Nonequilibrium as a source of unmixing

The intermediates, U and V, of a simple model reaction system (the so-called Brusselator) are shown to have both uniform and patterned steady states in a membrane. The patterned distribution of U and V has the symmetry of hexagons in the plane and is stable for B #62; B_m, B being the controlled concentration of one of the parametric species. The uniform solution is stable for B < B_c, and, since B_c #62; B_m, the increase and subsequent decrease of B over an interval that includes (B_m,B_c) can produce a hysteresis in which the transitions from mixed (i.e. uniform) to unmixed (i.e. hexagonlly patterned) states happen quite suddenly at the two critical values. Since the transfer or reactive properties of the membrane might be different in the two states, this model provides a bistable membrane element that might have prototypical value in a theory of biochemical control.     

R-stippled maize as a transposable element system

The I-R element at the R locus destabilizes kernel pigmentation giving the variegated pattern known as stippled ( R-st). In trans linkage phase with R-st the element was shown to act as a modifier of stippled, intensifying seed spotting in parallel with effects of the dominant linked modifier M-st. Presence of I-R in the genome was, therefore, shown to be detectable as a modifier of R-st. When this test was used, new modifiers resembling M-st were often detected following mutations of R-st to the stable allele R-sc. Such mutations evidently occurred by transposition of I-R away from the R locus to a site where it was identifiable as a modifier. M-st may be such a transposed I-R. Analysis of mutations to R-sc during the second (sperm-forming) mitosis in pollen grains showed that some of the transposed I-R elements were linked with R, whereas others assorted independently. Their strengths varied from barely discernible to a level equal to M-st. Overreplication frequently accompanied transposition at the sperm-forming mitosis, leading to transposed I-R elements in both the mutant and nonmutant sperm.     

Mycobacterium tuberculosis complex mycobacteria as amoeba-resistant organisms

Background

Most environmental non-tuberculous mycobacteria have been demonstrated to invade amoebal trophozoites and cysts, but such relationships are largely unknown for members of the Mycobacterium tuberculosis complex. An environmental source has been proposed for the animal Mycobacterium bovis and the human Mycobacterium canettii.

Methodology/Principal Findings

Using optic and electron microscopy and co-culture methods, we observed that 89±0.6% of M. canettii, 12.4±0.3% of M. tuberculosis, 11.7±2% of M. bovis and 11.2±0.5% of Mycobacterium avium control organisms were phagocytized by Acanthamoeba polyphaga, a ratio significantly higher for M. canettii (P = 0.03), correlating with the significantly larger size of M. canetti organisms (P = 0.035). The percentage of intraamoebal mycobacteria surviving into cytoplasmic vacuoles was 32±2% for M. canettii, 26±1% for M. tuberculosis, 28±2% for M. bovis and 36±2% for M. avium (P = 0.57). M. tuberculosis, M. bovis and M. avium mycobacteria were further entrapped within the double wall of <1% amoebal cysts, but no M. canettii organisms were observed in amoebal cysts. The number of intracystic mycobacteria was significantly (P = 10⁻⁶) higher for M. avium than for the M. tuberculosis complex, and sub-culturing intracystic mycobacteria yielded significantly more (P = 0.02) M. avium organisms (34×10⁴ CFU/mL) than M. tuberculosis (42×10¹ CFU/mL) and M. bovis (35×10¹ CFU/mL) in the presence of a washing fluid free of mycobacteria. Mycobacteria survived in the cysts for up to 18 days and cysts protected M. tuberculosis organisms against mycobactericidal 5 mg/mL streptomycin and 2.5% glutaraldehyde.

Conclusions/Significance

These data indicate that M. tuberculosis complex organisms are amoeba-resistant organisms, as previously demonstrated for non-tuberculous, environmental mycobacteria. Intercystic survival of tuberculous mycobacteria, except for M. canettii, protect them against biocides and could play a role in their life cycle.     

