eoe sub articles

EoE Sub Articles

1. Construction of the HIT Device 
<ol>
	<li>Attach the spring to the board using two clamps and four screws (Figure 1A). Center the clamps relative to the width of the board and butt them up against one another with the outer clamp flush with the edge of the board. Prior to attaching the clamps, bend them using pliers to fit tightly over the spring. 
	NOTE: See Table 1 for descriptions of the materials required for constructing the HIT device. The clamped end of the spring should be 1/8 inch (3.2 mm) from the edge of the board, and the free end should extend over the board by 3/4 inch (19 mm). Adjust the spring so that it lies parallel with the length of the board.</li>
	<li>Wrap the free end of the spring once around with the adhesive strip of Velcro loops. The outer edge of the Velcro should be flush with the end of the spring. The Velcro is important because it is used to secure the vial to the spring by creating a tight compression fitting. The Velcro also permits easy connection and removal of vials, allowing many vials to be processed in a short period of time.</li>
	<li>Place the ice bucket cover upside down, centered, tight against the wooden board. Orient the raised region of the ice bucket cover such that the long edge is parallel to the width of the board. Note that the raised region of the ice bucket is 1/2 inch (13 mm) higher than the wooden board, so that when a vial is attached to the spring the spring will not lie flat on the board.</li>
	<li>Slide the whole device against a fixed object such as a wall, so that the ice bucket cover is wedged between the board and the object and does not move.</li>
	<li>Tape the paper protractor to the bottom of a cardboard fly vial tray and stand it on edge against the length of the board so that the 90&#176; mark is aligned with the spring when it is pulled back to a perfectly vertical position.</li>
</ol>
2. Operation of the HIT Device
<ol>
	<li>Place between 1 and 60 CO2-anesthetized flies in an empty vial and stopper the vial using a tight-fitting cotton ball.</li>
	<li>Confine the flies to the bottom 1 inch (2.5 cm) of the vial by pushing the cotton ball into the vial until it is 1 inch (2.5 cm) from the bottom. It is helpful to draw a line on the vial at the 1-inch (2.5 cm) mark. Note that confining flies to larger or smaller regions of the vial can affect the severity of phenotypes.</li>
	<li>Wait 5 min for the flies to recover mobility from the CO2. Note that it is not known whether 5 min is sufficient to completely remove the effects of CO2.</li>
	<li>Insert the end of the spring into the vial until the inner edge of the Velcro is flush with the top of the vial (Figure 1B). When the spring is lying flat, 1 inch (2.5 cm) of the vial should overlap the raised region of the ice bucket cover. Vials can be reused many times.</li>
	<li>While sitting, hold the vial at the Velcro region using the thumb and forefinger of your left hand. Hold the board tight to the benchtop using your right hand. Alternatively, use C-clamps to hold the board tight to the benchtop.</li>
	<li>Pull the spring perfectly straight back to the desired angle. Release the spring. Allow the spring to come to a complete stop.</li>
	<li>Remove the vial from the spring and allow the flies to recover for &#8805;5 min. Subject the flies to another strike or transfer the flies to a vial with fly food. 
	NOTE: A variety of assays can be used to evaluate the phenotypic effects of strikes from the HIT device. For example, effects on longevity can be determined by analyzing the percentage of flies that survive at times after injury, effects on brain morphology can be determined by histological analysis of the head, and effects on gene expression can be determined by quantitative analysis of mRNA levels.</li>
	<li>Determine effects of the procedure that are not due to strikes by identically treating control flies that are not subjected to strikes. Wear hearing protection because the impact of the vial against the ice bucket cover produces a loud noise.</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Zinc plated compression spring</td>
			<td>The Hillman Group</td>
			<td>540189</td>
			<td>9 7/8 in (length), 15/16 in (outer diameter), 0.12 in (wire size)</td>
		</tr>
		<tr>
			<td>Wooden board</td>
			<td></td>
			<td></td>
			<td>9 in (length), 6.5 in (width), 0.75 in (height)</td>
		</tr>
		<tr>
			<td>Clamps</td>
			<td>Sigma Electrical Manufacturing Corporation</td>
			<td>49822</td>
			<td>3.10 in (length), 0.68 in (width), 1.11 in (height), EMT Two Hole Straps, click on type for 1 inch steel EMT conduit</td>
		</tr>
		<tr>
			<td>Loop half of self-adhesive velcro</td>
			<td></td>
			<td></td>
			<td>3 in (length), (3/4 in width)</td>
		</tr>
		<tr>
			<td>Polyurethane ice bucket cover</td>
			<td>Fisher Scientific</td>
			<td>02-591-45</td>
			<td>9 1/8 in (length), 9 1/8 in (width), 1 1/4 in (height)</td>
		</tr>
		<tr>
			<td>Plastic fly vials</td>
			<td>Applied Scientific</td>
			<td>AS-510</td>
			<td>3 11/16 in (height), 1 1/16 in (inner diameter), 1 1/8 in (outer diameter)</td>
		</tr>
		<tr>
			<td>Large cotton balls</td>
			<td>Fisher Scientific</td>
			<td>22-456-883</td>
			<td></td>
		</tr>
		<tr>
			<td>Paper protractor</td>
			<td></td>
			<td></td>
			<td>10 in (diameter)</td>
		</tr>
	</tbody>
</table>

