Endothelin-3 regulates neural crest cell proliferation and differentiation in the hindgut enteric nervous system

Neural crest cells (NCC) migrate, proliferate, and differentiate within the wall of the gastrointestinal tract to give rise to the neurons and glial cells of the enteric nervous system (ENS). The intestinal microenvironment is critical in this process and endothelin-3 (ET3) is known to have an essential role. Mutations of this gene cause distal intestinal aganglionosis in rodents, but its mechanism of action is poorly understood. We find that inhibition of ET3 signaling in cultured avian intestine also leads to hindgut aganglionosis. The aim of this study was to determine the role of ET3 during formation of the avian hindgut ENS. To answer this question, we created chick-quail intestinal chimeras by transplanting preganglionic quail hindguts into the coelomic cavity of chick embryos. The quail grafts develop two ganglionated plexuses of differentiated neurons and glial cells originating entirely from the host neural crest. The presence of excess ET3 in the grafts results in a significant increase in ganglion cell number, while inhibition of endothelin receptor-B (EDNRB) leads to severe hypoganglionosis. The ET3-induced hyperganglionosis is associated with an increase in enteric crest cell proliferation. Using hindgut explants cultured in collagen gel, we find that ET3 also inhibits neuronal differentiation in the ENS. Finally, ET3, which is strongly expressed in the ceca, inhibits the chemoattraction of NCC to glial-derived neurotrophic factor (GDNF). Our results demonstrate multiple roles for ET3 signaling during ENS development in the avian hindgut, where it influences NCC proliferation, differentiation, and migration.     

Discovery of imidazo[1,2-a]-, [1,2,4]triazolo[4,3-a]-, and [1,2,4]triazolo[1,5-a]pyridine-8-carboxamide negative allosteric modulators of metabotropic glutamate receptor subtype 5

Based on a hypothesis that an intramolecular hydrogen bond was present in our lead series of picolinamide mGlu₅ NAMs, we reasoned that an inactive nicotinamide series could be modified through introduction of a fused heterocyclic core to generate potent mGlu₅ NAMs. In this Letter, we describe the synthesis and evaluation of compounds that demonstrate the viability of that approach. Selected analogs were profiled in a variety of in vitro assays, and two compounds were evaluated in rat pharmacokinetic studies and a mouse model of obsessive-compulsive disorder. Ancillary pharmacology screening revealed that members of this series exhibited moderate inhibition of the dopamine transporter (DAT), and SAR was developed that expanded the selectivity for mGlu₅ versus DAT.     

连翘花的结构与繁育系统研究

通过定株观察、解剖和人工套袋交叉授粉试验对连翘花的生长发育和繁殖系统进行研究.结果表明:(1)连翘具有2种避免自交的方式,雌雄异位和雌雄异熟.其雌雄异位表现为长雄蕊短雌蕊花型和短雄蕊长雌蕊花型;雌雄异熟表现为长雄蕊短雌蕊花型为雄蕊先于雌蕊成熟,短雄蕊长雌蕊花型表现为雌蕊先于雄蕊成熟.(2)连翘花P/O值测定和户外套袋交叉授粉试验显示,P/O值为2 000±300;不同类型花的异花授粉结果率在50.64%~80.32%,其中短型花的花粉授到长型花柱头的结果率最高,达80.32%,而长型花和短型花的同型花授粉结果率分别为2.92%和34.15%,表明连翘的异花授粉结果率高于自花授粉,以异交为主,其繁育系统属兼性异交.研究结果认为,连翘雌雄异位和雌雄异熟是其自然结果率低下的主要原因,为进一步探讨连翘在木犀科中的系统进化提供了生殖生态学的依据.     

The glutamine commute: take the N line and transfer to the A

The transfer of glutamine between cells contributes to signaling as well as to metabolism. The recent identification and characterization of the system N and A family of transporters has begun to suggest mechanisms for the directional transfer of glutamine, and should provide ways to test its physiological significance in diverse processes from nitrogen to neurotransmitter release.     

A novel and highly efficient production system for recombinant adeno-associated virus vector

Recombinant adeno-associated virus (rAAV) has proven to be a promising gene delivery vector for human gene therapy. However, its application has been limited by difficulty in obtaining enough quantities of high-titer vector stocks. In this paper, a novel and highly efficient production system for rAAV is described. A recombinant herpes simplex virus type 1 (rHSV-1) designated HSV1-rc/AUL2, which expressed adeno-associated virus type2 (AAV-2) Rep and Cap proteins, was constructed previously. The data confirmed that its functions were to support rAAV replication and packaging, and the generated rAAV was infectious. Meanwhile, an rAAV proviral cell line designated BHK/SG2, which carried the green fluorescent protein (GFP) gene expression cassette, was established by transfecting BHK-21 cells with rAAV vector plasmid pSNAV-2-GFP. Infecting BHK/SG2 with HSV1-rc/AUL2 at an MOI of 0.1 resulted in the optimal yields of rAAV, reaching 250 transducing unit (TU) or 4.28×104 particles per cell. Therefore, compared     

