Immunolocalization of the Na,K-ATPase in photoreceptor cells of the moth Manduca sexta-evidence for Na,K-ATPase on both the nonmicrovillar and the microvillar domains of the plasma membrane

Immunohistochemical and physiological studies on various insect photoreceptors have demonstrated that the Na,K-ATPase (sodium pump) is restricted to the nonreceptive nonmicrovillar area of the plasma membrane. Here, we examined the distribution of the Na,K-ATPase in photoreceptor cells of the superposition-type compound eye in the moth Manduca sexta. Using immunofluorescent and immunogold cytochemistry, we show that the Na,K-ATPase is localized to both the nonmicrovillar and the microvillar parts of the plasma membrane. Manduca photoreceptors thus deviate from the common concept that the sodium pump and the molecular components of the photoreceptive machinery reside on different domains of the plasma membrane.     

A novel potent metal-binding NDM-1 inhibitor was identified by fragment virtual,SPR and NMR screening

NDM-1 can hydrolyze nearly all available β-lactam antibiotics, including carbapenems. NDM-1 producing bacterial strains are worldwide threats. It is still very challenging to find a potent NDM-1 inhibitor for clinical use. In our study, we used a metal-binding pharmacophore (MBP) enriched virtual fragment library to screen NDM-1 hits. SPR screening helped to verify the MBP virtual hits and identified a new NDM-1 binder and weak inhibitor A1. A solution NMR study of ¹⁵N-labeled NDM-1 showed that A1 disturbed all three residues coordinating the second zinc ion (Zn2) in the active pocket of NDM-1. The perturbation only happened in the presence of zinc ion, indicating that A1 bound to Zn2. Based on the scaffold of A1, we designed and synthesized a series of NDM-1 inhibitors. Several compounds showed synergistic antibacterial activity with meropenem against NDM-1 producing K. pneumoniae.     

Reaction profile of the last step in cytochrome P-450 catalysis revealed by studies of model complexes

 A series of oxoiron(IV) porphyrin cation radical complexes was investigated as compound I analogs of cytochrome P-450. Both the spectroscopic features and the reactivities of the complexes in oxygen atom transfer to olefins were examined as a function of only one variable, the axial ligand trans to the oxoiron(IV) bond. The results disclosed two important kinetic steps – electron transfer from olefin to oxoiron(IV) and intramolecular electron transfer from metal to porphyrin radical – which are affected differently by the axial ligands. The large kinetic barrier of the latter step in the reaction of olefins with the perchlorato-bound oxoiron(IV) porphyrin cation radical complex enabled the trapping of a reaction intermediate in which the metal, but not the porphyrin radical, is reduced. The first electron transfer step is probably followed by σ-bond formation, which readily accounts for formation of isomerized organic products at low temperatures. It is finally postulated that part of the enhanced oxygenation activities of cytochrome P-450 monooxygenases and chloroperoxidases is due to a lowering of the energy barrier for the second electron transfer step via participation of their redox-active cysteinate ligand. Received: 16 January 1997 / Accepted: 24 May 1997     

Non-cytotoxic 1,2,3-triazole tethered fused heterocyclic ring derivatives display Tax protein inhibition and impair HTLV-1 infected cells

Human T cell lymphotropic virus type 1 (HTLV-1) is a human retrovirus that infects approximately 10–20 million people worldwide and causes an aggressive neoplasia (adult T-cell leukemia/lymphoma - ATL). Therapeutic approaches for the treatment of ATL have variable effectiveness and poor prognosis, thus requiring strategies to identify novel compounds with activity on infected cells. In this sense, we initially screened a small series of 25 1,2,3-triazole derivatives to discover cell proliferation inhibitors and apoptosis inducers in HTLV-1-infected T-cell line (MT-2) for further assessment of their effect on viral tax activity through inducible-tax reporter cell line (Jurkat LTR-GFP). Eight promising compounds (02, 05, 06, 13, 15, 21, 22 and 25) with activity ≥70% were initially selected, based on a suitable cell-based assay using resazurin reduction method, and evaluated towards cell cycle, apoptosis and Tax/GFP expression analyses through flow cytometry. Compound 02 induced S phase cell cycle arrest and compounds 05, 06, 22 and 25 promoted apoptosis. Remarkably, compounds 22 and 25 also reduced GFP expression in an inducible-tax reporter cell, which suggests an effect on Tax viral protein. More importantly, compounds 02, 22 and 25 were not cytotoxic in human hepatoma cell line (Huh-7). Therefore, the discovery of 3 active and non-cytotoxic compounds against HTLV-1-infected cells can potentially contribute, as an initial promising strategy, to the development process of new drugs against ATL.     

