JP2010054489A - Method and device for measuring metabolic state of animal individual - Google Patents

Abstract

The present invention relates to the collection of skin gas in small animals, so far no skin gas research on small animals has been found, and this is a completely unknown field. Small animals are also important experimental means for research and development of drug development and disease states. It is easy and easy to measure the progress of small animals.
In order to fix a skin exposed part / local part of an animal individual, the animal individual is inserted into a cylindrical fixture 1, for example, a tail part is exposed to the outside, and a resin sheet is used from the tail part of the animal individual. Collect skin gas. This makes it possible to collect skin gas from experimental animals and understand the relationship between skin gas and pathology of small animals.
[Selection] Figure 1

Description

There are patents regarding the collection of human skin gas (patent numbers: Patent No. 4024817, Patent No. 4024818, Patent No. 4057097), and a collection tool for collection from fingers and arms is disclosed. However, the above patent is limited to human subjects.

The present invention relates to the collection of skin gas in small animals. So far, there is no research on skin gas in small animals, and it is a completely unknown field. Small animals have become an important experimental means for research and development of drug development and disease states. The collection of skin gas for small animals has never been reported and is a completely new method.

The present invention provides a method and means for collecting skin gas from a new small animal corresponding to this unknown field.

As a method of this kind, as shown in Patent Documents 1 to 3, patents relating to humans have been patented, but they were not directly applicable to small animals.
Also, small animals are often used to ascertain the state of illness and to confirm the effects of pharmaceuticals, but this operation takes time and effort.

Patent No. 4024817 Patent No. 4024818 Japanese Patent No. 40570397

The challenge is to keep the small animal individual at rest, collect skin gas from the local skin, and collect skin gas reflecting the metabolic state of the small animal individual.
When anesthesia is performed on small animals, the effect of anesthesia on an individual is large and is not suitable for grasping the metabolic state of an individual. The local skin gas of small animals is preferably collected from protrusions such as the tail, ears, feet, etc., and these can be fixed independently of the activity of the individual. For this reason, developing a fixing method is a problem to be solved.

The skin gas is collected from the local skin surface of the animal individual to grasp the metabolic state of the animal individual.

In order to fix the skin exposed part / local part of the animal individual, it is provided with an insertion plate having an opening for inserting the animal individual into a cylindrical fixture and exposing only the tail part to the outside. A cylindrical fixture is provided so that it can be collected.

Put a rat or mouse into the fixture of claim 2 and use a collection bag (sampling bag) made of a polyfluoroethylene sheet to collect skin gas from its tail, and seal the opening of the collection bag at the base of the tail There is a need for a collection bag that can be made.

A means for collecting skin gas by attaching a sampling bag made of a polyfluoroethylene sheet to the ear of a rabbit is required.

A means for attaching a sampling bag made of a polyfluoroethylene sheet to a rat's foot and collecting skin gas is required.

The challenge is to keep the small animal individual at rest, collect skin gas from the local skin, and collect skin gas reflecting the metabolic state of the small animal individual.

When anesthesia is performed on a small animal, the effect of anesthesia on an individual is large and is not suitable for grasping the metabolic state of the individual.

The local skin gas of small animals is preferably collected from protrusions such as the tail, ears, and feet, and these can be fixed without being influenced by the activity of the individual. For this reason, developing a fixing method is a problem to be solved

The collection bag of the present invention is the development of a collection bag made of a polyfluoroethylene sheet for collecting skin gas from the tail by placing a rat or mouse in a cylindrical fixture, and the opening of the collection bag at the base of the tail. It can be sealed.

Although the present invention will be described with respect to the tail of a rat or mouse, the same applies to the case of measuring a foot instead of the tail, and the same applies to a rabbit. It is what you use. This is a method and apparatus for collecting skin gas by attaching a polyfluoroethylene sheet to the ears and feet of the rabbit.

As described above, the present invention relates to a method and apparatus for collecting skin gas from a local skin surface such as the tail, ear or foot of an animal individual and measuring the metabolic state of the animal individual.

