Yazarlar, Ulusal Bilim Vakfı (NSF) mali destek &quot;Sağlık ve Çevre için Orta-Kızılötesi Teknolojileri (MIRTHE)&quot; başlıklı EEC-0540832 hibe kabul

Araçların 1. Hazırlanması <ol><li> Amonyak optik sensör platformu, lazer diyot denetleyici, özel inşa kontrol elektroniği ünitesi (CEU), nefes örnekleyici, hava pompası, ve dizüstü bilgisayar, harici güç kaynağı açın. </li><li> Amonyak sensörü egzoz ve soğutma fanları hem çalışıp çalışmadığını kontrol edin. NOT: Bir sensörün arka bulunan, ikinci kolayca erişilebilir sensör, içinde bulunur. </li><li> Akustik algılama modülü ve iğne vana sıcaklık amonyak sensörü kutusunun tarafında bulunan dijital göstergeyi kontrol ederek 38.0 ° C&#39;de olduğundan emin olun. Sensör stabilize etmek için sıcaklık için güç sağlanan zaman yaklaşık 35 dakika bekleyin. </li><li> Bir simge isim nefes numune masaüstünde bulunan &quot;değişimi t&quot; tıklayarak 55 ° C giriş tüp ve ağızlık sıcaklığı ayarlayın. Daha sonra sıcaklık &quot;Upd tıklayarak takip yukarı veya aşağı oku tıklayarak değiştirilebilir&quot;Temp yedik. Masaüstüne dönmek için &quot;Exit&quot; ı tıklayın. Sistem stabilize etmek için sıcaklık için en az 5 dakika izin verin. 55 ° C&#39;de numune ve boru ısısını amonyak yüzey kaybını en aza indirir. </li><li> Amonyak sensörü kontrol laptop yazılımını açın. Program masaüstünde &quot;NH 3 Nefes Sensör Programı&quot; adlı bir klasör içinde ulaşılabilir. Bu klasör içinde, kullanıcı simgesini &quot;ana labview yazılım&quot; seçmelisiniz. Bu klasör çeşitli uygulamaları içerir, ancak kullanıcı istenen arayüzüne erişmek için &quot;Mainsequence.vi&quot; seçmelisiniz. Ekranın sol üst köşesindeki &quot;Çalıştır&quot; seçeneğini seçin. NOT: Bu kalibrasyon dizisi kilitleme satır başlar. QEPAS monitörün Lazer otomatik hat-kilitleme prosedürü sırasında seçilmiş optimal akım, çalışır. Bu işlem yaklaşık 25 dakika sürer. </li><li> Örnekleyicideki yeni bir oturum oluşturma şekilde adetch konu oturumu bağlı bir flash sürücüye kaydedilir kendi dosyası vardır. NOT: Bu numune masaüstünde bulunan programı &quot;Nefes Sampler&quot; açarak yapılır. Uygun oturumu tanımlamak alanı var. Oturumu sırasında oluşturulan tüm veriler bu tanımlayıcı altında flash sürücüye kaydedilir. Deneyin tarih tipik olarak dosya adının bir parçası olarak kullanılır. Giriş ve ağızlık sıcaklığının nefes numune alma programı girmeden önce ayarlanmalıdır. </li><li> Giriş borusu yeni bir tek kullanımlık ağızlık yerleştirin. Parmaklardan amonyak ile ağızlık kirlenmesini önlemek için tek kullanımlık eldiven giyin. </li></ol> 2.. Nefes Örnek Toplama NOT: İlgili Kurumsal Değerlendirme Kurulu (Etik Kurulu), insan denekler içeren herhangi bir çalışma onaylaması gerekir. Büyük ölçüde nefes amonyak etkileyebilecek pek çok faktör vardır. Bu faktörler doğrudan tarafından nefes amonyak ölçümleri değiştirebilirsistemik amonyak seviyelerini ffecting veya enstrümantasyon akciğerlerden nefes metabolitleri yolculuk etkileyerek. <ol><li> Konular girişte yaklaşık 12 saat boyunca ve test öncesinde egzersiz sabah kaçındı var ki hiçbir yiyecek tüketmiş olan, bir açlık durumda laboratuara gelmesi emin olun. </li><li> Hiçbir madde, veri toplamadan önce en az 1 saat