ABSTRACT
Physical exercise exerts favourable effects on brain health and quality of life of the elderly; some of these positive health effects are induced by the modulation of microbiota composition. We therefore conducted a randomised, double blind, placebo-controlled trial that assessed whether a combination of Bifidobacterium spp. supplementation and moderate resistance training improved the cognitive function and other health-related parameters in healthy elderly subjects. Over a 12-week period, 38 participants (66-78 years) underwent resistance training and were assigned to the probiotic Bifidobacterium supplementation (n=20; 1.25×1010 cfu each of Bifidobacterium longum subsp. longum BB536, B. longum subsp. infantis M-63, Bifidobacterium breve M-16V and B. breve B-3) or the placebo (n=18) group. At baseline and at 12 weeks, we assessed the cognitive function, using the Japanese version of the Montreal Cognitive Assessment instrument (MoCA-J); modified flanker task scores; depression-anxiety scores; body composition; and bowel habits. At 12 weeks, the MoCA-J scores showed a significant increase in both the groups, while the flanker task scores of the probiotic group increased more significantly than those of the placebo group (0.35±0.9 vs -0.29±1.1, P=0.056). Only the probiotic group showed a significant decrease in the depression-anxiety scores (5.2±6.3 to 3.4±5.5, P=0.012) and body mass index (24.0±2.8 to 23.5±2.8 kg/m2, P<0.001), with a significant increase in the defecation frequency (5.3±2.3 to 6.4±2.3 times/5 days, P=0.023) at 12 weeks. Thus, in healthy elderly subjects, combined probiotic bifidobacteria supplementation and moderate resistance training may improve the mental condition, body weight and bowel movement frequency.
Subject(s)
Bifidobacterium/growth & development , Dietary Supplements , Healthy Volunteers , Probiotics/administration & dosage , Resistance Training , Aged , Animals , Body Composition , Cognition , Defecation , Double-Blind Method , Female , Humans , Male , Placebos/administration & dosage , Treatment OutcomeABSTRACT
RESUMO Tivemos como objetivo avaliar o efeito da infusão de Cunila microcephala Benth sobre a atividade da enzima acetilcolinesterase (AChE) e marcadores de estresse oxidativo em eritrócitos de agricultores. Foram utilizadas amostras provenientes de 16 trabalhadores rurais expostos a pesticidas agrícolas pelo período mínimo de 5 anos e um grupo controle constituído de 16 indivíduos não expostos a agrotóxicos. As hemácias dos agricultores e o grupo A foram expostos “in vitro” à solução salina (NaCl 0,9%). Os demais grupos foram expostos à infusão de poejo nas concentrações de 0; 5; 10; 25 e 50 g/L (Grupos B; C; D e E, respectivamente). Em seguida, foram realizadas as determinações da atividade da AChE e dos níveis de substâncias reativas ao ácido tiobarbitúrico (TBARS), proteínas carboniladas (PCs) e glutationa reduzida (GSH). Os resultados mostram que a infusão de poejo 50g/L, aumenta a atividade da enzima AChE e os níveis de GSH. Contudo, os níveis de TBARS e PCs diminuíram após o tratamento com a infusão de poejo 25 e 50 g/L. A infusão de poejo, na concentração de 50 g/L, é capaz de reverter, “in vitro” a inibição da atividade da AChE que ocorre pela exposição a pesticidas, e ainda demonstra um importante potencial antioxidante, tendo em vista que diminuiu danos lipídicos e proteicos e ainda, estimulou a produção do principal antioxidante não enzimático endógeno.
ABSTRACT Evaluating the effect of infusion Cunila microcephala Benth on acetylcholinesterase activity (AChE) enzyme and on biomarkers of oxidative stress in farmers erythrocytes. We used samples from 16 rural workers exposed to pesticides for a minimum of five years, and a control group composed of 16 individuals not exposed to pesticides. The erythrocytes of farmers and from group A were exposed “in vitro” the saline solution (NaCl 0,9%). The other groups were exposed to the infusion of “poejo” at concentrations of 0; 5; 10; 25 and 50 g/L (Groups B, C, D and E, respectively). Then, it was realized the analitical determinations of AChE activity and TBARS, PCs and GSH levels. The results showed that “poejo” infusion 50g/L, increased the AChE activity and GSH levels. However, the TBARS e PCs levels decreased after the treatment with “poejo” infusion 25 e 50 g/L. The “poejo” infusion 50 g/L is able to revert “in vitro” the inhibition of AChE activity that occurs by exposure to pesticides and also demonstrates an important antioxidant potential, given that decreased lipid and protein damage and also it stimulated the production of the main non-enzymatic antioxidant endogenous.
