Yazarlar DHMRI CLAS hayvan bakım personelinin güçlü bir destek kabul etmek istiyorum, Tesis Müdürü Sayın Daniel Peralta ve katılan veteriner Dr. Glicerio Ignacio, DVM MRCVS.

Butun hayvan prosedürleri, protokol 14-017 altında, Kuzey Karolina Araştırma Campus (NCRC) üzerinde yer alan David H. Murdock Araştırma Enstitüsü IACUC onaylı edildi, diyabet ve prediyabet / insülin direnci ve terapötik etkinliğinin bir insan olmayan üstün ırk modelinin karakterizasyonu geliştirmek insülin duyarlılığı ve metabolik fonksiyonu. 1. Hayvan Seçimi ve Çalışma Hazırlık <ol><li> aylık gıda alımı ve vücut ağırlığı kayıtlarına dayanarak Seç diyet ve kilo istikrarlı hayvanlar. Not: yiyecek alımı stabilize kadar iştah son bir azalma sergiledi hayvanlar, özelliği olmamalıdır. <ol><li> olgun hayvanlar için (&gt; 5-6 yıl), bedeni ay üst üste arasındaki ağırlıklar ilk maymun incelenmesi ve ağırlık dramatik değişim için metabolik durumunu değiştirmeden başka nedenler ekarte etmeden% 10&#39;dan fazla farklılık hayvanları seçmeyin. </li></ol></li><li> glukoz, insülin ve C-peptidleri içinde analitler, proteaz inhibitörü (Aprotinin ve DPP4I) K 2 EDTA numune toplama tüpleri hazırlayın. Not: DPP4I + Aprotinin kokteyli proteaz inhibisyonu geniş spektrumlu bir sağlar, bu ek analitler (glukagon, GLP-1) için gerekli olmalıdır. Ek analitler toplanan olmadığını durumunda, kan tüpleri Numune alımında tutarlılığı korumak için aynı şekilde hazırlanmalıdır. Bu yöntem için doğrulanmamıştır tahliller için alınan örnekler üreticinin önerilerine göre, ayrı ayrı toplanmalıdır. <ol><li> DPP4I 10 ml Aprotinin, liyofilize 100 mg karıştırılarak proteaz inhibitör kokteyli hazırlanır. Toplanan kan, her mililitre için her kan tüpüne Aprotinin + DPP4I karışımı 10 ul ekle. </li><li> olası tahsilat fazlalık her tüpe proteaz inhibitör kokteyli ilave 10 ul ekle. kullanılana kadar -20 ° C&#39;de muamele kan tüpleri saklayın. procedu sırasında ıslak buz üzerinde tüpler tutunyeniden 4 ° C &#39;de dönel. </li><li> Standart serum kimyası analiz için başlangıçta bir kan örneği toplamak için bir serum ayırıcı tüp kullanın. Tam kan sayımı (CBC) için, proteaz inhibitörleri olmadan standart K 2 EDTA kullanın. kan örnekleri sonrasında kısım plazma ve serum kullanın cryovials aşağı bükülmüş edilmiştir. </li><li> hayvan tanımlama, tarih, prosedür, timepoint ve örnek hacmi ile uygun kan tüpleri ve cryovials etiketleyin. Uygun deney için plazmada analiz (ler) ile birlikte cryovials etiketleyin. </li></ol></li><li> normal tuzlu su içinde 250 ml&#39;lik bir torbaya ml heparin 1.000 USP birim 0.15 ml enjekte ile heparinize edilmiş tuz akışı hazırlayın. 0.06 mg heparin / ml bir solüsyon elde edilir. örnekler arasında temizlenmesi için bir tuzlu su kilide Bu çözeltiye 60 ml - 40 çizin. sırasıyla infüzyon önce ve sonra dekstroz infüzyonu noktası yıkama için ayrı şırıngalar içine ilave 1 mL ve 5 mL çizin. </li></ol> 2.Hayvan Sedasyon ve Hazırlık <ol><li> hayvanın kafesinden işlem öncesi en az 14 saat ve en fazla 18 saat yiyecek çıkarın. NOT: Bu hayvanların glisemik değerleri herhangi bir postprandial varyasyonu önlemek için prosedür için aç olması önemlidir. Ayrıca anestezi sırasında yetersizliği ve mide içeriğinin aspirasyonu önlemek için bir önlemdir. </li><li> Genel anestezik olarak kas içinden verilen ketamin ile IVGTT prosedür süresince oturaklı hayvanlar, 10 mg / kg. İşlem sırasında, gerektiğinde, 30 dakika ya da 20 - aralıklarla - (10 mg / kg ve 5), ek ketamin yönetme. <ol><li> sedasyon hayvan tartılır. ısıtılmış bir prosedür masaya yanal yatar pozisyonda hayvan yerleştirin. </li><li> Hayvan anestezi istikrarlı bir düzlemde sağlamak için klinik parametrelerin her 15 ila 20 dakika izleyin. Bir pulse oksimetre ile - (200 bpm 100) ve DPT 2 (&gt;% 92) kalp hızını ölçün. (- 50 nefes / dk 20) ​​w solunum hızını ölçmek<html> Bir kronometre i, görsel ya da on beş saniye boyunca elle solunumda sayarak ve dört ile çarpılarak. Tedbir sıcaklığı (&gt; 97 ° F) rektal. sakız ve dudakların (nemli, pembe) etrafında mukoz membran rengini izleyin. </li></ol></li><li> İki kanül siteleri hazırlayın. Kateter eklenecek ilgi alanından saç kesme ve alternatif klorheksidin scrubs ve% 70 alkol ile tüm bölgeyi sterilize etmek için saç makasları kullanın. <ol><li> Sol veya sağ sefalik veya safen venlerin bölgesinde bir kateter yerleştirilir ve üç yollu ile (mg