저자는 박사 리 카이 핀, 머리 & 수석 컨설턴트, 어린이 비상, KK 여자와 아동 병원, 100 부 킷 Timah도로, 싱가포르 비디오 문서 생산 관리 지원에 대 한 감사합니다. 문서는 "휴대용 금속 탐지기에 대 한 금속 외국 몸 섭취에 소아 응급";에서 허가로 재현 8 월 2018: 85 (8): 618-624; Hazwani Binte Hamzah, 철 야 제임스, Suraj Manickam, Sashikumar Ganapathy: 인도 저널 소아과 저작권.

프로토콜 인간의 기관 연구 윤리 위원회의 지침을 따릅니다. 중앙 기관 검토 위원회는 연구 윤리 승인 제공.
1입니다. 참가자 심사
<ol><li>이물질 섭취의 역사와 함께 소아 응급 부서에 선물 하는 환자를 모집 합니다.</li>
	<li>호흡 곤란에는 속도 제조 업체와 같은 장치를 이식 했다 환자를 제외 합니다.</li>
</ol>2. 환자 준비
<ol><li>환자는 병원 가운에서 드레스.</li>
	<li>옷, 안경, 귀걸이, 펜 던 트, 및 HHMD 심사 간섭을 유발 하지 않도록 심사 전에 장식 금속 개체를 포함할 수 있는 모든 항목을 제거 합니다.</li>
	<li>다른 잠재적인 금속 방해에서 공간의 중심에 환자를 배치 합니다.</li>
</ol>3. 심사 관 및 보호자 준비
<ol><li>보석, 시계 등 모든 금속 액세서리를 제거 합니다.</li>
	<li>보석, 시계 등 모든 금속 액세서리를 제거 하는 간병인을 요청 합니다.</li>
</ol>4. 금속 탐지기 검사
<ol><li>HHMD (그림 1)를 사용 하 여 심사를 실시 하.</li>
	<li>금속 조각에 쳐서 긍정적인 시청각 신호를 도출 하 여는 HHMD의 작동 상태를 확인 합니다.</li>
	<li>HHMD 더 큰 아이 들을 위한 서 있는 위치에서 검색을 수행 합니다.</li>
	<li>HHMD 교사 잡아 환자를 적절 하 게 함으로써 유아를 위한 직 립 위치에 검색을 수행 합니다.</li>
	<li>그리고 본문에서 환자의 손을 잡으십시오.</li>
	<li>위치는 환자의 머리 목 노출에 대 한 확장입니다.</li>
	<li>고관절 골반을 목에서 해 부 영역을 커버 하의 수준에는 턱의 수준에서 스캔.</li>
	<li>지그재그 방식으로 모든 지역 검색 되도록 몸 전체 스캔을 수행 합니다.</li>
	<li>스캔 모두 anteriorly과 뒤로 환자의 두께 고려 사항으로 촬영 해야 합니다.</li>
	<li>신중 하 게 MFBs의 존재에 대 한 검색 하는 동안 긍정적인 시청각 신호 note.</li>
</ol>5. 수행 관련 방사선 사진
<ol><li>MFBs의 HHMD 검사에 의해 감지 영역에 의하여 목, 가슴, 또는 복 부의 관련 방사선 사진 주문.</li>
	<li>이물질 섭취는 높은 MFB HHMD 심사 및 의혹의 임상 색인 하는 동안 검색 되지 않습니다 경우 모든 x 선 조회 (목, 가슴, 복 부) 진행.</li>
</ol>6. 테스트 결과 기록
<ol><li>기록 일지 (그림 2) 특히 테스트 결과 워크플로 (그림 1)에 따라.</li>
	<li>Palpating는 다음과 같은 저명한 앙상한 랜드마크-sternal 노치, xiphisternum, 음모 관절, 그리고 양측 장 골도 머리에 의해 시체에, 기본 뼈 랜드마크 긍정적인 시청각 HHMD 신호 상관.</li>
	<li>X 선 검사를 진행 하기 전에 항해 일지의 해 부 참조 개요에 랜드마크를 기록 합니다.</li>
</ol>

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저자는 공개 없다.