a method to inflict closed head traumatic brain injury in drosophila

Source: Katzenberger, R. J., et al.,&#160;<a href="https://app.jove.com/v/52905">A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila.</a> J. Vis. Exp. (2015).
The video demonstrates the use of a High-Impact Trauma (HIT) device to induce closed-head traumatic brain injury (TBI) in Drosophila. TBI causes mechanical brain trauma affecting neuronal function, which over time leads to neurodegeneration and mortality.

<img alt="Figure 1" src="/files/ftp_upload/23123/23123_Figure1.jpg" /> 
Figure 1: Diagram of the HIT device. (A) Illustration of the top view of the HIT device in the resting position. (B) Illustration of the side view of the HIT device. The spring is shown in the resting position (dark red) and deflected to 90&#176; (light red).

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Source: Katzenberger, R. J., et al.,&#160;A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila. J. Vis. Exp. (2015).
The video ...

Take anesthetized adult Drosophila flies in an empty vial.
Insert a cotton ball into the vial to confine the flies at the bottom, ensuring a stronger impact when inflicting injury.
Once the flies recover from anesthesia, attach the vial to the spring of the high-impact trauma device.
Rest the end of the vial on a raised platform. Then, pull back the spring to an angle and release it.
The vial strikes the platform, causing the flies to collide against the vial walls repeatedly.
The drosophila brain strikes against the head capsule, mimicking closed-head traumatic brain injury, or TBI, without causing external damage.
Allow the flies to recover, and transfer them to a vial with food.
The injury causes damage to neuron membranes, impairing neuronal function.
Signaling molecules from the damaged neurons recruit immune cells, leading to progressive degradation of brain tissue, eventually resulting in the flies&#39; death.

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Trauma

36 visualizações. Source: Katzenberger, R. J., et al., A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila. J. Vis. Exp. (2015). The video demonstrates the use of a High-Impact Trauma (HIT) device to induce closed-head traumatic brain injury (TBI) in Drosophila. TBI causes mechanical brain trauma affecting neuronal function, which over time leads to neurodegeneration and mortality. 

Vídeo: A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila - Conceito

Investigating Alterations in Caecum Microbiota After Traumatic Brain Injury in Mice

Increasing evidence shows that the microbiota-gut-brain axis plays an important role in the pathogenesis of brain diseases. Several studies also demonstrate that traumatic brain injuries cause changes to the gut microbiota. However, mechanisms underlying the bidirectional regulation of the brain-gut axis remain unknown. Currently, few models exist for studying the changes in gut microbiota after traumatic brain injury. Therefore, the presented study combines protocols for inducing traumatic brain injury using a lateral fluid percussion device and analysis of caecum samples following injury for investigating alterations in the gut microbiome. Alterations of the gut microbiota composition after traumatic brain injury are determined using 16S-rDNA sequencing. This protocol provides an effective method for studying the relationships between enteric microorganisms and traumatic brain injury.

