Method for predicting hallucino-delirious forms of psychoses of cerebro-atherosclerotic genesis

FIELD: medicine, psychiatry.

SUBSTANCE: one should isolate DNA out of lymphocytes of peripheral venous blood, then due to the method of polymerase chain reaction of DNA synthesis one should amplify the fragments of hSERT locus of serotonin carrier gene and at detecting genotype 12/10 one should predict the risk for the development of hallucino-delirious forms of psychoses of cerebro-atherosclerotic genesis.

EFFECT: more objective prediction of disease development.

3 ex

The present invention relates to medicine, namely to psychiatry, and can be used to predict the development of hallucinatory-delusional forms of psychosis cerebral atherosclerotic Genesis.

The study of psychotic disorders in patients with cerebral atherosclerosis represents an actual problem of modern health care. This is because in recent years, in parallel with the increase in the share of older age groups in the human population, an increasing number of elderly patients with psychotic disorders cerebral atherosclerotic Genesis. The significance of this problem is determined also by the diversity of clinical manifestations of psychotic disorders in patients with cerebral atherosclerosis, providentially their current high level of hospitalization and disability of patients and, accordingly, a significant economic burden to society. Therefore, the development of modern methods for predicting psychotic disorders cerebral atherosclerotic Genesis, in particular hallucinatory psychosis-crazy case, as in patients with cerebral atherosclerosis and healthy people is extremely important. Application of the developed method will enable identification of people with a high likelihood of galls Naturno-delusional forms of psychosis cerebral atherosclerotic Genesis for the formation of groups at high risk of occurrence of this pathology and timely, targeted interventions for the prevention of predicted violations.

A known method for predicting the development of hallucinatory-delusional forms of psychosis cerebral atherosclerotic Genesis on the basis of determining the severity of cardiac and cerebral atherosclerosis and abnormalities in the hemostatic system [Gorodilova H.E. Mental disorders in patients with cerebral atherosclerosis and some indicators of the system of coagulation: author. dis.... Kida. the honey. Sciences. - Kazan, 1975. - 20 C.]. However, this method of predicting cerebral atherosclerotic hallucinatory-delusional forms of psychosis is used in patients with already developed atherosclerotic changes and may not be used among healthy people.

The prototype of the present invention is a method for predicting the development of hallucinatory-delusional forms of psychosis cerebral atherosclerotic Genesis on the basis of the evaluation of the whole complex of risk factors (corresponding to gender, age, family history of vascular disease atherosclerotic Genesis and various mental illnesses, current and long-standing exogenous hazards, such as alcoholism, craniocerebral trauma, infection, psychogenic, somatogen, complications of the atherosclerotic process and others)found among obledo the most of persons through a survey and analysis of medical documentation [Grigoriev BC The ratio of endogenous and exogenous factors in the origin and course of psychotic disorders in patients with cerebral atherosclerosis. // Ukr. neuropathol. and the psychiatrist. - 1992. No. 3. - Pp.96-98. Grigor'ev V.S. Prevalence, clinical variants and for neuropsychiatric disorders in patients with cerebral atherosclerosis (clinical-epidemiological analysis): author. dis.... Kida. the honey. Sciences. - SPb., 1992. - 31 S.]. The disadvantage of the presented method is its bias and inaccuracy, because the process of collecting and interpreting historical information in order to identify the listed risk factors is subjective and is determined by the personal characteristics of the subject and the level of professionalism of the doctor.

The objective of the invention was to develop an objective, informative predictor of the development of hallucinatory-delusional forms of psychosis cerebral atherosclerotic Genesis, allowing to estimate the risk of them as in patients with cerebral atherosclerosis and healthy people.

The technical result when using the invention - improving forecast accuracy. This technical result is achieved that selected by the method of phenol-chloroform extraction of genomic DNA from lymphocytes of 10 ml of peripheral venous blood of conduct Ana is from polymorphic locus hSERT gene vector serotonin by polymerase chain reaction (PCR) DNA synthesis. After amplification, the DNA fragments separated by electrophoresis in 6% nondenatured polyacrylamide gel and analyzed under UV light on a UV-transilluminator. When detecting genotype 12/10 locus hSERT gene carrier of serotonin in the surveyed person's risk of hallucinatory-delusional forms of psychosis cerebral atherosclerotic Genesis increases 2.98 times.

