Method for simulating experimental amyloid cardiopathy in rats
SUBSTANCE: method consists in a single administration of a mixture consisting of a rat's homogenised myocardial tissue - 25%, egg albumin - 25% and Freund's adjuvant - 50%, into old male rats. The method involves administering in an amount of 0.3 ml into 5 injection points: intraperitoneally, into inguinal and axillary regions from the left and right.
EFFECT: being easy to reproduce and cost efficient, the method is effective for creating a model aiming at a possibility of studying the pathogenesis, preventing and treating cardiopathy.
2 dwg, 1 tbl, 1 ex
The invention relates to medicine, experimental biology and can be used to simulate the experimental amyloid cardiomyopathy in animals with the goal of studying the pathogenesis, prevention and treatment of cardiomyopathy.
Cardiopathy, cardiopathia (from the Greek. cardia - heart and pathos - suffering, defeat), the term applied to such changes of heart that are prolonged or persistent pathological condition which is a consequence of previous degenerative or inflammatory processes.
Of particular interest is the problem of amyloidosis of the heart, which is diagnosed in 2.3% of deaths before the age of 50 years, in the age group of 50-70 years it is detected in 30%, in the group of 70-80 years is already at 41%, and in persons who died at the age of 90 years, amyloidosis was found in 71%-90% of cases. In this regard, the aim of our study is to attempt to simulate a more simple way cardiopatico type of amyloidosis to obtain opportunities for further study of the pathogenesis, prevention and treatment of amyloid cardiomyopathy.
Amyloidosis, amyloid degeneration, impaired protein metabolism, accompanied by the formation in the tissue-specific protein-polysaccharide complex - amyloid. The development of amiloidosis associated with the perversion of the protein-synthetic function of the reticulo-�endoteliales system, the accumulation in plasma of abnormal proteins that serve as autoantigens and induce the formation of autoantibodies. As a result of interaction of the antigen with the antibody occurs deposition of particulate proteins involved in the formation of amyloid. Putting off in the tissues, amyloid displaces functionally specialized elements of the body, which leads to the death of this body.
Based on the above homogenized myocardium of rats applied with the aim of launching specific autoimmune reactions of the body, which in combination with amyloidogenic promotes the formation and deposition of amyloid masses in the myocardium.
A method of modeling of experimental amyloidosis in rats, taken by the authors for the prototype (patent of the Russian Federation for the invention 2410761 C1, gabuyeva A. A., Breen V. B., K. M. Kozyrev, registered 09.10.2009, the application 2009137437/14, 09.10.2009 IPC' G09B 23/28), which consists in the introduction of experimental animals equal mixture of native egg albumin and complete adjuvant of frand rate of 0.2 ml in 5 points of injection.
The disadvantages of the prototype is the low efficiency for specific modeling of amyloidosis of the heart.
Also known a method of simulation of experimental amyloidosis in animals, taken for analogue (RF patent for the invention №2306617 C1, Gioia Z. V., Zaalishvili T. V., Kozyrev, K. M., Breen B. V., registered�recorded 18.04.2006, application 2006113109/13, 18.04.2006 IPC' G09B 23/28), which consists in the introduction of the protein preparation subcutaneously. As the protein preparation is administered to Syrian hamsters native bovine plasma through the day from the calculation of 0.025 ml/g of body weight within 60 days of the experiment.
The disadvantages of analog is the need for repeated administration of protein of the drug subcutaneously every other day for 60 days of the experiment, and the relatively small size of the animals, which hinders functional methods of research models. In addition, Syrian hamsters difficult and uneconomical.
There is a way of modeling of experimental amyloidosis in animals, taken by the authors for the analog (RF patent for the invention №2347279 C1, Pukhov, I. W., K. M. Kozyrev, Bryn B. V., registered 27.09.2007, the application 2007135901/14, 27.09.2007 IPC' G09B 23/28), including the introduction of native porcine plasma through the day from the calculation of 0.025 ml/g of body weight within 60 days of the experiment.
The disadvantages of analog is the need for repeated administration of protein of the drug within 60 days through the day, and small size animals, which greatly complicates the study of the functional state of the cardiovascular system.
