Glycine origin of the methyl substituent on C7′-N of octodiose for the biosynthesis of apramycin

Apramycin is unique in the aminoglycoside family due to its octodiose moiety. However, either the biosynthesis process or the precursors involved are largely unknown. Addition of glycine, as well as serine or threonine, to the Streptomyces tenebrabrius UD2 fermentation medium substantially increases the production of apramycin with little effect on the growth of mycelia, indicat-ing that glycine and/or serine might be involved in the biosynthesis of apramycin. The ¹³C-NMR analysis of [2-¹³C] glycine-fed (25% enrichment) apramycin showed that glycine specifically and efficiently incorporated into the only N-CH₃ substituent of apramycin on the C7′ of the octodiose moiety. We noticed that the in vivo concentration of S-adenosyl methionine increased in parallel with the addition of glycine, while the addition of methione in the fermentation medium significantly decreased the productivity of apramycin. Therefore, the methyl donor function of glycine is proposed to be involved in the methionine cycle but methionine itself was proposed to inhibit the methylation and methyl transfer processes as previously reported for the case of rapamycin. The ¹⁵N NMR spectra of [2-¹³C,¹⁵N]serine labeled apramycin indicated that serine may also act as a limiting precursor contributing to the ―NH₂ substituents of apramycin.     

Determination of Mercury and Methylmercury in Hair of the Czech Children’s Population

The developed method for mercury speciation analysis has been validated and used for the biomonitoring study of mercury species in human hair. Statistical evaluation proved the reliability of simplified determination of inorganic mercury (difference between total mercury and methylmercury). The results of the validation showed that the method is very well suitable for the determination of both species of mercury in hair for biomonitoring purposes. Non-exposed schoolchildren from three areas in the western and central part of the Czech Republic were chosen as the target group. Tenth of a microgram per gram of the total mercury were generally found in the analyzed hair; values higher than 1 μg g⁻¹ were detected only exceptionally. Comparable results were obtained for two western areas and differed significantly from those for the third area located in the central part of the Czech Republic. In the areas examined, the mean methylmercury contents amounted to 23–46% of the total mercury in the hair. The results confirm an assumption that exposure to mercury does not pose a significant risk to the population in the Czech Republic.     

A Scientist Guerilla Fighter in the Frontiers of Bioinformatics——In Memory of Bailin Hao

正The well-known physicist and bioinformaticist, Professor,Academician of Chinese Academy of Sciences (the Academy),and the former director of Institute of Theoretical Physics (the Institute; 1990–1994), Mr. Bailin Hao, passed away early this year (March 7, 2018). In the cover of this issue of Genomics,ProteomicsBioinformatics (GPB), the Editorial Board has decided to publish his art-processed portrait in memory of     

Importance of functional classification in the use of carabids for the environmental risk assessment of the GE crops and other agricultural practices

Carabids(Coleoptera:Carabidae)seem to be suitable bioindicalors of the environmental impacts of novel agrotechnologies,including deployment of the genetically engineered(GE)crops.In this article,we describe our effort to employ carabids in the environmental risk assessment(ERA).GE maize MON88017,its near-isogenic hybrid nontreated or treated with the soil insccticide chlorpyrifos,and two reference hybrids were used to compare three different ways how to utilize carabids in ERA.The analysis of abundance of all captured carabids or of the most abundant carabid specics did not disclose any differences between the treatments.The analysis based on the categories of functional traits revealed distinct fecatures of some treatments and proved suitable for ERA because it permitted field data transportability in spite of different species compositions.Our results indicate that GE maize has no detrimental environmental effecl,On the other hand,we found significant trends toward lower abundance and lower species number(including analysis of all carabid species together)in plots treated with the insecticide,and some tendencies to higher abundance and higher species number in plots sown with the reference hybrid PR38N86.Using functional group indicators alows identification of unintended changes in ecological functions of agroecosystem and comparability across geographics.We recommend data evaluation at the level of the categories of functional traits in ERA of GE crops and other agricultural practices.     

