Blood platelet and "extra-platelet" serotonin in some myeloproliferative disorders

Most of the blood serotonin is bound to platelets. However, some authors described plasma "free" serotonin; which is normally very low, and might be increased during some diseases. Usual spectrofluorimetric methods and paired by statistical evaluation "extra-platelet" serotonin (not bound to platelets) in the blood of healthy volunteers and patients with myeloproliferative disorders was studied. In these conditions, we cannot be sure of the existence of "extra-platelets" serotonin in the blood of controls and some patients; in others it was present. "Extra-platelet" serotonin is always present in the blood of untreated patients with primary thrombocytosis.     

The interhemispheric brain asymmetry of the rabbit in a state of "animal hypnosis"]

By the method of forced immobilization the rabbits were brought into the state of "animal hypnosis" (immobilization reflex), and their ECoG was recorded, which was further processed on the computer. It was found that during hypnosis a functional interhemispheric brain asymmetry was developed in rabbits with activity predominance in the right hemisphere. The "animal hypnosis" is a phasic process: in the ECoG of the rabbit under hypnosis a regular alternation of delta and theta activity takes place. Electrophysiological reconstructions in the rabbit brain during the change of its functional state correlate with the brain thermal reactions, revealed earlier.     

Cryptosporidium spp. in domestic dogs: the "dog" genotype

Genetic and phylogenetic characterization of Cryptosporidium isolates at two loci (18S rRNA gene and heat shock gene) from both Australian and United States dogs demonstrated that dog-derived Cryptosporidium isolates had a distinct genotype which is conserved across geographic areas. Phylogenetic analysis provided support for the idea that the "dog" genotype is, in fact, a valid species.     

"Ischemia/reperfusion" for stem cells of two "critical" cell renewal systems of organism

In this article is presented the result of the experiments on mice-hybrids F1(CBA x C57B1/6), which indicates the presence of the reaction of "ischemia/reperfusion" for stem cells of two "critical" cell renewal systems of organism (bone marrow and intestinal epithelium) during the irradiation under the conditions of hypoxic radioprotector application. The additional injection of the source of nitric oxide radicals-sodum nitroprusside (SNT) to the mice right after the irradiation under the conditions of hypoxic protection by serotonin, resulted the substantial increase of the survival rate of hematopoietic stem cells (registered by the methods of endogenous and exogenous colony forming in spleen) and stem cells of intestinal epithelium (registered by the method of intestinal "microcolonies"). The similar radioprotective effect was also registered during the test of survival rate of mice under tests of "bone marrow" and "intestinal" forms of radiation lethality that is evidence of the importance of the realization of phenomenon "ischemia/reperfusion" in the reaction of whole organism on the acute radiation injury. As SNP weakens the manifestation apoptosis and necrosis through competition with active forms of oxygen (AFO) during the period of "reperfusion" on basis of the found out phenomenon experimental model for studying mechanisms of stem cells damage in vivo induced by AFO and for the search of the modifiers weakening or strengthening such damage can be developed.     

Serotonin; its possible relation to allergic disease

Increased amounts of serotonin as well as histamine have been found in the blood of animals during anaphylactic shock. Certain animals, particularly those in which antihistamines do not prevent anaphylaxis, have been found to have increased quantities of serotonin in the lung tissue during anaphylactic shock. Serotonin is a chemical derived from the amino acid tryptophan, which is widely distributed. It is excreted in the urine as the metabolite 5-hydroxyindoleacetic acid. Serotonin has been found in increased amounts in the blood of patients with carcinoids. The increase of serotonin in the blood and the finding of the excretory product in the urine has become a corroborative sign of the disease. The involvement of serotonin in the production of mental disease is evidenced by the effect of serotonin antagonists, which appear to influence mental behavior.That serotonin antagonists may be of ultimate value in the treatment of allergic disease is a possibility to be considered.     

SEROTONIN—Its Possible Relation to Allergic Disease

Increased amounts of serotonin as well as histamine have been found in the blood of animals during anaphylactic shock. Certain animals, particularly those in which antihistamines do not prevent anaphylaxis, have been found to have increased quantities of serotonin in the lung tissue during anaphylactic shock.Serotonin is a chemical derived from the amino acid tryptophan, which is widely distributed. It is excreted in the urine as the metabolite 5-hydroxyindoleacetic acid. Serotonin has been found in increased amounts in the blood of patients with carcinoids. The increase of serotonin in the blood and the finding of the excretory product in the urine has become a corroborative sign of the disease. The involvement of serotonin in the production of mental disease is evidenced by the effect of serotonin antagonists, which appear to influence mental behavior.That serotonin antagonists may be of ultimate value in the treatment of allergic disease is a possibility to be considered.     