There is also a method of simulation of experimental amyloidosis in animals, also taken by the authors for analogue (paten� of the Russian Federation for the invention №2446482 C1, Breen V. B., Belikov A. T., K. M. Kozyrev, registered 13.11.2010, the application 2010146365/14, 13.11.2010 IPC' G09B 23/28), which proposes a single administration to Syrian hamsters native human plasma, mixed with the adjuvant of freind in equal proportions. The injection is carried out in a symmetrical axillary and inguinal region, and intraperitoneally at the rate of 0.1 ml in each point.
The disadvantage of this analogue is the low specificity bestselection effects on the myocardium.
The claimed invention is directed to solving the problem, which consists in the development of a new method of modeling of experimental amyloid cardiomyopathy in animals, consisting in the conduct of a mixture consisting of homogenized myocardial tissue of rats (25%), egg albumin (25%) and adjuvant of franda (50%), 0.3 ml in 5 points of injection: intraperitoneally, in the inguinal and axillary region subcutaneously to the left and right.
The solution of this problem provides empowering simulation experimental cardiomyopathy in animals.
The proposed method differs in that for modeling of amyloid cardiopathy injected a mixture consisting of homogenized myocardial tissue of rats (25%), egg albumin (25%) and adjuvant of franda (50%), 0.3 ml in 5 points of injection: intraperitoneally, in the inguinal and axillary region subcutaneously to the left and right.
The manufacturer.�th method is effective, cost-effective and easily reproducible.
According to the information set of essential features that characterize the essence of the claimed invention, is not known, which allows to make a conclusion about conformity of the invention the criterion of "novelty."
Experimental animals (old rats at the age of 18-24 months, weighing 350-400 g) once injected a mixture of homogenized myocardial tissue of rats (25%), egg albumin (25%) and adjuvant of franda (50%), 0.3 ml in 5 points of injection: intraperitoneally, in the inguinal and axillary region subcutaneously to the left and right, which allows extending the modeling capabilities of experimental cardiomyopathy in animals and allows to make a conclusion on compliance with the criterion of "inventive step".
The set of essential features that characterize the invention, in principle, can be repeatedly used in medicine with the result, consisting in an effective and easily reproducible method of simulation of experimental amyloid cardiomyopathy in animals, which allows to make a conclusion about conformity of the invention the criterion of "industrial applicability".
This method is as follows.
To obtain experimental cardiopathy in experimental animals selected Kry�s-males line "Wistar" weighing 350-400 g at the age of 18-24 months, so how exactly senile bradytrophic fabric allows you to raise the issue of modeling cardiopathy. For modeling cardiopathy in experimental animals once injected a mixture of homogenized myocardial tissue of rats (25%), egg albumin (25%) and adjuvant of franda (50%), 0.3 ml in 5 points of injection: intraperitoneally, in the inguinal and axillary region subcutaneously to the left and right.
In control and experimental animals under hypentelium anesthesia investigated hemodynamic and invasive way in terms 60 days of the experiment, after which the animals were killed. The tissue samples are fixed in 10% neutral formalin, followed by the preparation of paraffin sections with a thickness of 5-6 microns. Sections were stained with hematoxylin and eosin, Congo red. The study of slices is carried out in transmitted light with microscope Mikmod-1 under magnification× 80, × 200, × 600.
The essence of the claimed method is confirmed by functionally and morphologically.
In the study of the main indicators of hemodynamics was significantly decreased: cardiac index (SI) and shock index (si) and the increase of specific peripheral vascular resistance (UPSS) compared with the control (tab. 1). Tended to decrease mean arterial pressure (SBP) and heart rate (HR).
|Changes in key indicators of hemodynamics during experimental formation of amyloid cardiopathy|
|Group||Indicator||GARDEN||Cardiac index||Shock index||UPSS||HR|
Example No. 1. Conducted a series of experiments consisting of 2 groups:
1st group (20 rats-males line "Wistar") control
2nd group (20 rats-males line "Wistar") - experienced� - old rats-males (age 18-24 months, body weight 350-380 g) once introduced a mixture consisting of homogenized myocardial tissue of rats (25%), egg albumin (25%) and adjuvant of franda (50%), 0.3 ml in 5 points of injection: intraperitoneally, in the inguinal and axillary region subcutaneously to the left and right.
The morphological study to identify cardiomyopathy subjected to the myocardium and blood vessels.