Subject(s)
Humans , Acetylcholinesterase/pharmacology , In Vitro Techniques/instrumentation , Biomarkers/analysis , Oxidative Stress , Lamiaceae/classification , Pesticides , Erythrocytes/classification , Farmers/classificationSubject(s)
Internationality/history , Medicine, East Asian Traditional/history , Obstetrics/history , Female , Humans , PregnancyABSTRACT
Urtica dioica and Convolvulus arvensis are the main host plants of Hyalesthes obsoletus and play an important role in the epidemiology of Bois noir of grapevines. The earliest survey, which was carried out to compare the phenology of nymphal instars on U. dioica and C. arvensis, had highlighted some problems in the identification of the instars. Therefore, the correct identification of nymphs to species and instar level became a preliminary aim of this research. Adults and nymphs attributable to H. obsoletus were collected during 2008-2010 in three flatland vineyard habitats of northern Italy on U. dioica, C. arvensis and Artemisia verlotorum. Nymphs and morphologically identified adults of H. obsoletus were submitted to molecular identification. Morphometric and morphological studies were carried out on nymphs collected in the field or obtained in laboratory rearings. Molecular methods not only confirmed the identity of adults, but also allowed the assignment of the nymphs to this species. Morphometric and morphological characteristics (e.g. body and head-thoracic lengths, number of thoracic pits) showed the existence of five nymphal instars. Morphometric differences between newly hatched and older first-instar nymphs were observed. A key to distinguish the five instars was proposed. Evident differences between H. obsoletus nymphs studied here and elsewhere were identified. According to differences in adult-flight period, an earlier phenology of nymphs on C. arvensis than on U. dioica was observed. In particular, the typical overwintering instar was the second on U. dioica and the third on C. arvensis.
Subject(s)
Hemiptera/anatomy & histology , Hemiptera/classification , Animals , Artemisia/physiology , Convolvulus/physiology , Electron Transport Complex IV/genetics , Female , Hemiptera/genetics , Hemiptera/physiology , Herbivory , Insect Control , Insect Proteins/genetics , Italy , Male , Mitochondrial Proteins/genetics , Mitochondrial Proteins/physiology , Nymph/anatomy & histology , Nymph/classification , Nymph/genetics , Nymph/physiology , Polymerase Chain Reaction , Urtica dioica/physiologyABSTRACT
Traditional Chinese herbal medicines are frequently prescribed in pharmacotherapy in Japan. In the present study, we evaluated the possible interaction of several herbal extracts including Rhei Rhizoma extract with cytochrome P450 (CYP) 3A and efflux transporters such as P-glycoprotein and multidrug resistance-associated protein (MRP) 2. Rhei Rhizoma extract (100 microg/ml) significantly suppressed the CYP3A-mediated 6beta-hydroxylation of testosterone in hepatic microsomes, and increased the extent of bioavailability of midazolam, a typical CYP3A substrate, in rats. Also, Rhei Rhizoma extract (300 microg/ml) significantly suppressed P-glycoprotein-mediated efflux transport of rhodamine 123 (Rho123) in rat everted intestine. In an in-vivo study, Rhei Rhizoma extract added to intestinal perfusate at a concentration of 300 microg/ml significantly suppressed the intestinal exsorption of Rho123, though it exerted no effect on the biliary excretion of Rho123. Furthermore, the in-vitro and in-vivo MRP2-mediated intestinal efflux of 2,4-dinitrophenyl-S-glutathione was significantly suppressed by Rhei Rhizoma extract (1000 microg/ml). In conclusion, Rhei Rhizoma extract, which is taken orally at doses of 0.5-1 g each or 1-3 g daily in clinical practice, may cause pharmacokinetic herb-drug interactions in the process of the intestinal and/or hepatic CYP3A-mediated drug metabolism and P-glycoprotein- and/or MRP2-mediated efflux transport in the intestine.
