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beta-ethoxyacrolein+contamination+increases+malondialdehyde+inhibition+of+milk+xanthine+oxidase+activity.">Beta-ethoxyacrolein contamination increases malondialdehyde inhibition of milk xanthine oxidase activity.</a> Free Radic Biol Med. 25, 818-825 (1998).</li><li>Cighetti, G., Debiasi, S., Paroni, R., Allevi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Free+and+total+malondialdehyde+assessment+in+biological+matrices+by+gas+chromatography-mass+spectrometry:+what+is+needed+for+an+accurate+detection.">Free and total malondialdehyde assessment in biological matrices by gas chromatography-mass spectrometry: what is needed for an accurate detection.</a> Anal Biochem. 266, 222-229 (1999).</li><li>Yamamoto, T., Moriwaki, Y., Takahashi, S., Tsutsumi, Z., Yamakita, J., Nasako, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+human+plasma+xanthine+oxidase+activity+by+high-performance+liquid+chromatography.">Determination of human plasma xanthine oxidase activity by high-performance liquid chromatography.</a> J Chromatogr B Biomed Appl. 681, 395-400 (1996).</li><li>Fasman, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of Biochemistry and Molecular Biology. , (1988).</li><li>Chen, X. B., Calder, A. G., Prasitkusol, P., Kyle, D. J., Jayasuriya, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+15N+isotopic+enrichment+and+concentrations+of+allantoin+and+uric+acid+in+urine+by+gas+chromatography/mass+spectrometry.">Determination of 15N isotopic enrichment and concentrations of allantoin and uric acid in urine by gas chromatography/mass spectrometry.</a> J Mass Spectrom. 33, 130-137 (1998).</li></ol>

الإعلان عن أي تضارب في المصالح.

وصفت وسائل تسمح هنا تقييم نقص التروية نقص الأكسجة الولدان. هذا البروتوكول يجمع بين قياسات علامات الحرمان (ATP) والطاقة ، والاكسدة ، والضرر التأكسدي ، ونشاط انزيم للحصول على صورة شاملة للوجود البيوكيميائية أو حتى درجة من نقص تروية ميتة. على الرغم من جدوى هذا الأسلوب ، و?...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><th> اسم كاشف </th><th> شركة </th><th> فهرس العدد </th><th> تعليقات (اختياري) </th></tr><tr><td> 6ml K3E أنبوب EDTA K3 </td><td> فيشر العلمية </td><td> 2204061 </td><td></td></tr><tr><td> 5702R الطرد المركزي </td><td> فيشر العلمية </td><td> 05413319 </td><td> مع 13 و محول 16MM </td></tr><tr><td> 1.5ml microcentrifuge أنبوب </td><td> الولايات المتحدة الأمريكية والعلم </td><td> 1615-5599 </td><td></td></tr><tr><td> 2 - Aminopurine </td><td> سيغما الدريخ </td><td> A3509 </td><td></td></tr><tr><td> فاريان كاري معمل 100 </td><td> Agilant تكنولوجيز </td><td> 0010071500 </td><td></td></tr><tr><td> موهوب SpeedVac </td><td> الحرارية العلمية </td><td> SC210A - 115 </td><td></td></tr><tr><td> جهاز الطرد المركزي فلتر ميكرون </td><td> فيشر العلمية </td><td> UFC501596 </td><td></td></tr><tr><td> Supelcosil LC - 18 - S العمود </td><td> سيغما الدريخ </td><td> 58931 </td><td></td></tr><tr><td> Supelcosil LC - 18 - S Supelguard وحامل خرطوشة </td><td> سيغما الدريخ </td><td> 59629 </td><td></td></tr><tr><td> HPLC </td><td> مياه </td><td></td><td></td></tr><tr><td> هذه النماذج فيال </td><td> فيشر العلمية </td><td> 03376607 </td><td></td></tr><tr><td> DL - آلانتوين C - 5 -13 (1) ؛ -15 N </td><td> CDN نظائر </td><td> M - 2307 </td><td> # الكثير L340P9 </td></tr><tr><td> MTBSTFA </td><td> الحرارية العلمية </td><td> 48920 </td><td></td></tr><tr><td> بيريدين </td><td> سيغما الدريخ </td><td> 270970 </td><td></td></tr><tr><td> 5973E GC / MSD </td><td> اجيلنت تكنولوجيز </td><td> G7021A </td><td> جزء # ل5975E GC / MS </td></tr><tr><td> 3 Ethoxymethacrolein </td><td> سيغما الدريخ </td><td> 232548 </td><td></td></tr><tr><td> هيدروكسيد الصوديوم </td><td> سيغما الدريخ </td><td> S5881 </td><td></td></tr><tr><td> ثنائي كلورو ميثان </td><td> سيغما الدريخ </td><td> 270997 </td><td></td></tr><tr><td> البنزين </td><td> سيغما الدريخ </td><td> 401765 </td><td></td></tr><tr><td> Diisopropyl الأثير </td><td> سيغما الدريخ </td><td> 38270 </td><td></td></tr><tr><td> BHT </td><td> سيغما الدريخ </td><td> B1378 </td><td></td></tr><tr><td> الإيثانول </td><td> سيغما الدريخ </td><td> 459844 </td><td></td></tr><tr><td> الفينيل هيدرازين </td><td> سيغما الدريخ </td><td> P26252 </td><td></td></tr></tbody></table>

