Intranasal neuropeptide Y reverses anxiety and depressive-like behavior impaired by single prolonged stress PTSD model.

PTSD is a debilitating neuropsychiatric disorder and many patients do not respond sufficiently to current treatments. Neuropeptide Y (NPY) is suggested to provide resilience to the development of PTSD and co-morbid depression. Injections of NPY to the rodent brain are anxiolytic. Recently we showed that intranasal delivery of NPY to rats before or immediately after exposure to single prolonged stress (SPS) animal model of PTSD prevented development of many biochemical and behavioral symptoms of PTSD, indicating its prophylactic potential. Here, we investigated whether intranasal NPY might provide benefits once symptoms have already developed. One week after exposure to SPS stressors, animals were given intranasal NPY or vehicle and tested on elevated plus maze 2h or 2 days later. The NPY treated rats had lower anxiety-like behavior than vehicle treated rats as indicated by more entries into open arms and fewer into closed arms, lower anxiety index, higher risk assessment and unprotected head dips and reduced grooming time. Their anxiety index was similar to that of unstressed controls. On most of these variables there was no effect of time interval and rats displayed similar overall changes 2h or 2 days after the infusion. Moreover, intranasal NPY led to reduced depressive-like behavior, assessed by forced swim test. Thus, intranasal NPY reversed several behavioral impairments triggered by the traumatic stress of SPS and has potential for non-invasive PTSD therapeutic intervention.

[The association between cerebral atrophy and amounts of alcohol consumption in alcoholic patients].

OBJECTIVE: To study the relationship between alcohol consumption and cerebral atrophic changes. MATERIAL AND METHODS: Forty-four second stage alcoholic patients at the age of 30-55 years were examined using magneto-resonance imaging of the brain combined with psychological testing of cognitive functions and the TLFB technique. Based on the TLFB technique, patients were stratified into mild and heavy alcohol consumers. RESULTS AND CONCLUSION: All patients had signs of atrophy in the cerebral cortex, especially in its fronto-parietal parts, the corpus callosum and paraventricular white matter. The degree of the atrophy and weakening of cognitive functions in the mild alcohol consumers was considerably higher than in the heavy alcohol consumers. This suggests the relatively higher sensitivity to alcohol in such patients that is associated with less favorable outcome.

Single intranasal neuropeptide Y infusion attenuates development of PTSD-like symptoms to traumatic stress in rats.

Exposure to severe stress leads to development of neuropsychiatric disorders, including depression and Post-Traumatic Stress Disorder (PTSD) in at-risk individuals. Neuropeptide Y (NPY) is associated with resilience or improved recovery. Therefore exogenous administration to the brain has therapeutic potential although peripheral administration can trigger undesirable side effects. Here, we established conditions with intranasal (IN) NPY infusion to rats to obtain CSF concentrations in the proposed anxiolytic range without significant change in plasma NPY. Rats were pretreated with IN NPY or vehicle before exposure to single prolonged stress (SPS) animal model of PTSD and compared to untreated controls. The IN NPY appeared to lessen the perceived severity of stress, as these animals displayed less time immobile in forced swim part of the SPS. Thirty minutes after SPS the elevation of plasma adrenocorticotropic hormone (ACTH) and corticosterone was not as pronounced in NPY-infused rats and the induction of tyrosine hydroxylase (TH) in locus coeruleus (LC) was attenuated. Seven days after SPS, they displayed lower depressive-like behavior on Forced Swim Test and reduced anxiety-like behavior on Elevated Plus Maze. The prolonged effect of SPS on Acoustic Startle Response was also lower in NPY-infused rats. Plasma ACTH, corticosterone, and hippocampal glucocorticoid receptor levels were significantly above controls only in the vehicle - but not IN NPY-treated group 1week after SPS. Baseline TH mRNA levels in LC did not differ among groups, but increased with forced swim in the vehicle - but not NPY-pretreated animals. Administration of IN NPY after exposure to SPS led to similar, but not identical, reduction in development of anxiety, depressive-like behavior and hyperarousal. The results show that single IN NPY can alter stress-triggered dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis and activation of central noradrenergic activity. These findings provide proof of concept for potential of IN NPY for non-invasive prophylactic treatment or early intervention in response to traumatic stress.

[Clinical and genetic characteristics of long-livers in Moscow region].

