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51.
Giannaccini G Betti L Pirone A Palego L Fabiani O Fabbrini L Mascia G Giusti L Macchia M Giusiani M Martini C Lucacchini A 《Neurochemistry international》2007,51(8):496-506
The first effects of 3,4-methylen-dioxy-metamphetamine (MDMA, “ecstasy”), on serotonin 1A (5-HT1A) receptors in rat hippocampus were determined by means of [3H]-8-hydroxy-dipropylamino-tetralin ([3H]-8-OH-DPAT) and 5′guanosine-(γ-[35S]-thio)triphosphate ([35S]-GTPγS) binding as well as inhibition of forskolin (FK)-stimulated adenylyl cyclase (AC) activity. The study was completed by [35S]-GTPγS functional autoradiography experiments carried out in frontal sections of rat brain, including the hippocampal region. Results showed that MDMA was either able to displace [3H]-8-OH-DPAT binding (Ki 500 nM) or to reduce the number of specific sites (Bmax) without affecting Kd. The drug also failed to change the [35S]-GTPγS binding or to inhibit AC velocity, underlying its behavior as a non-competitive 5-HT1A receptor antagonist. Further, MDMA (1 or 100 μM), partially antagonized either [35S]-GTPγS binding stimulation of the agonists 5CT and 8-OH-DPAT or the AC inhibition induced by 5CT and DP-5CT. However, in contrast to binding studies, in AC assays the amphetamine displayed an effect also on EC50, always being less potent than the reference antagonist WAY100,635. In functional autoradiography, MDMA behaved either as a partial 5-HT1A antagonist in limbic areas or, added alone, as an agonist, increasing the coupling signal presumably through 5-HT release from synapses. Interestingly, the selective 5-HT re-uptake inhibitor (SSRI) fluoxetine had no effect on MDMA [35S]-GTPγS binding activation. This latter finding indicates that the amphetamine can release 5-HT via alternative mechanisms to 5-HT transporter binding, probably via membrane synaptic receptors or vesicular transporters. The release of other transmitters is not excluded. Therefore, our results encourage at extending the study of MDMA biochemical profiles, in the attempt to elucidate those amphetamine-induced pathways with a potential for neurotoxicity or psycho-stimulant activity. 相似文献
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Selection of deletion mutants by polymerase chain reaction 总被引:1,自引:0,他引:1
C D Atreya P L Atreya T P Pirone 《Biochemical and biophysical research communications》1990,173(3):1344-1346
Polymerase chain reaction (PCR) based DNA amplification has replaced many time-consuming protocols in molecular biology. Here we describe a simple strategy to quickly select deletion mutants based on PCR methodology which then can be confirmed by nucleotide sequencing. A forward PCR primer is designed in such a way to recognize only the wild type sequences in the amplification reaction and thus a negative selection identifies the deletion in the samples. 相似文献
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An understanding of most plant virus disease epidemics requires a consideration of the population dynamics of insect vectors that transmit the viruses and the relationship between vector and disease dynamics. Some simple analytical (continuous-time) models are proposed for nonpersistently transmitted viruses that center on the absolute 相似文献
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F Avanzini C Alli G Bettelli F Colombo L Conforti F Pirone A Spagnoli E Taioli G Tognoni M Villella et al. 《BMJ (Clinical research ed.)》1987,294(6565):157-160
A study of blood pressure control in elderly outpatients was carried out with the participation of 444 Italian general practitioners. Of 4096 patients aged 65 years or over who were considered for recruitment, 3959 (96.7%) fulfilled all the criteria of admission and were followed up for 12 months. The findings regarding one of the aims of the study--that is, to assess the feasibility of a large scale trial in general practice--are reported. Most (87%) of the doctors completed the study. Their adherence to the protocol was highly satisfactory, leading to an acceptable quality of work. Patients'' compliance was also good; 98.6% (3898) of the patients who had fulfilled the admission criteria agreed to participate in the study, and only 4% (158) dropped out. Both of these observations support the feasibility of carrying out prospective studies in general practice. The creation of networks of general practitioners who are prepared to carry out research in their practices would allow treatment and preventive measures to be studied simply and at low cost in the appropriate setting. 相似文献
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Purification and Properties of Sugarcane Mosaic Virus Strains 总被引:1,自引:0,他引:1
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Emanuela Senatore Francesco Chiuso Laura Rinaldi Daniela Intartaglia Rossella Delle Donne Emilia Pedone Bruno Catalanotti Luciano Pirone Bianca Fiorillo Federica Moraca Giuliana Giamundo Giovanni Scala Andrea Raffeiner Omar TorresQuesada Eduard Stefan Marcel Kwiatkowski Alienke van Pijkeren Manuela Morleo Brunella Franco Corrado Garbi Ivan Conte Antonio Feliciello 《The EMBO journal》2021,40(10)
The primary cilium is a microtubule‐based sensory organelle that dynamically links signalling pathways to cell differentiation, growth, and development. Genetic defects of primary cilia are responsible for genetic disorders known as ciliopathies. Orofacial digital type I syndrome (OFDI) is an X‐linked congenital ciliopathy caused by mutations in the OFD1 gene and characterized by malformations of the face, oral cavity, digits and, in the majority of cases, polycystic kidney disease. OFD1 plays a key role in cilium biogenesis. However, the impact of signalling pathways and the role of the ubiquitin‐proteasome system (UPS) in the control of OFD1 stability remain unknown. Here, we identify a novel complex assembled at centrosomes by TBC1D31, including the E3 ubiquitin ligase praja2, protein kinase A (PKA), and OFD1. We show that TBC1D31 is essential for ciliogenesis. Mechanistically, upon G‐protein‐coupled receptor (GPCR)‐cAMP stimulation, PKA phosphorylates OFD1 at ser735, thus promoting OFD1 proteolysis through the praja2‐UPS circuitry. This pathway is essential for ciliogenesis. In addition, a non‐phosphorylatable OFD1 mutant dramatically affects cilium morphology and dynamics. Consistent with a role of the TBC1D31/praja2/OFD1 axis in ciliogenesis, alteration of this molecular network impairs ciliogenesis in vivo in Medaka fish, resulting in developmental defects. Our findings reveal a multifunctional transduction unit at the centrosome that links GPCR signalling to ubiquitylation and proteolysis of the ciliopathy protein OFD1, with important implications on cilium biology and development. Derangement of this control mechanism may underpin human genetic disorders. 相似文献