boyunca, ağız içine getirilmiştir emin olun. Denekler test 1 saat önce daha da dişlerin daha büyük fırça emin olun. </li><li> Amonyak sensörü önünde konuyu koltuk. Giriş borusunu tutun, ve onlar amonyak kontaminasyonu önlemek için ağızlığı dokunmayın emin olmak için konuyu söyleyin. </li><li> Örnekleyici arayüzünde &quot;Başlat&quot; düğmesine tıklayın. Konu sürece onlar gibi için ağızlık içine nefes, ya da operatör yeterli olması numuneyi gördüğü kadar var. NOT: Bu en az 10 saniye süren bir tek tam nefes olduğunu. Ağız basınç bir vekil olarak gerçek zamanlı olarak ölçülürakış hızı için. Bir renk kodlu basınç göstergesi özne üretmek yardımcı olur ve 50 ml / sn bir soluma akış hızını temsil suyun 10 cm istenilen soluk basıncını korumak. Bu Fe NO belirlemek için protokol için ATS / ERS tarafından kabul beri bu akış hızı seçildi. Bu soluma akış hızı çocuklar ve yetişkinler tarafından başarılabilir. Aynı şekilde, üç tekrarlanabilir ekshalasyonlar az% 10 farklı olduğu alınmalıdır. </li><li> Nefes örneği tamamlandığında örnekleyici arayüzünde &quot;Dur&quot; tıklayın. </li></ol> 3.. Nefes Numune Ölçümü <ol><li> Bir nefes örneği analiz edildikten sonra, monitör çalışırken laboratuar teknisyeni o nefes profilinin herhangi bir segment analiz yeteneğine sahiptir. Ilgi nefes kısmı faz III bölümdür. Bu karbon dioksit konsantrasyonunda bir &quot;yayla&quot; ile karakterize edilir ve bre geç aşamasında ortada bulunanath. </li><li> Doğru nefes basınç düşümü önce zaman karbondioksit yaylaları başlar ve durdurmak için örnekleyici arayüzünde dikey çizgileri sürükleyerek numunenin faz III bölümünü seçin. Açıklama için Bkz: Şekil 1.. </li><li> Örnekleyici dokunmatik ekran arayüzü üzerinde &quot;Mağaza&quot; tıklayarak flash sürücüye verileri kaydetmek. </li><li> Nefes verileri saklandıktan sonra, kullanıcı yeni bir nefes numunesi başlatmak için &quot;başlangıç&quot; seçebilirsiniz. </li></ol> 4. Nefes Amonyak Mouth durulayın ve pH etkilerini gösteren <ol><li> Temel amonyak seviyesini oluşturmak için 3. nefes Örnek. Nefesler birbirinden en az 5 dakika alınır emin olun. </li><li> 60 saniye için bir su 30 mi kısım ile iyice ağız çalkalayın. </li><li> Durulama (Bölüm 2) 60 saniye içinde bir nefes numunesi toplayın. Zaman içinde amonyak değişikliği gözlemek için bir sonraki saat boyunca nefes örnekleri toplamak. NOT: Samples her dakika kadar sık ​​alınabilir, fakat tipik olarak daha büyük zaman aralıkları kullanılır. </li><li> 60 saniye için bazik bir solüsyon 30 ml alikotu (su içinde sodyum bikarbonat) ile iyice yıkayın ve ağız 4.2.1 tekrarlayın. </li><li> 60 saniye için asidik çözelti bir 30 ml alikotu ile iyice yıkayın ve ağız 4.2.1 tekrarlayın. </li></ol> 5.. Nefes Amonyak Inlet ve Ulaştırma Boru Sıcaklıkların etkilerini gösteren <ol><li> Giriş sıcaklığı ile 15 dakikalık bir süre boyunca Örnek, üç nefes vücut sıcaklığı, yaklaşık 30 ° C&#39;nin altında ayarlanmış </li><li> Nefes sampler&#39;ın dokunmatik ekran arayüzü üzerinde bir masaüstü simgesini kullanarak sırasıyla 55 ° C giriş ve taşıma boru sıcaklıklarını artırın. Sistem en az 5 dk kararlı duruma ulaşmak için izin verin. </li><li> 5 dk arayla ısıtılmış girişine Numune 3 nefesler. </li></ol>