Characteristics of the spatio-temporal organization of the P300 component of AEP during the "active" and "passive" stimulus perception in healthy subjects

The concern of the work was in detection and analysis of P300 component of the acoustic evoked potential in healthy subjects in different experimental situations. During counting the rare sounds, P300 was most pronounced in the frontocentral and parietooccipital areas mainly of the left-hemisphere. The response shape was correlated with characteristics of the basic rhythm of the background EEG. Responses of simple and complex shapes were distinguished. The simplest responses were recorded in subjects with hypersynchronous alpha-rhythm. Analysis of three-dimensional dipole source localization showed that structures of the brainstem, limbic system, and frontal lobes participate in generation of the wave. In all the subjects, the decisive role in response generation was played by the brainstem structures. In persons with hypersynchronous alpha-rhythm, the contribution of the frontal lobes was less pronounced. During "passive" listening of sounds, P300 parameters significantly differed from those observed during counting only in 46% of cases (in persons having no hypersynchronous alpha-rhythm). A simplification of the response shape during "passive" listening was observed in these cases, the area of the maximal response expression was shifted to symmetrical areas of the right-hemisphere, the number of dipole sources reduced due to a decrease in the contribution of the frontal and limbic structures into the response generation.     

Plasmodium berghei: a mouse model for the "sudden death" and "malarial lung" syndromes

A mouse model for the "sudden death" and "malarial lung" syndromes is described. Mice of the C3H/z strain succumb suddenly approximately 7 days after an infection with Plasmodium berghei becomes patent, at a time when parasitemia is still moderate (6 to 8%). Death could be shown to be due to anaphylactoid shock, probably induced by soluble immune complexes. Increased vascular permeability caused transudation and leakage of serum proteins into the interstitium and the alveoli. The lungs were found to be edematous, with a fine granular precipitate in the alveoli and adherent to the vascular walls. The precipitates reacted with antiglobulins G and M, and could be shown to also contain malaria antigens and C3/4. A dramatic drop in hematocrit was recorded several hours before death, indicating the sudden release of malaria antigens. The myocardium of animals that had died very suddenly showed a patchy loss of phosphorylase activity. This loss of activity was much more extensive, and sometimes almost total, when there had been an agonal period of several (1 to 3) hours before death. In these cases the irreversibility of the myocardial damage was also indicated by the loss of activity of the dehydrogenases, as well as by typical inflammatory reactions of granulocytic and histiocytic infiltrations. The hearts thus presented a typical picture of the acute and peracute shock syndromes. In acute shock cardiac insufficiency develops so suddenly that death ensues before irreversible damage has occurred, and cardiac insufficiency can only be demonstrated by the most sensitive of enzyme histochemical means. In the present case shock was induced by the anaphylactoid activity of immune complexes with the lung as target organ. The described syndrome appears analogous to human "malarial lung."     

Traumatic shock causes elevation of the serotonin 5-HT2B receptor mRNA level in rat aorta

Previously we have shown that at traumatic shock in rats the force of contraction of isolated aorta in response to angiotensin II, vasopressin, endothelin 1, or norepinephrine is decreased. On the contrary, vasoconstriction caused by serotonin is increased. A possible reason of the alterations of neuroendocrine regulation of vascular tone in shock may be a change in the expression of the receptors of these agonists in blood vessels. In the present study, using real-time PCR, we demonstrated that a day after injury the contents of mRNA encoding receptors V1A for vasopressin, ETA for endothelin 1, and AT1 for angiotensin II are not changed in aorta. There was a slight increase of the serotonin 5-HT2A receptor mRNA (36 ± 16%; p = 0.41). The level of the 5-HT2B receptor mRNA in aorta, initially low (2% of the content of the mRNA of receptors 5-HT2A), after the injury increased 15.8 ± 0.3 times (p < 0.01). However, at traumatic shock there was no contraction of aorta in response to 5-HT2B receptor agonist BW723C86, while vasodilation of the isolated aorta preconstricted with norepinephrine in response to BW723C86 was similar to that of the vessel isolated from control rats. The data obtained suggest that the observed 5-HT2B receptor overexpression is not related to the increased serotonin-induced vasoconstriction and might cause other vascular pathological changes at traumatic shock.     

Catecholamines in Regulation of Development of GnRH Neurons of Rat Fetuses

The contents of dopamine, serotonin, and noradrenaline in rat fetuses developing under the conditions of their deficiency induced by administration of α-methyl-para-tyrosine to females during 11th to 16th or 20th day of pregnancy and in fetuses, whose mothers were given saline at the same time, were determined using HPLC with subsequent electrochemical detection. Administration of α-methyl-para-tyrosine led to decreased levels of dopamine and noradrenaline in the areas of migration of GnRH-neurons in fetuses on days 17 and 21 of prenatal development. The concentration of serotonin remained unchanged, except in the head nasal area in males on day 21. The areas of interaction between the brain catecholaminergic systems and migrating and differentiating GnRH-neurons were determined by double immunohistochemical labeling. Close topographical location of GnRH-immunoreactive neurons and tyrosine hydroxylase-immunoreactive in the area of nucleus accumbens on days 17 and 20, as well as in the median eminence on day 20. The GnRH concentration in the caudal areas of migration of GnRH-neurons under the normal conditions and in the case of catecholamine deficiency was determined using radioimmunoassay. After administration of α-methyl-para-tyrosine the GnRH concentration in the anterior hypothalamus decreased in females. The data obtained suggest the involvement of catecholamines in the regulation of development of GnRH-Neurons during prenatal development. In addition, the adequacy and efficiency of the used model of catecholamine deficiency for studying the development of such neurons was confirmed.     

Pharmacological profile of the "triple" monoamine neurotransmitter uptake inhibitor, DOV 102,677

1. The molecular and behavioral pharmacology of DOV 102,677 is characterized. 2. This characterization was performed using radioligand binding and neurotransmitter uptake assays targeting the monoamine neurotransmitter receptors. In addition, the effects of DOV 102,677 on extracellular neurotransmitter levels were investigated using in vivo microdialysis. Finally, the effects of DOV 102,677 in the forced swim test, locomotor function, and response to prepulse inhibition was investigated.3. DOV 102,677 is a novel, "triple" uptake inhibitor that suppresses [(3)H]dopamine (DA), [(3)H]norepinephrine (NE) and [(3)H]serotonin (5-HT) uptake by recombinant human transporters with IC(50) values of 129, 103 and 133 nM, respectively. Radioligand binding to the dopamine (DAT), norepinephrine (NET), and serotonin (SERT) transporters is inhibited with k (i) values of 222, 1030, and 740 nM, respectively. DOV 102,677 (20 mg/kg IP) increased extracellular levels of DA and 5-HT in the prefrontal cortex to 320 and 280% above baseline 100 min after administration. DA levels were stably increased for the duration (240 min) of the study, but serotonin levels declined to baseline by 200 min after administration. NE levels increased linearly to a maximum of 348% at 240 min post-dosing. Consistent with these increases in NE levels, the density of beta-adrenoceptors was selectively decreased in the cortex of rats treated with DOV 102,677 (20 mg/kg per day, PO, 35 days). 4. DOV 102,677 dose-dependently reduced the amount of time spent immobile by rats in the forced swim test, a model predictive of antidepressant activity, with a minimum effective dose (MED) of 20 mg/kg and a maximal efficacy comparable to imipramine. This decrease in immobility time did not appear to result from increased motor activity. Further, DOV 102,677 was as effective as methylphenidate in reducing the amplitude of the startle response in juvenile mice, without notably altering motor activity. 5. In summary, DOV 102,677 is an orally active, "balanced" inhibitor of DAT, NET and SERT with therapeutic versatility in treating neuropsychiatric disorders beyond depression.     

Adrenoreceptors are involved in the stimulation of neutrophils by exercise-induced circulating concentrations of Hsp72: cAMP as a potential "intracellular danger signal"

Recently, the terms "stress mediators" or "danger signals" have come to be used to describe endogenous molecules that can be released in stress situations and activate the innate immune system even in the absence of antigenic stimuli. There is evidence suggesting that extracellular heat shock proteins of 72 kDa (eHsp72), together with noradrenaline (NA), are candidates as danger signals during exercise-induced stress, interacting in the activation of neutrophils. Previous studies have shown that the post-exercise circulating concentration of eHsp72 activates the phagocytic process of neutrophils with the participation of toll-like receptor 2, but that other receptors must also be involved. The present investigation evaluates the role of adrenoreceptors in the activation of the chemotaxis, phagocytosis, and fungicidal capacity of neutrophils by the post-exercise circulating concentration of eHsp72. The results showed that intact α- and β-adrenoreceptors are necessary for the stimulation of all stages of the phagocytic process by eHsp72. Also, eHsp72 increased the intracellular levels of cAMP, suggesting that it is an "intracellular danger signal" during stress-induced activation of neutrophils mediated by extracellular heat shock proteins. These results can contribute to better understanding the mechanisms involved in the regulation of the innate immune response mediated by "danger signals" during exercise, and probably during other stress situations.     

Inhibition of a conditioned reflex evoked emotional response as an "instrument" of avoidance

A method has been elaborated of training the human subjects to avoid electric shocks by overcoming the "fear" caused by a warning signal. A previously elaborated differentiated conditioned skin-galvanic response (SGR) to a sound stimulus served as a criterion of the emotional reaction. The SGR was projected on an oscilloscope screen. The electric shock was automatically delivered at the moment when the amplitude of the conditioned SGR exceeded the level marked on the screen. The subject was instructed, by overcoming the "fear", to retain the SGR amplitude below the level. If the subject succeeded in overcoming the conditioned SGR to the signal, the shock was not delivered. Since, the suppression of the conditioned SGR served as a "instrument" of avoidance and became consolidated as a habit based on a positive reinforcement. The subjects developed a stable attraction to repetition of training sessions in which they learned to suppress the "anxiety" caused by a signal of "threat".     

Effect of chronic stress on the ultrastructure of the myocardium and hypothalamus of "emotional" and "nonemotional" rats

The influence of chronic stress on the ultrastructure of the myocardium and hypothalamus was studied in experiments on male rats with different levels of emotional-behavioral reactivity. "Emotional" rats manifested a pronounced increase in glycogen granules in myocytes and intercellular space, appearance of the areas of overcontraction of myofibrils, conglomerates of aggregated platelets in myocardial capillaries, and red cell egress from myocardial and hypothalamic capillaries. Alterations in the ultrastructure of the myocardium and hypothalamus in "nonemotional" rats were less marked and consisted in the appearance of the areas of overcontraction of myofibrils, enlargement of sarcoplasmic reticulum caverns, and in an increase in the lipid content in cardiomyocytes. The ultrastructural changes in the myocardium and hypothalamus of "emotional" and "nonemotional" rats indicate different reactivity of the animals and are likely to be accounted for by different levels of activation of their adrenergic systems.     

Electron microscopic observations of the anterior pituitary gland. Part I. The neurons in the "transitional zone" of the anterior pituitary gland

Since [Westlud, K.N., Chils, G.V., 1982. Localization of serotonin fibers in the rat adenohypophysis. Endocrinology 111, 1761-1763] initially identified the serotonin nerve fibers in the anterior pituitary gland, attention has been paid to the rostral zone of the anterior lobe into which nerve fibers enter and subsequently spread to deeper regions of the lobe. The rostral zone is the trifurcated junction of the partes tuberalis, intermedia and distalis, and has the important role(s) for hormone secretion via the "transitional zone" [Sato, G, Shirasawa, N, Sakuma, E, Sato, Y, Asai, Y, Wada, I, Horiuchi, O, Sakamoto, A, Herbert, DC, Soji, T, 2005a. Intercellular communications within the rat anterior pituitary. XI: An immunohistochemical study of distributions of S-100 positive cells in the anterior pituitary of the rat. Tissue and Cell 37, 269-280.]. The objective of this study was to focus on the ultrastructure of this "zone." All of the animals studied were fixed by perfusion with glutaraldehyde via the left ventricle of the heart and examined by electron microscopy. In the "transitional zone," a cluster of neuronal elements was observed between the folliculo-stellate cell-rich area and the anterior lobe. This cluster consisted of myelinated fibers, unmyelinated fibers, neuroendocrine fibers, large cells, and supporting cells. The large cells were perikarya of neurons which made a "ganglion-like" structure with associated satellite cells. Agranular, folliculo-stellate cells were intermingled among the elements. This is the first report that neuronal elements form clusters in the "transitional zone." A relationship of the unmyelinated and neuroendocrine fibers in the basal layer and in the "transitional zone" is discussed.     

Reaction of the elements of the hypophyseal-adrenal system in "starting" states

Morphological elements of the system hypophysis--adrenals have been studied in 19 dogs. Nine animals were given single individually dosed physical loading (running in the treadmill) up to the III stage according to an original technique. The III stage of the organism's response is characterized, according to physiological and biochemical parameters, as a "starting" condition. The other 10 animals served as a control. Biometric, histologic, histoenzymatic and electron microscopic methods were used for the study. The "starting" conditions were demonstrated to determine variously directed dynamics and unequal involvment of cellular elements and zones of the system hypophysis--adrenals. The greatest reactive degree was noted in basophilic adenocytes and chromophobes, as well as in the medullar substance of the adrenals. Acidophils and cells of the fascicular zone play the least role in the response. Unequal participation of the adenohypophyseal cellular elements and the adrenal zones in the organism's response at the "starting" conditions form a predominance of certain processes of cellular activity in them.     

Variability of bacterial biofilms of the "tina" wood vats used in the ragusano cheese-making process

Ragusano cheese is a "protected denomination of origin" cheese made in the Hyblean region of Sicily from raw milk using traditional wooden tools, without starter. To explore the Ragusano bacterial ecosystem, molecular fingerprinting was conducted at different times during the ripening and biofilms from the wooden vats called "tinas" were investigated. Raw milks collected at two farm sites, one on the mountain and one at sea level, were processed to produce Ragusano cheese. Raw milk, curd before and after cooking, curd at stretching time (cheese 0 time), and cheese samples (4 and 7 months) were analyzed by PCR-temporal temperature gel electrophoresis (PCR-TTGE) and by classical enumeration microbiology. With the use of universal primers, PCR-TTGE revealed many differences between the raw milk profiles, but also notable common bands identified as Streptococcus thermophilus, Lactobacillus lactis, Lactobacillus delbrueckii, and Enterococcus faecium. After the stretching, TTGE profiles revealed three to five dominant species only through the entire process of ripening. In the biofilms of the two tinas used, one to five species were detected, S. thermophilus being predominant in both. Biofilms from five other tinas were also analyzed by PCR-TTGE, PCR-denaturating gradient gel electrophoresis, specific PCR tests, and sequencing, confirming the predominance of lactic acid bacteria (S. thermophilus, L. lactis, and L. delbrueckii subsp. lactis) and the presence of a few high-GC-content species, like coryneform bacteria. The spontaneous acidification of raw milks before and after contact with the five tinas was followed in two independent experiments. The lag period before acidification can be up to 5 h, depending on the raw milk and the specific tina, highlighting the complexity of this natural inoculation system.     

Engineering of <Emphasis Type="Italic">Escherichia coli</Emphasis> for the synthesis of <Emphasis Type="Italic">N</Emphasis>-hydroxycinnamoyl tryptamine and serotonin

Plants synthesize various phenol amides. Among them, hydroxycinnamoyl (HC) tryptamines and serotonins exhibit antioxidant, anti-inflammatory, and anti-atherogenic activities. We synthesized HC–tryptamines and HC–serotonin from several HCs and either tryptamine or serotonin using Escherichia coli harboring the 4CL (4-coumaroyl CoA ligase) and CaHCTT [hydroxycinnamoyl-coenzyme A:serotonin N-(hydroxycinnamoyl)transferase] genes. E. coli was engineered to synthesize N-cinnamoyl tryptamine from glucose. TDC (tryptophan decarboxylase) and PAL (phenylalanine ammonia lyase) along with 4CL and CaHCTT were introduced into E. coli and the phenylalanine biosynthetic pathway of E. coli was engineered. Using this strategy, approximately 110.6 mg/L of N-cinnamoyl tryptamine was synthesized. By feeding 100 μM serotonin into the E. coli culture, which could induce the synthesis of cinnamic acid or p-coumaric acid, more than 99 μM of N-cinnamoyl serotonin and N-(p-coumaroyl) serotonin were synthesized.     

No change in the density of the serotonin1A receptor, the serotonin4 receptor or the serotonin transporter in the dorsolateral prefrontal cortex from subjects with schizophrenia

Changes in serotonin receptors and the serotonin transporter have been reported in the dorsolateral prefrontal cortex from subjects with schizophrenia, an area of the brain thought to be important in the pathology of the illness. To further our understanding on how such changes could play a role in the pathology of the illness, in situ radioligand binding with autoradiography was used to measure the density of the serotonin1A receptor, the serotonin4 receptor and the serotonin transporter in the dorsolateral prefrontal cortex, obtained at autopsy, from 10 schizophrenic and 10 control subjects. The binding of [3H]8-OH-DPAT to serotonin1A receptor, [3H]GR113808 to the 5HT4 receptor and [3H]citalopram to serotonin transporter was not altered in subjects with schizophrenia. significantly, only in tissue from the control subjects was there a relationship between age and the density of the serotonin4 receptor in Brodmann's areas 8 (r = 0.71, P = 0.02) and 10 (r = -0.67, P = 0.03). Importantly, this confounding factor did not influence the comparison of the density of serotonin4 receptor in the tissue from the schizophrenic and control subjects. This study has failed to show a difference in the density of serotonin1A receptor, the serotonin4 receptor or the serotonin transporter in the dorsolateral prefrontal cortex (Brodmann's areas 8, 9 and 10) from subjects with schizophrenia. These data suggest that not all serotonergic markers are altered in the dorsolateral prefrontal cortex from schizophrenic subjects.