In order to fix the exposed / local part of the animal's skin, it is equipped with an insertion plate that has an opening for inserting the animal individual into a cylindrical fixture and exposing only the tail to the outside. A cylindrical fixture that can be collected and provided with a cylindrical tubular structure or a double-open structure can be fixed non-invasively without giving a strong stress to an individual animal.
Collecting skin gas using a small animal is a well-known fact that a very difficult experimental system can be effectively performed in a short time. In the present invention, the metabolic state of the body can be grasped by collecting skin gas of a small animal. By collecting skin gas non-invasively from the local area of an animal and measuring changes in chemical components contained in the skin gas, it is possible to provide a means for accurately determining the health condition and disease of an individual body. If the method and measuring instrument of the present invention are used, it is possible to grasp the disease and determine the drug effect very economically.

As shown in FIGS. 1 to 4, the rat fixture of Example 1 has a cylindrical shape so that the rat can be easily put into the fixture. The head of the rat is provided with three grooves so that the position can be changed depending on the size of the rat, and a head fixing partition plate (reference numeral 5) is used in the groove. As the tail, a fixing plate (reference numeral 6) having a cut for the tail was used. After the rat was guided into the cylinder, the tail fixing plate was installed, and the tail was moved out of the fixing plate. With this structure, skin gas can be easily collected from the tail. A partition plate is placed in front of the rat so that the rat cannot escape and the length can be adjusted.

The center part of the rat fixture of Example 1 has a notch of 4 and is devised so that blood can be collected from a catheter placed in the ventral jugular vein.

FIG. 4 shows a method of collecting skin gas from the tail by putting the rat in the rat fixture of Example 1. It is possible to extract only the tail from the tail fixing partition and collect skin gas from the tail. Before collecting the skin, the rat's tail is washed clean with distilled water, and then a polyfluoroethylene sheet sampling bag (symbol 10) is attached. The sampling bag has an opening for inserting the tail and a cock for taking gas in and out. To attach the sampling bag to the rat tail, first wrap a resin film with good stretchability around the base of the tail several times, and then wrap the opening of the sampling bag around the base of the tail of the rat. A tail part and a bag made of a polyfluoroethylene sheet are fixed using a resin film and sealed so that skin gas does not leak. For collecting skin gas, the bag made of polyfluoroethylene sheet is washed several times with nitrogen gas, and then 20 ml of nitrogen gas is put into the bag to collect skin gas for 3 minutes. These operations are performed while operating the three-way stopcocks 14-16. The skin gas collected for 3 minutes in the sampling bag was transferred to a sample storage bag (manufactured by polyfluoroethylene sheet) (symbol 13) and then analyzed using gas chromatography.

As shown in FIG. 5, the rat fixture of the second embodiment is configured such that the door can be fully opened so that the rat can be easily put into the fixture. The rat's tail is taken out of the fixture and cut to make it easier to collect skin gas. A partition plate is placed in front of the rat so that the rat cannot escape and the length can be adjusted.

FIGS. 6 and 7 show views when the rat fixture of Example 2 is half-opened and closed. The central part is cut so that blood can be collected from the catheter.

The rat fixture of Example 2 has a mechanism in which a central portion is fixed by a clasp. In a closed state, a catheter from the rat is led out of the rat fixture through the upper incision and blood is collected.

FIG. 8 shows a method of collecting skin gas from the tail by putting the rat in the rat fixture of Example 2. Only the tail part can be taken out from the fixture, and skin gas can be collected from the tail part. Before collecting the skin, the rat's tail is washed thoroughly with distilled water, and then a polyfluoroethylene sheet bag is attached. When wearing the bag, use parafilm to fix it from the tail of the rat and the polyfluoroethylene sheet bag and seal it so that the skin gas does not leak. For collecting skin gas, the bag made of a polyfluoroethylene sheet is washed several times with nitrogen gas, and then 20 ml of nitrogen gas is put into the bag to collect skin gas for 3 minutes. The skin gas collected in the bag was transferred to a sample storage bag and analyzed.

Example 3 examined diabetic rats. A comparison of the concentration of acetone gas in the skin gas collected from the 10-week-old diabetic rat and the rat tail in the subject group is shown in FIG. As can be seen from FIG. 9, the concentration of acetone collected from the tail of diabetic rats was higher than that of the control group.