Spectroscopic investigations of intermediates in the reaction of cytochrome P450BM3–F87G with surrogate oxygen atom donors

Rapid mixing of substrate-free ferric cytochrome P450_BM3–F87G with m-chloroperoxybenzoic acid (mCPBA) resulted in the sequential formation of two high-valent intermediates. The first was spectrally similar to compound I species reported previously for P450_CAM and CYP 119 using mCPBA as an oxidant, and it featured a low intensity Soret absorption band characterized by shoulder at 370 nm. This is the first direct observation of a P450 compound I intermediate in a type II P450 enzyme. The second intermediate, which was much more stable at pH values below 7.0, was characterized by an intense Soret absorption peak at 406 nm, similar to that seen with P450_CAM [T. Spolitak, J.H. Dawson, D.P. Ballou, J. Biol. Chem. 280 (2005) 20300–20309]. Double mixing experiments in which NADPH was added to the transient 406 nm-absorbing intermediate resulted in rapid regeneration of the resting ferric state, with the flavins of the flavoprotein domain in their reduced state. EPR results were consistent with this stable intermediate species being a cytochrome c peroxidase compound ES-like species containing a protein-based radical, likely localized on a nearby Trp or Tyr residue in the active site. Iodosobenzene, peracetic acid, and sodium m-periodate also generated the intermediate at 406 nm, but not the 370 nm intermediate, indicating a probable kinetic barrier to accumulating compound I in reactions with these oxidants. The P450 ES intermediate has not been previously reported using iodosobenzene or m-periodate as the oxygen donor.     

Flame retardant finishing of cotton fabric based on synergistic compounds containing boron and nitrogen

Boric acid and compound containing nitrogen, 2,4,6-tri[(2-hydroxy-3-trimethyl-ammonium)propyl]-1,3,5-triazine chloride (Tri-HTAC) were used to finish cotton fabric. The flame retardant properties of the finished cotton fabrics and the synergetic effects of boron and nitrogen elements were investigated and evaluated by limited oxygen index (LOI) method. The mechanism of cross-linking reaction among cotton fiber, Tri-HTAC, and boric acid was discussed by FTIR and element analysis. The thermal stability and surface morphology of the finished cotton fabrics were investigated by thermogravimetric analysis (TGA) and scanning electron microscope (SEM), respectively. The finishing system of the mixture containing boron and nitrogen showed excellent synergistic flame retardancy for cotton fabric. The cotton fabric finished with mixture system had excellent flame retardancy. The LOI value of the treated cotton fabric increased over 27.5. Tri-HTAC could form covalent bonds with cellulose fiber and boric acid. The flame retardant cotton fabric showed a slight decrease in tensile strength and whiteness. The surface morphology of flame retardant cotton fiber was smooth.     

Insights on the Behavior of Late Paleozoic Aquatic Crustaceans (Pygocephalomorpha?): Compound Trace Fossils from Western Argentina

We describe intergradations between the arthropod repichnial trace fossils Diplichnites gouldi (Gevers et al., 1971), Dendroidichnites Demathieu et al., 1992, and Monomorphichnus Crimes, 1970 with two distinct cubichnial traces, Gluckstadtella cooperi Savage, 1971 and Huilmuichnus santracruzensis new ichnogenus and ichnospecies. These compound ichnofossils are part of high ichnodiversity arthropod-dominated trace fossil associations from the Late Carboniferous Agua Escondida Formation of the San Rafael Basin (Argentina). The complex ichnotaxonomy of Diplichnites and the ichnospecies D. gouldi are discussed and some possible working solutions are proposed. The most likely producer of the compound trace fossils are the Pygocephalomorpha, a group of Late Paleozoic crustaceans that are recorded in nearby basins from Uruguay and Brazil. The compound trace fossils are used to infer a number of complex behaviors of pygocephalomorphs in a subaqueous setting, including caridoid (tail flip) or more primitive escape reactions, landing followed by resting and transient resting during normal walking. In addition, different gaits can be inferred from well-preserved D. gouldi, which can be correlated with contrasting trackway morphology.     