The method is as follows. Amplification of polymorphic gene vector serotonin (hSERT) is carried out in the reaction mixture, containing about 0.1-1.0 microgram of genomic DNA with 0.1 units of Taq polymerase, 0.25 μm of the appropriate oligonucleotide primers: 5’-GTCAGTATCAACAGGCTGCGAG-3’ and 5’-TGTTCGTAGTCTTACGCCAGTG-3' and 250 μm of each deoxynucleotide in 25 ál single PCR buffer containing 67 mM Tris-HCl, pH 8.8; 6,7 mM MgCl₂; 16,6 mM (NH₄)₂SO₄; 0,01% Tween-20. Mode the following amplification: initial denaturation at 95°C for 3 min, 30 cycles including annealing of primers at 58°C - 30 sec, DNA synthesis at 72°C - 1 min, denaturation at 95°C - 30 sec. Amplificatoare DNA fragments separated by electrophoresis in 6% polyacrylamide gel, stained with the bromide Aditya and identify under ultraviolet light. Electrophoresis of DNA fragments is carried out in 1*Tris-borate buffer (1*TBE) (0,089 M Tris-HCl pH 7.8; 0,089 M boric acid is the same; of 0.002 M EDTA with pH 8.0) in a vertical plates at a constant voltage of 300 volts for 1.5 hours after a 20-minute Preparata. Before application to the gel samples are mixed in a ratio of 1:5 with paint containing 0.25% bromophenol blue, 0.25% of xylazine, 15% pecola. As a marker of molecular weight using DNA phage λ, restricion > PST, or plasmid pBR322, restricion Nae. After electrophoresis the gel is stained with a solution of bromide Aditya (0.1 ág/ml in 1*TBE) for 10 minutes, washed with water and examine in ultraviolet light on a UV-transilluminator at a wavelength of 312 nm.

Set significantly higher frequency of genotype 12/10 locus hSERT gene vector serotonin in patients with cerebral atherosclerosis with hallucinatory-delusional forms of psychosis (N=41), compared with patients with cerebral atherosclerosis with nonpsychotic forms of mental disorders (N=50)and control group (N=96): 71%; 26% and 45% (respectively, X²=16,382, R_F=0,0001 and X²=6,748, R_F=0,0084). Calculating genetic risk of hallucinatory-delusional psychosis cerebral atherosclerotic Genesis in the presence of genotype 12/10 locus hSERT gene vector serotonin is performed on the basis of the calculation of the odds ratio is the Odds ratio (OR), reflecting on the relative risk of developing specified psychotic pathology formula OR=(a+0,5)(d+0,5)/(b+0,5)(c+0.5), and where a is the frequency of the genotype in a sample of patients; b - frequency of the genotype in the control sample; C=1 as the sum of the frequencies of other genotypes in a sample of patients; d=1-b is the sum of the frequencies of other genotypes in the control sample; the parameter of 0.5 in the formula is used when one of the numbers a, b, C, d is zero or is less than three. High risk of developing psychotic disorders ascertain when OR>1. In our study, OR=2,98 (CI=1,36-6,52) in the presence of genotype 12/10 locus hSERT gene vector serotonin. Genotype 12/10 locus hSERT gene vector serotonin is a diagnostic genetic marker hallucinatory-delusional forms of psychosis cerebral atherosclerotic Genesis and can be used to identify groups of individuals with high risk of cerebral-atherosclerotic hallucinatory psychosis-crazy case as among patients with cerebral atherosclerosis in the early stages of this disease, and among healthy individuals in order to conduct effective preventive measures.

Example 1.

Patient L., 67 years old, retired for age, was under surveillance with 20.04.2000 year about the cerebral arteriosclerotic psychosis (hallucinatory-delusional syndrome).

