1. Hayvan Etik Fareler üzerinde uzun süre hipertermi gerçekleştirmeden önce prosedürü Kurumsal Hayvan Bakım ve Kullanım Kurulu tarafından onaylandı emin olun. Bu yazıda ele alınan ve beraberindeki video gösterilen tüm prosedürleri İmmünoloji Ulusal Enstitüsü Kurumsal Hayvan Etik Komitesi tarafından onaylanmıştır. 2. Anlamak Hipertermi Tüm Vücut Hipertermi (WBH) 39.5 tutulur inkübatöre konur önceden ısıtılmış kafeslerde ° C. hayvanlar koyarak teslim edilir Protokol standardizasyon sırasında, bir sıcaklık algılayıcı cerrahi fareler implante edildi ve vücut ısısı farklı deneysel koşullar içinde izlenmiştir. Bu ısıtılmış çevre hayvanlar üzerine yerleştirilmesi, bunların sıcaklık yavaş yavaş 37 arttığı görülmektedir ° C ila 42 ° C ile 100 dakika (Şekil 1) bir süre boyunca. Thiç vücut sıcaklığını dış ortam sıcaklığı daha yüksek ateş gibi sıcaklık &quot;başak&quot; sonuçlanan e merak uyandırıcı. Vücut sıcaklığı 40 ° C aştığı zaman hayvan stres ve burun çevresinde ıslatma gibi dehidratasyon gösterecektir. 40 ötesinde vücut sıcaklığı artırın ° C, ıslatma ve tersinmez stres hatta ısı stresi nedeniyle hayvanların ölümüne yol açacaktır. Yani hipertermi sırasında sıcaklık yükselmelerini önleyen önemlidir. Yüksek sıcaklık başak Bu fenomen, hipertermi maruz kalma sırasında hayvan fiziksel hiperaktivite ile ilişkilidir. Benzer sıcaklık ani de diğer gruplar tarafından gözlendi ve çevre 10 yüksek sıcaklıklara sahip tanıtılıyor üzerine hayvanın fiziksel hiperaktivite ile ilişkili bulunmuştur. Bizim protokol, farelerin hipertermi maruziyeti sırasında hareketsiz kalıyor. GibiŞekil 2 de gösterildiği gibi, hayvan immobilizasyonu sıcaklık sivri önler. Bu protokol göstermek için 8-12 haftalık BALB / Cj fareler kullanılmıştır. Protokol belli bir tip / farelerin zorlanma ile sınırlı değildir. 3. Denemeye Hazırlanma 3.1 İnkübatör Bunlar iyi bir sıcaklık kontrol olduğu gibi bu deney, normal bir CO2 kuluçka makinesi içinde gerçekleştirilebilir. CO 2 gaz beslemesini kesin ve CO 2 inkübatör su tutma tepsisini çıkarın. Kuluçka Bazı modellerde bu CO 2 cihazı kapatmak mümkündür. Bir fare kafes rahatlıkla içine yerleştirilmiş olabilir böylece kuluçka rafları. 39.5 ° C&#39;de kuluçka sıcaklığı ayarlayın 3.2 Kafesleri Tipik olarak, hipertermi, deney bir seferde 4 fareler üzerinde gerçekleştirilmiştir. İki kafesleri gereklidir; ilgiliE inkübatör içine yerleştirilir ve diğer oda sıcaklığında dışına yerleştirilir. Yatak yamalar kafes içinde olmayan tekdüze sıcaklığı sonuçlanan zaman ıslanması gibi kafes yatakları, kaçınılmalıdır. Dışında kafes gıda pelet ve su şişesi olmalıdır. 3.3 Fare immobilizasyon tüpler Hipertermi farelerin ilk 4 döngüleri sırasında delikli borular immobilize edilir. Bu sabitleme tüpler gövdesi ve bu tür tüplerin dört gerekli olacaktır fazla delme 3 mm delik ile 50 ml &#39;santrifüj&#39; borudan yapılabilir. 50ml tüp elverişli 8 haftadan daha eski olan BALB / Cj fareler hareketsiz olabilir. Uygun boyutlarda Tüpler farelerin bazı alışılmadık farklı suşu için ya çok genç ya da 50 haftadan daha eski olan fareler için gerekli olabilir. Sıra immobilizasyon tüpleri ve inkübatör içinde bir kafes 39.5 ° C&#39;de muhafaza Böylece en az 2 saat bekledikten sonra, kafes ve immobilization tüpler kuluçka sıcaklığına ulaşmak. 4. Hipertermi ilk çevrimi Fare hipertermi pozlama sırasında dehidratasyon karşılaşabilirsiniz. Bu nedenle deney başlamadan önce, her fare için 0.5 ml serum fizyolojik intraperitoneal olarak yönetmek. 1 ml şırınga ile 26 G iğne kullanın. Çabuk yere, her immobilizasyon tüpler bir hayvan kuvözde kafes çıkarmak ve inkübatör kafese geri yerleştirin. Zaman kaydedin. Bu kuvöz aynı yerde her zaman kafes tutmak esastır. 90 dakika sonra, kuluçka makinesi dan kafes çıkar. Immobilizasyon tüplerinden hayvanlar serbest bırakın ve yiyecek ve su kolay erişim ile normal ısıda kafese koyun. Inkübatöre geri boş immobilizasyon tüpleri yerleştirin. 15 dakika sonra, hipertermi ikinci çevrimi 0 sunarak başladı.Her fare için 5 ml serum fizyolojik intraperitoneal olarak verildi. Daha sonra 50 ml delikli immobilizasyon tüpler içinde immobilize ve inkübatör içine yerleştirilir. 5. Müteakip Hipertermi Çevrimleri İlk 4 kür için, hipertermi pozlama sırasında ve ilk iki döngüleri için fare hareketsiz esastır, salin enjeksiyonu hipertermi dönemi başlamadan önce verilir. Geri kalanı tarafından takip hipertermi bu döngüleri tamamlandıktan sonra, fareler ACCLIMATIZE almak ve hipertermi pozlama sırasında hiperaktif olmazlar. Fareler kolayca dinlenme zaman yiyecek ve su çekmek ise salin intraperitoneal gerekli olmayabilir. 6. Tüm Cycles Tamamlanan sonra, Son Adım Hayvanlar inceleyin etmektir Hipertermi döngüleri sırasında herhangi bir ölüm olmamalıdır. Her hayvan ilk olarak çok dikkatle takip edilmesi gereken bir sahiptirbirkaç döngüleri. Hızla burun etrafında herhangi bir ıslatma arayın. Bu belirti dehidratasyon bir imzadır. Bu semptomlar ile Fareler hemen oda sıcaklığında 0.5 ml serum fizyolojik ip ve kalan verilmelidir. Immobilize farelerin çoğu herhangi bir ek bakım gerektirmez ve başarıyla stres hipertermi taşıyacak. Gerekli döngüleri tamamlandıktan sonra, hayvanlar zayıf ve susuz görünebilir ama onlar hızla birkaç saat içinde iyileşir. <img src="/files/ftp_upload/3801/3801fig1.jpg" alt="Şekil 1" /> Şekil 1. Kafes 39.5 ° C&#39;de bir inkübatöre yerleştirildi Sıçanların vücut çekirdek sıcaklığı değişimi 90 dakika sonra kafes inkübatörden çıkarılmıştır ve 25 ° C&#39;de yerleştirilmiştir Kırmızı çevreler yüksek sıcaklık sivri göstermektedir. <img src="/files/ftp_upload/3801/3801fig2.jpg" alt="Şekil 2," /> Şekil 2. Merkez vücut sıcaklıkları Değişimihipertermi döngüleri sırasında hareketsiz Mice yeniden. Hayvan hareketsizleştirici sonra, yüksek sıcaklıklarda sivri kafes 39.5 ° C&#39;de yerleştirildiği zaman gözlemek değildi