금속 어린이 몸 섭취 일반적인 문제 이며 환자의 대다수에서 이것은 외국 관리 보수적7,16. 단일 또는 반복 검사는 섭취 금속 이물질을 지역화 하는 동안 원치 않는 방사선 위험 라디오-민감한 소아 인구를 노출할 수 있습니다. 어린이, 감도와 보다 96%와 801,7,17,,<sup cla...

이물질 섭취 시음 하 고 그들에 게1,2를 삼 키는 의해 개체를 탐구 하는 유아와 어린이의 자연 호기심 많은 성격 때문에 소아 응급실을 제시 하는 아이 들 사이에서 광범위 한 문제입니다. 이전 연구는 금속 물체 환자3,4의 그들의 일대에 모든 이물질 ingestions의 85%를 구성 했다 보고 있다. 이 프로토콜의 목표는 정확성과 어린이 섭취 금속 외국 시체를 감지 하는 휴대용 금속 탐지기를 사용 하 여의 타당성을 평가 하는입니다. 우리가 제안 하는 휴대용 금속 탐지기 금속 이물질 섭취의 높은 의혹으로 어린이의 심사 과정에서 초기 프로토콜 검사를 도입 하 여 방사선 금속 이물질을 지역화 하도록 주문 되 고 수가 될 수 있습니다. 이 라디오-민감한 인구 감소.
휴대용 금속 탐지기 (HHMD) 저렴 하 고 쉽게 접근할 수 있는 외래 섭취 금속 외국 기관의 관리를 신속 하 게 하는 것을 도울 수 있는 이다. 금속 탐지기는 수신기와 송신기의 주변3금속 개체의 존재에 기인한 전자기 분야에서 변화를 감지 하는 매우 낮은 주파수 장치입니다. 금속 개체 필드 내에 있으면 전자기장 방해 하 고 센서는 시청각 경보 장치4에 설정 됩니다. 금속 탐지기 검사 2 분5미만에 철저 하 게 수행할 수 있습니다. 따라서, HHMD 심사 지형 지역화 금속 이물질 섭취의 높은 의혹으로 어린이의 심사 프로세스의 초기에 도입 될 수 있습니다.
동전은 가장 일반적인 삼 켜 금속 외국 기관1,4,,67. 날카로운 모서리와 일반 비 독성 자연의 부족, 동전 ingestions의 대다수는 보수적으로 관리 됩니다. 일반적으로 섭취 하는 물질의 다른 종류는 자석, 단추 배터리, 작은 장난감, 땅콩, 보석, 단추, 뼈, 씨앗, 그리고 팝콘8,9포함 됩니다. 외국 기관의 impaction 3 사이트, 즉 상부 식도 괄약근, 대동맥 아치, 그리고 더 낮은 식도 괄약근10의 수준에 중간 식도에서 식도에 가장 일반적으로 발생합니다. 식도에 있는 금속 외국 기관 (MFBs) 환자11,12 의 40%에서 무 증상 수 있으며 그것의 장기간된 impaction 식도 천공, mediastinitis, 같은 다양 한 합병증으로 이어질 수 있습니다. tracheoesophageal 누, tracheal 협 착 증, 및 aorto-식도 누 관1,13,,1415. 이물질 impaction의 다른 위치 등 pylorus, 십이지 장, ileo caecal 접속점10.
목, 가슴 및 복 부를 진단 하는 검사를 수행 의심 이물질 섭취 비상 부서5에 대폭적인 연습 이다. 섭취 한 외국의 모든 종류, 중 10%는 라디오 불투명10. 방사능 테스트 시간이 고 어린이 방사선 노출의 위험이 있다. 섭취 한 금속 물체의 정확한 지형 지역화는 매우 중요 한 치료 가이드를 합니다. HHMD 초기 심사 과정에서에서 심사의 도입 지형학 섭취 금속 이물질을 지역화 하 여 방사선의 외부 주문 완화 도울 수 있습니다.
이 연구 체계적인 지형 패션에 MFBs를 지역화 하는 HHMD의 정확도 평가 하 고 효율성과 지역 소아과 인구에 있는 HHMD의 타당성을 결정. 우리는 또한 얼마나 잘이 방사선 검사 도구 무료 환자와 부모에 의해 받아들여졌다 조사 했다. 5 월과 7 월 2016 년 사이 이물질 섭취의 역사와 함께 소아 응급 부서에 제시 모든 연속 환자는 연구에서 포함 되었다. 동의 부모 또는 법적 보호자 로부터 얻은 했다. 추정 된 이물질 섭취 환자 체계적으로 HHMD를 사용 하 여 검사 되었다. 식별에 대 한 치료의 골드 표준 및 섭취 금속 외국 기관의 지역화 일반 검사 자 궁 경부 지역, 흉부, 그리고 복 부의 수행 이다. 연구의 수행을 촉진 하기 위하여 표준화 된 워크플로 (그림 1)을 만들었습니다. 특히 워크플로 기반으로 만든 로그도 서 스프레드시트 (그림 2) 이후 변하게 했다 초기 데이터 수집에 사용 되었다. 결과 섹션에 데이터 백분율에 의해 다음의 경우의 수로 묘사 된다.