Upon receipt complained of headaches, dizziness, noise in the head, loss of memory bessonnica.

History: hereditary cardiovascular disease is not burdened brother the patient is suffering from schizophrenia. Born in a working class family, he was developing normally. He graduated from high school, then College library, in the future, successfully worked as a librarian, then - head of the library. She was married. Her husband died nine years ago, more married not issued. Has a son, a daughter and three grandchildren. Lives with children separately, but the children visit the mother and care for her. The nature of anxious-insecure, impressionable, prone to unreasonable fears. Of illness notes colds, malaria, pneumonia, chronic bronchitis, chronic gastroduodenitis, chronic pancreatitis, cholelithiasis. In 1991, at the age of 58 years for the first time has addressed to the neuropathologist with complaints of a headache, noise in the head and the ears, irritability, fatigue, tearfulness, forgetfulness, insomnia. She was diagnosed sec;Cerebral atherosclerosis (zerebrasteniceski syndrome)sec;. Were outpatients. In subsequent years there was a sec;sec; Serebryanicheskaya symptoms. In the last two years, the patient had begun to deteriorate memory on current events, increased irritability, became short-tempered, extremely distrustful, suspicious, Trevor is Oh. At the end of December 1999 after a conflict with neighbors on the landing was clearly hear their sec;sec; panadero and threatening. Cursed with the sec;sec; through the wall. A few months later came the feeling of a strong smell of gas in the apartment. Was sure that the gas is allowed neighbors through the vents and cracks. Shut down all ventilation openings with polyethylene, were coated plinth. Repeatedly appealed to the police with complaints of neighbors required assistance. During the week before admission to hospital increased anxiety, became very tense, you have a feeling of surveillance by neighbors, expressed unfounded fears. Stated that he has repeatedly seen the neighbors on our balcony, there was a feeling that the neighbors affect her laser beams, want to get rid of it with the purpose of taking possession of her apartment. Noted the loss of things in his apartment. Stopped to leave the house. Argued that the neighbors keep her grandson in the basement, bullied. Heard sec;sec; neighbors, crying grandson and his cries for help from the basement. Night 19.04.99, armed with an axe, went to the entrance of the house and tried to break open the cellar door to free their grandson was excited, screaming, calling for help. Specialized psychiatric brigade was delivered to the Republic of ublicans psychiatric hospital affiliated with the Russian.

Somatic status: correct build, low power. The skin is clean, dry, flabby with a yellowish tinge. Identify lipid arc corneas. Temporal vessels sclerotic (fibrous) and convoluted. Muffled heart sounds, rhythmic, systolic murmur on the apex of the heart. Percutere border of the heart is extended to the left. Blood pressure of 145/90 mm RT. senior rhythmic Pulse 66 beats per minute. In light of harsh breathing. Tongue coated thick white coating. The abdomen is soft, there is some pain at the projection point of the gall bladder. The symptom of “tapping” is negative. No edema. Physiological functions have not been breached.

Neurological status: palpebral fissure D>S, light cramps, lethargy pupillary reaction to light, weak convergence, adaptive nystagmus, slight facial asymmetry, deviation of the tongue to the left. Strengthening the tendon and periosteal reflexes are reflexes of oral automatism. Instability in the Romberg, finger tremor of the outstretched hands, pomahivaya when performing planosol and palacecasino samples.

Mental status: extremely anxious, restless, jumps up from his chair and starts to walk around the office. Focus location, self, and in others faces right, time is not accurate. Contact formally available. Attention accentue owano on painful experiences. Voluble, talking loud. To questions posed by the doctor responsible for the creature. Answers accompanies active gestures. Outraged by the behavior of the neighbors, who allegedly at her every act with the purpose of taking possession of her apartment sec;operate laser sec;trying to poison, sec;let the gas, chlorophos in cracks on the floor and ventilation openings with sec;. Argues that the sec;in the basement of her home is now sec;, says sec;heard his cries and shouts about sec;. With unwavering confidence reports that the sec;it was sec;. Statements are not amenable to correction. Thinking a few slow, thorough. Constantly hears sec;sec;belonging to the neighbors, listens to them, takes them into the conversation. sec;sec; then loud, then move on to the whispers are extraprice and are threatening. Willing to transfer their contents sec;neighbors threaten to kill me and my sec;. The memory is reduced to events

the last years. Impaired memory. Past events remembers quite well. Affective tense, mood reduced. When the manifestation of attention weak. Intelligence is somewhat reduced. Critical attitude to his painful condition no.

