ТRANSPLANTATION OF AUTOLOGOUS CELLS IN COMPLEX OF TREATMENT OF COMPLICATED ARTERIAL FORM IN THE THORACIC OUTLET SYNDROME.

Impact of the autologous cells transplantation in complex of treatment of complicated arterial form of thoracic outlet syndrome was estimated. In accordance to the proce' dure proposed 18 patients were operated on, in 16 patients a standard decompressive and reconstructive operative interventions were performed. The proposed procedure application have promoted improvement of the patients' treatment results due to opti' mization of microcirculation and angiogenesis.

[THE TREATMENT OF PATIENTS FOR ULCERATIVE-NECROTIC DEFECTS IN CHRONIC ISCHEMIA OF THE LOWER EXTREMITY TISSUES, USING BIOTECHNOLOGICAL METHODS].

The results of treatment of 53 patients, suffering "unreconstructable" affection of the lower extremities (LE) arteries and ulcerative-necrotic defects in their distal parts tissues, were analyzed. In 33 patients (the main group) the allotted mononuclear cells fraction, the bone marrow and the plasm, enhanced by thrombocytes, were applied, using multiple injections along a perimeter and into the bottom of the ulcerative-trophic defect in combination with autotransplantation of the bone marrow aspirate into the ischemized tissues--the shin muscles; in 20 patients (the comparison group)--the autotransplantation of the bone marrow aspirate was accomplished into the ischemized tissues--the shin and the foot muscles. In terms up to 3 mo postoperatively a clinical improvement in the main group was noted in 25 (75.7%) patients, the bearing function of the LE was preserved in 90.9%; and in the comparison group--accordingly, in 8 (40%) and 70%. In 36 mo in the main group the bearing function of the LE was preserved in 75.7% patients, and in the comparison group--in 50%. Application of the biotechnological methods proposed in patients, suffering ulcerative-necrotic defects in chronic ischemia of the LE tissues, have promoted the angiogenesis processes activation, the tissues reparation and regeneration, the wounds healing, and the disease clinical course optimization.

Novel controlled attenuation parameter for noninvasive assessment of steatosis using Fibroscan(®): validation in chronic hepatitis C.

A novel controlled attenuation parameter (CAP) has been developed for Fibroscan(®) to assess liver steatosis, simultaneously with liver stiffness measurement (LSM). We assessed CAP diagnostic accuracy in a large cohort of patients with chronic hepatitis C (CHC) virus. A total of 615 patients with CHC, who underwent both Fibroscan(®) and liver biopsy, were analysed. Fibrosis was graded using METAVIR score. Steatosis was categorized by visual assessment as S(0) : steatosis in <10% of hepatocytes, S(1) : 11-33%, S(2) : 34-66% and S(3) : 67-100%. Performances of CAP and liver stiffness were determined using receiver operating characteristic (ROC) curve analysis and cross-validated using the bootstrap method. The Obuchowski measure was used to assess overall accuracy of CAP and to differentiate between steatosis grades. In multivariate analysis, CAP was related to steatosis (P < 10(-15) ) independently of fibrosis stage (which was related to LSM). The areas under ROC curves using CAP to detect steatosis were 0.80 (95% CI, 0.75-0.84) for S ≥ S(1) , 0.86 (0.81-0.92) for S ≥ S(2) and 0.88 (0.73-1) S = S(3) . CAP exhibited a good ability to differentiate steatosis grades (Obuchowski measure = 0.92). Performance of LSM for fibrosis assessment confirmed results from previous studies. CAP is a novel tool to assess the degree of steatosis and both fibrosis and steatosis can be evaluated noninvasively during the same procedure using Fibroscan(®) , in patients with CHC.

[Alternative method of surgical treatment for aortic arteritis in non-repairable forms of vessels damage].

The purpose of the work--the choice of optimal treatment tactics of patients at the extremity tissue ischemia due to nonspecific aortoarteritis (NA). In the surgical treatment of patients with NA, particularly with concomitant peripheral lesions, and to a greater degree of terminal arterial bed, significantly limited the possibility of direct revascularization. Conservative treatment and subthoracal sympathectomy are ineffective. Autotransplantation of mesenchymal stem cells (MSCs), adipose tissue or bone marrow in ischemic tissue in the form of aspirates or selected from these aspirates cell cultures allowed to correct hemodynamic disorders by stimulating angiogenesis and development of collateral circulation.

