On the dynamics of electrically-coupled neurons with inhibitory synapses

We study the dynamics and bifurcations of noise-free neurons coupled by gap junctions and inhibitory synapses, using both delayed delta functions and alpha functions to model the latter. We focus on the case of two cells, as in the studies of Chow and Kopell (2000) and Lewis and Rinzel (2003), but also show that stable asynchronous splay states exist for globally coupled networks of N cells dominated by subthreshold electrical coupling. Our results agree with those of Lewis and Rinzel (2003) in the weak coupling range, but our Poincaré map analysis yields more information about global behavior and domains of attraction, and we show that the explicit discontinuous maps derived using delayed delta functions compare well with the continuous history-dependent, implicitly-defined maps derived from alpha functions. We find that increased bias currents, super-threshold electrical coupling and synaptic delays promote synchrony, while sub-threshold electrical coupling and fast synapses promote asynchrony. We compare our analytical results with simulations of an ionic current model of spiking cells, and briefly discuss implications for stimulus response modes of locus coeruleus and for central pattern generators. Action Editor: F. Skinner     

Chemical composition and antifungal efficacy of five essential oils against Calonectria pseudonaviculata,the causal agent of boxwood blight: an in vitro study

Recent epidemics of box (boxwood) blight caused by the fungus Calonectria pseudonaviculata in Hyrcanian forests led to the search for eco-friendly fungicides. Essential oils (EOs) of medicinal plants undoubtedly have been highlighted as a new therapeutic approach. In this study, EOs from Cuminum cyminum, Lavandula angustifolia, Thymus daenensis, Trachyspermum copticum and Hyssopus officinalis were evaluated for their inhibitory activities against C. pseudonaviculata Cy-08 in vitro. Quantitative and qualitative composition variations of the EOs were also analysed by GC–MS. Based on the high inhibitory efficacy and low related EC₅₀, MFC and MIC values, T. copticum, L. angustifolia and Th. daenensis EOs can be mentioned as those providing extraordinary efficacy against Cy-08, respectively. Major chemical components of T. copticum, L. angustifolia and Th. daenensis EOs were thymol, linalyl acetate and thymol, respectively. Thymol and linalyl acetate may be the most effective compounds against C. pseudonaviculata.     

兴奋性突触传入对皮层神经元形态发育的影响

目的:探讨离子型谷氨酸受体中的AMPA受体(α-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor, AMPA受体)和NMDA受体(N-methyl-D-aspartic acid receptor)对抑制性中间神经元以及兴奋性神经元的形态发育的影响。方法:采用原代培养皮层神经元,通过药物干预AMPA受体和/或NMDA受体的方法阻断神经元的离子型谷氨酸受体,并采用GAD67-GFP鼠的绿色荧光来显示混合细胞群中抑制性神经元、CaMKII免疫荧光染色显示兴奋性神经元。结果:当阻断AMPA和/或NMDA受体时,光镜下显示神经元网络的密度降低,且随着药物浓度的增加,神经元网络的变化更明显。对于GFP阳性的抑制性神经元,当阻断AMPA受体时,神经元突起分支数降低至对照组的约65%(低浓度)和55%(高浓度),突起长度缩短至对照组的大约43%(低浓度)和36%(高浓度);当阻断NMDA受体时,分支数降低至约70%(低浓度)和45%(高浓度),长度缩短至约43%(低浓度)和31%(高浓度);联合用药时,分支数和长度分别为对照的约42%和38%。对于CaMKII阳性的兴奋性神经元,尽管变化程度稍弱,但其形态也出现类似变化。当阻断AMPA受体时,神经元的分支数降低至对照组的64%(高浓度),突起长度变化不大;当阻断NMDA受体时,分支数降低至约50%(高浓度),长度缩短至约77%(低浓度)和71%(高浓度);联合用药时,分支数和长度分别为对照的约69%和62%。结论:在神经元发育的过程中,离子型谷氨酸受体介导的兴奋性突触传入可影响抑制性神经元和兴奋性神经元的形态发育,最终对神经环路的形成发挥重要的调控作用。     

In vitro effect of important herbal active constituents on human cytochrome P450 1A2 (CYP1A2) activity