biochemical measurement of neonatal hypoxia

وتتميز الوليدية الإسكيمية نقص الأكسجين التي نضح الدم غير كافية من الأنسجة أو عدم نظامية من الأوكسجين. ويعتقد أن هذا الشرط لسبب / تفاقم الاضطرابات الولدان موثقة جيدا بما في ذلك إعاقة للجهاز العصبي 1-3. انخفض انتاج الأدينوساين ثلاثي يحدث نتيجة لنقص في الفسفرة المؤكسدة. للتعويض عن هذه الجزيئات التي تحتوي على طاقة عالية حرمان الدولة سندات الفوسفات الطاقة هي 2 المتدهورة. هذا يؤدي إلى زيادة مستويات الأدينوساين الذي تدهورت لاحقا إينوزين ، هيبوزانتين ، الزانثين ، وأخيرا إلى حمض اليوريك. يتم تنفيذ المرحلتين الأخيرتين في هذه العملية من قبل تدهور oxidoreductase الزانثين. هذا الانزيم موجود في شكل هيدروجين الزانثين في ظل ظروف normoxic ولكن يتم تحويلها إلى أوكسيديز الزانثين (XO) تحت ظروف نقص الأكسجين ضخه - 4 ، 5. خلافا نازعة الزانثين ، XO يولد بيروكسيد الهيدروجين كمنتج ثانوي للتدهور البيورين 4 ، 6. هذا بيروكسيد الهيدروجين في الجمع مع غيرها من أنواع الاكسجين التفاعلية (ريوس) التي تنتج أثناء نقص الأكسجين ، يتأكسد حمض اليوريك لتشكيل آلانتوين ويتفاعل مع الأغشية الدهنية لتوليد malondialdehyde (MDA) 7-9 ، ومعظم الثدييات ، البشر المعفاة ، تمتلك يوريكاز الانزيم الذي يحول إلى حمض اليوريك آلانتوين. في البشر ، ومع ذلك ، لا يمكن إلا أن يكون شكلتها آلانتوين ريوس بوساطة أكسدة حمض اليوريك. وبسبب هذا ، يعتبر آلانتوين أن يكون علامة من الاكسدة في البشر ، ولكن ليس في الثدييات التي لديها يوريكاز. نحن تصف أساليب توظيف ارتفاع ضغط اللوني السائل (HPLC) والغاز اللوني الطيف الكتلي (نماذج الدوران) لقياس العلامات الحيوية من نقص التروية نقص الأكسجين الوليدي ، ويستخدم الدم البشرية لمعظم الاختبارات. ويمكن أيضا أن تستخدم دم الحيوان مع الاعتراف احتمال يوريكاز المولدة آلانتوين. وترتبط الأيضات البيورين إلى نقص الأكسجين في وقت مبكر عام 1963 وموثوقية هيبوزانتين ، الزانثين ، وجرى التحقق من حمض اليوريك كمؤشرات البيوكيميائية حديثي الولادة من نقص الأكسجة المحققون عدة 10-13. طريقة HPLC المستخدمة في القياس الكمي للمركبات البيورين سريعة وموثوق بها ، وقابلة للتكرار. تم تعديل طريقة GC / MS المستخدمة في القياس الكمي للآلانتوين ، علامة جديدة نسبيا من الاكسدة ، وآخرون من غروبر 7. هذا الأسلوب يتجنب بعض التحف ويتطلب كميات منخفضة من العينة. ووصف الأساليب المستخدمة لتوليف MMDA في أماكن أخرى 14 و 15. تم تكييفها GC / MS الكمي القائم على نجمة داود الحمراء من Paroni وآخرون. وCighetti آخرون 16 و 17. تم قياس النشاط أوكسيديز الزانثين بواسطة HPLC عن طريق قياس تحويل تيرين isoxanthopterin إلى 18. أثبت هذا النهج لتكون حساسة بما فيه الكفاية وقابلة للتكرار.

