Le financement de la publication est assurée par Cerus Corp, le fabricant du système sanguin INTERCEPT.

<ol>
	<li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide to the preparation, use and quality assurance of blood components. , (2010).</li><li>Van Rhenen, D. J., Vermeij, J., Mayaudon, V., 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=Functional+characteristics+of+S-59+photochemically+treated+platelet+concentrates+derived+from+buffy+coats.">Functional characteristics of S-59 photochemically treated platelet concentrates derived from buffy coats.</a> Vox Sang. 79, 206-214 (2000).</li><li>Wollowitz, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+DNA+and+RNA+in+pathogens:+mode+of+action+of+amotosalen+HCl.">Targeting DNA and RNA in pathogens: mode of action of amotosalen HCl.</a> Transfus. Med. Rev. 31, 11-16 (2004).</li><li>Irsch, J., Lin, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogen+Inactivation+of+Platelet+and+Plasma+Blood+Components+for+Transfusion+Using+the+INTERCEPT+Blood+SystemTM.">Pathogen Inactivation of Platelet and Plasma Blood Components for Transfusion Using the INTERCEPT Blood SystemTM.</a> Transfus. Med. Hemother. 38, (2011).</li><li>Lin, L., Dikeman, R., Molini, B., 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=Photochemical+treatment+of+platelet+concentrates+with+amotosalen+and+long-wavelength+ultraviolet+light+inactivates+a+broad+spectrum+of+pathogenic+bacteria.">Photochemical treatment of platelet concentrates with amotosalen and long-wavelength ultraviolet light inactivates a broad spectrum of pathogenic bacteria.</a> Transfusion. 44, 1496-1504 (2004).</li><li>Lin, L., Hanson, C., Alter, H., 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=Inactivation+of+viruses+in+platelet+concentrates+by+photochemical+treatment+with+amotosalen+and+long-wavelength+ultraviolet+light.">Inactivation of viruses in platelet concentrates by photochemical treatment with amotosalen and long-wavelength ultraviolet light.</a> Transfusion. 45, 580-590 (2005).</li><li>Allain, J. P., Cianco, C., Blajchman, A., 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=Protecting+the+blood+supply+from+emerging+pathogens:+the+role+of+pathogen+inactivation.">Protecting the blood supply from emerging pathogens: the role of pathogen inactivation.</a> Transfus. Med. Rev. 19, 110-126 (2005).</li><li>Stramer, S., Hollinger, F., Katz, L., 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=Emerging+infectious+disease+agents+and+their+potential+threat+to+transfusion+safety.">Emerging infectious disease agents and their potential threat to transfusion safety.</a> Transfusion. 49, 1S-29S (2009).</li><li>Nussbaumer, W., Allesdorfer, D., Grabmer, 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=Prevention+of+transfusion+of+platelet+components+contaminated+with+low+levels+of+bacteria:+a+comparison+of+bacteria+culture+and+pathogen+inactivation+methods.">Prevention of transfusion of platelet components contaminated with low levels of bacteria: a comparison of bacteria culture and pathogen inactivation methods.</a> Transfusion. 47, 1125-1133 (2007).</li><li>Schlenke, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protection+against+Transfusion-Associated+Graft-versus-Host+Disease+in+Blood+transfusion:+Is+Gamma-Irradiation+the+Only+Answer?+Transfus.">Protection against Transfusion-Associated Graft-versus-Host Disease in Blood transfusion: Is Gamma-Irradiation the Only Answer? Transfus.</a> Med. Hemother. 31, 24-31 (2004).</li><li>Lin, L., Corash, L., Osselear, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protection+Against+TA-GVHD+Due+to+Platelet+Transfusion+By+Using+Pathogen+Inactivation+with+the+INTERCEPT+Blood+SystemTM+-+Gamma+Irradiation+is+Not+the+Only+Answer.">Protection Against TA-GVHD Due to Platelet Transfusion By Using Pathogen Inactivation with the INTERCEPT Blood SystemTM - Gamma Irradiation is Not the Only Answer.</a> Haematologica. 95 (Extra 1), 230-237 (2010).</li><li>Corash, L., Lin, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+processes+for+inactivation+of+leukocytes+to+prevent+transfusion-associated+graft-verus-host+disease.">Novel processes for inactivation of leukocytes to prevent transfusion-associated graft-verus-host disease.</a> Bone Marrow Transplant. 33, 1-7 (2004).</li></ol>

Production et le libre accès à cet article est sponsorisé par Cerus.