Subject(s)
Carrier Proteins/metabolism , Cytochrome P-450 CYP3A/metabolism , Rheum/chemistry , ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , ATP-Binding Cassette Transporters/metabolism , Animals , Cyclosporine/pharmacology , Dinitrochlorobenzene/metabolism , GABA Modulators/pharmacokinetics , Glutathione/analogs & derivatives , Glutathione/metabolism , Immunosuppressive Agents/pharmacology , Indicators and Reagents , Intestinal Mucosa/metabolism , Intestines/drug effects , Liver/drug effects , Liver/metabolism , Male , Microsomes, Liver/drug effects , Microsomes, Liver/enzymology , Midazolam/pharmacokinetics , Plant Extracts/pharmacology , Rats , Rats, Sprague-Dawley , Rhodamine 123ABSTRACT
The death of honey bees, Apis mellifera L., and the consequent colony collapse disorder causes major losses in agriculture and plant pollination worldwide. The phenomenon showed increasing rates in the past years, although its causes are still awaiting a clear answer. Although neonicotinoid systemic insecticides used for seed coating of agricultural crops were suspected as possible reason, studies so far have not shown the existence of unquestionable sources capable of delivering directly intoxicating doses in the fields. Guttation is a natural plant phenomenon causing the excretion of xylem fluid at leaf margins. Here, we show that leaf guttation drops of all the corn plants germinated from neonicotinoid-coated seeds contained amounts of insecticide constantly higher than 10 mg/l, with maxima up to 100 mg/l for thiamethoxam and clothianidin, and up to 200 mg/l for imidacloprid. The concentration of neonicotinoids in guttation drops can be near those of active ingredients commonly applied in field sprays for pest control, or even higher. When bees consume guttation drops, collected from plants grown from neonicotinoid-coated seeds, they encounter death within few minutes.
Subject(s)
Bees/drug effects , Insecticides/toxicity , Abdomen/physiology , Animals , Bees/physiology , Colony Collapse , Imidazoles/toxicity , Neonicotinoids , Nitro Compounds/toxicity , Plant Extracts/pharmacology , Plant Leaves/physiology , Pollination/physiology , Seedlings/physiology , Seeds/physiology , Wings, Animal/drug effects , Wings, Animal/physiology , Zea mays/physiologyABSTRACT
The involvement of oxidative stress has been suggested as a mechanism for neurotoxicity caused by methylmercury (MeHg), but the mechanism for MeHg selective toxicity in the central nervous system is still unclear. In this research, to clarify the mechanism of selective neurotoxicity caused by MeHg, the oxygen consumption levels, the reactive oxygen species (ROS) production rates and several antioxidant levels in mitochondria were compared among the cerebrum, cerebellum and liver of male Wistar rats. In addition, the alterations of these indexes were examined in MeHg-intoxicated rats (oral administration of 10 mg/kg day, for 5 days). Although the cerebrum and cerebellum in intact rats showed higher mitochondrial oxygen consumption levels and ROS production rates than the liver, glutathione peroxidase (GPX) and superoxide dismutase (SOD) activities were much lower in the cerebrum and cerebellum than in the liver. Especially, the cerebellum showed the highest oxygen consumption and ROS production rate and the lowest mitochondrial glutathione (GSH) levels among the tissues examined. In the MeHg-treated rats, decrease in the oxygen consumption and increase in the ROS generation were found only in the cerebellum mitochondria, despite a lower Hg accumulation in the mitochondrial fraction compared to the liver. Since MeHg treatment produced an enhancement of ROS generation in cerebellum mitochondria supplemented with succinate substrates, MeHg-induced oxidative stress might affect the complex II-III mediated pathway in the electron transfer chain in the cerebellum mitochondria. Our study suggested that inborn factors, high production system activity and low defense system activity of ROS in the brain, would relate to the high susceptibility of the central nervous system to MeHg toxicity.
Subject(s)
Cerebellum/drug effects , Hydrogen Peroxide/metabolism , Methylmercury Compounds/toxicity , Mitochondria/drug effects , Superoxides/metabolism , Animals , Cerebellum/metabolism , Cerebrum/drug effects , Cerebrum/metabolism , Glutathione Peroxidase/metabolism , Liver/drug effects , Liver/metabolism , Male , Mitochondria/metabolism , Oxygen Consumption/drug effects , Rats , Rats, Wistar , Superoxide Dismutase/metabolismABSTRACT
Previously isolated RanBPM, a Ran-binding protein in the microtubule-organizing center, which had been thought to play a role in Ran-stimulated microtubule assembly, turned out to be a truncated protein. To clarify the function of RanBPM, we cloned the full-sized RanBPM cDNA that encodes a 90 kDa protein, compared to the previously isolated cDNA that encoded a 55 kDa protein. The newly cloned 5' coding region contains a great number of cytidine and guanidine nucleotides, like the CpG island. Thus, full-sized RanBPM cDNA encodes a long stretch of proline and glutamine residues in the N-terminal region. It comprises a protein complex of more than 670 kDa. Ran was detected in this complex when RanBPM and Ran were both ectopically expressed. New antibodies to RanBPM were prepared against three different regions of RanBPM. All of them detected a 90 kDa protein that is predominantly localized both in the nucleus and in the cytoplasmic region surrounding the centrosome, but none of them stained the centrosome. In this context, our previous notion that RanBPM is a centrosomal protein should be discarded. RanBPM is well conserved in the animal kingdom. It may play an important role in uncovering Ran-dependent nuclear events.