A technique for estimating maximum harvesting effort in a stochastic fishery model

Exploitation of biological resources and the harvest of population species are commonly practiced in fisheries, forestry and wild life management. Estimation of maximum harvesting effort has a great impact on the economics of fisheries and other bio-resources. The present paper deals with the problem of a bioeconomic fishery model under environmental variability. A technique for finding the maximum harvesting effort in fluctuating environment has been developed in a two-species competitive system, which shows that under realistic environmental variability the maximum harvesting effort is less than what is estimated in the deterministic model. This method also enables us to find out the safe regions in the parametric space for which the chance of extinction of the species is minimized. A real life fishery problem has been considered to obtain the inaccessible parameters of the system in a systematic way. Such studies may help resource managers to get an idea for controlling the system.     

Detection of Glycogen-Debranching System in Trophozoites of Entamoeba histolytica

ABSTRACT. Homogenates of trophozoites of Entamoeba histolytica were shown to bring about the total degradation of glycogen while purified phosphorylase of the same source alone yielded a limit dextrin as end product. An enzyme system capable of debranching the limit dextrin was obtained from the 40,000 g pellet by extraction in aqueous medium, purified by gel filtration on Fractogel TSK HW-55(F), and separated from phosphorylase by chromatography on Blue Sepharose CL-6B and aminobutyl Agarose. The glycogen-debranching system was purified 540-fold to a state of homogeneity by criterion of disc-gel electrophoresis. The purified enzyme was able to degrade glycogen-limit dextrin in the presence of phosphorylase and exhibited activities of both amylo-1,6-glucosidase (EC 3.2.1.33) and 4- α -glucanotransferase (EC 2.4.1.25). Although amylo-1,6-glucosidase released glucose from a glycogen-phosphorylase limit dextrin, transferase activity moved single glucose residues from the limit dextrin to 4-nitrophenyl- α -glucoside yielding successively 4-nitrophenyl- α -maltoside and 4-nitrophenyl- α -maltotrioside that could be detected by HPLC. Native glycogen-debranching system exhibited a relative molecular mass of M_r= 180,000 ± 10% by gel filtration and gel electrophoresis in both denaturing and nondenaturating conditions.     

Housing of growing rabbits in individual,bicellular and collective cages: fear level and behavioural patterns

During growth (27 to 75 days of age), a total of 384 rabbits were kept in 72 individual cages, 48 bicellular cages (2 rabbits/cage) and 24 collective cages (9 rabbits/cage). To evaluate the effects of the housing system on the fear level and behavioural patterns of rabbits at the two ages (39 to 45 days and 66 to 73 days), a tonic immobility test and an open-field test were conducted and their behaviour was video recorded. In the tonic immobility test, the number of attempts to induce immobility (1.38) was lower, and the duration of immobility (47.8 s) was higher (0.05 < P < 0.01) in the rabbits housed in individual cages than in those kept in bicellular (1.72 attempts and 25.0 s of immobility) and collective cages (1.99 attempts and 25.0 s of immobility). During the open-field test, the rabbits from individual and bicellular cages showed higher latency (38.8 and 40.3 v. 27.0 s), a lower number of total (73.3 and 81.7 v. 91.9) and central displacements (3.6 and 2.8 v. 5.4) and a shorter running time (11.8 and 13.6 s v. 17.7 s) and the time biting the pen (5.5 and 9.1 s v. 28.2 s) compared with the rabbits kept in collective cages (0.05 < P < 0.001). During the 24-h video recording, the rabbits in individual and bicellular cages spent less time allogrooming (0.34% and 0.19% v. 1.44%), moving (0.74% and 0.60% v. 1.32%) and running (0.08% and 0.03% v. 0.21%) than the rabbits in the collective cages (0.01 < P < 0.001). The lowest numbers of alerts and hops were observed in the rabbits kept in bicellular cages. With increasing age, a lower number of rabbits were sensitive to the immobility test and more rabbits entered the pen spontaneously during the open-field test (P < 0.001). In conclusion, the rabbits in individual cages exhibited the highest fear level and incomplete behavioural patterns; the rabbits housed in collective cages showed the lowest fear levels and had the possibility of expressing a wider range of behaviour; and the rabbits in bicellular cages exhibited an inconsistent pattern of fear in the tonic immobility and open-field tests. Probably, these rabbits were in a less stressful condition compared with animals in individual cages because social contacts were allowed, even if freedom of movement was more limited.