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1. Vorbereitung der Plazenta bei der Geburt <ol><li> Beschriften Sie die Nabelschnur der Zwillinge mit einem (für das erste geboren) oder zwei (für das zweite geboren) Klammern. </li><li> Überprüfen Sie die mütterlichen und fetalen Oberfläche der Plazenta auf Vollständigkeit oder Störung. </li><li> Notieren Sie sich folgende Daten: Art des Nabelschnuransatz (zentral, exzentrisch, marginal oder velamentous), die Anzahl der Blutgefäße in der Nabelschnur (in der Regel eine Vene und zwei Arterien, manchmal nur eine Arterie) und Farbe Unterschied zwischen den beiden plazentaren Anteile. Ein Teil der Division Membranen können zur Pathologie geschickt werden, um die Art der Chorionizität bestätigen. </li><li> Die Plazenta kann dann in eine Plastikschüssel gelegt und gekühlt, bis die Abschlussprüfung (am besten innerhalb einer Woche) und Farbstoff Injektion. </li><li> Die Plazenta ist nicht gefroren oder fixiert werden (keine Formalin). </li></ol> 2. Katheterisierung der Nabelgefässe <ol><li> Waschen Sie die Plazenta mit warmem Wasser oder Kochsalzlösung. </li><li> Schneiden Sie die peripheren Membranen, entfernen Sie die inter-twin Trennmembran und ziehen Sie das Amnion (zur besseren Visualisierung der Gefäßanastomosen und bessere Qualität der Plazenta Bilder). </li><li> Transect jeder Nabelschnur bei ca. 5 cm Entfernung von der Nabelschnuransatz. </li><li> Sanft Blutgerinnsel aus der Nabelschnur Schiffe und Plazentagefäße. </li><li> Die Nabelvene ist in der Regel leicht zu erkennen aufgrund seiner größeren Durchmesser, verglichen mit dem kleineren Durchmesser der beiden Nabelarterien. </li><li> Kanülieren die Nabelvene mit einem passenden Katheter. Vermeiden Sie falsche Wege. </li><li> Kanülieren einen Nabelpunkt Arterie mit einem kleineren Katheter. Sie mit einer Pinzette in das Lumen der A. umbilicalis zu erweitern. Vermeiden Sie falsche Wege. Nur einer der 2 Nabelarterien muss katheterisiert da eine Anastomose (von Hyrtl) verbindet die 2 Arterien in der Nähe des Nabelschnuransatz werden. </li><li> Wiederholen Sie die beiden Schritte für die anderen Nabelschnur. </li><li> Die Platzierung des Katheters kann durch vorsichtiges hin und her massiert der Nabelgefässe erleichtert werden. Jede Art von Katheter kann für dieses Verfahren verwendet werden. Wir wählen zu benutzen (und Recycling) die Katheter in unserem Neonatologie Abteilung für Nabelschnur-Katheterisierung bei Neugeborenen eingesetzt. </li><li> Tie ein Stück Klebeband um beide Schnüre zu vermeiden Rückfluss des Farbstoff während Farbstoff Injektion. </li></ol> 3. Injection mit Farbstoff <ol><li> Schließen Sie eine 20 ml Spritze mit Farbstoff auf jedem Katheter gefüllt. </li><li> Jede viskosen Farbstoff kann die Plazenta Angio-Architektur zu visualisieren. Verwenden Sie kontrastreiche Farben, um eine gute Visualisierung der Anastomosen (dunkle Farben für die Arterien, helle Farben für die Venen) zu ermöglichen. </li><li> Vorsichtig injizieren (mit niedrigem Druck) der Farbstoff in die Vene, während ein Assistent sanft drückt den Farbstoff der Farbstoff ermöglichen, alle Plazentagefäße, auch die Kleinsten zu füllen. </li><li> Achten Sie besonders auf die kleinen Gefäße in der Nähe des vaskulären Äquator (der vaskulären Äquator ist der Ort, wo die Anastomosen aus entweder zwei miteinander verbinden). </li><li> Wiederholen Sie die vorherigen Schritte, um Farbstoff in die Arterie injiziert. Zu beachten ist: Arterien schwieriger sein kann zum Einschleusen und erfordern mehr Geduld. </li><li> Wiederholen Sie die beiden Schritte für die anderen Nabelschnur. </li></ol> 4. Auswertung und Dokumentation der Plazenta nach der Farbstoff Injektion <ol><li> Sorgfältig prüfen Gefäßäquator und notieren Sie die Anzahl und Art der Anastomosen. </li><li> Legen Sie ein Maßband auf die Plazenta in den Durchmessern und Plazenta-Aktien an der digitalen Bild zu messen. </li><li> Verwenden Sie eine hochauflösende digitale Kamera und machen Sie Fotos von der injizierten Plazenta. Stellen Sie sicher, dass die Bilder senkrecht zu der Plazenta entnommen. </li></ol> 5. Repräsentative Ergebnisse: Die Plazenta Angio-Architektur in monochorialen Zwillingen variiert je nach der Art