Two complexes at the site of microtubule nucleation

<正>Cortical microtubule(MT)arrays are dynamic filamentous structures that are essential for cell differentiation and development in plants.However,the molecular mechanisms that control the organization of cortical MT arrays are not well understood.Early studies have revealed that the formation of cortical MT arrays involves MT nucleation on existing cortical MTs.The growth of new MTs follows the polarity of existing MTs and the orientation of new MTs is either in parallel with extant MTs or at a small angle(about40 degree)to the extant MTs[1].Nucleation machinery appears to be conserved between animals and plants in     

A legacy of the “1% program”-The “Chinese Chapter” of the human genome reference sequence

正The year of 2018 marks the 65th anniversary of the discovery of DNA double helix and the 15th anniversary of the successful completion of the international Human Genome Project (HGP),the two revolutions in life sciences (Sharp, 2014). The year also sees the effective implementation of the "1%Program", the Chinese contribution to the reference sequence of the human genome,which coincides with the 40th anniversary of the founding of the Genetics Society of China, one of the most influential national academic organizations in China.     

The effect of the type of closure on the gas composition of the headspace and the growth of GF 677 peach × almond rootstock cell suspension cultures

Summary The influence of culture flask closures, i.e., cotton plugs and rubber and aluminum-foil caps, on headspace gas composition and growth of leaf petiole callus-derived GF 677 cell suspensions was comparatively tested. Oxygen concentration always remained comparable to that of the lab atmosphere and CO₂ and C₂H₄ levels were slightly higher when flasks were closed with cotton plugs. By contrast, the gas environment markedly changed within 72 to 96 h in culture inside rubber and aluminum-capped flasks. Under rubber caps, CO₂ increased up to 18%, with a net production of about 0.8 mmol CO₂, oxygen decreased to 6% within 72 h, and ethylene accumulated up to 9 μl liter⁻¹ after 96 h. When aluminum foil closures were used, C₂H₄ and CO₂ increased over the first 72 h, reaching concentrations of about 6 μl liter⁻¹ and 7% (0.3 mmol produced), respectively, and decreasing after 192 h to 0.1 μl⁻¹ and 2%, respectively. The gaseous environment markedly affected cell growth. The exponential growth period of suspensions cultured in flasks closed with cotton plugs and aluminum foil caps started after about 72 h and the stationary phase after 240 h, the cell dry weight reaching its maximum at about threefold the initial weight. Large cell aggregates were found in flasks closed with cotton plugs, slight aggregation with aluminum closures, and no aggregates with rubber caps. However, hardly any growth, progressive browning of the suspensions, and the death of most cells occurred in rubber-capped flasks. A type of closure allowing low gas exchange rates, like aluminum caps, and frequent subcultures thus seems most conducive to rapid growing, more homogeneous GF 677 cell suspension cultures, and the prevention of aggregates, if undesired.     

Melatonin in the seasonal response of the aphid Acyrthosiphon pisum

Aphids display life cycles largely determined by the photoperiod.During the warm long-day seasons.most aphid species reproduce by viviparous parthenogenesis.The shortening of the photoperiod in autumn induces a switch to sexual reproduction.Males and sexual females mate to produce overwintering resistant eggs.In addition to this full life cycle(holocycle),there are anholocyelic lineages that do not respond to changes in photoperiod and reproduce continuously by parthenogenesis.The molecular or hormonal events that trigger the scasonal response(i.c,induction of the sexual phenotypes)are still unknown.Although circadian synthesis of melatonin is known to play a key role in vertebrate photoperiodism,the involvement of the circadian clock and/or of the hor-mone melatonin in insect seasonal responses is not so well established.Here we show that melatonin levels in the aphid Acyrthosiphon pisum are significantly higher in holocyclice aphids reared under short days than under long days,while no differences were found between anholoeyelic aphids under the same conditions.We also found that melatonin is localized in the aphid suboesophageal ganglion(SOG)and in the thoracic ganglionic mass(TGM).In analogy to vertcbrates,insect-type arylalkxylamine N-acetyltransferases(i-AANATs)are thought to play a key role in melatonin synthesis.We measured the expression of four I-AANAT genes identified in A.pisum and localized two of them in situ in the insect central nervous systems(CNS).Levels of expression of these genes were compatible with the quantities of melatonin observed.Moreover,like melatonin,expression of these genes was found in the SOG and the TGM.     