<table>
	<tbody>
		<tr>
			<td>Hand held metal detector</td>
			<td>Garrett Super Scanner V</td>
			<td>NA</td>
			<td>NA</td>
		</tr>
	</tbody>
</table>

handheld metal detector screening for metallic foreign body ingestion in children

동전은 아이 들 중 가장 일반적인 섭취 금속 외국인 시체. 이 프로토콜의 목표는 정확성과 어린이 섭취 금속 외국 시체를 감지 하는 휴대용 금속 탐지기를 사용 하 여의 타당성을 평가 하는입니다. 우리는 금속 이물질 섭취의 높은 의혹으로 어린이의 심사 프로세스의 초기에 휴대용 금속 탐지기 검사를 도입 하 여 금속 이물질을 지역화 하도록 주문 되는 방사선의 수 감소 될 수 있다이 제안 라디오-민감한 인구입니다. 연구 프로토콜 이물질 섭취의 역사와 호흡 곤란 환자의 배제에 대 한 참가자의 심사 또는 금속 임 플 란 트. 환자 병원 가운과 안경, 귀걸이, 펜 던 트, 같은 금속을 포함할 수 있는 항목을 변경 하 고 장신구 제거 됩니다. 환자는 다른 금속 방해에서 방 중앙에 배치 됩니다. 휴대용 금속 탐지기의 작동 상태는 먼저 긍정적인 시청각 신호를 도출 하 여 확인 됩니다. 다음 심사 목 anteriorly과 뒤로 지그재그 방식으로 골반을 목에서 해 부 영역을 커버 하는 엉덩이 관절의 수준에 턱의 수준에서 노출에 대 한 확장에 머리는 직 립 위치에 이루어집니다. 긍정적인 시청각 신호 금속 이물질의 존재에 대 한 검색 하는 동안 신중 하 게 주의 된다. 관련 검사 금속 탐지기 검사에는 지역에 따라 정렬 됩니다. 휴대용 금속 탐지기 정확 하 게 우리의 연구에서 섭취 금속 외국 시체 들 사이 있는 모든 동전을 확인할 수 있었다. 휴대용 금속 탐지기 감지할 수 없습니다 일관 되 게 동전이 아닌 금속 외국인 시체. 이 프로토콜 식별에서 휴대용 금속 탐지기의 정확도 및 동전과 금속 외국 시체 처럼 동전의 지역화를 보여 줍니다.

연구팀은이 관찰 연구에 대 한 36 환자 모집. 이 환자는 이물질 섭취의 역사와 함께 소아 응급 부서에 제시 했다. 이러한 가운데, 28 환자 금속 이물질 ingestions (그림 3) 했다. 나머지 8 환자 물고기 뼈, satay 스틱, 플라스틱 버튼 등 비금속 외국 시체를 섭취 했다. 동전 가장 일반적인 유형의 이물질 섭취 (표 1), 환자의 30.5%를 구성 했다. 다른 ?...