<ol>
	<li>Braude, A. I., Beck, J., Zalesky, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Febrile+response+to+bacterial+pyrogens+in+leukemia.">Febrile response to bacterial pyrogens in leukemia.</a> Blood. 16, 1279 (1960).</li><li>Dovern, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperthermic+intraperitoneal+chemotherapy+added+to+the+treatment+of+ovarian+cancer.+A+review+of+achieved+results+and+complications.">Hyperthermic intraperitoneal chemotherapy added to the treatment of ovarian cancer. A review of achieved results and complications.</a> Eur. J. Gynaecol. Oncol. 31, 256 (2010).</li><li>Gaworek, J., Mayer, C. 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Cancer Res. 9, 2079 ( ).</li><li>Jia, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+B16+murine+melanoma+metastasis+and+enhancement+of+immunity+by+fever-range+whole+body+hyperthermia.">Inhibition of B16 murine melanoma metastasis and enhancement of immunity by fever-range whole body hyperthermia.</a> Int. J. Hyperthermia. 27, 275 (2011).</li><li>Maluta, S., Dall'oglio, S., Nadalini, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+for+intermediate+and+high-risk+prostate+cancer:+controversial+issues+and+the+role+of+hyperthermia.">Treatment for intermediate and high-risk prostate cancer: controversial issues and the role of hyperthermia.</a> Int. J. Hyperthermia. 26, 765 (2010).</li><li>Moyer, H. R., Delman, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+hyperthermia+in+optimizing+tumor+response+to+regional+therapy.">The role of hyperthermia in optimizing tumor response to regional therapy.</a> Int. J. Hyperthermia. 24, 251 (2008).</li><li>Muthana, M., Multhoff, G., Pockley, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumour+infiltrating+host+cells+and+their+significance+for+hyperthermia.">Tumour infiltrating host cells and their significance for hyperthermia.</a> Int. J. Hyperthermia. 26, 247 (2010).</li><li>Ostberg, J. R., Repasky, E. 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Biz ifşa etmek başka bir şey var.