Couche leuco-plaquettaire nécessitent plusieurs étapes de traitement qui résultent en post-collecte coûts, y compris le temps du personnel, les consommables, l&#39;équipement et les déchets, qui doit être pris en compte dans le coût global de la production des unités de plaquettes. Amélioration du rendement des plaquettes de chaque couche leuco-plaquettaire (via l&#39;optimisation du volume de la couche leucocytaire et de l&#39;hématocrite) permet la production d&#39;une unité à double couche leuco-plaquettaire dose de plaquettes à partir d&#39;un pool de couches leuco-plaquettaires sept. Lorsque cette optimisation est réalisée, le nombre de couches leucocytaires nécessaire pour produire un nombre fixe de doses de plaquettes peut être réduite, ce qui améliore l&#39;utilisation globale de la couche leuco et permettant la production de plaquettes supplémentaires de concentrés (PC). Temps du personnel ainsi que les coûts de consommables et d&#39;équipements tels que des ensembles de mise en commun, la solution additive plaquettaire, plaquettes de connexion stériles, ensembles de filtre, et les procédures de centrifugation sont réduites jusqu&#39;à 50%. En plus d&#39;optimiser la production de plaquettes et de réduire les coûts associés, this la mise en commun méthode peut générer un concentré de plaquettes (CP) qui répond aux exigences d&#39;entrée pour une utilisation avec l&#39;ensemble INTERCEPT de stockage à double traitement des conteneurs, ce qui nous permet d&#39;offrir une meilleure protection à nos receveurs de transfusions aussi bien. Le système sanguin INTERCEPT pour l&#39;inactivation des agents pathogènes utilise amotosalen et les rayons ultraviolets A (UVA) de façon covalente réticuler l&#39;ADN et de l&#39;ARN, la prévention de réplication des acides nucléiques et des agents pathogènes rendu incapables de provoquer une maladie. 3 Il inactive efficacement un large spectre d&#39;agents pathogènes tels que les virus, les bactéries , les parasites et les contaminants cellules de sang blanches. 4-6 INTERCEPT fournit une alternative au paradigme actuel tests pour les pathogènes émergents, ce qui implique historiquement un retard considérable alors qu&#39;un nouveau test est développé et exige finalement un investissement financier important pour mettre en œuvre lors d&#39;un test devient disponible. 7,8 Il peut également fournir une alternative aux mesures de sécu redondanteres telles que la détection bactérienne 9 et irradiation gamma. 10-12 En outre, INTERCEPT nous permet de traiter doubles unités de plaquettes dose qui profite à l&#39;efficacité de notre production et nous aide à respecter nos budgets. INTERCEPT et la méthode de double dose couche leuco-plaquettaire peut être adapté à une multitude de tâches de sang du centre. À titre d&#39;exemples, collectes de sang total peut être augmenté à 500 ml; collections pourraient aussi être stockée pendant la nuit à 22 ° C avant la séparation. En outre, un autre procédé de mise en commun de la couche leucocytaire est possible (par exemple un procédé de poulpe à la place d&#39;un procédé de train) et / ou un dispositif automatique (par exemple TACSI) peuvent être utilisés pour la deuxième centrifugation et séparation de la suspension de plaquettes à partir des cellules restantes rouges . Si augmentation de la teneur en plaquettes est nécessaire, plusieurs techniques sont disponibles, y compris de pré-sélection des donneurs sur la base de la numération plaquettaire, une nuit d&#39;incubation du sang total, adjustement des paramètres de centrifugation, ou en utilisant un pool de couches leuco-plaquettaires 8 au lieu de 7. En raison de la limitation de la mise en commun établit actuellement disponible et la capacité des pots à centrifuger, le volume total de la piscine de la couche leucocytaire doit être d&#39;environ 600 ml. Le processus de centrifugation et les paramètres décrits dans le présent protocole ont été optimisés pour obtenir un produit qui répond aux paramètres INTERCEPT pour un pool de couches leuco-plaquettaires 7. Les paramètres de centrifugation doit être modifié si le nombre de couches leucocytaires dans la piscine est modifié. Dans certains pays, la dose minimale des plaquettes est plus élevé et les exigences de CQ sont plus strictes que dans la Suède. Ainsi, nos résultats ne sont pas universellement applicables. Dans ces pays, le PC double dose avant le traitement INTERCEPT devra contenir un plus grand nombre de plaquettes afin de répondre aux exigences locales après traitement INTERCEPT et split. Parce que la teneur maximale des plaquettes et volume de traitement for INTERCEPT sont 7x10 11 plaquettes et 420 ml respectivement, le pourcentage d&#39;unités qui dépassent les exigences de traitement INTERCEPT ou n&#39;ont pas suffisamment de contenu plaquettaire être divisée en deux doses thérapeutiques, varieront en fonction de facteurs tels que les exigences locales de dose de plaquettes, les critères QC, résultats buffy coat optimisation et la stabilité de la production. Si un PC double dose dépasse les exigences de traitement INTERCEPT, il peut être ajustée manuellement pour répondre aux exigences et ultérieurement traitées. Dans les rares occasions où le pool de couches leuco-7-plaquettes rendements insuffisants pour un produit double dose après INTERCEPT, nous choisissons de ne pas effectuer le traitement INTERCEPT. Alternativement, d&#39;autres centres de transfusion peuvent choisir de traiter le PC avec un ensemble de traitement INTERCEPT dose unique (c&#39;est à dire l&#39;ensemble de grand volume) et stocker le PC en une seule dose thérapeutique plus large, réalisant ainsi l&#39;inactivation des pathogènes sur toutes les unités. Afin de nous assurer de répondre à notreExigences et aux agents pathogènes QC inactiver le plus grand nombre de nos unités de plaquettes que possible, nous sélectionnons couches leucocytaires basées sur les comptes des bailleurs de fonds de plaquettes, qui, une fois regroupés, se traduira par une teneur minimale des plaquettes de 5.6x10 11 plaquettes dans la piscine. Cela garantit suffisamment le contenu des plaquettes pour produire deux doses thérapeutiques après traitement INTERCEPT. Notre validation démontre que 7 unités dérivées de sang total couche leuco-plaquettaire peuvent être mises en commun et avec succès traité par le procédé INTERCEPT pour les plaquettes, résultant en 2 pathogènes inactivés produits plaquettaires qui répondent aux critères d&#39;acceptation pour la fabrication (suédois exigences et les lignes directrices européennes) et pour le soutien des patients nécessitant des transfusions de plaquettes selon les lignes directrices de pratique clinique et les méthodes standards de perfusion de plaquettes en Suède.