<ol>
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H. <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+ammonia+in+biological+fluids.">Determination of ammonia in biological fluids.</a> Annals of clinical biochemistry. 31 (Pt 6), 529-543 (1994).</li><li>Lewicki, 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=Real+time+ammonia+detection+in+exhaled+human+breath+with+a+quantum+cascade+laser+based+sensor.">Real time ammonia detection in exhaled human breath with a quantum cascade laser based sensor.</a> 2009 Conference on Lasers and ElectroOptics and 2009 Conference on Quantum electronics and Laser Science Conference. 1, (2009).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=American+Thoracic+Society.+European+Respiratory+Society.+Recommendations+for+Standardized+Procedures+for+the+Online+and+Offline+Measurement+of+Exhaled+Lower+Respiratory+Nitric+Oxide+and+Nasal+Nitric+Oxide.">American Thoracic Society. European Respiratory Society. Recommendations for Standardized Procedures for the Online and Offline Measurement of Exhaled Lower Respiratory Nitric Oxide and Nasal Nitric Oxide.</a> American journal of respiratory and critical care medicine. 171 (8), 912-930 (2005).</li><li>Solga, S. F., 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=Factors+influencing+breath+ammonia+determination.">Factors influencing breath ammonia determination.</a> Journal of breath research. 7 (3), (2013).</li><li>Schmidt, F. M., 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=Ammonia+in+breath+and+emitted+from+skin.">Ammonia in breath and emitted from skin.</a> Journal of breath research. 7 (1), (2013).</li><li>Span&#283;l, P., Dryahina, K., Smith, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+quantitative+study+of+the+influence+of+inhaled+compounds+on+their+concentrations+in+exhaled+breath.">A quantitative study of the influence of inhaled compounds on their concentrations in exhaled breath.</a> Journal of breath research. 7 (1), (2013).</li><li>Boots, A. W., van Berkel, J. J. B. N., Dallinga, J. W., Smolinska, A., Wouters, E. F., van Schooten, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+versatile+use+of+exhaled+volatile+organic+compounds+in+human+health+and+disease.">The versatile use of exhaled volatile organic compounds in human health and disease.</a> Journal of breath research. 6 (2), (2012).</li><li>Hibbard, T., Killard, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breath+ammonia+levels+in+a+normal+human+population+study+as+determined+by+photoacoustic+laser+spectroscopy.">Breath ammonia levels in a normal human population study as determined by photoacoustic laser spectroscopy.</a> Journal of breath research. 5 (3), (2011).</li><li>Wang, T., Pysanenko, A., Dryahina, K., Span&#283;l, P., Smith, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+breath,+exhaled+via+the+mouth+and+nose,+and+the+air+in+the+oral+cavity.">Analysis of breath, exhaled via the mouth and nose, and the air in the oral cavity.</a> Journal of breath research. 2 (3), (2008).</li><li>Amann, A., Smith, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Volatile Biomarkers. 1st Edition. , (2013).</li></ol>