der monochorialen Zwillingsschwangerschaft. Injection-Studien haben gezeigt, dass AA, AV-und VV-Anastomosen in jeweils 80%, 95% und 20% der unkomplizierten monochorialen Zwillingsschwangerschaften 1 (Abbildung 1). AA Anastomosen werden als gegen die Entwicklung von TTTS und TAPS 6 zu schützen. Injektionen Studien haben gezeigt, dass AA Anastomosen in nur 20% der TTTS Plazenten und 10% der TAPS Plazenten 1,2,7 (Abbildung 2) auftreten. In TTTS Plazenten mit fetoscopic Laserchirurgie behandelt wird, kann die Narben, die Laserkoagulation der Gefäßanastomosen verursacht auf der Plazenta (Abbildung 3 und 4) zu sehen. TAPS Plazenten sind durch die Anwesenheit von nur ein paar winzige AV Anastomosen 2 (Abbildung 5) aus. Die Plazenta Anteil der TAPS Empfänger ist oft plethoric, während die Plazenta Anteil der Spender ist blass (Abbildung 6). In monochorialen Zwillingsschwangerschaften mit einem Geburtsgewicht Diskordanz, das Wachstum Fötus beschränkthat oft einen velamentous Nabelschnuransatz und eine viel kleinere Plazenta Aktie (Abbildung 7). AA Anastomosen sind häufig in monochorialen Plazenten Geschenk von Zwillingen mit einem Geburtsgewicht Diskordanz 8. Monochoriale-monoamniotische Plazenten haben eine charakteristische Angio-Architektur mit einem kurzen Abstand zwischen den beiden Kabel Insertionen (Abb. 8). Die Inzidenz von AA Anastomosen in monoamniotische-monochorialen Plazenten ist nahezu 100% und verhindert die Entwicklung von TTTS in monoamniotische Zwillinge 9. Die Zahlen in diesem Artikel zeigen die charakteristischen Befunde der monochorialen Plazenten in unserem Zentrum mit Farbstoff injiziert. Wir routinemäßig in dunkleren Farben (blau oder grün) für Arterien und helleren Farben (gelb, pink oder orange) für Venen. <img alt="Abbildung 1" src="/files/ftp_upload/3208/3208fig1.jpg" /> Abbildung 1. Monochoriale Plazenta von einem normalen, unkomplizierten monochorialen Zwillingsschwangerschaft zeigt mehrere AV Anastomosen von grün bis rosa Arterien Venen (weiße Sterne), mehrere VA Anastomosen von blau nach gelb Arterien Venen (grüne Sterne) und 1 große AA Anastomose (gekennzeichnet durch die Mischen der blauen und grünen Farbstoff, blauer Stern). <img alt="Abbildung 2" src="/files/ftp_upload/3208/3208fig2.jpg" /> Abbildung 2. Monochoriale Plazenta in einem TTTS Schwangerschaft mit serieller amnioreduction zeigt das Vorhandensein von nur AV (weiße Sterne) und VA behandelt (grüne Sterne) Anastomosen ohne AA Anastomose. <img alt="Abbildung 3" src="/files/ftp_upload/3208/3208fig3.jpg" /> Abbildung 3. TTTS Plazenta nach fetoscopic Laserkoagulation der Gefäßanastomosen Verwendung des selektiven Laser-Technik, bei der ein kleiner Rest Anastomose versehentlich Patent wurde nach links (weißer Stern). Mit dem Selective Laser-Technik sind die Gefäßanastomosen zunächst identifiziert und anschließend koaguliert eins nach dem anderen. <img alt="Abbildung 4" src="/files/ftp_upload/3208/3208fig4.jpg" /> Abbildung 4. TTTS Plazenta nach fetoscopic Laserkoagulation mit der Solomon-Technik, bei dem nach Identifikation und Koagulation des einzelnen Anastomose, die komplette Gefäßäquator aus einer Plazenta-Marge auf die andere ist geronnen. <img alt="Abbildung 5" src="/files/ftp_upload/3208/3208fig5.jpg" /> Abbildung 5. TAPS Plazenta in das nur wenige, winzige VA Anastomosen (weiße Sterne) sind sichtbar entlang der vaskulären Äquator. <img alt="Abbildung 6" src="/files/ftp_upload/3208/3208fig6.jpg" /> Abbildung 6. Die mütterliche Seite der TAPS Plazenta (siehe Abbildung 5) zeigt die charakteristische Farbe Unterschied zwischen den beiden plazentaren Anteile. <img alt="Abbildung 7" src="/files/ftp_upload/3208/3208fig7.jpg" /> Abbildung 7. Monochoriale Plazenta einer Zwillingsschwangerschaft mit selektiver intrauterine Wachstumsretardierung eines Zwillings. Das Wachstum Fetus hat einen velamentous Nabelschnuransatz und eine viel kleinere Plazenta teilen. Der weiße Stern steht für eine AA-Anastomose. <img alt="Abbildung 8" src="/files/ftp_upload/3208/3208fig8.jpg" /> Abbildung 8 monoamniotische Plazenta:. Beachten Sie die verschiedenen AV (grüner Stern), VA (blauer Stern) und die 2 AA Anastomosen (weiße Sterne), und der kurze Abstand zwischen den beiden Kabel-Insertionen.