[Application of autologous aspirates as well as multipotent stromal cells of the bone marrow and adipose tissue in vascular surgery].

Basing on experiments, performed on laboratory animals, and on the literature data, there was proved stimulating influence of transplantation of stromal cells, residing in adipose tissue, in a kind of stromal-adiposal fraction towards angiogenesis and reparative processes in tissues in ischemic conditions. There was elaborated and introduced the tactics of treatment in patients, suffering chronic ischemia of the lower extremities tissues, using combined methods of surgical treatment, including the bone marrow aspirate (BMA) autotransplantation into the ischemized tissues, the rotation ostheotrepanation, reconstructive and endovascular interventions on proximal segment of the extremity. The efficacy and potential of application of the BMA and adipose tissue was proved in the treatment of patients, suffering ischemia of the upper extremities tissues, caused by thromboobliterating affection of peripheral vascular bed.

Congenital absence of vasopressin and age-dependent changes in ACTH and corticosterone stress responses in rats.

The hypothalamic components of the hypothalamo-pituitary-adrenal axis (HPA) are corticotropin-releasing hormone (CRH) and vasopressin. To test the hypothesis that HPA regulation changes with age, we compared ether and bacterial lipopolysaccharide (LPS) injection induced stress reactions in adult and 10-day-old Brattleboro rats, which naturally lack vasopressin owing to mutation of the gene (di/di). The LPS stimulus was used also with V(1b) receptor antagonist pretreatment (SSR149415). In adult di/di or V(1b) pretreated rats, we observed normal pituitary and adrenocortical secretory responses, while in all 10-day-old rats stress-induced serum corticosterone increases were marked, but adrenocorticotropin (ACTH) increases were significantly smaller. Compared to control pups the adenohypophysis of the 10-day-old di/di rats responded normally to CRH, but their adrenal glands were hyper-responsive to ACTH, while in adults there was greater secretion at both levels with no difference between the genotypes. The serum transcortin level was higher in adults than pups, with the di/di pups having higher transcortin levels than controls. Hence, using the same stressors in adults and pups with both a genetic model and pharmacological pretreatment, we have shown that the role of vasopressin in ACTH regulation is more important during the neonatal period than in adulthood. Blunted hypophysial sensitivity to CRH and similar adrenal gland sensitivity to ACTH in the pups compared to adults suggest that hypothalamic factors could be responsible for the neonatal stress hyporesponsive period.

[The upper extremities revascularization in patients with brachial artery thrombosis due to its chronic damaging by crutches: new approaches to the treatment].

An optimal tactics of treatment of patients, suffering the upper extremities (UE) ischemia, caused by arterial thrombosis, occurred due to subclavian artery damage in those patients, who apply crutches for a long period of time, was elaborated. In modern medicine in patients with the UE arterial bed thrombosis with affection of its peripheral, and more in terminal, portion the possibilities of direct revascularization performance are significantly restricted, especially in patients, suffering from constant mechanical injury of armpit and its vascular content, caused by crutches. Application of the method, consisting of automyelotransplantation of the bone marrow aspirate, have permitted to regulate the disorders of microhemodynamics, using stimulation of angiogenesis and development of a collateral blood flow.

The analgesic effect of pamidronate is not caused by the elevation of beta endorphin level in Paget's disease--a controlled pilot study.

INTRODUCTION: Although an analgesic effect is an essential component of the mode of action of bisphosphonates, its physiological mechanisms are still unclear. Beta-endorphin release plays an important role in the analgesic effect of both calcitonin and raloxifene. As patients with Paget's disease receive large doses of bisphosphonates within relatively short time periods, we examined whether repeated pamidronate infusion therapy would cause measurable change in beta-endorphin levels MATERIALS & METHODS: Visual analog scale (VAS) scores of pain intensity, beta-endorphin levels, and alkaline phosphatase activity of 11 patients with Paget's disease (7 with the mono- and 4 with the polyostotic form) were determined at baseline, as well as after 3 and 6 infusions (on Days 6 and 12 of treatment, respectively). Eleven untreated patients with Paget's disease (7 with the mono- and 4 with the polyostotic form) served as controls. RESULTS: It was established that in the course of pamidronate infusion therapy BE levels remained constant, whereas the values in serum alkaline phosphatase and pain intensity scores were significantly reduced. CONCLUSIONS: Although high-dose pamidronate therapy does mitigate pain substantially (as demonstrated by the reduction of VAS scores), its analgesic action is probably unrelated to the enhancement of beta-endorphin release.