Conclusion ophthalmologist: OD - Mature cataract, OS - pronounced sclerosis setca the key, the blood flow is uneven.

Electrocardiography: the electrical axis of the heart rejected the left. Sinus rhythm, heart rate of 68 per minute. Violation vnutrizheludochkovoj conduction, repolarization processes in the lateral parts of the type of the smoothed spike So the Predominance of bioelectric potentials of the myocardium of the left ventricle.

Chest x-ray: right lung has dense foci. The fibrous roots of the lungs condensed, expanded. The diaphragm is movable. Heart expanded at the expense of left ventricular hypertrophy. The aorta sealed, sclerotic (fibrous).

Electroencephalography: prominent diffuse changes of bioelectric activity of the brain, predominantly in the temporal parts of the hemispheres involving nonspecific median formations, θwave on the background of common βrhythm, isolated groups αwaves and bilateral synchronous epileptic polymorphic activity.

Rheoencephalography: marked changes of rogramme on atherosclerotic type.

Laboratory studies: changes of indicators of the General analysis of blood fell within the identified comorbidity. Urine analysis within normal limits. Total cholesterol - 7.22 mol/l, triglycerides - to 4.52 mol/l, β-lipoprotein - 5,42 mol/l Total protein - 73 g/l, total bilirubin of 20.4 µmol/l, sugar - 4,2 mmol/L. PETIT 120%.

The results of the s genotyping. To identify one of the possible genotypes locus hSERT gene vector serotonin, the patient was taken 8 ml of venous blood with subsequent DNA isolation and amplification

polymorphic gene vector serotonin (hSERT) in the reaction mixture, containing about 0.1-1 µg of genomic DNA with 0.1 units of Taq polymerase, 0.25 μm of the appropriate oligonucleotide primers: 5-GTCAGTATCAACAGGCTGCGAG-3’ and 5’-TGTTCGTAGTCTTACGCCAGTG-3’ and 250 μm of each deoxynucleotide in 25 ál single PCR buffer. Electrophoresis of DNA fragments were performed at a constant voltage of 300 volts for 1.5 hours after a 20-minute dielectrophoresis. After electrophoresis the gel was stained with a solution of bromide Aditya for 10 minutes and analyzed under ultraviolet light on a UV-transilluminator. In the examined patient study of locus of hSERT gene vector serotonin revealed the presence of genotype 12/10.

Conclusion: we observed the patient's genetic testing on a subclinical level of cerebral atherosclerosis would determine high risk of hallucinatory-delusional forms of psychosis and time to take preventive measures to prevent the development of the cerebral atherosclerosis and related psychotic disorders delusional-crazy case.

When is EP 2.

Patient K., 62 years old, retired, was under surveillance with 12.04.2000 year about the cerebral atherosclerosis (zerebrasteniceski syndrome).

Complained of fatigue, irritability, tearfulness, headaches, dizziness, noise in the head, loss of memory and insomnia.

History: the heredity of mental illness is not burdened. The mother suffered from atherosclerosis of the brain in old age suffered a cerebral ischemic stroke, died from liver cancer. Father died of lung abscess. The elder brother is sick ischemic heart disease. Younger sister suffers from hypertension.