<table border="1">
	<tbody>
		<tr>
			<td>Material / Equipment</td>
			<td>Company</td>
			<td>Catalogue Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Whole blood donation, primary separation, and platelet production</td>
		</tr>
		<tr>
			<td>Blood collection pack</td>
			<td>Fenwal</td>
			<td>R6485</td>
			<td>Top/Bottom set</td>
		</tr>
		<tr>
			<td>Automated component extractor</td>
			<td>Fenwal</td>
			<td>Optipress-II</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Blood mixer and balance system</td>
			<td>Baxter</td>
			<td>Easymix V3</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Platelet leukocyte filtration set</td>
			<td>Fenwal</td>
			<td>K4R7042</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Hettich</td>
			<td>Roto Silenta 63 RS</td>
			<td>Version 5.5</td>
		</tr>
		<tr>
			<td>Platelet additive solution - SSP+</td>
			<td>MacoPharma</td>
			<td>SSP2030U</td>
			<td>300 ml</td>
		</tr>
		<tr>
			<td>Sterile tubing welder</td>
			<td>Terumo</td>
			<td>T-SCD</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>INTERCEPT treatment &amp; storage</td>
		</tr>
		<tr>
			<td>INTERCEPT processing set</td>
			<td>Cerus</td>
			<td>INT2503</td>
			<td>Dual Storage (DS) set</td>
		</tr>
		<tr>
			<td>INTERCEPT Illuminator</td>
			<td>Cerus</td>
			<td>INT100</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>PC sample pack</td>
			<td>Fenwal</td>
			<td>FTX 1122</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Incubator</td>
			<td>Helmer</td>
			<td>PC2200/PC3200</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Agitator</td>
			<td>Helmer</td>
			<td>PF48H/PF96H</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Evaluation of in vitro Platelet Function</td>
		</tr>
		<tr>
			<td>Blood gas analyzer</td>
			<td>Radiometer</td>
			<td>ABL 735</td>
			<td>Used for pH, blood gases, and lactate measurement</td>
		</tr>
		<tr>
			<td>Chemistry system</td>
			<td>Ortho Clinical Diagnostic</td>
			<td>Vitros 5.1</td>
			<td>Used for glucose measurement</td>
		</tr>
		<tr>
			<td>Hematology analyzer</td>
			<td>Boule Medical AB</td>
			<td>Medonic CA620-Cellguard</td>
			<td>Used for platelet count measurement</td>
		</tr>
		<tr>
			<td>Flow cytometer</td>
			<td>BD</td>
			<td>FACSCanto</td>
			<td>Used for white blood cell measurement</td>
		</tr>
	</tbody>
</table>