<html> Bildiğim kadarıyla geri 5.000 M.Ö., Mısırlı doktorlar ısı ile tümör tedavi. Yunanlılar, bazı tıbbi tedaviler ısı değeri tanıdı. Musa&#39;ya söz Kaplıcalar (Tekvin 36:24) Kanunu en eski metinler terapötik vücut ısısını yükseltmek için. Önceki teknikler hipertermi 1 indüksiyonu için ateş veya sıcak su banyoları indüksiyonu için pirojenik mikrobiyal ekstrelerinin kullanımı ya istihdam. Ancak bu yaklaşımların hiçbiri kolayca izlenebilir ...

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protocol for long duration whole body hyperthermia in mice

Hipertermi, normal vücut çekirdek yukarıda sıcaklık artışı belirlemek için kullanılan genel bir terimdir. Genellikle ateş sırasında gözlenen artan iç vücut sıcaklığı tanımlamak için kullanılır. Bir yardımcı madde olarak hipertermi kullanımı kanser biyoloji alanında tümör gerilemesi için umut verici bir prosedür olarak ortaya çıkmıştır. Bu amaç için, en önemli gereklilik güvenilir ve düzgün ısıtma protokolleri olmasıdır. Biz farelerde hipertermi (tüm vücut) için bir protokol geliştirmiştir. Bu protokolde, hayvanların 15 dk bir dinlenme dönemi izler 90 dakika hipertermi döngülerine maruz kalmaktadır. Bu dönemde fareler yiyecek ve su kolay erişim vardır. İlk bir kaç hipertermi maruz kalma döngüleri sırasında fareler içinde Yüksek vücut sıcaklığı sivri hayvan hareketsizleştirici tarafından önlenir. Ayrıca, normal salin dehidratasyon etkilerini en aza indirmek için ilk birkaç döngüler halinde uygulanır. Bu protokol 12-24 hr farelerde koşulları gibi ateş simüle edebilirsiniz. Bizprotokolü göstermek için 8-12 haftalık BALB / Cj dişi fareler kullanıldı.