heparin / ml 0.06) bir heparin tuzlu floş ekleyin. Bu örnekleme kan alımı sitesidir. </li><li> Başka bir bacak ikinci kateter yerleştirin veya sefalik veya safen venlerin bölgede kol ve bir bağlantı noktası ekleyin. dekstroz infüzyon için bu siteyi kullanın. önce dekstroz infüzyon için kanül patent tutmak için, küçük heparinize salin 1 ml floş kullanın. </li></ol></li></ol>e_title &quot;&gt; 3. İVGTT Prosedürü Not: IVGTT Prosedür 8 kan alımı numune zaman noktaları (Tablo 1) oluşur. <ol><li> Bazal örneği alın ve açlık kan şekeri düzeyini ölçmek için bir el glukometre kullanın. hayvanın genel sağlığını değerlendirmek için bir CBC standart kimya analizi için bir serum numunesi, aynı zamanda, bütün bir kan örneği alın. Imalatçının talimatlarına 8, 9 uyarınca makaklarda kullanılmak için kabul edilmiş bir kit kullanılarak taban numuneden glikoz ve insülin seviyelerini analiz etmek için plazma numuneleri toplamak. Not: Bu, önceden kanülün ölü boşluk kalan kan veya heparin çıkarmak için kan toplanması için numune alma kanül alınan 0.5 ml&#39;lik bir ön çekmesi önemlidir. </li><li> Bazal örnek alınmasını takiben, dekstroz infüzyonu portuna 30 saniye içinde% 50 dekstroz (250 mg / kg) infüzyonu. Not: yüksek doz modelleri (500 mg / kg), kullanılabilirgerçi doz uzunlamasına karşılaştırmalar yapmak amacıyla prosedürler arasında sabit olmalıdır. <ol><li> Limanda kalmadı dekstroz olduğundan emin olmak için heparinli serum fizyolojik 5 ml infüzyon portu yıkayın. infüzyon sonu T0 olduğunu. infüzyon arta kalan dekstroz sonraki kan örnekleri kontamine olabilir teknisyen, eldiven yerine var. </li></ol></li><li> sonrası ilk infüzyon örnek zaman noktası T5 dakika T7 dakika, T10 dakika, T15 dakika, T20 min ardından dekstroz infüzyon sonunda, gelen, T3 dk ve son örnek zaman noktası T30 min yer almaktadır. Başlangıç ​​numunesi ile deney glikoz ve insülin düzeyleri her bir zaman noktasında plazmanın kazan (adım 3.1). <ol><li> zaman noktasında dk T3, bir kez daha kan şekeri seviyesini kontrol etmek için el glukometre kullanın. NOT: Başlangıçta Glucometer okumalar ve T3 dekstroz infüzyon onaylamak için sadece. zaman noktasında dk T3 kan glukoz düzeyi ~ 100 mg / dl daha yüksek eş olmalıdıraçlık bazal plazma glukoz düzeyine mpared. </li></ol></li></ol> 4. Hayvan Kurtarma ve Örnek İşleme <ol><li> kanüller çıkarın ve zaman noktası dk T30 sonra hemostaz için kateterize sitelere basınç uygulayın. o kendine geldi ve oturmuş kadar hayvan izleyin. hayvan kez Teklif besleme tamamen iyileşti edilir. </li><li> Hemen buz üzerine K 2, EDTA tüpleri içine her bir tam kan örneği yerleştirin. toplama, 10 dakika içinde 4 ° C arasındaki bir sıcaklıkta 3000 rpm&#39;de santrifüj. analize kadar -80 ° C &#39;de cryovials, dondurulması ve depolanması içine kısım plazma numuneleri. <ol><li> Standart serum kimyası analizleri için serum tüpü içine, kan az 20 dakika ve oda sıcaklığında 3000 rpm&#39;de santrifüjlemeden önce en fazla 30 dakika süreyle, oda sıcaklığında bekletin. toplama 48 saat içerisinde deneye tabi tutulana kadar, serum örnekleri dondurun. NOT: olarak kadar CBC analizi için toplanan buzdolabında tam kan örnekleritoplama 24 saat içinde sayed. </li></ol></li></ol> 5. Veri Tedavisi <ol><li> Plazma insülin ve glikoz eğrisi oluşturulması başlangıçta, 16, 17, yukarıda glukoz değerleri için doğal log A eğiminden glukoz klirensi oranı K belirler. Not: Sağlıklı bir NSP sağlıklı hayvan genellikle 30 dakika içinde, başlangıç glukoz değerlerine geri döner olarak, genellikle daha büyük bir 2 veya daha fazla, iyi 1&#39;in üzerinde bir glükoz klirensi oranı K olması beklenebilir. insülin üretimi devre dışı bırakır gibi, glikoz temizlenme oranı K 1&#39;in altına düşen, daha önemli ölçüde düşecek. </li><li> T30 16, 17 vasıtasıyla, zaman noktaları arasında, her çizgi parçasının eğri altında kalan alan temsil yamuk alanının toplamları toplamı olarak, bir bütün olarak AUC hesaplayın. Not: Geleneksel olarak, işlemin ilk on dakika AUC akut olarak kabul edilirprosedürün son 20 dk AUC geç insülin yanıtı (LIR) kabul edilir ise insülin cevabı e, (AIR) glukoz. Hayvan daha dismetabolik hale geldikçe, insülin AUC insülin duyarsızlığı artırmak için tazminat yansıtan artacaktır. Aşikar diyabet hayvan geçişleri gibi, ancak, AUC AIR tarafından yakalanan, akut insülin aşamasında genellikle başlangıçta, azalacaktır. </li></ol>