Expression and intercellular trafficking of the VP22 protein of CVI988/Rispens vaccine strain of Marek’s disease virus

The viral protein 22 (VP22) in the tegument of Marek’s disease virus serotype 1 (MDV-1) plays an im-portant role in cell-to-cell spread and viral propagation. Antiserum against the carboxyl terminus of VP22 was prepared by immunizing mice with recombinant VP22 expressed in E. coli, and used to in-vestigate its expression in chicken embryo fibroblast (CEF) cells infected with different MDV-1 strains. At an infection dose of PFU=50, intercellular trafficking of the VP22 into the nuclei of the surrounding receipt cells was detected as early as 3 hours post infection. By 6 hours after infection (before viral plague formation), the protein was detected in the whole nuclei of the recipient cells with no difference among MDV-1 strains CVI988/Rispens, GA and RB1B. Intra-nuclear accumulation of the VP22 protein was further increased when the viral plagues started to form. These results indicate that, albeit the ex-istence of the ²⁰¹TKSERT²⁰⁶ deletion, the VP22 of the CVI988/Rispens vaccine strain has also intercel-lular-trafficking function, which might serve as a potential alternative delivering protein instead of virulent strains VP22.     

Construction of normal human IgE phage antibody library and the screening of the anti—trichosanthin IgE

Allergen specific IgE response is the major cause of immediate hypersensitivity.However the number of IgEproducing B cells and the amount of IgE,especially the specific IgE,are so low,it greatly impedes the study of the allergic-specifc antibody responses.Here we report the construction of a normal human IgE combinatorial library.The repertoire of IgE VH genes and of κ genes were separately amplified from normal human peripheral blood lymphocytes through RT-PCR,and were then constructed to form the phage surface display human Fab(IgEVH) library.A plant protein allergen,trichosanthin(TCS),was used to affinity-enrich and to screen the anti-TCS phage HuFab clones from the library.Human IgE(Fab) to TCS were detected.     

Use of Whole-Exome Sequencing for the Diagnosis of Atypical Birte-Hogge-Dubé Syndrome

正Destruction of the lung parenchyma may result in the formation of cysts,cavities and bullae.Definition of the type of destructive lesion is best achieved by high-resolution computed tomography(HRCT)scans of the chest(TrotmanDickenson,2014).The loss of lung parenchyma may result     

Effect of ConA—receptor interaction on the structure of cell membrane

Recently,the effect of ligand receptor interaction on the membrane structure of liposomes has been studied extensively,However,little is known about how it exists on biological membranes,In this paper,the effect of Concanavalin A(ConA) receptorinteratcion on the structure of cell membranes was studied by Circular DIchrosim(CD) and 31P Nuclear Magnetic Resonance(NMR).CD results of both the purified macrophage membranes and human erythrocyte hgosts(EG) showed that the conformation of membrane proteins changed after ConA binding.For further research,31P-NMR was used to detect the orgainzation of phosp[holipid molecules on macrophage membranes.After ConA binding,the tendercy to form non bilayer structure increased with the amount of ConA.The changes of 31P-NMR spectra of living macrophages might be partly due to the above stated reason too.In addition,ConA-receptor interaction also induced similar results of 31P-NMR spectra in EG.In contrast,wheat germ agglutinin (WGA),another kind of lectin,rarely showed the same influence.     

Val 70,Phe 72 and the last seven amino acid residues of C—terminal are essential to the function of norepinephrine transporter

The norepinephrine transporter(NET) is a member of the Na^ /Cl^- dependent neurotransmitter transporter family and constitutes the target of several clinically important antidepressants.To delineate the critical amino acid residues and the function of C-terminal in regulating transport activity of NET,here we constructed two site mutants (V70F,F72V;V70I,F72V) and one C-terminal truncated mutant (Δ 611-617).The wild type and mutants of NET were expressed in Xenopus oocytes by injection of their cRNA.We found that all of these mutants lost their transport activity.These results indicate that the amino acid residues of V70 and F72,and the last seven amino acids of C-terminal are essential to the transport activity of NET.     

Histochemical study of the pre—and postnatal development of acetylcholinesterase in the rat spinal cord

The distribution of acetylcholinesterase(AChE)-positive structures in the developing rat spinal cord was studied with AChE-histochemistry.AChE-positive perikarya were first seen on embryonic day 14(E14) in the ventrolateral portion of the spinal cord.From that time onward.AChE=containing cells appeared gradually in the intermediate gray,dorsal horn and lateral spinal nucleus of the spinal cord in a ventral-to-dorsal,and lateral-to-medial order.No obvious rostral-to-caudal sequence was found.At birth,the distribution pattern of AChE-positive perikarya was basically similar to that in adults.After birth a dramatic increase in the AChE staining intensity extended from postnatal day 5(P5) to postnatal day 21(P21),In addition,two phases of transient AChE staining were observed in the external surface of the dorsal horn from embryonic day 15(E15) to embryonic day 21(E21) and in the marginal layer from embryonic day 21(E21) to postnatal day 14(P14),respectively.     