Subject(s)
DNA, Complementary/genetics , ran GTP-Binding Protein/genetics , Adaptor Proteins, Signal Transducing , Amino Acid Sequence , Animals , CHO Cells , COS Cells , Cell Line , Centrosome/chemistry , Cricetinae , Cytoskeletal Proteins , DNA, Complementary/chemistry , Glutamine/genetics , HeLa Cells , Humans , Immunoblotting , KB Cells , Molecular Sequence Data , Nuclear Proteins , Proline/genetics , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology, Amino Acid , ran GTP-Binding Protein/analysisABSTRACT
Eight patients with large vestibular aqueduct (LVA) syndrome and acute sensorineural hearing loss that had not responded to intravenous treatment were treated with hyperbaric oxygen(HBO2) therapy. This was performed daily, with 15 treatments per course. Pure tone and speech audiometry were performed on all patients. The mean hearing recovery following HBO2 therapy (20.8 +/- 12.1 dB) was significantly higher than after intravenous treatment (2.3 +/- 7.2 dB) (P < 0.01), although treatment duration was not different between the two therapies. Of the eight patients, five showed complete or partial recovery after one course of HBO2 therapy, and their hearing levels have not since changed. The other three patients showed heating recovery during therapy, but treatment had to be repeated for two or more courses because of progressive or fluctuating hearing loss.We consider HBO2 therapy should be used to treat acute hearing deterioration associated with LVA syndrome if patients do not recover their hearing ability following conventional treatment
Subject(s)
Deafness/therapy , Hearing Loss/therapy , Hyperbaric Oxygenation , Vestibular Aqueduct/abnormalities , Adult , Child , Child, Preschool , Deafness/genetics , Female , Hearing Loss/genetics , Humans , Male , Mutation, Missense , Recovery of Function , SyndromeABSTRACT
In September 1996, an outbreak of methicillin-resistant Staphylococcus aureus (MRSA) colonization occurred in the neonatal intensive care unit (NICU) of our hospital. After failing to control the outbreak by conventional methods we implemented an intranasal blanket use programme of mupirocin ointment from the beginning of November 1997. In the programme, patients who had been carrying MRSA received intranasal administration of the ointment three times daily for the first three days and consecutively three times weekly, while newly admitted patients and those who had not been colonized were prophylactically medicated three times weekly. This blanket administration was executed for one month. Methicillin-resistant Staphylococcus aureus colonization became undetectable in all but one intubated inpatient who had already been colonized before the start of the programme, and no new acquisitions occurred until the middle of January 1998, seven weeks after the termination of the blanket use programme. The rate of colonized patients in the unit also decreased. During and after the programme, neither an increase in minimum inhibitory concentration for the antibiotic nor apparent adverse reactions in any of the treated patients were observed. We concluded that this procedure is an effective method of controlling an MRSA outbreak in an NICU when the outbreak cannot be managed with conventional measures.
Subject(s)
Anti-Bacterial Agents/therapeutic use , Carrier State/prevention & control , Cross Infection/prevention & control , Disease Outbreaks/prevention & control , Infection Control/methods , Intensive Care Units, Neonatal , Methicillin Resistance , Mupirocin/therapeutic use , Nasal Mucosa/microbiology , Staphylococcal Infections/prevention & control , Staphylococcus aureus , Administration, Intranasal , Carrier State/epidemiology , Carrier State/microbiology , Cross Infection/epidemiology , Cross Infection/microbiology , DNA Fingerprinting , DNA, Bacterial/analysis , DNA, Bacterial/genetics , Disease Outbreaks/statistics & numerical data , Humans , Infant, Newborn , Microbial Sensitivity Tests , Ointments , Program Evaluation , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology , Staphylococcus aureus/classification , Staphylococcus aureus/genetics , Time Factors , TokyoABSTRACT
Methionine is a protective factor against various types of liver damage, but excessive dietary methionine is hepatotoxic. Because the mechanisms of L-methionine-related hepatotoxicity are poorly understood, the effect of long-term excessive L-methionine intake on the metabolism of iron and antioxidants was studied in rat liver to determine whether oxidative stress is involved. Wistar male rats were fed either an L-methionine-supplemented (16.0 g/kg) diet or a control diet for 1, 3, 6 and 9 mo. The growth rate of L-methionine-supplemented rats was significantly slower than that of controls. Iron, ferritin and thiobarbituric acid-reactive substances (TBARS) levels in the liver were greater in supplemented rats than in controls. Serum iron and transferrin levels were significantly lower in L-methionine-treated rats compared with controls. Serum ferritin did not differ between the two groups. Hepatic glutathione peroxidase activity, catalase activity and total glutathione concentrations were higher in rats fed the L-methionine-supplemented diet at 1 and 3 mo, but not at 6 and 9 mo. These results indicate that long-term consumption of excess L-methionine by rats may affect primarily iron metabolism rather than the antioxidant defense system and, consequently, induce an accumulation of iron.