Oestrogen and vitamin D receptor (VDR) genotypes and the expression of ErbB-2 and EGF receptor in human rectal cancers.

Oestrogen/oestrogen receptor (ER) and vitamin D/vitamin D receptor (VDR) systems have been implicated in the pathogenesis of colorectal cancers. The expression of erbB-2 and epidermal growth factor receptor (EGFR) in colorectal cancers has been suggested to have diagnostic and prognostic significance. In our study, XbaI and PvuII polymorphisms of the ER gene and the BsmI polymorphism of the VDR gene were studied in 56 Caucasian patients with rectal cancer. The relationship between the ER and VDR genotypes and the expression of oncogenes was also investigated. The presence of the x allele of ER gene significantly correlated with the overexpression of the erbB-2 and EGFR oncogenes. Significantly increased erbB-2 expression was observed in patients with the VDR B allele. The XXbb allelic combination of the ER/VDR genes was associated with a significantly lower erbB-2 expression, whereas in the other genotypes significantly higher oncogene expression was seen. Our data raise the possibility that ER/VDR gene polymorphisms accompanied by variable oncogene expression might influence the pathogenetic processes of colorectal cancers.

Hormonal and metabolic responses during psychosocial stimulation in aggressive and nonaggressive rats.

The physiological effects of a psychosocial threat (the mere presence of a potentially antagonist individual in the home cage) were studied in aggressive and nonaggressive rats. Aggressive animals spent a significantly longer time with the investigation of the opponent compared with the nonaggressive group. An increase in plasma epinephrine and corticosterone was noticed both in aggressive and nonaggressive animals. Ir beta-endorphin increased significantly only in nonfighters. Glycemia was slightly larger in nonaggressives, while lactaemia increased in both groups. The possibility is discussed that differences in psychosocial stress response may be involved in the regulation of behavior in a real encounter.

Alpha 2-adrenoreceptor subtypes regulate ACTH and beta-endorphin secretions during stress in the rat.

The effect of different alpha 2-adrenoreceptor subtype agonists and antagonists on adrenocorticotrop hormone (ACTH) and beta-endorphin release induced by ether stress was examined. Ether inhalation-induced ACTH and beta-endorphin increase was inhibited by i.c.v. administration of 30 micrograms but not 1 and 10 micrograms clonidine (alpha 2-adrenoreceptor agonist). I.c.v. oxymetazoline (alpha 2A-adrenoreceptor agonist; 1-10-30 micrograms) or the alpha 1-agonist methoxamine (100 micrograms/rat) failed to inhibit the stress-induced rise. Pretreatment with the alpha 1/alpha 2B.C-antagonist prazosin (0.5 mg/kg, i.p.) prevented the effect of clonidine on the ether stress, while the alpha 1/alpha 2A-antagonist WB-4101 (0.5 mg/kg, i.p.) was unable to counteract the inhibitory effect of clonidine. Prazosin alone had no effect on the ether-induced plasma ACTH and beta-endorphin elevation. These results suggest that noradrenaline in the central nervous system may inhibit the stress-induced hypothalamo-pituitary-axis and pituitary beta-endorphin activation via alpha 2B.C-adrenoceptor subtypes and prazosin may antagonize its effect on these receptors.

Catecholaminergic control of intracellular free calcium and beta-endorphin secretion of rat pituitary intermediate lobe cells.

Individual melanotropes and intermediate lobes were tested to elucidate the role of alpha- and beta-adrenergic and D-2 dopamine receptors in the regulation of concentration of intracellular free calcium ([Ca2+]i) and release of beta-endorphin. Hormone secretion was studied in a superfusion system, while [Ca2+]i was measured microspectrofluorimetrically. Noradrenaline (1 microM) resulted in a slight decrease, then a marked increase in [Ca2+]i and secretion of beta-endorphin. The nonselective beta-adrenergic agonist isoproterenol (1 microM) increased [Ca2+]i and secretion of beta-endorphin; this effect was blocked by the beta-antagonist propranolol (10 microM). The alpha-adrenergic agonist phenylephrine (1 microM) increased [Ca2+]i and beta-endorphin secretion, but this effect was not blocked by terazosin or prazosin (alpha1-adrenergic antagonists, 1 microM). Administration of the alpha2-adrenergic agonist xylazine (1 microM) increased [Ca2+]i but did not affect secretion of the hormone. Biphasic effect of noradrenaline was tested in presence of adrenergic and dopaminergic antagonists. The noradrenaline-induced rise in [Ca2+]i and beta-endorphin secretion was decreased by propranolol, but this drug did not modify the inhibition. In the presence of 1 microM sulpiride (selective D-2 dopaminergic antagonist), the inhibitory phase of the curve was abolished, and the subsequent increase was reduced. This suggests that activation of dopamine D-2 receptors is involved not only in the inhibition, but also in the subsequent increase, which may originate from a rebound after the termination of the activation of these inhibitory receptors. Our data suggest the presence of several distinct types of catecholamine receptors in the rat intermediate lobe, and the dominant involvement of D-2 and beta-adrenergic receptors in the noradrenaline-induced regulation of melanotropes.