In Moscow region long-livers we have studied distribution of LPL, CETP, APOE, F2, F5, F7, F13, FGB, ITGA2, ITGB3, PAI-1, MTHFR, MTRR, HLA-DRB1, HLA-DQA1, HLA-DQB1 genes polymorphisms, associated with predisposition to age pathology. Long-livers are characterized by favorable course of cardiovascular diseases accompanied by certain genetic factors. We have established that genotype H-H- of LPL, allele epsilon2 of APOE, genotype CC of MTHFR (677C > T), genotype TC of ITGB3, genotype GA of FGB, HLA-DRB1*11 positively correlate with longevity.

[The use of immunomodulating therapy as part of complex treatment of secondary peritonitis induces reduction of inflammation in patients of different age].

We present experience of using anti-thymocytic immunoglobulin (ATG) in complex therapy of patients with extensive secondary peritonitis at the age of 29-83 years. The research was based on 60 patient cases: 29 (48%) of whom were given anti-thymocytic immunoglobulin (ATG) Antilymfolin in post operative period and 31 patient of the control group who did not receive immunomodulating therapy within the complex treatment. The received data clearly demonstrates the effectiveness of anti-thymocytic immunoglobulin (ATG) usage for normalization of innate immunity indices and inflammation reduction. Immunostimulating effect in patients given anti-thymocytic immunoglobulin (ATG) could be seen on the fourth day of the threatment. The drug is equally effective in patients of young and middle age as well as in patients of elderly and senility age. The positive influence of anti-thymocytic immunoglobulin (ATG) on inflammation is shown with the reduction of CRP, gamma-globuline, alpha1-protein fraction serum levels normalization. The use of Antilymfolin induces the regression of inflammation and apparently improves the quality and duration of treatment and rehabilitation.

[Medical and social characteristics of the very old Moscow citizens and World War II veterans].

The very old people is a growing population group in Russia and in Moscow in particular that requires more close attention as it demands special medical and social approaches. The investigation was based on data obtained from Moscow district clinics concerning the gender and pathologies structure of people aged 90 yrs and older. In this group women prevailed in total amount and among those older 100 yrs. Striking gender difference was revealed in disturbances of the carbohydrate metabolism. The most frequent pathologies in this population are cardiovascular, nerve and gastrointestine duct systems diseases that have equal prevalence among men and women. Centenarians appeared to be healthier than those in the group of 90-94 yrs. Many of them could serve themselves without extra help (36,48% versus 13,06%). The results obtained in this research must be helpful in the organization of geriatric clinics and cabinets.

Varied mechanisms of oestradiol-mediated regulation of dopamine ß-hydroxylase transcription.

Experiments performed in vivo and in cell culture have demonstrated that oestradiol induces dopamine ß-hydroxylase (DBH) gene transcription. In the present study, we examined oestrogen-responsive elements of the rat DBH gene promoter aiming to characterise the mechanisms of oestradiol-induced DBH transcription. Various mutations and deletions of DBH promoter reporter constructs were tested for responsiveness to 17ß-oestradiol (E(2) ). Mutation of the half palindromic oestrogen response element (ERE) at position -759 reduced the response to E(2) in PC12 cells co-transfected with oestrogen receptor (ER) α, indicating a functional role for this motif. In cells co-transfected with ERß, mutations at the -759 site were unresponsive to E(2) . To characterise the additional E(2) responsive elements, mediated by ERα, the DBH promoter was truncated to the proximal 249 or 200 nucleotides upstream of the transcription start site. Despite either truncation, 10 nm E(2) still elicited an approximately two-fold induction of DBH promoter activity. Mutation of a possible ERE-like sequence at -59 had no effect. The lack of a functional ERE in the proximal region of the rat DBH promoter despite E(2) -mediated DBH promoter activity, suggests regulation by a nonclassical mechanism, such as a membrane-initiated signalling pathway. Moreover, the induction of DBH promoter activity and the rise in DBH mRNA levels were observed within hours. To determine whether membrane-initiated E(2) signalling is involved in rat DBH gene transcription, a membrane impermeable E(2) conjugate, ß-oestradiol-6-(O-carboxy-methyl) oxime-bovine serum albumin (E(2) BSA), was used. Incubation with E(2) -BSA induced luciferase activity and elicited a significant rise in DBH mRNA levels in the ERα transfected cells. The findings indicate two different mechanisms whereby DBH transcription is regulated by E(2) in the presence of ERα. The results implicate both genomic and membrane-initiated mechanisms, mediated by ERα, in E(2) -induced DBH gene transcription.

Adrenocorticotropic hormone elevates gene expression for catecholamine biosynthesis in rat superior cervical ganglia and locus coeruleus by an adrenal independent mechanism.