Watch this Scientific Journal Video about Protocol for Long Duration Whole Body Hyperthermia in Mice at JoVE.com

Protocol for Long Duration Whole Body Hyperthermia in Mice

Bu kağıt kadar 12-24 hr koşulları gibi ateş uyarabilir farelerde tüm vücut ısısı için bir protokol açıklar.

Farelerde Uzun Süre Whole Body Hipertermi için protokol

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body ...

protocol-for-long-duration-whole-body-hyperthermia-in-mice

Research

JoVE Journal

Medicine

11.5K Görüntüleme.  National Institute of Immunology. Bu kağıt kadar 12-24 hr koşulları gibi ateş uyarabilir farelerde tüm vücut ısısı için bir protokol açıklar.

Video: Protocol for Long Duration Whole Body Hyperthermia in Mice

Parabiosis in Mice: A Detailed Protocol

Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of microvasculature at the site of inflammation. Parabiotic partners share their circulating antigens and thus are free of adverse immune reaction. First described by Paul Bert in 18641, the parabiosis surgery was refined by Bunster and Meyer in 1933 to improve animal survival2. In the current protocol, two mice are surgically joined following a modification of the Bunster and Meyer technique. Animals are connected through the elbow and knee joints followed by attachment of the skin allowing firm support that prevents strain on the sutured skin. Herein, we describe in detail the parabiotic joining of a ubiquitous GFP expressing mouse to a wild type (WT) mouse. Two weeks after the procedure, the pair is separated and GFP positive cells can be detected by flow cytometric analysis in the blood circulation of the WT mouse. The blood chimerism allows one to examine the contribution of the circulating cells from one animal in the other.

Parabiotic joining of two organisms leads to the development of a shared circulatory system. In this protocol, we describe the surgical steps to form a parabiotic connection between a wild-type mouse and a constitutive GFP-expressing mouse.

Farelerde parabiyoz: Detaylı Bir Protokol

Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of ...

Tıp

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

Farelerde Taksanlar Farmakokinetik ve Farmakodinamik Analiz Karotis Arter İnfüzyonlar

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Long-term Blood Pressure Measurement in Freely Moving Mice Using Telemetry

During the development of new vasoactive agents, arterial blood pressure monitoring is crucial for evaluating the efficacy of the new proposed drugs. Indeed, research focusing on the discovery of new potential therapeutic targets using genetically altered mice requires a reliable, long-term assessment of the systemic arterial pressure variation. Currently, the gold standard for obtaining long-term measurements of blood pressure in ambulatory mice uses implantable radio-transmitters, which require&#160;artery cannulation. This technique eliminates the need for tethering, restraining, or anesthetizing the animals which introduce stress and artifacts during data sampling. However, arterial blood pressure monitoring in mice via catheterization can be rather challenging due to the small size of the arteries. Here we present a step-by-step guide to illustrate the crucial key passages for a successful subcutaneous implantation of radio-transmitters and carotid artery cannulation in mice. We also include examples of long-term blood pressure activity taken from freely moving mice after a period of post-surgery recovery. Following this procedure will allow reliable direct blood pressure recordings from multiple animals simultaneously.

The goal of this protocol is to assess systemic blood pressure in conscious freely moving mice using implantable radio-telemetry devices.

Telemetri kullanarak Serbestçe Hareketli Fare Uzun süreli Tansiyon Ölçümü

During the development of new vasoactive agents, arterial blood pressure monitoring is crucial for evaluating the efficacy of the new proposed drugs. ...

Body Composition and Metabolic Caging Analysis in High Fat Fed Mice

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate utilization, and behaviors such as food intake and physical activity can provide important information regarding the underlying mechanisms of disease. Given the importance of body composition and metabolism to the development of obesity and its subsequent sequelae, it is necessary to make accurate measures of these parameters in the pre-clinical research setting. Advances in technology over the past few decades have made it possible to derive these measures in rodent models in a non-invasive and longitudinal fashion. Consequently, these metabolic measures have proven useful when assessing the response of genetic manipulations (for example knockout or transgenic mice, viral knock-down or overexpression of genes), experimental drug/compound screening and dietary, behavioral or physical activity interventions. Herein, we describe the protocols used to measure body composition and metabolic parameters using an animal monitoring system in chow-fed and high fat diet-fed mice.