<ol>
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Pathol. 7, 159-165 (2004).</li><li>De Paepe, M. E., DeKoninck, P., Friedman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+distribution+patterns+in+monochorionic+twin+placentas.">Vascular distribution patterns in monochorionic twin placentas.</a> Placenta. 26, 471-475 (2005).</li><li>Nikkels, P. G., Hack, K. E., vanGemert, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathology+of+twin+placentas+with+special+attention+to+monochorionic+twin+placentas.">Pathology of twin placentas with special attention to monochorionic twin placentas.</a> J. Clin. Pathol. 61, 1247-1253 (2008).</li><li>Quintero, R. A., Martinez, J. 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TTTS und TAPS sind zwei schwere Erkrankungen, die in monochorialen Zwillingen auftreten kann. Beide Erkrankungen sind durch AV Anastomosen verursacht unausgewogen intertwin Durchblutung. In den letzten 2 Jahrzehnten wurde fetoscopic Laserchirurgie zur Verfügung und wurde gezeigt, dass die beste Methode zur Behandlung von TTTS 10 sein. Das Ziel der fetoscopic Laserbehandlung ist es, die inter-twin Zirkulation durch Koagulation des Gefäßanastomosen auf der Plazenta Oberfläche unterbrechen. Allerdings ist di...

<table border="1"><tbody><tr><th> Name des Reagenz </th><th> Firma </th><th> Katalog-Nummer </th><th> Kommentare </th></tr><tr><td> Umbilical Katheter (verschiedene Dimensionen, Länge 40 cm) </td><td> Vygon </td><td> 1270,08 (8 F) 1.270,08 (5 F) 1.270,04 (4 F) 1.270,03 (3,5 F) 1.270,02 (2,5 F) </td><td> Jede Art von Katheter verwendet werden kann </td></tr><tr><td> 20 ml Spritzen </td><td> BD Plastipak </td><td> 300613 </td><td></td></tr><tr><td> Farbe Farbstoff (verschiedene Farben) </td><td> Royal Talens Schoolverf </td><td> 36716010 (blau) 36715010 (grün) 36712350 (gelb) 36713570 (pink) </td><td> Jede viskosen Farbstoff verwendet werden kann </td></tr></tbody></table>

accurate and simple evaluation of vascular anastomoses in monochorionic placenta using colored dye