preparation and pathogen inactivation of double dose buffy coat platelet products using the intercept blood system

Les centres de transfusion sont confrontés à de nombreux défis, y compris le rendement de production de maximiser les dons de produits sanguins qu&#39;ils reçoivent ainsi que d&#39;assurer le plus haut niveau de sécurité possible pour les patients transfusés, y compris la protection contre les maladies transmissibles par transfusion. Cela doit être fait d&#39;une manière financièrement responsable qui minimise les frais d&#39;exploitation, y compris les consommables, les équipements, les déchets et les coûts de personnel, entre autres. Plusieurs méthodes sont disponibles pour produire des concentrés de plaquettes pour transfusion. Un des plus commune est la méthode de la couche leuco-plaquettaire dans laquelle une seule unité thérapeutique de plaquettes (≥ 2,0 x10 11 plaquettes par unité ou par les réglementations locales) est préparé par la mise en commun de la couche leuco-plaquettaire à partir de six dons de sang total. Un procédé de production de &quot;double dose&quot; plaquettes dérivées du sang total n&#39;a été que récemment mis au point. Présenté ici est un nouveau procédé de préparationdeux plaquettes sanguines dose entière concentrés dérivés de pools de couches leucocytaires 7 et ensuite traiter les unités de dose double avec INTERCEPT Blood System pour l&#39;inactivation des agents pathogènes. INTERCEPT a été développé pour inactiver les virus, les bactéries, les parasites et les cellules contaminantes donateurs blancs qui peuvent être présents dans les dons de sang. Jumelage avec INTERCEPT la méthode de la couche leuco-plaquettaire double dose en utilisant le traitement INTERCEPT Set avec Conteneurs de stockage doubles (le «jeu DS&quot;), permet aux centres sanguins pour traiter chacune de leurs unités de dose double en une seule série de traitement d&#39;inactivation des pathogènes, maximisant ainsi la sécurité des patients tout en minimisant les coûts. La méthode de double dose couche leuco-plaquettaire nécessite moins de couches leuco-plaquettaires et réduit l&#39;utilisation de consommables de plus de 50% (par exemple, la mise en commun des ensembles, ensembles de filtre, la solution additive plaquettaire, et gaufrettes connexion stérile) par rapport à la préparation et au traitement des unités à dose unique couche leuco-plaquettaire . Autres économies comprennent moins de déchets, moins d&#39;équipement Maintenance, demande en énergie, réduction du temps de personnel et les coûts de collecte inférieure par rapport à la technique d&#39;aphérèse.

Watch this Scientific Journal Video about Preparation and Pathogen Inactivation of Double Dose Buffy Coat Platelet Products using the INTERCEPT Blood System at JoVE.com

Preparation and Pathogen Inactivation of Double Dose Buffy Coat Platelet Products using the INTERCEPT Blood System

Cet article décrit le processus utilisé par Örebro University Hospital pour produire des plaquettes à double couche leuco-plaquettaire dose des concentrés préparés à partir de dons de sang total et traitées avec INTERCEPT Blood System pour l&#39;inactivation des agents pathogènes. La<em&gt; In vitro</em&gt; La qualité des unités finales plaquettes sont évaluées sur 7 jours de stockage.