Classically, upon hypothalamic stimulation, adrenocorticotropic hormone (ACTH) is released from the pituitary and acts on melanocortin 2 receptors (MC2R) in the adrenal cortex, stimulating glucocorticoid synthesis and release. Our earlier studies suggested that ACTH might have a direct effect on sympathetic ganglia. To analyze further the involvement of ACTH in regulation of gene expression of norepinephrine (NE) biosynthetic enzymes, we examined the effect of bilateral adrenalectomy (ADX) of Sprague-Dawley male rats. Fourteen days post-ADX, as expected, plasma ACTH was elevated, and levels of tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH) and MC2R mRNAs in superior cervical ganglia (SCG), and TH mRNA in locus coeruleus (LC) were increased compared with sham-operated animals. To determine effect of pulsatile elevation of ACTH, corticosterone pellets were implanted to ADX rats. Similar to immobilization (IMO) stress ACTH injections to these animals caused a rise in ACTH in plasma and triggered elevation of TH and DBH mRNAs in SCG and in LC with single and repeated daily injections, and MC2R mRNA in SCG with single injections. To study the effect of ACTH in isolated cells, primary cultures of rat SCG were transfected with TH and DBH promoter constructs and treated with ACTH. In agreement with the in vivo data, ACTH elevated their promoter activities similar to levels triggered by cyclic AMP analog. ACTH in the human SK-N-SH neuroblastoma cells increased TH and DBH promoter activity and endogenous DBH mRNA levels. The results show that ACTH can have a direct effect on transcription and gene expression of NE biosynthetic enzymes even without contribution of adrenal hormones.

Estrogen-triggered activation of GTP cyclohydrolase 1 gene expression: role of estrogen receptor subtypes and interaction with cyclic AMP.

Guanosinetriphosphate cyclohydrolase I (GTPCH) catalyzes the initial step in the de novo biosynthesis of (6R)-5,6,7,8-tetrahydrobiopterin, an important determinant of the rate of catecholamine and nitric oxide biosynthesis. Administration of estrogen in vivo was found to elevate GTPCH mRNA levels in several catecholaminergic locations. To examine the mechanism, PC12 cells were co-transfected with a reporter construct containing 2988 bp of rat GTPCH promoter fused to luciferase gene, and expression vectors for estrogen receptors. Addition of 2.5-20 nM of 17 beta-estradiol increased GTPCH promoter-driven luciferase activity in the presence of either estrogen receptor alpha or estrogen receptor beta indicating, for the first time, that 17 beta-estradiol can regulate GTPCH gene expression via transcriptional mechanisms. However, there were differences in dose dependence and time course with estrogen receptor alpha or estrogen receptor beta. With estrogen receptor alpha, the effect was greater with lower doses of 17 beta-estradiol. At the same dose, the response with estrogen receptor beta was observed somewhat earlier than with estrogen receptor alpha and with 20 nM 17 beta-estradiol was effective even after 6 h. These responses to 17 beta-estradiol required estrogen receptors and specific agonists for estrogen receptor alpha and estrogen receptor beta, 4,4,4,-(4-propil-[1H-pyrazole-1,3,5-triyl)tris-phenol and 2,3-bis[4-hydroxyphenyl]propionitrile respectively, triggered increased GTPCH promoter activity. In addition, neither estradiol, nor the selective agonists activated GTPCH promoter without transfection of appropriate estrogen receptor expression vectors. Addition of 17 beta-estradiol, or the selective agonists, also elevated endogenous GTPCH mRNA levels. The results demonstrate that estrogen can have a direct effect on GTPCH gene expression. Although estradiol increased GTPCH promoter activity in the presence of estrogen receptors, it attenuated the response of the promoter and endogenous gene to cyclic AMP, suggesting the crosstalk between estrogen and cyclic AMP pathways in the regulation of GTPCH gene expression. These findings reveal the significance of estrogen in modulating regulation of rate limiting enzyme in the (6R)-5,6,7,8-tetrahydrobiopterin biosynthesis, which may have implications for sex-related differences in vulnerability in related disorders.

[Reproductive function and viability of progeny in female rats under unfavourable influences in run up to puberty].

Wistar female rats were subjected to a 3.5-day water deprivation once a week in the period of 1.5 to 3 months of age. Their progeny was subjected to the same influences during the same period of life. A week later, reproductive function of female rats was evaluated by mating them with normal males. In the experimental groups of both generations, no significant changes were found in number of neonates and their body mass, in maternal behaviour during the lactation period, in postnatal mortality of pups, in their growth and development, in motor activity, physical endurance and behaviour.