In the studies provided morphological evidence for the effectiveness of native egg albumin in combination with the complete adjuvant of freind for modeling experimental cardiomyopathy in old male rats.
Histologically in the 2nd group of animals in the myocardium (Fig. 1, 2) when stained with sections of hematoxylin-eosin and Congo red revealed areas of parenchymal dystrophy protein, sometimes passing to lobular collapse and fragmentation of cardiomyocytes with the disappearance of the transverse iscertainly (1). Marked perivascular limfogistiocitarnom infiltration (2), stromal edema (3), the plethora of the vessels of the microvasculature (4). In addition, marked sclerotic changes of stroma. Is the plasmatic impregnation of the walls of the microcirculatory vessels and perivascular areas of plasturgie (5).
When stained with Congo-red marked ex�proposal amorphous masses of amyloid in the walls of blood vessels and stroma of the myocardium (6).
Thus, the results of morphological studies of tissues of experimental animals fed with amyloidogenic equal mixture consisting of homogenized myocardial tissue of rats was 25%, egg albumin 25% and adjuvant of frand - 50%, 0.3 ml in 5 points of injection: intraperitoneally, in the inguinal and axillary region subcutaneously on the left and right show the development of experimental amyloidosis with primary lesion of the myocardium.
Summing up, it should be noted that a single administration of a mixture consisting of a homogenized myocardial tissue of rats (25%), egg albumin (25%) and adjuvant of franda (50%), 0.3 ml in 5 points of injection: intraperitoneally, in the inguinal and axillary region subcutaneously to the left and to the right is an effective way of modeling the experimental cardiopathy.
The method of experimental simulation of amyloid cardiopathy in rats, consisting in a single administration to old rats-males of the mixture consisting of pre-blended fabric of the myocardium of rats was 25%, egg albumin 25% and an adjuvant of frand - 50%, 0.3 ml in 5 points of injection: intraperitoneally, in the inguinal and axillary region subcutaneously to the left and right.
SUBSTANCE: phantom test consists of two parts being a single whole. One part has a constant longitudinal height, while the other part has a continuously varying longitudinal height forming a wedge. On the side of the wedge, there are calibrated notches.
EFFECT: more accurate determination of the bone mineral density by X-ray images formed with the help of a general-application X-ray machine, and simplifying the design.
SUBSTANCE: invention refers to medicine, particularly to gastroenterology, pathologic physiology, and concerns simulating acute pancreatitis. To this effect, the method involves ligaturing the main truck of the excretory duct of the pancreas, administering an aggressive solution into the pancreatic duct system, and removing the ligature. The aggressive solution represents 1% chenodeoxycholic acid with 5% sodium hydrogen carbonate in an equal proportion in an amount of 0.3-0.5 ml at 10-15 mg/kg of the animal's body weight.
EFFECT: method enables developing a model of acute fatty, hemorrhagic or mixed pancreatitis in the animal that makes it applicable for improving the known methods for conservative and radical methods of treating.
SUBSTANCE: invention refers to medicine, particularly to ophthalmology, and concerns simulation in vivo of cortical cataract. That is ensured by a double-side surgical sympathectomy by excising a superior cervical sympathetic ganglion.
EFFECT: by simplicity and effectiveness of modulation, the method provides forming the cortical lens-form opacity, which is morphologically and immunohistochemically identical to lens cell changes in age-related lens opacity in a human.
6 dwg, 2 tbl
SUBSTANCE: during the trial rabbits are given the inhibitor preparation lynestrenol in vivo endogastrically with the daily dose of 110 mkg/kg of animal body weight in the course of 14, 21 or 28 days.
EFFECT: invention makes it possible to use lynestrenol as positive control of lower activity of glycoprotein-P for prediction of inclusion of studied medicinal agents into substrates of a transporter protein.
SUBSTANCE: cecum lymphoma is modelled by the introduction of 2,4,6-trinitrobenzosulfonic acid to a rat in a dose of 0.1-0.15 ml, diluted in 0.1-0.15 ml of a 50% ethanol solution. Introduction is realised into a submucosal layer of the cecum cupola 2-3-times daily for 2-3 weeks.
EFFECT: creation of an adequate model of the cecum lymphoma.