Préparation et inactivation des agents pathogènes des doubles produits Buffy Coat dose de plaquettes à l&#39;aide du système sanguin INTERCEPT

Blood centers are faced with many challenges including maximizing production yield from the blood product donations they receive as well as ensuring the ...

preparation-pathogen-inactivation-double-dose-buffy-coat-platelet

Research

JoVE Journal

Medicine

28.5K vues.  Örebro University Hospital. Cet article décrit le processus utilisé par Örebro University Hospital pour produire des plaquettes à double couche leuco-plaquettaire dose des concentrés préparés à partir de dons de sang total et traitées avec INTERCEPT Blood System pour l'inactivation des agents pathogènes. La In vitro La qualité des unités finales plaquettes sont évaluées sur 7 jours de stockage.

Vidéo: Preparation and Pathogen Inactivation of Double Dose Buffy Coat Platelet Products using the INTERCEPT Blood System

Minimal Erythema Dose (MED) Testing

Ultraviolet radiation (UV) therapy is sometimes used as a treatment for various common skin conditions, including psoriasis, acne, and eczema. The dosage of UV light is prescribed according to an individual's skin sensitivity. Thus, to establish the proper dosage of UV light to administer to a patient, the patient is sometimes screened to determine a minimal erythema dose (MED), which is the amount of UV radiation that will produce minimal erythema (sunburn or redness caused by engorgement of capillaries) of an individual's skin within a few hours following exposure. This article describes how to conduct minimal erythema dose (MED) testing. There is currently no easy way to determine an appropriate UV dose for clinical or research purposes without conducting formal MED testing, requiring observation hours after testing, or informal trial and error testing with the risks of under- or over-dosing. However, some alternative methods are discussed.

Minimal Erythema Dose MED Testing

This article describes how to conduct minimal erythema dose (MED) testing in order to determine the lowest dose of ultraviolet radiation that will cause erythema (burning) when administered to an individual.

Erythème Dose (MED) des tests minimaux

Ultraviolet radiation (UV) therapy is  sometimes used as a treatment for various common skin conditions, including  psoriasis, acne, and eczema. The ...

Recherche

Médecine

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia) 1. Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex 2, is the most common site of human stroke 3, and ischemia within its territory can result in extensive dysfunction or death 1,4,5. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA.
Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

We describe a highly reproducible method for the permanent occlusion of a rodent major cerebral blood vessel. This technique can be accomplished with very little peripheral damage, minimal blood loss, a high rate of long-term survival, and consistent infarct volume commensurate with the human clinical population.

Permanent occlusion des vaisseaux cérébraux<em&gt; Via</em&gt; Double ligature et Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

Drug Repurposing Hypothesis Generation Using the "RE:fine Drugs" System

The promise of drug repurposing is that existing drugs may be used for new disease indications in order to curb the high costs and time for approval. The goal of computational methods for drug repurposing is to enable solutions for safer, cheaper and faster drug discovery. Towards this end, we developed a novel method that integrates genetic and clinical phenotype data from large-scale GWAS and PheWAS studies with detailed drug information on the concept of transitive Drug-Gene-Disease triads. We created "RE:fine Drugs," a freely available, interactive dashboard that automates gene, disease and drug-based searches to identify drug repurposing candidates. This web-based tool supports a user-friendly interface that includes an array of advanced search and export options. Results can be prioritized in a variety of ways, including but not limited to, biomedical literature support, strength and statistical significance of GWAS and/or PheWAS associations, disease indications and molecular drug targets. Here we provide a protocol that illustrates the functionalities available in the "RE:fine Drugs" system and explores the different advanced options through a case study.

Drug Repurposing Hypothesis Generation Using the &quot;RE:fine Drugs&quot; System

Here we describe a protocol using the web-based drug repurposing hypothesis generation tool: "RE:fine Drugs." This protocol can be modified to a user's preferences at the level of the query type (gene, drug or disease) and/or the range of available advanced options.

Système: Drug Repurposing Hypothesis Generation Utilisation de la &quot;Drogues fines RE&quot;

The promise of drug repurposing is that existing drugs may be used for new disease indications in order to curb the high costs and time for approval. The ...