<ol>
	<li>Bergman, R., Phillips, L., Cobelli, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiologic+evaluation+of+factors+controlling+glucose+tolerance+in+man.">Physiologic evaluation of factors controlling glucose tolerance in man.</a> J. Clin. Invest. 68, 1456-1457 (1981).</li><li>Bergman, R., Prager, R., Volund, A., Olefsky, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Equivalence+of+the+insulin+sensitivity+index+in+man+derived+by+the+minimal+model+and+the+euglycemic+glucose+clamp.">Equivalence of the insulin sensitivity index in man derived by the minimal model and the euglycemic glucose clamp.</a> J. Clin. Invest. 79, 790-800 (1987).</li><li>Hovorka, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partitioning+glucose+distribution/transport,+disposal,+and+endogenous+production+during+IVGTT.">Partitioning glucose distribution/transport, disposal, and endogenous production during IVGTT.</a> Am. J. Physiol. Endocrinol. Metab. 282, E992-E1007 (2002).</li><li>Salinari, S., Guidone, C., Bertuzzi, A., Manco, M., Asnaghi, S., Mingrone, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=First-phase+insulin+secretion+restoration+and+differential+response+to+glucose+load+depending+on+the+route+of+administration+in+type+2+diabetic+subjects+after+beriatric+surgery.">First-phase insulin secretion restoration and differential response to glucose load depending on the route of administration in type 2 diabetic subjects after beriatric surgery.</a> Diabetes Care. 32 (3), 375-380 (2009).</li><li>Roden, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Clinical Diabetes Research: Methods and Techniques. , (2007).</li><li>Cobelli, C., Pacini, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insulin+secretion+and+hepatic+extraction+in+humans+by+minimal+modeling+of+c-peptide+and+insulin+kinetics.">Insulin secretion and hepatic extraction in humans by minimal modeling of c-peptide and insulin kinetics.</a> Diabetes. 37, 223-231 (1988).</li><li>Lorenzo, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disposition+index,+glucose+effectiveness,+and+conversion+to+type+2+diabetes:+the+insulin+resistance+atherosclerosis+study.">Disposition index, glucose effectiveness, and conversion to type 2 diabetes: the insulin resistance atherosclerosis study.</a> Diabetes Care. 33, 2098-2103 (2010).</li><li>Hansen, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+and+treatment+of+type+2+diabetes+in+nonhuman+primates.">Investigation and treatment of type 2 diabetes in nonhuman primates.</a> Methods Mol Biol. 933, 177-185 (2012).</li><li>Hansen, B. C., Bodkin, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standardization+of+IVGTT.+Importance+of+method+used+to+calculate+glucose+disappearance.">Standardization of IVGTT. Importance of method used to calculate glucose disappearance.</a> Diabetes Care. 16 (5), 847 (1993).</li><li>Hardwood, J. H., Listrani, P., Wagner, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonhuman+primates+and+other+animal+models+in+diabetes+research.">Nonhuman primates and other animal models in diabetes research.</a> J Diabetes Sci Tech. 3, 503-514 (2012).</li><li>De Koning, E. J., Bodkin, N. L., Hansen, B. C., Clark, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diabetes+mellitus+in+Macaca+mulatta+monkeys+is+characterized+by+islet+amyloidosis+and+reduction+in+beta-cell+population.">Diabetes mellitus in Macaca mulatta monkeys is characterized by islet amyloidosis and reduction in beta-cell population.</a> Diabetologia. 36, 378-384 (1993).</li><li>Letiexhe, M. R., Scheen, A. J., Gerard, P. L., Desaive, C., Lefebvre, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insulin+secretion,+clearance+and+action+before+and+after+gastroplasty+in+severely+obese+subjects.">Insulin secretion, clearance and action before and after gastroplasty in severely obese subjects.</a> Int J Obes Relat Metab Disord. 18, 295-300 (1994).</li><li>Letiexhe, M. R., Scheen, A. J., Gerard, P. L., Desaive, C., Lefebvre, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postgastroplasty+recovery+of+ideal+body+weight+normalizes+glucose+and+insulin+metabolism+in+obese+women.">Postgastroplasty recovery of ideal body weight normalizes glucose and insulin metabolism in obese women.</a> J Clin Endocrinol Metab. 80, 364-369 (1995).</li><li>Kim, S. H., Abbasi, F., Chu, J. W., McLaughlin, T. L., Lamendola, C., Polonsky, K. S., Reaven, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rosiglitazone+reduces+glucose-stimulated+insulin+secretion+rate+and+increases+insulin+clearance+in+nondiabetic,+insulin-resistant+individuals.">Rosiglitazone reduces glucose-stimulated insulin secretion rate and increases insulin clearance in nondiabetic, insulin-resistant individuals.</a> Diabetes. 54, 2447-2452 (2005).</li><li>Toffolo, G., Breda, E., Cavaghan, M. K., Ehrmann, D. A., Polonsky, K. S., Cobelli, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+indexes+of+beta-cell+function+during+graded+up+and+down+glucose+infusion+from+C-peptide+minimal+models.">Quantitative indexes of beta-cell function during graded up and down glucose infusion from C-peptide minimal models.</a> Am J Physiol Endocrinol Metab. 280, E2-E10 (2001).</li><li>Wang, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+beta-cell+insulin+secretory+function+using+a+graded+glucose-infusion+with+C-peptide+deconvolution+in+dysmetabolic,+and+diabetic+cynomolgus+monkeys.">Quantification of beta-cell insulin secretory function using a graded glucose-infusion with C-peptide deconvolution in dysmetabolic, and diabetic cynomolgus monkeys.</a> Diabetology and Metabolic Syn. 5, 40 (2013).</li><li>Xiao, Y. F., Wang, B., Wang, X., Du, F., Benzinou, M., Wang, Y. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Xylazine-induced+reduction+of+tissue+sensitivity+to+insulin+leads+to+acute+hyperglycemia+in+diabetic+and+normoglycemic+monkeys.">Xylazine-induced reduction of tissue sensitivity to insulin leads to acute hyperglycemia in diabetic and normoglycemic monkeys.</a> Anesthesiology. 13 (33), (2013).</li><li>Porte, D., Kahn, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=&#946;-cell+dysfunction+and+failure+in+type+2+diabetes+potential+mechanisms.">&#946;-cell dysfunction and failure in type 2 diabetes potential mechanisms.</a> Diabetes. 50, S160-S163 (2001).</li><li>DeFronzo, R. A., Tobin, J. D., Andres, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glucose+clamp+technique:+a+method+for+quantifying+insulin+secretion+and+resistance.">Glucose clamp technique: a method for quantifying insulin secretion and resistance.</a> American Journal of Physiology. 237 (3), G214-G223 (1979).</li><li>Ferrannini, E., Gastaldelli, A., Miyazaki, Y., Matsuda, M., Mari, A., DeFronzo, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=&#946;-cell+function+in+subjects+spanning+the+range+from+normal+glucose+tolerance+to+overt+diabetes:+a+new+analysis.">&#946;-cell function in subjects spanning the range from normal glucose tolerance to overt diabetes: a new analysis.</a> J Clin Endocrinol Metab. 90 (1), 493-500 (2005).</li><li>Vaughan, K. L., Szarowicz, M. D., Herbert, R. L., Mattison, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+anesthesia+protocols+for+intravenous+glucose+tolerance+testing+in+rhesus+monkeys.">Comparison of anesthesia protocols for intravenous glucose tolerance testing in rhesus monkeys.</a> J Med Primatol. 43, 162-168 (2014).</li><li>Kemnitz, J. W., Kraemer, G. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+glucoregulation+in+rhesus+monkeys+sedated+with+ketamine.">Assessment of glucoregulation in rhesus monkeys sedated with ketamine.</a> American Journal of Primatology. 3, 201-210 (1982).</li><li>Dutton, C. J., Parvin, C. A., Gronowski, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+glycated+hemoglobin+percentages+for+use+in+the+diagnosis+and+monitoring+of+diabetes+mellitus+in+nonhuman+primates.">Measurement of glycated hemoglobin percentages for use in the diagnosis and monitoring of diabetes mellitus in nonhuman primates.</a> Am J Vet Res. 64, 562-568 (2003).</li><li>Rai, V., Iyer, U., Mani, I., Mani, U. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serum+biochemical+changes+in+insulin+dependent+and+non-insulin+dependent+diabetes+mellitus+and+their+role+in+the+development+of+secondary+complications.">Serum biochemical changes in insulin dependent and non-insulin dependent diabetes mellitus and their role in the development of secondary complications.</a> Int J Diab Dev Countries. 17, 33-37 (1997).</li><li>Shirasaki, Y., Yoshioka, N., Kanazawa, K., Maekawa, T., Horikawa, T., Hayashi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+physical+restraint+on+glucose+tolerance+in+cynomolgus+monkeys.">Effect of physical restraint on glucose tolerance in cynomolgus monkeys.</a> J Med Primatol. 42, 165-168 (2013).</li></ol>