Dr. Risby serves as a consultant for Loccioni Humancare the manufacturer of the breath sampler.

Gerçek zamanlı izleme metabolitleri tespit edebilen bir non-invazif yararları açıktır. Ancak, nefes araştırma alanında bu potansiyeli yerine getirmek için mücadele etti. Nefes ölçüm birçok karıştırıcı faktörler savunmasız dinamik bir süreçtir. Bizim yaklaşım önemli artıları vardır: özellikle, nefes örnekleyiciye birleştiğinde Rice QEPAS dayalı amonyak monitörün hassasiyet ve hız doğru ölçümü germane nefes toplama faktörleri değerlendirmek ve tanımlamak için bize sağlamış...

Amonyak protein metabolizması 1 bir yerde yan ürünü. Amonyak ölçümü nedenle klinisyenler, çeşitli hastalık ve sağlık durumları 2 değerlendirmeye yardımcı olabilir. Ancak, amonyak çok reaktif olduğundan, kan ya da nefes yoluyla doğru ölçülmesi zordur. Yaygın olarak kullanılan rağmen, kan deneyleri, 3 doğruluğu ile ilgili temel sorunlardan dahil olmak üzere sayısız dezavantajları vardır. Ancak kan tahlilleri ile büyük sorun onlar sadece hiç episodically toplanan gerçektir. Amonyak fizyolojisi, çok kan glukoz ve diğer birçok metabolik süreçler gibi, akışkan ve sürekli 4 değişiyor, çünkü bu önemlidir. Buna karşılık, nefes deneyler böylece kolayca tekrarlanan önlemleri sağlayan, tamamen non-invaziv ve hızlı. Benzersiz bir şekilde ciddi bir karşılanmamış ihtiyaca hitap edebilir çünkü Böylece, nefes amonyak ölçüm çekici. Nefes toplama, ancak, benzersiz endişeleri sunuyor. Flebotomi doğal JEO taşır oysaçeşitli öngörülemeyen şekilde hata PARDY (örneğin, turnike zaman, ter kirlenme, kan hücresi hemoliz, laboratuar ölçümü gecikme, vb 5), nefes ölçüm araştırmacılar yeni zorluklar farklı bir grup ile uğraşmak gerekir: değişkenliği nefes, kirlenmeyi ve ağız mukozası ile ya bakteriyel amonyak, ortam havası ve cihaz nem ve sıcaklığın etkisi, vs 6. Gerçekten de, bu bilinmeyen biyoloji keşfetmek için deneysel yöntemler kullanılarak insanlarda deneysel ekipman bağlayan görevi hafife akılsızca. Nedeniyle bu engellere kısmen, nefes amonyak henüz potansiyelini bir araya geldi değil. Bu yazıda, hızlı ve doğru sonuçlar için bizim nefes amonyak ölçüm protokolü sunuyoruz. Monitör, arayüz sampler ve insan etkilerine dikkat: Bizim protokol üç alanda gücü vardır. Monitör, daha önce açıklandığı gibi 7 Rice Üniversitesi&#39;nde meslektaşları tarafından inşa edilmiştir. Mea temelisurement bir kuvars gelişmiş foto akustik spektroskopi bir akustik dönüştürücü olarak bir piezoelektrik diyapazonum istihdam (QEPAS) tekniğidir. Akustik dalgalar hedef izleme gaz türleri tarafından modüle Lazer radyasyonu emme tarafından üretilen zaman foto akustik etki meydana gelir. Izleme gaz ile modüle edilmiş frekans rezonant akustik bir akustik hücre kullanılarak tespit edilir. Amonyak için bir absorpsiyon dalga boyu o nefeste türlerin müdahale spektral müdahalelerden arınmış seçildi. İnsan Solunan nefes ölçüm amaçları için, monitörün ana özellikleri, geniş ölçüm aralığı (50 parça milyar başına ~, en az 5.000 ppb ppb) ve hız (1 sn ölçümleri) içerir. Monitörün hızı nefes döngüsü boyunca zaman çözünürlüğü sağlar. Monitör, özel olarak tasarlanmış bir nefes örnekleyici kuple edilir. Örnekleyici bir basınç sensörü ve kapnograf oluşur. Bu görüntüler ve gerçek zamanlı arşivlerağız basıncı ve karbon dioksit gibi, sensör tarafından belirlenen amonyak konsantrasyonlarının ölçümleri. Bu örneği, bu nedenle, nefes toplanır gibi nefes çaba kalitesini değerlendirmek için teknisyen sağlar. Bu Amerikan Toraks Derneği / Avrupa Solunum Derneği Görev Gücü (ATS / ERS) 8 önerdiği nefes nitrik oksit (NO Fe) analiz için öneriler aşmak için bize sağlar. Her nefes örneği alma için, tek tek yönlü sıralı valf nefes numune ağız noktası kullanıldı. Çünkü örnekleyici tarafından sağlanan monitör ve kalite kontrolleri hızı, dikkatle 9 insan etkilerini değerlendirmeyi başardık. Nefes talimatı deneklerin çoğu, örneğin, başlangıçta hyperventilate. Böyle bir oral pH ve ağız gibi diğer önemli etkiler, örnekleyici, monitör ve tüm ilişkili boru ve nefes modu sıcaklıkları, sonra çalışıldı çalkalanması ve temel fo vardırAşağıda r örnek deneyleri. Son olarak ve belki de en önemlisi, birden çok deneyimli gruplar tamamen farklı sensörler ve ölçüm yöntemleri kullanılarak nefes amonyak ölçüm olduğunu vurgulamak gerekir. Bu önemli avantaj ve geçerliliği olabilir. Tam bir karşılaştırma mevcut çalışma 10,11,12 kapsamı dışındadır.

<table border="0" cellpadding="0" cellspacing="0" width="666">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Rice Ammonia Monitor System</td>
			<td style="width:171px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td style="width:167px;">Not available for commercial purchase</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Loccioni Breath Sampler</td>
			<td style="width:171px;">Loccioni Humancare</td>
			<td style="width:119px;">N/A</td>
			<td>Single breath version</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Disposable Mouth Piece</td>
			<td style="width:171px;">WestPrime Healthcare</td>
			<td style="width:119px;">G011-200</td>
			<td>Manufacturer is AlcoQuant</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Laptop</td>
			<td style="width:171px;">Lenovo</td>
			<td style="width:119px;">N/A</td>
			<td>Old model no longer sold by manufacturer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Acid Rinse</td>
			<td style="width:171px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>Household acidic drink (coffee, soft drink, citrus juices, etc)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Base Rinse</td>
			<td style="width:171px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>Water mixed with a nonexact amount of sodium bicarbonate (Arm &#38; Hammer Baking Soda)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Neutral Rinse</td>
			<td style="width:171px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>Water</td>
		</tr>
	</tbody>
</table>