Turbidimetry on Human Washed Platelets: The Effect of the Pannexin1-inhibitor Brilliant Blue FCF on Collagen-induced Aggregation

Turbidimetry is a laboratory technique that is applied to measure the aggregation of platelets suspended in either plasma (platelet-rich plasma, PRP) or in buffer (washed platelets), by the use of one or a combination of agonists. The use of washed platelets separated from their plasma environment and in the absence of anticoagulants allows for studying intrinsic platelet properties. Among the large panel of agonists, arachidonic acid (AA), adenosine di-phosphate (ADP), thrombin and collagen are the most frequently used. The aggregation response is quantified by measuring the relative optical density (OD) over time of platelet suspension under continuous stirring. Platelets in homogeneous suspension limit the passage of light after the addition of an agonist, platelet shape change occurs producing a small transitory increase in OD. Following this initial activation step, platelet clots form gradually, allowing the passage of light through the suspension as a result of decreased OD. The aggregation process is ultimately expressed as a percentage, compared to the OD of platelet-poor plasma or buffer. Rigorous calibration is thus essential at the beginning of each experiment. As a general rule: calibration to 0% is set by measuring the OD of a non-stimulated platelet suspension while measuring the OD of the suspension medium containing no platelets represents a value of 100%. Platelet aggregation is generally visualized as a real-time aggregation curve. Turbidimetry is one of the most commonly used laboratory techniques for the investigation of platelet function and is considered as the historical gold standard and used for the development of new pharmaceutical agents aimed at inhibiting platelet aggregation. Here, we describe detailed protocols for 1) preparation of human washed platelets and 2) turbidimetric analysis of collagen-induced aggregation of human washed platelets pretreated with the food dye Brilliant Blue FCF that was recently identified as an inhibitor of Pannexin1 (Panx1) channels.

We describe a straightforward method for the isolation of washed platelets from human blood followed by agonist-induced platelet aggregation measurements by turbidimetry. As an example we apply this method for studying the aggregation response of human platelets to collagen after a pre-incubation with the Pannexin1 channel inhibitor Brilliant Blue FCF.

Turbidimétrie sur Human Washed: L&#39;effet Plaquettes du Pannexin1 inhibiteur bleu brillant FCF sur l&#39;agrégation induite par le collagène

Turbidimetry is a laboratory technique that is applied to measure the aggregation of platelets suspended in either plasma (platelet-rich plasma, PRP) or ...

Routine Screening Method for Microparticles in Platelet Transfusions

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative for hospital blood banks. Cells fragment off microparticles (MP) when they are stressed. Blood and blood components may contain cellular fragments from a variety of cells, most notably from activated platelets. When performing their roles as innate immune cells and major players in coagulation and hemostasis, platelets change shape and generate microparticles. With dynamic light scattering (DLS)-based microparticle detection, it is possible to differentiate activated (high microparticle) from non-activated (low microparticle) platelets in transfusions, and optimize the use of this scarce blood product. Previous research suggests that providing non-activated platelets for prophylactic use in hematology-oncology patients could reduce their risk of becoming refractory and improve patient care. The goal of this screening method is to routinely differentiate activated from non-activated platelets. The method described here outlines the steps to be performed for routine platelet inventory management in a hospital blood bank: obtaining a sample from a platelet transfusion, loading the sample into the capillary for DLS measurement, performing the DLS test to identify microparticles, and using the reported microparticle content to identify activated platelets.

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative in hospital blood banks. The goal is to differentiate activated from non-activated platelets to optimize the use of platelets. Providing non-activated platelets to hematology-oncology patients might reduce their high risk to become refractory.

Routine de procédé de criblage de microparticules dans les Transfusions de plaquettes

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative for hospital blood ...

Preparation, Procedures and Evaluation of Platelet-Rich Plasma Injection in the Treatment of Knee Osteoarthritis

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department. Existing non-surgical treatments have a limited effect on the repair of cartilage and on bone regeneration. Platelet-rich plasma (PRP) is an autologous bioactive substance that can repair cartilage injury and accelerate bone regeneration effectively. However, reporting of PRP preparation protocols in clinical studies is highly inconsistent, with the majority of studies providing insufficient information to allow the protocol to be reproduced. We describe a repeatable method of preparing PRP visually, the treatment of KOA using PRP intra-articular injection, and methods of evaluating the outcome. PRP was prepared using manual double centrifugation. The PRP layer was extracted from peripheral blood and used for knee joint cavity injection. Evaluations included assessments of blood platelet concentrations and clinical outcomes. Preparation of PRP by manual centrifugation requires less apparatus and is less costly than plasma filtration or centrifugation using equipment. The centrifugation time of our double centrifugation method was 6 and 5 minutes for the respective centrifugations at forces of 800 and 1400 x g, respectively, to allow for the consistent preparation of standardized PRP. However, a manual method is susceptible to operator error, and PRP batch preparation is not available. Intra-articular injection of PRP proved to be an effective treatment for knee osteoarthritis. The entire treatment procedure took less than 30 minutes, the blood platelet concentration of PRP could be standardized, and treatment was proven to be effective when evaluated by follow-up.