Subject(s)
Iron/metabolism , Lipid Peroxidation/drug effects , Liver/drug effects , Methionine/pharmacology , Animals , Antioxidants/metabolism , Body Weight/drug effects , Catalase/metabolism , Diet , Dose-Response Relationship, Drug , Ferritins/metabolism , Glutathione Peroxidase/metabolism , Iron/blood , Liver/enzymology , Liver/metabolism , Male , Methionine/administration & dosage , Oxidative Stress , Rats , Rats, Wistar , Thiobarbituric Acid Reactive Substances/metabolismABSTRACT
Recent genetic studies of tumorigenesis have strongly suggested an existence of tumor suppressor gene(s) on murine chromosome 12 and human chromosome 14q32. We previously described that putative tumor suppressor gene(s) might reside between D12Mit53 and D12Mit233. We analyzed three genes, Tcl1, Yy1 and Tnfalphaip2, which had been mapped around the region, as the candidates in radiation lymphomagenesis of (BALB/c x MSM/Ms)F1 hybrid mice. The locus order and distances of the three genes and microsatellite loci were estimated as follows: [centromere] - Tcl1-(> or =0.085 cM)-D12Mit50-(0.085 cM)D12Mit132-(1.96 cM)D12Mit122-(0.085 cM)D12Mit53-(1.37 cM)-[D12Mit233,D12Mit279,Yy1]-(0.085 cM)-D12Mit181-(> or =0.17 cM)-Tnfalphaip2 - [telomere]. Allele losses at Tcl1, Yy1 genes and D12Mit233 were observed in 94(45%), 143(68%) and 147(70%) of 210 lymphomas, respectively. In semi-quantitative analysis of Yy1 mRNA levels by RT-PCR, kinetics of the yield of the Yy1-cDNA-specific PCR products showed almost the same profiles among thymic lymphomas with allelic loss at Yy1, lymphomas with both alleles retained and normal thymus. These results suggest that Tcl1, Yy1 and Tnfalphaip2 genes are not predominantly involved in radiation lymphomagenesis of mice. In further analysis of the common allelic loss region, we found new loci, Y152pR1 and Y184pR2, from YACs which located in the hot region between D12Mit53 and D12Mit233, and the highest frequency of allelic loss (71%) was observed at the Y184pR2 locus. The LOH patterns of individual lymphomas suggest that putative tumor suppressor gene(s) lies between Y152pR1 and Y184pR2.
Subject(s)
Genes, Tumor Suppressor/genetics , Loss of Heterozygosity , Lymphoma/genetics , Neoplasms, Radiation-Induced/genetics , Proto-Oncogene Proteins , Animals , Chromosome Mapping , DNA-Binding Proteins/genetics , Erythroid-Specific DNA-Binding Factors , Female , Haplotypes , Lymphoma/etiology , Male , Mice , Mice, Inbred BALB C , Mice, Inbred Strains , Microsatellite Repeats/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Recombination, Genetic/genetics , Testis/metabolism , Thymus Gland/metabolism , Thymus Gland/pathology , Thymus Gland/radiation effects , Transcription Factors/genetics , Tumor Necrosis Factor-alpha/genetics , YY1 Transcription FactorABSTRACT
Synapse replacement after brain injury has been widely documented by anatomical studies in various parts of both the developing and adult nervous system. However, the molecular events that define the specificity of the empirically derived rules of reactive synaptogenesis in different regions of the adult brain remain unclear. In this study we examined the differential regulation of the lesion-induced response of the two growth-associated proteins, superior cervical ganglia-10 and growth-associated protein-43, after unilateral cortex ablation, and determined a hierarchical order for the lesion response from remaining afferent projection neurons originating from the contralateral cortex, ipsilateral thalamus and substantia nigra. We report that in response to unilateral cortex ablation both messenger RNA, by northern blot, and protein, by western blot, for superior cervical ganglia-10 but not growth-associated protein-43 was increased in the homologous area of the contralateral cortex but not the ipsilateral thalamus or substantia nigra. In addition, the specificity of the superior cervical ganglia-10 response, assessed by combined in situ hybridization and retrograde FluoroGold labeling of striatal afferent neurons, found that superior cervical ganglia-10 messenger RNA was increased prominently in layer V pyramidal neurons of the contralateral corticostriatal pathway but was unchanged in afferent projection neurons from the thalamus and substantia nigra. Furthermore, the increase in both superior cervical ganglia-10 messenger RNA and protein seen at three days postlesion in contralateral corticostriatal neurons coincides in time with the initiation of neurite outgrowth in the deafferented striatum by contralateral corticostriatal axons described in our previous ultrastructural study. However, if cortical input to the striatum was removed bilaterally the lesion-induced response for superior cervical ganglia-10 messenger RNA shifted secondarily to thalamostriatal neurons in the ipsilateral thalamus. These data provide evidence that superior cervical ganglia-10 and growth-associated protein-43 are differentially regulated in neurons of the contralateral corticostriatal pathway in response to unilateral cortex ablation and suggests that superior cervical ganglia-10 plays a role in the regulation of neurite outgrowth in the adult striatum after brain injury. However, the specific role that superior cervical ganglia-10 may play in reactive synaptogenesis remains unclear. In addition, our data suggest that a hierarchical order exists for the reinnervation of deafferented striatal neurons after unilateral cortex ablation with preference given to homologous axons from the contralateral cortex.