This protocol describes the use of a body composition analyzer and metabolic animal monitoring system to characterize body composition and metabolic parameters in mice. An obesity model induced by high-fat feeding is used as an example for the application of these techniques.

Vücut kompozisyonu ve metabolik Caging Analizi yüksek yağ Fed farelerde

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate ...

Protocol to Create Chronic Wounds in Diabetic Mice

Chronic wounds develop as a result of defective regulation in one or more complex cellular and molecular processes involved in proper healing. They impact ~6.5M people and cost ~$40B/year in the US alone. Although a significant effort has been invested in understanding how chronic wounds develop in humans, fundamental questions remain unanswered. Recently, we developed a novel mouse model for diabetic chronic wounds that have many characteristics of human chronic wounds. Using db/db-/- mice, we can generate chronic wounds by inducing high levels of oxidative stress (OS) in the wound tissue immediately after wounding, using a one-time treatment with inhibitors specific to the antioxidant enzymes catalase and glutathione peroxidase. These wounds have high levels of OS, develop biofilm naturally, become fully chronic within 20 days after treatment and can remain open more for more than 60 days. This novel model has many features of diabetic chronic wounds in humans and therefore can contribute significantly to advancing fundamental understanding of how wounds become chronic. This is a major breakthrough because chronic wounds in humans cause significant pain and distress to patients and result in amputation if unresolved. Moreover, these wounds are very expensive and time-consuming to treat, and lead to significant loss of personal income to patients. Advancements in this field of study through the use of our chronic wound model can significantly improve health care for millions who suffer under this debilitating condition. In this protocol, we describe in great detail the procedure to cause acute wounds to become chronic, which has not been done before.

Chronic wounds are developed from acute wounds on a diabetic mouse model by inducing high levels of oxidative stress after a full-thickness cutaneous wound. The wound is treated with inhibitors for catalase and glutathione peroxidase, resulting in impaired healing and biofilm development by bacteria present in the skin microbiome.

Diyabetik Farelerde Kronik Yaralar Oluşturma Protokolü

Chronic wounds develop as a result of defective regulation in one or more complex cellular and molecular processes involved in proper healing. They impact ...

Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children

Coins are the most common ingested metallic foreign bodies among children. The goal of this protocol is to assess the accuracy and feasibility of using a handheld metal detector to detect ingested metallic foreign bodies in children. We propose that by introducing handheld metal detector screening early in the triage process of children with high suspicion of metallic foreign body ingestion, the number of radiographs being ordered to localize the metallic foreign body can be reduced in this radio-sensitive population. The study protocol requires the screening of the participants for history of foreign body ingestion and exclusion of patients with respiratory distress or metallic implants. The patient changes to hospital gown and items that could contain metal like eyeglasses, earrings, pendants, and ornaments are removed. The patient is positioned in the center of the room away from other metallic interferences. The working status of the handheld metal detector is first confirmed by eliciting a positive audio-visual signal. Then the screening is done in an erect position with head in extension to expose the neck, from the level of the chin to the level of the hip joint, to cover the anatomical areas from neck to pelvis in a zig-zag manner both anteriorly and posteriorly. A positive audio-visual signal is carefully noted during the scanning for the presence of metallic foreign body. Relevant radiographs are ordered as per the area detected on the metal detector screening. The handheld metal detector was able to precisely identify all the coins among the ingested metallic foreign bodies in our study. The handheld metal detector could not consistently detect non-coin metallic foreign bodies. This protocol demonstrates the accuracy of handheld metal detector in the identification and localization of coins and coin like metallic foreign bodies.

Here, we present a protocol to use the handheld metal detector, to screen for the presence of ingested metallic foreign bodies in children who present to the pediatric emergency medicine department with a history of foreign body ingestion.