Die Anwesenheit von Plazenta Gefäßanastomosen ist eine conditio sine qua non für die Entwicklung von twin-to-twin Transfusion Syndrom (FFTS) und Twin-Anämie Polyzythämie Sequenz (TAPS) 1,2. Injection Studien twin Plazenten haben gezeigt, dass solche Anastomosen fast immer vorhanden sind, in monochorialen Zwillingen und äußerst selten in dichorialen Zwillinge 1. Drei Arten von Anastomosen wurden dokumentiert: von Arterie zu Arterie, von Vene zu Vene und Arterie zu Vene aus. Arterio-venöse (AV) Anastomosen sind unidirektional und werden als &quot;deep&quot; Anastomosen bezeichnet, da sie gehen durch eine gemeinsame Plazenta Keimblatt, während arterio-arterielle (AA) und veno-venöse (VV) Anastomosen bidirektionale sind und gemäß als &quot;oberflächlich&quot;, da sie auf der Chorionplatte liegen. Beide TTTS und TAPS werden von net Ungleichgewicht des Blutflusses zwischen den Zwillingen durch AV Anastomosen verursacht. Blut von einem Zwilling (Donor) ist durch eine Arterie in der gemeinsamen Plazenta cotyledon gepumpt und dann durch eine Vene in den Kreislauf des anderen Zwillings (der Empfänger) abgelassen. Es sei denn Blut ist wieder aus der Empfängerliste des Spenders durch gegenläufige tiefen AV Anastomosen oder durch oberflächliche Anastomosen gepumpt, tritt ein Ungleichgewicht von Blutvolumen, allmählich die zur Entwicklung von TTTS oder TAPS. Die Anwesenheit eines AA Anastomose hat sich gezeigt, gegen die Entwicklung von TTTS und TAPS durch Ausgleich für die Kreislauf-Ungleichgewicht durch die uni-direktionale AV Anastomosen 1,2 zu schützen. Die Injektion von monochorialen Plazenten bald nach der Geburt ist ein nützliches Mittel, um die Ätiologie der verschiedenen (hämatologischen) Komplikationen in monochorialen Zwillinge verstehen und ist eine erforderliche Prüfung, um die Diagnose von TAPS 2 zu erreichen. Darüber hinaus ermöglicht Injektion von TTTS Plazenten mit fetoscopic Laserchirurgie behandelt Identifizierung möglicher Rest-Anastomosen 3-5. Diese zusätzlichen Informationen sind von entscheidender Bedeutung für alle Perinatologen in der Verwaltung und Pflege von monochorialen Zwillingen mit FFTS oder TAPS beteiligt. Mehrere Plazenta Injektionstechniken werden derzeit verwendet. Wir bieten ein einfaches Protokoll zur genauen Beurteilung der Gegenwart von (Rest-) Gefäßanastomosen mit Farbstoff Injektion.

Watch this Scientific Journal Video about Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye at JoVE.com

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

Twin-to-twin Transfusion-Syndrom und zwei Anämie Polyzythämie Sequenz sind zwei potenziell verheerende Probleme in Perinatalmedizin. Beide Erkrankungen treten nur in monochorialen Zwillingen und resultieren aus unsymmetrischen Durchblutung der Plazenta Gefäßanastomosen. Wir bieten ein einfaches Protokoll zur genauen Beurteilung der Anwesenheit von Gefäßanastomosen mit Farbstoff Injektion von Plazentagefäße nach der Geburt.

Präzise und einfache Auswertung der Gefäßanastomosen in Monochoriale Placenta mit Farbstoff

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 ...