Watch this Scientific Journal Video about Biochemical Measurement of Neonatal Hypoxia at JoVE.com

Biochemical Measurement of Neonatal Hypoxia

تم وصف طريقة لقياس علامات الحيوية في الأطفال حديثي الولادة نقص الأكسجين ، نقص التروية. هذا النهج يستخدم ارتفاع ضغط اللوني السائل (HPLC) واللوني للغاز طيف الكتلة (GC / MS).

البيوكيميائية قياس حديثي الولادة نقص الأكسجة

Neonatal hypoxia ischemia is characterized by inadequate blood perfusion of a tissue or a systemic lack of oxygen. This condition is thought to ...

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14.0K Views.  Loma Linda University. تم وصف طريقة لقياس علامات الحيوية في الأطفال حديثي الولادة نقص الأكسجين ، نقص التروية. هذا النهج يستخدم ارتفاع ضغط اللوني السائل (HPLC) واللوني للغاز طيف الكتلة (GC / MS).

Video: Biochemical Measurement of Neonatal Hypoxia

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, ...

Mouse Model of Intraluminal MCAO: Cerebral Infarct Evaluation by Cresyl Violet Staining

Stroke is the third cause of mortality and the leading cause of disability in the World. Ischemic stroke accounts for approximately 80% of all strokes. However, the thrombolytic tissue plasminogen activator (tPA) is the only treatment of acute ischemic stroke that exists. This led researchers to develop several ischemic stroke models in a variety of species. Two major types of rodent models have been developed: models of global cerebral ischemia or focal cerebral ischemia. To mimic ischemic stroke in patients, in whom approximately 80% thrombotic or embolic strokes occur in the territory of the middle cerebral artery (MCA), the intraluminal middle cerebral artery occlusion (MCAO) model is quite relevant for stroke studies. This model was first developed in rats by Koizumi et al. in 1986 1. Because of the ease of genetic manipulation in mice, these models have also been developed in this species 2-3.
	Herein, we present the transient MCA occlusion procedure in C57/Bl6 mice. Previous studies have reported that physical properties of the occluder such as tip diameter, length, shape, and flexibility are critical for the reproducibility of the infarct volume 4. Herein, a commercial silicon coated monofilaments (Doccol Corporation) have been used. Another great advantage is that this monofilament reduces the risk to induce subarachnoid hemorrhages. Using the Zeiss stereo-microscope Stemi 2000, the silicon coated monofilament was introduced into the internal carotid artery (ICA) via a cut in the external carotid artery (ECA) until the monofilament occludes the base of the MCA. Blood flow was restored 1 hour later by removal of the monofilament to mimic the restoration of blood flow after lysis of a thromboembolic clot in humans. The extent of cerebral infarct may be evaluated first by a neurologic score and by the measurement of the infarct volume. Ischemic mice were thus analyzed for their neurologic score at different post-reperfusion times. To evaluate the infarct volume, staining with 2,3,5-triphenyltetrazolium chloride (TTC) was usually performed. Herein, we used cresyl violet staining since it offers the opportunity to test many critical markers by immunohistochemistry. In this video, we report the MCAO procedure; neurological scores and the evaluation of the infarct volume by cresyl violet staining.