Subject(s)
Brain Injuries/metabolism , Cerebral Cortex/physiology , Dominance, Cerebral/physiology , GAP-43 Protein/metabolism , Nerve Growth Factors/metabolism , Stilbamidines , Animals , Blotting, Northern , Carrier Proteins , Cerebral Cortex/cytology , Cerebral Cortex/metabolism , Fluorescent Dyes/pharmacokinetics , GAP-43 Protein/genetics , In Situ Hybridization , Male , Membrane Proteins , Microtubule Proteins , Nerve Growth Factors/genetics , Neurons/metabolism , RNA, Messenger/metabolism , Rats , Rats, Inbred F344 , Substantia Nigra/cytology , Substantia Nigra/metabolism , Thalamus/cytology , Thalamus/metabolism , Time Factors , Tissue Distribution/physiologyABSTRACT
Shc has been implicated in a variety of growth factor- and cytokine receptor-signaling through its specific binding to phosphotyrosine residues of the activated receptors. In neuronal cells, such as PC12, Shc has been shown to be involved in Ras-dependent MAP kinase activation following Trk receptor stimulation with NGF. While the ubiquitous role of Shc as an adaptor molecule in signal transduction is increasing in both neuronal and non-neuronal cells and tissues, the expression level of Shc is surprisingly low in the brain. We demonstrated here the isolation of a neural-specific member of the Shc family. This novel protein, named N-Shc (neuronal Shc), contains two potential phosphotyrosine-binding domains, PTB and SH2, and is expressed exclusively in the brain; whereas Shc is present in all other non-neuronal tissues. As in Shc, N-Shc can bind activated EGF receptor, become tyrosine phosphorylated, and form a complex with Grb2 adapter protein following EGF stimulation. Furthermore, N-Shc can bind activated TrkB receptor following the stimulation with brain-derived neurotrophic factor (BDNF), which is the most abundant neurotrophin in the brain. These data suggest that N-Shc, rather than Shc, mediates neurotrophin and other neuronal signalings in the central nervous system.
Subject(s)
Adaptor Proteins, Signal Transducing , Brain-Derived Neurotrophic Factor/physiology , Calcium-Calmodulin-Dependent Protein Kinases/physiology , Receptor Protein-Tyrosine Kinases/physiology , Receptors, Nerve Growth Factor/physiology , Signal Transduction/physiology , ras Proteins/physiology , src Homology Domains/physiology , 3T3 Cells , Amino Acid Sequence , Animals , Base Sequence , Brain/physiology , Brain-Derived Neurotrophic Factor/metabolism , COS Cells , Calcium-Calmodulin-Dependent Protein Kinases/metabolism , Cell Differentiation/physiology , Cell Division/physiology , Cell Survival/physiology , DNA, Complementary/genetics , DNA, Complementary/isolation & purification , Epidermal Growth Factor/metabolism , Epidermal Growth Factor/physiology , ErbB Receptors/metabolism , ErbB Receptors/physiology , GRB2 Adaptor Protein , Humans , Mice , Molecular Sequence Data , Neurons/cytology , Neurons/metabolism , Neurons/physiology , Phosphorylation , Proteins/metabolism , Proteins/physiology , Receptor Protein-Tyrosine Kinases/metabolism , Receptor, trkB , Receptors, Nerve Growth Factor/metabolism , Sensitivity and Specificity , Sequence Homology, Amino Acid , Tyrosine/metabolism , ras Proteins/metabolismABSTRACT
Cardiohemodynamic effects of 3'-angeloyloxy-4'-acetoxy-3',4'-dihydroseselin (Pd-Ia) isolated from the Chinese medicinal plant Peucedanum praeruptorum Dunn were compared with those of diltiazem (Dil) in anesthetized open-chest dogs. Pd-Ia 3 mg.kg-1 increased coronary blood flow from 25 +/- 11 to 58 +/- 16 ml.min-1 (n = 7, P < 0.01) and decreased mean aortic pressure (MAP) from 13 +/- 2 to 8 +/- 1 kPa (P < 0.01), rate pressure product (RPP) from 1.9 +/- 0.5 to 1.3 +/- 0.3 MPa.bpm (P < 0.05), +dp/dtmax from 246 +/- 56 to 160 +/- 36 kPa.s-1 (P < 0.01) and systemic vascular resistance from 19 +/- 4 to 12 +/- 7 Pa.ml-1.min-1 (P < 0.05), together with an increase in HR. Dil showed effects similar to those of Pd-Ia except for a marked decrease in HR. The effects of Pd-Ia on MAP and RPP were approximately one-tenth as potent as those of Dil. The results demonstrated that Pd-Ia was a Ca2+ channel blocker.