Knee osteoarthritis is frequently seen in the orthopedic department. We introduce in detail the entire knee osteoarthritis treatment process with platelet-rich plasma injection, including preparation, procedures, and evaluation.

Préparation, procédures et l’évaluation de l’Injection de Plasma riche en plaquettes dans le traitement de l’arthrose du genou

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department. Existing non-surgical treatments have a limited ...

An In Vivo Method for Evaluating the Gut-Blood Barrier and Liver Metabolism of Microbiota Products

The gut-blood barrier (GBB) controls the passage of nutrients, bacterial metabolites and drugs from intestinal lumen to the bloodstream. The GBB integrity is disturbed in gastrointestinal, cardiovascular and metabolic diseases, which may result in easier access of biologically active compounds, such as gut bacterial metabolites, to the bloodstream. Thus, the permeability of the GBB may be a marker of both intestinal and extraintestinal diseases. Furthermore, the increased penetration of bacterial metabolites may affect the functioning of the entire organism.
Commonly used methods for studying the GBB permeability are performed ex vivo. The accuracy of those methods is limited, because the functioning of the GBB depends on intestinal blood flow. On the other hand, commonly used in vivo methods may be biased by liver and kidney performance, as those methods are based on evaluation of urine or/and peripheral blood concentrations of exogenous markers. Here, we present a direct measurement of GBB permeability in rats using an in vivo method based on portal blood sampling, which preserves intestinal blood flow and is virtually not affected by the liver and kidney function. 
Polyurethane catheters are inserted into the portal vein and inferior vena cava just above the hepatic veins confluence. Blood is sampled at baseline and after administration of a selected marker into a desired part of the gastrointestinal tract. Here, we present several applications of the method including (1) evaluation of the colon permeability to TMA, a gut bacterial metabolite, (2) evaluation of liver clearance of TMA, and (3) evaluation of a gut-portal blood-liver-peripheral blood pathway of gut bacteria-derived short-chain fatty acids. Furthermore, the protocol may also be used for tracking intestinal absorption and liver metabolism of drugs or for measurements of portal blood pressure.

An In Vivo Method for Evaluating the Gut-Blood Barrier and Liver Metabolism of Microbiota Products

The access of nutrients, microbiota metabolites and medicines to the circulation is controlled by the gut-blood barrier (GBB). We describe a direct method for measuring the GBB permeability in vivo, which, in contrast to commonly used indirect methods, is virtually not affected by liver and kidney functions.

Une méthode In Vivo pour l’évaluation de la barrière intestinale-sang et le métabolisme hépatique du microbiote produits

The gut-blood barrier (GBB) controls the passage of nutrients, bacterial metabolites and drugs from intestinal lumen to the bloodstream. The GBB integrity ...

Platelet-based Detection of Nitric Oxide in Blood by Measuring VASP Phosphorylation

Platelets are the blood components responsible for proper blood clotting. Their function is highly regulated by various pathways. One of the most potent vasoactive agents, nitric oxide (NO), can also act as a powerful inhibitor of platelet aggregation. Direct NO detection in blood is very challenging due to its high reactivity with cell-free hemoglobin that limits NO half-life to the millisecond range. Currently, NO changes after interventions are only estimated based on measured changes of nitrite and nitrate (members of the nitrate-nitrite-NO metabolic pathway). However precise, these measurements are rather difficult to interpret vis a vis actual NO changes, due to the naturally high baseline nitrite and nitrate levels that are several orders of magnitude higher than expected changes of NO itself. Therefore, the development of direct and simple methods that would allow one to detect NO directly is long overdue. This protocol addresses a potential use of platelets as a highly sensitive NO sensor in blood. It describes initial platelet rich plasma (PRP) and washed platelet preparations and the use of nitrite and deoxygenated red blood cells as NO generators. Phosphorylation of VASP at serine 239 (P-VASPSer239) is used to detect the presence of NO. The fact that VASP protein is highly expressed in platelets and that it is rapidly phosphorylated when NO is present leads to a unique opportunity to use this pathway to directly detect NO presence in blood.