Metalik yabancı cisim yeme çocuklar için el Metal detektörü tarama

Coins are the most common ingested metallic foreign bodies among children. The goal of this protocol is to assess the accuracy and feasibility of using a ...

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to expend energy has made it a potential target for novel therapies to ameliorate obesity and associated metabolic disorders in humans. Though this tissue has been well studied in small animals, BAT&#39;s thermogenic capacity in humans remains largely unknown due to the difficulties of measuring its volume, activity, and distribution. Identifying and quantifying active human BAT is commonly performed using 18F-Fluorodeoxyglucose (18F-FDG) positron emission tomography and computed tomography (PET/CT) scans following cold-exposure or pharmacological activation. Here we describe a detailed image-analysis approach to quantify total-body human BAT from 18F-FDG PET/CT scans using an open-source software. We demonstrate the drawing of user-specified regions of interest to identify metabolically active adipose tissue while avoiding common non-BAT tissues, to measure BAT volume and activity, and to further characterize its anatomical distribution. Although this rigorous approach is time-consuming, we believe it will ultimately provide a foundation to develop future automated BAT quantification algorithms.

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

Using free, open-source software, we have developed an analytical approach to quantify total and regional brown adipose tissue (BAT) volume and metabolic activity of BAT using 18F-FDG PET/CT.

18F-FDG PET/BT kullanarak yağ dokusu kahverengi tüm vücut ve bölgesel Quantification etkin insan

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to ...

A Whole Body Dosimetry Protocol for Peptide-Receptor Radionuclide Therapy (PRRT): 2D Planar Image and Hybrid 2D+3D SPECT/CT Image Methods

Peptide-receptor-radionuclide-therapy (PPRT) is a targeted therapy that combines a short-range energy radionuclide with a substrate with high specificity for cancer cell receptors. After injection, the radiotracer is distributed throughout the entire body, with a higher uptake in tissues where targeted receptors are overexpressed. The use of beta/gamma radionuclide emitters enables therapy imaging (beta-emission) and post-therapy imaging (gamma-emission) to be performed at the same time. Post-treatment sequential images permit absorbed dose calculation based on local uptake and wash-in/wash-out kinetics. We implemented a hybrid method that combines information derived from both 2D and 3D images. Serial whole-body images and blood samples are acquired to estimate the absorbed dose to different organs at risk and to lesions disseminated throughout the body. A single 3D-SPECT/CT image, limited to the abdominal region, overcomes projection overlap on planar images of different structures such as the intestines and kidneys. The hybrid 2D+3D-SPECT/CT method combines the effective half-life information derived from 2D planar images with the local uptake distribution derived from 3D images. We implemented this methodology to estimate the absorbed dose for patients undergoing PRRT with 177Lu-PSMA-617. The methodology could, however, be implemented with other beta-gamma radiotracers. To date, 10 patients have been enrolled into the dosimetry study with 177Lu-PSMA-617 combined with drug protectors for kidneys and salivary glands (mannitol and glutamate tablets, respectively). The median ratio between kidney uptake at 24 h evaluated on planar images and 3D-SPECT/CT is 0.45 (range:0.32-1.23). The comparison between hybrid and full 3D approach has been tested on one patient, resulting in a 1.6% underestimation with respect to full 3D (2D: 0.829 mGy/MBq, hybrid: 0.315 mGy/MBq, 3D: 0.320 mGy/MBq). Treatment safety has been confirmed, with a mean absorbed dose of 0.73 mGy/MBq (range:0.26-1.07) for kidneys, 0.56 mGy/MBq (0.33-2.63) for the parotid glands and 0.63 mGy/MBq (0.23-1.20) for submandibular glands, values in accordance with previously published data.

A Whole Body Dosimetry Protocol for Peptide-Receptor Radionuclide Therapy PRRT: 2D Planar Image and Hybrid 2D+3D SPECT/CT Image Methods

This method estimates the absorbed dose of different structures for peptide-receptor-radionuclide-therapy (PRRT) with the possibility of avoiding organ overlap on 2D-projections. Serial whole-body planar images permit estimation of mean absorbed doses along the whole body, while the hybrid approach, combining planar images and 3D-SPECT/CT image, overcomes the limitations of structure overlapping.