Watch this Scientific Journal Video about Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children at JoVE.com

Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children

여기, 우리 아이 이물질 섭취의 역사와 함께 소아 응급 의학 부서에 제시 들 섭취 금속 외국인 시체의 존재에 대 한 화면을 휴대용 금속 탐지기를 사용 하는 프로토콜을 제시.

아이 들에서 금속 이물질 섭취에 대 한 휴대용 금속 탐지기 검사

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-screening-for-metallic-foreign-body-ingestion

Research

JoVE Journal

Medicine

10.6K 조회수.  KK women's and Children's Hospital. 여기, 우리 아이 이물질 섭취의 역사와 함께 소아 응급 의학 부서에 제시 들 섭취 금속 외국인 시체의 존재에 대 한 화면을 휴대용 금속 탐지기를 사용 하는 프로토콜을 제시.

비디오: Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children

High Content Screening in Neurodegenerative Diseases

The functional annotation of genomes, construction of molecular networks and novel drug target identification, are important challenges that need to be addressed as a matter of great urgency1-4. Multiple complementary 'omics' approaches have provided clues as to the genetic risk factors and pathogenic mechanisms underlying numerous neurodegenerative diseases, but most findings still require functional validation5. For example, a recent genome wide association study for Parkinson's Disease (PD), identified many new loci as risk factors for the disease, but the underlying causative variant(s) or pathogenic mechanism is not known6, 7. As each associated region can contain several genes, the functional evaluation of each of the genes on phenotypes associated with the disease, using traditional cell biology techniques would take too long. 
	There is also a need to understand the molecular networks that link genetic mutations to the phenotypes they cause. It is expected that disease phenotypes are the result of multiple interactions that have been disrupted. Reconstruction of these networks using traditional molecular methods would be time consuming. Moreover, network predictions from independent studies of individual components, the reductionism approach, will probably underestimate the network complexity8. This underestimation could, in part, explain the low success rate of drug approval due to undesirable or toxic side effects. Gaining a network perspective of disease related pathways using HT/HC cellular screening approaches, and identifying key nodes within these pathways, could lead to the identification of targets that are more suited for therapeutic intervention.
	High-throughput screening (HTS) is an ideal methodology to address these issues9-12. but traditional methods were one dimensional whole-well cell assays, that used simplistic readouts for complex biological processes. They were unable to simultaneously quantify the many phenotypes observed in neurodegenerative diseases such as axonal transport deficits or alterations in morphology properties13, 14. This approach could not be used to investigate the dynamic nature of cellular processes or pathogenic events that occur in a subset of cells. To quantify such features one has to move to multi-dimensional phenotypes termed high-content screening (HCS)4, 15-17. HCS is the cell-based quantification of several processes simultaneously, which provides a more detailed representation of the cellular response to various perturbations compared to HTS. 
	HCS has many advantages over HTS18, 19, but conducting a high-throughput (HT)-high-content (HC) screen in neuronal models is problematic due to high cost, environmental variation and human error. In order to detect cellular responses on a 'phenomics' scale using HC imaging one has to reduce variation and error, while increasing sensitivity and reproducibility. 
	Herein we describe a method to accurately and reliably conduct shRNA screens using automated cell culturing20 and HC imaging in neuronal cellular models. We describe how we have used this methodology to identify modulators for one particular protein, DJ1, which when mutated causes autosomal recessive parkinsonism21. 
	Combining the versatility of HC imaging with HT methods, it is possible to accurately quantify a plethora of phenotypes. This could subsequently be utilized to advance our understanding of the genome, the pathways involved in disease pathogenesis as well as identify potential therapeutic targets.

We describe a methodology combining automated cell culturing with high-content imaging to visualize and quantify multiple cellular processes and structures, in a high-throughput manner. Such methods can aid in the further functional annotation of genomes as well as identify disease gene networks and potential drug targets.