This study was designed to investigate eight herbal active constituents (andrographolide, asiaticoside, asiatic acid, madecassic acid, eupatorin, sinensetin, caffeic acid, and rosmarinic acid) on their potential inhibitory effects on human cytochrome P450 1A2 (CYP1A2) activity. A fluorescence-based enzyme assay was performed by co-incubating human cDNA-expressed CYP1A2 with its selective probe substrate, 3-cyano-7-ethoxycoumarin (CEC), in the absence or presence of various concentrations of herbal active constituents. The metabolite (cyano-hydroxycoumarin) formed was subsequently measured in order to obtain IC₅₀ values. The results indicated that only eupatorin and sinensetin moderately inhibited CYP1A2 with IC₅₀ values of 50.8 and 40.2 μM, while the other active compounds did not significantly affect CYP1A2 activity with IC₅₀ values more than 100 μM. K_i values further determined for eupatorin and sinensetin were 46.4 and 35.2 μM, respectively. Our data indicated that most of the investigated herbal constituents have negligible CYP1A2 inhibitory effect. In vivo studies however may be warranted to ascertain the inhibitory effect of eupatorin and sinensetin on CYP1A2 activity in clinical situations     

椪柑精油及其主要抑菌组分对菌核青霉的抑制作用

采用水蒸气蒸馏法提取椪柑(Citrus reticulate Blanco)果皮精油,并用GC-MS对其成分进行分析,共鉴定出46种成分,主要包括柠檬烯(57.67%)、β-芳樟醇(5.36%)、2-蒈烯(4.47%)、β-蒎稀(4.31%)、γ-松油烯(4.08%)、α-蒎烯(3.27%)、1,2-二异丙烯基环丁烷(2.87%)、β-月桂烯(2.77%)、α-侧柏烯(2.40%)、β-水芹烯(2.24%)、癸醛(1.80%)和香茅醇(1.49%)等。采用污染食物技术(poisoned food technique)和液体培养法(liquid culture)测定了不同浓度椪柑精油以及主要抑菌组分对菌核青霉的抑制作用。结果表明:不同浓度(0.5~20μl/ml)的椪柑精油对菌核青霉生长均有一定的抑制,且抑菌效果与浓度呈正相关。无论是采用污染食物技术还是液体培养法,20μl/ml椪柑精油均能完全抑制菌核青霉生长;随着培养 时间延长到7d, 20μl/ml椪柑精油抑菌效果有所下降,但仍能显著抑制菌核青霉生长。采用污染食物技术考察了椪柑精油中7种常见抑菌成分对菌核青霉的影响。结果表明,0.04μl/ml柠檬醛和1.07μl/ml β-芳樟醇能显著抑制菌核青霉生长,而其他组分无明显作用。实验结果表明,椪柑精油的抑菌作用可能归功于其所含的柠檬醛和β-芳樟醇。     

解淀粉芽孢杆菌KN-BL-1及其发酵豆粕产抗菌肽类物质的研究

以牛津杯法比较解淀粉芽孢杆菌KN-BL-1及其发酵豆粕的抑菌效果,通过酸沉淀法分离粗多肽,确定产生抑菌效果的为多肽类物质,通过DEAE-Sepharose离子交换层析及Sephadex G-25凝胶过滤层析分离纯化粗多肽以研究其各组分的抑菌效果,并使用4000 Q TRAP液相色谱-质谱联用仪初步确定抗菌肽样品的分子量范围.结果表明,KN-BL-1及其发酵豆粕浸提液对S.aureus等革兰氏阳性菌均有明显的抑菌效果;粗多肽经过层析分离纯化后,三个峰表现抑菌特性,其中有一个抑菌效果最好,抑菌圈清晰,直径为19.8 mm.     

Characteristics and inhibition by flavonoids of 20alpha-hydroxysteroid dehydrogenase activity in mouse tissues

Progesterone was stereoselectively reduced to a metabolite 20alpha-hydroxy-4-pregnen-3-one in the cytosolic fraction from the liver of male mice, indicating that the reduction of progesterone is catalyzed by 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD). The cytosolic 20alpha-HSD activity was observed not only in the liver, but also in the kidney and lung. In liver cytosol, both NADPH and NADH were effective as cofactors for 20alpha-HSD activity, although NADPH was better than NADH for the enzyme activity. On the other hand, 20alpha-HSD activity in kidney cytosol required only NADPH as a cofactor. No significant sex-related difference of 20alpha-HSD activity was observed in liver and kidney cytosols. Flavonoids have been reported to inhibit the biosynthesis and metabolism of steroids. However, little is known about inhibitory effects of flavonoids on 20alpha-HSD activity. Thus, the effects of 16 flavonoids on 20alpha-HSD activity were examined, using liver cytosol of male mice. Among flavonoids tested, fisetin, apigenin, naringenin, luteolin, quercetin and kaempferol exhibited high inhibitory potencies for the 20alpha-HSD activity. We propose the possibility that these flavonoids augment progesterone signaling by inhibiting potently 20alpha-HSD activity in non-reproductive tissues.     