fast and accurate exhaled breath ammonia measurement

This exhaled breath ammonia method uses a fast and highly sensitive spectroscopic method known as quartz enhanced photoacoustic spectroscopy (QEPAS) that uses a quantum cascade based laser. The monitor is coupled to a sampler that measures mouth pressure and carbon dioxide. The system is temperature controlled and specifically designed to address the reactivity of this compound. The sampler provides immediate feedback to the subject and the technician on the quality of the breath effort. Together with the quick response time of the monitor, this system is capable of accurately measuring exhaled breath ammonia representative of deep lung systemic levels. 
Because the system is easy to use and produces real time results, it has enabled experiments to identify factors that influence measurements. For example, mouth rinse and oral pH reproducibly and significantly affect results and therefore must be controlled. Temperature and mode of breathing are other examples. As our understanding of these factors evolves, error is reduced, and clinical studies become more meaningful. This system is very reliable and individual measurements are inexpensive. 
The sampler is relatively inexpensive and quite portable, but the monitor is neither. This limits options for some clinical studies and provides rational for future innovations.

Denekler bazal nefes amonyak seviyeleri geniş bir yelpazede üretmek beklenebilir. Sağlıklı bireyler 100-1.000 ppb bir nefes amonyak ölçümü ile gün başlayabilir. Herhangi bir sıvı ile ağız dışarı durulama hemen saptanabilir nefes amonyak miktarını değiştirir. Nötr ve asidik sıvılar genellikle yarıdan fazla gözlemlenebilir amonyak miktarı kesti. Durulama etkileri kapalı giyer gibi bu seviyeleri daha sonra başlangıca dönün. Suyun etkisi bir asit 2 saat boyunca fazla bir minimum bulgulanabil...

Watch this Scientific Journal Video about Fast and Accurate Exhaled Breath Ammonia Measurement at JoVE.com

Fast and Accurate Exhaled Breath Ammonia Measurement

Ammonia is an important physiologic metabolite relevant to various disease and wellness states. It is also a difficult molecule to measure in breath, which demands particular precautions be taken to obtain accurate results. Not all factors influencing ammonia are known, but progress can be difficult without accounting for these factors.

Hızlı ve Doğru Üfürükle Amonyak Ölçüm

This exhaled breath ammonia method uses a fast and highly sensitive spectroscopic method known as quartz enhanced photoacoustic spectroscopy (QEPAS) that ...

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13.1K Görüntüleme.  St. Luke's University Hospital. Ammonia is an important physiologic metabolite relevant to various disease and wellness states. It is also a difficult molecule to measure in breath, which demands particular precautions be taken to obtain accurate results. Not all factors influencing ammonia are known, but progress can be difficult without accounting for these factors.

Video: Fast and Accurate Exhaled Breath Ammonia Measurement

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&beta; using a Whole Blood Stimulation Assay

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues as well as altered cytokine homeostasis. The innate immune system plays a crucial role in recognizing pathogens and other endogenous danger stimuli. One of the major cytokines released by innate immune cells is Interleukin (IL)-1. Therefore, we utilize a whole blood stimulation assay in order to measure the secretion of inflammatory cytokines and specifically of the pro-inflammatory cytokine IL-1&beta; 1, 2, 3.
Patients with genetic dysfunctions of the innate immune system causing autoinflammatory syndromes show an exaggerated release of mature IL-1&beta; upon stimulation with LPS alone. In order to evaluate the innate immune component of patients who present with inflammatory-associated pathologies, we use a specific immunoassay to detect cellular immune responses to pathogen-associated molecular patterns (PAMPs), such as the gram-negative bacterial endotoxin, lipopolysaccharide (LPS). These PAMPs are recognized by pathogen recognition receptors (PRRs), which are found on the cells of the innate immune system 4, 5, 6, 7. A primary signal, LPS, in conjunction with a secondary signal, ATP, is necessary for the activation of the inflammasome, a multiprotein complex that processes pro-IL-1&beta; to its mature, bioactive form 4, 5, 6, 8, 9, 10.
The whole blood assay requires minimal sample manipulation to assess cytokine production when compared to other methods that require labor intensive isolation and culturing of specific cell populations. This method differs from other whole blood stimulation assays; rather than diluting samples with a ratio of RPMI media, we perform a white blood cell count directly from diluted whole blood and therefore, stimulate a known number of white blood cells in culture 2. The results of this particular whole blood assay demonstrate a novel technique useful in elucidating patient cohorts presenting with autoinflammatory pathophysiologies.

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&#946; using a Whole Blood Stimulation Assay

We describe a simple immunoassay to measure the production of pro-inflammatory cytokines, such as IL-1 beta production, in patients presenting with autoinflammatory phenotypes. By activating cells in whole blood cultures with pathogen-associated molecular patterns, specifically with lipopolysaccharide, cytokine secretion can be conveniently evaluated in whole blood supernatants.