Yazarlar metabolik hastalık alanında aktif bir sözleşmeli araştırma kuruluşuna (Taç Bioscience) ile bağlıdırlar.

İVGTT tahlil itibaren, açlık kan şekeri ve insülin seviyesinin elde edilir. Vücut ağırlığı 5, 12, 13 dayalı tek bir dekstroz infüzyon ile glukoz ile uyarılmış insülin salınımı kapasitesini değerlendirir ve hayvanın kapasitesinin değerlendirilmesini sağlar insülin bırakın ve başlangıca doğru yükselmiş glukoz düzeyi döndürür. Bu normal bir glukoz ve insülin düzeyi, sağlıklı kontrol, Normogliseminin ile hiperinsülinemik dismetabolik hayvan veya bir hiperglisemik insülin...

IVGTT rutin farklı metabolik Devletleri 5 7 insanlarda β-hücresi fonksiyonunu belirlemek için kullanılan uygun bir işlevsel tahlildir. T2D hayvan modellerinde, bu da iyi metabolik hastalık ilerlemesini gösteren hayvanlar karakterize etmek için bir araç olarak kabul edilmektedir bir dismetabolik hiperglisemik hal 8, 9 sağlıklı. T2D en yakın hayvan modeli, rhesus ve sinomolgus maymunları önemli örneklerdir olan insan olmayan primatlarda (NHPs), gösterilmektedir. Bu hayvanlar doğal insanlarda 10 gibi yaş ve sıklığı katkıda obezite aynı risk faktörleri ile T2D gelişir. Ayrıca, orada da benzer bir hastalık ilerlemesi ve dismetabolik hastalığı 11 ilerledikçe pankreas patoloji amiloid birikimi gösteren. İşte bu hayvanlarda metabolik durumunun bizim koloni karakterizasyonu bir parçası olarak NHPs bir İVGTT gerçekleştirme standart yönteme rapor. Bu yöntem, birtüketen diğer, daha zaman ve pahalı teknikler 2 göreli gerçekleştirmek kolay. IVGTT hızlı ve sık sık hayvanların büyük bir koloni karakterize etmek için yararlıdır. Glike hemoglobin (HbA1C) düzeyi ile dikkate alındığında, hayvanın beslenme ve gıda alımı tarih, yanı sıra yüzde yağsız kütle ve vücut yağ, İVGTT normalde aşikar diyabet 6 doğru bir hayvanın metabolik durumunu ve ilerlemesini karakterize etmek için yeterlidir 8. HbA1C üç ay önceki altı hafta içinde glikoz seviyelerinin güvenilir bir ölçümünü sağlayan bir kırmızı kan hücresinin ömrü boyunca ortalama glisemik düzeyini temsil eder. IVGTT&#39;nin oruç bazal kan örneğinden ölçüldüğünde, bu değer prosedürleri arasında aylarda glisemik kontrol içine bir pencere sağlar. hayvan son IVGTT&#39;nin beri diyabetik için dismetabolik dan geçiş ise, önceki değerinden çok daha yüksek bir HbA1C değeri işaret ederGeçiş yakında başladı son IVGTT&#39;nin sonra ise, yakın önceki değere bir HbA1C değeri sadece son zamanlarda geçişini olduğunu işaret eder. Genel olarak, rhesus makak içinde, HbA1C% 6 kötü glisemik kontrolü 10, 23 anormal kabul edilir ve işaret edilir daha fazla değer verir. Glisemi düzeyleri bir bütün olarak hayvan davranışı ve genel sağlığı kapsamında değerlendirilmelidir. Diyabetik makakları - sergi hyperphagia, polidipsi, poliüri ve - insanlar gibi. Hayvanların grup konut dismetabolik ve diyabetik maymunlar için gerekli bu göstergeler ve bireysel bakım ölçümü için önemli zorluklar sağlar. Bu, tek başına fazla kişisel bakım sağlanabilir sırayla hayvanları konut tavsiye ve daha kolay bir şekilde maymun sağlığı davranışsal belirteçleri 8 izlenmelidir. Buna ek olarak, diyabetik makakları kilo kaybı, yanı sıra artan yüksek bir lipid profili (sergileyecekkolesterol, hipertrigliseridemi) ve serum kimyasında rahatsız mineral metabolizması. Bu organlara zarar metabolik bozukluk / diyabet ilerleyen komplikasyonları genellikle, ve glisemik, lipid ve mineral dengesizlikleri 9, 11, 18 ortak belirleyicileri olabilir 24 gibi, karaciğer ve serum kimyasında böbrek fonksiyonunun belirteçleri ölçmek önemlidir . Bu yöntemi kullanırken bir maymunun hayat belirli değere sahip üzerine, tarihi değerleri çoklu sık karakterizasyonu oluşturulan. Böyle bir glikoz kelepçe veya kademeli glukoz infüzyon (GGI) gibi diğer işlemler, tam bir hayvanın sağlığını değerlendirmek için gerekli olup olmadığını kendi geçmişi kullanılamaz olduğunda, ilk karakterizasyonu üzerine yaygın olduğunu. her üç ayda bir bir sıklıkta bir temel oluşturulmuştur Ancak, bir kez tekrarlanan IVGTT&#39;ler, operasyondan bir hayvanın ilerlemesini takip etmek için, normal olarak yeterlidir. hayvanlar üzerinde birden fazla çalışmalar alındı ​​zaman bu özellikle önemlidirtakvim yılı metabolik durumuna dayalı. sağlık bir seferde yıl nispeten istikrarlı kalabilir sırasında bir hayvanın metabolik durumu kötüleşir zaman, insülin direnci ve glukoz intoleransı dramatik bir artış çok hızlı oluşabilir. HbA1C değerleri üç ay arayla planlanan prosedürleri arasında hayvan sağlık durumunun azalması ya da bazı gelişmeler enterpolasyon için izin verir. Bu nedenle, bu yöntem, doğal ömrü boyunca birden fazla uzunlamasına çalışmalarda kullanılan hayvanların vasıflandırılması için idealdir.