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26.6K Aufrufe.  Leiden University Medical Center. Twin-to-twin Transfusion-Syndrom und zwei Anämie Polyzythämie Sequenz sind zwei potenziell verheerende Probleme in Perinatalmedizin. Beide Erkrankungen treten nur in monochorialen Zwillingen und resultieren aus unsymmetrischen Durchblutung der Plazenta Gefäßanastomosen. Wir bieten ein einfaches Protokoll zur genauen Beurteilung der Anwesenheit von Gefäßanastomosen mit Farbstoff Injektion von Plazentagefäße nach der Geburt.

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

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&amp;#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.

Genaue und einfache Messung der pro-inflammatorischen Zytokins IL-1β mit einem Vollblut Stimulation Assay

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

Forschung

Medizin

Guide Wire Assisted Catheterization and Colored Dye Injection for Vascular Mapping of Monochorionic Twin Placentas

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC twin pregnancies develop complications arising from the shared placenta and associated vascular connections1. Severe twin-to-twin syndrome (TTTS) is reported to account for approximately 20% of these complications2,3. Inter-twin vascular connections occur in almost all MC placentas and are related to the prognosis and outcome of these high-risk twin pregnancies. The number, size and type of connections have been implicated in the development of TTTS and other MC twin conditions. Three types of inter-twin vascular connections occur: 1) artery to vein connections (AVs) in which a branch artery carrying deoxygenated blood from one twin courses along the fetal surface of the placenta and dives into a placental cotyledon. Blood flows via a deep intraparenchymal capillary network into a draining vein that emerges at the fetal surface of the placenta and brings oxygenated blood toward the other twin. There is unidirectional flow from the twin supplying the afferent artery toward the twin receiving the efferent vein; 2) artery to artery connections (AAs) in which a branch artery from each twin meets directly on the superficial placental surface resulting in a vessel with pulsatile bidirectional flow, and 3) vein to vein connections (VVs) in which a branch vein from each twin meets directly on the superficial placental surface allowing low pressure bidirectional flow. In utero obstetric sonography with targeted Doppler interrogation has been used to identify the presence of AV and AA connections4. Prenatally detected AAs that have been confirmed by postnatal placental injection studies have been shown to be associated with an improved prognosis for both twins5. Furthermore, fetoscopic laser ablation of inter-twin vascular connections on the fetal surface of the shared placenta is now the preferred treatment for early, severe TTTS. 
Postnatal placental injection studies provide a valuable method to confirm the accuracy of prenatal Doppler ultrasound findings and the efficacy of fetal laser therapy6. Using colored dyes separately hand-injected into the arterial and venous circulations of each twin, the technique highlights and delineates AVs, AAs, and VVs. This definitive demonstration of MC placental vascular anatomy may then be correlated with Doppler ultrasound findings and neonatal outcome to enhance our understanding of the pathophysiology of MC twinning and its sequelae. Here we demonstrate our placental injection technique.

Vascular mapping of monochorionic (MC) twin placentas after birth provides a means for detailed demonstration of vascular connections between the twins&#x2019; circulations. 
Imbalance of these connections is thought to play a pivotal role in the development of complications of MC twinning including twin-to-twin transfusion syndrome.

Führungsdraht Assisted Katheterisierung und Farbstoff Injection for Vascular Mapping von Monochoriale Twin Plazenten

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC ...

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

Eine einfache Methode der Maus Lung Intubation

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Technische Aspekte der Maus aortokavaler Fistel

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

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.

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.