Pregnancy and childbirth from the mother, as well as the early development of the patient without features. He graduated from high school. Drafted into the army. After serving in the army, graduated from the school of accountancy, then worked as an accountant at the factory. Was married. Since 1997 widower. Has an adult son and daughter. Lives with her daughter, son-in-law and grandchildren. Relations in the family friendly. By nature shy, timid, insecure, indecisive, anxious. Of illness notes frequent sore throat, catarrhal bilateral otitis, dysentery, osteochondrosis of the lumbosacral spine, chronic cholecystitis, chronic pyelonephritis. Of surgical interventions calls tonsillectomy, AP is endacraig. Do not smoke, do not drink alcohol. Considers himself ill in 1997 after the death of his wife. Appeared tearfulness, irritability, fatigue, began to be troubled with headaches, memory went down, fell sharply efficiency, disrupted sleep. Was seen by the neurologist about sec;initial period of cerebral atherosclerosis (zerebrasteniceski syndrome)sec;. Treatment received on an outpatient basis. After courses of therapy subjectively noted some improvement. From 1998 to 2000, the state hesitated. In March 2000 after a stressful situation (the son was in a car accident) condition has deteriorated: irritability became extremely pronounced, tearfulness appeared for no reason, increased self-confidence, headaches have acquired a permanent character, appeared dizziness, has become absent, significantly decreased memory, sec;started all sec;, disturbed sleep (asleep easily, sleep anxiety, superficial, short). The rest did not bring continued improvements, decreased activity, lost interest in life, has been lost.

Somatic status: correct build, low power. The skin is pure white. Blood vessels a little condensed. Muffled heart sounds, rhythmic, systolic murmur on the apex of the heart. Percutere gr is nice heart enlarged to the left. Blood pressure is 135/85 mm RT. Art. the rhythmic Pulse of 70 beats per minute, satisfactory filling and voltage. In the lungs vesicular breathing, abnormal noises are not heard. The root of the tongue coated with white bloom. The abdomen is soft, palpation, a slight tenderness in the right hypochondrium. Symptom sec;sec; negative. No edema. Urination and stool is not broken.

Neurological status: palpebral fissure equal, reaction of pupils to light sluggish, easy smoothness of the right nasolabial fold. The increased tendon reflexes. Notes rocking in the Romberg position, trembling of fingers of the outstretched hands. Expressed vasomotor lability.

Mental status: oriented to place, time, self, others and the situation correctly. Willingly comes into contact. During the conversation, worried. Has many health complaints. If statement looks at the paper, where they recorded. Giving information about yourself, easily crying. Notes that can't watch television and read, because he tires easily, thus increasing headache, there is a giddiness. During the conversation quickly gets tired, irritable, and weak. Looking for help, sympathy, hope for treatment. Attention exhaustible. There is a decrease in memory on the current account of the Oia. Memory for past events stored. Associative processes are slowed down. Intelligence without signs of slowing. The mood is unstable. Critical to your painful condition.

Conclusion ophthalmologist: nipples optic nerves pink, with clear boundaries, moderately expressed sclerosis of retinal vessels.

ECG: sinus rhythm, heart rate of 67 per minute. Not pronounced diffuse changes of the myocardium with a primary lesion of the left ventricle.

Chest x-ray: pathology of the lung is not revealed. The diaphragm is movable, highly situated. Heart increased.

the expense of the hypertrophied left ventricle. The aorta is diffusely enlarged, expanded.

Electroencephalography: a stable changes of bioelectric potentials of the brain diffuse nature with its reduction in the right hemisphere. Signs of mild dysfunction of the median formations brain.

Rheoencephalography: change REG-curve atherosclerotic type moderately expressed.

Laboratory studies: changes of indicators of the General analysis of blood and urine were within the scope of the identified comorbidity. Total cholesterol - to 7.25 mol/l, triglycerides - 4,11 mol/l, β-lipoproteins is 5.54 mol/l Total protein 68 g/l, total bilirubin - 10.9 mmol/l, sugar - 3,8 IMO the b/L. PETIT 111%.