A novel family of diversified immunoregulatory receptors in teleosts is homologous to both mammalian Fc receptors and molecules encoded within the leukocyte receptor complex

Three novel and closely related leukocyte immune-type receptors (IpLITR) have been identified in channel catfish (Ictalurus punctatus). These receptors belong to a large polymorphic and polygenic subset of the Ig superfamily with members located on at least three independently segregating loci. Like mammalian and avian innate immune regulatory receptors, IpLITRs have both putative inhibitory and stimulatory forms, with multiple types coexpressed in various lymphoid tissues and clonal leukocyte cell lines. IpLITRs have an unusual and novel relationship to mammalian and avian innate immune receptors: the membrane distal Ig domains of an individual IpLITR are related to fragment crystallizable receptors (FcRs) and FcR-like proteins, whereas the membrane proximal Ig domains are related to several leukocyte receptor complex encoded receptors. This unique composition of Ig domains within individual receptors supports the hypothesis that functionally and genomically distinct immune receptor families found in tetrapods may have evolved from such ancestral genes by duplication and recombination events. Furthermore, the discovery of a large heterogeneous family of immunoregulatory receptors in teleosts, reminiscent of amphibian, avian, and mammalian Ig-like receptors, suggests that complex innate immune receptor networks have been conserved during vertebrate evolution.Electronic supplementary material Supplementary material is available for this article at and is accessible for authorized users. GenBank Submissions: The sequences presented in this article have been submitted to GenBank under the following accession numbers: AAW82352, IpLITR1; AAW82353, IpLITR2; AAW82354, IpLITR3.     

Molecular cloning and expression of a novel Kazal-type serine proteinase inhibitor gene from Zhikong scallop Chlamys farreri, and the inhibitory activity of its recombinant domain

Serine proteinase inhibitors (SPIs) play important roles in host physiological and immunological processes in all multicellular organisms. A novel Kazal-type SPI gene was cloned from the Zhikong scallop Chlamys farreri (designated as CfKZSPI) by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of CfKZSPI was of 1788 nucleotides with a canonical polyadenylation signal sequence AATAAA and a polyA tail, and an open reading frame (ORF) encoding a polypeptide of 509 amino acids with a putative signal peptide of 22 amino acids. The deduced amino acid sequence of CfKZSPI contained 12 tandem Kazal domains with high similarity to other Kazal-type SPIs. The temporal expression of CfKZSPI in hemocytes after Vibrio anguillarum challenge was recorded by quantitative real-time RT-PCR. The relative mRNA expression level of CfKZSPI was up-regulated and reached 43.6-fold at 3h post-challenge. After a decrease at 6h, the expression level increased again and reached 207.8-fold at 12h post-challenge. The 12th Kazal domain of CfKZSPI was recombined into pET-32a(+) and expressed in Escherichia coli Rosetta-gami (DE3) to investigate its inhibitory activity. The purified recombinant protein (rCfKZSPI-12) showed significant inhibitory activity against trypsin but no activity against thrombin. When the molar ratio of inhibitor to trypsin reached 1:1, almost 90% of the enzyme activity could be inhibited, which suggested that one molecule of rCfKZSPI-12 was able to inhibit one molecule of trypsin. Kinetics analysis with Dixon plot showed that the inhibition constant (Ki) of rCfKZSPI-12 to trypsin was 173 nmol L(-1). These results indicated that CfKZSPI was a novel Kazal-type SPI with significant inhibitory activity against trypsin, and was suspected to be involved in scallop immune response.     

The characteristics of nutrient removal and inhibitory effect of Ulva clathrata on Vibrio anguillarum 65

To investigate the ecological effect of macroalgae on de-eutrophication and depuration of mariculture seawater, the variation of dissolved inorganic nitrogen (DIN) and phosphate (DIP), the amount of Vibrio anguillarum, and total heterotrophic bacteria in Ulva clathrata culture, as well as on the algal surface, were investigated by artificially adding nutrients and V. anguillarum strain 65 from February to April 2006. The results indicated that U. clathrata not only had strong DIN and DIP removal capacities, but also showed a significant inhibitory effect on V. anguillarum, although not reducing the total heterotrophic bacteria. Vibrio anguillarum 65 dropped from 5∼8 × 10⁷ cfu mL⁻¹ to 10 cfu mL⁻¹ (clone-forming units per mL) in 10 g L⁻¹ of fresh U. clathrata culture within 2 days; i.e., almost all of the Vibrios were efficiently eradicated from the algal culture system. Our results also showed that the inhibitory effect of U. clathrata on V. anguillarum strain 65 was both DIN- and DIP-dependent. Addition of DIN and DIP could enhance the inhibitory effects of the algae on the Vibrio, but did not reduce the total heterotrophic bacteria. Further studies showed that the culture suspension in which U. clathrata was pre-cultured for 24 h also had an inhibitory effect on V. anguillarum strain 65. Some unknown chemical substances, either released from U. clathrata or produced by the alga associated microorganisms, inhibited the proliferation of V. anguillarum 65.