SUBSTANCE: to correct pathologic changes in the condition of viable offspring under a cytostatic impact the medication glutoxim is introduced to female rats in a dose of 50 mcg/kg 5 days before and 5 days after the introduction of the cytostatic medication vepesid. The latter is introduced once intravenously in a maximal tolerable dose, equal to 30 mg/kg. It has been established that glutoxim can be applied as means for the correction of pathologic changes in the viable offspring of rats, obtained from coupling 3 months after the cytostatic impact.
EFFECT: application of glutoxim as the means of corrective therapy makes it possible to increase efficiency and reduce its side effects.
SUBSTANCE: method is realised by the intravenous allogeneic transplantation of multipotent mesenchymal stromal cells (MMSC) and hematopoietic stem cells (HSC) to laboratory mice one hour after irradiation. HSC are obtained from the placenta of female mice at the 14-day gestation term. MMSC are introduced in a dose of 6.5 mln cells/kg, with HSC being introduced in a dose of 400 thousand cells/kg.
EFFECT: invention makes it possible to extend the arsenal of means, capable of providing the regeneration potential of the spleen tissues, as well as to increase the regeneration of main morphometric spleen indices after exposure to a radiation load.
SUBSTANCE: melanin having water-solubility of at least 80% and an paramagnetic centre concentration of at least 8·1017 spin/g is administered orally into the animals having been exposed to the radiation in a dose high enough to cause a spinal radiation injury; melanin is administered after dissolved in distilled water in the effective concentration. Melanin water is used as drinking water for the mice having been exposed to single and fractionated acute radiation, which is able to cause acute radiation disease. Melanin water is taken from the 1st to 30th day following the single radiation, or from the 1st day of the fractionated radiation to the 30th day on completion of the radiation.
EFFECT: higher survival rate, faster recovered haemopoiesis, body weight and orientation and motion activity.
7 tbl, 5 ex
SUBSTANCE: invention refers to medicine, namely to animal's behavioural physiology. The orientational-exploratory and locomotor behaviour in rats is rated with an underlying developing acquisition of a food-producing skill by differentiating an animal's motion path in a batwing-shaped labyrinth. The rating procedure is based on the following forms of motor actions: movement to the labyrinth loci without a food reward; purposive movement to the locus with the food reward; exploratory travelling through all the labyrinth landings; movements to the locus contra lateral with the food reward; multiple movements between the two labyrinth loci; freezing behaviour in the start position.
EFFECT: method enables increasing the investigation reliability that is ensured by a multiple assessment of the central nervous system functions.
2 cl, 1 tbl, 1 ex, 6 dwg
SUBSTANCE: method includes sampling bone blocks with a porous wire material to be analysed, fixation of the bone tissue with the porous wire material, decalcination, degreasing, dehydration, pouring the said tissue, preparation of cuts, staining and histomorphometry. Before or after the fixation of the bone tissue with the analysed porous wire material and before or after decalcination search and fixation of an edge wire element of the porous wire material in the bone block is carried out. Untwisting of the porous wire material from the bone block is performed by the method of pulling out the edge wire element. An analysis of macropreparations is carried out when pore spaces of the porous wire material are emptied. After that, the tissues are sampled from the emptying pore spaces, including the sampling and analysis of intra-pore fragments.
EFFECT: possibility of the qualitative and quantitative estimation of histomorphometric parameters of intra-pore tissues in the entire volume of an implant.
3 ex, 1 dwg
FIELD: medicine, experimental abdominal surgery.
SUBSTANCE: as experimental animals one should apply mongrel dogs of 12-17 kg body weight. Under general anesthesia one should conduct superior-median laparotomy, introduce 3.0 ml 70%-ethanol solution under pancreatic capsule and then laparotomic wound should be sutured up. Manipulation should be performed once. The method provides modeling adequate acute pancreatic inflammation at no side effects being very simple in implementation.
EFFECT: higher efficiency.
FIELD: medicine; medical engineering.
SUBSTANCE: method involves studying transverse longitudinal and rotation stiffness characteristics. The studies are carried out step-by-step from the first order units to complete external fixation apparatus structure. The device has frame and is provided with calibration loads, wire rope, displacement indicators, strip for fastening to loading end of bone imitator fragment, beam for fixing displacement indicators, beams having unit for modeling longitudinal and transverse loadings. The frame is manufactured as parallelepiped. The fixing panel has openings for bone imitator, for fixing external fixation apparatus and yoke connection union and is fixed in end face part of the frame. Beam for fixing displacement indicators has longitudinal slit for fixing the indicators and arranging them on lateral slots in frame base. The beams having unit for modeling rotational, longitudinal and transverse loadings are arranged on lateral frame sides on lateral slots in base.