The intraluminal middle cerebral occlusion model in mice is herein presented. The extent of cerebral infarct is evaluated by a neurologic score and cresyl violet staining, an alternative staining to TTC, offering the great advantage to test in parallel many interest markers.

نموذج الفأر من MCAO داخل اللمعة: الدماغي التقييم عائق من قبل تلوين البنفسج الكريزيل

Stroke is the third  cause of mortality and the leading cause of disability in the World. Ischemic  stroke accounts for approximately 80% of all strokes. ...

Non-invasive Optical Measurement of Cerebral Metabolism and Hemodynamics in Infants

Perinatal brain injury remains a significant cause of infant mortality and morbidity, but there is not yet an effective bedside tool that can accurately screen for brain injury, monitor injury evolution, or assess response to therapy. The energy used by neurons is derived largely from tissue oxidative metabolism, and neural hyperactivity and cell death are reflected by corresponding changes in cerebral oxygen metabolism (CMRO2). Thus, measures of CMRO2 are reflective of neuronal viability and provide critical diagnostic information, making CMRO2 an ideal target for bedside measurement of brain health.
Brain-imaging techniques such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT) yield measures of cerebral glucose and oxygen metabolism, but these techniques require the administration of radionucleotides, so they are used in only the most acute cases.
Continuous-wave near-infrared spectroscopy (CWNIRS) provides non-invasive and non-ionizing radiation measures of hemoglobin oxygen saturation (SO2) as a surrogate for cerebral oxygen consumption. However, SO2 is less than ideal as a surrogate for cerebral oxygen metabolism as it is influenced by both oxygen delivery and consumption. Furthermore, measurements of SO2 are not sensitive enough to detect brain injury hours after the insult 1,2, because oxygen consumption and delivery reach equilibrium after acute transients 3. We investigated the possibility of using more sophisticated NIRS optical methods to quantify cerebral oxygen metabolism at the bedside in healthy and brain-injured newborns. More specifically, we combined the frequency-domain NIRS (FDNIRS) measure of SO2 with the diffuse correlation spectroscopy (DCS) measure of blood flow index (CBFi) to yield an index of CMRO2 (CMRO2i) 4,5.
With the combined FDNIRS/DCS system we are able to quantify cerebral metabolism and hemodynamics. This represents an improvement over CWNIRS for detecting brain health, brain development, and response to therapy in neonates. Moreover, this method adheres to all neonatal intensive care unit (NICU) policies on infection control and institutional policies on laser safety. Future work will seek to integrate the two instruments to reduce acquisition time at the bedside and to implement real-time feedback on data quality to reduce the rate of data rejection.

We combined frequency-domain near-infrared spectroscopy measures of cerebral hemoglobin oxygenation with diffuse correlation spectroscopy measures of cerebral blood flow index to estimate an index of oxygen metabolism. We tested the utility of this measure as a bedside screening tool to evaluate the health and development of the newborn brain.

غير الغازية قياس بصري من الأيض الدماغي وديناميكا الدم في الأطفال الرضع

Perinatal brain injury remains a significant cause of infant mortality  and morbidity, but there is not yet an effective bedside tool that can  accurately ...

Collecting And Measuring Wound Exudate Biochemical Mediators In Surgical Wounds

We describe a methodology by which we are able to collect and measure biochemical inflammatory and nociceptive mediators at the surgical wound site. Collecting site-specific biochemical markers is important to understand the relationship between levels in serum and surgical wound, determine any associations between mediator release, pain, analgesic use and other outcomes of interest, and evaluate the effect of systemic and peripheral drug administration on surgical wound biochemistry. This methodology has been applied to healthy women undergoing elective cesarean delivery with spinal anesthesia. We have measured wound exudate and serum mediators at the same time intervals as patient's pain scores and analgesics consumption for up to 48 hours post-cesarean delivery. Using this methodology we have been able to detect various biochemical mediators including nerve growth factor (NGF), prostaglandin E2 (PG-E2) substance P, IL-1&#x03B2;, IL-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, TNF&alpha;, INF&#x03B3;, G-CSF, GM-CSF, MCP-1 and MIP-1&#x03B2;. Studies applying this human surgical wound bioassay have found no correlations between wound and serum cytokine concentrations or their time-release profile (J Pain. 2008; 9(7):650-7).1 We also documented the utility of the technique to identify drug-mediated changes in wound cytokine content (Anesth Analg 2010; 111:1452-9).2

This article provides a detailed and visual description of a methodology for collecting and measuring biochemical inflammatory and nociceptive mediators at the surgical wound site following cesarean delivery. This human bioassay has been used to determine correlations between wound and serum cytokine concentrations and drug-mediated changes in wound cytokines, chemokines and neuropetides.