[Adaptation to new environments: microgravity].

Review and analysis of the experiments with Wastar rats in microgravity onboard "Cosmos" biosatellites allows to conclude that adaptive potentials of mammals in space flights lasting up to 1/50 th of their life span are enough for rapid elimination of microgravity-induced metabolic and structural alterations on return to Earth, for maintenance of adequate reactions to acute and chronic stressors in the post-flight period, for normal reproductive function and life span. Consideration is given to individual differences in body responses to the micro-g environment.

Estrogen modifies stress response of catecholamine biosynthetic enzyme genes and cardiovascular system in ovariectomized female rats.

Estrogen is likely involved in the gender specific differences in coping with stress. Activation of catecholamine (CA) biosynthetic enzyme gene expression in central and peripheral CA systems plays a key role in response to stress and in regulation of the cardiovascular system. Here we examined whether estradiol can modulate response of hypothalamic-pituitary-adrenal axis (HPA), gene expression of enzymes related to CA biosynthesis in several noradrenergic locations, tetrahydrobiopterin (BH4) concentration and blood pressure (BP) in response to immobilization stress (IMO) of ovariectomized female rats. Rats were injected with 25 mug/kg estradiol benzoate (EB) or sesame oil once daily for 16 days and subsequently exposed to two hours of IMO. The IMO triggered elevation in plasma ACTH was lessened in EB-pretreated animals. However, estradiol did not alter the IMO-elicited rise of tyrosine hydroxylase mRNA levels in adrenal medulla (AM) and in the nucleus of solitary track (NTS) compared with controls. The response of GTP cyclohydrolase I (GTPCH) mRNA in AM to IMO was also similar in both groups. Several responses to IMO in EB-treated rats were reversed. Instead of IMO-elicited elevation in dopamine beta-hydroxylase mRNA levels in the locus coeruleus, GTPCH mRNA and BH4 levels in the NTS, they were reduced by IMO. In a parallel experiment, BP was monitored during restraint stress. The elevation of BP in response to single or repeated restraint stress was sustained during 2 h in controls and reduced after 70 min stress in EB treated rats. One month after withdrawal of EB treatment, the BP response to restraint was similar to that of rats which never received EB. The results demonstrate that estrogen can modulate responses to stress affecting HPA axis, CA biosynthesis, in central and peripheral noradrenergic systems, and BP.

[Blood levels of micro- and microelements in aged patients suffering from chronic coronal heart disease]. / Makro- i mikroélementy krovi u patsientov pozhilogo i starcheskogo vozrasta, stradaiushchikh khronicheskoi bolezn'iu serdtsa.

The authors measured serum and whole blood levels of macro- and microelements (ME), such as iron, zinc, sulfur, potassium, calcium in 149 aged patients, suffering from coronary heart disease. The levels of the above ME were measured in various age groups: younger than 60, 60 to 74, 80 to 89 and older than 90 years old. In 36 aged women ME levels were established in correlation with bone mineral density.

Ontogenesis of mammals and gravity.

The results of the experiments with Wistar rats in microgravity and 2G hypergravity are summarized. Their analysis allows to conclude that adaptive potentials of adult animals in space flights lasting up to 1/50 of their life span are enough for maintenance of adequate reactions to acute and chronic stressors in the postflight period, rapid elimination of space-induced metabolic and structural alterations on return to Earth, maintenance of normal reproductive function after space flight. In embryological experiments it was demonstrated that during space flight it is possible not only to maintain physiological functions of an adult organism, but to form functions of a developing fetus. The animals that spent the portion of their prenatal development in space flight were capable to go through the entire cycle of postnatal development, up to sexual maturity and reproduction. In ground based centrifuge experiments with 2G it was demonstrated the possibility of realizing, under hypergravity, of all the main stages of prenatal and early postnatal development of rats: fertilization, embryon implantation, fetal development, birth and lactation of progeny. Exposure of rats to microgravity did not reduce their life span post flight. Alterations in biological age of animals were small.

[Growth, reproductive function, and tolerance to load tests in rats under repetitive adverse conditions]. / Rost, reproduktivnaia funktsiia i perenosimost' nagruzochnykh prob u krys pri periodicheski povtoriaiushchikhsia neblagopriiatnykh vozdeistviiakh.