Schnelle und genaue Messung der ausgeatmeten Luft Ammoniak

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

Evaluation of Tumor-infiltrating Leukocyte Subsets in a Subcutaneous Tumor Model

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or subvert anti-tumor immune responses in order to survive and flourish. Tumors take advantage of a number of different mechanisms of immune &#8220;escape,&#8221; including the recruitment of tolerogenic DC, immunosuppressive regulatory T cells (Tregs), and myeloid-derived suppressor cells (MDSC) that inhibit cytotoxic anti-tumor responses. Conversely, anti-tumor effector immune cells can slow the growth and expansion of malignancies: immunostimulatory dendritic cells, natural killer cells which harbor innate anti-tumor immunity, and cytotoxic T cells all can participate in tumor suppression. The balance between pro- and anti-tumor leukocytes ultimately determines the behavior and fate of transformed cells; a multitude of human clinical studies have borne this out. Thus, detailed analysis of leukocyte subsets within the tumor microenvironment has become increasingly important. Here, we describe a method for analyzing infiltrating leukocyte subsets present in the tumor microenvironment in a mouse tumor model. Mouse B16 melanoma tumor cells were inoculated subcutaneously in C57BL/6 mice. At a specified time, tumors and surrounding skin were resected en bloc and processed into single cell suspensions, which were then stained for multi-color flow cytometry. Using a variety of leukocyte subset markers, we were able to compare the relative percentages of infiltrating leukocyte subsets between control and chemerin-expressing tumors. Investigators may use such a tool to study the immune presence in the tumor microenvironment and when combined with traditional caliper size measurements of tumor growth, will potentially allow them to elucidate the impact of changes in immune composition on tumor growth. Such a technique can be applied to any tumor model in which the tumor and its microenvironment can be resected and processed.

This protocol describes a method for the detailed evaluation of leukocyte subsets within the tumor microenvironment in a mouse tumor model. Chemerin-expressing B16 melanoma cells were implanted subcutaneously into syngeneic mice. Cells from the tumor microenvironment were then stained and analyzed by flow cytometry, allowing for detailed leukocyte subset analyses.

Bewertung der Tumor-infiltrierenden Leukozyten-Untergruppen in einer subkutanen Tumormodell

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the aorta in mice using the sleeve method was short and easy averaging 20 min, permitting studies of iso/allo grafts. The following article describes the aortic transplantation procedure used in our laboratory. The mice were anesthetized with a mixture of 1.5% volume isoflurane and 100% oxygen through a face mask. At this point, the segment of the aorta between the renal arteries and its bifurcation was separated from the vena cava, freely prepared and clampedat the proximal and distal segments with a single silk suture. Prior to the removal of the aorta, a saline solution containing heparin was injected into the inferior vena cava. Then the aorta was cut between the clamps and a saline heparin solution was used to flush the lumen. The sleeve technique with monofilament sutures was used in order to transplant the abdominal aorta in the orthotopic position.

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

Mit Hilfe der Sleeve-Technik in einem Mausmodell der Aorta Transplantation - ein Lehr-Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

Model Surgical Training: Skills Acquisition in Fetoscopic Laser Photocoagulation of Monochorionic Diamniotic Twin Placenta Using Realistic Simulators

Fetoscopic laser coagulation of arterio-venous anastomoses (AVA) in a monochorionic placenta is the standard of care for twin-twin transfusion syndrome (TTTS), but is technically challenging and can lead to significant complications. Acquiring and maintaining the necessary surgical skills require consistent practice, a critical caseload, and time. Training on realistic surgical simulators can potentially shorten this steep learning curve and enables several proceduralists to acquire procedure-specific skills simultaneously. Here we describe realistic simulators designed to allow the user familiarity with the equipment and specific steps required in the surgical treatment of TTTS, including fetoscopic handling, approaches to anterior and posterior placenta, recognition of anastomoses, and efficient coagulation of vessels. We describe the skills that are especially important in conducting placental laser coagulation that the surgeon can practice on the model and apply in a clinical case. These models can be adapted easily depending on the availability of materials and require standard fetoscopy equipment. Such training systems are complementary to traditional surgical apprenticeships and can be useful aids for fetal medicine units that provide this clinical service.

Practicing the specific skills required for fetoscopic laser coagulation of monochorionic placental anastomoses on realistic models can aid less experienced surgeons in overcoming the steep learning curve associated with this procedure that is now regarded as the standard of care for twin-twin transfusion syndrome.

Modell chirurgische Ausbildung: Kompetenzentwicklung in Fetoscopic Laser Photokoagulation monochorialen dichorial Twin Plazenta mit realistischen Simulatoren

Fetoscopic laser coagulation of arterio-venous anastomoses (AVA) in a monochorionic placenta is the standard of care for twin-twin transfusion syndrome ...