Compression of the pituitary stalk elicits chronic increases in CSF vasopressin, oxytocin as well as in social investigation and aggressiveness.

The neurochemical and behavioural effects of a novel stereotaxic surgical method developed for interrupting the nerve fibres running through the rat pituitary stalk to the posterior pituitary gland was studied. The cerebrospinal fluid (CSF) vasopressin (AVP) and oxytocin (OT) content as well as changes in aggressiveness were measured in rats one week and one month after the surgical intervention. The main results are as follows: (1) the compression of the pituitary stalk elicits a chronic increase in water consumption, as well as in CSF vasopressin and oxytocin content; (2) the surgical intervention increased the frequency of clinch fighting after one week. The increase in aggressiveness accentuated after one month and, in addition, operated animals showed reduced scores of resting while exploratory and social behaviours increased; (3) there was a strong positive correlation between water consumption, vasopressin, and aggressiveness; (4) oxytocin changes showed a positive correlation with variation in social behaviour. The surgical intervention may serve as a model for lesions of the pituitary stalk and formation of ectopic neurohypophyses in humans.

Relationship between blood pressure variability and serum dehydroepiandrosterone sulfate levels.

Decreased diurnal blood pressure variability and low dehydroepiandrosterone sulfate (DHEAS) levels are important predictors of cardiovascular morbidity and mortality. The aim of the study was to determine the relationship between DHEAS levels and diurnal blood pressure variability in normotensive subjects and in patients with essential hypertension of both genders. An ambulatory blood pressure monitor (ABPM), Meditech O2 device and radioimmunoassay were used for ambulatory blood pressure monitoring and the determination of DHEAS levels, respectively. A close correlation (P < .001) was found between the diurnal indices and plasma DHEAS levels of the 387 subjects (86 normotensive and 301 hypertensive patients) participating in the study. Decreased plasma DHEAS levels were associated in both genders, and in both normotensive and hypertensive patients with significantly (P < .001) lower diurnal indices. There was a close correlation (P < .001) between the age-related decrease in plasma DHEAS levels and diurnal indices in both genders. Systolic and diastolic blood pressure variability changed parallel to plasma DHEAS levels in both genders, whether hypertension was present or not. Additional investigations are needed to find out whether reduced DHEAS levels play a role in decreased diurnal indices or whether both can be traced back to one and the same cause.

Alpha 2-adrenoceptor blockade, pituitary-adrenal hormones, and agonistic interactions in rats.

The effects of adrenergic activation on aggressiveness and the aggression induced endocrine changes were tested in rats. Alpha 2 adrenoceptor blockers were used for enhancing activation of the adrenergic system, and changes in aggressiveness were tested in resident-intruder contests. Three experiments were conducted. In experiment 1, saline injected rats responded to the presence of an opponent by aggression and the increase in plasma ACTH and corticosterone. Intraperitoneal administration of 1 mg/kg CH-38083 (an alpha 2 adrenoceptor antagonist) produced a several fold increase in clinch fighting and mutual upright scores, and also further enhanced the plasma ACTH and corticosterone response. In experiment 2, the effect of three doses (0.5, 1 and 2 mg/kg) of three different alpha 2 adrenoceptor blockers CH-38083, idazoxan and yohimbine were tested. All the substances increased aggression at 0.5 and 1 mg/kg; at 2 mg/kg the effect of idazoxan and yohimbine disappeared, while with CH-38083 an additional increase was obtained. In yohimbine treated animals the enhancement of aggression was reduced already at 1 mg/kg. In experiment 3, indomethacin, a potent inhibitor of the catecholamine-induced ACTH release completely abolished the effects of the alpha 2 adrenoceptor antagonist CH-38083: the intensity of agonistic interactions, as well as ACTH and corticosterone plasma concentrations, returned to control levels. The possible role of catecholamines and the stress hormones in the activation of aggression is discussed.