The results of genotyping. To define one of the possible genotypes locus hSERT gene vector serotonin, the patient was taken 8 ml of venous blood with subsequent DNA isolation and amplification of polymorphic gene vector serotonin (hSERT) in the reaction mixture, containing about 0.1-1 µg of genomic DNA with 0.1 units of Taq polymerase, 0.25 μm of the appropriate oligonucleotide primers: 5-GTCAGTATCAACAGGCTGCGAG-3’ and 5’-TGTTCGTAGTCTTACGCCAGTG-3’ and 250 μm of each deoxynucleotide in 25 ál single PCR buffer. Electrophoresis of DNA fragments were performed at a constant voltage of 300 volts for 1.5 hours after a 20-minute dielectrophoresis. After electrophoresis the gel was stained with a solution of bromide Aditya for 10 minutes and analyzed under ultraviolet light on a UV-transilluminator. A study of locus of hSERT gene vector serotonin in the examined patient revealed the presence of genotype 12/10.

Conclusion: since the presence of genotype 12/10 locus hSERT gene vector serotonin increases the risk of hallucinatory-delusional psychosis with cerebral arteriosclerosis in 2,98 times, to prevent the development of this psychotic pathology of the examined patient is necessary to increase the multiplicity of anti-relapse treatment of cerebral atherosclerotic disorders is also possible to limit the impact of the above pathogenic significance of external influences, contributing to the pathological expression of the gene.

Example 3.

Female 34 years - a blood relative of the patient, who was undergoing treatment at the national hospital MOH regarding cerebral arteriosclerotic psychosis (hallucinatory-delusional syndrome).

Health complaints were not filed. The burdened heredity as cardiovascular disease and mental illness. The mother suffers from ischemic heart disease and hypertension, the father of atherosclerosis of the coronary and cerebral vessels, complicated hallucinatory-delusional psychosis. Born in the family of an employee, developed normally. He graduated from high school, then medical school, works as a medical assistant. Married. Has a daughter. By nature reserved, sensitive, easily vulnerable, touchy. Of illness celebrates children's infections, colds, inosinic, felon. When clinical and paraclinical examination pathology of the internal organs was not found.

The results of genotyping. To define one of the possible genotypes locus hSERT gene vector serotonin in the examined was taken 8 ml of venous blood with subsequent DNA isolation and amplification of polymorphic gene vector serotonin (hSERT) in the reaction mixture, which contains Asa about 0.1-1 µg genomic DNA, of 0.1 units of Taq polymerase, 0.25 μm of the appropriate oligonucleotide primers: 5-GTCAGTATCAACAGGCTGCGAG-3’ and 5’-TGTTCGTAGTCTTACGCCAGTG-3’ and 250 μm of each deoxynucleotide in 25 ál single PCR buffer. Electrophoresis of DNA fragments were performed at a constant voltage of 300 volts for 1.5 hours after a 20-minute dielectrophoresis. After electrophoresis the gel was stained with a solution of bromide Aditya for 10 minutes and analyzed under ultraviolet light on a UV-transilluminator. A study of locus of hSERT gene vector serotonin in the examined women revealed genotype 12/10.

Conclusion: due to the fact that the carrier genotype 12/10 locus hSERT gene vector serotonin increases the risk of hallucinatory-delusional psychosis cerebral atherosclerotic Genesis in 2,98 times, the subject must take preventive measures against cerebral atherosclerosis, which in turn will contribute to preventing the development of hallucinatory-delusional forms of psychosis complicating the disease.

The proposed method is modern and allows a high degree of accuracy to predict the development of hallucinatory-delusional forms of psychosis cerebral atherosclerotic Genesis as in patients with cerebral atherosclerosis, in the early stages of the cerebral and the of erosclerosis, and healthy people.

A method for predicting the development of hallucinatory-delusional forms of psychosis cerebral atherosclerotic Genesis, characterized in that secrete DNA from lymphocytes of peripheral venous blood by polymerase chain reaction of DNA synthesis, amplified fragments locus hSERT gene vector serotonin and identifying genotype 12/10 predict the risk of hallucinatory-delusional forms of psychosis cerebral atherosclerotic Genesis in the subject.