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characterization of metabolic status in nonhuman primates with the intravenous glucose tolerance test

İntravenöz glikoz tolerans testi (IVGTT) glikoz dengesinin karakterizasyonu önemli bir rol oynar. serum biyokimyasal profilleri, hem beslenen kan glikoz seviyelerinin kapsayıcı ve açlık durumunda, HbA1c, insülin düzeyleri, diyet klinik öykü, vücut kompozisyonu ve vücut ağırlığı durumu, normal ve anormal glisemik kontrolün değerlendirilmesi ile birlikte alındığında yapılabilir . Bir IVGTT&#39;nin yorumlanması dekstroz meydan ilişkin zamanla kan şekeri ve insülin düzeylerinde değişiklikler ölçümü ile yapılır. Dikkate alınması gereken kritik bileşenleri şunlardır: T0 (glikoz infüzyon sonu), ilk 20 dakika içinde hızlı glikoz temizlenmesi eğiminden elde edilen glukoz temizlenme oranı K (T20 T1), zamana göre ulaşılan zirve glikoz ve insülin düzeyleri glikoz taban dönmek için, ve eğri (AUC) altındaki alan. Bu İVGTT önlemler sağlıklı bir t glikoz homeostasis hareket ettikçe karakteristik değişiklikler gösteriroa hastalıklı metabolik durumu 5. Bu yazıda gelen insanlarda Tip II diyabet (T2D) en alakalı hayvan modeli ve İVGTT ve bu hayvanların klinik profilleri insan dışı primatlarda (Rhesus ve sinomolgus makakları), karakterizasyonu anlatacağım bir obez dismetabolik için, sağlıklı yalın, ve T2D devlet 8, 10, 11.

Şekil 1 &#39;de gösterilen sonuçlar, bir 30 dakika IVGTT&#39;nin boyunca, olgun, sağlıklı ve diyabetik sinomolgus makaklar tipik glükoz ve insülin eğrilerinin gösterim bulunmaktadır. Sağlıklı ve ilerlemiş diyabetik maymunlar elde edilen veriler metabolik karakterizasyonu aralığının her iki uç ucundan hayvanlar arasında belirgin farklılıklar kontrast için gösterilmiştir. Bu İVGTT protokol benzer sonuçlar ile rhesus makak yazarlar tarafından ba...

Watch this Scientific Journal Video about Characterization of Metabolic Status in Nonhuman Primates with the Intravenous Glucose Tolerance Test at JoVE.com

Characterization of Metabolic Status in Nonhuman Primates with the Intravenous Glucose Tolerance Test

Bu protokolün amacı, insan dışı primatlarda glisemik kontrolü değerlendirmek ve dismetabolik sağlıklı onların metabolik durumunu değerlendirmek için intravenöz glikoz tolerans testi (IVGTT&#39;ler) gerçekleştirmek için standart bir yöntem sunmaktır.

İntravenöz Glikoz Tolerans Testi ile insan olmayan primatlarda Metabolik Durumu Karakterizasyonu

The intravenous glucose tolerance test (IVGTT) plays a key role in the characterization of glucose homeostasis. When taken together with serum biochemical ...

characterization-metabolic-status-nonhuman-primates-with-intravenous

Research

JoVE Journal

Medicine

10.8K Görüntüleme.  Crown Bioscience.  Bu protokolün amacı, insan dışı primatlarda glisemik kontrolü değerlendirmek ve dismetabolik sağlıklı onların metabolik durumunu değerlendirmek için intravenöz glikoz tolerans testi (IVGTT'ler) gerçekleştirmek için standart bir yöntem sunmaktır. 

Video: Characterization of Metabolic Status in Nonhuman Primates with the Intravenous Glucose Tolerance Test

Making Sense of Listening: The IMAP Test Battery

The ability to hear is only the first step towards making sense of the range of information contained in an auditory signal. Of equal importance are the abilities to extract and use the information encoded in the auditory signal. We refer to these as listening skills (or auditory processing AP). Deficits in these skills are associated with delayed language and literacy development, though the nature of the relevant deficits and their causal connection with these delays is hotly debated.
When a child is referred to a health professional with normal hearing and unexplained difficulties in listening, or associated delays in language or literacy development, they should ideally be assessed with a combination of psychoacoustic (AP) tests, suitable for children and for use in a clinic, together with cognitive tests to measure attention, working memory, IQ, and language skills. Such a detailed examination needs to be relatively short and within the technical capability of any suitably qualified professional. Current tests for the presence of AP deficits tend to be poorly constructed and inadequately validated within the normal population. They have little or no reference to the presenting symptoms of the child, and typically include a linguistic component. Poor performance may thus reflect problems with language rather than with AP. To assist in the assessment of children with listening difficulties, pediatric audiologists need a single, standardized child-appropriate test battery based on the use of language-free stimuli.
We present the IMAP test battery which was developed at the MRC Institute of Hearing Research to supplement tests currently used to investigate cases of suspected AP deficits. IMAP assesses a range of relevant auditory and cognitive skills and takes about one hour to complete. It has been standardized in 1500 normally-hearing children from across the UK, aged 6-11 years. Since its development, it has been successfully used in a number of large scale studies both in the UK and the USA. IMAP provides measures for separating out sensory from cognitive contributions to hearing. It further limits confounds due to procedural effects by presenting tests in a child-friendly game-format. Stimulus-generation, management of test protocols and control of test presentation is mediated by the IHR-STAR software platform. This provides a standardized methodology for a range of applications and ensures replicable procedures across testers. IHR-STAR provides a flexible, user-programmable environment that currently has additional applications for hearing screening, mapping cochlear implant electrodes, and academic research or teaching.

A test battery (IMAP) for performing an in-depth assessment of auditory and cognitive abilities contributing to listening skills is described. It is quick to administer, child-friendly and free from linguistic confounds. Stimulus generation and protocol management are controlled via a software platform (IHR-STAR) to ensure replicable procedures.

Dinlemede Making Sense IMAP Test Batarya:

The ability to hear is only the first step towards making sense of the range of information contained in an auditory signal. Of equal importance are the ...