جمع وقياس وسطاء الافرازات الكيميائية الحيوية في الجرح الجروح الجراحية

We describe a methodology by which we are able to collect and measure biochemical inflammatory and nociceptive mediators at the surgical wound site. ...

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease and injury regional estimates, World Health Organization, 2004). Surgeries that often compromise the quality of life are required to correct heart defects, reminding us of the importance of finding the causes of CHD. Mutant mouse models and live imaging technology have become essential tools to study the etiology of this disease. Although advanced methods allow live imaging of abnormal hearts in embryos, the physiological and hemodynamic states of the latter are often compromised due to surgical and/or lengthy procedures. Noninvasive ultrasound imaging, however, can be used without surgically exposing the embryos, thereby maintaining their physiology. Herein, we use simple M-mode ultrasound to assess heart rates of embryos at E18.5 in utero. The detection of abnormal heart rates is indeed a good indicator of dysfunction of the heart and thus constitutes a first step in the identification of developmental defects that may lead to heart failure.

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Ultra-high frequency ultrasound is a powerful live imaging tool to examine cardiac abnormalities in small animals. Its noninvasiveness allows maintaining the physiologic state of embryos. Herein, we show the use of M-mode ultrasound to measure the heart rates of embryos at E18.5 in utero.

في قياس الرحم من معدل ضربات القلب في الماوس بواسطة موسع M-وضع ضربات القلب

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease ...

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the world&#8217;s 2.9 million neonatal deaths. Animal models of HIE have contributed to the understanding of the pathophysiology in HIE, and have highlighted the dynamic process that occur in brain injury due to perinatal asphyxia. Thus, animal studies have suggested a time-window for post-insult treatment strategies. Hypothermia has been tested as a treatment for HIE in pdiglet models and subsequently proven effective in clinical trials. Variations of the model have been applied in the study of adjunctive neuroprotective methods and piglet studies of xenon and melatonin have led to clinical phase I and II trials1,2. The piglet HIE model is further used for neonatal resuscitation- and hemodynamic studies as well as in investigations of cerebral hypoxia on a cellular level. However, it is a technically challenging model and variations in the protocol may result in either too mild or too severe brain injury. In this article, we demonstrate the technical procedures necessary for establishing a stable piglet model of neonatal HIE. First, the newborn piglet (&#60; 24 hr old, median weight 1500 g) is anesthetized, intubated, and monitored in a setup comparable to that found in a neonatal intensive care unit. Global hypoxia-ischemia is induced by lowering the inspiratory oxygen fraction to achieve global hypoxia, ischemia through hypotension and a flat trace amplitude integrated EEG (aEEG) indicative of cerebral hypoxia. Survival is promoted by adjusting oxygenation according to the aEEG response and blood pressure. Brain injury is quantified by histopathology and magnetic resonance imaging after 72 hr.

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 ...

Simultaneous PET/MRI Imaging During Mouse Cerebral Hypoxia-ischemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics disturbance in affected cells. Diffusion weighted magnetic resonance imaging (MRI) identifies regions that are damaged, potentially irreversibly, by hypoxia-ischemia. Alterations in glucose utilization in the affected tissue may be detectable by positron emission tomography (PET) imaging of 2-deoxy-2-(18F)fluoro-&#7429;-glucose ([18F]FDG) uptake. Due to the rapid and variable nature of injury in this animal model, acquisition of both modes of data must be performed simultaneously in order to meaningfully correlate PET and MRI data. In addition, inter-animal variability in the hypoxic-ischemic injury due to vascular differences limits the ability to analyze multi-modal data and observe changes to a group-wise approach if data is not acquired simultaneously in individual subjects. The method presented here allows one to acquire both diffusion-weighted MRI and [18F]FDG uptake data in the same animal before, during, and after the hypoxic challenge in order to interrogate immediate physiological changes.