Noninvasive Determination of Vortex Formation Time Using Transesophageal Echocardiography During Cardiac Surgery

Trans-mitral blood flow produces a three-dimensional rotational body of fluid, known as a vortex ring, that enhances the efficiency of left ventricular (LV) filling compared with a continuous linear jet. Vortex ring development is most often quantified with vortex formation time (VFT), a dimensionless parameter based on fluid ejection from a rigid tube. Our group is interested in factors that affect LV filling efficiency during cardiac surgery. In this report, we describe how to use standard two-dimensional (2D) and Doppler transesophageal echocardiography (TEE) to noninvasively derive the variables needed to calculate VFT. We calculate atrial filling fraction (&#946;) from velocity-time integrals of trans-mitral early LV filling and atrial systole blood flow velocity waveforms measured in the mid-esophageal four-chamber TEE view. Stroke volume (SV) is calculated as the product of the diameter of the LV outflow track measured in the mid-esophageal long axis TEE view and the velocity-time integral of blood flow through the outflow track determined in the deep transgastric view using pulse-wave Doppler. Finally, mitral valve diameter (D) is determined as the average of major and minor axis lengths measured in orthogonal mid-esophageal bicommissural and long axis imaging planes, respectively. VFT is then calculated as 4 &#215; (1-&#946;) &#215; SV/(&#960;D3). We have used this technique to analyze VFT in several groups of patients with differing cardiac abnormalities. We discuss our application of this technique and its potential limitations and also review our results to date. Noninvasive measurement of VFT using TEE is straightforward in anesthetized patients undergoing cardiac surgery. The technique may allow cardiac anesthesiologists and surgeons to assess the impact of pathological conditions and surgical interventions on LV filling efficiency in real time.

We describe a protocol to measure vortex formation time, an index of left ventricular filling efficiency, using standard transesophageal echocardiography techniques in patients undergoing cardiac surgery. We apply this technique to analyze vortex formation time in several groups of patients with differing cardiac pathologies.

Nicht-invasive Zeitbestimmung Vortex Formation mit transösophagealen Echokardiographie bei Herzchirurgie

Trans-mitral blood flow produces a three-dimensional rotational body of fluid, known as a vortex ring, that enhances the efficiency of left ventricular ...

Echocardiographic Evaluation of Atrial Communications before Transcatheter Closure

Transthoracic (TTE) and transesophageal echocardiography (TEE) is the standard imaging method for atrial septal defect (ASD) and patent foramen ovale (PFO) detection, for patient selection for transcatheter ASD/PFO closure,&#160;for intraoperative guidance and for long-term follow-up. The size, shape, location and the number of the atrial communications schould be determined. The accuracy of PFO detection can be improved by using agitated saline together with maneuvers to transiently increase the right atrial (RA) pressure. The appearance of microbubbles in the left atrium (LA) within 3 cardiac cycles after opacification of the RA is considered positive for the presence of an intracardiac shunt. Three dimensional TEE identifies further septal fenestrations and describes the dynamic morphology of ASD/PFO and atrial septal aneurysm. Follow-up evaluations with TTE is recommended at 1, 6, and 12 months after the procedure, with a subsequent evaluation every year. Previous studies showed an increased incidence of atrial arrhythmias early after device closure. Speckle tracking analysis may help to understand functional left atrial remodeling following percutaneous closure and its impact on atrial arrhythmias.

Transthoracic (TTE) and transesophageal (TEE) echocardiography represent the basic imaging tools for interatrial septum examination. Three dimensional (3D) TEE provides incremental information in the assessment of the interatrial septum. Further advanced echocardiography techniques using speckle tracking echocardiography are applied for sensitive volumetric and functional assessment of the heart chambers.&#160;

Echokardiographische Auswertung der Atrialkommunikation vor dem Transkatheterverschluss

Transthoracic (TTE) and transesophageal echocardiography (TEE) is the standard imaging method for atrial septal defect (ASD) and patent foramen ovale ...