Here, we present a protocol to address the potential use of platelets as a highly sensitive nitric oxide sensor in blood. It describes initial platelet preparation and the use of nitrite and deoxygenated red blood cells as nitric oxide generators.

Détection du monoxyde d’azote dans le sang en mesurant Phosphorylation VASP plaquettaire

Platelets are the blood components responsible for proper blood clotting. Their function is highly regulated by various pathways. One of the most potent ...

Measuring the Effects of Bacteria and Chemicals on the Intestinal Permeability of Caenorhabditis elegans

In living organisms, intestinal hyperpermeability is a serious symptom that leads to many inflammatory bowel diseases (IBDs). Caenorhabditis elegans is a nonmammalian animal model that is widely used as an assay system due to its short lifespan, transparency, cost-effectiveness, and lack of animal ethics issues. In this study, a method was developed to investigate the effects of different bacteria and 3,3'-diindolylmethane (DIM) on the intestinal permeability of C. elegans with a high-throughput image analysis system. The worms were infected with different gut bacteria or cotreated with DIM for 48 h and fed with fluorescein isothiocyanate (FITC)-dextran overnight. Then, the intestinal permeability was examined by comparing the fluorescence images and the fluorescence intensity inside the worm bodies. This method may also have the potential to identify probiotic and pathogenic intestinal bacteria that affect intestinal permeability in the animal model and is effective for examining the effects of harmful or health-promoting chemicals on intestinal permeability and intestinal health. However, this protocol also has some considerable limitations at the genetic level, especially for determining which genes are altered to control illness, because this method is mostly used for phenotypic determination. In addition, this method is limited to determining exactly which pathogenic substrates cause inflammation or increase the permeability of the worms' intestines during infection. Therefore, further in-depth studies, including investigation of the molecular genetic mechanism using mutant bacteria and nematodes as well as chemical component analysis of bacteria, are required to fully evaluate the function of bacteria and chemicals in determining intestinal permeability.

Measuring the Effects of Bacteria and Chemicals on the Intestinal Permeability of Caenorhabditis elegans

This protocol describes how to measure intestinal permeability of Caenorhabditis elegans. This method is helpful for basic biological research on intestinal health related to the interaction between intestinal bacteria and their host and for screening to identify probiotic and chemical agents to cure leaky gut syndrome and inflammatory bowel diseases.

Mesurer les effets des bactéries et des produits chimiques sur la perméabilité intestinale de Caenorhabditis elegans

In living organisms, intestinal hyperpermeability is a serious symptom that leads to many inflammatory bowel diseases (IBDs). Caenorhabditis elegans is a ...

Laminectomy for the Removal of Thoracic Ossification of the Ligamentum Flavum (TOLF) Using Ultrasonic and Conventional Osteotomes

Thoracic ossification of the ligamentum flavum (TOLF) is a common cause of progressive thoracic myelopathy. TOLF is typically treated with surgical decompression. A variety of surgical techniques, including laminoplasty, laminectomy, and lamina fenestration, are used for the effective treatment of TOLF. However, traditional methods are associated with a substantial risk of perioperative complications, including dural laceration and/or iatrogenic spinal cord injury. Therefore, it is important to develop an efficient and secure surgical technique for TOLF. Herein, we describe a method for laminectomyperformed at the thoracic spine using an ultrasonic osteotome combined with a conventional osteotome. This technique can reduce intraoperative complications. This is a relatively safe and easy-to-learn method that should be recommended for the treatment of TOLF.

Laminectomy for the Removal of Thoracic Ossification of the Ligamentum Flavum TOLF Using Ultrasonic and Conventional Osteotomes

We present a methodology for using an ultrasonic osteotome combined with a conventional osteotome for laminectomy in thoracic ossification of the ligamentum flavum. This is a relatively safe and easy-to-learn method that avoids the perioperative complications associated with the conventional method.

Laminectomie pour l’ablation de l’ossification thoracique du ligamentum flavum (TOLF) à l’aide d’ostéotomes ultrasoniques et conventionnels

Thoracic ossification of the ligamentum flavum (TOLF) is a common cause of progressive thoracic myelopathy. TOLF is typically treated with surgical ...