Tam Kan Stimülasyon Testi kullanarak Pro-inflamatuar sitokin IL-1β Doğru ve Basit Ölçme

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues ...

Tıp

Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye

The presence of placental vascular anastomoses is a conditio sine qua non for the development of twin-to-twin transfusion syndrome (TTTS) and twin anemia polycythemia sequence (TAPS)1,2. Injection studies of twin placentas have shown that such anastomoses are almost invariably present in monochorionic twins and extremely rare in dichorionic twins1. Three types of anastomoses have been documented: from artery to artery, from vein to vein and from artery to vein. Arterio-venous (AV) anastomoses are unidirectional and are referred to as "deep" anastomoses since they proceed through a shared placental cotyledon, whereas arterio-arterial (AA) and veno-venous (VV) anastomoses are bi-directional and are referred to as "superficial" since they lie on the chorionic plate. Both TTTS and TAPS are caused by net imbalance of blood flow between the twins due to AV anastomoses. Blood from one twin (the donor) is pumped through an artery into the shared placental cotyledon and then drained through a vein into the circulation of the other twin (the recipient). Unless blood is pumped back from the recipient to the donor through oppositely directed deep AV anastomoses or through superficial anastomoses, an imbalance of blood volumes occurs, gradually leading to the development of TTTS or TAPS. The presence of an AA anastomosis has been shown to protect against the development of TTTS and TAPS by compensating for the circulatory imbalance caused by the uni-directional AV anastomoses1,2. 
Injection of monochorionic placentas soon after birth is a useful mean to understand the etiology of various (hematological) complications in monochorionic twins and is a required test to reach the diagnosis of TAPS2. In addition, injection of TTTS placentas treated with fetoscopic laser surgery allows identification of possible residual anastomoses3-5. This additional information is of paramount importance for all perinatologists involved in the management and care of monochorionic twins with TTTS or TAPS. Several placental injection techniques are currently being used. We provide a simple protocol to accurately evaluate the presence of (residual) vascular anastomoses using colored dye injection.

Twin-to-twin transfusion syndrome and twin anemia polycythemia sequence are two potentially devastating problems in perinatal medicine. Both disorders occur only in monochorionic twins and result from unbalanced blood flow through placental vascular anastomoses. We provide a simple protocol to accurately evaluate the presence of vascular anastomoses using colored dye injection of placental vessels after birth.

Renkli Boya kullanarak Monokoryonik Plasenta vasküler anastomozlar doğru ve basit Değerlendirme

The presence of placental vascular anastomoses is a conditio sine qua non for the development of twin-to-twin transfusion syndrome (TTTS) and twin anemia ...

Remote Magnetic Navigation for Accurate, Real-time Catheter Positioning and Ablation in Cardiac Electrophysiology Procedures

New remote navigation systems have been developed to improve current limitations of conventional manually guided catheter ablation in complex cardiac substrates such as left atrial flutter. This protocol describes all the clinical and invasive interventional steps performed during a human electrophysiological study and ablation to assess the accuracy, safety and real-time navigation of the Catheter Guidance, Control and Imaging (CGCI) system. Patients who underwent ablation of a right or left atrium flutter substrate were included. Specifically, data from three left atrial flutter and two counterclockwise right atrial flutter procedures are shown in this report. One representative left atrial flutter procedure is shown in the movie. This system is based on eight coil-core electromagnets, which generate a dynamic magnetic field focused on the heart. Remote navigation by rapid changes (msec) in the magnetic field magnitude and a very flexible magnetized catheter allow real-time closed-loop integration and accurate, stable positioning and ablation of the arrhythmogenic substrate.

This report provides a detailed description of a new remote navigation system based on magnetic driven forces, which has been recently introduced as a new robotic tool for human cardiac electrophysiology procedures.

Kardiyak Elektrofizyoloji İşlemleri Doğru, gerçek zamanlı Kateter Konumlandırma ve Ablasyon için uzaktan Manyetik Navigasyon

New remote navigation systems have been developed to improve current limitations of conventional manually guided catheter ablation in complex cardiac ...