Presented here is a protocol to induce diffuse traumatic brain injury using a lateral fluid percussion device followed by the collection of the caecum content for gut microbiome analysis.

Investigando alterações na microbiota de caecum após traumatismo cranioencefálico em camundongos

Increasing evidence shows that the microbiota-gut-brain axis plays an important role in the pathogenesis of brain diseases. Several studies also ...

JoVE Journal

Neurociência

Murine Model of Controlled Cortical Impact for the Induction of Traumatic Brain Injury

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the United States. In fact, TBI is a contributing factor to over a third of all injury-related mortality. Nonetheless, the cellular and molecular mechanisms underlying the pathophysiology of TBI are poorly understood. Thus, preclinical models of TBI capable of replicating the injury mechanisms pertinent to TBI in human patients are a critical research need. The controlled cortical impact (CCI) model of TBI utilizes a mechanical device to directly impact the exposed cortex. While no model can full recapitulate the disparate injury patterns and heterogeneous nature of TBI in human patients, CCI is capable of inducing a wide range of clinically applicable TBI. Furthermore, CCI is easily standardized allowing investigators to compare results across experiments as well as across investigative groups. The following protocol is a detailed description of applying a severe CCI with a commercially available impacting device in a murine model of TBI.

Here we describe a protocol for the induction of murine traumatic brain injury via an open-head controlled cortical impact.

Modelo murino de impacto cortical controlado para a indução de traumatismo cranioencefálico

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the ...

Validando a resposta a lesões e avaliações comportamentais em um modelo de rato mTBI

Rat Model of Closed-Head Mild Traumatic Injury and its Validation

Animal models are crucial for advancing our understanding of mild traumatic brain injury (mTBI) and guiding clinical research. To achieve meaningful insights, developing a stable and reproducible animal model is essential. In this study, we report a detailed description of a closed-head mTBI model and a representative validation method using Sprague-Dawley rats to verify the modeling effect. The model involves dropping a 550 g mass weight from a height of 100 cm directly onto the head of a rat on a destructible surface, followed by a 180-degree turn. To assess the injury, rats underwent a series of neurobehavioral assessments 10 min post-injury, including time of loss of consciousness, first seeking-behavior time, escape ability, and beam balance ability test. During the acute and subacute stages following the injury, behavioral tests were conducted to assess motor coordination ability (Beam task), anxiety (Open Field test), and learning and memory abilities (Morris Water Maze test). The closed-head mTBI model produced a consistent injury response with minimal mortality and replicated real-life situations. The validation method effectively verified the model development and ensured the stability and consistency of the model.

Autor em destaque: Avançando na pesquisa de lesões cerebrais traumáticas - um modelo de cabeça fechada para replicação precisa e avaliação rápida 

Here, we present a closed-head mild traumatic brain injury (mTBI) rat model and its validation exhibiting remarkable similarity to human mTBI concerning behavioral manifestations during the acute and subacute stages.

Modelo de Lesão Traumática Leve de Cabeça Fechada em Rato e sua Validação

Animal models are crucial for advancing our understanding of mild traumatic brain injury (mTBI) and guiding clinical research. To achieve meaningful ...

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Transcrição

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Investigando alterações na microbiota de caecum após traumatismo cranioencefálico em camundongos

Modelo murino de impacto cortical controlado para a indução de traumatismo cranioencefálico

Modelo de Lesão Traumática Leve de Cabeça Fechada em Rato e sua Validação

Modelo de Lesão Traumática Leve de Cabeça Fechada em Rato e sua Validação

Modelo de Lesão Traumática Leve de Cabeça Fechada em Rato e sua Validação