福建甘薯病毒病病原鉴定及主要病毒多样性

【背景】病毒病是甘薯的一种重要病害,给甘薯生产带来了严重的经济损失,而生产中甘薯病毒病病原种类复杂多样。【目的】明确福建甘薯病毒病种类、分布及流行,对主要病毒进行多样性分析。【方法】从福建主要甘薯种植区采集病毒病样品,利用PCR/RT-PCR的方法对采集的样品进行病原检测,获得病毒序列,利用MEGA 6.0构建系统进化树进行遗传分析。【结果】从福建7个甘薯产区鉴定12种甘薯病毒,包括9种RNA病毒:甘薯羽状斑驳病毒(Sweet potato feathery mottle virus,SPFMV)、甘薯褪绿矮化病毒(Sweet potato chlorotic stunt virus,SPCSV)、甘薯G病毒(Sweet potato virus G,SPVG)、甘薯C病毒(Sweet potato virus C,SPVC)、甘薯2号病毒(Sweet potato virus 2)、甘薯褪绿斑病毒(Sweet potato chlorotic fleck virus,SPCFV)、甘薯潜隐病毒(Sweet potato latent virus,SPLV)、甘薯轻型斑点病毒(Sweetpotatomildspeakingvirus,SPMSV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV),3种DNA病毒:甘薯卷叶病毒(Sweet potato leaf curl virus,SPLCV),甘薯无症状1号病毒(Sweet potato symptomless virus 1,SPSMV-1),甘薯杆状DNA病毒B (SPBV-B)。SPFMV、SPCSV、SPVG和SPLCV检出率最高,分别为50.28%、41.90%、35.75%和24.58%,CMV检出率最低,为2.79%,未检测到甘薯C-6病毒(SweetpotatoC-6)和甘薯轻型斑驳病毒(Sweetpotatomild mottle virus,SPMMV)。福建甘薯主要以2-6种病毒复合侵染为主,单一侵染率占14.39%,2种以上复合侵染占85.61%。福建SPFMV分离物存在EA、O和RC3种株系,SPCSV分离物存在WA1种株系,未发现EA株系,甘薯卷叶病毒分属于2个不同的株系群。【结论】福建甘薯病毒种类多样,以复合侵染为主,且存在多种株系,遗传结构复杂。     

Intracellular recording from a spider vibration receptor

The present study introduces a new preparation of a spider vibration receptor that allows intracellular recording of responses to natural mechanical or electrical stimulation of the associated mechanoreceptor cells. The spider vibration receptor is a lyriform slit sense organ made up of 21 cuticular slits located on the distal end of the metatarsus of each walking leg. The organ is stimulated when the tarsus receives substrate vibrations, which it transmits to the organ’s cuticular structures, reducing the displacement to about one tenth due to geometrical reasons. Current clamp recording was used to record action potentials generated by electrical or mechanical stimuli. Square pulse stimulation identified two groups of sensory cells, the first being single-spike cells which generated only one or two action potentials and the second being multi-spike cells which produced bursts of action potentials. When the more natural mechanical sinusoidal stimulation was applied, differences in adaptation rate between the two cell types remained. In agreement with prior extracellular recordings, both cell types showed a decrease in the threshold tarsus deflection with increasing stimulus frequency. Off-responses to mechanical stimuli have also been seen in the metatarsal organ for the first time.     

复方丹参注射液联合阿奇霉素治疗小儿喘息性支气管炎的效果及对炎性因子的影响

目的:探讨复方丹参注射液联合阿奇霉素治疗小儿喘息性支气管炎的效果及对炎性因子的影响。方法:选取2015年6月～2018年6月我院收治的喘息性支气管炎患儿300例,采用随机数字表法将患儿分为两组,每组各150例。对照组在常规治疗的基础上给予阿奇霉素注射液治疗,观察组在对照组的基础上联合应用复方丹参注射液治疗。比较两组的临床治疗效果,临床症状缓解时间及住院时间,治疗前后两组血清白介素(Interleukin, IL)-6、IL-8和肿瘤坏死因子-α(Tumor necrosis factor-α, TNF-α)水平的变化情况及不良反应发生情况和复发率。结果:治疗后,观察组治疗总有效率显著高于对照组(93.33%VS.85.33%, P0.05);观察组喘息缓解时间、咳嗽缓解时间、哮鸣音消失时间、体温恢复时间及住院时间均显著短于对照组(P0.05);两组治疗后血清IL-6、IL-8和TNF-α水平均较治疗前显著下降,且观察组更低(P0.05);两组不良反应发生率比较差异无统计学意义(P0.05),观察组的复发率显著低于对照组(P0.05)。结论:复方丹参注射液联合阿奇霉素可快速缓解喘息性支气管炎患儿的临床症状、体征并缩短住院时间,提高临床治疗效果,且复发率低,安全性较高,这可能与其可显著降低患儿血清IL-6、IL-8和TNF-α水平有关。