EFFECT: high vision acuity without applying spectacle-based correction; accelerated treatment course.
2 cl, 16 dwg, 1 tbl
FIELD: experimental medicine.
SUBSTANCE: the present innovation deals with modeling urinary calculosis in rats due to injecting intraperitoneally 60%-glucose solution at 1 ml/100 g animal body weight twice daily for 2 mo. The method is very simple and enables to achieve lithogenesis in 25% experimental animals.
EFFECT: higher efficiency of experimental modeling.
2 dwg, 2 ex, 4 tbl
SUBSTANCE: method involves using Hann diode crystal with proper frequencies of pathogenic microorganisms and cells during their death period or during the stimulating factors action period being applied.
EFFECT: enhanced effectiveness of treatment; wider range of biophysical action types.
3 cl, 1 tbl
FIELD: medicine, experimental physiology.
SUBSTANCE: hypoxia with hypercapnia should be modeled due to creating a closed system of inhaled air circulation. Air enters lungs out of hermetically sealed reservoir and at expiration returns back. The process of recirculation is supplied with an apparatus of artificial pulmonary ventilation. The innovation suggested provides steadiness in development of hypoxia with hypercapnia excluding the development of stressor reaction. Conditions should be created to carry out any manipulations with an animal in the course of an experiment.
EFFECT: higher efficiency of modeling.
5 dwg, 1 ex
FIELD: medicine, stomatology.
SUBSTANCE: the present innovation should be carried out for the purpose to study ethiology and pathogenesis of parodontitis. One should affect with emotional stress in experimental animals (mature rats) due to placing 10-11 experimental animals into the cage at area of 0.018 sq. cm/animal. Before placing into the cage one should create artificial dental plaque around the cervix of the upper and lower incisors with the help of stomatological cement for every experimental animal. In the course of modeling all experimental animals should eat paste-like food. The method enables to shorten terms for obtaining the model desired and increase its similarity with pathomorphological manifestations of human parodontitis.
EFFECT: higher efficiency of investigation.
FIELD: medical equipment.
SUBSTANCE: device has input first variable resistor, capacitor and permanent resistor. Permanent resistor is connected to arm second and third variable resistors. Second ends of variable resistors are connected with motionless contacts of polarized relay. Movable contact of relay is connected to common bus. Input of device is connected to amplifier which has output connected with control wiring of polarized relay. Second end of wiring is connected with common bus. Device is intended for electrical modeling of balanced and misbalanced conditions of acupuncture point at electropunctural action with unlike-poled signals due to liquidation mutual errors at any circuit of opposite arms of the device. Values of active resistances can be installed independently at any arm of device.
EFFECT: increased precision.
FIELD: experimental medicine.
SUBSTANCE: laboratory animals should be once injected intraperitoneally or intravenously with phenylhydrazine at the dosage of 100-150 mg/kg.
EFFECT: higher efficiency.
1 cl, 4 ex, 2 tbl
FIELD: medicine, experimental biology, ecology, toxicology.
SUBSTANCE: at studying the mechanisms of heavy metals toxic action, in particular, cadmium upon renal function, it is suggested to introduce cadmium sulfate solution into stomach once daily for 2 mo at the dosage of 0.5 mg/kg, on conversion to metal, where cadmium corresponds to 0.5 mg per 1 ml solution. The present innovation enables to study the pathology in dynamics of development and elaborate and searching preparations for treating and preventing chronic toxic nephropathy.
EFFECT: higher efficiency.
1 ex, 3 tbl
SUBSTANCE: method involves exposing cell or cell group to external power source. At least two electrodes are introduced before treating the cells. One of electrodes is set on cytoplasmatic external cell membrane surface and the other one cell membrane and membrane potential value is measured. External electric voltage source is connected to the introduced electrodes oppositely in polarity with cell membrane potential difference value being not less than cell membrane potential.
EFFECT: enhanced effectiveness in building cell damage model by means of energy burst and death.