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

Ambliyopi içinde Bastırma Ölçme ve Tedavisi

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used techniques require killing of the animals and postmortem recovery of the meal1,2. This approach prevents longitudinal studies to determine changes in gastric emptying with age and progression of disease. The commonly used [13C]-octanoic acid breath test for humans3 has been modified for use in mice4-6 and rats7 and we previously showed that this test is reliable and responsive to changes in gastric emptying in response to drugs and during diabetic disease progression8. In this video presentation the principle and practical implementation of this modified test is explained. As in the previous study, NOD LtJ mice are used, a model of type 1 diabetes9. A proportion of these mice develop the symptoms of gastroparesis, a complication of diabetes characterized by delayed gastric emptying without mechanical obstruction of the stomach10.
This paper demonstrates how to train the mice for testing, how to prepare the test meal and obtain 4 hr gastric emptying data and how to analyze the obtained data. The carbon isotope analyzer used in the present study is suitable for the automatic sampling of the air samples from up to 12 mice at the same time. This technique allows the longitudinal follow-up of gastric emptying from larger groups of mice with diabetes or other long-standing diseases.

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Determination of gastric emptying with a non-invasive [13C]-octanoic acid breath test for tracking gastroparesis in female NOD LtJ mice.

Bir Kullanarak Non-obez Diyabetik Farelerde Mide boşaltılması Değerlendirilmesi [<sup&gt; 13</sup&gt; C]-oktanoik asit Nefes Testi

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used ...

Lesion Explorer: A Video-guided, Standardized Protocol for Accurate and Reliable MRI-derived Volumetrics in Alzheimer's Disease and Normal Elderly

Obtaining in vivo human brain tissue volumetrics from MRI is often complicated by various technical and biological issues. These challenges are exacerbated when significant brain atrophy and age-related white matter changes (e.g. Leukoaraiosis) are present. Lesion Explorer (LE) is an accurate and reliable neuroimaging pipeline specifically developed to address such issues commonly observed on MRI of Alzheimer's disease and normal elderly. The pipeline is a complex set of semi-automatic procedures which has been previously validated in a series of internal and external reliability tests1,2. However, LE's accuracy and reliability is highly dependent on properly trained manual operators to execute commands, identify distinct anatomical landmarks, and manually edit/verify various computer-generated segmentation outputs.
LE can be divided into 3 main components, each requiring a set of commands and manual operations: 1) Brain-Sizer, 2) SABRE, and 3) Lesion-Seg. Brain-Sizer's manual operations involve editing of the automatic skull-stripped total intracranial vault (TIV) extraction mask, designation of ventricular cerebrospinal fluid (vCSF), and removal of subtentorial structures. The SABRE component requires checking of image alignment along the anterior and posterior commissure (ACPC) plane, and identification of several anatomical landmarks required for regional parcellation. Finally, the Lesion-Seg component involves manual checking of the automatic lesion segmentation of subcortical hyperintensities (SH) for false positive errors.
While on-site training of the LE pipeline is preferable, readily available visual teaching tools with interactive training images are a viable alternative. Developed to ensure a high degree of accuracy and reliability, the following is a step-by-step, video-guided, standardized protocol for LE's manual procedures.

Lesion Explorer (LE) is a semi-automatic, image-processing pipeline developed to obtain regional brain tissue and subcortical hyperintensity lesion volumetrics from structural MRI of Alzheimer's disease and normal elderly. To ensure a high level of accuracy and reliability, the following is a video-guided, standardized protocol for LE's manual procedures.

Lezyon Explorer: Doğru ve Güvenilir MR türetilmiş Alzheimer Hastalığı Volumetrics ve Normal Yaşlı Bir Video-güdümlü, Standart Protokol

Obtaining in vivo human brain tissue volumetrics from MRI is often complicated by various technical and biological issues. These challenges are ...

Voluntary Breath-hold Technique for Reducing Heart Dose in Left Breast Radiotherapy

Breath-holding techniques reduce the amount of radiation received by cardiac structures during tangential-field left breast radiotherapy. With these techniques, patients hold their breath while radiotherapy is delivered, pushing the heart down and away from the radiotherapy field. Despite clear dosimetric benefits, these techniques are not yet in widespread use. One reason for this is that commercially available solutions require specialist equipment, necessitating not only significant capital investment, but often also incurring ongoing costs such as a need for daily disposable mouthpieces. The voluntary breath-hold technique described here does not require any additional specialist equipment. All breath-holding techniques require a surrogate to monitor breath-hold consistency and whether breath-hold is maintained. Voluntary breath-hold uses the distance moved by the anterior and lateral reference marks (tattoos) away from the treatment room lasers in breath-hold to monitor consistency at CT-planning and treatment setup. Light fields are then used to monitor breath-hold consistency prior to and during radiotherapy delivery.

The current priority in breast cancer radiotherapy is to reduce cardiac doses without compromising target tissue coverage. The voluntary breath-hold technique described here is a simple, inexpensive solution to this problem and capable of being instituted widely without the need for specialized equipment.

Sol Meme Radyoterapi Kalp Doz azaltılması için gönüllü Nefes tutma Tekniği

Breath-holding techniques reduce the amount of radiation received by cardiac structures during tangential-field left breast radiotherapy. With these ...