Tıp

Delivery of Therapeutic Agents Through Intracerebroventricular (ICV) and Intravenous (IV) Injection in Mice

Despite the protective role that blood brain barrier plays in shielding the brain, it limits the access to the central nervous system (CNS) which most often results in failure of potential therapeutics designed for neurodegenerative disorders 1,2. Neurodegenerative diseases such as Spinal Muscular Atrophy (SMA), in which the lower motor neurons are affected, can benefit greatly from introducing the therapeutic agents into the CNS. The purpose of this video is to demonstrate two different injection paradigms to deliver therapeutic materials into neonatal mice soon after birth. One of these methods is injecting directly into cerebral lateral ventricles (Intracerebroventricular) which results in delivery of materials into the CNS through the cerebrospinal fluid 3,4. The second method is a temporal vein injection (intravenous) that can introduce different therapeutics into the circulatory system, leading to systemic delivery including the CNS 5. Widespread transduction of the CNS is achievable if an appropriate viral vector and viral serotype is utilized. Visualization and utilization of the temporal vein for injection is feasible up to postnatal day 6. However, if the delivered material is intended to reach the CNS, these injections should take place while the blood brain barrier is more permeable due to its immature status, preferably prior to postnatal day 2. The fully developed blood brain barrier greatly limits the effectiveness of intravenous delivery. Both delivery systems are simple and effective once the surgical aptitude is achieved. They do not require any extensive surgical devices and can be performed by a single person. However, these techniques are not without challenges. The small size of postnatal day 2 pups and the subsequent small target areas can make the injections difficult to perform and initially challenging to replicate.

Delivery of Therapeutic Agents Through Intracerebroventricular ICV and Intravenous IV Injection in Mice

This article demonstrates two very different methods of injection: 1) into the brain (intracerebroventricular) and 2) systemic (intravenous) to introduce the therapeutic agents into the central nervous system of neonatal mice.

Fare intraserebroventriküler sayesinde Terapötik Ajanlar teslimi (ICV) ve intravenöz (IV) Enjeksiyon

Despite the protective role that blood brain barrier plays in shielding the brain, it limits the access to the central nervous system (CNS) which most ...

Tilt Testing with Combined Lower Body Negative Pressure: a "Gold Standard" for Measuring Orthostatic Tolerance

Orthostatic tolerance (OT) refers to the ability to maintain cardiovascular stability when upright, against the hydrostatic effects of gravity, and hence to maintain cerebral perfusion and prevent syncope (fainting). Various techniques are available to assess OT and the effects of gravitational stress upon the circulation, typically by reproducing a presyncopal event (near-fainting episode) in a controlled laboratory environment. The time and/or degree of stress required to provoke this response provides the measure of OT. Any technique used to determine OT should: enable distinction between patients with orthostatic intolerance (of various causes) and asymptomatic control subjects; be highly reproducible, enabling evaluation of therapeutic interventions; avoid invasive procedures, which are known to impair OT1.
In the late 1980s head-upright tilt testing was first utilized for diagnosing syncope2. Since then it has been used to assess OT in patients with syncope of unknown cause, as well as in healthy subjects to study postural cardiovascular reflexes2-6. Tilting protocols comprise three categories: passive tilt; passive tilt accompanied by pharmacological provocation; and passive tilt with combined lower body negative pressure (LBNP). However, the effects of tilt testing (and other orthostatic stress testing modalities) are often poorly reproducible, with low sensitivity and specificity to diagnose orthostatic intolerance7.
Typically, a passive tilt includes 20-60 min of orthostatic stress continued until the onset of presyncope in patients2-6. However, the main drawback of this procedure is its inability to invoke presyncope in all individuals undergoing the test, and corresponding low sensitivity8,9. Thus, different methods were explored to increase the orthostatic stress and improve sensitivity.
Pharmacological provocation has been used to increase the orthostatic challenge, for example using isoprenaline4,7,10,11 or sublingual nitrate12,13. However, the main drawback of these approaches are increases in sensitivity at the cost of unacceptable decreases in specificity10,14, with a high positive response rate immediately after administration15. Furthermore, invasive procedures associated with some pharmacological provocations greatly increase the false positive rate1.
Another approach is to combine passive tilt testing with LBNP, providing a stronger orthostatic stress without invasive procedures or drug side-effects, using the technique pioneered by Professor Roger Hainsworth in the 1990s16-18. This approach provokes presyncope in almost all subjects (allowing for symptom recognition in patients with syncope), while discriminating between patients with syncope and healthy controls, with a specificity of 92%, sensitivity of 85%, and repeatability of 1.1&plusmn;0.6 min16,17. This allows not only diagnosis and pathophysiological assessment19-22, but also the evaluation of treatments for orthostatic intolerance due to its high repeatability23-30. For these reasons, we argue this should be the "gold standard" for orthostatic stress testing, and accordingly this will be the method described in this paper.

We describe a "gold standard" for evaluating orthostatic tolerance (OT) using tilt testing with combined lower body negative pressure (LBNP). This can be combined with non-invasive evaluations of cardiovascular reflex control. Normal and abnormal responses are defined.

Kombine Alt Vücut Negatif Basınç ile Tilt Testi: Ortostatik Tolerans Ölçüm için bir &quot;Altın Standart&quot;

Orthostatic tolerance (OT)  refers to the ability to maintain cardiovascular stability when upright,  against the hydrostatic effects of gravity, and ...

Improving Strength, Power, Muscle Aerobic Capacity, and Glucose Tolerance through Short-term Progressive Strength Training Among Elderly People

This protocol describes the simultaneous use of a broad span of methods to examine muscle aerobic capacity, glucose tolerance, strength, and power in elderly people performing short-term resistance training (RET). Supervised progressive resistance training for 1 h three times a week over 8 weeks was performed by RET participants (71&#177;1 years, range 65-80). Compared to a control group without training, the RET showed improvements on the measures used to indicate strength, power, glucose tolerance, and several parameters of muscle aerobic capacity. Strength training was performed in a gym with only robust fitness equipment. An isokinetic dynamometer for knee extensor strength permitted the measurement of concentric, eccentric, and static strength, which increased for the RET group (8-12% post- versus pre-test). The power (rate of force development, RFD) at the initial 0-30 ms also showed an increase for the RET group (52%). A glucose tolerance test with frequent blood glucose measurements showed improvements only for the RET group in terms of blood glucose values after 2 h (14%) and the area under the curve (21%). The blood lipid profile also improved (8%). From muscle biopsy samples prepared using histochemistry, the amount of fiber type IIa increased, and a trend towards a decrease in IIx in the RET group reflected a change to a more oxidative profile in terms of fiber composition. Western blot (to determine the protein content related to the signaling for muscle protein synthesis) showed a rise of 69% in both Akt and mTOR in the RET group; this also showed an increase in mitochondrial proteins for OXPHOS complex II and citrate synthase (both ~30%) and for complex IV (90%), in only the RET group. We demonstrate that this type of progressive resistance training offers various improvements (e.g., strength, power, aerobic capacity, glucose tolerance, and plasma lipid profile).