Wistar rats were subjected to a 3.5-day water deprivation once a month in the period of 1 to 10 months of age. The rats' adaptive abilities proved sufficient for compensation of adverse effects, the rats preserved their normal motor activity, emotionality, and orienting behaviour in the intervals between the effects. In the reproductive period, the males manifested a normal sexual behaviour and fertilizing capacity of their spermatozoids. In the end of the experiment, the experimental animals did not differ from the control animals.

[Treatment of purulent-inflammatory complications of polytrauma, diabetes, infected pancreonecrosis]. / Lechenie gnoino-vospalitel'nykh oslozhnenii, soprovazhdaiushchikh politravmu, sakharnyi diabet, infitsirovannyi pankreonekroz.

Results of Antilympholin-K3 use in combined treatment of purulent-inflammatory complications in polytrauma, diabetes, infected pancreonecrosis are analysed. Thirty-nine patients aged from 19 to 74 years were treated. Double injection of Antilympholin-K3 enhanced phagocytic activity of neutrophils, phagocytic index, chemiluminescence of mononuclears and neutrophils, immunoglobulins concentration. Positive dynamics in immunological indices is accompanied by improving clinical status, cleaning of postoperative wounds from pus, improving of epithelisation.

[Microgravity, life span and biological age of animals]. / Nevesomost', prodolzhitel'nost' zhizni i biologicheskii vozrast zhivotnykh.

Summarized are author's and literary data about the microgravity effects on life span and biological age of animals obtained in experiments with laboratory rats flown in biosatellites Kosmos. Exposure of rats in the spaceflight microgravity as long as 3 wk. (up to 1/50th of the life period of this species) did not reduce the life span post flight. Alterations in biological age as judged by the reproductive function, general resistance and tissue regeneration rate were minor and in a number of parameters were significantly less as compared with the shifts resulting from simulation of the physiological effects of microgravity in laboratory (for a similar period). Prospects of investigations into this problem are considered.

[ACE gene I/D-polymorphism and hereditary predisposition to myocardial infarction]. / Sviaz' I/D-poliformizma gena angiotenzinprevrashchaiushchego fermenta s nasledstvennoi predraspolozhennost'iu k infarktu miokarda.

The authors compare distribution of genotype frequencies and alleles of I/D of ACE gene polymorphism in patients with various forms of ischemic heart disease (IHD): with acute myocardial infarction (MI), stable effort angina (functional class II-III); in patients with postinfarction cardiosclerosis (PICS). A relationship was found between I/D polymorphism and acute MI. Frequency of DD genotype in MI patients was 0.57, in controls--0.21, p < 0.0001, RR = 4.9. The DD genotype may serve a marker of hereditary predisposition to MI. Genotype DD frequency in the group with acute MI was higher than that with PICS. In acute MI frequency of allele D was 0.76, in PICS--0.51, p < 0.0005. It is suggested that low frequency of genotype DD in the PICS group results from higher lethality of patients with DD genotype in the nearest rehabilitation period. Patients with repeated MI have a significantly higher frequency of genotype DD and complications after MI. Thus, there is a relationship between insertion-deletion polymorphism of ACE gene and myocardial infarction. Deletion DD genotype raises the risk to develop MI and probability of life-threatening complications and repeated MI.

[HindIII DNA-polymorphism of lipoprotein lipase gene in elderly patients with ischemic heart disease]. / HindIII DNK-polimorfizm gena lipoproteinlipazy u bol'nykh ishemicheskoi bolezn'iu serdtsa pozhilogo vozrasta.

AIM: To determine incidence of HindIII alleles of lipoprotein lipase (LPL) in Russian elderly patients with stable effort angina (SEA) functional class II-III regarding lipid metabolism. MATERIAL AND METHODS: Genotyping by LPL gene was performed in 103 patients with SEA. Of them 13 patients survived myocardial infarction (MI), 29 patients had diabetes mellitus. RESULTS: Incidence of alleles of HindIII DNA-polymorphism of LPL gene both in healthy and IHD patients is comparable with that in the West European populations. CONCLUSION: Genotype H+H+ of LPL gene is one of the markers of predisposition to MI, while allele H- is one of the resistance marks.

Comparison of the reactions of male and female Wistar rats. To 5-day exposure to 2G hypergravity.

Number of essential distinctions between male and female rats both before and after 5-day exposure to 2g were observed. These distinctions are not simple quantitative. One gets an impression that males and females have various "strategies" of homeostasis maintenance in the norm and various patterns of adaptive reactions in different situations including 2g hypergravity.