Beta endorphin-like immunoreactivity in human aqueous humor and crystalline lens.

We measured the concentration of beta-endorphin (beta-End) in plasma, as well as in aqueous humor and crystalline lens removed during cataract surgery. beta-End was detected both in the aqueous humor and in the crystalline lens. The concentration of beta-End in the aqueous humor corresponded to almost the half of the plasma level (2.18 fmol/l and 4.55 fmol/l). Endogenous beta-End is presumed to enter the intraocular structures by passive diffusion.

Neonatal monosodium glutamate treatment abolishes both delta opioid receptor-induced and alpha-2 adrenoceptor-mediated gastroprotection in the lower brainstem in rats.

Neonatal monosodium glutamate treatment reduced immunoreactive beta-endorphin content in the mediobasal hypothalamus by 50% in adult, male Wistar rats as compared to hypertonic saline-treated littermates; there was also a moderate (approx. 25%) reduction in the rostral part of the nucleus of the solitary tract. In sham-treated adults the intracisternally injected alpha-2 adenoceptor stimulant clonidine (0.47 nmol/rat) and the delta opioid receptor type agonist (D-Ala(2), D-Leu(5))-enkephalin (0.8 nmol/rat) reduced acidified ethanol-induced mucosal lesions in the stomach by 84.1 and 77.5%, respectively, whereas the same doses were completely ineffective in rats treated neonatally by monosodium glutamate. The data taken together with the results of previous studies with the same substances in rats with retroarcuate knife cuts suggest that neuronal damage in the nucleus of the solitary tract region rather than in the arcuate nucleus is responsible for the changes seen in the pharmacological responsiveness.

A serine-proteinase inhibitor (Boc-D-Phe-Pro-Arg-H) inhibits the secretion of adrenocorticotropin- and beta-endorphin-immunoreactive peptides in vitro.

The effects of a potent serine protease inhibitor, Boc-D-Phe-Pro-Arg-H, on the secretion and content of ACTH and beta-endorphin-like immunoreactivities of cultured rat anterior pituitary cells were studied. Basal release of the hormones was not affected by the drug but secretion in response to stalk-median eminence extracts was inhibited. The inhibitor did not effect the amount of ACTH or beta-endorphin in the medium plus the cells, nor did it change the ratio of the lipotropin and beta-endorphin-like immunoreactivities. Thus the compound seems to affect the release of ACTH and beta-endorphin, but not the processing of lipotropin to beta-endorphin.

Effects of mediobasal hypothalamic lesion on immunoreactive ACTH/beta-endorphin levels in cerebrospinal fluid, in discrete brain regions, in plasma, and in pituitary of the rat.

One week after complete destruction of the mediobasal hypothalamus, immunoreactive adrenocorticotropin (ACTH) and beta-endorphin levels were determined in cerebrospinal fluid, trunk blood, as well as in brain and pituitary tissue samples collected from anaesthetized and cisternally cannulated rats. Control rats were sham operated. In lesioned rats we observed: (a) 60% decrease in the immunoreactive beta-endorphin concentrations in the cerebrospinal fluid, (b) decreased immunoreactive ACTH and beta-endorphin levels in the hypothalamus, in the thalamus and in the amygdala, (c) unaffected immunoreactive ACTH/beta-endorphin levels in the septum and in the hippocampus, (d) decreased immunoreactive beta-endorphin levels both in the anterior and neurointermediate pituitary but unchanged immunoreactive ACTH contents in the anterior lobe, and (e) unaffected immunoreactive ACTH and beta-endorphin levels in the plasma under stressful conditions. From these findings the following conclusions can be drawn: (1) more than 50% of the beta-endorphin-like peptide content of the cerebrospinal fluid originates from the periventricular nuclei of the hypothalamus and thalamus in the rat; (2) the loss of the hypothalamic control probably enhances the intracellular proteolytic degradation of beta-endorphin both in the anterior and neurointermediate pituitary lobe; (3) rats with mediobasal hypothalamic lesion cannot react to the stressful stimuli of ether anaesthesia or cisternal cannulation with elevated plasma immunoreactive ACTH and beta-endorphin levels.