The effect of short-term resistance training on elderly people was investigated through the simultaneous use of several methods. Compared to a control group, many improvements were seen, including on muscle aerobic capacity, glucose tolerance, strength, power, and muscle quality (i.e., protein involved in cell signaling and muscle fiber type composition).

Yaşlılar Arası Kısa Süreli Progresif Güç Eğitimiyle Mukavemet, Güç, Kas Aerobik Kapasitesi ve Glukoz Toleransının Geliştirilmesi

This protocol describes the simultaneous use of a broad span of methods to examine muscle aerobic capacity, glucose tolerance, strength, and power in ...

Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at the 12 codon (Ptf1acre/+;LSL-KrasG12D/+) mouse strain as a model of human pancreatic cancer. The goal of our current studies is to identify novel metabolic biomarkers of pancreatic cancer progression. We have performed metabolic profiling of urine, feces, blood, and pancreas tissue extracts, as well as histological analyses of the pancreas to stage the cancer progression. The mouse pancreas is not a well-defined solid organ like in humans, but rather is a diffusely distributed soft tissue that is not easily identified by individuals unfamiliar with mouse internal anatomy or by individuals that have little or no experience performing mouse organ dissections. The purpose of this article is to provide a detailed step-wise visual demonstration to guide novices in the removal of the mouse pancreas by dissection. This article should be especially valuable to students and investigators new to research that requires harvesting of the mouse pancreas by dissection for metabolic profiling or histological analyses.

This video article provides a detailed demonstration of the procedures required to successfully remove the pancreas from a mouse by dissection for histological analysis and metabolic profiling.

Fare pankreas diseksiyon histolojik analiz ve metabolik profil oluşturma

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at ...

Studying the Hypothalamic Insulin Signal to Peripheral Glucose Intolerance with a Continuous Drug Infusion System into the Mouse Brain

Insulin regulates systematic metabolism in the hypothalamus and the peripheral insulin response. An inflammatory reaction in peripheral adipose tissues contributes to type 2 diabetes mellitus (T2DM) development and appetite regulation in the hypothalamus. Chemokine CCL5 and C-C chemokine receptor type 5 (CCR5) levels have been suggested to mediate arteriosclerosis and glucose intolerance in type 2 diabetes mellitus (T2DM). In addition, CCL5 plays a neuroendocrine role in the hypothalamus by regulating food intake and body temperature, thus, prompting us to investigate its function in hypothalamic insulin signaling and the regulation of peripheral glucose metabolism.
The micro-osmotic pump brain infusion system is a quick and precise way to manipulate CCL5 function and study its effect in the brain. It also provides a convenient alternative approach to generating a transgenic knockout animal. In this system, CCL5 signaling was blocked by intracerebroventricular (ICV) infusion of its antagonist, MetCCL5, using a micro-osmotic pump. The peripheral glucose metabolism and insulin responsiveness was detected by the Oral Glucose Tolerance Test (OGTT) and Insulin Tolerance Test (ITT). Insulin signaling activity was then analyzed by protein blot from tissue samples derived from the animals.
After 7-14 days of MetCCL5 infusion, the glucose metabolism and insulin responsiveness was impaired in mice, as seen in the results of the OGTT and ITT. The IRS-1 serine302 phosphorylation was increased and the Akt activity was reduced in mice hypothalamic neurons following CCL5 inhibition. Altogether, our data suggest that blocking CCL5 in the mouse brain increases the phosphorylation of IRS-1 S302 and interrupts hypothalamic insulin signaling, leading to a decrease in insulin function in peripheral tissues as well as the impairment of glucose metabolism.

This protocol studies the role of chemokine (C-C motif) ligand 5 (CCL5) in the hypothalamus by delivering an antagonist, MetCCL5, into the mouse brain using a micro-osmotic pump brain infusion system. This transient inhibition of CCL5 activity interrupted hypothalamic insulin signaling, leading to glucose intolerance and peripheral systemic insulin sensitivity.

Hipotalamik insülin sinyal fare beyin içine sürekli uyuşturucu infüzyon sistemi ile periferik glikoz intoleransı için eğitim

Insulin regulates systematic metabolism in the hypothalamus and the peripheral insulin response. An inflammatory reaction in peripheral adipose tissues ...

Study of In Vivo Glucose Metabolism in High-fat Diet-fed Mice Using Oral Glucose Tolerance Test (OGTT) and Insulin Tolerance Test (ITT)

Obesity represents the most important single risk factor in the pathogenesis of type 2 diabetes, a disease which is characterized by a resistance to insulin-stimulated glucose uptake and a gross decompensation of systemic glucose metabolism. Despite considerable progress in the understanding of glucose metabolism, the molecular mechanisms of its regulation in health and disease remain under-investigated, while novel approaches to prevent and treat diabetes are urgently needed. Diet derived glucose stimulates the pancreatic secretion of insulin, which serves as the principal regulator of cellular anabolic processes during the fed-state and thus balances blood glucose levels to maintain systemic energy status. Chronic overfeeding triggers meta-inflammation, which leads to alterations in peripheral insulin receptor-associated signaling and thus reduces the sensitivity to insulin-mediated glucose disposal. These events ultimately result in elevated fasting glucose and insulin levels as well as a reduction in glucose tolerance, which in turn serve as important indicators of insulin resistance. Here, we present a protocol for the generation and metabolic characterization of high-fat diet (HFD)-fed mice as a frequently used model of diet-induced insulin resistance. We illustrate in detail the oral glucose tolerance test (OGTT), which monitors the peripheral disposal of an orally administered glucose load and insulin secretion over time. Additionally, we present a protocol for the insulin tolerance test (ITT) to monitor whole-body insulin action. Together, these methods and their downstream applications represent powerful tools to characterize the general metabolic phenotype of mice as well as to specifically assess alterations in glucose metabolism. They may be especially useful in the broad research field of insulin resistance, diabetes and obesity to provide a better understanding of pathogenesis as well as to test the effects of therapeutic interventions.

Study of In Vivo Glucose Metabolism in High-fat Diet-fed Mice Using Oral Glucose Tolerance Test OGTT and Insulin Tolerance Test ITT

The current article describes the generation and metabolic characterization of high-fat diet-fed mice as a model of diet-induced insulin resistance and obesity. It further features detailed protocols to perform the oral glucose tolerance test and the insulin tolerance test, monitoring whole-body alterations of glucose metabolism in vivo.