Creating Anatomically Accurate and Reproducible Intracranial Xenografts of Human Brain Tumors

Orthotopic tumor models are currently the best way to study the characteristics of a tumor type, with and without intervention, in the context of a live animal &#8211;&#160;particularly in sites with unique physiological and architectural qualities such as the brain. In vitro and ectopic models cannot account for features such as vasculature, blood brain barrier, metabolism, drug delivery and toxicity, and a host of other relevant factors. Orthotopic models have their limitations too, but with proper technique tumor cells of interest can be accurately engrafted into tissue that most closely mimics conditions in the human brain. By employing methods that deliver precisely measured volumes to accurately defined locations at a consistent rate and pressure, mouse models of human brain tumors with predictable growth rates can be reproducibly created and are suitable for reliable analysis of various interventions. The protocol described here focuses on the technical details of designing and preparing for an intracranial injection, performing the surgery, and ensuring successful and reproducible tumor growth and provides starting points for a variety of conditions that can be customized for a range of different brain tumor models.

The brain is a unique site with qualities that are not well represented by in vitro or ectopic analyses. Orthotopic mouse models with reproducible location and growth characteristics can be reliably created with intracranial injections using a stereotaxic fixation instrument and a low pressure syringe pump.

Oluşturma Anatomik Doğru ve İnsan Beyin Tümörleri Reproducible İntrakraniyal Ksenograftlar

Orthotopic tumor models are currently the best way to study the characteristics of a tumor type, with and without intervention, in the context of a live ...

3D Ultrasound Imaging: Fast and Cost-effective Morphometry of Musculoskeletal Tissue

The developmental goal of 3D ultrasound imaging (3DUS) is to engineer a modality to perform 3D morphological ultrasound analysis of human muscles. 3DUS images are constructed from calibrated freehand 2D B-mode ultrasound images, which are positioned into a voxel array. Ultrasound (US) imaging allows quantification of muscle size, fascicle length, and angle of pennation. These morphological variables are important determinants of muscle force and length range of force exertion. The presented protocol describes an approach to determine volume and fascicle length of m.&#160;vastus lateralis and m.&#160;gastrocnemius medialis. 3DUS facilitates standardization using 3D anatomical references. This approach provides a fast and cost-effective approach for quantifying 3D morphology in skeletal muscles. In healthcare and sports, information on the morphometry of muscles is very valuable in diagnostics and/or follow-up evaluations after treatment or training.

3D ultrasound imaging (3DUS) allows fast and cost-effective morphometry of musculoskeletal tissues. We present a protocol to measure muscle volume and fascicle length using 3DUS.

3D ultrason görüntüleme: hızlı ve uygun maliyetli morfometri kas-iskelet dokusunun

The developmental goal of 3D ultrasound imaging (3DUS) is to engineer a modality to perform 3D morphological ultrasound analysis of human muscles. 3DUS ...

Fast Inspection of Quality of Indigo Naturalis by Multiple Light Scattering

Quality control of Chinese herbal medicine is a crucial component of Chinese herbal medicine research and development. Faced with the challenges of modernization and internationalization of Chinese herbal medicine, it is urgent to establish thorough and effective procedures for quality identification of Chinese herbal medicine, and there is an urgent need for new analytical and testing techniques that are efficient, accurate, and environmentally friendly. 
Multiple light scattering is a cutting-edge and analytical method that can accurately and rapidly assess the quality of Chinese herbal medicine without altering the nature or state of the sample or using organic reagents. Indigo Naturalis is considered a good remedy for pediatric hyperthermia, psoriasis, leukemia, and ulcerative colitis. In this study, the process of addition of Indigo Naturalis powder in water was recorded precisely using a multiple light scattering instrument. 
The qualitative and quantitative measurements of the instrument can be used to accurately capture the overall trajectory and sinking behavior of Indigo Naturalis powder into water and to establish a rapid evaluation method for the quality of Indigo Naturalis with the transmission and backscattering spectrograms of the sample as qualitative indicators and stability index as a quantitative indicator. The analytical technique based on multiple light scattering provides a fast, accurate, green, and environmentally friendly method for the quality evaluation of Indigo Naturalis and supports the development and transformation of high-quality Indigo Naturalis.

Here, we present a quality evaluation method based on a multiple light scattering technique to evaluate the quality of Indigo Naturalis.

Indigo Naturalis'in kalitesinin çoklu ışık saçılımı ile hızlı bir şekilde incelenmesi

Quality control of Chinese herbal medicine is a crucial component of Chinese herbal medicine research and development. Faced with the challenges of ...