The method presented here uses simultaneous positron emission tomography and magnetic resonance imaging. In the cerebral hypoxia-ischemia model, dynamic changes in diffusion and glucose metabolism occur during and after injury. The evolving and irreproducible damage in this model necessitates simultaneous acquisition if meaningful multi-modal imaging data are to be acquired.

في وقت واحد التصوير PET / MRI خلال الماوس الدماغي نقص الأكسجة-نقص التروية

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics ...

A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia

Stroke research has endured many setbacks in translating neuroprotective therapies into clinical practice. In contrast, the real-world therapy (tPA thrombolysis) rarely produces benefits in mechanical occlusion-based experimental models, which dominate preclinical stroke research. This split between the bench and bedside suggests the need to employ tPA-responsive models in preclinical stroke research. To this end, a simple and tPA-reactive thrombotic stroke model is invented and described here. This model consists of transient occlusion of the unilateral common carotid artery and delivery of 7.5% oxygen through a face mask in adult mice for 30 min, while maintaining the animal rectal temperature at 37.5 &#177; 0.5 &#176;C. Although reversible ligation of the unilateral carotid artery or hypoxia each suppressed cerebral blood flow only transiently, the combination of both insults caused lasting reperfusion deficits, fibrin and platelet deposition, and large infarct in the middle cerebral artery-supplied territory. Importantly, tail-vein injection of recombinant tPA at 0.5, 1, or 4 hr post-tHI (10 mg/kg) provided time-dependent reduction of the mortality rate and infarct size. This new stroke model is simple and can be standardized across laboratories to compare experimental results. Further, it induces thrombosis without craniectomy or introducing pre-formed emboli. Given these unique merits, the tHI model is a useful addition to the repertoire of preclinical stroke research.

Thromboembolic stroke models are vital tools for optimizing the recanalization therapy. Here we report a murine thrombotic stroke model based on transient cerebral hypoxic-ischemic (tHI) insult, which triggers thrombosis and infarction, and responds favorably to tissue plasminogen activator (tPA)-mediated fibrinolysis in a therapeutic window similar to those in stroke patients.

A السكتة الدماغية نموذج الخثاري بناء على عابر الدماغي نقص الأكسجة-نقص التروية

Stroke research has endured many setbacks in translating neuroprotective therapies into clinical practice. In contrast, the real-world therapy (tPA ...

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 Model to Simulate Clinically Relevant Hypoxia in Humans

In case of apnea, arterial partial pressure of oxygen (pO2) decreases, while partial pressure of carbon dioxide (pCO2) increases. To avoid damage to hypoxia sensitive organs such as the brain, compensatory circulatory mechanisms help to maintain an adequate oxygen supply. This is mainly achieved by increased cerebral blood flow. Intermittent hypoxia is a commonly seen phenomenon in patients with obstructive sleep apnea. Acute airway obstruction can also result in hypoxia and hypercapnia. Until now, no adequate model has been established to simulate these dynamics in humans. Previous investigations focusing on human hypoxia used inhaled hypoxic gas mixtures. However, the resulting hypoxia was combined with hyperventilation and is therefore more representative of high altitude environments than of apnea. Furthermore, the transferability of previously performed animal experiments to humans is limited and the pathophysiological background of apnea induced physiological changes is poorly understood. In this study, healthy human apneic divers were utilized to mimic clinically relevant hypoxia and hypercapnia during apnea. Additionally, pulse-oximetry and Near Infrared Spectroscopy (NIRS) were used to evaluate changes in cerebral and peripheral oxygen saturation before, during, and after apnea.

Hypoxia simulation in humans has usually been performed by inhaling hypoxic gas mixtures. For this study, apneic divers were used to simulate dynamic hypoxia in humans. Additionally, physiological changes in desaturation and re-saturation kinetics were evaluated with non-invasive tools such as Near-Infrared-Spectroscopy (NIRS) and peripheral oxygenation saturation (SpO2).

نموذج لمحاكاة نقص الأكسجة ذات الصلة سريريا في البشر

In case of apnea, arterial partial pressure of oxygen (pO2) decreases, while partial pressure of carbon dioxide (pCO2) increases. To avoid damage to ...