Neurodegenerative 질병 높은 콘텐츠 심사

The functional annotation of genomes, construction of molecular networks and novel drug target identification, are important challenges that need to be ...

연구

의학

Eye Tracking Young Children with Autism

The rise of accessible commercial eye-tracking systems has fueled a rapid increase in their use in psychological and psychiatric research. By providing a direct, detailed and objective measure of gaze behavior, eye-tracking has become a valuable tool for examining abnormal perceptual strategies in clinical populations and has been used to identify disorder-specific characteristics1, promote early identification2, and inform treatment3. In particular, investigators of autism spectrum disorders (ASD) have benefited from integrating eye-tracking into their research paradigms4-7. Eye-tracking has largely been used in these studies to reveal mechanisms underlying impaired task performance8 and abnormal brain functioning9, particularly during the processing of social information1,10-11. While older children and adults with ASD comprise the preponderance of research in this area, eye-tracking may be especially useful for studying young children with the disorder as it offers a non-invasive tool for assessing and quantifying early-emerging developmental abnormalities2,12-13. Implementing eye-tracking with young children with ASD, however, is associated with a number of unique challenges, including issues with compliant behavior resulting from specific task demands and disorder-related psychosocial considerations. In this protocol, we detail methodological considerations for optimizing research design, data acquisition and psychometric analysis while eye-tracking young children with ASD. The provided recommendations are also designed to be more broadly applicable for eye-tracking children with other developmental disabilities. By offering guidelines for best practices in these areas based upon lessons derived from our own work, we hope to help other investigators make sound research design and analysis choices while avoiding common pitfalls that can compromise data acquisition while eye-tracking young children with ASD or other developmental difficulties.

Eye tracking has long been used to study gaze patterns in typically-developing individuals, but recent technological advancements have made its use with clinical populations, including autism, more feasible. While eye-tracking young children with autism can offer insight into early symptom manifestations, it involves methodological challenges. Suggestions for best practices are provided.

자폐증과 어린 아이를 추적 눈

The rise of accessible commercial eye-tracking systems has fueled a rapid increase in their use in psychological and psychiatric research. By providing a ...

Protocol for Long Duration Whole Body Hyperthermia in Mice

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 temperature that is observed during fever. The use of hyperthermia as an adjuvant has emerged as a promising procedure for tumor regression in the field of cancer biology. For this purpose, the most important requirement is to have reliable and uniform heating protocols. We have developed a protocol for hyperthermia (whole body) in mice. In this protocol, animals are exposed to cycles of hyperthermia for 90 min followed by a rest period of 15 min. During this period mice have easy access to food and water. High body temperature spikes in the mice during first few hyperthermia exposure cycles are prevented by immobilizing the animal. Additionally, normal saline is administered in first few cycles to minimize the effects of dehydration. This protocol can simulate fever like conditions in mice up to 12-24 hr. We have used 8-12 weeks old BALB/Cj female mice to demonstrate the protocol.

This paper describes a protocol for whole body hyperthermia in mice that can stimulate fever like conditions up to 12-24 hr.

마우스의 긴 기간 온 몸이 고열에 대한 프로토콜

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 ...