Example 4 was performed on 10-week-old hypertensive rats. The results of Example 4 are shown in FIG. FIG. 10 shows a comparison of the acetone concentration in the skin gas obtained from the hypertensive rat and the rat tail in the subject group. As can be seen from FIG. 10, the concentration of acetone collected from the tail of hypertensive rats was higher than that of the control group.

Example 5 examined skin gas nitric oxide for 10-week-old diabetic rats. FIG. 11 shows a comparison of rat tail nitric oxide concentrations in diabetic rats and subject groups. As can be seen from FIG. 11, the nitric oxide concentration collected from the tail of diabetic rats was lower than that of the control group.

In Example 6, a comparison between the concentration of acetone gas obtained from the tail and blood components of rats was performed. A correlation is suggested between the glucose component in blood and the concentration of acetone gas.

In Example 7, the rabbit was put into a cylindrical fixture, and gas could be collected from the ear.

As described above, the present invention is designed to enable collection of skin gas in small animals, so far no skin gas research on small animals has been found, and this is a completely unknown field, but small animals useful as experimental animals. Has become an important experimental tool for research and development of drug development and disease states. This process and result measurement can be performed easily and easily, and its utility value is extremely large. The relationship between skin gas and pathology can be clarified using small animals, and this relationship provides a foundation that can be applied to human diseases. This may enable prediction of disease using skin gas. It also has the potential to discover new skin gases and present new health indicators for disease states. The application to the medical industry and the contribution to human health are enormous, and a great social contribution can be made in the future, and a ripple effect on the industry can be expected.

FIG. 3 is a perspective view of a rat required fixture of Example 1. It is a perspective view when a partition plate is installed on the rat fixture of Example 1 and the entire fixture is put in a light shielding case. FIG. 3 is a plan view of a partition plate of the rat fixing device according to the first embodiment. It is a figure of the state at the time of putting the rat in the fixing tool for rats of Example 1, collecting skin gas from a rat tail, and collecting blood simultaneously. It is a perspective view (full open view) of the rat fixture required of Example 2. FIG. It is a perspective view (semi-open view) of the fixing tool for rats of Example 2. It is a perspective view (front closing figure) of the fixing tool for rats of Example 2. FIG. It is the figure of the state which put the rat in the fixing tool for rats of Example 2, and was collecting the skin gas from the rat tail part, and the state which showed the mode that blood was extract | collected with the catheter simultaneously. It is a figure of the comparison of the rat tail acetone density | concentration in the diabetic rat obtained by Example 3, and an object group. It is a figure of the hypertensive rat obtained by Example 4, and the comparison of rat tail acetone density | concentration in an object group. It is a figure of the comparison of the rat tail nitric oxide density | concentration in the diabetic rat obtained by Example 5, and a test group.

Explanation of symbols

1. 1. Cylindrical small animal fixture 2. Head fixing partition installation groove 3. 3. Tail fixing partition plate installation groove 4. Injection needle insertion port for blood collection 5. Head fixing partition plate 6. Tail fixing partition plate Shading case8. Rat (mouse)
9. Rat tail 10. 10. Skin gas sampling bag 11. Gas exhaust syringe in the sampling bag 12. Sampling clean gas 13. Gas bag for storing sample gas Three-way stopcock15. Three-way stopcock 16. Three-way stopcock 17. Blood collection instrument 18. Door 19. Tail fixed plate (length 65mm, width 110mm, thickness 2mm)
20. Fixed head (length 65mm, width 110mm, thickness 2mm)

Claims (4)

A method of measuring the metabolic state of an animal individual by collecting skin gas from the surface of the local skin of the animal individual. In order to fix the skin exposed part / local part of the animal individual, it is provided with an insertion plate having an opening for inserting the animal individual into a cylindrical fixture and exposing only the tail part to the outside. A device that collects and measures metabolic status. 3. An apparatus according to claim 2, comprising a collection resin sheet bag for collecting skin gas from the tail by placing a rat or mouse in a fixture. 3. The apparatus according to claim 2, comprising a resin sheet bag for collecting skin gas, wherein a resin sheet is attached to a rabbit ear.