Subject(s)
Blood Pressure/drug effects , Calcium Channel Blockers/pharmacology , Coronary Circulation/drug effects , Coumarins/pharmacology , Animals , Diltiazem/pharmacology , Dogs , Dose-Response Relationship, Drug , Drugs, Chinese Herbal/chemistry , Female , Heart Rate/drug effects , Male , Vascular Resistance/drug effectsABSTRACT
Expression of mineralocorticoid type I receptor (MR) gene in the rat cochlea was determined using molecular biological techniques. We synthesized complementary DNA (cDNA) from rat cochlear total RNA and then amplified MR cDNA fragments by polymerase chain reaction (PCR). The amplified cDNA fragments were subcloned into an expression vector and the nucleotide sequence was analyzed to confirm the expression of mRNA encoding MR in the cochlea. We then synthesized digoxigenin-labeled riboprobes with this cloned DNA template and examined the localization of MR mRNA in the cochlea by in situ hybridization. The amino acid sequence of MR cDNA expressed in the cochlea was identical to that of the MR first cloned in the rat hippocampus. In situ hybridization showed the expression of MR mRNA in marginal cells of the stria vascularis, suggesting that aldosterone may regulate microhomeostasis of the endolymph, presumably by modulating Na, K-ATPase activity. Intense MR signal was also identified in spiral ganglion cells, the function of which remains to be determined.
Subject(s)
Cochlea/metabolism , Gene Expression Regulation/genetics , RNA, Messenger/metabolism , Receptors, Mineralocorticoid/genetics , Aldosterone/metabolism , Aldosterone/pharmacology , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , DNA Primers/chemistry , DNA, Complementary/biosynthesis , DNA, Complementary/chemistry , Endolymph/drug effects , Endolymph/enzymology , Female , Hippocampus/metabolism , Homeostasis/drug effects , In Situ Hybridization , Kidney Cortex/metabolism , Molecular Sequence Data , Polymerase Chain Reaction , RNA, Messenger/genetics , Rats , Rats, Wistar , Receptors, Mineralocorticoid/metabolism , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid , Sodium-Potassium-Exchanging ATPase/metabolism , Spiral Ganglion/metabolism , Spiral Ganglion/ultrastructure , Stria Vascularis/cytology , Stria Vascularis/metabolismABSTRACT
A pyranocoumarin compound, 3'-angeloyloxy-4'-acetoxy-3',4'-dihydroseselin (Pd-Ia), isolated from Bai-Hua Qian-Hu, a Chinese medicine, is known to possess a Ca2+ channel antagonist action. We examined the effect of Pd-Ia on cardiohemodynamics and regional myocardial dysfunction after transient ischemia in anesthetized open-chest dogs. I.v. injections of Pd-Ia (0.1-3.0 mg/kg, n = 7) significantly and dose dependently increased coronary blood flow and decreased mean aortic pressure, maximal rate of rise in left ventricular pressure, rate pressure product and systemic vascular resistance, together with a slight increase in heart rate. Diltiazem (0.03-1.00 mg/kg, n = 6) showed effects on the cardiohemodynamics similar to those of Pd-Ia, except for a marked decrease in heart rate. The effects of Pd-Ia on mean aortic pressure and rate pressure product were approximately 10 times less potent than those of diltiazem. In the second experiment, the effect of a 30-min infusion of Pd-Ia (0.15 mg/kg/min) or vehicle on regional myocardial dysfunction was examined in 16 anesthetized dogs subjected to a 15-min occlusion of the left anterior descending coronary artery followed by a 3-h reperfusion. Infusion of Pd-Ia was started 15 min before coronary occlusion. The vehicle caused a decrease in percent segment shortening throughout the reperfusion period (n = 8). However, Pd-Ia at the dose that produced no significant changes in cardiohemodynamics resulted in a marked improvement in percent segment shortening of the ischemic and reperfused myocardium (n = 8, P < 0.05 vs. control group).(ABSTRACT TRUNCATED AT 250 WORDS)
Subject(s)
Coumarins/pharmacology , Diltiazem/pharmacology , Drugs, Chinese Herbal/therapeutic use , Myocardial Ischemia/drug therapy , Animals , Calcium Channel Blockers/pharmacology , Coronary Circulation/drug effects , Diltiazem/administration & dosage , Disease Models, Animal , Dogs , Dose-Response Relationship, Drug , Female , Hemodynamics/drug effects , Injections, Intravenous , Male , Vascular Resistance/drug effectsABSTRACT