Measuring Cardiac Autonomic Nervous System (ANS) Activity in Children

The autonomic nervous system (ANS) controls mainly automatic bodily functions that are engaged in homeostasis, like heart rate, digestion, respiratory rate, salivation, perspiration and renal function. The ANS has two main branches: the sympathetic nervous system, preparing the human body for action in times of danger and stress, and the parasympathetic nervous system, which regulates the resting state of the body.
ANS activity can be measured invasively, for instance by radiotracer techniques or microelectrode recording from superficial nerves, or it can be measured non-invasively by using changes in an organ's response as a proxy for changes in ANS activity, for instance of the sweat glands or the heart. Invasive measurements have the highest validity but are very poorly feasible in large scale samples where non-invasive measures are the preferred approach. Autonomic effects on the heart can be reliably quantified by the recording of the electrocardiogram (ECG) in combination with the impedance cardiogram (ICG), which reflects the changes in thorax impedance in response to respiration and the ejection of blood from the ventricle into the aorta. From the respiration and ECG signals, respiratory sinus arrhythmia can be extracted as a measure of cardiac parasympathetic control. From the ECG and the left ventricular ejection signals, the preejection period can be extracted as a measure of cardiac sympathetic control. ECG and ICG recording is mostly done in laboratory settings. However, having the subjects report to a laboratory greatly reduces ecological validity, is not always doable in large scale epidemiological studies, and can be intimidating for young children. An ambulatory device for ECG and ICG simultaneously resolves these three problems.
Here, we present a study design for a minimally invasive and rapid assessment of cardiac autonomic control in children, using a validated ambulatory device 1-5, the VU University Ambulatory Monitoring System (VU-AMS, Amsterdam, the Netherlands,<a href="http://www.vu-ams.nl" target="_blank"> www.vu-ams.nl</a>).

Measuring Cardiac Autonomic Nervous System ANS Activity in Children

Measurement of autonomic nervous system activity usually confines the researcher and participant to the laboratory, which may provide an intimidating environment to children. The VU University Ambulatory Monitoring System (VU-AMS) device can record cardiac autonomic control in any setting. The VU-AMS proved very amenable to testing in children.

아이들의 측정 심장 자율 신경계 (ANS) 활동

The autonomic nervous system (ANS) controls mainly automatic bodily functions that are engaged in homeostasis, like heart rate, digestion, respiratory ...

Screening for Melanoma Modifiers using a Zebrafish Autochthonous Tumor Model

Genomic studies of human cancers have yielded a wealth of information about genes that are altered in tumors1,2,3. A challenge arising from these studies is that many genes are altered, and it can be difficult to distinguish genetic alterations that drove tumorigenesis from that those arose incidentally during transformation. To draw this distinction it is beneficial to have an assay that can quantitatively measure the effect of an altered gene on tumor initiation and other processes that enable tumors to persist and disseminate. Here we present a rapid means to screen large numbers of candidate melanoma modifiers in zebrafish using an autochthonous tumor model4 that encompasses steps required for melanoma initiation and maintenance. A key reagent in this assay is the miniCoopR vector, which couples a wild-type copy of the mitfa melanocyte specification factor to a Gateway recombination cassette into which candidate melanoma genes can be recombined5. The miniCoopR vector has a mitfa rescuing minigene which contains the promoter, open reading frame and 3'-untranslated region of the wild-type mitfa gene. It allows us to make constructs using full-length open reading frames of candidate melanoma modifiers. These individual clones can then be injected into single cell Tg(mitfa:BRAFV600E);p53(lf);mitfa(lf)zebrafish embryos. The miniCoopR vector gets integrated by Tol2-mediated transgenesis6 and rescues melanocytes. Because they are physically coupled to the mitfa rescuing minigene, candidate genes are expressed in rescued melanocytes, some of which will transform and develop into tumors. The effect of a candidate gene on melanoma initiation and melanoma cell properties can be measured using melanoma-free survival curves, invasion assays, antibody staining and transplantation assays.

A rapid way to screen for melanoma modifiers using a zebrafish autochthonous tumor model is presented. It takes advantage of the miniCoopR vector which allows for expression of candidate melanoma genes in melanocytes. A method to obtain melanoma-free survival curves, an invasion assay, a protocol for antibody staining of scale melanocytes and a melanoma transplantation assay are described.

Zebrafish Autochthonous 종양 모델을 사용하여 흑종 조절을위한 검사

Genomic studies of  human cancers have yielded a wealth of information about genes that are altered  in tumors1,2,3. A challenge arising from these ...