Peptit-Reseptör Radyonüklid Tedavisi (PRRT) için Tüm Vücut Dozimetri Protokolü : 2D Düzlemsel Görüntü ve Hibrid 2D+3D SPECT/BT Görüntü Yöntemleri

Peptide-receptor-radionuclide-therapy (PPRT) is a targeted therapy that combines a short-range energy radionuclide with a substrate with high specificity ...

In Vitro and In Vivo Delivery of Magnetic Nanoparticle Hyperthermia Using a Custom-Built Delivery System

Hyperthermia has long been used in the treatment of cancer. Techniques have varied from the intra-tumoral insertion of hot iron rods, to systemically delivered tumor antibody-targeted magnetic nanoparticles, at temperatures from 39 &#730;C (fever-level) to 1,000 &#730;C (electrocautery) and treatment times from seconds to hours. The temperature-time relationship (thermal dose) dictates the effect with high thermal doses resulting in the tissue ablation and lower thermal doses resulting in sublethal effects such as increased blood flow, accumulation of drugs and immune stimulation. One of the most promising current medical therapies is magnetic nanoparticle hyperthermia&#160; (mNPH). This technique involves activating magnetic nanoparticles, that can be delivered systemically or intratumorally, with a non-invasive, non-toxic alternating magnetic field. The size, construct and association of the magnetic nanoparticles and the frequency and field strength of the magnetic field are major heating determinants. We have developed sophisticated instrumentation and techniques for delivering reproducible magnetic nanoparticle hyperthermia in large and small animal models and cultured cells. This approach, using continuous, real time temperature monitoring in multiple locations, allows for the delivery of well-defined thermal doses to the target tissue (tumor) or cells while limiting non-target tissue heating. Precise control and monitoring of temperature, in multiple sites, and use of the industry standard algorithm (cumulative equivalent minutes at 43 &#730;C /CEM43), allows for an accurate determination and quantification of thermal dose. Our system, which allows for a wide variety of temperatures, thermal doses, and biological effects, was developed through a combination of commercial acquisitions and inhouse engineering and biology developments. This system has been optimized in a manner that allows for the rapid conversion between ex vivo, in vitro, and in vivo techniques. The goal of this protocol is to demonstrate how to design, develop and implement an effective technique and system for delivering reproducible and accurate magnetic nanoparticle therapy (mNP) hyperthermia.

This protocol presents techniques and methodology necessary for the accurate delivery of magnetic nanoparticle hyperthermia using a sophisticated delivery and monitoring system.

Özel Yapım Bir Dağıtım Sistemi Kullanarak Manyetik Nanopartikül Hipertermisinin In Vitro ve In Vivo Teslimatı

Hyperthermia has long been used in the treatment of cancer. Techniques have varied from the intra-tumoral insertion of hot iron rods, to systemically ...

Assessing Whole-Body Lipid-Handling Capacity in Mice

Assessing lipid metabolism is a cornerstone of evaluating metabolic function, and it is considered essential for in vivo metabolism studies. Lipids are a class of many different molecules with many pathways involved in their synthesis and metabolism. A starting point for evaluating lipid hemostasis for nutrition and obesity research is needed. This paper describes three easy and accessible methods that require little expertise or practice to master, and that can be adapted by most labs to screen for lipid-metabolism abnormalities in mice. These methods are (1) measuring several fasting serum lipid molecules using commercial kits (2) assaying for dietary lipid-handling capability through an oral intralipid tolerance test, and (3) evaluating the response to a pharmaceutical compound, CL 316,243, in mice. Together, these methods will provide a high-level overview of lipid handling capability in mice.

This paper provides three easy and accessible assays for assessing lipid metabolism in mice.

Farelerde Tüm Vücut Lipid Taşıma Kapasitesinin Değerlendirilmesi

Assessing lipid metabolism is a cornerstone of evaluating metabolic function, and it is considered essential for in vivo metabolism studies. Lipids are a ...