The myth of “minima” and “maxima”, the species of Physalis in the Indian Subcontinent

The status of the names, Physalis minima L. and P. maxima Mill. (Solanaceae), and their alleged presence on the Indian subcontinent are discussed. The issues of nativity and identity of Linnaean Physalis minima are long-debated while the use of the name P. maxima Mill. and its report from India are recent. The available evidence indicates that the name "P. minima L." is misapplied to two different elements, viz., P. angulata L. and P. lagascae Roem. & Schult. The name Physalis minima L. may be rejected as nomen confusum, for which the paper provides the primary information. As on today, it is submerged under the synonymy of P. angulata L. The correct name for the widely known P. minima is P. lagascae. The name "P. maxima Mill." applied to the escape and naturalized weed in the Indian subcontinent and elsewhere is to be substituted by P. pruinosa L., a name misapplied to P. grisea (Waterf.) M. Martínez.     

Survival strategies of some species of the phytoplankton community in the Barra Bonita Reservoir (São Paulo,Brazil)

The dynamics of the phytoplankton community in the Barra Bonita Reservoir (S?o Paulo, Brazil) were studied through daily sampling in the field (integrated samples from the euphotic zone) and microcosm experiments, for two short periods: the winter of 1993 (June 30 to July 10) and the summer of 1994 (January 24 to February 2). The goal of the study was to evaluate and compare the variations in the composition of isolated phytoplankton community which occur over short periods of time. Three series were separated into Erlenmeyer flasks for each study period, with samples from the euphotic zone divided into three portions: total, smaller than 64 μm, and smaller than 20 μm. All of the Erlenmeyer flasks were inclubated in situ at the sampling station. The maximum period of incubation was 10 d. Variations of the community in the euphotic zone were characterised by high diversity and a community in a state of non-equilibrium in winter, without the predominance of any species. In the summer, the community presented a low diversity and a state of equilibrium, with the predominance of Microcystis aeruginosa. The microcosm experiments showed that the confinement of the community in the Erlenmeyer flasks, and therefore in isolation from the physical variability of the ecosystem, especially in relation to the mixing patterns, stimulated the return of the community to the initial phases of succession with the predominance of small species and those which grow rapidly (r-selective or C-strategist). This revised version was published online in July 2006 with corrections to the Cover Date.     

Different Effects of Homocysteine and Oxidized Low Density Lipoprotein on Methylation Status in the Promoter Region of the Estrogen Receptor α Gene

We investigated the effects of homocysteine (Hcy) and oxidized low density lipoprotein (ox-LDL) on DNA methylation in the promoter region of the estrogen receptor α (ERos) gene,and its potentialmechanism in the pathogenesis of atherosclerosis.Cultured smooth muscle cells (SMCs) of humans weretreated by Hcy and ox-LDL with different concentrations for different periods of time.The DNA methylationstatus was assayed by nested methylation-specific polymerase chain reaction,the lipids that accumulated inthe SMCs and foam cell formations were examined with Oil red O staining.The proliferation of SMCs wasassayed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.The results showedthat ox-LDL in moderate concentrations (10-40 mg/L) induced de novo methylation in the promoter regionof the ERα gene of SMCs.However,high concentrations (50 mg/L) of ox-LDL,resulted in demethylation ofERα.The Hcy treatment resulted in de novo methylation in the promoter region of the ERα gene with aconcentration- and treating time-dependent manner,and a dose-dependent promoting effect on SMCproliferation.These data indicated that the two risk factors for atherosclerosis had the function of inducingde novo methylation in the promoter region of the ERα gene of SMCs. However,high concentrations (50rag/L) of ox-LDL induced demethylation,indicating that different risk factors of atherosclerosis with differentpotency might cause different aberrant methylation patterns in the promoter region of the ERα gene.Theatherogenic mechanism of Hcy might involve the hypermethylation of the ERα gene,leading to the proliferationof SMCs in atherosclerotic lesions.