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a novel platform that allows for direct long-term visualization of tissue structure changes intracranially. Imaging at a single cell resolution in a real-time fashion provides supplementary dynamic information beyond that provided by standard end-point histological analysis, which looks solely at 'snap-shot' cross sections of tissue.
Establishing this intravital imaging technique in fluorescent chimeric mice, we are able to image four fluorescent channels simultaneously. By incorporating fluorescently labeled cells, such as GFP+ bone marrow, it is possible to track the fate of these cells studying their long-term migration, integration and differentiation within tissue. Further integration of a secondary reporter cell, such as an mCherry glioma tumor line, allows for characterization of cell:cell interactions. Structural changes in the tissue microenvironment can be highlighted through the addition of intra-vital dyes and antibodies, for example CD31 tagged antibodies and Dextran molecules.
Moreover, we describe the combination of our ICW imaging model with a small animal micro-irradiator that provides stereotactic irradiation, creating a platform through which the dynamic tissue changes that occur following the administration of ionizing irradiation can be assessed.
Current limitations of our model include penetrance of the microscope, which is limited to a depth of up to 900 &mu;m from the sub cortical surface, limiting imaging to the dorsal axis of the brain. The presence of the skull bone makes the ICW a more challenging technical procedure, compared to the more established and utilized chamber models currently used to study mammary tissue and fat pads 5-7. In addition, the ICW provides many challenges when optimizing the imaging.

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We describe a novel in vivo imaging technique that couples fluorescent chimeric mice with intracranial windows and high-resolution 2-photon microscopy. This imaging platform aids studies of dynamic changes in brain tissue and microvasculature, at a single-cell level, following pathological insults and is adaptable to assess intracranial drug delivery and distribution.

소설 고해상도<em&gt; 생체 내</em종양의 성장과 치료학에 두개 내 구조물의 동적 응답을 공부하기&gt; 이미징 기술

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a ...

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone resorption. Osteoclasts can be generated in vitro from monocyte/macrophage precursor cells in the presence of certain cytokines, which promote survival and differentiation. Here, both in vivo and in vitro techniques are demonstrated, which allow scientists to study different cytokine contributions towards osteoclast differentiation, signaling, and activation. The minicircle DNA delivery gene transfer system provides an alternative method to establish an osteoporosis-related model is particularly useful to study the efficacy of various pharmacological inhibitors in vivo. Similarly, in vitro culturing protocols for producing osteoclasts from human precursor cells in the presence of specific cytokines enables scientists to study osteoclastogenesis in human cells for translational applications. Combined, these techniques have the potential to accelerate drug discovery efforts for osteoclast-specific targeted therapeutics, which may benefit millions of osteoporosis and arthritis patients worldwide.

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation of precursor cells into osteoclasts is regulated by cytokines and growth factors. Here, a novel gene transfer technique for differentiation of osteoclasts in vivo and cell culture protocols for differentiating precursor cells into osteoclasts in vitro as a method to study the effects of cytokines on osteoclastogenesis are described.

소설<em&gt; 생체</em&gt; 유전자 전달 기술과<em&gt; 시험관</em근골격계 질환의 뼈 손실의 연구를위한&gt; 셀 기반 분석 실험

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone ...

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a serious problem. Therefore, the acceptance criteria for organ donors have been extended leading to the usage of marginal kidney grafts. These marginal organs tolerate cold storage poorly resulting in increased preservation injury and higher rates of delayed graft function. To overcome the limitations of cold storage, extensive research is focused on alternative normothermic preservation methods.
Ex vivo normothermic organ perfusion is an innovative preservation technique. The first experimental and clinical trials for ex vivo lung, liver, and kidney perfusions demonstrated favorable outcomes.
In addition to the reduction of cold ischemic injury, the method of normothermic kidney storage offers the opportunity for organ assessment and repair. This manuscript provides information about kidney retrieval, organ preservation techniques, and isolated ex vivo normothermic kidney perfusion (NEVKP) in a porcine model. Surgical techniques, set up for the perfusion solution and the circuit, potential assessment options, and representative results are demonstrated.

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

The severe organ shortage has resulted in increased use of marginal kidney grafts for transplantation. This has triggered interest in alternative storage methods, since marginal grafts especially tolerate cold storage poorly. The technique of normothermic ex vivo kidney perfusion (NEVKP) represents a novel preservation method for kidney grafts prior to &#160;transplantation.