SP iso-response functions in 9 patients with Menière's disease were examined for differences in the properties between +SP and -SP. The iso-response functions of +SP and -SP exhibited a large difference. The iso-response function for +SP was centered at high frequencies (around 9 kHz in most cases) with fairly steep slopes on both sides towards both higher and lower frequencies, whereas -SP showed a low-pass function. The tendency was found for patients with normal hearing at high frequencies to have a larger Q10dB than patients with a hearing loss. The difference in properties between +SP and -SP suggests the possibility that +SP may provide different information on the pathophysiology of Ménière's disease from that obtained from -SP.
Subject(s)
Audiometry, Evoked Response , Meniere Disease/diagnosis , Acoustic Stimulation , Adult , Audiometry, Pure-Tone , Auditory Threshold , Ear, Inner/physiopathology , Electric Stimulation , Electrodes, Implanted , Female , Humans , Male , Meniere Disease/physiopathology , Middle Aged , Tympanic MembraneABSTRACT
The kindling response of the massa intermedia (MI) was assessed in rats. Clinical manifestation of the MI kindling was generally similar to that of limbic kindling, and positive transfer to the amygdala (AM) was obtained following MI kindling. However, MI kindling showed (1) a relatively high after discharge threshold which sometimes increased during the course of kindling, (2) a seizure stage instability with frequent regression to earlier stages, (3) a failure to establish a generalized seizure triggering threshold with an 'all-or-none' property, and (4) a generalized tonic-clonic seizure, which was quite different from a kindled limbic seizure, during early phase of kindling. Furthermore, the MI stimulation caused violent beating movement of the forelimbs, jumping, or running regardless of presence/absence of afterdischarge. The findings suggest that mechanisms other than a simple activation of limbic structures are involved in the process of MI kindling.
Subject(s)
Kindling, Neurologic/physiology , Thalamus/physiopathology , Animals , Electric Stimulation , Limbic System/physiopathology , Male , RatsABSTRACT
Recent studies have demonstrated that intramuscular administration of thyrotropin-releasing hormone (TRH) or its analogue improves various clinical aspects of intractable epilepsy such as Lennox-Gastaut syndrome, West syndrome, and myoclonus epilepsy. Other clinical studies reported efficient property of intravenous TRH against status epilepticus. However, it is also true that intravenous TRH produces epileptic seizures in patients with epilepsy or organic brain damage. Thus, the utility of intravenous TRH for the treatment of status epilepticus seems to be equivocal. To further explore the problem in this regard, we examined the effect of TRH on limbic status epilepticus in rats. Thirty-eight male Wistar rats weighing 180-220g were used. Status epilepticus was induced by intracerebral injection of a combination of 200 micrograms of dibutyryl-cAMP (db-cAMP) and 67.2ng of ethylenediaminetetraacetic acid (EDTA) into the amygdala (AM) through an implanted cannula. 30 min later, TRH or vehicle (distilled water) was administered intravenously (i.v.) or intracerebroventricularly (i.c.v.). Although 3 mg/kg of TRH (n = 9), when injected i.v., did not alter the pattern of electroclinical ictal responses induced by db-cAMP/EDTA, 25 mg/kg (n = 5) and 50 mg/kg (n = 5) of TRH significantly exaggerated EEG and/or behavioral ictal seizures, beginning immediately after the injection and lasting for more than 30 min. With 50 mg/kg of TRH, the exaggerated seizure patterns were followed by marked suppression of electroclinical seizures. 50 micrograms of i.c.v. TRH (n = 5), like higher doses of i.v. TRH, caused a slight, but not a significant, build up of electroclinical ictal seizures, beginning immediately after the injection and lasting for about 30 min.(ABSTRACT TRUNCATED AT 250 WORDS)