Vivo glikoz metabolizması farelerde Oral glukoz tolerans testi (OGTT) ve insülin tolerans testi (ITT) kullanarak yüksek yağlı diyet beslenen incelenmesi

Obesity represents the most important single risk factor in the pathogenesis of type 2 diabetes, a disease which is characterized by a resistance to ...

Induction of Nephrotic Syndrome in Mice by Retrobulbar Injection of Doxorubicin and Prevention of Volume Retention by Sustained Release Aprotinin

Nephrotic syndrome is the most extreme manifestation of proteinuric kidney disease and characterized by heavy proteinuria, hypoalbuminemia, and edema due to sodium retention and hyperlipidemia. To study the pathophysiology of this syndrome, rodent models have been developed based on the injection of toxic substances such as doxorubicin causing podocyte damage. In mice, only few strains are susceptible to this model. In wildtype 129S1/SvImJ mice, the administration of doxorubicin by rapid intravenous injection to the retrobulbar sinus induces experimental nephrotic syndrome that features all the symptoms of human disease including sodium retention and edema. After the onset of proteinuria, mice exhibit increased urinary serine protease activity that leads to the activation of the epithelial sodium channel (ENaC) and sodium retention. Pharmacological inhibition of urinary serine proteases by the treatment with sustained release aprotinin abrogates ENaC activation and prevents sodium retention. This model is ideal to study the pathophysiology of proteasuria, i.e., the excretion of active serine proteases that cause ENaC activation by the proteolysis of its &#947;-subunit. This can be regarded as the primary mechanism of ENaC activation and sodium retention in proteinuric kidney disease.

Here, we describe the induction of experimental nephrotic syndrome in 129S1/SvImJ mice by rapid retrobulbar injection of doxorubicin. We also treat nephrotic mice with sustained release pellets containing aprotinin to inhibit urinary serine protease activity and prevent sodium retention.

Doksorubisin Retrobulbar enjeksiyon ve Önleme Birimi kalıcılık tarafından sürekli yayın Aprotinin farelerde Nefrotik sendromda indüksiyon

Nephrotic syndrome is the most extreme manifestation of proteinuric kidney disease and characterized by heavy proteinuria, hypoalbuminemia, and edema due ...

Mechanisms Underlying Gut Hormone Secretion Using the Isolated Perfused Rat Small Intestine

The gut is the largest endocrine organ of the body, producing more than 15 different peptide hormones that regulate appetite and food intake, digestion, nutrient absorption and distribution, and post-prandial glucose excursions. Understanding the molecular mechanisms that regulate gut hormone secretion is fundamental for understanding and translating gut hormone physiology. Traditionally, the mechanisms underlying gut hormone secretion are either studied in vivo (in experimental animals or humans) or using gut hormone-secreting primary mucosal cell cultures or cell lines. Here, we introduce an isolated perfused rat small intestine as an alternative method for studying gut hormone secretion. The virtues of this model are that it relies on the intact gut, meaning that it recapitulates most of the physiologically important parameters responsible for the secretion in in vivo studies, including mucosal polarization, paracrine relationships and routes of perfusion/stimulus exposure. In addition, and unlike in vivo studies, the isolated perfused rat small intestine allows for almost complete experimental control and direct assessment of secretion. In contrast to in vitro studies, it is possible to study both the magnitude and the dynamics of secretion and to address important questions, such as what stimuli cause secretion of different gut hormones, from which side of the gut (luminal or vascular) is secretion stimulated, and to analyze in detail molecular sensors underlying the secretory response. In addition, the preparation is a powerful model for the study of intestinal absorption and details regarding the dynamics of intestinal absorption including the responsible transporters.

Here, we present a powerful and physiological model to study the molecular mechanisms underlying gut hormone secretion and intestinal absorption &#8212; the isolated perfused rat small intestine.

Gut hormonu salgılanması izole kullanarak temel mekanizmaları fare bağırsağı periosteum

The gut is the largest endocrine organ of the body, producing more than 15 different peptide hormones that regulate appetite and food intake, digestion, ...

Incorporation of a Survivable Liver Biopsy Procedure in Mice to Assess Non-alcoholic Steatohepatitis (NASH) Resolution

Clinical trials assessing therapies for the treatment of non-alcoholic steatohepatitis (NASH) involve a baseline and end of study liver biopsy, and assessment of improvement in disease endpoints, often reflected as a percent of each treatment arm that improved, worsened or remained unchanged. Traditional preclinical rodent studies for putative NASH therapies are often limited by not knowing the level of liver disease/NASH present at the start of therapeutic intervention, instead of randomizing treatment groups on easily measurable endpoints such as body weight, metabolic status or similar. Here, we describe a liver biopsy technique in a diet-induced NASH mouse model, for the assessment of baseline liver disease in order to exclude mice that do not exhibit fibrosis and to equally distribute animals with similar fibrosis between treatment groups. These levels can then be compared to the terminal, post-intervention levels for a truer understanding of in vivo pharmacological effects and thus more accurately reflect clinical trial design strategies. The mouse is properly anesthetized and prepared for the surgery using sterile conditions. A small incision is made in the upper abdomen and the left lateral lobe of the liver is exposed. A wedge of the liver is surgically removed, and a similar-sized piece of absorbable gelatin is put in its place to stop any bleeding. The mouse is surgically sutured and stapled closed and will recover back to normal within 1 day. The entire process takes 5-10 min per mouse. Here we exemplify the utility of this procedure by leveraging the pre-study biopsy to assess the impact of the glucagon-like peptide-1 (GLP-1) receptor agonist liraglutide on NASH endpoints in mice.

Incorporation of a Survivable Liver Biopsy Procedure in Mice to Assess Non-alcoholic Steatohepatitis NASH Resolution

In clinical settings, a liver biopsy is used to assess stages of non-alcoholic steatohepatitis (steatosis, inflammation, hepatocyte ballooning, and fibrosis). Here, we illustrate a survivable liver biopsy in mice which can be used for histological analyses to enable assessment of therapeutic agents in a manner more aligned with clinical trials.

-Alkolsüz Steatohepatitis (NASH) çözünürlük değerlendirmek farelerde hayatta kalınabilir karaciğer biyopsisi yordam eklenmesi

Clinical trials assessing therapies for the treatment of non-alcoholic steatohepatitis (NASH) involve a baseline and end of study liver biopsy, and ...