정상 체온<em&gt; 전의 VIVO</em&gt; 신장의 보존을위한 신장 관류는 이식 전에 이식

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a ...

Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality of life when compared to dialysis. Although the last few decades have seen major improvements in patient outcomes following kidney transplantation, the increasing shortage of available organs represents a severe problem worldwide. To expand the donor pool, marginal kidney grafts recovered from extended criteria donors (ECD) or donated after circulatory death (DCD) are now accepted for transplantation. To further improve the postoperative outcome of these marginal grafts, research must focus on new therapeutic approaches such as alternative preservation techniques, immunomodulation, gene transfer, and stem cell administration.
Experimental studies in animal models are the final step before newly developed techniques can be translated into clinical practice. Porcine kidney transplantation is an excellent model of human transplantation and allows investigation of novel approaches. The major advantage of the porcine model is its anatomical and physiological similarity to the human body, which facilitates the rapid translation of new findings to clinical trials. This article offers a surgical step-by-step protocol for an autotransplantation model and highlights key factors to ensure experimental success. Adequate pre- and postoperative housing, attentive anesthesia, and consistent surgical techniques result in favorable postoperative outcomes. Resection of the contralateral native kidney provides the opportunity to assess post-transplant graft function. The placement of venous and urinary catheters and the use of metabolic cages allow further detailed evaluation. For long-term follow-up studies and investigation of alternative graft preservation techniques, autotransplantation models are superior to allotransplantation models, as they avoid the confounding bias posed by rejection and immunosuppressive medication.

Porcine models of organ transplantation provide an important platform to study mechanisms of organ preservation. This article describes a heterotopic porcine renal autotransplantation model, which allows investigating new approaches to improve the outcome of transplantation using marginal kidney grafts.

돼지 모델에서 이소성 신장자가 이식 : 단계별 프로토콜

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality ...

Nerve-sparing Mid-urethral Obstruction (NeMO) in Female Small Rodents

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal model to investigate bladder remodeling in pBOO are female rodents undergoing partial obstruction at the proximal urethra. Variability in the degree of obstruction and animal mortality are major concerns with proximal obstruction. Furthermore, dissecting around the proximal urethra and bladder neck jeopardizes bladder innervation.
We developed a nerve-sparing mid-urethral obstruction (NeMO) model for pBOO avoiding the disadvantages of the traditional model. We approached the urethra just inferior to the pubic symphysis, which obviated the need for laparotomy as well as for dissection in this area; also, the striated urethral sphincter remained untouched. We performed NeMO in female Sprague-Dawley rats (12 obstructions, 6 sham animals) as well as in female C57/bl6 mice (20 obstructions, 18 sham animals). After two weeks, we evaluated bladder function, bladder mass, and body mass.
We had no mortalities among obstructed- or sham-operated female rats; as described for the traditional proximal pBOO-method, we tied the suture around the proximal urethra and a temporarily placed 0.9 mm metal rod. NeMO induced an 85% increase in bladder mass after two weeks, average residual urine volume was 0.4 mL in partially obstructed rats while only 0.03 mL in sham animals. In mice, we tested 3 sizes of cannulas that we placed along the urethra when tying the suture. We found that using a 27-gauge cannula resulted in over 50% animal mortality; placing the 25-gauge cannula did not yield the desired response in increasing bladder mass; utilizing a 26-gauge cannula yielded favorable results with minimal animal mortality (1/8) yet a significant 2-fold increase in bladder mass.

Nerve-sparing Mid-urethral Obstruction NeMO in Female Small Rodents

Traditional modeling of partial bladder outlet obstruction in rodents is fraught with animal mortality. A denervation injury from dissection around the proximal urethra and bladder neck is also of major concern. We developed and evaluated a safe and reliable mid-urethral obstruction model, avoiding the shortcomings of the traditional model.

여성 작은 설치류에 신경을 살려주는 중간 요도 폐색 (네모)

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal ...