Sample records for amphetamine-regulated transcript immunoreactivity

  1. Immunohistochemical localization of cocaine- and amphetamine-regulated transcript peptide in the central nervous system of the frog Rana esculenta

    NARCIS (Netherlands)

    Lazar, G.; Calle, M.; Roubos, E.W.; Kozicz, L.T.


    ddThe distribution of cocaine- and amphetamine-regulated transcript peptide (CARTp)-like immunoreactivity was studied only in the rat central nervous system (CNS). In mammals, CART peptides occur among others in brain areas that control feeding behavior. We mapped CARTp-immunoreactive structures in

  2. Cocaine- and amphetamine-regulated transcript is present in hypothalamic neuroendocrine neurones and is released to the hypothalamic-pituitary portal circuit

    DEFF Research Database (Denmark)

    Larsen, P J; Seier, V; Fink-Jensen, A; Holst, Jens Juul; Warberg, J; Vrang, N


    Cocaine- and amphetamine-regulated transcript (CART) is present in a number of hypothalamic nuclei. Besides actions in circuits regulating feeding behaviour and stress responses, the hypothalamic functions of CART are largely unknown. We report that CART immunoreactivity is present in hypothalamic...

  3. Genetic variants in the cocaine- and amphetamine-regulated transcript gene (CARTPT) and cocaine dependence


    Lohoff, Falk W.; Bloch, Paul J.; Weller, Andrew E.; Nall, Aleksandra H.; Doyle, Glenn A.; Buono, Russell J.; Ferraro, Thomas N.; Kampman, Kyle M.; Pettinati, Helen M.; Dackis, Charles A.; Oslin, David W.; O'Brien, Charles P.; BERRETTINI, WADE H.


    Dopaminergic brain systems have been implicated to play a major role in drug reward, thus making genes involved in these circuits plausible candidates for susceptibility to substance use disorders. The cocaine- and amphetamine-regulated transcript peptide (CARTPT) is involved in reward and feeding behavior and has functional characteristics of an endogenous psychostimulant. In this study we tested the hypothesis that variation in the CARTPT gene increases susceptibility to cocaine dependence ...

  4. Temperature dependent changes in cocaine- and amphetamine regulated transcript (CART) peptide in the brain of tadpole, Sylvirana temporalis. (United States)

    Shewale, Swapnil A; Gaupale, Tekchand C; Bhargava, Shobha


    Cocaine- and amphetamine-regulated transcript peptide (CARTp) has emerged as a novel neurotransmitter in the brain. Although the physiological role of the peptide has been intensely investigated in mammals, its role in amphibians has not been investigated. In the present study, an attempt has been undertaken to study the expression of CART in the tadpole brain of frog Sylvirana temporalis, subjected to thermal stress. Cells with strong CART-immunoreactivity were observed in the nucleus preoptic area (NPO) of tadpoles exposed to high temperature (37±2°C) as compared to those in the tadpoles exposed to low (12±2°C) and normal (24±2°C) temperatures. In the ventromedial thalamic nucleus (VM) and nucleus posterocentralis thalami (NPC), moderate CART-ir cells were observed in the control groups while number of cells and intensity of immunoreactivity was increased in tadpoles at low and high temperatures. In the nucleus infundibularis ventralis (NIV) and raphe nucleus (RA), CART immunoreactivity increased in the low as well as high temperature treated groups. Intensely stained CART cells were observed in the pituitary of tadpoles exposed to high temperature as compared to low temperature and control groups. We suggest that CART system in the brain and pituitary of tadpole may play a very important role in mediating responses to temperature variations in the environment. PMID:24983774

  5. Association of Cocaine- and Amphetamine-Regulated Transcript (CART) Messenger RNA Level, Food Intake, and Growth in Channel Catfish (United States)

    Cocaine-and Amphetamine-Regulated Transcript (CART) is a potent hypothalamic anorectic peptide in mammals and fish. We hypothesized that increased food intake is associated with changes in expression of CART mRNA within the brain of channel catfish. Objectives were to clone the CART gene, examine ...

  6. Colocalization of cocaine- and amphetamine-regulated transcript with kisspeptin and neurokinin B in the human infundibular region.

    Directory of Open Access Journals (Sweden)

    Katalin Skrapits

    Full Text Available Kisspeptin (KP- and neurokinin B (NKB- synthesizing neurons of the hypothalamic arcuate nucleus play a pivotal role in the regulation of pulsatile gonadotropin-releasing hormone (GnRH secretion. Unlike in rodents and sheep, the homologous KP and NKB neurons in the human infundibular region rarely express dynorphin- but often exhibit Substance P (SP immunoreactivity, indicating remarkable species differences in the neurochemical phenotype of these neurons. In search for additional neuropeptides in human KP and NKB neurons, we carried out immunofluorescent studies on hypothalamic sections obtained from five postmenopausal women. Colocalization experiments provided evidence for the presence of cocaine- and amphetamine-regulated transcript (CART in 47.9 ± 6.6% of KP-immunoreactive (IR and 30.0 ± 4.9% of NKB-IR perikarya and in 17.0 ± 2.3% of KP-IR and 6.2 ± 2.0% of NKB-IR axon varicosities. All three neuropeptides were present in 33.3 ± 4.9% of KP-IR and 28.2 ± 4.6% of NKB-IR somata, respectively, whereas triple-labeling showed lower incidences in KP-IR (14.3 ± 1.8% and NKB-IR (5.9 ± 2.0% axon varicosities. CART-IR KP and NKB neurons established contacts with other peptidergic cells, including GnRH-IR neurons and also sent projections to the infundibular stalk. KP and NKB fibers with CART often contained SP as well, while being distinct from CART fibers co-containing the orexigenic peptide agouti-related protein. Presence of CART in human, but not rodent, KP and NKB neurons represents a new example of species differences in the neuropeptide repertoire of mediobasal hypothalamic KP and NKB neurons. Target cells, receptor sites and physiological significance of CART in the efferent communication of KP and NKB neurons in primates require clarification.

  7. Role of cocaine- and amphetamine-regulated transcript in estradiol-mediated neuroprotection (United States)

    Xu, Yun; Zhang, Wenri; Klaus, Judith; Young, Jennifer; Koerner, Ines; Sheldahl, Laird C.; Hurn, Patricia D.; Martínez-Murillo, Francisco; Alkayed, Nabil J.


    Estrogen reduces brain injury after experimental cerebral ischemia in part through a genomic mechanism of action. Using DNA microarrays, we analyzed the genomic response of the brain to estradiol, and we identified a transcript, cocaine- and amphetamine-regulated transcript (CART), that is highly induced in the cerebral cortex by estradiol under ischemic conditions. Using in vitro and in vivo models of neural injury, we confirmed and characterized CART mRNA and protein up-regulation by estradiol in surviving neurons, and we demonstrated that i.v. administration of a rat CART peptide is protective against ischemic brain injury in vivo. We further demonstrated binding of cAMP response element (CRE)-binding protein to a CART promoter CRE site in ischemic brain and rapid activation by CART of ERK in primary cultured cortical neurons. The findings suggest that CART is an important player in estrogen-mediated neuroprotection and a potential therapeutic agent for stroke and other neurodegenerative diseases. ischemia | stroke | estrogen

  8. Cocaine- and amphetamine-regulated transcript (CART) protects beta cells against glucotoxicity and increases cell proliferation. (United States)

    Sathanoori, Ramasri; Olde, Björn; Erlinge, David; Göransson, Olga; Wierup, Nils


    Cocaine- and amphetamine-regulated transcript (CART) is an islet peptide that promotes glucose-stimulated insulin secretion in beta cells via cAMP/PKA-dependent pathways. In addition, CART is a regulator of neuronal survival. In this study, we examined the effect of exogenous CART 55-102 on beta cell viability and dissected its signaling mechanisms. Evaluation of DNA fragmentation and chromatin condensation revealed that CART 55-102 reduced glucotoxicity-induced apoptosis in both INS-1 (832/13) cells and isolated rat islets. Glucotoxicity in INS-1 (832/13) cells also caused a 50% reduction of endogenous CART protein. We show that CART increased proliferation in INS-1 (832/13) cells, an effect that was blocked by PKA, PKB, and MEK1 inhibitors. In addition, CART induced phosphorylation of CREB, IRS, PKB, FoxO1, p44/42 MAPK, and p90RSK in INS-1 (832/13) cells and isolated rat islets, all key mediators of cell survival and proliferation. Thus, we demonstrate that CART 55-102 protects beta cells against glucotoxicity and promotes proliferation. Taken together our data point to the potential use of CART in therapeutic interventions targeted at enhancing functional beta cell mass and long-term insulin secretion in T2D. PMID:23250745

  9. Cocaine- and amphetamine-regulated transcript peptide (CART) in the brain of zebra finch, Taeniopygia guttata: Organization, interaction with neuropeptide Y, and response to changes in energy status. (United States)

    Singh, Omprakash; Kumar, Santosh; Singh, Uday; Kumar, Vinod; Lechan, Ronald M; Singru, Praful S


    Cocaine- and amphetamine-regulated transcript (CART) has emerged as a potent anorectic agent. CART is widely distributed in the brain of mammals, amphibians, and teleosts, but the relevant information in avian brain is not available. In birds, CART inhibits food intake, whereas neuropeptide Y (NPY), a well-known orexigenic peptide, stimulates it. How these neuropeptides interact in the brain to regulate energy balance is not known. We studied the distribution of CART-immunoreactivity in the brain of zebra finch, Taeniopygia guttata, its interaction with NPY, and their response to dynamic energy states. CART-immunoreactive fibers were found in the subpallium, hypothalamus, midbrain, and brainstem. Conspicuous CART-immunoreactive cells were observed in the bed nucleus of the stria terminalis, hypothalamic paraventricular, supraoptic, dorsomedial, infundibular (IN), lateral hypothalamic, Edinger-Westphal, and parabrachial nuclei. Hypothalamic sections of fed, fasted, and refed animals were immunostained with cFos, NPY, and CART antisera. Fasting dramatically increased cFos- and NPY-immunoreactivity in the IN, followed by rapid reduction by 2 hours and restoration to normal fed levels 6-10 hours after refeeding. CART-immunoreactive fibers in IN showed a significant reduction during fasting and upregulation with refeeding. Within the IN, double immunofluorescence revealed that 94 ± 2.1% of NPY-immunoreactive neurons were contacted by CART-immunoreactive fibers and 96 ± 2.8% NPY-immunoreactive neurons expressed cFos during fasting. Compared to controls, superfused hypothalamic slices of fasted birds treated with CART-peptide showed a significant reduction (P brain of T. guttata may perform several functions, and has a particularly important role in the hypothalamic regulation of energy homeostasis. J. Comp. Neurol. 524:3014-3041, 2016. © 2016 Wiley Periodicals, Inc. PMID:27018984

  10. Nucleus accumbens cocaine-amphetamine regulated transcript mediates food intake during novelty conflict. (United States)

    Burghardt, P R; Krolewski, D M; Dykhuis, K E; Ching, J; Pinawin, A M; Britton, S L; Koch, L G; Watson, S J; Akil, H


    Obesity is a persistent and pervasive problem, particularly in industrialized nations. It has come to be appreciated that the metabolic health of an individual can influence brain function and subsequent behavioral patterns. To examine the relationship between metabolic phenotype and central systems that regulate behavior, we tested rats with divergent metabolic phenotypes (Low Capacity Runner: LCR vs. High Capacity Runner: HCR) for behavioral responses to the conflict between hunger and environmental novelty using the novelty suppressed feeding (NSF) paradigm. Additionally, we measured expression of mRNA, for peptides involved in energy management, in response to fasting. Following a 24-h fast, LCR rats showed lower latencies to begin eating in a novel environment compared to HCR rats. A 48-h fast equilibrated the latency to begin eating in the novel environment. A 24-h fast differentially affected expression of cocaine-amphetamine regulated transcript (CART) mRNA in the nucleus accumbens (NAc), where 24-h of fasting reduced CART mRNA in LCR rats. Bilateral microinjections of CART 55-102 peptide into the NAc increased the latency to begin eating in the NSF paradigm following a 24-h fast in LCR rats. These results indicate that metabolic phenotype influences how animals cope with the conflict between hunger and novelty, and that these differences are at least partially mediated by CART signaling in the NAc. For individuals with poor metabolic health who have to navigate food-rich and stressful environments, changes in central systems that mediate conflicting drives may feed into the rates of obesity and exacerbate the difficulty individuals have in maintaining weight loss. PMID:26926827

  11. Nicotine regulates cocaine-amphetamine-Regulated Transcript (Cart) in the mesocorticolimbic system. (United States)

    Kaya, Egemen; Gozen, Oguz; Ugur, Muzeyyen; Koylu, Ersin O; Kanit, Lutfiye; Balkan, Burcu


    Cocaine-and-Amphetamine Regulated Transcript (CART) mRNA and peptides are intensely expressed in the brain regions comprising mesocorticolimbic system. Studies suggest that CART peptides may have a role in the regulation of reward circuitry. The present study aimed to examine the effect of nicotine on CART expression in the mesocorticolimbic system. Three different doses of nicotine (0.2, 0.4, 0.6 mg/kg free base) were injected subcutaneously for 5 days, and on day 6, rats were decapitated following a challenge dose. CART mRNA and peptide levels in medial prefrontal cortex (mPFC), nucleus accumbens (NAc), dorsal striatum (DST), amygdala (AMG), lateral hypothalamic area (LHA), and ventral tegmental area (VTA) were measured by quantitative real-time PCR (qPCR) and Western Blot analysis, respectively. In the mPFC, 0.4 and 0.6 mg/kg nicotine, decreased CART peptide levels whereas there was no effect on CART mRNA levels. In the VTA, a down-regulation of CART peptide expression was observed with 0.2 and 0.6 mg/kg nicotine. Conversely, 0.4 and 0.6 mg/kg nicotine increased CART mRNA levels in the AMG without affecting the CART peptide expression. Nicotine did not regulate CART mRNA or CART peptide expression in the NAc, DST, and LHA. We conclude that nicotine regulates CART expression in the mesocorticolimbic system and this regulation may play an important role in nicotine reward. Synapse 70:283-292, 2016. © 2016 Wiley Periodicals, Inc. PMID:26990424

  12. Maternal and fetal cocaine- and amphetamine-regulated transcript in diabetic and non-diabetic pregnancy.

    LENUS (Irish Health Repository)

    Hehir, Mark P


    Cocaine- and amphetamine-regulated transcript (CART) is a leptin-regulated anorectic neuropeptide. Increased levels of leptin in cord blood of diabetic mothers have previously been described. The aim of this study was to quantify maternal and fetal serum CART levels in type 1 diabetes mellitus (T1DM, n = 10) and non-diabetic pregnancy (n = 10). Matched maternal serum samples (n = 20) were obtained at 36-weeks gestation and cord samples from the umbilical vein at delivery (n = 20), CART was quantified using a competitive enzyme immunoassay. Statistical analysis was performed using Spearmans correlation and t test. There was no difference in maternal CART levels at 36-weeks gestation between T1DM (mean = 331.13 pg\\/ml, Standard Error of the Mean (SEM) = 114.54) and non-diabetic pregnancy (mean = 195.01 pg\\/ml SEM = 29.37) (p = 0.106). Fetal CART levels in the umbilical vein were similar in T1DM (mean = 199.27 pg\\/ml, SEM = 39.81) and non-diabetic pregnancy (mean = 149.76 pg\\/ml, SEM = 26.08) (p = 0.143). Maternal serum CART levels measured at 36-weeks gestation correlated with maternal BMI at booking (Spearmans ρ = 0.332) (p = 0.001) irrespective of diabetes. Serum CART can be detected in both diabetic and non-diabetic human pregnancy and may play an important role in body mass regulation in pregnancy.

  13. Evidence for the participation of cocaine- and amphetamine-regulated transcript peptide (CART) in the fluoxetine-induced anti-hyperalgesia in neuropathic rats. (United States)

    Upadhya, Manoj A; Dandekar, Manoj P; Kokare, Dadasaheb M; Singru, Praful S; Subhedar, Nishikant K


    Cocaine- and amphetamine-regulated transcript peptide (CART) has a role in chronic pain, and also in the actions of selective serotonin reuptake inhibitors (SSRIs) employed in the treatment of neuropathic pain. Herein, we test the hypothesis that CART may mediate the anti-hyperalgesic effect of the SSRI, fluoxetine, in neuropathic rats. Sciatic nerve in the right hind paw of rat was ligated to induce neuropathic pain, and the paw withdrawal latency was evaluated using Hargreaves apparatus. Fluoxetine [5-25mg/kg, intraperitoneal (ip)] or CART (54-102) [0.1-1.5μg/rat, intracerebroventricular (icv)] dose-dependently attenuated the hyperalgesic response observed in neuropathic rats, indicating anti-nociceptive properties of each agent. The anti-hyperalgesic effect of fluoxetine was potentiated by the subeffective dose of CART, and attenuated by CART-antibody (1:500 dilution; 5μl/rat, icv); CART-antibody had no effect per se. Isobolographic analysis showed a significant synergism between fluoxetine and CART, and antagonism between fluoxetine and CART-antibody. Immunocytochemical labeling with monoclonal antibodies against CART showed drastic increase in CART-immunoreactive fibers in the ventrolateral periaqueductal gray (VLPAG; 116%), dorsal subdivision of dorsal raphe nucleus (DRD; 176%), and locus coeruleus (LC; 733%) of neuropathic animals. Fluoxetine treatment significantly reduced the immunoreactivity in these areas. However, CART-immunoreactive cells and fibers in the arcuate nucleus did not respond to neuropathy or fluoxetine treatments. We suggest that the CART innervation of DRD, LC and VLPAG may be involved in the (i) central processing of neuropathic pain and (ii) fluoxetine-induced anti-hyperalgesic effect in neuropathic pain. PMID:21167239

  14. Intrafollicular expression and potential regulatory role of cocaine- and amphetamine-regulated transcript in the ovine ovary. (United States)

    Huang, Y; Yao, X L; Meng, J Z; Liu, Y; Jiang, X L; Chen, J W; Li, P F; Ren, Y S; Liu, W Z; Yao, J B; Folger, J K; Smith, G W; Lv, L H


    Follicular growth is regulated by a complex interaction of pituitary gonadotropins with local regulatory molecules. Previous studies demonstrated an important role for cocaine- and amphetamine-regulated transcript (CART) in regulation of granulosa cell estradiol production associated with dominant follicle selection in cattle. However, intraovarian expression and actions of CART in other species, including sheep, are not known. The objective of this study was to investigate the expression of CART in sheep follicles and determine the effects of CART on indices of ovine granulosa cell function linked to follicular development. Results demonstrated the expression of CART messenger RNA and prominent intraovarian localization of CART peptide in granulosa cells of sheep follicles. Granulosa cell CART messenger RNA was lower, but follicular fluid estradiol concentrations were higher in large (>5 mm) follicles vs smaller 3- to 5-mm follicles harvested from sheep ovaries of abattoir origin. CART treatment inhibited follicle stimulating hormone-induced estradiol production by cultured ovine granulosal cells and also blocked the follicle stimulating hormone-induced increase in granulosa cell numbers. Results demonstrate expression of CART in sheep follicular tissues and suggest potential biological actions of CART, which are inhibitory to ovine follicular growth and development. PMID:26490113

  15. The polymorphisms of bovine cocaine- and amphetamine-regulated transcripts and their associations with cattle (Bos taurus) growth traits

    Indian Academy of Sciences (India)

    Chun Lei Zhang; Hong Chen; Yan Hong Wang; Xian Yong Lan; Chu Zhao Lei; Xing Tang Fang


    We investigated the polymorphisms of bovine cocaine- and amphetamine-regulated transcripts (CART). The coding and regulating regions of CART were screened in 7 cattle breeds by the single-stranded conformation polymorphism (SSCP) technique. The four loci (C1, C2, C3 and C4) studied were all polymorphic. Polymerase chain reaction (PCR) products representing different SSCP variants were sequenced and a total of 9 single-nucleotide polymorphisms (SNPs) were found. The associations between polymorphic loci and the growth traits of Nanyang cattle were analysed. The results indicated that genotype A1A1 of the C1 locus was associated with a higher body weight ( < 0.05) than heterozygous A1B1. Genotype A2A2 of the C2 locus was associated with lower body weight and average daily weight gain ( ≤ 0.001) than heterozygous A2B2. C3 and C4 loci had no significant effect on Nanyang cattle growth traits (P > 0.05).

  16. A cocaine-regulated and amphetamine-regulated transcript inhibits oxidative stress in neurons deprived of oxygen and glucose. (United States)

    Sha, Dujuan; Wang, Zhongyuan; Qian, Lai; Han, Yong; Zhang, Jun; Gu, Shuangshuang; Wang, Luna; Li, Jie; Chen, Cong; Xu, Yun


    Stroke, of which about 87% is ischemic stroke, constitutes one of the main causes of morbidity, disability, and mortality worldwide. Ischemic brain injury has complex pathological mechanisms. Considerable evidence has been collected over the last few years suggesting that oxidative stress associated with excessive production of reactive oxygen species is a fundamental mechanism of brain damage in stroke and reperfusion after stroke. Oxidative stress is an important trigger of neuronal apoptosis in ischemic stroke. In this current study, it was found that cocaine-regulated and amphetamine-regulated transcript 55-102 (CART55-102) inhibited oxygen-induced and glucose deprivation (OGD)-induced neurotoxicity in a dose-dependent manner. The peak dose of CART55-102 was 0.4 nmol/l. In addition, the level of intracellular reactive oxygen species was decreased in OGD-treated neurons in the presence of 0.4 nmol/l CART55-102. Mitochondrial membrane potential (ΔΨm) and mtDNA mRNA expressions were increased in OGD-treated neurons in the presence of 0.4 nmol/l CART55-102. The current study suggests that CART55-102, by inhibiting oxidative stress, may be developed into therapeutic agents for ischemic stroke. PMID:23884173

  17. Cocaine- and amphetamine-regulated transcript: a novel regulator of energy homeostasis expressed in a subpopulation of pancreatic islet cells. (United States)

    Gilon, Patrick


    Type 2 diabetes is characterised by chronic hyperglycaemia and its incidence is highly increased by exaggerated food consumption. It results from a lack of insulin action/production, but growing evidence suggests that it might also involve hyperglucagonaemia and impaired control of glucose homeostasis by the brain. In recent years, the cocaine and amphetamine-regulated transcript (CART) peptides have generated a lot of interest in the battle against obesity because, via the brain, they exert anorexic effects and they increase energy expenditure. They are also localised, outside the brain, in discrete regions of the body and play a hormonal role in controlling various functions. In this issue of Diabetologia, the Wierup group (doi: 10.1007/s00125-016-4020-6 ) shows that CART peptides are expressed heterogeneously in islet cells of various species, including humans, and that their expression is upregulated in diabetes. The authors also shine a spotlight on some interesting effects of CART peptides on islet function, including stimulation of insulin secretion and inhibition of glucagon release. CART peptides would thus be at the centre of a cooperation between the brain and the endocrine pancreas to control glucose homeostasis. Although the mechanisms of action of CART peptides remain enigmatic because no specific receptor for these peptides has so far been discovered, their potential therapeutic use is evident and represents a new challenge for future research. PMID:27421727

  18. Characterization of seven cocaine- and amphetamine-regulated transcripts (CARTs) differentially expressed in the brain and peripheral tissues of Solea senegalensis (Kaup). (United States)

    Bonacic, Kruno; Martínez, Almudena; Martín-Robles, Águeda J; Muñoz-Cueto, José A; Morais, Sofia


    CART (cocaine- and amphetamine-regulated transcript) is a peptide with neurotransmitter and neuroendocrine functions with several key roles, both centrally and peripherally. In mammals there is a single gene that produces two alternatively spliced variants in rat and a single transcript in human but in teleosts multiple genes have been found. In the present study we report the existence of seven transcripts in Senegalese sole and characterize their sequences and phylogenetic relationships, as well as their expression patterns in the brain and peripheral tissues, and in response to feeding. Both cart2a and cart4 showed a ubiquitous expression in the brain, while cart1a, cart1b and cart3a were similarly expressed and had higher transcript levels in the mesencephalon, followed by the diencephalon. On the other hand, cart2b showed a main expression in the olfactory bulbs, and cart3b was predominantly expressed in the spinal cord. The expression profile in peripheral tissues differed substantially between cart's, even between more recently duplicated genes. Collectively, all the tissues examined, except the muscle, express at least one of the different cart's, although the highest transcript levels were found in the brain, gonads (ovary and testis) and, in some cases, eye and kidney. Concerning the feeding response, only brain cart1a, cart2a and cart4 showed a significant postprandial regulation, although future studies are necessary to assess potential confounding effects of stress imposed by the force feeding technique employed. Senegalese sole exhibits the highest number of cart genes reported to date in a vertebrate species. Their differential expression patterns and feeding regulation suggest that multiple cart genes, resulting from at least 3 rounds of whole genome duplication, have been retained in fish genomes through subfunctionalization, or possibly even through neofunctionalization. PMID:26320854

  19. Cocaine- and amphetamine-regulated transcript facilitates the neurite outgrowth in cortical neurons after oxygen and glucose deprivation through PTN-dependent pathway. (United States)

    Wang, Y; Qiu, B; Liu, J; Zhu, Wei-Guo; Zhu, S


    Cocaine- and amphetamine-regulated transcript (CART) is a neuropeptide that plays neuroprotective roles in cerebral ischemia and reperfusion (I/R) injury in animal models or oxygen and glucose deprivation (OGD) in cultured neurons. Recent data suggest that intranasal CART treatment facilitates neuroregeneration in stroke brain. However, little is known about the effects of post-treatment with CART during the neuronal recovery after OGD and reoxygenation in cultured primary cortical neurons. The present study was to investigate the role of CART treated after OGD injury in neurons. Primary mouse cortical neurons were subjected to OGD and then treated with CART. Our data show that post-treatment with CART reduced the neuronal apoptosis caused by OGD injury. In addition, CART repaired OGD-impaired cortical neurons by increasing the expression of growth-associated protein 43 (GAP43), which promotes neurite outgrowth. This effect depends on pleiotrophin (PTN) as siRNA-mediated PTN knockdown totally abolished the increase in CART-stimulated GAP43 protein levels. In summary, our findings demonstrate that CART repairs the neuronal injury after OGD by facilitating neurite outgrowth through PTN-dependent pathway. The role for CART in neurite outgrowth makes it a new potential therapeutic agent for the treatment of neurodegenerative diseases. PMID:25010400

  20. Assessing addiction vulnerability with different rat strains and place preference procedures: the role of the cocaine and amphetamine-regulated transcript. (United States)

    Salas, Elisabet; Bocos, Carlos; Castillo, Carmen Del; Pérez-García, Carmen; Morales, Lidia; Alguacil, Luis F


    Validated biomarkers of addiction vulnerability are unavailable despite their potential value in diagnostics and therapeutics. As cocaine and amphetamine-regulated transcript (CART) peptides can be considered candidates for such biomarkers, we have studied the acute regulation of CART gene expression in the nucleus accumbens of rats with different drug-seeking behaviors. Two subgroups of Sprague-Dawley rats with different persistences of cocaine-induced and morphine-induced place preference showed a similar regulation of CART mRNA irrespective of their behavioral differences: CART gene expression was unaffected by acute cocaine and downregulated by acute morphine to a similar extent in both subgroups. Fischer 344 and Lewis rats, known to exhibit very different drug-seeking behaviors, showed lower basal expression of CART when compared with Sprague-Dawley rats, being almost undetectable in the case of the Lewis strain. Acute morphine downregulated CART in Fischer 344 rats as it did in Sprague-Dawley rats. The results tend to show that CART mRNA regulation by acute morphine or cocaine in the nucleus accumbens does not seem predictive of addiction vulnerability. However, in the particular case of Lewis rats, the pronounced hypoactivity of the CART system could contribute to the high vulnerability of this strain to develop drug-seeking behaviors. PMID:23907376

  1. Leptin-Induced CART (Cocaine- and Amphetamine-Regulated Transcript) Is a Novel Intraovarian Mediator of Obesity-Related Infertility in Females. (United States)

    Ma, Xiaoting; Hayes, Emily; Prizant, Hen; Srivastava, Rajesh K; Hammes, Stephen R; Sen, Aritro


    Obesity is considered detrimental to women's reproductive health. Although most of the attention has been focused on the effects of obesity on hypothalamic function, studies suggest a multifactorial impact. In fact, obesity is associated with reduced fecundity even in women with regular cycles, indicating that there may be local ovarian effects modulating fertility. Here we describe a novel mechanism for leptin actions directly in the ovary that may account for some of the negative effects of obesity on ovarian function. We find that normal cycling, obese, hyperleptinemic mice fed with a high-fat diet are subfertile and ovulate fewer oocytes compared with animals fed with a normal diet. Importantly, we show that leptin induces expression of the neuropeptide cocaine- and amphetamine-regulated transcript (CART) in the granulosa cells (GCs) of ovarian follicles both in vitro and in vivo. CART then negatively affects intracellular cAMP levels, MAPK signaling, and aromatase mRNA expression, which leads to lower estradiol synthesis in GCs and altered ovarian folliculogenesis. Finally, in human samples from patients undergoing in vitro fertilization, we show a significant positive correlation between patient body mass index, CART mRNA expression in GCs, and CART peptide levels in follicular fluid. These observations suggest that, under obese conditions, CART acts as a local mediator of leptin in the ovary to cause ovarian dysfunction and reduced fertility. PMID:26730935


    Directory of Open Access Journals (Sweden)

    Jiann Wei Yeoh


    Full Text Available Recent work has established that the paraventricular thalamus (PVT is a central node in the brain reward-seeking pathway. This role is likely mediated in part through the dense projections to the PVT from hypothalamic peptide transmitter systems such as orexin, and cocaine- and amphetamine-regulated transcript (CART, both of which play key roles in drug-seeking behaviour. Consistent with this proposition, we previously found that inactivation of the PVT or infusions of CART into the PVT suppressed drug-seeking behaviour in an animal model of contingent cocaine self-administration. Despite this work, very few studies have assessed the basic physiological properties of PVT neurons and how these parameters are altered by exposure to drugs such as cocaine. We set out to address these questions by employing an electrophysiological approach to record from anterior PVT (aPVT neurons from cocaine-treated and control animals. First, we determined the excitability of aPVT neurons by injecting a series of depolarizing current steps and characterizing the resulting action potential (AP discharge properties. Second, we investigated the effects of CART on excitatory synaptic inputs to aPVT neurons. We found that the majority of aPVT neurons exhibited tonic firing (TF, and initial bursting (IB consistent with previous studies. However, we also identified PVT neurons that exhibited delayed firing (DF, single spiking (SS and reluctant firing (RF. Interestingly, cocaine exposure shifted the proportion of aPVT neurons that exhibited TF. Further, application of CART suppressed excitatory synaptic drive to PVT. This finding is consistent with our previous behavioural data, which showed that CART signaling in the PVT negatively regulates drug-seeking behaviour. Together, these studies support previous anatomical evidence that the PVT can integrate reward-relevant information and provides a putative mechanism through which drugs of abuse can dysregulate this system in

  3. MCG101-induced cancer anorexia-cachexia features altered expression of hypothalamic Nucb2 and Cartpt and increased plasma levels of cocaine- and amphetamine-regulated transcript peptides. (United States)

    Burgos, Jonathan R; Iresjö, Britt-Marie; Smedh, Ulrika


    The aim of the present study was to explore central and peripheral host responses to an anorexia-cachexia producing tumor. We focused on neuroendocrine anorexigenic signals in the hypothalamus, brainstem, pituitary and from the tumor per se. Expression of mRNA for corticotropin-releasing hormone (CRH), cocaine- and amphetamine-regulated transcript (CART), nesfatin-1, thyrotropin (TSH) and the TSH receptor were explored. In addition, we examined changes in plasma TSH, CART peptides (CARTp) and serum amyloid P component (SAP). C57BL/6 mice were implanted with MCG101 tumors or sham-treated. A sham-implanted, pair‑fed (PF) group was included to delineate between primary tumor and secondary effects from reduced feeding. Food intake and body weight were measured daily. mRNA levels from microdissected mouse brain samples were assayed using qPCR, and plasma levels were determined using ELISA. MCG101 tumors expectedly induced anorexia and loss of body weight. Tumor-bearing (TB) mice exhibited an increase in nesfatin-1 mRNA as well as a decrease in CART mRNA in the paraventricular area (PVN). The CART mRNA response was secondary to reduced caloric intake whereas nesfatin-1 mRNA appeared to be tumor-specifically induced. In the pituitary, CART and TSH mRNA were upregulated in the TB and PF animals compared to the freely fed controls. Plasma levels for CARTp were significantly elevated in TB but not PF mice whereas levels of TSH were unaffected. The plasma CARTp response was correlated to the degree of inflammation represented by SAP. The increase in nesfatin-1 mRNA in the PVN highlights nesfatin-1 as a plausible candidate for causing tumor-induced anorexia. CART mRNA expression in the PVN is likely an adaptation to reduced caloric intake secondary to a cancer anorexia-cachexia syndrome (CACS)‑inducing tumor. The MCG101 tumor did not express CART mRNA, thus the elevation of plasma CARTp is host derived and likely driven by inflammation. PMID:26780979

  4. Pheochromocytoma cell PC12 contain binding sites for cart (cocaine- and amphetamine-regulated transcript)

    Czech Academy of Sciences Publication Activity Database

    Maixnerová, Jana; Blokešová, Darja; Matyšková, Resha; Haugvicová, Renata; Šloncová, Eva; Elbert, Tomáš; Železná, Blanka; Maletínská, Lenka

    Praha: Ústav organické chemie a biochemie AV ČR, 2007 - (Slaninová, J.), s. 72-74. (Collection Symposium Series. 9). ISBN 978-80-86241-28-9. [Biologically Active Peptides /10./. Praha (CZ), 11.04.2007-13.04.2007] Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z50200510; CEZ:AV0Z50520514 Keywords : CART peptides * biological activity * specific binding Subject RIV: CE - Biochemistry

  5. Peptid CART (cocaine- and amphetamine- regulated transcript) v signalizaci buněk PC12

    Czech Academy of Sciences Publication Activity Database

    Nagelová, Veronika; Železná, Blanka; Maletínská, Lenka


    Roč. 108, č. 5 (2014), s. 543. ISSN 0009-2770. [Mezioborové setkání mladých biologů, biochemiků a chemiků /14./. 13.05.2014-16.05.2014, Milovy] R&D Projects: GA ČR GAP303/10/1368 Institutional support: RVO:61388963 Keywords : peptide CART * PC12 * c-Jun * SAPK/JNK Subject RIV: CE - Biochemistry

  6. Cocaine- and amphetamine-regulated transcript (CART) peptide specific binding in pheochromocytoma cells PC12

    Czech Academy of Sciences Publication Activity Database

    Maletínská, Lenka; Maixnerová, Jana; Matyšková, Resha; Haugvicová, Renata; Šloncová, Eva; Elbert, Tomáš; Slaninová, Jiřina; Železná, Blanka


    Roč. 559, 2/3 (2007), s. 109-114. ISSN 0014-2999 R&D Projects: GA ČR GA303/05/0614 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z50520514; CEZ:AV0Z50200510 Keywords : radioligand binding * CART * PC12 cells * food intake Subject RIV: CE - Biochemistry Impact factor: 2.376, year: 2007

  7. Pheochromocytoma cell PC12 contain binding sites for cart (cocaine- and amphetamine-regulated transcript)

    Czech Academy of Sciences Publication Activity Database

    Maixnerová, Jana; Blokešová, Darja; Matyšková, Resha; Haugvicová, Renata; Šloncová, Eva; Elbert, Tomáš; Železná, Blanka; Maletínská, Lenka

    Praha: Ústav organické chemie a biochemie AV ČR, 2007. s. 43. ISBN 978-80-86241-00-5. [Biologically Active Peptides /10./. 11.04.2007-13.04.2007, Praha] Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z50520514 Keywords : CART peptides * biological activity * specific binding Subject RIV: CC - Organic Chemistry

  8. Structure-activity relationship of CART (cocaine- and amphetamine-regulated transcript) peptide fragments

    Czech Academy of Sciences Publication Activity Database

    Maixnerová, Jana; Hlaváček, Jan; Blokešová, Darja; Kowalczyk, W.; Elbert, Tomáš; Šanda, Miloslav; Blechová, Miroslava; Železná, Blanka; Slaninová, Jiřina; Maletínská, Lenka


    Roč. 28, č. 10 (2007), s. 1945-1953. ISSN 0196-9781 R&D Projects: GA ČR GA303/05/0614 Institutional research plan: CEZ:AV0Z40550506 Keywords : CART peptide * fragments * binding * PC12 cells Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.368, year: 2007

  9. Strukturně-aktivní studie fragmentů peptidu "cocaine - and amphetamine regulated transcript"

    Czech Academy of Sciences Publication Activity Database

    Maixnerová, Jana; Blokešová, Darja; Železná, Blanka; Maletínská, Lenka


    Roč. 102, č. 5 (2008), s. 384-384. ISSN 0009-2770. [Mezioborové setkání mladých biologů, biochemiků a chemiků. Konference Sigma-Aldrich /8./. 10.06.2008-13.06.2008, Devět skal - Žďárské vrchy] R&D Projects: GA ČR GA303/05/0614 Institutional research plan: CEZ:AV0Z40550506 Keywords : CART peptide * fragments * binding * PC12 cells Subject RIV: CE - Biochemistry

  10. Synergistic effect of CART (cocaine- and amphetamine-regulated transcript) peptide and cholecystokinin on food intake regulation in lean mice

    Czech Academy of Sciences Publication Activity Database

    Maletínská, Lenka; Maixnerová, Jana; Matyšková, Resha; Haugvicová, Renata; Pirnik, Z.; Kiss, A.; Železná, Blanka


    Roč. 9, č. 101 (2008), s. 1-10. ISSN 1471-2202 R&D Projects: GA ČR GA303/05/0614 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z50200510 Keywords : mice * food intake * CART peptide Subject RIV: CE - Biochemistry Impact factor: 2.850, year: 2008

  11. Synergistic effect of CART (cocaine and amphetamine regulated transcript) peptide and cholecystokinin in lean and diet-induced obese mice

    Czech Academy of Sciences Publication Activity Database

    Maixnerová, Jana; Matyšková, Resha; Blokešová, Darja; Haugvicová, Renata; Železná, Blanka; Maletínská, Lenka

    Paris: -, 2007. [World Congress on Prevention and Therapies against Obesity /2./. 14.06.2007-15.06.2007, Paris] Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z50200510 Keywords : cocaine * cholecystokinin * synergy Subject RIV: CE - Biochemistry

  12. Structure-activity relationship of cocaine- and amphetamine-regulated transcript (CART) by peptide analogs: Importance of disulfide bridges

    Czech Academy of Sciences Publication Activity Database

    Blechová, Miroslava; Nagelová, Veronika; Demianova, Zuzana; Železná, Blanka; Maletínská, Lenka


    Roč. 18, S1 (2012), S89-S90. ISSN 1075-2617. [European Peptide Symposium /32./. 02.09.2012-07.09.2012, Athens] Institutional research plan: CEZ:AV0Z40550506 Keywords : CART * neuropeptides * cell line PC12 * anorexigenic effect Subject RIV: CE - Biochemistry

  13. Feeding-related effects of CART (cocaine- and amphetamine-regulated transcript) peptides and cholecystokinin in mouse obese models

    Czech Academy of Sciences Publication Activity Database

    Maletínská, Lenka; Maixnerová, Jana; Matyšková, Resha; Haugvicová, Renata; Slaninová, Jiřina; Železná, Blanka

    Geneva: Kenes International, 2007 - (Rolka, K.; Rekowski, P.; Silberring, J.), s. 254-255 ISBN 978-965-555-297-3. [European Peptide Symposium /29./. Gdansk (PL), 03.09.2006-08.09.2006] R&D Projects: GA ČR GA303/05/0614 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z50200510; CEZ:AV0Z50520514 Keywords : peptides * CART Subject RIV: CC - Organic Chemistry

  14. Feeding-related effects of cart (cocaine and amphetamine regulated transcript) peptides and cholecystokinin in mouse obese models

    Czech Academy of Sciences Publication Activity Database

    Maletínská, Lenka; Maixnerová, Jana; Toma, Resha Shamas; Haugvicová, Renata; Slaninová, Jiřina; Železná, Blanka


    Roč. 12, Supplement (2006), s. 178. ISSN 1075-2617. [European Peptide Symposium /29./. 03.09.2006-08.09.2006, Gdansk] Institutional research plan: CEZ:AV0Z40550506 Keywords : CART peptides * food intake * mouse obesity * CCK Subject RIV: CC - Organic Chemistry

  15. Celiac disease T-cell epitopes from gamma-gliadins: immunoreactivity depends on the genome of origin, transcript frequency, and flanking protein variation

    Directory of Open Access Journals (Sweden)

    Salentijn Elma MJ


    Full Text Available Abstract Background Celiac disease (CD is caused by an uncontrolled immune response to gluten, a heterogeneous mixture of wheat storage proteins. The CD-toxicity of these proteins and their derived peptides is depending on the presence of specific T-cell epitopes (9-mer peptides; CD epitopes that mediate the stimulation of HLA-DQ2/8 restricted T-cells. Next to the thoroughly characterized major T-cell epitopes derived from the α-gliadin fraction of gluten, γ-gliadin peptides are also known to stimulate T-cells of celiac disease patients. To pinpoint CD-toxic γ-gliadins in hexaploid bread wheat, we examined the variation of T-cell epitopes involved in CD in γ-gliadin transcripts of developing bread wheat grains. Results A detailed analysis of the genetic variation present in γ-gliadin transcripts of bread wheat (T. aestivum, allo-hexaploid, carrying the A, B and D genome, together with genomic γ-gliadin sequences from ancestrally related diploid wheat species, enabled the assignment of sequence variants to one of the three genomic γ-gliadin loci, Gli-A1, Gli-B1 or Gli-D1. Almost half of the γ-gliadin transcripts of bread wheat (49% was assigned to locus Gli-D1. Transcripts from each locus differed in CD epitope content and composition. The Gli-D1 transcripts contained the highest frequency of canonical CD epitope cores (on average 10.1 per transcript followed by the Gli-A1 transcripts (8.6 and the Gli-B1 transcripts (5.4. The natural variants of the major CD epitope from γ-gliadins, DQ2-γ-I, showed variation in their capacity to induce in vitro proliferation of a DQ2-γ-I specific and HLA-DQ2 restricted T-cell clone. Conclusions Evaluating the CD epitopes derived from γ-gliadins in their natural context of flanking protein variation, genome specificity and transcript frequency is a significant step towards accurate quantification of the CD toxicity of bread wheat. This approach can be used to predict relative levels of CD toxicity of

  16. Injection of Cocaine-Amphetamine Regulated Transcript (CART) peptide into the nucleus accumbens does not inhibit caffeine-induced locomotor activity: Implications for CART peptide mechanism. (United States)

    Job, Martin O


    Much evidence suggests that intra-nucleus accumbens (NAc) CART peptide (CART 55-102) injection inhibits locomotor activity (LMA) when there is an increase in the release and activity of dopamine (DA) in the NAc. However, this hypothesis has not been fully tested. One way to examine this is to determine if there is a lack of effect of intra-NAc CART peptide on LMA that does not involve increases in DA release in the NAc. Several studies have suggested that caffeine-induced LMA does not involve extracellular DA release in the NAc core. Therefore, in this study, we have examined the effect of injections of CART peptide (2.5μg) into the NAc core on the locomotor effects of caffeine in male Sprague-Dawley rats. Several LMA relevant doses of caffeine were used (0, 10, 20mg/kg i.p.), and an inverted U response curve was found as expected. We determined, in the same animals, that intra-NAc CART peptide had no effect on caffeine-induced LMA whereas it blunted cocaine-mediated LMA, as shown by other reports. We also extended a previous observation in mice by showing that at a LMA activating dose of caffeine there is no alteration of CART peptide levels in the NAc of rats. Our study supports the hypothesis that the inhibitory effects of CART peptide in the NAc may be exerted only under conditions of increased extracellular DA release and activity in this region. Our results also suggest that intra-NAc CART 55-102 does not generally inhibit increases in LMA due to all drugs, but has a more specific inhibitory effect on dopaminergic neurotransmission. PMID:27168116

  17. Cocaine and amphetamine-regulated transcript (CART) concentration in maternal and cord blood in type 1 diabetic and non diabetic pregnancies at term

    LENUS (Irish Health Repository)

    Hehir, MP


    Institute of Obstetricians & Gynaecologists, RCPI Four Provinces Meeting, Junior Obstetrics & Gynaecology Society Annual Scientific Meeting, Royal Academy of Medicine in Ireland Dublin Maternity Hospitals Reports Meeting Nov 2010

  18. CART (cocaine- and amphetamine-regulated transcript) peptide specific binding sites in PC12 cells have characteristics of CART peptide receptors

    Czech Academy of Sciences Publication Activity Database

    Nagelová, Veronika; Pirnik, Z.; Železná, Blanka; Maletínská, Lenka


    Roč. 1547, Feb 14 (2014), s. 16-24. ISSN 0006-8993 R&D Projects: GA ČR GAP303/10/1368 Institutional support: RVO:61388963 Keywords : CART peptide * PC12 cell * differentiation * binding * signaling * c-Jun Subject RIV: CE - Biochemistry Impact factor: 2.843, year: 2014

  19. Cocaine- and Amphetamine-Regulated Transcript (CART) Messenger RNA Expression is Regulated by Food Intake in the Brain of the Channel Catfish (Ictalurus punctatus) (United States)

    Increased growth is included as a selection criterion at the USDA Catfish Genetics Research Unit, Stoneville, MS, USA. Faster growth of selected channel catfish is typically attributed to their ability to consume more feed. While endocrine and neural factors and mechanisms that regulate feed intak...

  20. Iodination of CART (61-102) (cocaine- and amphetamine-regulated transcript) peptide, separation and characterization of products and evaluation of their biological activity

    Czech Academy of Sciences Publication Activity Database

    Elbert, Tomáš; Maletínská, Lenka; Maixnerová, Jana; Železná, Blanka

    Bad Soden: International Isotope Society, 2007. s. 39. [WorkshopThe Synthesis and Applications of Isotopes and Isotopically Labelled Compounds /14./. 21.06.2007-22.06.2007, Bad Soden] Institutional research plan: CEZ:AV0Z40550506 Keywords : CART(61-102) * 125l-labeled peptide Subject RIV: CC - Organic Chemistry

  1. Anorexigenic effect of cholecystokinin is lost but that of CART (cocaine and amphetamine regulated transcript) peptide is preserved in monosodium glutamate obese mice

    Czech Academy of Sciences Publication Activity Database

    Železná, Blanka; Maixnerová, Jana; Matyšková, Resha; Haugvicová, Renata; Blokešová, Darja; Maletínská, Lenka


    Roč. 58, č. 5 (2009), s. 717-723. ISSN 0862-8408 R&D Projects: GA ČR GA303/05/0614 Grant ostatní: GA ČR(CZ) GA305/06/0427 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z5020903 Keywords : monosodium glutamate (MSG) obesity * neuropeptide Y (NPY) * cholecystokinin Subject RIV: CC - Organic Chemistry Impact factor: 1.430, year: 2009

  2. Immunoreactive trypsin in Shwachman's syndrome.


    Dossetor, J F; Spratt, H C; Rolles, C J; Seem, C P; Heeley, A F


    We studied two infants with Shwachman's syndrome in whom the immunoreactive trypsin concentration was found to be abnormally low. Experience with several hundred assays for immunoreactive trypsin has not shown this low concentration. This finding is probably specific for pancreatic acinar deficiency at this age and strongly suggests Shwachman's syndrome.

  3. Effect of anorexinergic peptides, cholecystokinin (CCK) and cocaine and amphetamine regulated transcript (CART) peptide, on the activity of neurons in hypothalamic structures of C57Bl/6 mice involved in the food intake regulation

    Czech Academy of Sciences Publication Activity Database

    Pirnik, Z.; Maixnerová, Jana; Matyšková, Resha; Koutová, Darja; Železná, Blanka; Maletínská, Lenka; Kiss, A.


    Roč. 31, č. 1 (2010), s. 139-144. ISSN 0196-9781 R&D Projects: GA ČR GA303/05/0614 Institutional research plan: CEZ:AV0Z40550506 Keywords : cholecystokinin * CART * hypocretin * Fos peptide Subject RIV: CE - Biochemistry Impact factor: 2.654, year: 2010

  4. Expression of immunoreactive urocortin in human tissue

    Institute of Scientific and Technical Information of China (English)

    GU Qing; Vicki L Clifton; CUI Ying; HUI Ning; ZHOU Xiao-ning; HE Qian; HAN Qing-feng; SHA Jin-yan; Roger Smith


    To localize where urocortin is expressed in human tissue in an attempt to study its physiological functions. Methods: Expression of immunoreactive urocortin in different human tissue was examined using a specific urocortin antibody and the immunoperoxidase staining method. Results: Immunoreactive urocortin was observed in the anterior pituitary cells, decidual stromal cells, syncytiotrophoblasts, amnion epithelium, the vascular smooth muscles of myometrium, fallopian tube and small intestine. Conclusion: The study indicates that urocortin is expressed in some specific areas of human tissue. The data are consistent with the hypothesis that urocortin is produced locally as an endocrine factor, which may act as a neural regulator and a regulator of local blood flow.

  5. Somatostatin-Immunoreactive Pancreaticoduodenal Neuroendocrine Neoplasms

    DEFF Research Database (Denmark)

    Engelund Luna, Iben; Monrad, Nina; Binderup, Tina;


    OBJECTIVE: Neuroendocrine neoplasms in the pancreas and duodenum with predominant or exclusive immunoreactivity for somatostatin (p-dSOMs) are rare, and knowledge on tumour biology, treatment, survival and prognostic factors is limited. This study aimes to describe clinical, pathological, and...

  6. Human lymphocyte production of immunoreactive thyrotropin.


    Smith, E M; Phan, M.; Kruger, T E; Coppenhaver, D H; Blalock, J E


    Interferon-alpha inducers were previously shown to cause human lymphocyte production of a corticotropin (ACTH)-like peptide. Thyrotropin (TSH) was not produced under these conditions. In contrast, this report shows that a T-cell mitogen (staphylococcal enterotoxin A), which does not induce the ACTH-like peptide, caused human lymphocyte production of an immunoreactive (ir) TSH. Lymphocyte synthesis of the ir TSH was first detectable at 24 hr, peaked at 48 hr, and thereafter declined. NaDodSO4/...

  7. Serum immunoreactive trypsin concentrations in diabetic children.


    Moffat, A.; Marks, V.; Gamble, D. R.


    Serum immunoreactive trypsin (SIT) concentrations measured in 616 children with diabetes of recent onset were low, in both boys and girls, in comparison with reference ranges established in patients with non-diabetic, non-infectious illnesses. The mean SIT concentration was 60% of the mean reference level in children tested within three weeks of the onset of diabetes, and about 40% in patients tested six months after the onset of diabetes. Very low SIT levels were found in about 10% of patien...

  8. Clinical value of serum immunoreactive trypsin concentration


    Ruddell, W S J


    The clinical value of estimation of serum concentrations of immunoreactive trypsin was evaluated by studying 46 healthy controls, 23 controls in hospital, 44 patients with chronic pancreatic disease, and 184 patients with non-pancreatic conditions in which pancreatic disease commonly enters into the differential diagnosis. Serum trypsin concentration had a log normal distribution in the controls, and the calculated normal range was considerably wider than that previously reported. The concent...

  9. Effect of Chitosan Properties on Immunoreactivity


    Sruthi Ravindranathan; Bhanu prasanth Koppolu; Smith, Sean G.; Zaharoff, David A


    Chitosan is a widely investigated biopolymer in drug and gene delivery, tissue engineering and vaccine development. However, the immune response to chitosan is not clearly understood due to contradicting results in literature regarding its immunoreactivity. Thus, in this study, we analyzed effects of various biochemical properties, namely degree of deacetylation (DDA), viscosity/polymer length and endotoxin levels, on immune responses by antigen presenting cells (APCs). Chitosan solutions fro...

  10. FMRFamide-like immunoreactivity in the nervous system of Hydra

    DEFF Research Database (Denmark)

    Grimmelikhuijzen, C J; Dockray, G J; Schot, L P


    FMRFamide-like immunoreactivity has been localized in different parts of the hydra nervous system. Immunoreactivity occurs in nerve perikarya and processes in the ectoderm of the lower peduncle region near the basal disk, in the ectoderm of the hypostome and in the ectoderm of the tentacles. The...... immunoreactive nerve perikarya in the lower peduncle region form ganglion-like structures. Radioimmunoassays of extracts of hydra gave displacement curves parallel to standard FMRFamide and values of at least 8 pmol/gram wet weight of FMRFamide-like immunoreactivity. The immunoreactive material eluted from...

  11. Rett Syndrome Mutant Neural Cells Lacks MeCP2 Immunoreactive Bands. (United States)

    Bueno, Carlos; Tabares-Seisdedos, Rafael; Moraleda, Jose M; Martinez, Salvador


    Dysfunctions of MeCP2 protein lead to various neurological disorders such as Rett syndrome and Autism. The exact functions of MeCP2 protein is still far from clear. At a molecular level, there exist contradictory data. MeCP2 protein is considered a single immunoreactive band around 75 kDa by western-blot analysis but several reports have revealed the existence of multiple MeCP2 immunoreactive bands above and below the level where MeCP2 is expected. MeCP2 immunoreactive bands have been interpreted in different ways. Some researchers suggest that multiple MeCP2 immunoreactive bands are unidentified proteins that cross-react with the MeCP2 antibody or degradation product of MeCP2, while others suggest that MeCP2 post-transcriptional processing generates multiple molecular forms linked to cell signaling, but so far they have not been properly analyzed in relation to Rett syndrome experimental models. The purpose of this study is to advance understanding of multiple MeCP2 immunoreactive bands in control neural cells and p.T158M MeCP2e1 mutant cells. We have generated stable wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Application of N- and C- terminal MeCP2 antibodies, and also, RFP antibody minimized concerns about nonspecific cross-reactivity, since they react with the same antigen at different epitopes. We report the existence of multiple MeCP2 immunoreactive bands in control cells, stable wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Also, MeCP2 immunoreactive bands differences were found between wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Slower migration phosphorylated band around 70kDa disappeared in p.T158M MeCP2e1-RFP mutant expressing cells. These data suggest that threonine 158 could represent an important phosphorylation site potentially involved in protein function. Our results clearly indicate that MeCP2 antibodies have no cross-reactivity with similar epitopes on others proteins, supporting the idea that MeCP2 may

  12. Rett Syndrome Mutant Neural Cells Lacks MeCP2 Immunoreactive Bands.

    Directory of Open Access Journals (Sweden)

    Carlos Bueno

    Full Text Available Dysfunctions of MeCP2 protein lead to various neurological disorders such as Rett syndrome and Autism. The exact functions of MeCP2 protein is still far from clear. At a molecular level, there exist contradictory data. MeCP2 protein is considered a single immunoreactive band around 75 kDa by western-blot analysis but several reports have revealed the existence of multiple MeCP2 immunoreactive bands above and below the level where MeCP2 is expected. MeCP2 immunoreactive bands have been interpreted in different ways. Some researchers suggest that multiple MeCP2 immunoreactive bands are unidentified proteins that cross-react with the MeCP2 antibody or degradation product of MeCP2, while others suggest that MeCP2 post-transcriptional processing generates multiple molecular forms linked to cell signaling, but so far they have not been properly analyzed in relation to Rett syndrome experimental models. The purpose of this study is to advance understanding of multiple MeCP2 immunoreactive bands in control neural cells and p.T158M MeCP2e1 mutant cells. We have generated stable wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Application of N- and C- terminal MeCP2 antibodies, and also, RFP antibody minimized concerns about nonspecific cross-reactivity, since they react with the same antigen at different epitopes. We report the existence of multiple MeCP2 immunoreactive bands in control cells, stable wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Also, MeCP2 immunoreactive bands differences were found between wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Slower migration phosphorylated band around 70kDa disappeared in p.T158M MeCP2e1-RFP mutant expressing cells. These data suggest that threonine 158 could represent an important phosphorylation site potentially involved in protein function. Our results clearly indicate that MeCP2 antibodies have no cross-reactivity with similar epitopes on others proteins, supporting the

  13. Rett Syndrome Mutant Neural Cells Lacks MeCP2 Immunoreactive Bands (United States)

    Bueno, Carlos; Tabares-Seisdedos, Rafael; Moraleda, Jose M.; Martinez, Salvador


    Dysfunctions of MeCP2 protein lead to various neurological disorders such as Rett syndrome and Autism. The exact functions of MeCP2 protein is still far from clear. At a molecular level, there exist contradictory data. MeCP2 protein is considered a single immunoreactive band around 75 kDa by western-blot analysis but several reports have revealed the existence of multiple MeCP2 immunoreactive bands above and below the level where MeCP2 is expected. MeCP2 immunoreactive bands have been interpreted in different ways. Some researchers suggest that multiple MeCP2 immunoreactive bands are unidentified proteins that cross-react with the MeCP2 antibody or degradation product of MeCP2, while others suggest that MeCP2 post-transcriptional processing generates multiple molecular forms linked to cell signaling, but so far they have not been properly analyzed in relation to Rett syndrome experimental models. The purpose of this study is to advance understanding of multiple MeCP2 immunoreactive bands in control neural cells and p.T158M MeCP2e1 mutant cells. We have generated stable wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Application of N- and C- terminal MeCP2 antibodies, and also, RFP antibody minimized concerns about nonspecific cross-reactivity, since they react with the same antigen at different epitopes. We report the existence of multiple MeCP2 immunoreactive bands in control cells, stable wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Also, MeCP2 immunoreactive bands differences were found between wild-type and p.T158M MeCP2e1-RFP mutant expressing cells. Slower migration phosphorylated band around 70kDa disappeared in p.T158M MeCP2e1-RFP mutant expressing cells. These data suggest that threonine 158 could represent an important phosphorylation site potentially involved in protein function. Our results clearly indicate that MeCP2 antibodies have no cross-reactivity with similar epitopes on others proteins, supporting the idea that MeCP2 may

  14. Serum immunoreactive trypsin and pancreatic lipase in primary biliary cirrhosis.


    Fonseca, V.; Epstein, O; Katrak, A; Junglee, D; Mikhailidis, D P; McIntyre, N; Dandona, P


    Immunoreactive trypsin concentration and pancreatic lipase activity were measured in the sera of 33 patients with primary biliary cirrhosis. Immunoreactive trypsin was increased (above the normal range) in 16 (48%) and pancreatic lipase activity in 18 (55%) patients. Both enzymes were increased in 10 (30%) patients. Twenty four patients (73%) had an increase of either one or both enzymes. There was a significant correlation between immunoreactive trypsin and pancreatic lipase activity. This a...

  15. Localization in the gastrointestinal tract of immunoreactive prosomatostatin

    DEFF Research Database (Denmark)

    Holst, J J; Poulsen, Steen Seier


    parts of the small intestine but not in the stomach and the colon. The colon contained very few immunoreactive structures. Immunoreactive nerve cell bodies were found in the submucous plexus of the small intestine. All immunoreactive endocrine cells in the stomach and the duodenum and all immunoreactive...... nerves were stained by all 5 antisera whereas the small intestinal endocrine cells did not stain for the most N-terminal region of prosomatostatin. The results suggest that all gastrointestinal somatostatin is derived from the same precursor molecule, which, however, in the small intestinal endocrine...

  16. Immunoreactive trypsin and neonatalscreening for cystic fibrosis

    International Nuclear Information System (INIS)

    Immunoreactive trypsin (IRT) was measured in dried blood spots from 160.822 five-day-old babies as a part of a regionwide neonatal screening program for cystic fibrosis. A second test was performed for 492 babies in whom blood IRT levels were found greater than 900 μg/l; retesting revealed persistent elevation in 55. Sweat testing confirmed cystic fibrosis in 43 babies, but results were normal in 12. During the course of this study, a total of 51 cystic fibrosis babies were identified: 43 by newborn screening, 6 because they had meconium ileus; so, early diagnosis was achieved in 49 cases out of 51. Two newborn babies did not have elevated IRT and they were missed by the screening test. Our results confirm that elevated blood IRT is characteristic of newborn babies with cystic fibrosis and show that this test has an excellent specificity (99.7%) and a good sensitivity (95%) when used as a neonatal screening test

  17. Mechanism of action of cysteamine in depleting prolactin immunoreactivity

    International Nuclear Information System (INIS)

    The thiol reagent cysteamine (CSH) depletes anterior pituitary cells of immunoreactive PRL both in vivo and in vitro. The authors examined the hypothesis that CSH affects either the solubility or immunoreactivity of PRL through a mechanism involving thiol-disulfide exchange. Adult female rats were treated with either CSH (300 mg/kg, sc) or an equimolar dose of ethanolamine as a control. Anterior pituitary glands were extracted in 0.1 M sodium borate buffer, pH 9.0. Treatment of pituitary extracts with beta-mercaptoethanol (BME) destroys the immunoreactivity of PRL. However, extraction in the presence of reduced glutathione or CSH of pituitaries of rats treated with CSH restores immunoreactive PRL to control levels. Extracts were also subjected to polyacrylamide gel electrophoresis (PAGE). On gels of pituitary extracts of CSH-treated rats, the band that comigrates with purified PRL is diminished compared to that in ethanolamine-treated controls. However, extraction of the pituitaries in sodium dodecyl sulfate-containing buffer followed by chemical reduction with BME restores the PRL band. Therefore, CSH acts on PRL through a thiol-related mechanism to yield a product that is poorly soluble in aqueous buffer at pH 9 and is poorly immunoreactive. Dispersed anterior pituitary cells in tissue culture were incubated with L-[35S]methionine to radiolabel newly synthesized peptides. PAGE followed by autoradiography confirmed the above results obtained in vivo

  18. Capsaicin receptor immunoreactivity in the human trigeminal ganglion

    DEFF Research Database (Denmark)

    Hou, Mingyan; Uddman, Rolf; Tajti, Janos; Kanje, Martin; Edvinsson, Lars


    protons, and therefore it is suggested as a molecular integrator of chemical and physical stimuli that elicit pain. In the present study, indirect immunofluorescence detected a small number of neurons that are VR1 receptor immunoreactive (ir) (171 versus 1038 or 16% of all neuronal cell bodies) in the...

  19. 21 CFR 862.1405 - Immunoreactive insulin test system. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Immunoreactive insulin test system. 862.1405 Section 862.1405 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Chemistry...

  20. Bombesin-like immunoreactivity in the nervous system of hydra

    DEFF Research Database (Denmark)

    Grimmelikhuijzen, C J; Dockray, G J; Yanaihara, N


    With immunocytochemical methods, nerve cells have been detected in Hydra attenuata containing bombesin-like immunoreactivity. These nerve cells are located in ectoderm of all body regions of the animal and are especially abundant in basal disk and tentacles. Radioimmunoassay of extracts of hydra...

  1. Immunoreactivity of 125I-papain labelled by different methods

    International Nuclear Information System (INIS)

    Three different methods of papain iodination (with chloramine-T, lactoperoxidase and conjugation with Bolton-Hunter reagent) have been compared. The highest yield of 125I-papain could be obtained using lactoperoxidase which enabled to achieve the highest immunoreactivity. 125I-papain, labelled this way, is suitable for the radioimmunoassay of papain. (author)


    NARCIS (Netherlands)



    Inhibin immunoreactivity was estimated in a number of gonadal and non-gonadal tumors. Dog Sertoli cell tumors and human granulosa cell and Leydig cell tumors contained high concentrations of inhibin-like material. Levels, comparable with those in normal testes and ovaries were detected in human test

  3. γ2-MSH immunoreactivity in the human heart

    International Nuclear Information System (INIS)

    In patients undergoing aorto-coronary by-pass surgery, we found a 26% arterial-venous difference of immunoreactive γ2-melanocytostimulating hormone (MSH), a proopiomelanocortin (POMC) derived peptide known to possess profound hemodynamic effects. These results prompted an investigation of the presence of γ2-MSH in the human heart. Using a two-step extraction procedure, regions of human hearts were examined by sensitive and specific radioimmunoassays to determine their γ2-MSH content. Mean (± SEM) concentrations of 0.14 ± 0.023 pmol/g and 0.12 ± 0.017 were found in right atrium and right ventricle, respectively. High performance liquid chromatography indicated that 80-90 % of the total immunoreactivity eluted in a single sharp peak in a position identical to that of synthetic γ2-MSH

  4. Insulin-like immunoreactive substances in the rat

    International Nuclear Information System (INIS)

    Chromatography on G50 or G100 sephadex column of rat plasma or serum divides up the insulin-like immunoreactive material into three peaks: monomere insulin, proinsulin and a fraction of molecular weight between 50 and 100,000. This fraction is virtually absent (less than 1%) from immunoreactive material extracted from the pancreas. Comparison of the results obtained by methods using double or simple antibodies (charcoal dextran) and study of fixation in vitro of labelled insulin, taken up by various plasma proteins, suggest that the high molecular weight material includes insulin more or less broken down and linked to proteins. Furthermore, when a double antibody method is used, the alpha globulins and albumin in the rat present also an insulin-like reactivity. This disadvantage does not occur with the charcoal dextran method which is more specific

  5. gamma. sub 2 -MSH immunoreactivity in the human heart

    Energy Technology Data Exchange (ETDEWEB)

    Ekman, R.; Bjartell, A.; Lisander, J.; Edvinsson, L. (Univ. of Lund (Sweden))


    In patients undergoing aorto-coronary by-pass surgery, we found a 26% arterial-venous difference of immunoreactive {gamma}{sub 2}-melanocytostimulating hormone (MSH), a proopiomelanocortin (POMC) derived peptide known to possess profound hemodynamic effects. These results prompted an investigation of the presence of {gamma}{sub 2}-MSH in the human heart. Using a two-step extraction procedure, regions of human hearts were examined by sensitive and specific radioimmunoassays to determine their {gamma}{sub 2}-MSH content. Mean ({plus minus} SEM) concentrations of 0.14 {plus minus} 0.023 pmol/g and 0.12 {plus minus} 0.017 were found in right atrium and right ventricle, respectively. High performance liquid chromatography indicated that 80-90 % of the total immunoreactivity eluted in a single sharp peak in a position identical to that of synthetic {gamma}{sub 2}-MSH.

  6. ATF-2 immunoreactivity in post-mitotic and terminally differentiated human odontoblasts. (United States)

    Keklikoglu, Nurullah; Akinci, Sevtap


    Activating transcription factor 2 (ATF-2/CRE-BP1; cAMP-responsive element binding protein 1) is a member of nuclear transcription factor activator protein-1 (AP-1) family. AP-1 regulates cellular processes including growth, proliferation, differentiation and apoptosis. However, biological relationship of cellular process to each member of the AP-1 family is not clear yet. The objective of the present study was to compare the ATF-2 immunoreactivity in the post-mitotic and terminally differentiated odontoblasts and in the pulpal fibroblasts which can be divided by mitosis when required. Fibroblasts at various stages of differentiation co-exist in the human dental pulp. ATF-2 was investigated immunohistochemically in 20 permanent human teeth. According to the findings obtained, the mean percentage of ATF-2 positive cells was 68.5 ± 19.2% in the odontoblasts and 22.8 ± 13.7% in the pulpal fibroblasts. The comparison of ATF-2 positivity revealed a statistically significant difference between odontoblasts and pulpal fibroblasts. These findings have suggested that ATF-2 is more associated with cell survival rather than cell proliferation, and revealed much of effectiveness in maintaining terminal differentiation than the various differentiation stages of the cells. PMID:25417007

  7. Abdominal surgery activates nesfatin-1 immunoreactive brain nuclei in rats. (United States)

    Stengel, Andreas; Goebel, Miriam; Wang, Lixin; Taché, Yvette


    Abdominal surgery-induced postoperative gastric ileus is well established to induce Fos expression in specific brain nuclei in rats within 2-h after surgery. However, the phenotype of activated neurons has not been thoroughly characterized. Nesfatin-1 was recently discovered in the rat hypothalamus as a new anorexigenic peptide that also inhibits gastric emptying and is widely distributed in rat brain autonomic nuclei suggesting an involvement in stress responses. Therefore, we investigated whether abdominal surgery activates nesfatin-1-immunoreactive (ir) neurons in the rat brain. Two hours after abdominal surgery with cecal palpation under short isoflurane anesthesia or anesthesia alone, rats were transcardially perfused and brains processed for double immunohistochemical labeling of Fos and nesfatin-1. Abdominal surgery, compared to anesthesia alone, induced Fos expression in neurons of the supraoptic nucleus (SON), paraventricular nucleus (PVN), locus coeruleus (LC), Edinger-Westphal nucleus (EW), rostral raphe pallidus (rRPa), nucleus of the solitary tract (NTS) and ventrolateral medulla (VLM). Double Fos/nesfatin-1 labeling showed that of the activated cells, 99% were nesfatin-1-immunoreactive in the SON, 91% in the LC, 82% in the rRPa, 74% in the EW and VLM, 71% in the anterior parvicellular PVN, 47% in the lateral magnocellular PVN, 41% in the medial magnocellular PVN, 14% in the NTS and 9% in the medial parvicellular PVN. These data established nesfatin-1 immunoreactive neurons in specific nuclei of the hypothalamus and brainstem as part of the neuronal response to abdominal surgery and suggest a possible implication of nesfatin-1 in the alterations of food intake and gastric transit associated with such a stressor. PMID:19944727

  8. Comparative analysis of kisspeptin-immunoreactivity reveals genuine differences in the hypothalamic Kiss1 systems between rats and mice

    DEFF Research Database (Denmark)

    Overgaard, Agnete; Tena-Sempere, Manuel; Franceschini, Isabelle;


    kisspeptin-immunoreactivity in both nuclei and both sexes of rats and mice and quantified kisspeptin-immunoreactive nerve fibers. We also determined Kiss1 mRNA levels and measured kisspeptin-immunoreactivity in colchicine pretreated rats. Overall, we find higher levels of kisspeptin-immunoreactivity in the...

  9. Somatostatin-14-like immunoreactive neurons and fibres in the human olfactory bulb. (United States)

    Ohm, T G; Braak, E; Probst, A


    This study describes the morphological features and the distribution pattern of neurons in the human olfactory bulb which are immunoreactive for an antiserum against the neuropeptide somatostatin-14. Immunoreactive nerve cell bodies were mainly found in the white matter surrounding the cell clusters of the anterior olfactory nucleus. Some immunoreactive neurons were also found scattered throughout the anterior olfactory nucleus and the deeper parts of the inner granule cell layer. Only a few immunoreactive neurons were localized in the glomerular layer and the outer granule cell layer. Immunoreactive fibres were found in all layers of the olfactory bulb. In addition, an impressive number of coiled and kinked immunoreactive fibres were localized within the anterior olfactory nucleus forming a dense plexus. Accumulations of twisted and coiled branches of immunoreactive fibres were rarely found either surrounding or within the olfactory glomerula. The characteristics of somatostatin-14 immunoreactive neurons as seen in the combined pigment-Nissl preparation were studied after decolourizing the chromogen and restaining the preparations with aldehydefuchsin in order to demonstrate the lipofuscin pigment and gallocyanin chrome alum for Nissl material. About 90% of the immunoreactive neurons studied in this manner turned out to be devoid of lipofuscin granules. The remaining 10% displayed different patterns of pigmentation. These findings suggest the presence of different types of somatostatin-14-like immunoreactive neurons in the olfactory bulb of the human adult. PMID:2906788

  10. Enzyme immunoassay of immunoreactive trypsin in serum and blood spots

    International Nuclear Information System (INIS)

    An enzyme immunoassay method for the assay of serum immunoreactive trypsin (IRT) is described. The method is a two site binding assay carried out on microtitre plates as the solid phase. Wells were coated with affinity purified anti-human trypsin and bioinylated anti-trypsin and avidin-β-galactosidase were used as the second antibody and detection system respectively. The assay was sensitive enough to determine IRT concentrations in either serum or dried blood spots. A good correlation was obtained when the method was compared with the Hoechst radioimmunoassay method. (Author)

  11. The value of immunoreactive lipase in acute pancreatitis.



    We have evaluated a new agglutination test for serum immunoreactive lipase in 24 patients with abdominal pain and hyperamylasaemia. On admission all 20 patients with acute pancreatitis had a positive lipase test, 3 of the 4 patients who did not have pancreatitis had a negative lipase test. The sensitivity of the lipase test on day 1 is 100%, the specificity 96% and predictive value of a positive test is 95.2% compared to 83% for amylase. A negative test excludes pancreatitis. In addition, the...

  12. Immunoreactivity, sensory and physicochemical properties of fermented soy protein isolate. (United States)

    Meinlschmidt, Pia; Ueberham, Elke; Lehmann, Jörg; Schweiggert-Weisz, Ute; Eisner, P


    The effect of induced liquid state fermentation (Bacillus subtilis, Rhizopus oryzae, Saccharomyces cerevisiae, Lactobacillus helveticus) on the immunoreactivity, physicochemical and sensory properties of soy protein isolate (SPI) was studied. L. helveticus revealed the most abundant reduction in terms of immunoreactivity within soluble protein fractions, up to 100%, which could be measured by in vitro sandwich ELISA using mouse monoclonal anti-Glym5 antibodies (mAbs). Almost no binding was found in western blot analysis using mouse monoclonal mAbs and sera from soy sensitive individuals. Fermentation increased water- and oil-binding capacity as well as protein solubility at pH 4.0. Foaming activity was nearly doubled compared to non-fermented SPI. A decreased emulsifying capacity, foaming density, and quantity of soluble proteins at pH 7.0 were observed. Principal component analysis (PCA) confirmed decreased bitter and beany off-flavors of fermented samples compared to non-fermented SPI. Consequently, fermentation might be a promising method to produce tasty low-allergen food ingredients with good physicochemical properties. PMID:27006235

  13. Menin immunoreactivity in secretory granules of human pancreatic islet cells. (United States)

    Debelenko, Larisa V; Agarwal, Sunita; Du, Qiang; Yan, Wusheng; Erickson, Heidi S; Abu-Asab, Mones; Raffeld, Mark A; Libutti, Steven K; Marx, Stephen J; Emmert-Buck, Michael R


    The protein product of the Multiple Endocrine Neoplasia Type I (MEN1) gene is thought to be involved in predominantly nuclear functions; however, immunohistochemical (IHC) analysis data on cellular localization are conflicting. To further investigate menin expression, we analyzed human pancreas (an MEN1 target organ) using IHC analyses and 6 antibodies raised against full-length menin or its peptides. In 10 normal pancreas specimens, 2 independently raised antibodies showed unexpected cytoplasmic immunoreactivity in peripheral cells in each islet examined (over 100 total across all 10 patients). The staining exhibited a distinct punctate pattern and subsequent immunoelectron microscopy indicated the target antigen was in secretory granules. Exocrine pancreas and pancreatic stroma were not immunoreactive. In MEN1 patients, unaffected islets stained similar to those in normal samples but with a more peripheral location of positive cells, whereas hyperplastic islets and tumorlets showed increased and diffuse cytoplasmic staining, respectively. Endocrine tumors from MEN1 patients were negative for menin, consistent with a 2-hit loss of a tumor suppressor gene. Secretory granule localization of menin in a subset of islet cells suggests a function of the protein unique to a target organ of familial endocrine neoplasia, although the IHC data must be interpreted with some caution because of the possibility of antibody cross-reaction. The identity, cellular trafficking, and role of this putative secretory granule-form of menin warrant additional investigation. PMID:25153502

  14. Expression and immunoreactivity of HCV/HBV epitopes

    Institute of Scientific and Technical Information of China (English)

    Xin-Yu Xiong; Xiao Liu; Yuan-Ding Chen


    AIM: To develop the epitope-based vaccines to prevent Hepatitis C virus (HCV)/Hepatitis B virus (HBV) infections.METHODS: The HCV core epitopes C1 STNPKPQRKTKRNTNRRPQD (residuals aa2-21) and C2 VKFPGGGQIVGGVYLLPRR (residuals aa22-40), envelope epitope E GHRMAWDMMMNWSP (residuals aa315-328) and HBsAg epitope S CTTPAQGNSMFPSCCCTKPTDGNC (residuals aa124-147) were displayed in five different sites of the flock house virus capsid protein as a vector, and expressed in E. coli cells (pET-3 system).Immunoreactivity of the epitopes with anti-HCV and anti-HBV antibodies in the serum from hepatitis C and hepatitis B patients were determined.RESULTS: The expressed chimeric protein carrying the HCV epitopes C1, C2, E (two times), L3C1-I2E-L1C2-L2E could react with anti-HCV antibodies. The expressed chimeric protein carrying the HBV epitopes S, I3S could react with anti-HBs antibodies. The expressed chimeric proteins carrying the HCV epitopes C1, C2, E plus HBV epitope S, L3C1-I2E-L1C2-L2E-I3S could react with antiHCV and anti-HBs antibodies.CONCLUSION: These epitopes have highly specific and sensitive immunoreaction and are useful in the development of epitope-based vaccines.

  15. Radioimmunological determination and characterization of cathodal trypsin-like immunoreactivity in normal human plasma

    International Nuclear Information System (INIS)

    A radioimmunological method for determination of human cathodal trypsin-like immunoreactivity is described. DFP-treated human cathodal trypsin is used as standard and tracer. Freshly drawn normal human plasma contains about 25μg/l of cathodal trypsin-like immunoreactivity measured as DFP-treated cathodal trypsin. The normally circulating cathodal trypsin-like immunoreactivity is shown to consist mainly of cathodal trypsinogen. (Auth.)

  16. Hypothalamic CRF immunoreactivity in genetically hypothyroid (hyt/hyt) mice. (United States)

    Meserve, L A


    The induction of hypothyroidism in young rats by feeding thiouracil to their mothers during pregnancy has been shown to depress hypothalamic content of bioactive and immunoactive corticotropin-releasing factor (CRF). The present study was done to determine whether genetically hypothyroid young mice (hyt/hyt) born to euthyroid mothers (+/hyt) exhibited a similar depression in hypothalamic CRF immunoreactivity. Young euthyroid and hypothyroid littermate mice were examined by radioimmunoassay for hypothalamic CRF content at 15, 20, 25, or 30 days of age. Mean CRF content was depressed insignificantly (to about 80% of normal) by hypothyroidism, at 15-25 days of age. However, after weaning by the mother, 30-day-old hypothyroid pups demonstrated significantly depressed hypothalamic CRF levels (71%). It is suggested that maternal factors may be assisting in the maintenance of hypothalamic CRF until after weaning. Furthermore, genetic hypothyroidism does not appear to have nearly as marked an influence as thiouracil feeding on hypothalamic CRF levels. PMID:3496606

  17. Digoxin-like immunoreactivity in human body fluids

    International Nuclear Information System (INIS)

    The clinical and chemical characteristics of a solid-phase radioimmunoassay (RIA) for routine digoxin determination has been studied with the aim to confirm our previous observation of the presence of digoxin-like immunoreactive substance (DLIS) in serum (plasma) and urine of normal subjects not under digoxin treatment. The sensitivity of the assay was 2.1±0.6 pg/tube and the reproducibility, tested with two different urine pools in terms of digoxin-equivalents (d.e.), was 12.5% (285.6±35.7 pg/ml d.e., n=19) and 20.6% (123.8±25.5 pg/ml d.e., n=19), respectively. The mean DLIS concentration in the blood of 32 normal subjects was 15.6±8.0 pg/ml d.e. (range 0-60 pg/ml d.e.). The mean DLIS concentration in urine of 37 normal subjects (overnight collection) was 160.0±52.3 pg/ml d.e. (range 70-350 pg/ml d.e.), while the mean 24-hour DLIS excretion of 10 normal subjects was 97.3±39.7 ng d.e. Two urine pools were extracted with organic solvents. Good recoveries (80-100%) were obtained with methanol, while poor recoveries were obtained with methylene chloride, hexane and petroleum ether. The present study indicates that DLIS is not a large charged molecule, neither salt, nor fatty acid, which are considered the most frequent non-specific interferences in RIA systems. Urine samples may be more useful for pathophysiological studies on digoxin-like immunoreactivity in human body fluids, because of their higher DLIS concentrations (4-10 times the concetration in blood)

  18. Tyrosine hydroxylase immunoreactivity is common in the enteric nervous system in teleosts. (United States)

    Olsson, Catharina


    Tyrosine hydroxylase (TH) is the rate-limiting enzyme in the synthesis of catecholamines and TH immunoreactivity is indicative of cells synthesising either adrenaline/noradrenaline or dopamine. In this study, the distribution of TH immunoreactivity was examined in two distantly related teleost species, zebrafish (Danio rerio) and shorthorn sculpin (Myoxocephalus scorpius). In both species, TH-immunoreactive nerve cell bodies and varicose nerve fibres were common in the myenteric plexus of the intestine. However, no TH-immunoreactive nerve cell bodies were seen in the sculpin stomach. The TH-immunoreactive nerve cell bodies seemed to constitute a larger proportion of the total enteric population in shorthorn sculpin (50 ± 5 %, n = 3067 cells) compared with zebrafish (14 ± 2 %, n = 10,163 cells). In contrast, in sculpin, the TH-immunoreactive cells were smaller than the average enteric nerve cell bodies, whereas in zebrafish, the relationship was the opposite. In developing zebrafish larvae, TH-immunoreactive nerve cell bodies were common (approx. 75 % of the total population) at 3 days post-fertilization (dpf), but decreased in numbers between 3 and 7 dpf. In conclusion, in contrast to previous studies, TH-immunoreactive intrinsic neurons are common in the fish gut. Their role and function need to be further characterized in order to understand the potential importance of this enteric subpopulation in controlling various gut functions. PMID:26572541

  19. CD34 immunoreactivity and interstitial cells of Cajal in the human and mouse gastrointestinal tract

    DEFF Research Database (Denmark)

    Vanderwinden, J M; Rumessen, J J; De Laet, M H;


    Immunoreactivity for the tyrosine kinase receptor Kit (Kit-ir) is an established marker for the interstitial cells of Cajal (ICC) of the gut. Recently, the presence of CD34 immunoreactivity (CD34-ir) has been reported in Kit-ir ICC around the myenteric plexus in human small intestine. Conversely...

  20. Serum immunoreactive erythropoietin in HIV-infected patients

    International Nuclear Information System (INIS)

    Serum immunoreactive erythropoietin (SIE) and hemoglobin levels were measured in 152 patients infected with the human immunodeficiency virus. Anemia was present in 18% of asymptomatic patients who tested positive for the human immunodeficiency virus, 50% of patients with a condition related to the acquired immunodeficiency syndrome (AIDS), and 75% of patients with AIDS. The mean SIE level for untreated AIDS patients was greater than for patients who tested positive for human immunodeficiency virus or patients with an AIDS-related condition but not outside the normal range for SIE, and the incremental increase in SIE level for a given decline in hemoglobin level was much less in AIDS patients than in patients with uncomplicated iron deficiency anemia. Forty-two patients were treated with zidovudine, and the hemoglobin level fell 10 g/L or more in 48%. The data indicate that SIE level is inappropriately low in anemic AIDS patients. The ability of these patients to produce erythropoietin is intact and can be expressed with zidovudine therapy. However, even very high levels of SIE fail to stimulate erythropoiesis adequately

  1. Androgen receptor immunoreactivity in rat occipital cortex after callosotomy

    Directory of Open Access Journals (Sweden)

    G Lepore


    Full Text Available Gonadal steroidogenesis can be influenced by direct neural links between the central nervous system and the gonads. It is known that androgen receptor (AR is expressed in many areas of the rat brain involved in neuroendocrine control of reproduction, such as the cerebral cortex. It has been recently shown that the occipital cortex exerts an inhibitory effect on testicular stereoidogenesis by a pituitary-independent neural mechanism. Moreover, the complete transection of the corpus callosum leads to an increase in testosterone (T secretion of hemigonadectomized rats. The present study was undertaken to analyze the possible corticocortical influences regulating male reproductive activities. Adult male Wistar rats were divided into 4 groups: 1 intact animals as control; 2 rats undergoing sham callosotomy; 3 posterior callosotomy; 4 gonadectomy and posterior callosotomy. Western blot analysis showed no remarkable variations in cortical AR expression in any of the groups except in group I where a significant decrease in AR levels was found. Similarly, both immunocytochemical study and cell count estimation showed a lower AR immunoreactivity in occipital cortex of callosotomized rats than in other groups. In addition, there was no difference in serum T and LH concentration between sham-callosotomized and callosotomized rats. In conclusion, our results show that posterior callosotomy led to a reduction in AR in the right occipital cortex suggesting a putative inhibiting effect of the contralateral cortical area.

  2. Serum immunoreactive erythropoietin in HIV-infected patients

    Energy Technology Data Exchange (ETDEWEB)

    Spivak, J.L.; Barnes, D.C.; Fuchs, E.; Quinn, T.C. (Johns Hopkins Univ. School of Medicine, Baltimore, MD (USA))


    Serum immunoreactive erythropoietin (SIE) and hemoglobin levels were measured in 152 patients infected with the human immunodeficiency virus. Anemia was present in 18% of asymptomatic patients who tested positive for the human immunodeficiency virus, 50% of patients with a condition related to the acquired immunodeficiency syndrome (AIDS), and 75% of patients with AIDS. The mean SIE level for untreated AIDS patients was greater than for patients who tested positive for human immunodeficiency virus or patients with an AIDS-related condition but not outside the normal range for SIE, and the incremental increase in SIE level for a given decline in hemoglobin level was much less in AIDS patients than in patients with uncomplicated iron deficiency anemia. Forty-two patients were treated with zidovudine, and the hemoglobin level fell 10 g/L or more in 48%. The data indicate that SIE level is inappropriately low in anemic AIDS patients. The ability of these patients to produce erythropoietin is intact and can be expressed with zidovudine therapy. However, even very high levels of SIE fail to stimulate erythropoiesis adequately.

  3. Comparison of immunoreactive serum trypsinogen and lipase in Cystic Fibrosis

    International Nuclear Information System (INIS)

    The incidence of Cystic Fibrosis (CF) is 1 in 2,000. Early detection and treatment of CF may necessitate newborn screening with a reliable and cost-effective test. Serum immunoreactive trypsinogen (IRT) an enzyme produced by the pancreas, is detectable by radioimmunoassay (RIA) techniques. Recently, it has been shown that IRT is elevated in CF infants for the first few months of life and levels become subnormal as pancreatic insufficiency progresses. Other enzymes produced by the pancreas, such as lipase, are also elevated during this time. The author's earlier work confirmed previous reports of elevated IRT levels in CF infants. The development of a new RIA for lipase (nuclipase) has enabled comparison of these 2 pancreatic enzymes in C.F. Serum IRT and lipase determinations were performed on 2 groups of CF patients; infants under 1 year of age, and children between 1 and 18 years of age. Control populations of the same age groups were included. The results showed that both trypsin (161 +- 92 ng/ml, range 20 to 400) and lipase (167 +- 151 ng/ml, range 29 to 500) are elevated in CF in the majority of infants. Control infants had values of IRT ranging from 20 to 29.5 ng/ml and lipase values ranging from 23 to 34 ng/ml. IRT becomes subnormal in most CF patients by 8 years of age as pancreatic function insufficiency increases. Lipase levels and IRT levels correlate well in infancy, but IRT is a more sensitive indicator of pancreatic insufficiency in older patients with CF

  4. Distribution of hypocretin (orexin) immunoreactivity in the feline pons and medulla. (United States)

    Zhang, Jian Hua; Sampogna, Sharon; Morales, Francisco R; Chase, Michael H


    The distribution of hypocretin-1 (hcrt-1) and hypocretin-2 (hcrt-2) immunoreactivities in the cat brainstem was examined using immunohistochemical techniques. Hcrt-1- and hcrt-2-positive fibers with varicosities were detected in almost all brainstem regions. However, no hcrt-1- or hcrt-2-immunoreactive neuronal somata were observed in the cat brainstem. Both hcrt-1- and hcrt-2-labeled fibers exhibited different densities in distinct regions of the brainstem. In most brainstem regions, the intensity of hcrt-1 immunoreactivity was higher than that of hcrt-2 immunoreactivity. The highest densities of hcrt-1- and hcrt-2-positive fibers were found in the nucleus raphe dorsalis (RD), the laterodorsal tegmental nucleus (LDT) and the locus coeruleus (LC), suggesting an important role for these peptides in functions related to sleep-wake behavior. PMID:14672810

  5. GnRH-immunoreactive centrifugal visual fibers in the Nile crocodile (Crocodylus niloticus). (United States)

    Médina, Monique; Repérant, Jacques; Miceli, Dom; Ward, Roger; Arckens, Lutgarde


    Thin varicose centrifugal visual fibers, between 30-45 in number and displaying cGnRH-I immunoreactivity, were identified in Crocodylus niloticus. Approximately 80% of these fibers were also FMRF-amide-like immunoreactive. The cGnRH-I fibers extended from the preoptic region to the retina where they appeared to terminate in the external portion of the inner plexiform layer. The location of their neurons of origin could not be determined precisely following the intraocular injection of the retrograde axonal tracer RITC. Nevertheless, the presence of cGnRH-I-immunoreactive neurons exclusively within the complex comprising the terminal nerve and the septo-preoptic region, and of several retinopetal fibers labelled retrogradely with the axonal tracer at the septo-preoptic junction, indicates that the cGnRH-immunoreactive centrifugal visual system originates from within this complex. PMID:16002052

  6. FMRFamide immunoreactivity in the nervous system of the medusa Polyorchis penicillatus

    DEFF Research Database (Denmark)

    Grimmelikhuijzen, C J; Spencer, A N


    with several antisera to oxytocin/vasopressin and bombesin/gastrin-releasing peptide. The morphology and location of most FMRFamide-immunoreactive neurons in Polyorchis coincides with two identified neuronal systems, which have been recently discovered from neurophysiological studies....

  7. Humoral immunoreactivity to gliadin and to tissue transglutaminase is present in some patients with multiple myeloma

    Directory of Open Access Journals (Sweden)

    Matkovic Suzana


    Full Text Available Abstract Background Multiple myeloma (MM is a clonal B-cell disorder with many immunological disturbances. The aim of this work was to assess whether some of food antigens contribute to the imbalance of immune response by screening the sera of MM patients for their immunoreactivity to food constituent gliadin, to tissue transglutaminase-2 (tTG-2 and to Ro/SSA antigen. Sera from 61 patients with MM in various stages of disease, before, or after some cycles of conventional therapy were analyzed by commercial Binding Site ELISA tests. The control group consisted of 50 healthy volunteers. Statistical analysis of data obtained was performed by Mann Whitney Test. Results The higher serum IgA immunoreactivity to gliadin was found in 14/56 patients and in one of control people. The enhanced serum IgG immunoreactivity to gliadin was found in only two of tested patients and in two controls. The enhanced IgA immunoreactivity to tTG-2 was found in 10/49 patients' sera, while 4/45 patients had higher serum IgG immunoreactivity. The enhanced serum IgG immunoreactivity to RoSSÀ antigen was found in 9/47 analyzed MM patients' sera. Statistical analysis of data obtained revealed that only the levels of anti-tTG-2 IgA immunoreactivity in patients with MM were significantly higher than these obtained in healthy controls (P Conclusion Data obtained showed the existence of the enhanced serum immunoreactivity to gliadin, tTG-2 and Ro/SSA antigens in some patients with MM. These at least partially could contribute to the immunological imbalance frequently found in this disease.

  8. Somatostatin-like immunoreactivity in non-pyramidal neurons of the human isocortex. (United States)

    Braak, E; Braak, H; Weindl, A


    The distribution of somatostatin-immunoreactive cell bodies and axons throughout the human isocortex and subjacent white matter was examined. Vibratome sections of cortical tissue (30-40 micrometers thick) obtained at surgery were treated to reveal the antigen by the unlabelled antibody enzyme method. Two types of somatostatin-immunoreactive axons were present: short, coiled axons and extended ones that follow a straight course in various directions. Somatostatin immunoreactive nerve cell bodies were encountered in layers II-VI and in the subjacent white matter. The majority of labelled cells were found in the white matter and layer VI, and then in layers II and III. The immunoreactive perikarya were fusiform, triangular or multipolar in shape and did not show preferential orientation of their long axis. Frequently, the fusiform neurons in layer VI and in the white matter were aligned parallel to radiate bundles of myelinated fibres. The immunoreactive neurons gave rise to a few thick dendrites. Often thin axon-like processes could also be recognized, originating either from the cell body or from a thicker dendrite. After destaining of the chromogen and counterstaining with aldehydefuchsin and gallocyanin chromealum, the formerly immunoreactive neurons displayed a light and eccentrically located nucleus. The soma contained only a sparse amount of basophilic substance and was nearly devoid of lipofuscin granules. In electron micrographs, the cisterns of the rough endoplasmic reticulum (RER) were localized near the periphery of the soma. Immunoreactivity occurred along membranes of the RER cistern, outer mitochondrial membrane, and in particles 120-150 micrometers in diameter. Rounded areas (up to a diameter of 1 micrometer) lacked immunoreactivity. Furthermore, there were a few tiny lysosomes. PMID:2867717

  9. Diurnal variation of β-endorphin like immunoreactivity in rat brain, pituitary gland, and plasma

    International Nuclear Information System (INIS)

    β-endorphin like immunoreactivity was measured in the brain, pituitary gland and plasma of rats at 2 A.M, 8 A.M, 2 P.M and 8 P.M. Values were higher in the brain and pituitary gland at 8 P.M and in the plasma at 8 A.M and 2 P.M. The findings suggest a circadian rhythm in the production and release of β-endorphin immunoreactive material. (Author)

  10. Brain interleukin 1 and S-100 immunoreactivity are elevated in Down syndrome and Alzheimer disease.


    Griffin, W S; Stanley, L C; Ling, C; White, L.; MacLeod, V; Perrot, L J; White, C.L.; Araoz, C


    Interleukin 1, an immune response-generated cytokine that stimulates astrocyte proliferation and reactivity (astrogliosis), was present in up to 30 times as many glial cells in tissue sections of brain from patients with Down syndrome and Alzheimer disease compared with age-matched control subjects. Most interleukin 1-immunoreactive glia in Down syndrome and Alzheimer disease were classified as microglia. The number of interleukin 1 immunoreactive neurons did not appear to differ in Down synd...

  11. Neuropeptide Y-like immunoreactive neurons in the human olfactory bulb. (United States)

    Ohm, T G; Braak, E; Probst, A; Weindl, A


    Neuropeptide Y-like (NPY) immunoreactivity was localized in the adult human olfactory bulb by the unlabeled antibody enzyme (peroxidase anti-peroxidase; PAP) technique in vibratome sections. The majority of NPY-immunoreactive somata was localized in the white matter surrounding the anterior olfactory nucleus. Immunoreactive neurons were less numerous within the anterior olfactory nucleus and within the olfactory bulb layers. NPY-immunoreactive fibres were present in the white matter, the anterior olfactory nucleus, and in the olfactory bulb layers. Fibres within the white matter were generally aligned in a straight path parallel to the long axis of the olfactory bulb and tract. Fibres within the anterior olfactory nucleus showed no clear orientation and displayed numerous branching points. Coiled plexus of NPY-immunoreactive fibres were present in the glomerular layer of the olfactory bulb. Additional characteristics of the NPY-immunoreactive neurons were studied after decolouring the chromogen and restaining the sections with aldehydefuchsin to demonstrate the presence of lipofuscin granules and also with gallocyanin chrome alum to stain the Nissl substance. This analysis showed that the neurons belong to the class of non-pigmented nerve cells. PMID:3251589

  12. Study on development and localization of CTGF-immunoreactive cells in central nervous system of rats

    Institute of Scientific and Technical Information of China (English)

    SU Bing-yin; CAI Wen-qin; ZHANNG Cheng-gang; B.Perbal


    Objective: To study the development of connective tissue growth factor(CTGF) immunoreactive cells in the central nervous system (CNS) of E8-P300 rats. Methods: Immunocytochemistry was employed in our study. Results: No CTGF-immunoreactive cells were detected in the CNS of rats during prenatal stages. A few of CTGF-positive cells were detected in the early postnatal stage. However, the positive cells increased gradually in later stages. CTGF-immunoreactive cells widely distributed in the CNS of rats in the first 30 to 60 days postnatally, and the density of immunoreactive products was the highest in these days. The number and staining intensity of CTGF-positive cells decreased and their area of distribution diminished gradually with age. The positive cells included neurons mainly located in the cingulate cortex,striatum, hippocampus, hypothalamus and cerebellum, and astrocytes in white matter of the spinal cord and ependymal cells of the brain. Most of CTGF-immunoreactive cells were quite big in size with a long process. Conclusion: CTGF-immunoreactive cells were found in the CNS of rats, and their numbers and positive signal decreased with the age.

  13. Localization of atrial natriuretic peptide mRNA and immunoreactivity in the rat heart and human atrial appendage

    Energy Technology Data Exchange (ETDEWEB)

    Hamid, Q.; Wharton, J.; Terenghi, G.; Hassall, C.J.S.; Aimi, J.; Taylor, K.M.; Nakazato, H.; Dixon, J.E.; Burnstock, G.; Polak, J.M.


    The localization of mRNA encoding preproatrial natriuretic peptide was investigated in tissue sections and cultures of rat heart and in sections of human right atrial appendage using the technique of in situ hybridization with /sup 32/P- and /sup 35/S-labeled RNA probes. Rat atrial natriuretic peptide (ANP) transcripts were demonstrated in numerous atrial myocytes and, to a lesser extent, in ventricular myocytes in both tissue sections and newborn rat heart cultures. These findings are consistent with those obtained by RNA blot analysis of rat heart total RNA, indicating that a single prepro-ANP transcript of approx. 900 nucleotides was present in the ventricles as well as the atria. Using a /sup 35/S-labeled RNA probe for human ANP mRNA, ANP transcripts were also localized to the majority of myocytes in the human right atrial appendage. Only background levels of autoradiographic labeling were obtained when RNA probes identical to the coding sequence of rat or human ANP mRNA were used. A close correlation was found between the distribution of ANP immunoreactivity and prepro-ANP mRNA in these preparations. These results provide unequivocal evidence for the expression of the ANP gene in the rat ventricles, as well as the atria, because myocytes in these tissues have been established as the sites of both ANP localization and precursor biosynthesis. The combined use of cardiac cultures and in situ hybridization may be of value in future studies investigating the regulation of ANP synthesis in cardiac myocytes.

  14. Pretreatment P53 immunoreactivity does not infer radioresistance in prostate cancer patients

    International Nuclear Information System (INIS)

    Purpose: To test, in a clinical context, the hypothesis that p53 aberrations, assessed by immunoreactivity, are related to radioresistance as suggested by several experimental studies. Methods and Materials: Sixty patients with prostate cancer who underwent transurethral resection of the prostate or biopsy prior to definitive external beam therapy were retrospectively identified. The endpoint in the study was cancer specific survival. The nuclear accumulation of the aberrant p53 protein was evaluated by immunohistochemistry with the pantropic, monoclonal Ab-6 anti-p53 antibody (clone DO-1) on pretreatment biopsies. Immunoreactivity was related to stage, grade, and cancer-specific survival. Results: There was a correlation between p53 immunoreactivity and low tumor stage (p < 0.001), but no relation between p53 status and grade was found. Moreover, no significant difference was found in cancer-specific survival between the p53 positive tumors (109 months) and the p53 negative tumors (99 months). Conclusions: No disadvantage regarding survival was seen for patients with p53 immunoreactive tumors, implicating that p53 immunoreactivity does not infer radioresistance in prostate cancer. This suggests that the p53 inactivation may be a less important determinant of tumor response to radiotherapy in some human cancers than in the previously studied experimental situations. Thus, other mechanisms may be more important in determining outcome after radiation. However, the series is small and data should be interpreted with caution

  15. Effects of heat and high-pressure treatments on the solubility and immunoreactivity of almond proteins. (United States)

    Zhang, Yan; Zhang, Jieqiong; Sheng, Wei; Wang, Shuo; Fu, Tong-Jen


    The effects of dry and moist heat, autoclave sterilization and high-pressure treatment on the biochemical characteristics and immunological properties of almond proteins were investigated. Changes in the solubility and immunoreactivity of almond proteins extracted from treated almond flour were evaluated using a total protein assay, indirect competitive inhibition enzyme-linked immunosorbent assay (IC-ELISA), and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Almond proteins were stable during dry-heat treatment at temperatures below 250°C. Dry heat at 400°C, boiling, autoclave sterilization and high-pressure treatment in the presence of water at ⩾ 500 MPa greatly reduced the solubility and immunoreactivity of almond proteins. SDS-PAGE revealed that the protein profiles of almond flour samples treated under these conditions also changed significantly. The synergistic effects of heat, pressure and the presence of water contributed to significant changes in solubility and immunoreactivity of almond proteins. PMID:26776044

  16. Immunoreactivity for Choline Acetyltransferase of Peripheral-Type (pChAT) in the Trigeminal Ganglion Neurons of the Non-Human Primate Macaca fascicularis

    International Nuclear Information System (INIS)

    Transcripts of the choline acetyltransferase (ChAT) gene reveal a number of different splice variants including ChAT of a peripheral type (pChAT). Immunohistochemical staining of the brain using an antibody against pChAT clearly revealed peripheral cholinergic neurons, but failed to detect cholinergic neurons in the central nervous system. In rodents, pChAT-immunoreactivity has been detected in cholinergic parasympathetic postganglionic and enteric ganglion neurons. In addition, pChAT has been observed in non-cholinergic neurons such as peripheral sensory neurons in the trigeminal and dorsal root ganglia. The common type of ChAT (cChAT) has been investigated in many parts of the brain and the spinal cord of non-human primates, but little information is available about the localization of pChAT in primate species. Here, we report the detection of pChAT immunoreactivity in trigeminal ganglion (TG) neurons and its co-localization with Substance P (SP) and/or calcitonin gene-related peptide (CGRP) in the cynomolgus monkey, Macaca fascicularis. Neurons positive for pChAT were observed in a rather uniform pattern in approximately half of the trigeminal neurons throughout the TG. Most pChAT-positive neurons had small or medium-sized cell bodies. Double-immunofluorescence staining showed that 85.1% of SP-positive cells and 74.0% of CGRP-positive cells exhibited pChAT immunoreactivity. Most pChAT-positive cells were part of a larger population of neurons that co-expressed SP and/or CGRP

  17. Morphometric characteristics of Neuropeptide Y immunoreactive neurons of human cortical amygdaloid nucleus

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    Mališ Miloš


    Full Text Available Introduction Cortical amygdaloid nucleus belongs to the corticomedial part of the amygdaloid complex. In this nucleus there are neurons that produce neuropetide Y. This peptide has important roles in sleeping, learning, memory, gastrointestinal regulation, anxiety, epilepsy, alcoholism and depression. Material and methods We investigated morphometric characteristics (numbers of primary dendrites, longer and shorter diameters of cell bodies and maximal radius of dendritic arborization of NPY immunoreactive neurons of human cortical amygdaloid nucleus on 6 male adult human brains, aged 46 to 77 years, by immunohistochemical avidin-biotin technique. Results Our investigation has shown that in this nucleus there is a moderate number of NPY immunoreactive neurons. 67% of found neurons were nonpyramidal, while 33% were pyramidal. Among the nonpyramidal neurons the dominant groups were multipolar neurons (41% - of which 25% were multipolar irregular, and 16% multipolar oval. Among the pyramidal neurons the dominant groups were the neurons with triangular shape of cell body (21%. All found NPY immunoreactive neurons (pyramidal and nonpyramidal altogether had intervals of values of numbers of primary dendrites 2 to 6, longer diameters of cell bodies 13 to 38 µm, shorter diameters of cell bodies 9 to 20 µm and maximal radius of dendritic arborization 50 to 340 µm. More than a half of investigated neurons (57% had 3 primary dendrites. Discussion and conclusion The other researchers did not find such percentage of pyramidal immunoreactive neurons in this amygdaloid nucleus. If we compare our results with the results of the ather researchers we can conclude that all pyramidal NPY immunoreactive neurons found in this human amygdaloid nucleus belong to the class I of neurons, and that all nonpyramidal NPY immunoreactive neurons belong to the class II of neurons described by other researchers. We suppose that all found pyramidal neurons were projectional.

  18. Increased density of DISC1-immunoreactive oligodendroglial cells in fronto-parietal white matter of patients with paranoid schizophrenia. (United States)

    Bernstein, Hans-Gert; Jauch, Esther; Dobrowolny, Henrik; Mawrin, Christian; Steiner, Johann; Bogerts, Bernhard


    Profound white matter abnormalities have repeatedly been described in schizophrenia, which involve the altered expression of numerous oligodendrocyte-associated genes. Transcripts of the disrupted-in-schizophrenia 1 (DISC1) gene, a key susceptibility factor in schizophrenia, have recently been shown to be expressed by oligodendroglial cells and to negatively regulate oligodendrocyte differentiation and maturation. To learn more about the putative role(s) of oligodendroglia-associated DISC1 in schizophrenia, we analyzed the density of DISC1-immunoreactive oligodendrocytes in the fronto-parietal white matter in postmortem brains of patients with schizophrenia. Compared with controls (N = 12) and cases with undifferentiated/residual schizophrenia (N = 6), there was a significantly increased density of DISC1-expressing glial cells in paranoid schizophrenia (N = 12), which unlikely resulted from neuroleptic treatment. Pathophysiologically, over-expression of DISC1 protein(s) in white matter oligodendrocytes might add to the reduced levels of two myelin markers, 2',3'-cyclic-nucleotide 3'-phosphodiesterase and myelin basic protein in schizophrenia. Moreover, it might significantly contribute to cell cycle abnormalities as well as to deficits in oligodendroglial cell differentiation and maturation found in schizophrenia. PMID:26315603

  19. A case of bilateral renal cell carcinoma associated with long-term dialysis showing false-positive immunoreactivity for TFE3 as Xp11 translocation renal cell carcinoma. (United States)

    Kurisaki-Arakawa, Aiko; Saito, Tsuyoshi; Takahashi, Michiko; Mitani, Keiko; Fukumura, Yuki; Nagashima, Yoji; Argani, Pedrum; Yao, Takashi


    Renal carcinomas associated with Xp11.2 translocations/transcription factor 3 (TFE3) gene fusion (Xp11 translocation RCC) are a rare subtype of renal cell carcinoma. A middle-aged Japanese man, who had a medical history of dialysis for more than 12 years, had bilateral renal cancers with a background of acquired cystic disease of the kidney and remarkable deposition of calcium oxalate in the tumorous area. The right renal tumor showed papillary architecture of clear cells with diffuse and strong immunoreactivity for TFE3 and focal and weak positivity for cathepsin K, suggesting a possibility of Xp11 translocation RCC. However, RT-PCR failed to detect any type of the reported fusion genes involving TFE3. Thus, the sample was sent for a TFE3 break-apart FISH assay in a renal tumor consultation service, which reported no evidence of TFE3 gene rearrangement. The right renal tumor was finally diagnosed as papillary renal cell carcinoma with cystic change. We report here a case of bilateral renal cell carcinoma in a patient undergoing long-term dialysis, which showed false-positive immunoreactivity for TFE3 immunostaining. Titration of TFE3 immunohistochemical staining (IHC) should be performed and cross-referenced with the FISH or RT-PCR results to avoid the misinterpretation of TFE3 IHC results. PMID:24228124

  20. Gamma-melanocyte-stimulating hormone-like immunoreactivity in blood cells of human eosinophilic patients. (United States)

    Johansson, O; Virtanen, M; Hilliges, M; Hansson, L O


    The immunohistochemical localization of the peptide gamma-melanocyte-stimulating hormone (gamma-MSH) within human polymorphonuclear leucocytes of blood from eosinophilic patients is described. The gamma-MSH immunoreactivity was observed only in neutrophilic granulocytes leaving all other cell types immuno-negative. PMID:1805488

  1. FMRFamide immunoreactivity is generally occurring in the nervous systems of coelenterates

    DEFF Research Database (Denmark)

    Grimmelikhuijzen, C J


    Abundant FMRFamide immunoreactivity has been found in the nervous systems of all hydrozoan, anthozoan, scyphozoan and ctenophoran species that were looked upon. This general and abundant occurrence shows that FMRFamide-like material must play a crucial role in the functioning of primitive nervous...

  2. Gastrin/CCK-like immunoreactivity in the nervous system of coelenterates

    DEFF Research Database (Denmark)

    Grimmelikhuijzen, C J; Sundler, F; Rehfeld, J F


    Using immunocytochemistry, gastrin/CCK-like immunoreactivity is found in sensory nerve cells in the ectoderm of the mouth region of hydra and in nerve cells in the endoderm of all body regions of the sea anemone tealia. These results are corroborated by radioimmunoassay: One hydra contains at least...

  3. Associative learning down-regulates PKCβ2- and γ-immunoreactivity in astrocytes

    NARCIS (Netherlands)

    Zee, E.A. van der; Kronforst-Collins, M.A.; Disterhoft, J.F.


    We showed previously that associative learning induced a twofold increase in protein kinase Cγ-immunoreactivity (PKCγ-ir) in rabbit CA1 pyramidal neurons, whereas subicular neurons remained unchanged. Here, we investigated the effects of associative learning on PKC-positive astrocytes by determining

  4. FMRF-amide-like immunoreactivity in brain and pituitary of the hagfish Eptatretus burgeri (Cyclostomata)

    DEFF Research Database (Denmark)

    Jirikowski, G; Erhart, G; Grimmelikhuijzen, C J;


    the hypothalamus to the olfactory system and caudally to the medulla oblongata. FMRF-amide-like immunoreactivity was also found in cells of the adenohypophysis. These observations suggest that the hagfish possesses a brain FMRF-amide-like transmitter system and pituitary cells containing FMRF-amide-like material...

  5. Effects of intraduodenal administration of HCl and glucose on circulating immunoreactive secretin and insulin concentrations. (United States)

    Boden, G; Essa, N; Owen, O E; Reichle, F A


    A new radioimmunoassay for secretin was used to investigate (a) serum secretin responses to intraduodenally infused HCl and glucose, (b) the metabolic half-life and the volume of distribution of exogenous secretin and (c) the effect of endogenously released secretin on insulin secretion in 25 anesthetized dogs. Portal and femoral venous blood samples were taken simultaneously before, during, and after intraduodenal infusion of HCl (21 meq/30 min) and glucose (131 ml/30 min). Control experiments were performed with intraduodenal infusion of saline. Mean portal venous immunoreactive secretin concentration of six dogs rose from 313 muU/ml before to 1,060 muU/ml 10 min after initiation of the intestinal acidification (P dogs mean portal venous immunoreactive insulin concentration rose from 38 muU/ml before to 62 muU/ml at the end of the infusion (P Pancreatic exocrine function was studied in four dogs. The rise in secretin concentration was followed promptly by a highly significant increase in exocrine pancreatic flow rate and bicarbonate secretion, indicating biological activity of the circulating immunoreactive secretin. The effect of intraduodenal infusion of glucose on immunoreactive secretin concentration was studied in 12 dogs. Glucose in concentrations ranging from 2.5% to 10% had no detectable influence on portal or peripheral secretin concentration. Infusion of 50% glucose caused a slight decline in secretin concentration. The metabolic clearance rate, half-life of disappearance, and volume of distribution of exogenous secretin was studied in three dogs by the constant infusion technic. The metabolic clearance rate was 730+/-34 ml/min, volume of distribution was 17.4+/-0.8% of body weight, and the half-life of disappearance was 2.8+/-0.1 min. It could be calculated that 1.38 U/kg-h(-1) of endogenous secretin was released into the peripheral circulation during the steady state period of the HCl infusion experiments. The data indicated that immunoreactive

  6. Direct reticular projections of trigeminal sensory fibers immunoreactive to CGRP: potential monosynaptic somatoautonomic projections

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    W Michael ePanneton


    Full Text Available Few trigeminal sensory fibers project centrally beyond the trigeminal sensory complex, with only projections of fibers carried in its sensory anterior ethmoidal (AEN and intraoral nerves described. Fibers of the AEN project into the brainstem reticular formation where immunoreactivity against substance P and CGRP are found. We investigated whether the source of these peptides could be from trigeminal ganglion neurons by performing unilateral rhizotomies of the trigeminal root and looking for absence of label. After an 8-14d survival, substance P immunoreactivity in the trigeminal sensory complex was diminished, but we could not conclude that the sole source of this peptide in the lateral parabrachial area and lateral reticular formation arises from primary afferent fibers. Immunoreactivity to CGRP after rhizotomy however was greatly diminished in the trigeminal sensory complex, confirming the observations of others. Moreover, CGRP immunoreactivity was nearly eliminated in fibers in the lateral parabrachial area, the caudal ventrolateral medulla, both the peri-ambiguus and ventral parts of the rostral ventrolateral medulla, in the external formation of the nucleus ambiguus, and diminished in the caudal pressor area. The nearly complete elimination of CGRP in the lateral reticular formation after rhizotomy suggests this peptide is carried in primary afferent fibers. Moreover, the arborization of CGRP immunoreactive fibers in these areas mimics that of direct projections from the AEN. Since electrical stimulation of the AEN induces cardiorespiratory adjustments including an apnea, peripheral vasoconstriction, and bradycardia similar to those seen in the mammalian diving response, we suggest these perturbations of autonomic behavior are enhanced by direct somatic primary afferent projections to these reticular neurons. We believe this to be first description of potential direct somatoautonomic projections to brainstem neurons regulating autonomic

  7. Oxaliplatin-induced loss of phosphorylated heavy neurofilament subunit neuronal immunoreactivity in rat DRG tissue

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    Connor Bronwen


    Full Text Available Abstract Background Oxaliplatin and related chemotherapeutic drugs cause painful chronic peripheral neuropathies in cancer patients. We investigated changes in neuronal size profiles and neurofilament immunoreactivity in L5 dorsal root ganglion (DRG tissue of adult female Wistar rats after multiple-dose treatment with oxaliplatin, cisplatin, carboplatin or paclitaxel. Results After treatment with oxaliplatin, phosphorylated neurofilament heavy subunit (pNF-H immunoreactivity was reduced in neuronal cell bodies, but unchanged in nerve fibres, of the L5 DRG. Morphometric analysis confirmed significant changes in the number (-75%; P P P = 0.82, NF-M (-1%, P = 0.96 or NF-H (0%; P = 0.93 after oxaliplatin treatment, although the sizes of parvalbumin (-29%, P = 0.047, NF-M (-11%, P = 0.038 and NF-H (-28%; P = 0.0033 immunoreactive neurons were reduced. In an independent comparison of different chemotherapeutic agents, the number of pNF-H-immunoreactive neurons was significantly altered by oxaliplatin (-77.2%; P P = 0.03 but not by carboplatin or paclitaxel, and their mean cell body area was significantly changed by oxaliplatin (-31.1%; P = 0.008 but not by cisplatin, carboplatin or paclitaxel. Conclusion This study has demonstrated a specific pattern of loss of pNF-H immunoreactivity in rat DRG tissue that corresponds with the relative neurotoxicity of oxaliplatin, cisplatin and carboplatin. Loss of pNF-H may be mechanistically linked to oxaliplatin-induced neuronal atrophy, and serves as a readily measureable endpoint of its neurotoxicity in the rat model.

  8. An investigation of appetite-related peptide transcript expression in Atlantic cod (Gadus morhua) brain following a Camelina sativa meal-supplemented feeding trial. (United States)

    Tuziak, Sarah M; Rise, Matthew L; Volkoff, Hélène


    Camelina sativa is a hardy oilseed crop with seeds that contain high levels of ω3 polyunsaturated fatty acids and protein, which are critical components of fish feed. Camelina might thus be used as a cheaper and more sustainable supplement to fish-based products in aquaculture. Atlantic cod, Gadus morhua, is a species of interest in the aquaculture industry due to a decrease in wild populations and subsequent collapse of some cod fisheries. As cod are carnivorous fish, it is necessary to determine how this species physiologically tolerates plant-based diets. In this study, juvenile Atlantic cod were subjected to 13 weeks of either 15 or 30% camelina meal (CM)-supplemented diets or a control fish meal feed. Growth and food intake were evaluated and the mRNA expression of appetite-related hormones [pro-melanin-concentrating hormone (pmch), hypocretin (synonym: orexin, hcrt), neuropeptide Y (npy) and cocaine- and amphetamine-regulated transcript (cart)] was assessed using quantitative real-time PCR in brain regions related to food intake regulation (telencephalon/preoptic area, optic tectum/thalamus and hypothalamus). CM inclusion diets caused decreases in both growth and food intake in Atlantic cod. Optic tectum pmch transcript expression was significantly higher in fish fed the 30% CM diet compared to fish fed the 15% CM diet. In the hypothalamus, compared to fish fed the control diet, hcrt expression was significantly higher in fish fed the 30% CM diet, while npy transcript expression was significantly higher in fish fed the 15% CM diet. cart mRNA expression was not affected by diet in any brain region. Further studies are needed to determine which factors (e.g. anti-nutritional factors, palatability and nutritional deficits) contribute to reduced feed intake and growth, as well as the maximum CM inclusion level that does not negatively influence feed intake, growth rate and the transcript expression of appetite-related factors in Atlantic cod. PMID:25151310

  9. Distribution and Origin of VIP-, SP-, and Phospholipase Cβ2 -Immunoreactive Nerves in the Tongue of the Bullfrog, Rana catesbeiana. (United States)

    Tadokoro, Osamu; Ando, Hiroshi; Kawahara, Ichiro; Asanuma, Naokazu; Okumura, Masayo; Kitagawa, Junichi; Kondo, Eiji; Yagasaki, Hiroshi


    Previous studies have found a few intralingual ganglionic cells that were immunoreactive to vasoactive intestinal polypeptide (VIP) in the frog. A recent study reported a large number of such cells, and the possibility of the release of substance P (SP) from these. The aim of the present study was to investigate the distribution, origin, and colocalization of VIP- and SP- immunoreactive nerves in the tongue of the bullfrog, R. catesbeiana. In addition, the study also examined the colocalization of SP and phospholipase Cβ2 (PLCβ2 ) in the tongue and jugular ganglion. VIP immunoreactivity was seen in unipolar cells that were sparse in nerve bundles in the submucosal and muscle layers. The density of VIP-immunoreactive cells was approximately 4.8 cells/mm(3) . Their fibers terminated in the vicinity of the epithelial basal layer of the fungiform papillae. SP immunoreactivity was not seen in the VIP-immunoreactive cells, but was observed in pseudounipolar cells in the jugular ganglion. The SP fibers terminated close to the free surface, showing spindle- and button-like profiles. Transection of glossopharyngeal nerve resulted in the persistence of VIP-immunoreactive cells and the disappearance of SP-immunoreactive fibers in the tongue. SP immunoreactivity was co-expressed with PLCβ2 in both the tongue and jugular ganglia. No PLCβ2 immunoreactivity was seen in cells comprising the epithelial taste disk. These findings indicate that the origin of VIP nerve fibers are unipolar cells in the tongue, and SP and PLCβ2 fibers originate from pseudounipolar cells that may be able to release SP primarily in the jugular ganglion. Anat Rec, 299:929-942, 2016. © 2016 Wiley Periodicals, Inc. PMID:26916909

  10. Responses of plasma cyclic AMP, serum immunoreactive insulin, C-peptide immunoreactivity and blood sugar levels to glucagon in patients with liver diseases.

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    Full Text Available Levels of plasma cyclic AMP, serum immunoreactive insulin (IRI, serum c-peptide immunoreactivity (CPR and blood sugar (BS were determined 0, 15, 30, 45 and 60 min after a glucagon injection (0.01 mg per kg body weight in normal controls, patients with acute hepatitis and liver cirrhosis. Plasma cyclic AMP responses to glucagon in liver disease patients varied widely in peak value, and only in patients with fulminant hepatitis and decompensated liver cirrhosis with poor prognosis was the response suppressed. The peak response of BS was found significantly later in liver cirrhosis patients than in normal controls. IRI and CPR responses to glucagon were lower in acute hepatitis patients than in normal controls and liver cirrhosis patients. IRI levels and their sum were also lower in acute hepatitis patients, although CPR levels were not significantly different. Thus, the ratio of the sum of CPR from 0 to 60 min to that of IRI was significantly higher in acute hepatitis, indicating impaired pancreatic secretion of insulin to glucagon stimulation as well as increased uptake of insulin by the liver in acute hepatitis.

  11. A longitudinal study of maternal digoxin-like immunoreactive substances in normotensive pregnancy and pregnancy-induced hypertension. (United States)

    Kerkez, S A; Poston, L; Wolfe, C D; Quartero, H W; Carabelli, P; Petruckevitch, A; Hilton, P J


    The serum of women in the third trimester of pregnancy demonstrates cross-reactivity with some commercially available antibodies to digoxin. A number of studies have suggested that levels of this digoxin-like immunoreactive substance(s) are further increased in patients with pregnancy-induced hypertension, and some have proposed that the digoxin-like immunoreactive substance could be useful as a predictor of pregnancy-induced hypertension. We measured digoxin-like immunoreactive substance levels every 2 weeks throughout the third trimester in 170 women; of these, 20 developed hypertension. Digoxin-like immunoreactive substance levels rose with gestational age. A graph of the slope of digoxin-like immunoreactive substance plotted against gestational age was fitted for the results obtained from each woman. There was no significant difference in the mean rate of increase of digoxin-like immunoreactive substance level per week between pregnancy-induced hypertension and normotensive pregnancy, nor was there any difference between these two groups at any gestational age studied. These results suggest that measuring digoxin-like immunoreactive substance levels is not useful as a predictor of pregnancy-induced hypertension. PMID:2316589

  12. Somatostatin-like immunoreactivity is found in dendritic guard cells of human sweat ducts. (United States)

    Johansson, O; Hilliges, M; Wang, L


    Somatostatin is reported in a new population of human sweat duct cells. The epithelial content of somatostatin-like immunoreactivity in normal human skin from various sites of the body was investigated using indirect immunofluorescence. With this methodology we found round-to-oval somatostatin-immunoreactive cells situated outside the lining sweat duct cells at the level of the stratum spinosum. The cells had processes that were clearly directed towards the duct lumen, passing between the lining duct cells. This finding raises new questions and ideas about somatostatin's role in the skin, and could point to a possible involvement of somatostatin in immune defense, sweat secretion modification, antiproliferation, or other actions. PMID:8097870

  13. Presumptive FMRF-amide-like immunoreactive retinopetal fibres in Crocodylus niloticus. (United States)

    Médina, Monique; Repérant, Jacques; Ward, Roger; Miceli, Dom


    A small contingent of 30-50 of centrifugal visual fibres, showing FMRF-amide-like immunoreactivity, has been identified in C. niloticus; these fibres extend from the chiasmatic region into the retina. They do not take the marginal optic tract, but pass medially to the chiasmatic fascicles, from the preoptic region. The cells of origin of these fibres have not been identified. However, none of the retinopetal neurons of the brainstem [M. Medina, J. Reperant, R. Ward, D. Miceli, Centrifugal visual system of Crocodylus niloticus : a hodological, histochemical and immunocytochemical study, J. Comp. Neurol. 468 (2004) 65-85], labelled by retrograde transport of rhodamine beta-isothiocyanate after intraocular injection of this tracer, show FMRF-amide-like immunoreactivity; neither are any of the FMRF-amide-like immunopositive neurons in the crocodile brain, particularly those of the complex involving the terminal nerve and the septo-preoptic region, labelled by rhodamine after its intraocular injection. PMID:15464765

  14. Sodium channel Nav1.8 immunoreactivity in painful human dental pulp

    Directory of Open Access Journals (Sweden)

    Tate S


    Full Text Available Abstract Background The tetrodotoxin-resistant voltage-gated sodium channel Nav1.8 (SNS1/PN3 is expressed by nociceptors and may play a role in pain states. Methods Using specific antibodies for immunohistochemistry, we studied Nav1.8 – immunoreactivity in human dental pulp in relation to the neuronal marker neurofilament. Human tooth pulp was extracted from teeth harvested from a total of twenty-two patients (fourteen without dental pain, eight patients with dental pain. Results Fibres immunoreactive for Nav1.8, were significantly increased on image analysis in the painful group: median (range Nav1.8 to Neurofilament % area ratio, non-painful 0.059 (0.006–0.24, painful 0.265 (0.13–0.5, P = 0.0019. Conclusion Nav1.8 sodium channels may thus represent a therapeutic target in trigeminal nerve pain states.

  15. Different pattern of haemagglutinin immunoreactivity of equine influenza virus strains isolated in Poland

    Directory of Open Access Journals (Sweden)

    Kwaśnik Małgorzata


    Full Text Available The immunoreactivity of haemagglutinin (HA polypeptides of equine influenza virus was compared among the strains isolated in Poland, using H3 monoclonal antibody. A stronger signal in immunoblot reaction was observed for A/equi/Pulawy/2008 HA polypeptides compared to A/equi/Pulawy/2006, despite the fact that both strains are phylogenetically closely related and belong to Florida clade 2 of American lineage. The strongest signal, observed in the case of A/equi/Pulawy/2008, seemed to be connected with the presence of G135, I213, E379, and/or V530 instead of R135, M213, G379, and I530 present in A/equi/Pulawy/2006 HA sequence. This implies that point mutations within amino acid sequences of HA polypeptides of equine influenza virus may change their immunoreactivity even when they are not located within five basic antigenic sites.

  16. Ablation of prion protein immunoreactivity by heating in saturated calcium hydroxide

    Directory of Open Access Journals (Sweden)

    Holtzapple Mark T


    Full Text Available Abstract Background Prions, the infectious agents that cause transmissible spongiform encephalopathies (TSEs, are relatively resistant to destruction by physical, enzymatic, and chemical treatments. Hydrolysis in boiling saturated calcium hydroxide (limewater utilizes inexpensive chemicals to digest protein components of offal. The purpose of this work was to determine if incubating brain material from scrapie-infected sheep in near-boiling saturated calcium hydroxide solution (Ca(OH2 would abolish immunoreactivity of the infectious prion (PrPSc as determined by western blot. Findings After incubating for as few as 10 minutes in saturated calcium hydroxide at 99°C, immunoreactivity of protease resistant bands by western blot analysis is completely lost. Conclusion Boiling in limewater may offer an alternative for disposal of carcasses and enable alternative uses for rendered products from potentially infected carcasses.

  17. Orexin (hypocretin)-like immunoreactivity in the cat hypothalamus: a light and electron microscopic study. (United States)

    Zhang, J H; Sampogna, S; Morales, F R; Chase, M H


    Orexin-A-like immunoreactive (OrA-ir) neurons and terminals in the cat hypothalamus were examined using immunohistochemical techniques. OrA-ir neurons were found principally in the lateral hypothalamic area (LHA) at the level of the tuberal cinereum and in the dorsal and posterior hypothalamic areas. In the LHA the majority of the neurons were located dorsal and lateral to the fornix; a small number of OrA-ir neurons were also present in other regions of the hypothalamus. OrA-ir fibers with varicose terminals were detected in almost all hypothalamic regions. The high density of fibers was located in the suprachiasmatic nucleus, the infundibular nucleus (INF), the tuberomamillary nucleus (TM) and the supra- and pre-mamillary nuclei. Ultrastructural analysis revealed that OrA-ir neurons in the LHA receive abundant input from non-immunoreactive terminals. These terminals, which contained many small, clear, round vesicles with a few large, dense core vesicles, made asymmetrical synaptic contacts with OrA-ir dendrites, indicating that the activity of orexin neurons is under excitatory control. On the other hand, the terminals of OrA-ir neurons also made asymmetrical synaptic contact with dendrites in the LHA, the INF and the TM. The dendrites in the LHA were both non-immunoreactive and OrA-ir; conversely, the dendrites in the INF and the TM were non-immunoreactive. In these regions, OrA-ir terminals contained many small, clear, round vesicles with few large, dense core vesicles, suggesting that orexinergic neurons also provide excitatory input to other neurons in these regions. PMID:11204055

  18. GABA immunoreactive elements in the sensory system of the earthworm Eisenia fetida (Annelida, Clitellata)


    G Kiszler; E Várhalmi; L Krecsák; Z Solt; E Pollák; Molnár, L.


    The presence of γ-aminobutyric acid (GABA) immunoreactive elements was observed in the sensory system of Eisenia fetida (Annelida, Clitellata). Among the primary sensory cells a high number of labelled cells was found in the epithelium. Using whole-mount preparations and multispectral recording, the number and the distribution pattern of the immunopositive cells were determined in different body segments. Our morphological analysis revealed four typical types of stained primary ...

  19. p53 immunoreaction in endoscopic biopsy specimens of colorectal cancer, and its prognostic significance.


    A. Yamaguchi; Nakagawara, G.; Kurosaka, Y.; Nishimura, G.; Yonemura, Y.; Miyazaki, I.


    The expression of p53 protein was immunohistochemically studied in formalin-fixed paraffin-embedded biopsy specimens of 203 colorectal carcinomas by use of a monoclonal antibody specific for the p53 protein. PAb1801. p53 protein expression with its reactivity localised in nuclei was found in 121 (59.6%) of the cancers. There was no correlation of p53 immunoreactivity with histological classification, wall invasion, lymphatic invasion, venous invasion, lymph node metastases, or peritoneal meta...

  20. Gene expression analysis in the parvalbumin-immunoreactive PV1 nucleus of the mouse lateral hypothalamus


    Girard, F; Mészár Zoltán (1977-) (állatorvos); Marti, C.; Davis, F. P.; Celio, M.


    A solitary, elongated cluster of parvalbumin-immunoreactive neurons has been previously observed in the rodent ventrolateral hypothalamus. However, the function of this so-called PV1 nucleus is unknown. In this article, we report the results of an unbiased, broad and in-depth molecular characterization of this small, compact group of neurons. The Allen Brain Atlas database of in situ hybridization was screened in order to identify genes expressed in the PV1-nucleus-containing area of the hypo...

  1. 413 Effect of Hydrolysis and Polymerization on Bovine Beta-lactoglobulin Immunoreactivity


    Villas-Boas, Mariana Battaglin; Sabadin, Isabele Serimarco; de Lima Zollner, Ricardo; Netto, Flavia Maria


    Background Enzymatic treatments such as hydrolysis with proteases and polymerization using transglutaminase (TG) have been studied to reduce the immunoreactivity of β-Lactoglobulin (β-Lg). Bromelain is a cisteine protease that is not usually used for production of hypoallergenic hydrolysates. TG is an enzyme that catalyses the formation of inter and intramolecular isopeptide bonds between glutamine and lysine residues. The present study is aimed at investigating the antigenic response of β-Lg...

  2. Developmental and Regional Patterns of GAP-43 Immunoreactivity in a Metamorphosing Brain


    Simmons, Andrea Megela; Tanyu, Leslie H.; Horowitz, Seth S.; Chapman, Judith A.; Brown, Rebecca A.


    Growth-associated protein-43 is typically expressed at high levels in the nervous system during development. In adult animals, its expression is lower, but still observable in brain areas showing structural or functional plasticity. We examined patterns of GAP-43 immunoreactivity in the brain of the bullfrog, an animal whose nervous system undergoes considerable reorganization across metamorphic development and retains a strong capacity for plasticity in adulthood. Immunolabeling was mostly d...

  3. Dopamine- and Tyrosine Hydroxylase-Immunoreactive Neurons in the Brain of the American Cockroach, Periplaneta americana (United States)

    Hamanaka, Yoshitaka; Minoura, Run; Nishino, Hiroshi; Miura, Toru; Mizunami, Makoto


    The catecholamine dopamine plays several vital roles in the central nervous system of many species, but its neural mechanisms remain elusive. Detailed neuroanatomical characterization of dopamine neurons is a prerequisite for elucidating dopamine’s actions in the brain. In the present study, we investigated the distribution of dopaminergic neurons in the brain of the American cockroach, Periplaneta americana, using two antisera: 1) an antiserum against dopamine, and 2) an antiserum against tyrosine hydroxylase (TH, an enzyme required for dopamine synthesis), and identified about 250 putatively dopaminergic neurons. The patterns of dopamine- and TH-immunoreactive neurons were strikingly similar, suggesting that both antisera recognize the same sets of “dopaminergic” neurons. The dopamine and TH antibodies intensively or moderately immunolabeled prominent brain neuropils, e.g. the mushroom body (memory center), antennal lobe (first-order olfactory center) and central complex (motor coordination center). All subdivisions of the mushroom body exhibit both dopamine and TH immunoreactivity. Comparison of immunolabeled neurons with those filled by dye injection revealed that a group of immunolabeled neurons with cell bodies near the calyx projects into a distal region of the vertical lobe, which is a plausible site for olfactory memory formation in insects. In the antennal lobe, ordinary glomeruli as well as macroglomeruli exhibit both dopamine and TH immunoreactivity. It is noteworthy that the dopamine antiserum labeled tiny granular structures inside the glomeruli whereas the TH antiserum labeled processes in the marginal regions of the glomeruli, suggesting a different origin. In the central complex, all subdivisions excluding part of the noduli and protocerebral bridge exhibit both dopamine and TH immunoreactivity. These anatomical findings will accelerate our understanding of dopaminergic systems, specifically in neural circuits underlying aversive memory

  4. Effects of mutations on enzyme activity and immunoreactivity of the S1 subunit of pertussis toxin.


    Lobet, Y; Cieplak, W; Smith, S. G.; Keith, J M


    By introducing a series of six different substitutions at and around position 9, we investigated the structural requirements of the amino-terminal region of the S1 subunit of pertussis toxin for both enzyme activity and immunoreactivity. All mutant S1 analogs with a substitution at this location exhibited severely decreased ADP-ribosyltransferase activity (range, 400- to 2,500-fold). In contrast, alteration of arginine 58 had considerably less effect. The reactivity of the mutant molecules wi...

  5. Protein gene product 9.5-immunoreactive nerve fibres and cells in human skin. (United States)

    Wang, L; Hilliges, M; Jernberg, T; Wiegleb-Edström, D; Johansson, O


    Sections of human skin were processed according to the indirect immunofluorescence technique with a rabbit antiserum against human protein gene product 9.5 (PGP 9.5). Immunoreactivity was detected in intraepidermal and dermal nerve fibres and cells. The intraepidermal nerves were varicose or smooth with different diameters, running as single processes or branched, straight or bent, projecting in various directions and terminating in the stratum basale, spinosum or granulosum. The density of the intraepidermal nerves varied between the different skin areas investigated. PGP 9.5-containing axons of the lower dermis were found in large bundles. They separated into smaller axon bundles within the upper dermis, entering this portion of the skin perpendicular to the surface. Then they branched into fibres mainly arranged parallel to the epidermal-dermal junctional zone. However, the fibres en route to the epidermis traversed the upper dermis more or less perpendicularly. Furthermore, immunoreactive dermal nerve fibres were found in the Meissner corpuscles, the arrector pili muscles, hair follicles, around the eccrine and apocrine sweat glands and around certain blood vessels. Such fibres were also observed around most subcutaneous blood vessels, sometimes heavily innervating these structures. Numerous weakly-to-strongly PGP 9.5-immunoreactive cells were found both in the epidermis and in the dermis. PMID:2143435

  6. Simultaneous detection of Clavibacter michiganensis subsp. nebraskensis and Pantoea stewartii subsp. stewartii based on microsphere immunoreaction. (United States)

    Zhang, Fan; Li, Jinfeng; Zou, Mingqiang; Chen, Yan; Wang, Yanfei; Qi, Xiaohua


    Clavibacter michiganensis subsp. nebraskensis (Cmn) and Pantoea stewartii subsp. stewartii (Pss) are two plant pathogens that can cause tremendous agricultural economic losses. This novel method based on microsphere immunoreaction was developed for the simultaneous detection of Cmn and Pss in maize. This multiplex method was constructed based on microsphere immunodetection with fluorescent labels such as quantum dots (QDs) and R-phycoerythrin (R-PE) for the detection of Cmn and Pss. Captured QDs and R-PE serve as signal reporters for fluorescent readout. The principle of this method is based on a sandwich immunoreaction. Cmn and Pss captured by the microspheres were detected using flow cytometry. The limit of detection of this method was 10 times lower than the enzyme-linked immunosorbent assay (ELISA), and its analysis time (1 h) was much shorter compared with ELISA (6-8 h). The method, which has been proven to be an effective approach to multiplex detection of plant bacteria (Cmn and Pss as models), not only increased the varieties but also improved the sensitivity. The microsphere immunoreaction provides a universal method for the multiplex determination of microbes because of its high sensitivity, specificity, and speed. In the future, the method will be more fully validated in vivo to detect diversiform bacteria. PMID:23169888

  7. Oenanthe javanica extract increases immunoreactivities of antioxidant enzymes in the rat kidney

    Institute of Scientific and Technical Information of China (English)

    Hyun-Jin Tae; Joon Ha Park; Jeong-Hwi Cho; In Hye Kim; Ji Hyeon Ahn; Jae Chul Lee; Jong-Dai Kim


    Background Oenanthe javanica is an aquatic perennial herb originated from East Asia.Nowadays,the effects of Oenanthe javanica have been proven in various disease models.Studies regarding the antioxidant effect of Oenanthe javanica in the kidney are still unclear.Methods This study was therefore performed to investigate the effect of the Oenanthe javanica extract (OJE) in the rat kidney using immunohistochemistry for antioxidant enzymes,copper,zinc-superoxide dismutase (SOD1),manganese superoxide dismutase (SOD2),catalase (CAT) and glutathione peroxidase (GPx).Sprague-Dawley rats were randomly assigned to three groups:(1) normal diet fed-group (normal-group),(2) diet containing ascorbic acid (AA)-fed group (AA-group) as a positive control,(3) diet containing OJE-fed group (OJE-group).AA and OJE were supplied during 28 days.Results The side-effects were not observed in all the groups.Immunoreactivities of SOD1,SOD2,CAT and GPx were easily detected in the distal tubules of the kidney,and their immunoreactivities in the AA-and OJE-groups were increased to about 1.4-1.5 times and 2 times,respectively,compared with those in the normal-group.Conclusion OJE significantly increased expressions of SOD1 & 2,CAT and GPx immunoreactivities in the distal tubules of the rat kidney,and this finding suggests that significant enhancements of endogenous enzymatic antioxidants by OJE treatment may be a legitimate strategy for decreasing oxidative stresses in the kidney.

  8. Heterogeneity of human plasma insulin: techniques for separating immunoreactive components and their determination by radioimmunoassay

    International Nuclear Information System (INIS)

    When human plasma is filtered on Sephadex G-SO fine, insulin immunoreactivity is recovered in two peaks: 'big insulin', the higher molecular weight component and 'little insulin', the lower molecular component, having elution volumes that correspond to those of porcine proinsulin 125I and porcine insulin 125I respectively. The presence of another form of immunoreactive insulin 'big big insulin' was detected from an insuloma suspect and its elution pattern corresponding to serum albumin. The eluates correspondent to 'big' and 'little' insulin as well as 'big big' component were assayed by radioimmunoassay using crystalline human insulin as a standard, porcine insulin 125 tracer and anti insulin serum. The antibody, raised in guinea-pigs, was sensitive and potent being adequate for the assay. The reactivity of insulin and proinsulin was tested against the antibody. The relative proportions of several components of total immunoreactive insulin in plasma were studied in basal conditions in five normal subjects and in the patient JSC with pancreatic insulin-secreting tumor as well as after glucose stimuli in all tolbutamide in JSC. (author)

  9. Evaluation of residual immunoreactivity in red and white wines clarified with gluten or gluten derivatives. (United States)

    Cattaneo, A; Ballabio, C; Bertelli, A A E; Fiocchi, A; Galli, C L; Isoardi, P; Terracciano, L; Restani, P


    Gluten or hydrolyzed gluten could be a suitable alternative to animal proteins in the wine clarification process, but their residues could represent a risk for individuals suffering from coeliac disease or allergic to cereal proteins. The aim of this study was to investigate the presence of gluten in wines treated with gluten or its hydrolysate in the clarification process and to assess its antigenicity in commercial products. The presence of residual immunoreactive gluten was evaluated by electrophoresis (SDS-PAGE) and immunoblotting. Data obtained in several red and white wine samples showed that no residue was detectable in any of the red wines. In white wines, gluten reduced the protein content less completely, but most samples showed no immunoreactivity after the wine had been treated with gluten or its derivatives, either alone or combined with bentonite, silica gel or tannins. The use of gluten derivatives coupled with bentonite was the most effective method of removing immunoreactive protein in white wines. In conclusion, the use of gluten derivatives in wine clarification seems to exclude a risk for subjects susceptible to coeliac disease or gluten allergy. However, it is recommended that wine producers continuously monitor the clarification process in order to protect the most sensitive individuals. PMID:14518594

  10. Brain interleukin 1 and S-100 immunoreactivity are elevated in Down syndrome and Alzheimer disease. (United States)

    Griffin, W S; Stanley, L C; Ling, C; White, L; MacLeod, V; Perrot, L J; White, C L; Araoz, C


    Interleukin 1, an immune response-generated cytokine that stimulates astrocyte proliferation and reactivity (astrogliosis), was present in up to 30 times as many glial cells in tissue sections of brain from patients with Down syndrome and Alzheimer disease compared with age-matched control subjects. Most interleukin 1-immunoreactive glia in Down syndrome and Alzheimer disease were classified as microglia. The number of interleukin 1 immunoreactive neurons did not appear to differ in Down syndrome and Alzheimer disease compared with control brain. Numerous temporal lobe astrocytes in Alzheimer disease and postnatal Down syndrome were intensely interleukin 1-, S-100-, and glial fibrillary acidic protein-immunoreactive and had reactive structure. Interleukin 1 levels in Alzheimer disease temporal lobe homogenates were elevated, as were the levels of S-100 and glial fibrillary acidic protein, two proteins reportedly elevated in reactive astrocytes. These data suggest that increased expression of S-100 in Down syndrome, resulting from duplication of the gene on chromosome 21 that encodes the beta subunit of S-100, may be augmented by elevation of interleukin 1. As a corollary, the astrogliosis in Alzheimer disease may be promoted by elevation of interleukin 1. Images PMID:2529544

  11. Age-related alterations in immunoreactivity of the midsized neurofilament subunit in the brainstem reticular formation of the cat. (United States)

    Zhang, J H; Sampogna, S; Morales, F R; Chase, M H


    In the present study, we compared the immunoreactivity of the midsized subunit of neurofilaments (NF-M) in the brainstem reticular formation of adult and old cats. There was a dramatic decrease in immunoreactivity in most reticular nuclei in the old cats. The most obvious reduction in these regions occurred in dendritic arborizations. In contrast, a small number of nuclei showed a slight increase in immunoreactivity in the aged animals. The age-related changes in immunoreactivity indicate that there is an alteration of NF-M content in reticular neurons and their processes in old age. Such changes in NF-M content may be the basis for the alterations in the morphology of reticular neurons in aged animals. PMID:9374292

  12. The pattern of FMRFamide and serotonin immunoreactive elements in the nervous system of Aspidogaster conchicola K. Baer, 1827 (Aspidogastrea, Aspidogastridae)


    Tolstenkov, Oleg; Terenina, Nadezhda; Kreshchenko, Natalia; Gustafsson, Margaretha


    The patterns of the neuropeptide FMRFamide and serotonin (5-HT) immunoreactive (IR) elements in the nervous system of Aspidogaster conchicola (Aspidogastrea, Aspidogastridae) are described using immunocytochemistry and confocal scanning laser microscopy. Both FMRFamid and 5-HT immunoreactivities occur in the bilobed brain, the three pairs of longitudinal nerve cords and many transverse commissures. The adhesive disc is strongly innervated by FMRFamide-IR nerve fibres. Many 5-HT-IR neurones we...

  13. Neuropeptide Y-like immunoreactivity in rat cranial parasympathetic neurons: coexistence with vasoactive intestinal peptide and choline acetyltransferase

    International Nuclear Information System (INIS)

    Neuropeptide Y (NPY) is widely distributed in the sympathetic nervous system, where it is colocalized with norepinephrine. The authors report here that NPY-immunoreactive neurons are also abundant in three cranial parasympathetic ganglia, the otic, sphenopalatine, and ciliary, in the rat measured by radioimmunoassay. High-performance liquid chromatographic analysis of the immunoreactive material present in the otic ganglion indicates that this material is very similar to porcine NPY and indistinguishable from the NPY-like immunoreactivity present in rat sympathetic neurons. These findings raise the possibility that NPY acts as a neuromodulator in the parasympathetic as well as the sympathetic nervous system. In contrast to what had been observed for sympathetic neurons, NPY-immunoreactive neurons in cranial parasympathetic ganglia do not contain detectable catecholamines or tyrosine hydroxylase immunoreactivity, and many do contain immunoreactivity for vasoactive intestinal peptide and/or choline acetyltransferase. These findings suggest that there is no simple rule governing coexpression of NPY with norepinephrine, acetylcholine, or vasoactive intestinal peptide in autonomic neurons. Further, while functional studies have indicated that NPY exerts actions on the peripheral vasculature which are antagonistic to those of acetylcholine and vasoactive intestinal peptide, the present results raise the possibility that these three substances may have complementary effects on other target tissues

  14. Localization of neuropeptide-Y immunoreactivity in estradiol-concentrating cells in the hypothalamus

    International Nuclear Information System (INIS)

    Considerable evidence shows that gonadal steroids exert a facilitatory influence on levels and release of neuropeptide-Y (NPY) from the hypothalamus. However, it is not known whether gonadal steroids act directly on NPY-producing cells in the arcuate nucleus (ARC) of the hypothalamus to produce these facilitatory effects on NPY or whether they act on other cells that have a modulatory influence via synapses on ARC NPY cells. We applied the combined method of steroid autoradiography and immunocytochemistry to assess the localization of [3H]estradiol in relation to NPY-producing cells in the hypothalamus. Rats (n = 6) were bilaterally ovariectomized and injected intracerebroventricularly with colchicine. Twenty-four hours later each rat received an iv injection of 17 beta-[2,4,6,7,16,17(-3)H]estradiol (SA, 166 Ci/mmol) at a dose of 5.0 micrograms/kg BW. One hour after the injection of [3H]estradiol, the rats were perfused with 4% paraformaldehyde; brains were removed, frozen in isopentane precooled in liquid nitrogen (-190 C), sectioned, and processed for autoradiography. The autoradiograms were then incubated with specific antibodies for NPY immunostaining by the avidin-biotin-peroxidase method. The results revealed NPY-immunopositive cells in the ARC, striatum, hippocampus, amygdala, and cerebral cortex and a few cells in the median eminence. NPY-immunoreactive fibers were also detected in the internal layer of the median eminence. The largest number of neurons showing NPY immunoreactivity in the cytoplasm was detected in the ARC, and only in this nucleus did we observed colocalization of [3H]estradiol and NPY immunoreactivity in neurons. A population of NPY-immunopositive cells in the ARC (10-20%) exhibited nuclear [3H]estradiol; the majority of these cells were located in the lateral and ventral portions of the ARC

  15. Serum antibody immunoreactivity to equine zona protein after SpayVac vaccination. (United States)

    Mask, Tracy A; Schoenecker, Kathryn A; Kane, Albert J; Ransom, Jason I; Bruemmer, Jason E


    Immunocontraception with porcine ZP (pZP) can be an effective means of fertility control in feral horses. Previous studies suggest that antibodies produced after pZP vaccination may both inhibit fertilization and cause follicular dysgenesis. Zonastat-H, PZP-22, and SpayVac are three pZP vaccines proposed for use in horses. Although all these vaccines contain the pZP antigen, variations in antigen preparation and vaccine formulation lead to differences in antigenic properties among them. Likewise, despite numerous efficacy and safety studies of Zonastat-H and PZP-22, the contraceptive mechanisms of SpayVac remain unclear. The preparation of pZP for SpayVac is thought to include more nonzona proteins, making it less pure than the other two vaccines. This may result in increased antigenicity of the vaccine. We therefore investigated the immunoreactivity of serum antibodies from SpayVac-vaccinated mares to equine zona protein. Western blot analyses revealed an immunoreactivity of these antibodies to protein isolated from mature equine oocytes, ZP, follicular tissues, and ovarian tissues. Immunohistochemical analyses were used to locate the binding of serum antibodies to the ZP of immature oocytes in ovarian stromal tissue. We also found serum antibodies from SpayVac-treated mares to be predominantly specific for zona protein 3. Collectively, our results suggest a model where serum antibodies produced in response to SpayVac vaccination are immunoreactive to equine zona protein in vitro. Our study lends insight into the contraceptive mechanisms underlying the infertility observed after SpayVac vaccination. PMID:25922172

  16. Immunohistochemical distribution of Calbindin D-28K immunoreactivity in the central nervous system of adult cat

    Institute of Scientific and Technical Information of China (English)

    LIU Tao; LI Jin-lian; XIONG Kang-hui; LI Ji-shuo


    Objective: In order to get more information about the possible functions of Calbindin D-28K in the central nervous system of adult cat, the distribution of Calbindin D-28K in the central nervous system of adult cat was examined. Methods: Immunohistochemical staining techniques were used, and immunostained sections were observed under a light microscopy. Results: A high density of both immunoreactive perikarya and fibers were observed in the basal ganglia, amygdaloid complex, nucleus of the fields of Forel, subthalamic nucleus, paracentral nucleus, pulvinar nucleus, subthalamus, dorsal hypothalamic area, lateral hypothalamic area, anterior hypothalamus, suprachiasmatic nucleus, superior colliculus, inferior colliculus, oculomo-tor nucleus, superior olivary complex, marginal nucleus of the brachium conjunctivum, vestibular nuclei, the spinal trigeminal nucleus, nucleus of the solitary tract, cuneate nucleus, inferior olivary complex, dorsal motor nucleus of the vagus nerve, the molecular layer of the cerebellum, the purkinje cell layer of the cerebellum and in the laminae Ⅱ of the spinal cord, whereas the dentate gyrus, the central medial nucleus of the thalamus, the paracentral and central lateral nucleus of the thalamus, the lateral dorsal nucleus of the thalamus,the ventrolateral complex of the thalamus, the medioventral nucleus of the thalamus, the posterior hypothalamic area, the dorsal hypothalamic area, the infundibular nucleus, the dorsomedial hypothalamic nucleus and the interfascicular nucleus had just a high density of immunoreactive perikarya, and no positive fibres were detected in these areas. Conclusion: The present results showed that Calbindin D-28K-like immunoreactivity was widely distributed throughout the central nervous system of adult cat and might play an important role in the activities of the neurons in the central nervous system of adult cat.

  17. The Spectrum of Hormone Immunoreactivity in Typical and Atypical Pituitary Adenomas

    Directory of Open Access Journals (Sweden)

    Yeşim ERTAN


    Full Text Available Objective: We aimed to assess the spectrum of hormone immunoreactivity in our pituitary adenoma cases and discuss the diagnostic parameters of atypical pituitary adenomas.Material and Methods: A total of 166 pituitary adenoma cases diagnosed from 2005 to 2008 in our department were included in the present study. Hematoxylin-eosin stained and immunohistochemistry performed slides (ACTH, PRL, GH, TSH, FSH, LH, Ki-67, and p53 were evaluated. Cases having more than two mitoses on 10 high power fields besides more than 3% Ki-67 index were accepted in the atypical group.Results: Histologically, 159 cases were typical pituitary adenoma and 7 were atypical pituitary adenoma. Of the atypical pituitary adenoma cases, one case was ACTH, one GH and one both GH and prolactin hormone immunoreactive pituitary adenomas. Four cases were hormone immunonegative adenomas. Of the typical pituitary adenoma cases, 39 cases were GH, 19 ACTH, 17 prolactin, 10 FSH, 8 LH and one TSH immunreactive pituitary adenomas. Fourty-seven cases were hormone immunonegative adenomas.Twenty-two of the all pitutary adenoma cases had recurrence. Of these cases, 18 were typical adenoma and four were atypical adenoma.Conclusion: The ratio of prolactin immunoreactive pituitary adenoma cases in the surgical material of neuropathology is decreasing due to medical therapy. Atypical pituitary adenomas are not the sole factor affecting the recurrence mechanism but these tumors have higher recurrence rate compared with typical pituitary adenomas and we think the proliferation index might be the principal approach in the diagnosis of these lesions.

  18. Identification and differential distribution of CART in the small intestine depending on the diet. (United States)

    Janiuk, I; Olkowski, B; Szczotka-Bochniarz, A


    This study was aimed at identifying and locating cocaine- and amphetamine-regulated transcript (CART) in the small intestine of broilers in relation to the diet. The feeding regime of the chicks was based on diets largely consisting of maize and one of four protein sources: post-extraction soya bean meal (SBM) or non-GM seed meal - meal from traditional variety of soy seeds Glicine max (FFS) and meal from seeds of Lupinus angustifolius (LA) and Lupinus luteus L (LY). The presence of cocaine- and amphetamine-regulated transcript immunoreactive (CART-IR) in the wall of the small intestine of the chicks was determined on the basis of staining patterns produced by the immunohistochemical method (IHC). CART-IR structures were found in the myenteric plexus (MP), submucosus plexus (SP), in endomucosal fibres, and fibres innervating miocytes and blood vessels in the muscularis membrane and adipocytes of the white adipose tissue (WAT) located on the perimeter of the serous membrane and single cells of the diffuse neuroendocrine system. Based on microscopic observation and result analysis, the lowest number of CART-IR structures was identified in the group that was fed the SBM-based diet. This study confirms previous observations concerning CART distribution in the gastrointestinal tract (GIT) of animal and broadens current knowledge by inclusion of chicken in the list of CART-positive species. Moreover, this work provides evidence that dietary composition can be a factor that stimulates post-prandial CART secretion in intestinal nerve structures. PMID:24797515

  19. Comparison of catalase immunoreactivity in the hippocampus between young, adult and aged mice and rats. (United States)

    Ahn, Ji Hyeon; Chen, Bai Hui; Shin, Bich-Na; Lee, Tae-Kyeong; Cho, Jeong Hwi; Kim, In Hye; Park, Joon Ha; Lee, Jae-Chul; Tae, Hyun-Jin; Lee, Choong-Hyun; Won, Moo-Ho; Lee, Yun Lyul; Choi, Soo Young; Hong, Seongkweon


    Catalase (CAT) is an important antioxidant enzyme and is crucial in modulating synaptic plasticity in the brain. In this study, CAT expression as well as neuronal distribution was compared in the hippocampus among young, adult and aged mice and rats. Male ICR mice and Sprague Dawley rats were used at postnatal month (PM) 1, PM 6 and PM 24 as the young, adult and aged groups, respectively (n=14/group). CAT expression was examined by immunohistochemistry and western blot analysis. In addition, neuronal distribution was examined by NeuN immunohistochemistry. In the present study, the mean number of NeuN‑immunoreactive neurons was marginally decreased in mouse and rat hippocampi during aging, although this change was not identified to be significantly different. However, CAT immunoreactivity was significantly increased in pyramidal and granule neurons in the adult mouse and rat hippocampi and was significantly decreased in the aged mouse and rat hippocampi compared with that in the young animals. CAT protein levels in the hippocampus were also lowest in the aged mouse and rat hippocampus. These results indicate that CAT expression is significantly decreased in the hippocampi of aged animals and decreased CAT expression may be closely associated with aging. PMID:27221506

  20. Genetically engineered alginate lyase-PEG conjugates exhibit enhanced catalytic function and reduced immunoreactivity.

    Directory of Open Access Journals (Sweden)

    John W Lamppa

    Full Text Available Alginate lyase enzymes represent prospective biotherapeutic agents for treating bacterial infections, particularly in the cystic fibrosis airway. To effectively deimmunize one therapeutic candidate while maintaining high level catalytic proficiency, a combined genetic engineering-PEGylation strategy was implemented. Rationally designed, site-specific PEGylation variants were constructed by orthogonal maleimide-thiol coupling chemistry. In contrast to random PEGylation of the enzyme by NHS-ester mediated chemistry, controlled mono-PEGylation of A1-III alginate lyase produced a conjugate that maintained wild type levels of activity towards a model substrate. Significantly, the PEGylated variant exhibited enhanced solution phase kinetics with bacterial alginate, the ultimate therapeutic target. The immunoreactivity of the PEGylated enzyme was compared to a wild type control using in vitro binding studies with both enzyme-specific antibodies, from immunized New Zealand white rabbits, and a single chain antibody library, derived from a human volunteer. In both cases, the PEGylated enzyme was found to be substantially less immunoreactive. Underscoring the enzyme's potential for practical utility, >90% of adherent, mucoid, Pseudomonas aeruginosa biofilms were removed from abiotic surfaces following a one hour treatment with the PEGylated variant, whereas the wild type enzyme removed only 75% of biofilms in parallel studies. In aggregate, these results demonstrate that site-specific mono-PEGylation of genetically engineered A1-III alginate lyase yielded an enzyme with enhanced performance relative to therapeutically relevant metrics.

  1. Light microscopic image analysis system to quantify immunoreactive terminal area apposed to nerve cells (United States)

    Wu, L. C.; D'Amelio, F.; Fox, R. A.; Polyakov, I.; Daunton, N. G.


    The present report describes a desktop computer-based method for the quantitative assessment of the area occupied by immunoreactive terminals in close apposition to nerve cells in relation to the perimeter of the cell soma. This method is based on Fast Fourier Transform (FFT) routines incorporated in NIH-Image public domain software. Pyramidal cells of layer V of the somatosensory cortex outlined by GABA immunolabeled terminals were chosen for our analysis. A Leitz Diaplan light microscope was employed for the visualization of the sections. A Sierra Scientific Model 4030 CCD camera was used to capture the images into a Macintosh Centris 650 computer. After preprocessing, filtering was performed on the power spectrum in the frequency domain produced by the FFT operation. An inverse FFT with filter procedure was employed to restore the images to the spatial domain. Pasting of the original image to the transformed one using a Boolean logic operation called 'AND'ing produced an image with the terminals enhanced. This procedure allowed the creation of a binary image using a well-defined threshold of 128. Thus, the terminal area appears in black against a white background. This methodology provides an objective means of measurement of area by counting the total number of pixels occupied by immunoreactive terminals in light microscopic sections in which the difficulties of labeling intensity, size, shape and numerical density of terminals are avoided.

  2. Increases in Doublecortin Immunoreactivity in the Dentate Gyrus following Extinction of Heroin-Seeking Behavior

    Directory of Open Access Journals (Sweden)

    Megan P. Hicks


    Full Text Available Adult-generated neurons in the dentate gyrus (DG of the hippocampus play a role in various forms of learning and memory. However, adult born neurons in the DG, while still at an immature stage, exhibit unique electrophysiological properties and are also functionally implicated in learning and memory processes. We investigated the effects of extinction of drug-seeking behavior on the formation of immature neurons in the DG as assessed by quantification of doublecortin (DCX immunoreactivity. Rats were allowed to self-administer heroin (0.03 mg/kg/infusion for 12 days and then subjected either to 10 days of extinction training or forced abstinence. We also examined extinction responding patterns following heroin self-administration in glial fibrillary acidic protein thymidine kinase (GFAP-tk transgenic mice, which have been previously demonstrated to show reduced formation of immature and mature neurons in the DG following treatment with ganciclovir (GCV. We found that extinction training increased DCX immunoreactivity in the dorsal DG as compared with animals undergoing forced abstinence, and that GCV-treated GFAP-tk mice displayed impaired extinction learning as compared to saline-treated mice. Our results suggest that extinction of drug-seeking behavior increases the formation of immature neurons in the DG and that these neurons may play a functional role in extinction learning.

  3. Structural changes in emulsion-bound bovine beta-lactoglobulin affect its proteolysis and immunoreactivity. (United States)

    Marengo, Mauro; Miriani, Matteo; Ferranti, Pasquale; Bonomi, Francesco; Iametti, Stefania; Barbiroli, Alberto


    Adsorption on the surface of sub-micrometric oil droplets resulted in significant changes in the tertiary structure of bovine beta-lactoglobulin (BLG), a whey protein broadly used as a food ingredient and a major food allergen. The adsorbed protein had increased sensitivity to trypsin, and increased immunoreactivity towards specific monoclonal antibodies. In spite of the extensive tryptic breakdown of emulsion-bound BLG, some sequence stretches in BLG became trypsin-insensitive upon absorption of the protein on the fat droplets. As a consequence - at contrast with free BLG - proteolysis of emulsion-bound BLG did not decrease the immunoreactivity of the protein, and some of the large peptides generated by trypsinolysis of emulsion-bound BLG were still recognizable by specific monoclonal antibodies. Structural changes occurring in emulsion-bound BLG and their consequences are discussed in comparison with those occurring when the tertiary structure of BLG is modified by lipophilic salts, by urea, or upon interaction with solid hydrophobic surfaces. Such a comparison highlights the relevance of situation-specific structural modifications, that in turn may affect physiologically relevant features of the protein. PMID:27085639

  4. HNK-1 immunoreactivity during early morphogenesis of the head region in a nonmodel vertebrate, crocodile embryo (United States)

    Kundrát, Martin


    The present study examines HNK-1 immunoidentification of a population of the neural crest (NC) during early head morphogenesis in the nonmodel vertebrate, the crocodile ( Crocodylus niloticus) embryos. Although HNK-1 is not an exclusive NC marker among vertebrates, temporospatial immunoreactive patterns found in the crocodile are almost consistent with NC patterns derived from gene expression studies known in birds (the closest living relatives of crocodiles) and mammals. In contrast to birds, the HNK-1 epitope is immunoreactive in NC cells at the neural fold level in crocodile embryos and therefore provides sufficient base to assess early migratory events of the cephalic NC. I found that crocodile NC forms three classic migratory pathways in the head: mandibular, hyoid, and branchial. Further, I demonstrate that, besides this classic phenotype, there is also a forebrain-derived migratory population, which consolidates into a premandibular stream in the crocodile. In contrast to the closely related chick model, crocodilian premandibular and mandibular NC cells arise from the open neural tube suggesting that species-specific heterochronic behavior of NC may be involved in the formation of different vertebrate facial phenotypes.

  5. Limited hydrolysis combined with controlled Maillard-induced glycation does not reduce immunoreactivity of soy protein for all sera tested. (United States)

    Walter, Jordan; Greenberg, Yana; Sriramarao, P; Ismail, Baraem P


    Combining proteolysis and Maillard-induced glycation was investigated to reduce the immunoreactivity of soy protein. Soy protein was hydrolyzed by Alcalase following response surface methodology utilizing three variables, temperature, time, and enzyme:substrate ratio, with the degree of hydrolysis (DH) and percent reduction in immunoreactivity as response variables. Western blots and ELISA were used to evaluate immunoreactivity using human sera. Data were fitted to appropriate models and prediction equations were generated to determine optimal hydrolysis conditions. The hydrolysate produced under optimized conditions was subjected to glycation with dextran. Hydrolysate produced under optimal conditions had 7.8% DH and a percent reduction in immunoreactivity ranging from 20% to 52%, depending on the sera used. Upon glycation, immunoreactivity was further reduced only when using serum that had the highest soy-specific IgE. This work revealed limitations and provided premises for future studies intended to prove the potency of the combined modification approach to produce a hypoallergenic protein ingredient. PMID:27451243

  6. Immunohistochemical detection of ganglia in the rat stomach serosa, containing neurons immunoreactive for gastrin-releasing peptide and vasoactive intestinal peptide

    DEFF Research Database (Denmark)

    Poulsen, Steen Seier; Holst, J J


    Ganglia, not previously described, were identified in the rat stomach serosa along the minor curvature. The ganglia consisted of varying number of cell bodies lying in clusters along or within nerve bundles. The ganglia were shown to contain GRP and VIP immunoreactive nerve fibers and cell bodies...... and also some NPY immunoreactive fibers, whereas they were devoid of somatostatin immunoreactivity. Nerve ligation experiments indicated that the ganglia are intrinsic to the stomach....

  7. Intraepidermal neuron-specific enolase (NSE)-immunoreactive nerve fibres: evidence for sprouting in uremic patients on maintenance hemodialysis. (United States)

    Johansson, O; Hilliges, M; Ståhle-Bäckdahl, M


    The use of indirect immunohistochemistry in 12 patients on maintenance hemodialysis has shown weak or moderately strong neuron-specific enolase (NSE)-immunoreactive nerve terminals and fibres sprouting throughout the layers of the epidermis. No such terminals or fibres were found in any of 15 controls. There was no difference between uremic patients with pruritus and those without. Furthermore, NSE-positive nerve fibres with a normal appearance were seen in the dermis, at the epidermal-dermal junctional zone and sometimes entering the stratum basale in both patients and controls. The immunoreactive nerves were thin, smooth and, at their terminal fields, varicose. The immunoreactivity seemed to be associated chiefly with sensory nerves. Thus, our results suggest that uremic patients undergoing maintenance hemodialysis develop an abnormal pattern of cutaneous innervation. PMID:2657508

  8. Neuropeptide Y-like immunoreactive neurons in the suprachiasmatic-subparaventricular region in the hedgehog-tenrec. (United States)

    Künzle, H; Unger, J W


    The distribution of the neuropeptide Y (NPY) was studied in geniculate and peri-chiasmatic regions in the lesser hedgehog-tenrec, Echinops telfairi (Insectivora). Only few neurons demonstrated NPY-like immunoreactivity in the ventral lateral geniculate nucleus. In contrast, NPY-immunoreactive perikarya were clearly present in the suprachiasmatic nucleus (SCh) and dorsal and caudal to it. The latter region might correspond to the subparaventricular zone (SPV), recently identified in the rat as an additional area involved in processing circadian rhythms. While the distribution of a distinct cell population across nuclear boundries in both SCh and SPV might conform to the present idea of processing circadian rhythms, the presence of NPY-like immunoreactive neurons in these areas is rather unusual. In mammals, such neurons have only been demonstrated so far in the mentioned insectivore as well as in man. PMID:1515927

  9. Glucose transporter 5 (GLUT5)-like immunoreactivity is localized in subsets of neurons and glia in the rat brain. (United States)

    Kojo, Akiko; Yamada, Kentaro; Yamamoto, Toshiharu


    This study aimed at examining the distribution of glucose transporter 5 (GLUT5), which preferentially transports fructose, in the rat brain by immunohistochemistry and Western blotting. Small immunoreactive puncta (less than 0.7μm) were sparsely distributed all over the brain, some of which appeared to be associated with microglial processes detected by an anti-ionized calcium-binding adapter molecule 1 (Iba-1) monoclonal antibody. In addition, some of these immunoreactive puncta seemed to be associated with tanycyte processes that were labeled with anti-glial fibrillary acidic protein (GFAP) monoclonal antibody. Ependymal cells were also found to be immunopositive for GLUT5. Furthermore, several noticeable GLUT5 immunoreactive profiles were observed. GLUT5 immunoreactive neurons, confirmed by double staining with neuronal nuclei (NeuN), were seen in the entopeduncular nucleus and lateral hypothalamus. Cerebellar Purkinje cells were immunopositve for GLUT5. Dense accumulation of immunoreactive puncta, some of which were neuronal elements (confirmed by immunoelectron microscopy), were observed in the optic tract and their terminal fields, namely, superior colliculus, pretectum, nucleus of the optic tract, and medial terminal nucleus of the optic tract. In addition to the associated areas of the visual system, the vestibular and cochlear nuclei also contained dense GLUT5 immunoreactive puncta. Western blot analysis of the cerebellum indicated that the antibody used recognized the 33.5 and 37.0kDa bands that were also contained in jejunum and kidney extracts. Thus, these results suggest that GLUT5 may transport fructose in subsets of the glia and neurons for an energy source of these cells. PMID:27036089

  10. Role of neuropsin in parvalbumin immunoreactivity changes in hippocampal basket terminals of mice reared in various environments

    Directory of Open Access Journals (Sweden)

    Harumitsu eSuzuki


    Full Text Available In vitro approaches have suggested that neuropsin (or kallikrein 8/KLK8, which controls gamma-aminobutyric acid (GABA neurotransmission through neuregulin-1 and its receptor (ErbB4, is involved in neural plasticity (Tamura et al., 2012, 2013. In the present study, we examined whether parvalbumin (PV-positive neuronal networks, the majority of which are ErbB4-positive GABAergic interneurons, are controlled by neuropsin in tranquil and stimulated voluntarily behaving mice.PV-immunoreactive fibers surrounding hippocampal pyramidal and granular neurons in mice reared in their home cage were decreased in neuropsin-deficient mice, suggesting that neuropsin controls PV immunoreactivity. One- or two-week exposures of wild mice to novel environments, in which they could behave freely and run voluntarily in a wheel resulted in a marked upregulation of both neuropsin mRNA and protein in the hippocampus. To elucidate the functional relevance of the increase in neuropsin during exposure to a rich environment, the intensities of PV-immunoreactive fibers were compared between neuropsin-deficient and wild-type mice under environmental stimuli. When mice were transferred into novel cages (large cages with toys, the intensity of PV-immunoreactive fibers increased in wild-type mice and neuropsin-deficient mice. Therefore, behavioral stimuli control a neuropsin-independent form of PV immunoreactivity. However, the neuropsin-dependent part of the change in PV-immunoreactive fibers may occur in the stimulated hippocampus because increased levels of neuropsin continued during these enriched conditions.

  11. Homogeneous MGMT immunoreactivity correlates with an unmethylated MGMT promoter status in brain metastases of various solid tumors.

    Directory of Open Access Journals (Sweden)

    Barbara Ingold

    Full Text Available The O(6-methylguanine-methyltransferase (MGMT promoter methylation status is a predictive parameter for the response of malignant gliomas to alkylating agents such as temozolomide. First clinical reports on treating brain metastases with temozolomide describe varying effects. This may be due to the fact that MGMT promoter methylation of brain metastases has not yet been explored in depth. Therefore, we assessed MGMT promoter methylation of various brain metastases including those derived from lung (n = 91, breast (n = 72 kidney (n = 49 and from malignant melanomas (n = 113 by methylation-specific polymerase chain reaction (MS-PCR and MGMT immunoreactivity. Fifty-nine of 199 brain metastases (29.6% revealed a methylated MGMT promoter. The methylation rate was the highest in brain metastases derived from lung carcinomas (46.5% followed by those from breast carcinoma (28.8%, malignant melanoma (24.7% and from renal carcinoma (20%. A significant correlation of homogeneous MGMT-immunoreactivity (>95% MGMT positive tumor cells and an unmethylated MGMT promoter was found. Promoter methylation was detected in 26 of 61 (43% tumors lacking MGMT immunoreactivity, in 17 of 63 (27% metastases with heterogeneous MGMT expression, but only in 5 of 54 brain metastases (9% showing a homogeneous MGMT immunoreactivity. Our results demonstrate that a significant number of brain metastases reveal a methylated MGMT-promoter. Based on an obvious correlation between homogeneous MGMT immunoreactivity and unmethylated MGMT promoter, we hypothesize that immunohistochemistry for MGMT may be a helpful diagnostic tool to identify those tumors that probably will not benefit from the use of alkylating agents. The discrepancy between promoter methylation and a lack of MGMT immunoreactivity argues for assessing MGMT promoter methylation both by immunohistochemical as well as by molecular approaches for diagnostic purposes.

  12. Effects of Static Magnetic Field on Growth of Leptospire, Leptospira interrogans serovar canicola: Immunoreactivity and Cell Division

    CERN Document Server

    Triampo, W; Triampo, D; Wong-Ekkabut, J; Tang, I M; Triampo, Wannapong; Doungchawee, Galayanee; Triampo, Darapond; Wong-Ekkabut, Jirasak


    The effects of the exposure of the bacterium, Leptospira interrogans serovar canicola to a constant magnetic field with magnetic flux density from a permanent ferrite magnet = 140 mT were studied. Changes in Leptospira cells after their exposure to the field were determined on the basis of changes in their growth behavior and agglutination immunoreactivity with a homologous antiserum using darkfield microscopy together with visual imaging. The data showed that the exposed Leptospira cells have lower densities and lower agglutination immunoreactivity than the unexposed control group. Interestingly, some of the exposed Leptospira cells showed abnormal morphologies such as large lengths. We discussed some of the possible reasons for these observations.

  13. Distribution of obestatin and ghrelin in human tissues: immunoreactive cells in the gastrointestinal tract, pancreas, and mammary glands

    DEFF Research Database (Denmark)

    Grönberg, Malin; Tsolakis, Apostolos V; Magnusson, Linda;


    specifically recognizes human obestatin was produced. Using this antibody and a commercial antibody vs ghrelin, the distribution of obestatin and ghrelin immunoreactive cells was determined in a panel of human tissues using immunohistochemistry. The two peptides were detected in the mucosa of the......Obestatin and ghrelin are two peptides derived from the same prohormone. It is well established that ghrelin is produced by endocrine cells in the gastric mucosa. However, the distribution of human obestatin immunoreactive cells is not thoroughly characterized. A polyclonal antibody that...

  14. Immunoreactive transforming growth factor alpha and epidermal growth factor in oral squamous cell carcinomas

    DEFF Research Database (Denmark)

    Therkildsen, M H; Poulsen, Steen Seier; Bretlau, P


    Forty oral squamous cell carcinomas have been investigated immunohistochemically for the presence of transforming growth factor alpha (TGF-alpha) and epidermal growth factor (EGF). The same cases were recently characterized for the expression of EGF-receptors. TGF-alpha was detected with a...... monoclonal mouse antibody and EGF with polyclonal rabbit antiserum. Thirty-five of the tumours were positive for TGF-alpha and 26 of the tumours for EGF. None of the poorly differentiated tumours was positive for EGF, but they all were for TGF-alpha. In sections including normal differentiated oral mucosa......, the cells above the basal cell layer were positive for both TGF-alpha and EGF. The same staining pattern was observed in oral mucosa obtained from healthy persons. In moderately to well differentiated carcinomas, the immunoreactivity was mainly confined to the cytologically more differentiated cells...

  15. Immunoreactivity and Radioimmunoscintigraphy of 4-Lysine Single Chain (Fv) Lym-1 Antibody for the Radiometal Chelation

    International Nuclear Information System (INIS)

    Small size of recombinant scFv, composed of VH and VL region of IgG, has many advantages such as faster blood clearance, improved tumor localization and reduced human anti-mouse antibody (HAMA) response. On the other hand, owing to small size, number of amino group, which was not involved in binding site, of ScFv lym-1 was insufficient in conjugation with CITC-DTPA chelator for radio metal labeling. The goal of this study is to introduce 4-lysine tag to the end of ScFv lym-1 sequence for radio metal conjugation and to evaluate the immunoreactivity and radioimmunoscintigraphy of chelator conjugated 4-lysine taq scFv lym-1 (4-lys scFv)

  16. Human papillomavirus and p53 protein immunoreactivity in condylomata acuminatum and squamous cell carcinoma of penis

    Institute of Scientific and Technical Information of China (English)

    Xin-Hua ZHANG; Gui-Qin SUN; Yu YANG; Tai-He ZHANG


    To determine the immunoreactive pattem of human papillomavirus (HPV) antigen and p53 protein in condylomata acuminatum (CA) and squamous cell carcinoma (SCC) of penis. Methods: Immunohistochemistry for HPV and p53 were performed in 40 specimens of formalin fixed, paraffin embedded tissues using a polyclonal (rabbit) antibody against HPV and a monoclonal (mouse) antibody against human p53 protein. Twenty one cases of CA and nineteen cases of SCC were examined. Results: HPV antigen was detected in all 21 CA and 2 penile SCC. p53 protein overexpression was observed in 12 of 19 (63%) SCC in which 6 cases were strong positive. Five of 21 CA (24%)showed low-grade p53 protein overexpression. Conclusion: CA is related to HPV infection and some cases show p53 protein low-grade overexpression. In contrast, p53 protein overexpression is common in penile SCC, which is seldom related to HPV infection.

  17. Immunoreactive luteinizing hormone-releasing hormone in the seminal plasma and human semen parameters

    International Nuclear Information System (INIS)

    A luteinizing hormone-releasing hormone (LH-RH)-like substance has been detected in human seminal plasma by a radioimmunoassay (RIA) with a highly specific anti-LH-RH antiserum. The seminal samples - not only the plasma itself but also the sample extracted by an acid/alcohol method - showed satisfactory displacement curves in our RIA system. The relationship between fertility and the LH-RH values in the seminal plasma was studied by comparing the peptide levels with sperm concentration and motility. By these two parameters, 103 samples were divided into four groups. In the low-concentration groups (oligozoospermic patients), the hormonal concentrations differed significantly between those specimens demonstrating good and poor motility. These data suggest that this immunoreactive LH-RH may play a role in human spermatogenesis

  18. Tau immunoreactivity detected in human plasma, but no obvious increase in dementia

    DEFF Research Database (Denmark)

    Ingelson, M; Blomberg, M; Benedikz, Eirikur;


    Tau proteins are central to the neuropathology of Alzheimer's disease and tau levels in cerebrospinal fluid are elevated in affected individuals. In this study, we investigated the presence of tau in plasma from subjects with Alzheimer's disease (n = 16), frontotemporal dementia (n = 10), vascular...... dementia (n = 16) and from healthy controls (n = 15). By using an ELISA with monoclonal tau antibodies, tau immunoreactivity was detected in approximately 20% of the subjects. However, no difference between the disease and control groups was seen. After gel filtration of tau immunopositive plasma, the peak...... reactivity was found in the 160-kD fraction, indicating the source to be tau-like molecules of high-molecular-weight or polymers of low-molecular-weight tau isoforms. We conclude that measurements of tau in plasma cannot be utilized diagnostically for Alzheimer's disease or for the other dementias...

  19. Effects of mutations on enzyme activity and immunoreactivity of the S1 subunit of pertussis toxin. (United States)

    Lobet, Y; Cieplak, W; Smith, S G; Keith, J M


    By introducing a series of six different substitutions at and around position 9, we investigated the structural requirements of the amino-terminal region of the S1 subunit of pertussis toxin for both enzyme activity and immunoreactivity. All mutant S1 analogs with a substitution at this location exhibited severely decreased ADP-ribosyltransferase activity (range, 400- to 2,500-fold). In contrast, alteration of arginine 58 had considerably less effect. The reactivity of the mutant molecules with monoclonal antibody 1B7 varied with the nature of the substitution. These findings indicate an absolute requirement for the presence of an arginine residue at position 9 for the maintenance of efficient ADP-ribosyltransferase activity and illustrate the specific participation of vicinal residues in the formation of the protective epitope. PMID:2807541

  20. Oxidation of proline decreases immunoreactivity and alters structure of barley prolamin. (United States)

    Huang, Xin; Sontag-Strohm, Tuula; Stoddard, Frederick L; Kato, Yoji


    Elimination of celiac-toxic prolamin peptides and proteins is essential for Triticeae products to be gluten-free. Instead of enzymatic hydrolysis, in this study we investigated metal-catalyzed oxidation of two model peptides, QQPFP, and PQPQLPY, together with a hordein isolate from barley (Hordeum vulgare L.). We established a multiple reaction monitoring (MRM) LC-MS method to detect and quantify proline oxidation fragments. In addition to fragmentation, aggregation and side chain modifications were identified, including free thiol loss, carbonyl formation, and dityrosine formation. The immunoreactivity of the oxidized hordein isolate was considerably decreased in all metal-catalyzed oxidation systems. Cleavage of peptides or protein fragments at the numerous proline residues partially accounts for the decrease. Metal-catalyzed oxidation can thus be used in the modification and elimination of celiac-toxic peptides and proteins. PMID:27507515

  1. Reduction of immunoreactivity of bovine beta-lactoglobulin upon combined physical and proteolytic treatment

    DEFF Research Database (Denmark)

    Bonomi, F.; Fiocchi, A.; Frøkiær, Hanne; Gaiaschi, A.; Iametti, S.; Poiesi, C.; Rasmussen, P.; Restani, P.; Rovere, P.


    Bovine beta-lactoglobulin was hydrolyzed with trypsin or chymotrypsin before, during and after treatment at 600 MPa and pH 6.8 for 10 min at 30, 37 and 44degreesC. The extent of beta-lactoglobulin hydrolysis under pressure was noticeably higher than at atmospheric pressure, particularly when...... from those obtained at atmospheric pressure when chymotrypsin was used. The residual immunochemical reactivity of the products of combined pressure-enzyme treatment was assessed on the unresolved hydrolysates by ELISA tests using polyclonal and monoclonal antibodies, and on individual hydrolytic...... fractions by Western Blotting using sera of paediatric patients allergic to whey proteins in cow milk. The immunoreactivity of the whole hydrolysates was related to their content of residual intact beta-lactoglobulin, and no immunochemical reactivity was found for all the products of chymotrypsin hydrolysis...

  2. Characterisation of recombinant immunoreactive antigens of the scab mite Sarcoptes scabiei. (United States)

    Kuhn, C; Lucius, R; Matthes, H F; Meusel, G; Reich, B; Kalinna, B H


    Sarcoptic mange (or scabies) is an important skin disease which can affect a variety of species including humans, cattle, goats, sheep, horses, pigs, rabbits, and dogs. Approximately 300 million people are affected worldwide and in lifestock animals the infestation may lead to substantial economic losses caused by depression in growth and feed conversion rates. Diagnosis of Sarcoptes infestation is difficult and only a few serological tests have been developed using whole mite antigen for diagnosis of mange in animals. Here we describe the isolation and characterisation of cDNAs of several immunoreactive clones and their recombinant expression in Escherichia coli. Three of the proteins contain repetitive sequences which suggests that they might be involved in immune evasion. The application of these antigens in serodiagnosis and the suitability for diagnosis is discussed. PMID:18359167

  3. Variable sequences in a mosaic-like domain of meningococcal tbp2 encode immunoreactive epitopes. (United States)

    Rokbi, B; Maitre-Wilmotte, G; Mazarin, V; Fourrichon, L; Lissolo, L; Quentin-Millet, M J


    Transferrin-binding proteins from Neisseria meningitidis vary among different isolates. We have identified and studied a hypervariable region adjacent to the carboxyl-end of the transferrin-binding domain of the Tbp2 molecule. The tbp2 genes from six strains of N. meningitidis were cloned and sequenced in this particular region. Sequence analysis of these regions along with five other sequences available from pathogenic Neisseria showed a common organisation of seven highly variable nucleotide stretches interspersed with six conserved nucleotide stretches. The variable regions correlated with the location of immunoreactive epitopes in polyclonal antisera raised to transferrin-binding proteins identified by peptide pin technology. Sequence analysis suggested a mosaic-like organisation of the tbp2 genes. Taken together, these data suggest that the antigenic variation in this part of the protein may result from a strong host immune pressure. PMID:7590185

  4. Beta-endorphin immunoreactivity during high-intensity exercise with and without opiate blockade. (United States)

    Angelopoulos, T J


    Nine highly fit men [mean (SE) maximum oxygen uptake, VO2max: 63.9 (1.7) ml x kg(-1) x min(-1); age 27.6 (1.6) years] were studied during two treadmill exercise trials to determine plasma beta-endorphin immunoreactivity during intense exercise (80% VO2max). A double-blind experimental design was used, and subjects performed the two exercise trials in counterbalanced order. Exercise trials were 30 min in duration and were conducted 7 days apart. One exercise trial was undertaken following administration of naloxone (1.2 3 cm3) and the other after receiving a placebo (0.9% NaCl saline; 3 cm3). Prior to each experimental trial, a flexible catheter was placed into an antecubital vein and baseline blood samples were collected. Thereafter, each subject received either a naloxone or placebo bolus injection. Blood samples were also collected after 10, 20 and 30 min of continuous exercise. beta-Endorphin was higher (P exercise when compared to pre-exercise in both trials. However, no statistically significant difference was found (P> 0.05) between exercise time points within either experimental trial. beta-endorphin immunoreactivity was greater (P exercise sampling time point [10 min: 63.7 (3.9) pg x ml(-1) vs 78.7 (3.8) pg x ml(-1); 20 min: 68.7 (4.1) pg x ml(-1) vs (4.3) pg x ml(-1); 30 min: 71.0 (4.3) pg x ml(-1) vs 82.5(3.2) pg x ml(-1)]. These data suggest that intense exer induces significant increases in beta-endorphin that are maintained over time during steady-rate exercise. Exercise and naloxone had an interactive effect on beta-endorphin release that warrants further investigation. PMID:11820329

  5. Neurotrophin-receptor immunoreactive neurons in mesopontine regions involved in the control of behavioral states. (United States)

    Yamuy, J; Sampogna, S; Chase, M H


    The microinjection of nerve growth factor (NGF) and neurotrophin-3 (NT-3) into the rostral pontine tegmentum of adult cats rapidly induces long-lasting episodes of rapid eye movement (REM) sleep [J. Yamuy, F.R. Morales, M.H. Chase, Induction of rapid eye movement sleep by the microinjection of nerve growth factor into the pontine reticular formation of the cat, Neuroscience 66 (1995) 9-13]. Because this effect may be mediated by neurotrophin receptors, we sought to determine the distribution of neurons that contain low- and high-affinity neurotrophin receptors in regions of the feline pons and mesencephalon which are involved in the generation of REM sleep as well as neuronal groups that are involved in the control of REM sleep-related patterns of physiological activity. Using antibodies directed against p75, trkA, trkB and trkC, immunolabeled neurons were present in the latero-dorsal and pedunculo-pontine tegmental nuclei, the peribrachial nuclei, medial and lateral pontine reticular formation, the raphe nuclei, and the locus coeruleus. Giant reticular cells and large neurons in the mesencephalic trigeminal nucleus were immunoreactive for p75 and all trk receptors. Neurons that were devoid of neurotrophin-receptor immunoreactivity were intermingled with immunostained neurons in all explored structures. Thus, both low- and high-affinity neurotrophin receptors are conspicuously present in neurons located in mesopontine regions of adult cats. These data underscore the importance of neurotrophin-induced trophic actions on mesopontine neurons. Furthermore, the results support the hypothesis that NGF and NT-3 may modulate the electrical activity of neurons in the rostral pontine tegmentum that are responsible for the generation of REM sleep by acting on one or more of the neurotrophin receptors. PMID:10825475

  6. Advantage of highly immunoreactive monoclonal antibodies in radioimmunoscintigraphy for tumor detection, (2)

    International Nuclear Information System (INIS)

    There is theoretically a potential benefit in using a highly immunoreactive monoclonal antibody. The effect of immunoreactivity (IR) on the antibody biodistribution, however, has not yet been described in detail. Thus, this study was designed to investigate the effect of IR on the biodistribution in an animal model. The hydroxylapatite high performance liquid chromatography (HA-HPLC) system has been tested and confirmed to separate the F ab 96.5, an anti melanoma p97 antigen, into high and low IR fractions. 125I-F ab 96.5 preparations with a different IR were administered to groups of nude mice bearing FEM-XII human skin melanoma xenografts for biodistribution and imaging studies. The biodistribution data showed that the high IR antibody improved tumor targeting by increasing activity ratios of tumor to non tumor tissue; the mechanism for the increased tumor to non tumor ratios was increased tumor activity uptake and prolonged tumor activity retention with associated rapid clearance from the blood and non tumor sites. The imaging study visually supported the results obtained in the biodistribution study; the high IR antibody demonstrated better and earlier tumor delineation and the tumor to non tumor contrast continued to improve with time. In this model system, where the whole body clearance rate was the same for the high IR and low IR preparations, the overall antibody metabolism and excretion were not significantly dependent on IR. Therefore, the effect of IR is to alter the distribution of antibody between tumor and blood, with high IR having increased tumor activity and reduced blood activity (consequently reduced non tumor organ activity). This would also be beneficial for therapeutic use of radiolabeled antibodies, since high IR antibodies can minimize undesirable radiation exposure to normal organs. In conclusion, high IR antibodies are essential for optimal tumor targeting. (author)

  7. Production of the recombinant single chain anti-B cell lymphoma antibody and evaluation of immunoreactivity

    International Nuclear Information System (INIS)

    Recombinant ScFv lym-1 was produced, using pET vector system for large scale production. ScFv lym-1 gene inserted pET-22b (+) vector, was expressed in E. coli BL-21 strain. ScFv lym-1 antibody extracted from periplasm, was purified with His-Taq column. To evaluated immunoreactivity with Raji cell, ScFv lym-1 was labeled with I-125 and I-125 ScFv lym-1 was purified with desalting column. Raji cell was injected into the C57BR/cdJ SCID mice. Gamma camera imaging were taken time point at 1, 8, 24 and 48 hr with 8 mm pinhole collimator. An active scFv lym-1 could be produced in E. coli with soluble from using pET vector system. Immunoreactivity and affinity constant of lgG lym-1 were 54% and 1.83 x 109 M-1, respectively, and those of scFv lym-1 were 53.7% and 1.46 x 109 M-1, respectively. Biodistribution of I-125 scFv lym-1 antibody showed faster clearance in blood, spleen, kidney and than I-125 lgG lym-1 antibody. Gamma camera image of I-125 scFv lym-1 antibody showed faster clearance and tumor targeting liver than I-125 lgG lym-1 antibody. In vitro properties of scFv lym-1 were similar to those of lgG lym-1. ScFv lym-1 showed faster blood clearance than lgG lym-1. These results suggest that scFv lym-1 antibody can be useful for tumor imaging agent

  8. Substance P immunoreactivity exhibits frequent colocalization with kisspeptin and neurokinin B in the human infundibular region.

    Directory of Open Access Journals (Sweden)

    Erik Hrabovszky

    Full Text Available Neurons synthesizing neurokinin B (NKB and kisspeptin (KP in the hypothalamic arcuate nucleus represent important upstream regulators of pulsatile gonadotropin-releasing hormone (GnRH neurosecretion. In search of neuropeptides co-expressed in analogous neurons of the human infundibular nucleus (Inf, we have carried out immunohistochemical studies of the tachykinin peptide Substance P (SP in autopsy samples from men (21-78 years and postmenopausal (53-83 years women. Significantly higher numbers of SP-immunoreactive (IR neurons and darker labeling were observed in the Inf of postmenopausal women than in age-matched men. Triple-immunofluorescent studies localized SP immunoreactivity to considerable subsets of KP-IR and NKB-IR axons and perikarya in the infundibular region. In postmenopausal women, 25.1% of NKB-IR and 30.6% of KP-IR perikarya contained SP and 16.5% of all immunolabeled cell bodies were triple-labeled. Triple-, double- and single-labeled SP-IR axons innervated densely the portal capillaries of the infundibular stalk. In quadruple-labeled sections, these axons formed occasional contacts with GnRH-IR axons. Presence of SP in NKB and KP neurons increases the functional complexity of the putative pulse generator network. First, it is possible that SP modulates the effects of KP and NKB in axo-somatic and axo-dendritic afferents to GnRH neurons. Intrinsic SP may also affect the activity and/or neuropeptide release of NKB and KP neurons via autocrine/paracrine actions. In the infundibular stalk, SP may influence the KP and NKB secretory output via additional autocrine/paracrine mechanisms or regulate GnRH neurosecretion directly. Finally, possible co-release of SP with KP and NKB into the portal circulation could underlie further actions on adenohypophysial gonadotrophs.

  9. Effects of Dielectrophoresis on Growth, Viability and Immuno-reactivity of Listeria monocytogenes

    Directory of Open Access Journals (Sweden)

    Bhunia Arun K


    Full Text Available Abstract Dielectrophoresis (DEP has been regarded as a useful tool for manipulating biological cells prior to the detection of cells. Since DEP uses high AC electrical fields, it is important to examine whether these electrical fields in any way damage cells or affect their characteristics in subsequent analytical procedures. In this study, we investigated the effects of DEP manipulation on the characteristics of Listeria monocytogenes cells, including the immuno-reactivity to several Listeria-specific antibodies, the cell growth profile in liquid medium, and the cell viability on selective agar plates. It was found that a 1-h DEP treatment increased the cell immuno-reactivity to the commercial Listeria species-specific polyclonal antibodies (from KPL by ~31.8% and to the C11E9 monoclonal antibodies by ~82.9%, whereas no significant changes were observed with either anti-InlB or anti-ActA antibodies. A 1-h DEP treatment did not cause any change in the growth profile of Listeria in the low conductive growth medium (LCGM; however, prolonged treatments (4 h or greater caused significant delays in cell growth. The results of plating methods showed that a 4-h DEP treatment (5 MHz, 20 Vpp reduced the viable cell numbers by 56.8–89.7 %. These results indicated that DEP manipulation may or may not affect the final detection signal in immuno-based detection depending on the type of antigen-antibody reaction involved. However, prolonged DEP treatment for manipulating bacterial cells could produce negative effects on the cell detection by growth-based methods. Careful selection of DEP operation conditions could avoid or minimize negative effects on subsequent cell detection performance.

  10. Topography and time course of changes in spinal neuropeptide Y immunoreactivity after spared nerve injury. (United States)

    Intondi, A B; Zadina, J E; Zhang, X; Taylor, B K


    We used a new computer-assisted method to precisely localize and efficiently quantify increases in neuropeptide Y immunoreactivity (NPY-ir) along the mediolateral axis of the L4 dorsal horn (DH) following transection of either the tibial and common peroneal nerves (thus sparing the sural branch, spared nerve injury (SNI)), the tibial nerve, or the common peroneal and sural nerves. Two weeks after SNI, NPY-ir increased within the tibial and peroneal innervation territories; however, NPY-ir in the central-lateral region (innervated by the spared sural nerve) was indistinguishable from that of sham. Conversely, transection of the sural and common peroneal nerves induced an increase in NPY-ir in the central-lateral region, while leaving the medial region (innervated by the tibial nerve) unaffected. All nerve injuries increased NPY-ir in dorsal root ganglia (DRG) and nucleus gracilis (NG). By 24 weeks, both NPY-ir upregulation in the DH and hyper-responsivity to cold and noxious mechanical stimuli had resolved. Conversely, NPY-ir in DRG and NG, and hypersensitivity to non-noxious static mechanical stimuli, did not resolve within 24 weeks. Over this time course, the average cross-sectional area of NPY-immunoreactive DRG neurons increased by 151 mum(2). We conclude that the upregulation of NPY after SNI is restricted to medial zones of the DH, and therefore cannot act directly upon synapses within the more lateral (sural) zones to control sural nerve hypersensitivity. Instead, we suggest that NPY in the medial DH tonically inhibits hypersensitivity by interrupting mechanisms of central sensitization and integration of sensory signals at the spinal and supraspinal levels. PMID:19879928

  11. Comparative Mapping of GABA-Immunoreactive Neurons in the Buccal Ganglia of Nudipleura Molluscs. (United States)

    Gunaratne, Charuni A; Katz, Paul S


    Phylogenetic comparisons of neurotransmitter distribution are important for understanding the ground plan organization of nervous systems. This study describes the γ-aminobutyric acid (GABA)-immunoreactive (GABA-ir) neurons in the buccal ganglia of six sea slug species (Mollusca, Gastropoda, Euthyneura, Nudipleura). In the nudibranch species, Hermissenda crassicornis, Tritonia diomedea, Tochuina tetraquetra, and Dendronotus iris, the number of GABA-ir neurons was highly consistent. Another nudibranch, Melibe leonina, however, contained approximately half the number of GABA-ir neurons. This may relate to its loss of a radula and its unique feeding behavior. The GABA immunoreactivity in a sister group to the nudibranchs, Pleurobranchaea californica, differed drastically from that of the nudibranchs. Not only did it have significantly more GABA-ir neurons but it also had a unique GABA distribution pattern. Furthermore, unlike the nudibranchs, the Pleurobranchaea GABA distribution was also different from that of other, more distantly related, euopisthobranch and panpulmonate snails and slugs. This suggests that the Pleurobranchaea GABA distribution may be a derived feature, unique to this lineage. The majority of GABA-ir axons and neuropil in the Nudipleura were restricted to the buccal ganglia, commissures, and connectives. However, in Tritonia and Pleurobranchaea, we detected a few GABA-ir fibers in buccal nerves that innervate feeding muscles. Although the specific functions of the GABA-ir neurons in the species in this study are not known, the innervation pattern suggests these neurons may play an integrative or regulatory role in bilaterally coordinated behaviors in the Nudipleura. PMID:26355705

  12. Decreased nucleotide excision repair in steatotic livers associates with myeloperoxidase-immunoreactivity

    International Nuclear Information System (INIS)

    Chronic inflammation is characterized by the influx of neutrophils and is associated with an increased production of reactive oxygen species that can damage DNA. Oxidative DNA damage is generally thought to be involved in the increased risk of cancer in inflamed tissues. We previously demonstrated that activated neutrophil mediated oxidative stress results in a reduction in nucleotide excision repair (NER) capacity, which could further enhance mutagenesis. Inflammation and oxidative stress are critical factors in the progression of nonalcoholic fatty liver disease that is linked with enhanced liver cancer risk. In this report, we therefore evaluated the role of neutrophils and the associated oxidative stress in damage recognition and DNA repair in steatotic livers of 35 severely obese subjects with either nonalcoholic steatohepatitis (NASH) (n = 17) or steatosis alone (n = 18). The neutrophilic influx in liver was assessed by myeloperoxidase (MPO) staining and the amount of oxidative DNA damage by measuring M1dG adducts. No differences in M1dG adduct levels were observed between patients with or without NASH and also not between individuals with high or low MPO immunoreactivity. However, we found that high expression of MPO in the liver, irrespective of disease status, reduced the damage recognition capacity as determined by staining for histone 2AX phosphorylation (γH2AX). This reduction in γH2AX formation in individuals with high MPO immunoreactivity was paralleled by a significant decrease in NER capacity as assessed by a functional repair assay, and was not related to cell proliferation. Thus, the observed reduction in NER capacity upon hepatic inflammation is associated with and may be a consequence of reduced damage recognition. These findings suggest a novel mechanism of liver cancer development in patients with nonalcoholic fatty liver disease.

  13. Decreased nucleotide excision repair in steatotic livers associates with myeloperoxidase-immunoreactivity

    Energy Technology Data Exchange (ETDEWEB)

    Schults, Marten A.; Nagle, Peter W. [Department of Toxicology, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Rensen, Sander S. [Department of Surgery, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Godschalk, Roger W. [Department of Toxicology, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Munnia, Armelle; Peluso, Marco [Cancer Risk Factor Branch, ISPO Cancer Prevention and Research Institute, Via Cosimo il Vecchio 2, 50139 Florence (Italy); Claessen, Sandra M. [Department of Toxicogenomics, GROW-School for Oncology and Developmental Biology, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Greve, Jan W. [Department of Surgery, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Driessen, Ann [Department of Pathology, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Verdam, Froukje J.; Buurman, Wim A. [Department of Surgery, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Schooten, Frederik J. van [Department of Toxicology, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Chiu, Roland K., E-mail: [Department of Toxicology, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands)


    Chronic inflammation is characterized by the influx of neutrophils and is associated with an increased production of reactive oxygen species that can damage DNA. Oxidative DNA damage is generally thought to be involved in the increased risk of cancer in inflamed tissues. We previously demonstrated that activated neutrophil mediated oxidative stress results in a reduction in nucleotide excision repair (NER) capacity, which could further enhance mutagenesis. Inflammation and oxidative stress are critical factors in the progression of nonalcoholic fatty liver disease that is linked with enhanced liver cancer risk. In this report, we therefore evaluated the role of neutrophils and the associated oxidative stress in damage recognition and DNA repair in steatotic livers of 35 severely obese subjects with either nonalcoholic steatohepatitis (NASH) (n = 17) or steatosis alone (n = 18). The neutrophilic influx in liver was assessed by myeloperoxidase (MPO) staining and the amount of oxidative DNA damage by measuring M{sub 1}dG adducts. No differences in M{sub 1}dG adduct levels were observed between patients with or without NASH and also not between individuals with high or low MPO immunoreactivity. However, we found that high expression of MPO in the liver, irrespective of disease status, reduced the damage recognition capacity as determined by staining for histone 2AX phosphorylation ({gamma}H2AX). This reduction in {gamma}H2AX formation in individuals with high MPO immunoreactivity was paralleled by a significant decrease in NER capacity as assessed by a functional repair assay, and was not related to cell proliferation. Thus, the observed reduction in NER capacity upon hepatic inflammation is associated with and may be a consequence of reduced damage recognition. These findings suggest a novel mechanism of liver cancer development in patients with nonalcoholic fatty liver disease.

  14. Verification of the Cross Immunoreactivity of A60, a Mouse Monoclonal Antibody against Neuronal Nuclear Protein (United States)

    Mao, Shanping; Xiong, Guoxiang; Zhang, Lei; Dong, Huimin; Liu, Baohui; Cohen, Noam A.; Cohen, Akiva S.


    A60, the mouse monoclonal antibody against the neuronal nuclear protein (NeuN), is the most widely used neuronal marker in neuroscience research and neuropathological assays. Previous studies identified fragments of A60-immunoprecipitated protein as Synapsin I (Syn I), suggesting the antibody will demonstrate cross immunoreactivity. However, the likelihood of cross reactivity has never been verified by immunohistochemical techniques. Using our established tissue processing and immunofluorescent staining protocols, we found that A60 consistently labeled mossy fiber terminals in hippocampal area CA3. These A60-positive mossy fiber terminals could also be labeled by Syn I antibody. After treating brain slices with saponin in order to better preserve various membrane and/or vesicular proteins for immunostaining, we observed that A60 could also label additional synapses in various brain areas. Therefore, we used A60 together with a rabbit monoclonal NeuN antibody to confirm the existence of this cross reactivity. We showed that the putative band positive for A60 and Syn I could not be detected by the rabbit anti-NeuN in Western blotting. As efficient as Millipore A60 to recognize neuronal nuclei, the rabbit NeuN antibody demonstrated no labeling of synaptic structures in immunofluorescent staining. The present study successfully verified the cross reactivity present in immunohistochemistry, cautioning that A60 may not be the ideal biomarker to verify neuronal identity due to its cross immunoreactivity. In contrast, the rabbit monoclonal NeuN antibody used in this study may be a better candidate to substitute for A60. PMID:27242450

  15. Comparative mapping of GABA-immunoreactive neurons in the central nervous systems of nudibranch molluscs. (United States)

    Gunaratne, Charuni A; Sakurai, Akira; Katz, Paul S


    The relative simplicity of certain invertebrate nervous systems, such as those of gastropod molluscs, allows behaviors to be dissected at the level of small neural circuits composed of individually identifiable neurons. Elucidating the neurotransmitter phenotype of neurons in neural circuits is important for understanding how those neural circuits function. In this study, we examined the distribution of γ-aminobutyric-acid;-immunoreactive (GABA-ir) neurons in four species of sea slugs (Mollusca, Gastropoda, Opisthobranchia, Nudibranchia): Tritonia diomedea, Melibe leonina, Dendronotus iris, and Hermissenda crassicornis. We found consistent patterns of GABA immunoreactivity in the pedal and cerebral-pleural ganglia across species. In particular, there were bilateral clusters in the lateral and medial regions of the dorsal surface of the cerebral ganglia as well as a cluster on the ventral surface of the pedal ganglia. There were also individual GABA-ir neurons that were recognizable across species. The invariant presence of these individual neurons and clusters suggests that they are homologous, although there were interspecies differences in the numbers of neurons in the clusters. The GABAergic system was largely restricted to the central nervous system, with the majority of axons confined to ganglionic connectives and commissures, suggesting a central, integrative role for GABA. GABA was a candidate inhibitory neurotransmitter for neurons in central pattern generator (CPG) circuits underlying swimming behaviors in these species, however none of the known swim CPG neurons were GABA-ir. Although the functions of these GABA-ir neurons are not known, it is clear that their presence has been strongly conserved across nudibranchs. PMID:24638845

  16. Sodium channel Nav1.7 immunoreactivity in painful human dental pulp and burning mouth syndrome

    Directory of Open Access Journals (Sweden)

    Yiangou Yiangos


    Full Text Available Abstract Background Voltage gated sodium channels Nav1.7 are involved in nociceptor nerve action potentials and are known to affect pain sensitivity in clinical genetic disorders. Aims and Objectives To study Nav1.7 levels in dental pulpitis pain, an inflammatory condition, and burning mouth syndrome (BMS, considered a neuropathic orofacial pain disorder. Methods Two groups of patients were recruited for this study. One group consisted of patients with dental pulpitis pain (n = 5 and controls (n = 12, and the other patients with BMS (n = 7 and controls (n = 10. BMS patients were diagnosed according to the International Association for the Study of Pain criteria; a pain history was collected, including the visual analogue scale (VAS. Immunohistochemistry with visual intensity and computer image analysis were used to evaluate levels of Nav1.7 in dental pulp tissue samples from the dental pulpitis group, and tongue biopsies from the BMS group. Results There was a significantly increased visual intensity score for Nav1.7 in nerve fibres in the painful dental pulp specimens, compared to controls. Image analysis showed a trend for an increase of the Nav1.7 immunoreactive % area in the painful pulp group, but this was not statistically significant. When expressed as a ratio of the neurofilament % area, there was a strong trend for an increase of Nav1.7 in the painful pulp group. Nav1.7 immunoreactive fibres were seen in abundance in the sub-mucosal layer of tongue biopsies, with no significant difference between BMS and controls. Conclusion Nav1.7 sodium channel may play a significant role in inflammatory dental pain. Clinical trials with selective Nav1.7 channel blockers should prioritise dental pulp pain rather than BMS.

  17. The transcriptional landscape

    DEFF Research Database (Denmark)

    Nielsen, Henrik


    The application of new and less biased methods to study the transcriptional output from genomes, such as tiling arrays and deep sequencing, has revealed that most of the genome is transcribed and that there is substantial overlap of transcripts derived from the two strands of DNA. In protein coding...... independent transcription from within the unit. In genomic regions separating those that encode proteins or highly abundant RNA molecules with known function, transcripts are generally of low abundance and short-lived. In most of these cases, it is unclear to what extent a function is related to transcription...

  18. Histamine Immunoreactive Elements in the Central and Peripheral Nervous Systems of the Snail, Biomphalaria spp., Intermediate Host for Schistosoma mansoni.

    Directory of Open Access Journals (Sweden)

    Mohamed R Habib

    Full Text Available Histamine appears to be an important transmitter throughout the Animal Kingdom. Gastropods, in particular, have been used in numerous studies establishing potential roles for this biogenic amine in the nervous system and showing its involvement in the generation of diverse behaviours. And yet, the distribution of histamine has only previously been described in a small number of molluscan species. The present study examined the localization of histamine-like immunoreactivity in the central and peripheral nervous systems of pulmonate snails of the genus Biomphalaria. This investigation demonstrates immunoreactive cells throughout the buccal, cerebral, pedal, left parietal and visceral ganglia, indicative of diverse regulatory functions in Biomphalaria. Immunoreactivity was also present in statocyst hair cells, supporting a role for histamine in graviception. In the periphery, dense innervation by immunoreactive fibers was observed in the anterior foot, perioral zone, and other regions of the body wall. This study thus shows that histamine is an abundant transmitter in these snails and its distribution suggest involvement in numerous neural circuits. In addition to providing novel subjects for comparative studies of histaminegic neurons in gastropods, Biomphalaria is also the major intermediate host for the digenetic trematode parasite, which causes human schistosomiasis. The study therefore provides a foundation for understanding potential roles for histamine in interactions between the snail hosts and their trematode parasites.

  19. Differential Immuno-Reactivity to Genomic DNA, RNA and Mitochondrial DNA is Associated with Auto-Immunity

    Directory of Open Access Journals (Sweden)

    Vilena V. Ivanova


    Full Text Available Background: Circulating auto-reactive antibodies are hallmark features of auto-immune diseases, however little is known with respect to the specificity of such bio-markers. In the present study, we investigated the specificity of anti-nucleic acid antibodies in the blood of subjects with systemic lupus erythematosus (SLE and healthy controls. Methods: Sera from 12 SLE cases and 8 controls were evaluated for immuno-reactivity to purified RNA, DNA and mitochondrial DNA (mtDNA by enzyme-linked immuno-sorbent assay (ELISA. Results: As expected, immuno-reactivity to total nucleic acids was significantly higher in subjects with SLE when compared to healthy controls, however a clear distinction was observed among the various nucleic acid sub-types, with sera from SLE subjects displaying the greatest immuno-reactivity to RNA followed by mtDNA and then total DNA. Conclusion: The identification of auto-reactive antibodies can serve as highly sensitive biomarkers, although their specificity may not always allow diagnostic certainty. The knowledge that auto-antibodies in subjects with SLE display differential immuno-reactivity may help to improve existing diagnostics and may lead to a better understanding of the pathogenesis of auto-immune disorders.

  20. Characterization of β-endorphin-immunoreactivity in limbic brain structures of rats self-administering heroin or cocaine

    NARCIS (Netherlands)

    Sweep, C.G.J.; Ree, J.M. van; Wiegant, V.M.


    The effects of intravenous self-administration of 30 μg infusions of either heroin or cocaine, or saline on the concentrations of β-endorphin-immunoreactivity (βE-IR) in the anterior part of the rat brain limbic system were studied. Self-administration of heroin and cocaine for 5 daily sessions resu

  1. Substance P immunoreactivity in the lumbar spinal cord of the turtle Trachemys dorbigni following peripheral nerve injury

    Directory of Open Access Journals (Sweden)

    W.A. Partata


    Full Text Available Immunoreactive substance P was investigated in turtle lumbar spinal cord after sciatic nerve transection. In control animals immunoreactive fibers were densest in synaptic field Ia, where the longest axons invaded synaptic field III. Positive neuronal bodies were identified in the lateral column of the dorsal horn and substance P immunoreactive varicosities were observed in the ventral horn, in close relationship with presumed motoneurons. Other varicosities appeared in the lateral and anterior funiculi. After axotomy, substance P immunoreactive fibers were reduced slightly on the side of the lesion, which was located in long fibers that invaded synaptic field III and in the varicosities of the lateral and anterior funiculus. The changes were observed at 7 days after axonal injury and persisted at 15, 30, 60 and 90 days after the lesion. These findings show that turtles should be considered as a model to study the role of substance P in peripheral axonal injury, since the distribution and temporal changes of substance P were similar to those found in mammals.

  2. Plant transcription factors. (United States)

    Meshi, T; Iwabuchi, M


    Transcriptional regulation of gene expression relies on the recognition of promoter elements by transcription factors. In the past several years, a considerable number of (putative) transcription factors have been identified in plants. Some genes coding for these factors were isolated by south-western screening with oligonucleotides as a probe or by homology-based screening, and others were initially isolated by genetic means and subsequently identified as the genes for transcription factors. These transcription factors often form families of structurally related proteins with similar DNA-binding specificities and in addition, they are sometimes involved in related phenomena. Some groups of factors homo- and/or heterodimerize to increase the length and variability of the target sequences. Transcriptional activators, in general, comprise a modular activation domain. The activities of the transcription factors are controlled by post-translational modification, like phosphorylation and glycosylation, as well as at the levels of nuclear transport, oligomerization, etc. In this review, we will summarize the current knowledge of plant transcription factors to help understand the mechanistic aspects of the transcriptional regulation of genes. PMID:8589926

  3. Somatostatin immunoreactive cells in lesional psoriatic human skin during peptide T treatment. (United States)

    Johansson, O; Hilliges, M; Talme, T; Marcusson, J A; Wetterberg, L


    Peptide T has been shown to be an effective treatment in psoriasis. The mechanism through which peptide T works in psoriasis is at present unknown. Furthermore, a clearance of psoriasis has also been registered using the inhibitory peptide somatostatin. These observations all focus on the fact that peptide T, somatostatin, and/or other peptides, might provide a clue to understanding the etiology and pathogenesis of psoriasis. Therefore, the effect of peptide T administration on somatostatin containing cutaneous cell populations was investigated. Ten psoriatic patients were treated with peptide T (D-Ala-peptide T amide; 2 mg/day i.v.) for 28 days. Serial biopsies were obtained from the psoriatic lesions before, once weekly during and 4 weeks after discontinuation of the peptide T treatment. An indirect immunofluorescence procedure was performed using a polyclonal antiserum against somatostatin. Clinically, most of the patients responded successfully to the treatment. Immunohistochemical investigations of the serial biopsies revealed the appearance of extensive changes in the number of dermal somatostatin immunoreactive dendritic cells. We believe that peptide T may stimulate the local synthesis and/or release of somatostatin, or proliferation and/or migration of certain dendritic cell populations in psoriatic lesions during healing. Since the benefits of peptide T treatment of psoriatic patients parallel earlier investigations using somatostatin infusions, it is likely that somatostatin given exogenously or synthesized/released endogenously plays a vital role in inducing the healing process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7911613

  4. Rearrangement of S-100 immunoreactive Langerhans' cells in human psoriatic skin treated with peptide T. (United States)

    Wang, L; Hilliges, M; Talme, T; Marcusson, J A; Wetterberg, L; Johansson, O


    Dendritic cells marked by protein S-100 (S-100) antiserum in the suprabasal layers of the epidermis have previously been identified to be Langerhans' cells. In this study, S-100 immunoreactive cells have been investigated in psoriatic lesioned skin during and after peptide T treatment. Peptide T is an octapeptide with affinity for the CD4 receptor. Nine patients were intravenously infused with peptide T, 2 mg in 500 ml saline per day for 28 days. Sections from involved skin before, every week during, and after the treatment were processed by indirect immunofluorescence using S-100 antiserum. Before the treatment the epidermal Langerhans' cells were numerically decreased or even completely gone in the involved skin of psoriasis as compared to skin from normal healthy controls, while the dermal dendritic cells instead were increased and gathered in cell clusters around vascular structures. Four of the nine patients had histopathological improvements after the peptide T treatment, and, in those cases, the dendritic cells in the dermis were reduced in number, and the Langerhans' cells in the epidermis were numerically increased as well as even reversed to normal position and morphology. These changes in the distribution and density of Langerhans' cells represent their rearrangement during the course of psoriasis and/or the remission after peptide T treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7727353

  5. Distribution of hypothalamic vasoactive intestinal peptide immunoreactive neurons in the male native Thai chicken. (United States)

    Kamkrathok, Boonyarit; Sartsoongnoen, Natagarn; Prakobsaeng, Nattiya; Rozenboim, Israel; Porter, Tom E; Chaiseha, Yupaporn


    Avian prolactin (PRL) secretion is under stimulatory control by the PRL-releasing factor (PRF), vasoactive intestinal peptide (VIP). The neuroendocrine regulation of the avian reproductive system has been extensively studied in females. However, there are limited data in males. The aim of this study was to elucidate the VIPergic system and its relationship to PRL and testosterone (T) in the male native Thai chicken. The distributions of VIP-immunoreactive (-ir) neurons and fibers were determined by immunohistochemistry. Changes in VIP-ir neurons within the nucleus inferioris hypothalami (IH) and nucleus infundibuli hypothalami (IN) areas were compared across the reproductive stages. Plasma levels of PRL and T were determined by enzyme-linked immunosorbent assay and then compared across the reproductive stages. The results revealed that the highest accumulations of VIP-ir neurons were concentrated only within the IH-IN, and VIP-ir neurons were not detected within other hypothalamic nuclei. Within the IH-IN, VIP-ir neurons were low in premature and aging males and markedly increased in mature males. Changes in VIP-ir neurons within the IH-IN were directly mirrored with changes in PRL and T levels across the reproductive stages. These results suggested that VIP neurons in the IH-IN play a regulatory role in year-round reproductive activity in males. The present study also provides additional evidence that VIP is the PRF in non-seasonal, continuously breeding equatorial species. PMID:27269881

  6. Alzheimer's-associated Aβ oligomers show altered structure, immunoreactivity and synaptotoxicity with low doses of oleocanthal

    International Nuclear Information System (INIS)

    It now appears likely that soluble oligomers of amyloid-β1-42 peptide, rather than insoluble fibrils, act as the primary neurotoxin in Alzheimer's disease (AD). Consequently, compounds capable of altering the assembly state of these oligomers (referred to as ADDLs) may have potential for AD therapeutics. Phenolic compounds are of particular interest for their ability to disrupt Aβ oligomerization and reduce pathogenicity. This study has focused on oleocanthal (OC), a naturally-occurring phenolic compound found in extra-virgin olive oil. OC increased the immunoreactivity of soluble Aβ species, when assayed with both sequence- and conformation-specific Aβ antibodies, indicating changes in oligomer structure. Analysis of oligomers in the presence of OC showed an upward shift in MW and a ladder-like distribution of SDS-stable ADDL subspecies. In comparison with control ADDLs, oligomers formed in the presence of OC (Aβ-OC) showed equivalent colocalization at synapses but exhibited greater immunofluorescence as a result of increased antibody recognition. The enhanced signal at synapses was not due to increased synaptic binding, as direct detection of fluorescently-labeled ADDLs showed an overall reduction in ADDL signal in the presence of OC. Decreased binding to synapses was accompanied by significantly less synaptic deterioration assayed by drebrin loss. Additionally, treatment with OC improved antibody clearance of ADDLs. These results indicate oleocanthal is capable of altering the oligomerization state of ADDLs while protecting neurons from the synaptopathological effects of ADDLs and suggest OC as a lead compound for development in AD therapeutics.

  7. Proteomic analysis of Ascaridia galli. Identification of immunoreactive proteins in naturally and experimentally infected hens. (United States)

    González-Miguel, Javier; Marcos-Atxutegi, Cristina; de Castello, Roberta Bottari; Carpani, Sara; Morchón, Rodrigo; Simón, Fernando


    Ascaridia galli, intestinal parasite of domestic fowl, is responsible of economic losses in avian exploitations. However, molecular mechanisms that govern avian ascaridiasis remain largely unknown. The aim of the present work was to identify proteins of A. galli recognized by the immune system of naturally and experimentally infected hens, using two-dimensional electrophoresis (2-DE) and mass spectrometry (MS). Sixteen immunoreactive proteins of A. galli were identified. These proteins are mainly related to different metabolic processes, cell motility and binding activities. The timing evolution of this recognition pattern was studied using serum samples from experimentally infected hens, allowing us to observe an early recognition of many of these antigens. Many of them were isoforms from lipid and plasminogen-binding proteins. Moreover, plasminogen-binding activity has been related in other parasites with the facilitation of intra-organic migration, which represents an important fact in avian ascaridiasis. This work represents the first proteomic study of A. galli and could contribute to explain some aspects of parasite/host relationships of avian ascaridiasis. PMID:23578998

  8. Rapid lymphocyte immunoreactivity test utilizing [3H]uridine in vitro

    International Nuclear Information System (INIS)

    A microculture assay utilizing [3H]uridine incorporation was developed to test murine spleen lymphocyte immunoreactivity in vitro. Parameters of the culture technique which included cell density, doses of LPS, Con A, PHA, [3H]uridine levels, and length of culture time were investigated. Responses were detectable at 4 h for all 3 mitogens, with labelling ranging up to 180% of the control value. By 8 h there was a 200-350% increase in mitogen-induced incorporation of radioactivity. Similar increases were observed in a serum-free system. The responses were the result of increased incorporation of label by stimulated cultures rather than decreased labeling of non-mitogen treated cultures over time. The [3H]uridine incorporation was demonstrated to be the selective response of T or B cell populations when stimulated with appropriate lectins. This assay detects early RNA synthesis, as supported by experimental observations in which accumulation of radioactivity in stimulated lymphocytes was TCA precipitable, resistant to SDS treatment, and inhibited by actinomycin D. (Auth.)

  9. Predictive value of bcl-2 immunoreactivity in prostate cancer patients treated with radiotherapy

    International Nuclear Information System (INIS)

    Background and purpose: Recent experimental evidence suggests that overexpression of bcl-2, a protein functioning by blocking apoptosis, may influence the treatment outcome in human tumours, including prostate cancer. To test the clinical implications of this hypothesis, tumours from patients with prostate cancer treated with external beam radiotherapy were investigated for bcl-2 immunoreactivity (IR) and correlated with prognosis and treatment outcome. Materials and methods: Bcl-2 IR was evaluated in archival tumour specimens obtained through transurethral resection from 42 patients with localized prostate cancer (T0-T4, N0 and M0). Bcl-2 IR expression was related to stage, grade and cancer-specific survival. Specimens were obtained prior to administrating routine radiotherapy for all patients. Results: Bcl-2 IR was present in 19/42 (45%) tumours. The bcl-2-positive patients had a significantly longer cancer-specific survival than the bcl-2-negative patients (10.3 versus 3.4 years, P<0.04). At follow-up (7-19 years), nine patients were still alive, 26 patients had died of prostate cancer and seven patients had died of other causes. Conclusions: This study indicates that pre-treatment bcl-2 overexpression is related to a favourable outcome in prostate cancer treated with radiotherapy. Low bcl-2 along with a high stage may be a predictor of poor prognosis and these patients might benefit from additional treatment. (Copyright (c) 1998 Elsevier Science B.V., Amsterdam. All rights reserved.)

  10. Localization of GABA-like immunoreactivity in the central nervous system of Aplysia californica. (United States)

    Díaz-Ríos, M; Suess, E; Miller, M W


    Gamma-aminobutyric acid (GABA) is present in the central nervous system of Aplysia californica (Gastropoda, Opisthobranchia) where its role as a neurotransmitter is supported by pharmacological, biochemical, and anatomical investigations. In this study, the distribution of GABA-immunoreactive (GABAi) neurons and fiber systems in Aplysia was examined by using wholemount immunohistochemistry and nerve backfill methods. GABAi neurons were located in the buccal, cerebral, and pedal ganglia. Major commissural fiber systems were present in each of these ganglia, whereas more limited fiber systems were observed in the ganglionic connectives. Some of the interganglionic fibers were found to originate from two unpaired GABAi neurons, one in the buccal ganglion and one in the right pedal ganglion, each of which exhibited bilateral projections. No GABAi fibers were found in the nerves that innervate peripheral sensory, motor, or visceral organs. Although GABAi cells were not observed in the pleural or abdominal ganglia, these ganglia did receive limited projections of GABAi fibers originating from neurons in the pedal ganglia. The distribution of GABAi neurons suggests that this transmitter system may be primarily involved in coordinating certain bilateral central pattern generator (CPG) systems related to feeding and locomotion. In addition, the presence of specific interganglionic GABAi projections also suggests a role in the regulation or coordination of circuits that produce components of complex behaviors. PMID:10524338

  11. Serotonin immunoreactivity in the central nervous system of the marine molluscs Pleurobranchaea californica and Tritonia diomedea. (United States)

    Sudlow, L C; Jing, J; Moroz, L L; Gillette, R


    The central nervous systems of the marine molluscs Pleurobranchaea californica (Opisthobranchia: Notaspidea) and Tritonia diomedea (Opisthobranchia: Nudibranchia) were examined for serotonin-immunoreactive (5-HT-IR) neurons and processes. Bilaterally paired clusters of 5-HT-IR neuron somata were distributed similarly in ganglia of the two species. In the cerebropleural ganglion complex, these were the metacerebral giant neurons (both species), a dorsal anterior cluster (Pleurobranchaea only), a dorsal medial cluster including identified neurons of the escape swimming network (both species), and a dorsal lateral cluster in the cerebropleural ganglion (Pleurobranchaea only). A ventral anterior cluster (both species) adjoined the metacerebral giant somata at the anterior ganglion edge. Pedal ganglia had the greatest number of 5-HT-IR somata, the majority located near the roots of the pedal commissure in both species. Most 5-HT-IR neurons were on the dorsal surface of the pedal ganglia in Pleurobranchaea and were ventral in Tritonia. Neither the buccal ganglion of both species nor the visceral ganglion of Pleurobranchaea had 5-HT-IR somata. Afew asymmetrical 5-HT-IR somata were found in cerebropleural and pedal ganglia in both species, always on the left side. The clustering of 5-HT-IR neurons, their diverse axon pathways, and the known physiologic properties of their identified members are consistent with a loosely organized arousal system of serotonergic neurons whose components can be generally or differentially active in expression of diverse behaviors. PMID:9619500

  12. Immunoreactivity of S100β protein in the hippocampus of chinchilla

    Directory of Open Access Journals (Sweden)

    Krawczyk Aleksandra


    Full Text Available The aim of the study was to investigate S100β protein in astrocytes of CA1 and CA3 areas of the hippocampus proper and the dentate gyrus with the hilus yet undefined in mature males of chinchilla. The presence of S100β was determined using indirect immunohistochemical peroxidase-antiperoxidase method with specific monoclonal antibody against this protein. Most of the S100β-positive cells were detected in the subgranular zone of the dentate gyrus and in the middle part of the hilus. In CA3 area, it was found that the most numerous cells with S100β are in stratum radiatum. In CA1 area, there were single astrocytes expressing this protein. This data demonstrates species differences and a large quantity of S100β immunoreactive cells in the subgranular zone of the dentate gyrus of chinchilla, which may be associated with structural reorganisation of the hippocampus and with neurogenesis, learning, and memorising process dependent on the hippocampus.

  13. Effects of Shenpang acupoint-stimulation on estrogen receptor immunoreactive neurons in thalamus of rabbits

    Institute of Scientific and Technical Information of China (English)

    LUO Qihui; CHEN Zhengli; ZHU Chunmei; FAN Guangli; HUANG Yidan


    To investigate the effects of Shenpang acupoint-stimulation in reproductive endocrinology,the changes in estrogen receptor immunoreactive (ER-IR)neurons after Shenpang acupoint-stimulation were studied by using immnunohistochemistry.ER-IR positive reactions were detected in most nuclei of the thalamus.In the acupuncturetreated group,a great number of ER-IR positive neurons with clear dendrites existed in the nucleus,paraventricular nucleus,ventrolateral nucleus,ventromedial nucleus,ventroprincipal nucleus,centromedian nucleus,reticular nucleus,and periventricular nucleus of thalamus,and they were strongly stained.In addition,the ER-IR positive neurons were mainly located in the cytoplasm,nucleus and neutrite,and some also existed in the cytoplasmic membrane.In contrast,a few neurons existed in the above-mentioned nuclei in the control group,but they were slightly stained.It is concluded that Shenpang acupoint-stimulation can promote the expression of estrogen receptors in the above nuclei.

  14. Antigen-binding site protection during radiolabeling leads to a higher immunoreactive fraction

    International Nuclear Information System (INIS)

    It is generally accepted that the immunointegrity of an antibody (Ab) depends on the preservation of its antigen-binding sites. Our goal was to radiolabel an antibody at several iodine:antibody molar ratios under conditions protecting its combining site and to compare its immunoreactive fraction (IRF) and electrophoretic mobility with those of the same antibody radiolabeled without protection. The data indicate that an antibody radiolabeled while its antigen-binding site is occupied by its antigen had the same IRF, regardless of the number of iodine atoms per antibody molecule. On the other hand, even at an I:Ab ratio of 1:1, the IRF of the same antibody radiolabeled without protection was lower than that of a protected one and decreased with increasing I:Ab ratios. In addition, the iodination of these Ab changes their electrophoretic mobility; however, when the Ab is labeled in the protected state, the degree of change is less. The binding of an antibody to its antigen prior to radiolabeling, therefore, enhances its immuno-integrity and prevents major conformational changes as reflected by electrophoresis

  15. Neuroanatomy of pars intercerebralis neurons with special reference to their connections with neurons immunoreactive for pigment-dispersing factor in the blow fly Protophormia terraenovae. (United States)

    Yasuyama, Kouji; Hase, Hiroaki; Shiga, Sakiko


    Input regions of pars intercerebralis (PI) neurons are examined by confocal and electron microscopies with special reference to their connections with neurons immunoreactive for pigment-dispersing factor (PDF) in the blow fly, Protophormia terraenovae. PI neurons are a prerequisite for ovarian development under long-day conditions. Backfills from the cardiac recurrent nerve after severance of the posterior lateral tracts labeled thin fibers derived from the PI neurons in the superior medial protocerebrum. These PI fibers were mainly synapsin-negative and postsynaptic to unknown varicose profiles containing dense-core vesicles. Backfilled fibers in the periesophageal neuropils, derived from the PI neurons or neurons with somata in the subesophageal zone, were varicose and some were synapsin-positive. Electron microscopy revealed the presence of both presynaptic and postsynaptic sites in backfilled fibers in the periesophageal neuropils. Many PDF-immunoreactive varicosities were found in the superior medial and lateral protocerebrum and double-labeling showed that 60-88 % of PDF-immunoreactive varicosities were also synapsin-immunoreactive. Double-labeling with the backfills and PDF immunocytochemistry showed that the PI fibers and PDF-immunoreactive varicosities were located close to each other in the superior medial protocerebrum. Results of triple-labeling of PI neurons, PDF-immunoreactive neurons and synapsin-immunoreactive terminals demonstrated that the synapsin-positive PDF-immunoreactive varicosities contacted the PI fibers. These data suggest that PI neurons receive synaptic contacts from PDF-immunoreactive fibers, which are derived from circadian clock neurons, of small ventral lateral neurons (previously called OL2) or posterior dorsal (PD) neurons with somata in the pars lateralis. PMID:25971932

  16. The Transcription Factor Encyclopedia

    DEFF Research Database (Denmark)

    Yusuf, Dimas; Butland, Stefanie L; Swanson, Magdalena I;


    ABSTRACT: Here we present the Transcription Factor Encyclopedia (TFe), a new web-based compendium of mini review articles on transcription factors (TFs) that is founded on the principles of open access and collaboration. Our consortium of over 100 researchers has collectively contributed over 130...

  17. Some enkephalin- or VIP-immunoreactive hippocampal pyramidal cells contain neurofibrillary tangles in the brains of aged humans and persons with Alzheimer's disease. (United States)

    Kulmala, H K


    Neurofibrillary tangles are one of the histopathological neuronal abnormalities present in normal aging and especially in Alzheimer's Disease. We have utilized immunocytochemical staining for neuropeptides followed by Congo red with gallocyanin counterstaining and polarized illumination to determine whether enkephalin (Enk), somatostatin (Som), cholecystokinin (CCK), or vasoactive intestinal polypeptide (VIP) are contained in neurons afflicted with such tangles. A few Enk- or VIP-immunoreactive pyramidal cells in field hl and subiculum were found to contain tangles. Many such Enk- or VIP-immunoreactive neurons and cells containing Som- or CCK-like immunoreactivity did not contain such tangles. PMID:2410823

  18. FMRFamide-like immunoreactivity in the central nervous system of the cephalopod mollusc, Idiosepius notoides

    DEFF Research Database (Denmark)

    Wollesen, Tim; Loesel, R; Wanninger, Andreas Wilhelm Georg


    For more than a century, cephalopod molluscs have been the subject of extensive studies with respect to their complex neuroanatomy and behavior. In comparison to gastropod molluscs surprisingly little work has been carried out on the characterization of neurons in the central nervous system (CNS......) of cephalopods with respect to their neurotransmitter phenotypes. This study presents preliminary results on the distribution of FMRFamide-like immunoreactive neurons within the CNS of the pygmy squid Idiosepius notoides . Its gross neuroanatomy resembles that of other cephalopods. FMRFamide...... the vertical lobes. Although certain immunohistochemical traits are shared with other cephalopods, such as a wall-like arrangement of FMRFamide-like immunoreactive cell somata within the dorsal basal lobe, others have so far only been found in Idiosepius . However, future investigations on other...

  19. Increase of TRPV1-Immunoreactivity in Dorsal Root Ganglia Neurons Innervating the Femur in a Rat Model of Osteoporosis


    Yoshino, Kensuke; Suzuki, Miyako; Kawarai, Yuya; Sakuma, Yoshihiro; Inoue, Gen; Orita, Sumihisa; Yamauchi, Kazuyo; Aoki, Yasuchika; Ishikawa, Tetsuhiro; Miyagi, Masayuki; Kamoda, Hiroto; Kubota, Gou; Oikawa, Yasuhiro; Inage, Kazuhide; Sainoh, Takeshi


    Purpose Transient receptor potential vanilloid 1 (TRPV1) is a ligand-gated nonselective cation channel, which can be activated by capsaicin and other noxious stimuli. Recently, an association between bone pain and TRPV1 has been reported. However, the influence of osteoporosis on TRPV1 in the sensory system innervating the femur has not been reported. Materials and Methods TRPV1-immunoreactive (ir) in dorsal root ganglia (DRG) neurons labeled with neurotracer [Fluoro-Gold (FG)] innervating th...

  20. Effects of Static Magnetic Field on Growth of Leptospire, Leptospira interrogans serovar canicola: Immunoreactivity and Cell Division


    Triampo, Wannapong; Doungchawee, Galayanee; Triampo, Darapond; Wong-Ekkabut, Jirasak; Tang, I-Ming


    The effects of the exposure of the bacterium, Leptospira interrogans serovar canicola to a constant magnetic field with magnetic flux density from a permanent ferrite magnet = 140 mT were studied. Changes in Leptospira cells after their exposure to the field were determined on the basis of changes in their growth behavior and agglutination immunoreactivity with a homologous antiserum using darkfield microscopy together with visual imaging. The data showed that the exposed Leptospira cells hav...

  1. Changes in small intestinal chromogranin A-immunoreactive cell densities in patients with irritable bowel syndrome after receiving dietary guidance


    Mazzawi, Tarek; El-Salhy, Magdy


    Chromogranin A (CgA) is a common marker for enteroendocrine cells in the gut, and CgA-immunoreactive cell densities are abnormal in patients with irritable bowel syndrome (IBS). The majority of patients with IBS report that their symptoms develop after consuming certain foodstuffs. In the present study, we investigated the effects of dietary guidance on the total enteroendocrine cell densities in the small intestine, as detected by CgA. A total of 14 patients with IBS underwent a gastroscopy ...

  2. Lack of relationship between TIMP-1 tumour cell immunoreactivity, treatment efficacy and prognosis in patients with advanced epithelial ovarian cancer

    International Nuclear Information System (INIS)

    Tissue inhibitor of metalloproteinase 1 (TIMP-1) is a natural inhibitor of the matrix metalloproteinases (MMPs) which are proteolytic enzymes involved in degradation of extracellular matrix thereby favoring tumour cell invasion and metastasis. TIMP-1 activity in tumour tissue may therefore play an essential role in the progression of a malignant tumour. The primary aim of the present study was to evaluate TIMP-1 protein immunoreactivity in tissue from primary ovarian cancer patients and associate these findings with the course of the disease including response to treatment in the individual patient. TIMP-1 was assessed by immunohistochemistry (in tissue micro arrays) in a total of 163 ovarian cancer specimens obtained from primary debulking surgery during 1991-1994 as part of a randomized clinical protocol. Positive TIMP-1 immunoreactivity was found in 12.3% of the tumours. The median survival time for the 143 patients with TIMP-1 negative tumours was 23.7 months [19.0-29.4] 95% CI, while the median survival time for the 20 patients with TIMP-1 positive tumours was 15.9 months [12.3-27.4] 95% CI. Although a difference of 7.8 months in median overall survival in favor of the TIMP-1 tumour negative patients was found, this difference did not reach statistical significance (p = 0.28, Kaplan-Meier, log-rank test). Moreover, TIMP-1 immunoreactivity was not associated with CA125 response (p = 0.53) or response at second look surgery (p = 0.72). TIMP-1 immunoreactivity in tumour tissue from patients with primary epithelial ovarian cancer did not correlate with patient survival or response to combination platinum/cyclophosphamide therapy

  3. Reelin-immunoreactive neurons in entorhinal cortex layer II selectively express intracellular amyloid in early Alzheimer's disease. (United States)

    Kobro-Flatmoen, Asgeir; Nagelhus, Anne; Witter, Menno P


    The onset of Alzheimer's disease (AD) is associated with subtle pathological changes including increased intracellular expression of amyloid-β (Aβ). A structure affected particularly early in the course of AD is the entorhinal cortex, where neuronal death in layer II is observed already at initial stages. Neurons in EC-layer II, particularly those that express the protein Reelin, give rise to projections to the hippocampal dentate gyrus and this projection shows severe loss of synaptic contacts during early-stage AD. Given this anatomical specificity, we sought to determine whether increased intracellular expression of Aβ is selectively associated with Reelin-immunoreactive neurons in layer II of the entorhinal cortex. Here we report that in a transgenic rat model, which mimics the onset and distribution of extracellular amyloid deposits seen in human AD subjects, expression of intracellular Aβ in entorhinal layer II selectively occurs in Reelin-immunoreactive neurons during the early, pre-plaque stage. This Reelin-Aβ association is also present in human subjects with AD-related pathological changes, even in early disease stages. These findings strongly indicate that Reelin-immunoreactive neurons in entorhinal layer II play a crucial role during the initial stages of AD, and may therefore lead to refined hypotheses concerning the origin of this devastating condition. PMID:27195475

  4. Calcitonin gene-related peptide immunoreactive sensory neurons in the vagal and glossopharyngeal ganglia innervating the larynx of the rat. (United States)

    Hayakawa, Tetsu; Kuwahara-Otani, Sachi; Maeda, Seishi; Tanaka, Koichi; Seki, Makoto


    We have examined whether calcitonin gene-related peptide-immunoreactive (CGRP-ir) neurons in the vagal and glossopharyngeal ganglia innervate the larynx. Many CGRP-ir neurons were located mostly in the superior glossopharyngeal-jugular ganglion complex that was fused the superior glossopharyngeal ganglion and the jugular ganglion in the cranial cavity. When Fluorogold was applied to the cut end of the superior laryngeal nerve (SLN) or the recurrent laryngeal nerve (RLN), many Fluorogold-labeled neurons were found in the superior glossopharyngeal-jugular ganglion complex and the nodose ganglion. Double-labeling for CGRP and Fluorogold showed that about 80% of Fluorogold-labeled neurons in the superior glossopharyngeal-jugular ganglion complex expressed CGRP-like immunoreactivity in the case of application to the SLN, and about 50% of Fluorogold-labeled neurons expressed CGRP-like immunoreactivity in the case of the RLN. Only a few double-labeled neurons were found in the nodose ganglion. The number of the Fluorogold-labeled neurons and double-labeled neurons in the superior glossopharyngeal-jugular ganglion complex in the case of the SLN was larger than that in the case of the RLN. These results indicate that sensory information from the larynx might be conveyed by many CGRP-ir neurons located in the superior glossopharyngeal-jugular ganglion complex by way of the SLN and the RLN. PMID:24269509

  5. Comparison of three tissue fixatives on the immunoreactivity of mammalian P-glycoprotein antibodies to teleost tissues

    Energy Technology Data Exchange (ETDEWEB)

    Hemmer, M.J. [Environmental Protection Agency, Gulf Breeze, FL (United States)]|[Univ. of Mississippi, University, MS (United States). Dept. of Pharmacology; Courtney, L.A. [Environmental Protection Agency, Gulf Breeze, FL (United States); Benson, W.H. [Univ. of Mississippi, University, MS (United States)


    Mammalian P-glycoprotein is a highly conserved integral membrane protein functioning as an energy dependent plasma membrane efflux pump which decreases the concentration of certain lipophilic aromatic compounds entering the cell by diffusion. Studies indicate that P-glycoprotein is capable of increased expression in response to certain chemical stressors and has demonstrated the ability to transport xenobiotic contaminants. Expression of a xenobiotic transporter in teleost species could play a significant role in conferring resistance to fish populations exposed to xenobiotic stressors and may serve as a potential indicator of species at risk to environmental contaminants. Past studies demonstrated a strong correlation between corresponding mammalian and teleost tissues showing immunoreactivity to specific mammalian P-glycoprotein antibodies. In this study, comparisons of staining pattern, intensity, and tissue specificity between Lillie`s, Bouin`s and Dietrich`s fixed tissues was determined in the sheepshead minnow, Cyprinodon variegatus, using monoclonal antibodies (mAbs) C219, C494 and JSB-1. Immunoreactivity of the mAbs was found to be fixative-dependent and results are presented illustrating the differential staining patterns and tissue specificity observed for each tissue, fixative, and antibody combination. These data indicate tissue fixation has a significant impact on P-glycoprotein immunoreactivity in teleost tissues and must be considered in the comparison and interpretation of results.

  6. Quick radioimmunoassay for plasma immunoreactive gastrin. Application for localizing occult gastrinoma(s) during operation

    International Nuclear Information System (INIS)

    A rapid method for determining gastrin, quick gastrin, has been developed. Separation/washing procedure has been improved and can be completed within three minutes. It required only 48 minutes for the assay of 22 blood samples. Quick gastrin is a RIA that uses magnetic particles. On magnetic particles, a goat anti-rabbit IgG antibody is bound covalently. An anti-human gastrin rabbit antibody is bound to an anti-rabbit IgG antibody. Assay is started by adding the magnetic particles to a mixture of sample and 125I-gastrin. Following 30 minute incubation at 37degC, the particles are sedimented in a magnetic field and washed. The gastrin content of the sample is then quantitated by counting radioactivity of the particles. Incomplete equilibration of antigen-antibody reaction is corrected using standard solution prepared from charcoal treated plasma. The immunoreactive gastrin values by quick gastrin correlated well with those by a commercial assay kit (Gammadab RIA kit; y=1.01x+4.3, r=0.99). When compared to a reported conventional rapid assay, quick gastrin is easier and more accurate. Quick gastrin is sensitive enough to use for intra-operative determination of gastrin. We applied quick gastrin to the samples obtained from intra-operative secretin test in a gastrinoma patient. Twofold increase in gastrin after injection of secretin clearly indicated the existence of occult gastrinomas in her pancreas. When gastrin was assayed with the conventional rapid method, the increase in gastrin was less and did not reach the criteria for existence of gastrinoma. (author)

  7. Chronic 835-MHz radiofrequency exposure to mice hippocampus alters the distribution of calbindin and GFAP immunoreactivity. (United States)

    Maskey, Dhiraj; Pradhan, Jonu; Aryal, Bijay; Lee, Chang-Min; Choi, In-Young; Park, Ki-Sup; Kim, Seok Bae; Kim, Hyung Gun; Kim, Myeung Ju


    Exponential interindividual handling in wireless communication system has raised possible doubts in the biological aspects of radiofrequency (RF) exposure on human brain owing to its close proximity to the mobile phone. In the nervous system, calcium (Ca(2+)) plays a critical role in releasing neurotransmitters, generating action potential and membrane integrity. Alterations in intracellular Ca(2+) concentration trigger aberrant synaptic action or cause neuronal apoptosis, which may exert an influence on the cellular pathology for learning and memory in the hippocampus. Calcium binding proteins like calbindin D28-K (CB) is responsible for the maintaining and controlling Ca(2+) homeostasis. Therefore, in the present study, we investigated the effect of RF exposure on rat hippocampus at 835 MHz with low energy (specific absorption rate: SAR=1.6 W/kg) for 3 months by using both CB and glial fibrillary acidic protein (GFAP) specific antibodies by immunohistochemical method. Decrease in CB immunoreactivity (IR) was noted in exposed (E1.6) group with loss of interneurons and pyramidal cells in CA1 area and loss of granule cells. Also, an overall increase in GFAP IR was observed in the hippocampus of E1.6. By TUNEL assay, apoptotic cells were detected in the CA1, CA3 areas and dentate gyrus of hippocampus, which reflects that chronic RF exposure may affect the cell viability. In addition, the increase of GFAP IR due to RF exposure could be well suited with the feature of reactive astrocytosis, which is an abnormal increase in the number of astrocytes due to the loss of nearby neurons. Chronic RF exposure to the rat brain suggested that the decrease of CB IR accompanying apoptosis and increase of GFAP IR might be morphological parameters in the hippocampus damages. PMID:20546709

  8. Expression of dynamin immunoreactivity in experimental pancreatic tumors induced in rat by mancozeb-nitrosomethylurea. (United States)

    Valentich, M A; Cook, T; Urrutia, R


    Dynamins are GTPases which support receptor-mediated endocytosis and bind to several tyrosine kinase receptor-associated proteins known to mediate cell proliferation and differentiation. We have recently established that dynamin expression correlates with normal neuronal (Torre et al., J. Biol. Chem., 269 (1994) 32411-32417) and acinar pancreatic cell differentiation (Cook et al., Mol. Biol. Cell, 6 (1995) 405a). To begin to understand the role of dynamin in neoplastic pancreatic cell differentiation, we have followed the expression of this protein by immunohistochemistry during the development of pancreatic tumors in a mancozeb-nitrosomethylurea (NMU)-based carcinogenesis model recently developed in our laboratory (Monis and Valentich, Carcinogenesis, 14 (1993) 929-933). After a single intraperitoneal injection (50 mg/g body wt) of this carcinogen, rats fed with mancozeb develop pancreatic focal acinar hyperplasia (FACH), dysplastic foci (DYF) displaying acinar-like and ductular-like structures, and ductular-like carcinoma in situ (CIS). After histochemical staining using a monoclonal anti-dynamin antibody, high levels of this protein are consistently observed in well-differentiated acinar tumors (FACH). In contrast, dynamin immunoreactivity is almost undetectable in more advanced lesions showing a ductular-like phenotype (ductular-like DYF and CIS). This change in the expression pattern of dynamin during the progression of acinar into ductular-like DYF and CIS lesions correlates with recent findings from our laboratory showing a differential expression pattern for dynamin in pancreatic cells during embryonic development, with ductular-like precursor cells expressing low levels of this protein. Based upon these results, we conclude that more advanced ductular-like neoplastic cells induced by the carcinogen NMU in rat pancreas behave phenotypically like pancreatic precursor cells in their pattern of expression for dynamin. PMID:8603375

  9. Postnatal development of neuropeptide Y-like immunoreactivity in area 17 of normal and visually deprived rhesus monkeys. (United States)

    Tigges, M; Tigges, J; McDonald, J K; Slattery, M; Fernandes, A


    Immunocytochemical methods were used to examine neuropeptide Y (NPY) immunoreactive neurons and fibers in area 17 of rhesus monkeys during the first year of life. NPY-immunoreactive (+) neurons are nonpyramidal cells which are either multipolar, bipolar, or bitufted in shape. They occur most frequently in layer 6 and the subjacent white matter, are sparser in the supragranular layers, and absent from layer 4C. Labeled somata in the supragranular layers are smaller compared to those in layer 6 and the white matter. A typical axon originates from the NPY+ soma or from a primary dendrite and frequently is varicose. Distribution and morphologies of NPY+ neurons in area 17 of infants are similar to those of adult monkeys. Thus, it seems that NPY+ neurons in rhesus monkeys are mature from birth. NPY+ fibers occur in area 17 from birth; however, they differ in density and distribution from those of older infant and adult monkeys. At birth, a prominent fiber plexus is found in the deepest part of layer 1, and another in the white matter. Immunoreactive processes are sparse in the remaining cortical gray, except for some vertical fibers extending from pia to white matter. By 4 months of age, labeled fibers form a coarse network in layers 2, 3, 5, and 6. In addition, a distinct plexus extends through layers 4B, 4A, and the lowest aspect of layer 3. Also, a thin immunoreactive fiber band is found at the bottom of layer 4C. In the remainder of layer 4C, NPY+ fibers are scant. The supragranular layers also exhibit a unique immunoreactive "snarl" of fibers. Increases in density of NPY+ processes in the older infants are gradual so that between 7 and 13 months of age, NPY+ fibers appear to have achieved adultlike densities. These observations indicate that NPY+ fibers in area 17 of newborn rhesus monkeys undergo postnatal maturation which reaches a plateau around 4 months of age. After monocular visual deprivation from birth to 4 months of age, either by eyelid suture or by

  10. Changes in Brain Gonadotropin-Releasing Hormone- and Vasoactive Intestinal Polypeptide-like Immunoreactivity Accompanying Reestablishment of Photosensitivity in Male Dark-Eyed Juncos (Junco hyemalis)


    Deviche, Pierre; Saldanha, Colin J.; Silver, Rae


    In seasonally breeding, photoperiodic birds, the development of photorefractoriness is associated with decreased brain expression of gonadotropin-releasing hormone-like immunoreactivity (GnRH-li ir) and increased expression of vasoactive intestinal polypeptide-like immunoreactivity (VIP-li ir). Dissipation of photorefractoriness and reestablishment of photosensitivity are associated with increased GnRH-li ir brain production, but concurrent changes in VIP-li ir expression have not been invest...

  11. Cocaine induces striatal c-fos-immunoreactive proteins via dopaminergic D1 receptors.


    Young, S T; Porrino, L J; Iadarola, M. J.


    The protooncogene c-fos produces a phosphoprotein, Fos, which regulates gene transcription processes. In neuronal systems, Fos has been proposed to couple synaptic transmission to changes in gene expression by acting in the cell nucleus in concert with other proteins to form complexes in the promoter regions of target genes. We report here that the acute administration of a single dose of the indirect-acting dopaminergic agonist cocaine increases multiple Fos proteins in rat caudate nucleus. ...

  12. Calcium-binding Protein Calretinin Immunoreactivity in the Dog Superior Colliculus

    International Nuclear Information System (INIS)

    We studied calretinin-immunoreactive (IR) fibers and cells in the canine superior colliculus (SC) and studied the distribution and effect of enucleation on the distribution of this protein. Localization of calretinin was immunocytochemically observed. A dense plexus of anti-calretinin-IR fibers was found within the upper part of the superficial gray layer (SGL). Almost all of the labeled fibers were small in diameter with few varicosities. The intermediate and deep layers contained many calretinin-IR neurons. Labeled neurons within the intermediate gray layer (IGL) formed clusters in many sections. By contrast, labeled neurons in the deep gray layer (DGL) did not form clusters. Calretinin-IR neurons in the IGL and DGL varied in morphology and included round/oval, vertical fusiform, stellate, and horizontal neurons. Neurons with varicose dendrites were also labeled in the IGL. Most of the labeled neurons were small to medium in size. Monocular enucleation produced an almost complete reduction of calretinin-IR fibers in the SC contralateral to the enucleation. However, many calretinin-IR cells appeared in the contralateral superficial SC. Enucleation appeared to have no effect on the distribution of calretinin-IR neurons in the contralateral intermediate and deep layers of the SC. The calretinin-IR neurons in the superficial dog SC were heterogeneous small- to medium-sized neurons including round/oval, vertical fusiform, stellate, pyriform, and horizontal in shape. Two-color immunofluorescence revealed that no cells in the dog SC expressed both calretinin and GABA. Many horseradish peroxidase (HRP)-labeled retinal ganglion cells were seen after injections into the superficial layers. The vast majority of the double-labeled cells (HRP and calretinin) were small cells. The present results indicate that antibody to calretinin labels subpopulations of neurons in the dog SC, which do not express GABA. The results also suggest that the calretinin-IR afferents in the

  13. Celiac disease T cell epitopes from gamma-gliadins: immunoreactivity depends on the genome of origin, transcript frequency, and flanking protein variation

    NARCIS (Netherlands)

    Salentijn, E.M.J.; Mitea, D.C.; Goryunova, S.V.; Meer, van der I.M.; Padioleau, I.; Gilissen, L.J.W.J.; Koning, de F.; Smulders, M.J.M.


    Background - Celiac disease (CD) is caused by an uncontrolled immune response to gluten, a heterogeneous mixture of wheat storage proteins. The CD-toxicity of these proteins and their derived peptides is depending on the presence of specific T-cell epitopes (9-mer peptides; CD epitopes) that mediate

  14. Serologic immunoreactivity to Neospora caninum antigens in dogs determined by indirect immunofluorescence, western blotting and dot-ELISA. (United States)

    Pinheiro, A M; Costa, M F; Paule, B; Vale, V; Ribeiro, M; Nascimento, I; Schaer, R E; Almeida, M A O; Meyer, R; Freire, S M


    Neospora caninum, is a coccidian protozoan known as a major cause of bovine abortion and canine neuropathies. The aim of the present study was to develop a reliable and quick test to detect antibodies to N. caninum in dog sera. Sixty-five serum samples from dogs, including 35 positive and 30 negative for N. caninum antibodies were used for standardization of the test. In parallel, immunoreactivity of the sera to Toxoplasma gondii antigens was investigated using a passive agglutination test. A dot-ELISA test, using soluble extract of N. caninum tachyzoites on nitrocellulose ester membranes, was developed and standardized. SDS-PAGE and complementary analysis of reactivity by Western blotting were used for the characterization of the immunoreactive fractions of all tested sera. The sensitivity and specificity of the dot-ELISA were 94 and 73%, respectively, compared to IFAT at a cut-off of 1:50, and 87 and 100% compared to IFAT at a cut-off of 1:25. Among the sera that tested positively for both IFAT and dot-ELISA, only 8.6% were reactive to T. gondii. The most immunoreactive fractions in Western blots were the 14-, 33-, 42- and 55 kDa bands, with percentages of 42, 60, 42 and 37%, respectively. The 60 kDa band showed a non-specific reaction in 43% of neosporosis-negative animals by both dot-ELISA and IFAT. These results indicate that the dot-ELISA using N. caninum antigen present good sensitivity and specificity, and might be used as a screening test to detect antibodies to N. caninum in dogs. PMID:15893072

  15. Natural Immunoreactivity of Secretory IgA to Indigenous Strains of Streptococcus mutans From Chinese Spousal Pairs (United States)

    Nie, Min; Chen, Dong; Gao, Zhenyan; Wu, Xinyu; Li, Tong


    Background Dental caries is a well-known biofilm-mediated disease initiated by Streptococcus mutans, which should infect and colonize in a milieu perfused with components of the mucosal immune system. Little is known, however, regarding the relationship between the natural secretory IgA activity and S. mutans of a variety of diverse genotypes. Objectives The current study aimed to use spousal pairs to investigate the natural immunoreactivity of salivary secretory IgA to different genotype strains of S. mutans. Patients and Methods Indigenous strains were characterized from nine spousal pairs using polymerase reaction chain (PCR) and arbitrarily primed polymerase chain reaction (AP-PCR) by genotype monitoring. Unstimulated submandibular/sublingual secretions were collected and the concentrations of secretory IgA were determined by the enzyme-linked immunosorbent assay (ELISA). Each saliva sample was examined by Western blot to analyze the immunoreactivity of naturally occurring salivary secretory IgA antibodies for his/her own indigenous strain, spouse’s strain and reference strains including S. mutans GS-5 and Ingbritt (C). Results The results showed that naturally induced salivary IgA antibodies against S. mutans were present in all subjects. Almost all subjects had the similar individual immunoblotting profiles to different genotype strains. Conclusions The current study indicated that the immunoreactivity of secretory IgA might have no direct correlation with the colonization of indigenous flora and rejection of exogenous strains in adults. The relationship of microbes, host and dental caries should be in the light of coevolved microecosystem as a whole, but not caused by one factor alone. PMID:27303613

  16. Distribution and densitometry mapping of L1-CAM Immunoreactivity in the adult mouse brain – light microscopic observation

    Directory of Open Access Journals (Sweden)

    Yamasaki Hironobu


    Full Text Available Abstract Background The importance of L1 expression in the matured brain is suggested by physiological and behavioral studies showing that L1 is related to hippocampal plasticity and fear conditioning. The distribution of L1 in mouse brain might provide a basis for understanding its role in the brain. Results We examined the overall distribution of L1 in the adult mouse brain by immunohistochemistry using two polyclonal antibodies against different epitopes for L1. Immunoreactive L1 was widely but unevenly distributed from the olfactory bulb to the upper cervical cord. The accumulation of immunoreactive L1 was greatest in a non-neuronal element of the major fibre bundles, i.e. the lateral olfactory tract, olfactory and temporal limb of the anterior commissure, corpus callosum, stria terminalis, globus pallidus, fornix, mammillothalamic tract, solitary tract, and spinal tract of the trigeminal nerve. High to highest levels of non-neuronal and neuronal L1 were found in the grey matter; i.e. the piriform and entorhinal cortices, hypothalamus, reticular part of the substantia nigra, periaqueductal grey, trigeminal spinal nucleus etc. High to moderate density of neuronal L1 was found in the olfactory bulb, layer V of the cerebral cortex, amygdala, pontine grey, superior colliculi, cerebellar cortex, solitary tract nucleus etc. Only low to lowest levels of neuronal L1 were found in the hippocampus, grey matter in the caudate-putamen, thalamus, cerebellar nuclei etc. Conclusion L1 is widely and unevenly distributed in the matured mouse brain, where immunoreactivity was present not only in neuronal elements; axons, synapses and cell soma, but also in non-neuronal elements.

  17. Localisation of NG2 immunoreactive neuroglia cells in the rat locus coeruleus and their plasticity in response to stress

    Directory of Open Access Journals (Sweden)

    Mohsen eSeifi


    Full Text Available The locus coeruleus (LC nucleus modulates adaptive behavioural responses to stress and dysregulation of LC neuronal activity is implicated in stress-induced mental illnesses. The LC is composed primarily of noradrenergic neurons together with various glial populations. A neuroglia cell-type largely unexplored within the LC is the NG2 cell. NG2 cells serve primarily as oligodendrocyte precursor cells throughout the brain. However, some NG2 cells are in synaptic contact with neurons suggesting a role in information processing. The aim of this study was to neurochemically and anatomically characterise NG2 cells within the rat LC. Furthermore, since NG2 cells have been shown to proliferate in response to traumatic brain injury, we investigated whether such NG2 cells plasticity also occurs in response to emotive insults such as stress. Immunohistochemistry and confocal microscopy revealed that NG2 cells were enriched within the pontine region occupied by the LC. Close inspection revealed that a sub-population of NG2 cells were located within unique indentations of LC noradrenergic somata and were immunoreactive for the neuronal marker NeuN whilst NG2 cell processes formed close appositions with clusters immunoreactive for the inhibitory synaptic marker proteins gephyrin and the GABA-A receptor alpha3-subunit, on noradrenergic dendrites. In addition, LC NG2 cell processes were decorated with vesicular glutamate transporter 2 immunoreactive puncta. Finally, ten days of repeated restraint stress significantly increased the density of NG2 cells within the LC. The study demonstrates that NG2 IR cells are integral components of the LC cellular network and they exhibit plasticity as a result of emotive challenges.

  18. Changes in small intestinal chromogranin A-immunoreactive cell densities in patients with irritable bowel syndrome after receiving dietary guidance. (United States)

    Mazzawi, Tarek; El-Salhy, Magdy


    Chromogranin A (CgA) is a common marker for enteroendocrine cells in the gut, and CgA-immunoreactive cell densities are abnormal in patients with irritable bowel syndrome (IBS). The majority of patients with IBS report that their symptoms develop after consuming certain foodstuffs. In the present study, we investigated the effects of dietary guidance on the total enteroendocrine cell densities in the small intestine, as detected by CgA. A total of 14 patients with IBS underwent a gastroscopy with duodenal biopsies and 11 of them also underwent a colonoscopy, with biopsy samples obtained from the ileum. Fourteen control subjects were also included. Each patient received 3 sessions of dietary guidance. Gastroscopies and colonoscopies were performed on both the controls and patients with IBS (at baseline and at 3-9 months after receiving guidance). Biopsy samples obtained from the duodenum and ileum were immunostained for CgA using the avidin-biotin complex (ABC) method and were quantified using computerized image analysis. The density of CgA-immunoreactive cells in the duodenum (mean ± SEM values) in the control subjects was 235.9 ± 31.9 cells/mm2; in the patients with IBS, the density was 36.9 ± 9.8 and 103.7 ± 16.9 cells/mm2 before and after they received dietary guidance, respectively (P=0.007). The density of CgA-immunoreactive cells in the ileum in the control subjects was 47.4 ± 8.3 cells/mm2; in the patients with IBS, the density was 48.4 ± 8.1 and 17.9 ± 4.4 cells/mm2, before and after they received dietary guidance, respectively (P=0.0006). These data indicate that changes in CgA-immunoreactive cell densities in patients with IBS after receiving dietary guidance may reflect a change in the densities of the small intestinal enteroendocrine cells, which may contribute to an improvement in the IBS symptoms. PMID:26987104

  19. Loss of parvalbumin-immunoreactivity in mouse brain regions after repeated intermittent administration of esketamine, but not R-ketamine. (United States)

    Yang, Chun; Han, Mei; Zhang, Ji-Chun; Ren, Qian; Hashimoto, Kenji


    Clinical use of the rapid antidepressant drug ketamine is limited, due to psychotomimetic side effects. R-ketamine appears to be a potent, long-lasting and safer antidepressant, relative to S-ketamine (esketamine), since it is free of psychotomimetic side effects. Repeated, intermittent administration of esketamine (10mg/kg, once per week for 8-weeks), but not R-ketamine, caused loss of parvalbumin (PV)-immunoreactivity in the medial prefrontal cortex and hippocampus of mouse brains, regions associated with psychosis. This study suggests that repeated intermittent use of R-ketamine is safer than esketamine in the treatment of depression. PMID:27043274

  20. Expression of Helicobacter pylori hspA Gene in Lactococcus lactis NICE System and Experimental Study on Its Immunoreactivity

    Directory of Open Access Journals (Sweden)

    Xiao-Juan Zhang


    Full Text Available Aim. The aim of this study was to develop an oral Lactococcus lactis (L. lactis vaccine against Helicobacter pylori (H. pylori. Methods. After L. lactis NZ3900/pNZ8110-hspA was constructed, growth curves were plotted to study whether the growth of recombinant L. lactis was affected after hspA was cloned into L. lactis and whether the growth of empty bacteria, empty plasmid bacteria, and recombinant L. lactis was affected by different concentrations of Nisin; SDS-PAGE and Western blot were adopted, respectively, to detect the HspA expressed by recombinant L. lactis and its immunoreactivity. Results. There was no effect observed from the growth curve after exogenous gene hspA was cloned into L. lactis NZ3900; different concentrations of Nisin did not affect the growth of NZ3900 and NZ3900/pNZ8110, while different concentrations of Nisin inhibited the growth of NZ3900/pNZ8110-hspA except 10 ng/mL Nisin. No HspA strip was observed from SDS-PAGE. Western blot analysis showed that HspA expressed by recombinant bacteria had favorable immunoreactivity. Conclusion. The growth of recombinant L. lactis was suppressed even though a small amount of HspA had been induced to express. Therefore recombinant L. lactis only express HspA which was not suitable to be oral vaccine against Helicobacter pylori.

  1. Absent and abundant MET immunoreactivity is associated with poor prognosis of patients with oral and oropharyngeal squamous cell carcinoma. (United States)

    De Herdt, Maria J; Willems, Stefan M; van der Steen, Berdine; Noorlag, Rob; Verhoef, Esther I; van Leenders, Geert J L H; van Es, Robert J J; KoljenoviÄ, Senada; Baatenburg de Jong, Robert J; Looijenga, Leendert H J


    Although the receptor tyrosine kinase (RTK) MET is widely expressed in head and neck squamous cell carcinoma (HNSCC), its prognostic value remains unclear. This might be due to the use of a variety of antibodies and scoring systems. Here, the reliability of five commercial C-terminal MET antibodies (D1C2, CVD13, SP44, C-12 and C-28) was evaluated before examining the prognostic value of MET immunoreactivity in HNSCC. Using cancer cell lines, it was shown that D1C2 and CVD13 specifically detect MET under reducing, native and formalin-fixed paraffin-embedded (FFPE) conditions. Immunohistochemical staining of routinely FFPE oral SCC with D1C2 and CVD13 demonstrated that D1C2 is most sensitive in the detection of membranous MET. Examination of membranous D1C2 immunoreactivity with 179 FFPE oral and oropharyngeal SCC - represented in a tissue microarray - illustrated that staining is either uniform (negative or positive) across tumors or differs between a tumor's center and periphery. Ultimately, statistical analysis revealed that D1C2 uniform staining is significantly associated with poor 5-year overall and disease free survival of patients lacking vasoinvasive growth (HR = 3.019, p < 0.001; HR = 2.559, p < 0.001). These findings might contribute to reliable stratification of patients eligible for treatment with biologicals directed against MET. PMID:26909606

  2. Immunoreactivity of glucose transporter 8 is localized in the epithelial cells of the choroid plexus and in ependymal cells. (United States)

    Murakami, Ryuta; Chiba, Yoichi; Tsuboi, Kazuhito; Matsumoto, Koichi; Kawauchi, Machi; Fujihara, Ryuji; Mashima, Masato; Kanenishi, Kenji; Yamamoto, Tetsuji; Ueno, Masaki


    High fructose intake is known to be associated with increased plasma triglyceride concentration, impaired glucose tolerance, insulin resistance, and high blood pressure. In addition, excess fructose intake is also thought to be a risk factor for dementia. Previous immunohistochemical studies have shown the presence of glucose transporter 5 (GLUT5), a major transporter of fructose, in the epithelial cells of the choroid plexus and ependymal cells in the brains of humans, rats, and mice, while GLUT2, a minor transporter of fructose, was localized in the ependymal cells of rat brain. In this study, immunoreactivity for the fructose transporter GLUT8 was observed in the cytoplasm of the epithelial cells in the choroid plexus and in the ependymal cells of the brains of humans and mice. These structures were not immunoreactive for GLUT7, GLUT11, and GLUT12. Our findings support the hypothesis of the transport of intravascular fructose through the epithelial cells of the choroid plexus and the ependymal cells. PMID:27160096

  3. Noradrenaline as a putative neurotransmitter mediating hypotension—induced FOs—like immunoreactivity in the supraoptic nucleus of the rat

    Institute of Scientific and Technical Information of China (English)



    Hemorrhage or hypotension induces extensive Fos-like immunoreactivity in the magnocellular neurosecretory cells in the supraoptic nucleus of the hypothalamus in rat,especially in the vasopressin neurons.The present study was to explore the neurotransmitter mediating this effect,Microinfusion of the alpha-adrenergic blocker into the supraoptic nucleus reduced the hypotension-induced FOs.whereas beta-antagonist did not affect it significantly.Alaha1-and alpha2-antagonist,prazosin and yohimbine,both reduced the Fos-Positive cell counts.However,the effective dosage of yohimbine was much larger,Alpha1-agonist,methoxamine,induced abundant Fos-like immunoreactivity in the vasopressin cells in this nucleus,while beta-and alpha2-agonist did not elicit such effect.Administration of the noradrenergic re-uptake inhibitor desipramine,to this nucleus to locally accumulate the spontaneously released noradrenaline from the nerve terminals also induced Fos expression,mostly in the vasopressin cells.

  4. Serotonin-like immunoreactivity in the central and peripheral nervous systems of the interstitial acochlidean Asperspina sp. (Opisthobranchia). (United States)

    Hochberg, Rick


    Species of Acochlidea are common members of the marine interstitial environment and defined in part by their minuscule size and highly divergent morphology relative to other benthic opisthobranchs. Despite these differences, acochlideans such as species of Asperspina display many plesiomorphic characteristics, including an unfused condition of their neural ganglia. To gain insight into the distribution of specific neural subsets within acochlidean ganglia, a species of Asperspina was studied by using anti-serotonin immunohistochemistry and epifluorescence and confocal laser scanning microscopy. Results reveal similarities between Asperspina and larger opisthobranchs in the general distribution of serotonergic perikarya in the central nervous system. Specifically, the arrangement of perikarya into regional clusters within the cerebral and pedal ganglia and the absence of immunoreactive perikarya in the pleural ganglia are similar to the model species of Aplysia californica, Pleurobranchaea californica, and Tritonia diomedea. Moreover, serotonergic innervation of the rhinophores in all opisthobranchs, including Asperspina sp., originates from the cerebral ganglion instead of directly from the rhinophoral ganglion. Serotonergic innervation of the body wall, including the epithelium, muscles, and pedal sole, appears to arise exclusively from pedal and accessory ganglia. These observations indicate a general conservation of serotonin-like immunoreactivity in the central and peripheral nervous systems of acochlidean and other benthic opisthobranchs. PMID:17679719

  5. Bayesian Music Transcription

    NARCIS (Netherlands)

    Cemgil, A.T.


    Music transcription refers to extraction of a human readable and interpretable description from a recording of a music performance. The final goal is to implement a program that can automatically infer a musical notation that lists the pitch levels of notes and corresponding score positions in any a

  6. Stereologic estimates of total spinophilin-immunoreactive spine number in area 9 and the CA1 field: relationship with the progression of Alzheimer’s disease (United States)

    Akram, Afia; Christoffel, Daniel; Rocher, Anne B.; Bouras, Constantin; Kövari, Enikö; Perl, Daniel P.; Morrison, John H.; Herrmann, François R.; Haroutunian, Vahram; Giannakopoulos, Panteleimon; Hof, Patrick R.


    The loss of presynaptic markers is thought to represent a strong pathologic correlate of cognitive decline in Alzheimer’s disease (AD). Spinophilin is a postsynaptic marker mainly located to the heads of dendritic spines. We assessed total numbers of spinophilin-immunoreactive puncta in the CA1 and CA3 fields of hippocampus and area 9 in 18 elderly individuals with various degrees of cognitive decline. The decrease in spinophilin-immunoreactivity was significantly related to both Braak neurofibrillary tangle (NFT) staging and clinical severity but not Aβ deposition staging. The total number of spinophilin-immunoreactive puncta in CA1 field and area 9 were significantly related to MMSE scores and predicted 23.5% and 61.9% of its variability. The relationship between total number of spinophilin-immunoreactive puncta in CA1 field and MMSE scores did not persist when adjusting for Braak NFT staging. In contrast, the total number of spinophilin-immunoreactive puncta in area 9 was still significantly related to the cognitive outcome explaining an extra 9.6% of MMSE and 25.6% of the Clinical Dementia Rating scores variability. Our data suggest that neocortical dendritic spine loss is an independent parameter to consider in AD clinicopathologic correlations. PMID:17420070

  7. Changes in oxytocin immunoreactivity and mRNA expression in the sheep brain during pregnancy, parturition and lactation and in response to oestrogen and progesterone. (United States)

    Broad, K D; Kendrick, K M; Sirinathsinghji, D J; Keverne, E B


    The effects of pregnancy, parturition and lactation and exogenous treatments with oestradiol and progesterone on oxytocin (OXY) immunoreactivity and gene expression in the sheep brain were investigated. Immunocytochemistry was used to demonstrate that increased OXY-immunoreactivity occurred in cells of the paraventricular (PVN) and supraoptic nuclei (SON), the bed nucleus of the stria terminalis (BNST), the anterior commissural nuclei (ACN) and the periventricular part of the medial preoptic area (PvMP). Oxytocin immunoreactive terminals were also seen in the accessory olfactory nucleus, the glomerular and peri-glomerular layers of the olfactory bulb, the lateral septum, the zona incerta and the pars compacta of the substantia nigra. Compared to ovariectomized and late pregnant animals, the intensity of immunoreactivity was increased in all of these oxytocinergic elements at parturition, during lactation and following exogenous treatment with oestradiol. The OXY-immunoreactivity was also more intense in late pregnant animals compared to ovariectomized ones. Quantitative in situ hybridization histochemistry showed that cells in the PVN, SON, BNST and PvMP all showed significantly increased expression of OXY mRNA in animals at parturition and during lactation compared to late pregnant or ovariectomized animals. Expression levels in late pregnant animals were also significantly higher than in ovariectomized ones. Progesterone treatment significantly increased OXY mRNA in the PVN, SON, BNST and PvMP whereas oestradiol treatment was only effective in the PVN, BNST and PvMP. Combined treatment with these steroids did not significantly increase OXY mRNA levels in comparison with their administration alone. These results show that OXY-immunoreactivity and mRNA expression are at their highest in the sheep brain when maternal behaviour is induced. The increased synthesis/storage of the peptide at parturition may be due to changes in circulating concentrations of both

  8. DNA Topoisomerases in Transcription

    DEFF Research Database (Denmark)

    Rødgaard, Morten Terpager


    This Ph.D. thesis summarizes the main results of my studies on the interplay between DNA topoisomerases and transcription. The work was performed from 2011 to 2015 at Aarhus University in the Laboratory of Genome Research, and was supervised by associate professor Anni H. Andersen. Most of the ex......This Ph.D. thesis summarizes the main results of my studies on the interplay between DNA topoisomerases and transcription. The work was performed from 2011 to 2015 at Aarhus University in the Laboratory of Genome Research, and was supervised by associate professor Anni H. Andersen. Most...... topoisomerase-DNA cleavage complex. The second study is an investigation of how topoisomerases influence gene regulation by keeping the genome in an optimal topological state....

  9. Dopamine D1 and D2 receptor immunoreactivities in the arcuate-median eminence complex and their link to the tubero-infundibular dopamine neurons

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    W. Romero-Fernandez


    Full Text Available Dopamine D1 and D2 receptor immunohistochemistry and Golgi techniques were used to study the structure of the adult rat arcuate-median eminence complex, and determine the distribution of the dopamine D1 and D2 receptor immunoreactivities therein, particularly in relation to the tubero-infundibular dopamine neurons. Punctate dopamine D1 and D2 receptor immunoreactivities, likely located on nerve terminals, were enriched in the lateral palisade zone built up of nerve terminals, while the densities were low to modest in the medial palisade zone. A codistribution of dopamine D1 receptor or dopamine D2 receptor immunoreactive puncta with tyrosine hydroxylase immunoreactive nerve terminals was demonstrated in the external layer. Dopamine D1 receptor but not dopamine D2 receptor immnunoreactivites nerve cell bodies were found in the ventromedial part of the arcuate nucleus and in the lateral part of the internal layer of the median eminence forming a continuous cell mass presumably representing neuropeptide Y immunoreactive nerve cell bodies. The major arcuate dopamine/ tyrosine hydroxylase nerve cell group was found in the dorsomedial part. A large number of tyrosine hydroxylase immunoreactive nerve cell bodies in this region demonstrated punctate dopamine D1 receptor immunoreactivity but only a few presented dopamine D2 receptor immunoreactivity which were mainly found in a substantial number of tyrosine hydroxylase cell bodies of the ventral periventricular hypothalamic nucleus, also belonging to the tubero-infundibular dopamine neurons. Structural evidence for projections of the arcuate nerve cells into the median eminence was also obtained. Distal axons formed horizontal axons in the internal layer issuing a variable number of collaterals classified into single or multiple strands located in the external layer increasing our understanding of the dopamine nerve terminal networks in this region.  Dopamine D1 and D2 receptors may therefore directly

  10. Radioimmunoassay of digoxin in serum using monoclonal antibodies and assessment of interference by digoxin-like immunoreactive substances

    International Nuclear Information System (INIS)

    We used 7 monoclonal antibodies (MoAbs) and one polyclonal antibody to develop radioimmunoassays (RIAs) for digoxin in serum or plasma. These RIAs were tested for measuring apparent digoxin concentrations in serum from patients receiving the drug, from normal individuals, and in cord blood plasma. We found that two MoAbs cross-reacted significantly with substances in cord blood. The magnitude of cross-reactivity was dependent on the incubation time and temperature. Under equilibrium conditions, one antibody gave apparent digoxin values in cord blood plasma averaging 2.15 ng/ml. We suggest that this cross-reactivity is partially due to progesterone and 17-hydroxyprogesterone in cord blood plasma. The antibody that shows high cross-reactivity with digoxin-like immunoreactive substances may prove a useful tool for studies dealing with characterization of the cross-reacting compounds

  11. The effect of repeated stress on KCC2 and NKCC1 immunoreactivity in the hippocampus of female mice

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    Takao Tsukahara


    The data were obtained from the hippocampus of female mice using single-plane confocal microscopy images. The mean fluorescence intensity of the perisomatic area of neurons, defined as raw fluorescence intensity (RFI was calculated. Repeated stress (RS resulted in a decrease in perisomatic area of immunoreactive (IR-KCC2 and an increase of the IR-NKCC1. In addition, RS decreased perisomatic IR-pKCC2ser940, corresponding to that of KCC2. The data in this article support the results of a previous study [1] and provide the details of immunohistological methods. Interpretation of the data in this article can be found in “Repeated stress-induced expression pattern alterations of the hippocampal chloride transporters KCC2 and NKCC1 associated with behavioral abnormalities in female mice” by Tsukahara et al. [1].

  12. Lysine-directed conjugation of ethidium homodimer to B72.3 antibody: retention of immunoreactivity but altered tumor targeting

    International Nuclear Information System (INIS)

    Ethidium homodimer (EHD) was conjugated to B72.3 monoclonal antibody using a method whereby 85-90% of the conjugated EHD remains available for DNA intercalation. Antibody was thiopropionylated by reaction with N-succinimidyl 3-(2-pyridyldithio)propionate and reduction of pyridyldithio groups with dithiothreitol. EHD was maleimido-functionalized with succinimidyl-4-(N-maleimidoethyl)cyclohexane-1-carboxylate and treated with thiopropionylated antibody to obtain a conjugate containing ∼3.4 EHD per antibody molecule. For biologic studies, 14C-labeled EHD was synthesized by reductive amination and conjugated as above. In vitro the conjugate maintained chemical integrity and immunoreactivity, while in vivo its targeting of LS174T tumors was reduced compared with that of iodinated antibody. A decrease in isoelectric point of the immunoconjugate was also observed

  13. The rate of rise of corticotrophin releasing factor and endogenous digoxin-like immunoreactivity in normal and abnormal pregnancy. (United States)

    Wolfe, C D; Petruckevitch, A; Quartero, R; Carabelli, P; Poston, L; Kerkez, S; Campbell, E; Lowry, P J; Linton, E A


    Maternal plasma concentrations of corticotrophin releasing factor (CRF) and endogenous digoxin-like immunoreactivity (EDLI) were estimated in 80 normal and 88 abnormal pregnancies which were sampled sequentially from 24 weeks gestation to delivery. A slope was fitted for each woman's antenatal EDLI and CRF values, both of which rose significantly during gestation, and the mean of the slopes for the normal and abnormal groups for each value compared. There was no evidence of significant mean differences between groups for EDLI but there was evidence of a significant mean difference for CRF (P less than 0.05). After adjustment for other variables which may affect pregnancy outcome, the slopes for CRF were found not to be significantly related to outcome. PMID:2242370

  14. Transamidation of gluten proteins during the bread-making process of wheat flour to produce breads with less immunoreactive gluten. (United States)

    Heredia-Sandoval, Nina Gisella; Islas-Rubio, Alma Rosa; Cabrera-Chávez, Francisco; Calderón de la Barca, Ana María


    Due to an increasing incidence of celiac disease (CD) and other gluten-related disorders, different gluten-free breads have been developed using starches and additives as a substitute for gluten. Thus, patients miss not only the taste and aroma of wheat bread but also risk their sensitive intestines. Therefore, modifying gluten to avoid an immune response in CD and its application to baking is in progress. The aim of the study was to enzymatically modify gluten on wheat flour, during bread-making avoiding the use of additives, to reduce immunoreactivity, preserving its properties. Microbial transglutaminase (mTG) or chymotrypsin (ChT) was used to bind lysine or valine to gluten proteins in a model system. The best conditions were directly applied to wheat flour for bread-making with and without punching at 45 min. Subsequently, the rheological properties of the doughs, specific volume of the loaves, immunoreactive gluten content and modification of the extracted proteins were evaluated. ChT-treated breads presented a better appearance with a more homogeneous crumb, higher specific volume values (3.34-4.25 cm(3) g(-1)) and higher reactive gluten reduction (up to 71%) than the mTG-treated ones (1.23-2.66 cm(3) g(-1)) with only a 42% reactive gluten reduction. Thus, transpeptidation during bread-making is a promising technology, although it is necessary to improve the modification process to obtain the reactive gluten reduction required in breads for the treatment of CD patients and other gluten-related disorders. PMID:24917417

  15. Reduced density of glutamine synthetase immunoreactive astrocytes in different cortical areas in major depression but not in bipolar I disorder

    Directory of Open Access Journals (Sweden)

    Hans-Gert eBernstein


    Full Text Available There is increasing evidence for disturbances within the glutamate system in patients with affective disorders, which involve disruptions of the glutamate-glutamine- cycle. The mainly astroglia-located enzyme glutamine synthetase catalyzes the ATP-dependent condensation of ammonia and glutamate to form glutamine, thus playing a central role in glutamate and glutamine homoeostasis. However, glutamine synthetase is also expressed in numerous oligodendrocytes, another class of glial cells implicated in mood disorder pathology. To learn more about the role of glia-associated glutamine synthetase in mental illnesses, we decided to find out if numerical densities of glial cells immunostained for the enzyme protein differ between subjects with major depressive disorder, bipolar disorder and psychically healthy control cases. Counting of glutamine synthetase expressing astrocytes and oligodendrocytes in eight cortical and two subcortical brain regions of subjects with mood disorder (N=14, bipolar disorder (N=15 and controls (N=16 revealed that in major depression the densities of astrocytes were significantly reduced in some cortical but not subcortical gray matter areas, whereas no changes were found for oligodendrocytes. In bipolar disorder no alterations of glutamine synthetase-immunoreactive glia were found. From our findings we conclude that (1 glutamine synthetase expressing astrocytes are prominently involved in glutamate-related disturbances in major depression, but not in bipolar disorder and (2 glutamine synthetase expressing oligodendrocytes, though being present in significant numbers in prefrontal cortical areas, play a minor (if any role in mood disorder pathology. The latter assumption is supported by findings of others showing that - at least in the mouse brain cortex - glutamine synthetase immunoreactive oligodendroglial cells are unable to contribute to the glutamate-glutamine cycle due to the complete lack of amino acid transporters

  16. Effects of food deprivation on goal-directed behavior, spontaneous locomotion, and c-Fos immunoreactivity in the amygdala. (United States)

    Moscarello, J M; Ben-Shahar, O; Ettenberg, A


    Previous work in our laboratory has shown that food deprivation and food presentation produce different patterns of neuronal activity (as measured by c-Fos immunoreactivity) in the medial prefrontal cortex and nucleus accumbens of rats. Since the amygdala has been implicated in both motivational and reinforcement processes and has neuronal connections to both the prefrontal cortex and nucleus accumbens, it was of interest to assess amygdaloid c-Fos immunoreactivity during similar manipulations of food deprivation and presentation. In the current study, c-Fos counts in both basolateral and central amygdalar nuclei were observed to increase in rats 12- and 36-h food deprived (relative to 0-h controls)-an effect reversed by the presentation of either a small or large meal (2.5 or 20g of food). In another experiment, rats working on a progressive ratio schedule of reinforcement exhibited elevated break-points as a function of food deprivation, a result consistent with the view that the feeding manipulations increased the subjects' level of motivation. In contrast, food deprivation reduced the spontaneous locomotor activity of rats, presumably as a result of an inherent energy-conservation strategy when no food is readily available. These data suggest that the state of food deprivation is associated with: (a) enhanced behavioral output only when food is attainable (increased goal-directed behavior, but decreased spontaneous activity), and (b) increased synaptic engagement in neuronal circuits involved in affective valuation and related decision-making (increased c-Fos counts in the amygdala). PMID:18706934

  17. Androgen inhibits the increases in hypothalamic corticotropin-releasing hormone (CRH) and CRH-immunoreactivity following gonadectomy. (United States)

    Bingaman, E W; Magnuson, D J; Gray, T S; Handa, R J


    To characterize the effect of androgens on the hypothalamo-pituitary-adrenal (HPA) axis we examined the regulation of corticotropin-releasing hormone (CRH) following gonadectomy and hormone replacement. Three-month-old male Fischer 344 (F344) rats were gonadectomized (GDX) or sham GDX. Control animals remained intact. Animals were sacrificed 1, 4, 7, 10, or 21 days following surgery. GDX rats had significantly elevated (p < 0.05) levels of hypothalamic CRH 21 days after surgery compared to intact and sham-operated rats. In a second study, 3-month-old male F344 rats were GDX and treated with the non-aromatizable androgen, dihydrotestosterone (DHT), using a Silastic capsule containing crystalline DHT propionate subcutaneously implanted in each animal's back. Control animals were GDX and sham-treated or left intact (INT). Three weeks following gonadectomy, CRH levels in the hypothalamus of GDX rats showed a significant increase (p < 0.05) compared to intact animals. DHT treatment, beginning at the time of gonadectomy prevented this increase. CRH or arginine vasopressin (AVP) immunoreactivity was examined using immunocytochemistry. The number of CRH-immunoreactive (IR) cells in the paraventricular nucleus (PVN) of GDX, DHT-treated animals was significantly decreased (p < 0.05) compared to GDX rats. No differences were seen between treatment groups in CRH-IR cell numbers in the bed nucleus of the stria terminalis or the central amygdaloid nucleus or in AVP-IR cell numbers in the PVN. These data demonstrate that long-term castration increases hypothalamic CRH content and CRH-IR cell numbers in the PVN by removal of an androgen-dependent repression. PMID:8159272

  18. Development and distribution of PAG-immunoreactive neurons in the central pathway of trigeminal proprioception of the rat brainstem

    Institute of Scientific and Technical Information of China (English)

    PANG You-wang; LI Jin-lian


    Objective:To investigate the development and distribution of phosphate-activated glutaminase like immunoreactive (PAG-LI) neurons in the central pathway of trigeminal proprioception of the rat brainstem.Methods: The immunohistochemitry techniques were used. Results: (1) At embryonic day 17 (E17), PAGLI neurons were initially observed in the mesencephalic trigeminal nucleus (Vme). All PAG-LI neurons were large round neurons with moderate immunostaining. The immunoreactivity grew intense and attained adultlike pattern at P10. (2) Not until postnatal day 10 (P10) did a few PAG-LI neurons appear in the area ventral to the motor trigeminal nucleus (AVM) and area dorsal to the superior olivery nucleus (ADO), and not until P12 in the dorsomedial part of the subnucleus oralis of the spinal trigeminal nucleus (Vodm) and dorsomedial part of the principal sensory trigeminal nucleus (Vpdm). As development proceeded, more and more neurons in them were immunostained, and some PAG-LI neurons were detected in the lateral reticular formation adjacent to the Vodm(LRF)and the caudolateral part of the supratrigeminal nucleus (Vsup-CL) at P21.Conclusion: In the central pathway of trigeminal proprioception of the rat brainstem, PAG-LI neurons appeared during two stages: The first stage from E17 to P10, PAG-LI neurons appeared in the Vme and reached adult-like pattern; the second stage from P10 to P21, PAG-LI neurons appeared in the Vodm, LRF,Vpdm, Vsup-CL, ADO, AVM and gradually reached adult-like pattern. This might be relative to the establishment of jaw movement patterns.

  19. Effects of sex and reproductive experience on the number of orexin A-immunoreactive cells in the prairie vole brain. (United States)

    Donlin, Michael; Cavanaugh, Breyanna L; Spagnuolo, Olivia S; Yan, Lily; Lonstein, Joseph S


    Large populations of cells synthesizing the neuropeptide orexin (OX) exist in the caudal hypothalamus of all species examined and are implicated in physiological and behavioral processes including arousal, stress, anxiety and depression, reproduction, and goal-directed behaviors. Hypothalamic OX expression is sexually dimorphic in different directions in laboratory rats (F>M) and mice (M>F), suggesting different roles in male and female physiology and behavior that are species-specific. We here examined if the number of hypothalamic cells immunoreactive for orexin A (OXA) differs between male and female prairie voles (Microtus ochrogaster), a socially monogamous species that pairbonds after mating and in which both sexes care for offspring, and if reproductive experience influences their number of OXA-immunoreactive (OXA-ir) cells. It was found that the total number of OXA-ir cells did not differ between the sexes, but females had more OXA-ir cells than males in anterior levels of the caudal hypothalamus, while males had more OXA-ir cells posteriorly. Sexually experienced females sacrificed 12 days after the birth of their first litter, or one day after birth of a second litter, had more OXA-ir cells in anterior levels but not posterior levels of the caudal hypothalamus compared to females housed with a brother (incest avoidance prevents sibling mating). Male prairie voles showed no effect of reproductive experience but showed an unexpected effect of cohabitation duration regardless of mating. The sex difference in the distribution of OXA-ir cells, and their increased number in anterior levels of the caudal hypothalamus of reproductively experienced female prairie voles, may reflect a sex-specific mechanism involved in pairbonding, parenting, or lactation in this species. PMID:24874707

  20. Significant difference in p53 and p21 protein immunoreactivity in HPV 16 positive and HPV negative breast carcinomas

    International Nuclear Information System (INIS)

    Human papillomavirus (HPV) 16 has previously been found in 19/41 breast carcinomas (46%) in women with a history of HPV 16 positive CIN III lesions. There was no significant difference in distribution of histological subtypes, mean or median tumour diameter or number of regional lymph node metastases in the HPV positive and HPV negative breast carcinoma groups. P53, p21 and c-erbB-2 proteins were analyzed by immunohistochemistry in the HPV 16 positive and HPV negative breast carcinomas. There was a significant difference in p53 and p21 protein immunoreactivity between HPV 16 positive and HPV negative breast carcinomas (p=0.0091 and p=0.0040), with a significant less detectable p53 and p21 protein immunoreactivity in the HPV 16 positive cases. There was also a significant difference in the coexpression of p53/p21 between the HPV 16 positive and HPV 16 negative breast carcinomas (p=0.002). No significant difference in immunostaining for c-erbB-2 protein in the two groups was found (p=0.15), or for the coexpression of p53/c-erbB-2 (p=0.19). The significantly lower expression of p53 and p21 proteins in HPV 16 positive than in HPV 16 negative breast carcinomas supports the hypothesis of inactivation and degradation of wild-type p53 proteins by HPV 16 E6 and that p53 mutation is not necessary for transformation in the HPV 16 positive cases. (orig.)

  1. VEGF, VEGFR-1 and VEGFR-2 immunoreactivity in the porcine arteries of vascular subovarian plexus (VSP during the estrous cycle.

    Directory of Open Access Journals (Sweden)

    A Andronowska


    Full Text Available Abstract: Vascular endothelial growth factor (VEGF is an important angiogenic factor in the female reproductive tract. It binds to cell surface through ligand-stimulatable tyrosine kinase receptors, the most important being VEGFR-1 (flt-1 and VEGFR-2 (flk-1. The broad ligament of the uterus is a dynamic organ consisting of specialized complexes of blood vessels connected functionally to the uterus, oviduct and ovary. Endothelial cells form an inner coating of the vessel walls and thus they stay under the influence of various modulators circulating in blood including ovarian steriods involved in developmental changes in the female reproductive system. The aim of the present study was to immunolocalize VEGF and its two receptors: VEGFR-1 and VEGFR-2 in the broad ligament of the uterus in the area of vascular subovarian plexus during different phases of the estrous cycle in pig and to determine the correlation between immunoreactivity of the investigated factors and phases of the estrous cycle. The study was performed on cryostat sections of vascular subovarian plexus stained immunohistochemically by ABC method. Specific polyclonal antibodies: anti-VEGF, anti-VEGFR-1 and anti-VEGFR-2 were used. Data were subjected to one-way analysis of variance. Our study revealed the presence of VEGF and its receptors in endothelial and smooth muscle cells of VSP arteries. All agents displayed phase-related differences in immunoreactivity suggesting the modulatory effect of VEGF, VEGFR-1 and VEGFR-2 on the arteries of the VSP in the porcine broad ligament of the uterus.

  2. Ubiquitin and proteasomes in transcription. (United States)

    Geng, Fuqiang; Wenzel, Sabine; Tansey, William P


    Regulation of gene transcription is vitally important for the maintenance of normal cellular homeostasis. Failure to correctly regulate gene expression, or to deal with problems that arise during the transcription process, can lead to cellular catastrophe and disease. One of the ways cells cope with the challenges of transcription is by making extensive use of the proteolytic and nonproteolytic activities of the ubiquitin-proteasome system (UPS). Here, we review recent evidence showing deep mechanistic connections between the transcription and ubiquitin-proteasome systems. Our goal is to leave the reader with a sense that just about every step in transcription-from transcription initiation through to export of mRNA from the nucleus-is influenced by the UPS and that all major arms of the system--from the first step in ubiquitin (Ub) conjugation through to the proteasome-are recruited into transcriptional processes to provide regulation, directionality, and deconstructive power. PMID:22404630

  3. Conserved hemopoietic transcription factor Cg-SCL delineates hematopoiesis of Pacific oyster Crassostrea gigas. (United States)

    Song, Xiaorui; Wang, Hao; Chen, Hao; Sun, Mingzhe; Liang, Zhongxiu; Wang, Lingling; Song, Linsheng


    Hemocytes are the effective immunocytes in bivalves, which have been reported to be derived from stem-like cells in gill epithelium of oyster. In the present work, a conserved haematopoietic transcription factor Tal-1/Scl (Stem Cell Leukemia) was identified in Pacific oyster (Cg-SCL), and it was evolutionarily close to the orthologs in deuterostomes. Cg-SCL was highly distributed in the hemocytes as well as gill and mantle. The hemocyte specific genes Integrin, EcSOD and haematopoietic transcription factors GATA3, C-Myb, c-kit, were down-regulated when Cg-SCL was interfered by dsRNA. During the larval developmental stages, the mRNA transcripts of Cg-SCL gradually increased after fertilization and peaked at early trochophore larvae stage (10 hpf, hours post fertilization), then sharply decreased in late trochophore larvae stage (15 hpf) before resuming in umbo larvae (120 hpf). Whole-mount immunofluorescence assay further revealed that the immunoreactivity of Cg-SCL appeared in blastula larvae with two approximate symmetric spots, and this expression pattern lasted in gastrula larvae. By trochophore, the immunoreactivity formed a ring around the dorsal region and then separated into two remarkable spots at the dorsal side in D-veliger larvae. After bacterial challenge, the mRNA expression levels of Cg-SCL were significantly up-regulated in the D-veliger and umbo larvae, indicating the available hematopoietic regulation in oyster larvae. These results demonstrated that Cg-SCL could be used as haematopoietic specific marker to trace potential developmental events of hematopoiesis during ontogenesis of oyster, which occurred early in blastula stage and maintained until D-veliger larvae. PMID:26915307

  4. Regulation of inhibin βB-subunit mRNA expression in rat Sertoli cells: Consequences for the production of bioactive and immunoreactive inhibin

    NARCIS (Netherlands)

    K. Klaij (Kevin); M.A. Timmerman (Marianna); L.J. Blok (Leen); J.A. Grootegoed (Anton); F.H. de Jong (Frank)


    markdownabstractAbstract In Sertoli cells from 21-day-old rats, the expression of the mRNA encoding the α-subunit of inhibin, and the production of immunoreactive inhibin are stimulated by follicle-stimulating hormone (FSH). In contrast, the amount of βB-subunit mRNA is not increased after FSH tre

  5. Biosynthesis and release of thyrotropin-releasing hormone immunoreactivity in rat pancreatic islets in organ culture. Effects of age, glucose, and streptozotocin

    DEFF Research Database (Denmark)

    Dolva, L O; Welinder, B S; Hanssen, K F;


    Thyrotropin-releasing hormone immunoreactivity (TRH-IR) was measured in isolated islets and in medium from rat pancreatic islets maintained in organ culture. TRH-IR in methanol extracts of both islets and culture medium was eluted in the same position as synthetic TRH by ion-exchange and gel...


    NARCIS (Netherlands)



    Changes in neocortical immunoreactivity (ir) for muscarinic acetylcholine receptors (mAChRs), protein kinase C gamma (PKC gamma), microtubule-associated protein 2 (MAP-2), and the calcium-binding protein parvalbumin (PARV) induced by the performance of a one-trial passive shock avoidance (PSA) task

  7. Exposure to chronic psychosocial stress and corticosterone in the rat : Effects on spatial discrimination learning and hippocampal protein kinase C gamma immunoreactivity

    NARCIS (Netherlands)

    Krugers, HJ; Douma, BRK; Bohus, B; Korf, J; Luiten, PGM; Krugers, Harm J.


    Previous reports have demonstrated a striking increase of the immunoreactivity of the gamma-isoform of protein kinase C (PKC gamma-ir) in Ammon's horn and dentate gyrus (DC) of rodent hippocampus after training in a spatial orientation task. In the present study, we investigated how 8 days of psycho

  8. Neurochemical Phenotype of Reelin Immunoreactive Cells in the Piriform Cortex Layer II (United States)

    Carceller, Hector; Rovira-Esteban, Laura; Nacher, Juan; Castrén, Eero; Guirado, Ramon


    Reelin, a glycoprotein expressed by Cajal-Retzius neurons throughout the marginal layer of developing neocortex, has been extensively shown to play an important role during brain development, guiding neuronal migration and detachment from radial glia. During the adult life, however, many studies have associated Reelin expression to enhanced neuronal plasticity. Although its mechanism of action in the adult brain remains mostly unknown, Reelin is expressed mainly by a subset of mature interneurons. Here, we confirm the described phenotype of this subpopulation in the adult neocortex. We show that these mature interneurons, although being in close proximity, lack polysialylated neural cell adhesion molecule (PSA-NCAM) expression, a molecule expressed by a subpopulation of mature interneurons, related to brain development and involved in neuronal plasticity of the adult brain as well. However, in the layer II of Piriform cortex there is a high density of cells expressing Reelin whose neurochemical phenotype and connectivity has not been described before. Interestingly, in close proximity to these Reelin expressing cells there is a numerous subpopulation of immature neurons expressing PSA-NCAM and doublecortin (DCX) in this layer of the Piriform cortex. Here, we show that Reelin cells express the neuronal marker Neuronal Nuclei (NeuN), but however the majority of neurons lack markers of mature excitatory or inhibitory neurons. A detail analysis of its morphology indicates these that some of these cells might correspond to semilunar neurons. Interestingly, we found that the majority of these cells express T-box brain 1 (TBR-1) a transcription factor found not only in post-mitotic neurons that differentiate to glutamatergic excitatory neurons but also in Cajal-Retzius cells. We suggest that the function of these Reelin expressing cells might be similar to that of the Cajal-Retzius cells during development, having a role in the maintenance of the immature phenotype of the

  9. Immunocytochemical localization of glutamic acid decarboxylase (GAD) and substance P in neural areas mediating motion-induced emesis: Effects of vagal stimulation on GAD immunoreactivity (United States)

    Damelio, F.; Gibbs, M. A.; Mehler, W. R.; Daunton, Nancy G.; Fox, Robert A.


    Immunocytochemical methods were employed to localize the neurotransmitter amino acid gamma-aminobutyric acid (GABA) by means of its biosynthetic enzyme glutamic acid decarboxylase (GAD) and the neuropeptide substance P in the area postrema (AP), area subpostrema (ASP), nucleus of the tractus solitarius (NTS), and gelatinous nucleus (GEL). In addition, electrical stimulation was applied to the night vagus nerve at the cervical level to assess the effects on GAD-immunoreactivity (GAR-IR). GAD-IR terminals and fibers were observed in the AP, ASP, NTS, and GEL. They showed pronounced density at the level of the ASP and gradual decrease towards the solitary complex. Nerve cells were not labelled in our preparations. Ultrastructural studies showed symmetric or asymmetric synaptic contracts between labelled terminals and non-immunoreactive dendrites, axons, or neurons. Some of the labelled terminals contained both clear- and dense-core vesicles. Our preliminary findings, after electrical stimulation of the vagus nerve, revealed a bilateral decrease of GAD-IR that was particularly evident at the level of the ASP. SP-immunoreactive (SP-IR) terminals and fibers showed varying densities in the AP, ASP, NTS, and GEL. In our preparations, the lateral sub-division of the NTS showed the greatest accumulation. The ASP showed medium density of immunoreactive varicosities and terminals and the AP and GEL displayed scattered varicose axon terminals. The electron microscopy revealed that all immunoreactive terminals contained clear-core vesicles which make symmetric or asymmetric synaptic contact with unlabelled dendrites. It is suggested that the GABAergic terminals might correspond to vagal afferent projections and that GAD/GABA and substance P might be co-localized in the same terminal allowing the possibility of a regulated release of the transmitters in relation to demands.

  10. Relationships among parvalbumin-immunoreactive neuron density, phase-locked gamma oscillations, and autistic/schizophrenic symptoms in PDGFR-β knock-out and control mice.

    Directory of Open Access Journals (Sweden)

    Tomoya Nakamura

    Full Text Available Cognitive deficits and negative symptoms are important therapeutic targets for schizophrenia and autism disorders. Although reduction of phase-locked gamma oscillation has been suggested to be a result of reduced parvalbumin-immunoreactive (putatively, GABAergic neurons, no direct correlations between these have been established in these disorders. In the present study, we investigated such relationships during pharmacological treatment with a newly synthesized drug, T-817MA, which displays neuroprotective and neurotrophic effects. In this study, we used platelet-derived growth factor receptor-β gene knockout (PDGFR-β KO mice as an animal model of schizophrenia and autism. These mutant mice display a reduction in social behaviors; deficits in prepulse inhibition (PPI; reduced levels of parvalbumin-immunoreactive neurons in the medical prefrontal cortex, hippocampus, amygdala, and superior colliculus; and a deficit in of auditory phase-locked gamma oscillations. We found that oral administration of T-817MA ameliorated all these symptoms in the PDGFR-β KO mice. Furthermore, phase-locked gamma oscillations were significantly correlated with the density of parvalbumin-immunoreactive neurons, which was, in turn, correlated with PPI and behavioral parameters. These findings suggest that recovery of parvalbumin-immunoreactive neurons by pharmacological intervention relieved the reduction of phase-locked gamma oscillations and, consequently, ameliorated PPI and social behavioral deficits. Thus, our findings suggest that phase-locked gamma oscillations could be a useful physiological biomarker for abnormality of parvalbumin-immunoreactive neurons that may induce cognitive deficits and negative symptoms of schizophrenia and autism, as well as of effective pharmacological interventions in both humans and experimental animals.