Flavonoids as chemotaxonomic markers for cultivated Amazonian coca

Purported ‘Amazonian coca,’ Erythroxylum coca var. ipadu Plowman (E. coca var. ipadu) was harvested from cultivated fields in Colombia, South America to determine: (a) its identity; (b) if its leaf flavonoids were complimentary to those present in leaf tissue of E. c. var. ipadu in our collection derived from Colombia; and (c) if complimentary, indicative of kinship to E. coca var. ipadu obtained from Colombia, or a related Erythroxylum taxon. Leaf extracts from Amazonian field-grown coca afforded eight O-conjugated flavonoids: two O-conjugates of taxifolin, one O-conjugate of quercetin, two O-conjugates of eriodictyol and three O-conjugates of kaempferol. Present also in leaf tissue of Amazonian field-grown coca, but lacking in leaf tissue from our collection of E. c. var. ipadu was an O-ethyl ester typically found in E. coca var. coca, kaempferols and a 7-O-rutinoside commonly encountered in the E. novogranatense taxons. Flavonoids found in our collection of E. coca var. ipadu were five O-conjugated derivatives of taxifolin and an O-conjugated quercetin. Leaf flavonoids of currently cultivated Amazonian coca are a mixture of those present in E. coca var. coca, E. coca var. ipadu and E. novogranatense var. truxillense, whereas those present in our authenticated living collection are derivatives of E. coca var. coca. Our data suggest that the Amazonian coca under cultivation in Colombia is a genetic hybrid cross between E. coca var. coca and E. novogranatense var. truxillense, occurring after 1972.     

Chikungunya Virus and Aedes Mosquitoes: Saliva Is Infectious as soon as Two Days after Oral Infection

Background

Aedes aegypti and Aedes albopictus are potential vectors of chikungunya virus (CHIKV). The recent CHIKV outbreaks were caused by a new variant characterized by a mutation in the E1 glycoprotein gene (E1-226V) which has favored a better transmissibility by Ae. albopictus. As Ae. albopictus tends to replace Ae. aegypti in many regions, one question remained: is Ae. albopictus as efficient as Ae. aegypti to transmit the variant E1-226V of CHIKV?

Methodology and Findings

We infected orally both species with the variant E1-226V and estimated the infection, the viral dissemination, and the transmission rate by real time RT-PCR. Additionally, we used an in vitro assay to determine the amount of virus delivered by mosquitoes in their saliva. We found that Ae. aegypti as well as Ae. albopictus ensured a high replication of the virus which underwent an efficient dissemination as detectable in the salivary glands at day 2 post-infection (pi). Infectious CHIKV particles were delivered by salivary glands from day 2 with a maximum at day 6 pi for Ae. albopictus (10^3.3 PFU) and day 7 pi for Ae. aegypti (10^2.5 PFU).

Conclusions

Ae. albopictus is slightly more efficient than Ae. aegypti to transmit the variant E1-226V of CHIKV. These results will help to design an efficient vector control to limit transmission as soon as the first human cases are diagnosed.     

Optimization of diarylpentadienones as chemotherapeutics for prostate cancer

Our earlier studies indicate that (1E,4E)-1,5-bis(1-alkyl-1H-imidazol-2-yl)penta-1,4-diene-3-ones and (1E,4E)-1,5-bis(1-alkyl-1H-benzo[d]imidazol-2-yl)penta-1,4-diene-3-ones exhibit up to 121-fold greater antiproliferative potency than curcumin in human prostate cancer cell models, but only 2–10 fold increase in mouse plasma concentrations. The present study aims to further optimize them as anti-prostate cancer agents with both good potency and bioavailability. (1E,4E)-1,5-Bis(1H-imidazol-2-yl)penta-1,4-diene-3-one, the potential metabolic product of (1E,4E)-1,5-bis(1-alkyl-1H-imidazol-2-yl)penta-1,4-diene-3-ones, was synthesized and evaluated for its anti-proliferative activity. The promising potency of 1,5-bis(1-alkyl-1H-imidazol-2-yl)penta-1,4-diene-3-ones was completely abolished by removing the 1-alkyl group, suggesting the critical role of an appropriate group on the N1 position. We then envisioned that N-aryl substitution to exclude the C–H bond on the carbon adjacent to the N1 position (α-H) may increase the metabolic stability. Consequently, seven (1E,4E)-1,5-bis(1-aryl-1H-imidazol-2-yl)penta-1,4-dien-3-ones and three (1E,4E)-1,5-bis(1-aryl-1H-benzo[d]imidazol-2-yl)penta-1,4-dien-3-ones, as well as three (1E,4E)-1,5-bis(1-aryl-1H-pyrrolo[3,2-b]pyridine-2-yl)penta-1,4-dien-3-ones, were synthesized through a three-step transformation, including N-arylation via Ullmann condensation, formylation, and Horner-Wadsworth-Emmons reaction. Six optimal (1E,4E)-1,5-bis(1-aryl-1H-imidazol-2-yl)penta-1,4-dien-3-ones exhibit 24- to 375-fold improved potency as compared with curcumin. Replacement of the imidazole with bulkier benzoimidazole and 4-azaindole results in a substantial decrease in the potency. (1E,4E)-1,5-Bis(1-(2-methoxyphenyl)-1H-imidazol-2-yl)penta-1,4-dien-3-one (17d) was established as an optimal compound with both superior potency and good bioavailability that is sufficient to provide the therapeutic efficacy necessary to suppress in vivo tumor growth.     

Leaf flavonoid glycosides as chemosystematic characters in Ocimum

Thirty-one accessions of nine species belonging to three subgenera of Ocimum (basil, family Lamiaceae) were surveyed for flavonoid glycosides. Substantial infraspecific differences in flavonoid profiles of the leaves were found only in O. americanum, where var. pilosum accumulated the flavone C-glycoside, vicenin-2, which only occurred in trace amounts in var. americanum and was not detected in cv. Sacred. The major flavonoids in var. americanum and cv. Sacred, and also in all other species investigated for subgenus Ocimum, were flavonol 3-O-glucosides and 3-O-rutinosides. Many species in subgenus Ocimum also produced the more unusual compound, quercetin 3-O-(6″-O-malonyl)glucoside, and small amounts of flavone O-glycosides. The level of flavonol glycosides produced was reduced significantly in glasshouse-grown plants, but levels of flavone glycosides were unaffected. A single species investigated from subgenus Nautochilus, O. lamiifolium, had a different flavonoid glycoside profile, although the major compound was also a flavonol O-glycoside. This was identified as quercetin 3-O-xylosyl(1‴→2″)galactoside, using NMR spectroscopy. The species investigated from subgenus Gymnocimum, O. tenuiflorum (=O. sanctum), was characterised by the accumulation of flavone O-glycosides. These were isolated, and identified as the 7-O-glucuronides of luteolin and apigenin. Luteolin 5-O-glucoside was found in all nine species of Ocimum studied, and is considered to be a key character for the genus.     

Transposable elements: not as quiet as a mouse

In this issue of Genome Biology, Nellåker et al. show massive purging of deleterious transposable element variants, through negative selection, in 18 mouse strains.     

Veronica: Acylated flavone glycosides as chemosystematic markers

HPLC/DAD and LC–MS of an extract of Veronica spicata (subgenus Pseudolysimachium, Plantaginaceae) revealed the presence of six 6-hydroxyluteolin glycosides acylated with phenolic acids, three of which are new compounds and which we called spicosides. A flavonoid survey of seven more species belonging to V. subgenus Pseudolysimachium and eight species of V. subgenus Pentasepalae showed that all the Pseudolysimachium species and four of the Pentasepalae species produced 6-hydroxyflavone glycosides, whereas the remaining four Pentasepalae species contained acetylated 8-hydroxyflavone glycosides instead. Spicosides appeared to be common in subgenus Pseudolysimachium (detected in five out of eight species), but we did not find them in subgenus Pentasepalae. Previously, acetylated 8-hydroxyflavone glycosides have been isolated from or detected in eight species of V. subgenus Pentasepalae (in 13 species altogether including the new results) and seven species of V. subgenus Pocilla. However, 6-hydroxyflavone glycosides occur in some other species of these subgenera. The results are discussed in an evolutionary context.     

Zeins as indicators of maize-Tripsacum introgression

A survey of zeins in tripsacoid and non-tripsacoid races of maize from Mesoamerica and from South America, annual teosinte, perennial species of Zea and species of Tripsacum revealed at least 33 zein proteins as determined by isoelectric focusing. Zea and Tripsacum and generally also species within these genera are characterized by distinct combinations of zein proteins. Maize is extensively heterogenous, and spans the complete spectrum of zeins present in wild Zea taxa. A comparison of zein proteins failed to distinguish between introgression of maize with Tripsacum or teosinte. The ease with which maize crosses naturally with wild Zea taxa, and the rarity of hybrids with Tripsacum essentially rule out natural Tripsacum introgression as a mode of racial evolution in maize.     

成都市沙河主要绿化树种固碳释氧和降温增湿效益

以成都市沙河植物廊道广泛应用的8种绿化植物为材料,利用LI-6400XT便携式光合测定系统进行了光合生理生态指标的测定,并对其固碳释氧与降温增湿效应进行了量化研究.结果表明:整个生长季节同类植物各季节的单位叶面积固碳释氧和 降温增湿能力表现出夏季＞秋季＞春季.日固碳释氧能力由强到弱为桂花、垂柳、香樟、黄葛树、山杜英、银杏、天竺桂、水杉,年固碳释氧能力由强到弱为垂柳、香樟、黄葛树、银杏、桂花、天竺桂、水杉、山杜英,日降温增湿效果由强到弱为垂柳、山杜英、水杉、天竺桂、黄葛树、香樟、银杏、桂花.据估算,整个沙河植物群落中乔木树种年总固碳量约为5.87×104 t,总释氧量约为4.27×104 t.根据对主要树种固碳释氧和降温增湿能力的分析表明,在树种配置时,垂柳、桂花、山杜英、香樟为优选乔木树种,而银杏的固碳释氧和降温增湿能力较弱,可作为长寿树种和观赏树种适量引种,不宜大面积绿化.     

Novel pyridazinone derivatives as butyrylcholinesterase inhibitors

In the current study, forty-four new [3-(2/3/4-methoxyphenyl)-6-oxopyridazin-1(6H)-yl]methyl carbamate derivatives were synthesized and evaluated for their ability to inhibit electric eel acetylcholinesterase (EeAChE) and equine butyrylcholinesterase (eqBuChE) enzymes. According to the inhibitory activity results, [3-(2-methoxyphenyl)-6-oxopyridazin-1(6H)-yl]methyl heptylcarbamate (16c, eqBuChE, IC₅₀ = 12.8 μM; EeAChE, no inhibition at 100 μM) was the most potent eqBuChE inhibitor among the synthesized compounds and was found to be a moderate inhibitor compared to donepezil (eqBuChE, IC₅₀ = 3.25 μM; EeAChE, IC₅₀ = 0.11 μM). Kinetic and molecular docking studies indicated that compounds 16c and 14c (hexylcarbamate derivative, eqBuChE, IC₅₀ = 35 μM; EeAChE, no inhibition at 100 μM) were mixed-type inhibitors which accommodated within the catalytic active site (CAS) and peripheral anionic site (PAS) of hBuChE through stable hydrogen bonding and π-π stacking. Furthermore, it was determined that [3-(2-methoxyphenyl)-6-oxopyridazin-1(6H)-yl]methyl (4-methylphenyl)carbamate 7c (eqBuChE, IC₅₀ = 34.5 μM; EeAChE, 38.9% inhibition at 100 μM) was the most active derivative against EeAChE and a competitive inhibitor binding to the CAS of hBuChE. As a result, 6-(2-methoxyphenyl)pyridazin-3(2H)-one scaffold is important for the inhibitory activity and compounds 7c, 14c and 16c might be considered as promising lead candidates for the design and development of selective BuChE inhibitors for Alzheimer’s disease treatment.     

New acrylamide-sulfisoxazole conjugates as dihydropteroate synthase inhibitors

New functionalized acrylamide derivatives bearing sulfisoxazole moiety were designed to target bacterial dihydropteroate synthase (DHPS). The in vitro antimicrobial activities of these compounds were assessed. The E-configuration of compound 5b was proved by single crystal X-ray analysis. Compounds 5g and 5h displayed double the activity of ampicillin against B. subtilis. Also, 5h was two times more active than gentamycin against E. coli. Interestingly, compounds 5f-g, 7c, 8a, 8c exhibited two folds the potency of amphotericin B against S. racemosum while 5h displayed three folds the activity of amphotericin B against S. racemosum. Most of the synthesized compounds showed superior activities to the parent sulfisoxazole and were non-toxic to normal cells. DHPS is confirmed to be a putative target for our compounds via antagonizing their antibacterial activity by the folate precursor (p-aminobenzoic acid) and product (methionine) on E. coli ATCC 25922. Docking experiments against DHPS rationalized the observed antibacterial activity. Additionally, compound 5g was evaluated as a selective targeting vector for ^99mTc that showed a remarkable uptake and targeting ability towards the infection site that was induced in mice.     

Evaluation of Parkinson Disease Risk Variants as Expression-QTLs

The recent Parkinson Disease GWAS Consortium meta-analysis and replication study reports association at several previously confirmed risk loci SNCA, MAPT, GAK/DGKQ, and HLA and identified a novel risk locus at RIT2. To further explore functional consequences of these associations, we investigated modification of gene expression in prefrontal cortex brain samples of pathologically confirmed PD cases (N = 26) and controls (N = 24) by 67 associated SNPs in these 5 loci. Association between the eSNPs and expression was evaluated using a 2-degrees of freedom test of both association and difference in association between cases and controls, adjusted for relevant covariates. SNPs at each of the 5 loci were tested for cis-acting effects on all probes within 250 kb of each locus. Trans-effects of the SNPs on the 39,122 probes passing all QC on the microarray were also examined. From the analysis of cis-acting SNP effects, several SNPs in the MAPT region show significant association to multiple nearby probes, including two strongly correlated probes targeting the gene LOC644246 and the duplicated genes LRRC37A and LRRC37A2, and a third uncorrelated probe targeting the gene DCAKD. Significant cis-associations were also observed between SNPs and two probes targeting genes in the HLA region on chromosome 6. Expanding the association study to examine trans effects revealed an additional 23 SNP-probe associations reaching statistical significance (p<2.8×10⁻⁸) including SNPs from the SNCA, MAPT and RIT2 regions. These findings provide additional context for the interpretation of PD associated SNPs identified in recent GWAS as well as potential insight into the mechanisms underlying the observed SNP associations.     

Tree leaves as complete feed for goat bucks

The objective was to look for the promising tree leaves, for feeding to livestock. The preliminary screening of tree leaves (nine species) by in vitro gas production technique revealed that leaves of Azadirachta indica, Melia azedarach, Morus alba and Leucaena leucocephala could serve as promising, alternate feed resource for ruminants. Therefore, in vivo evaluation of these tree leaves (except A. indica) along with that of Toona ciliate was assessed. Each species of the fresh tree leaves, supplemented with mineral mixture and common salt, was offered ad lib as complete feed to three bucks (Beetle × Anglo Nubian × French Alpine; 6 years old of 56.7 ± 1.12 kg BW). The data were analyzed by using completely randomized design. Bucks relished all the tree leaves (except T. ciliate) as indicated by higher (P < 0.05) voluntary DM intake and digestibility of nutrients. All the animals were in positive N-balance except those fed Toona ciliate. The N-retention was maximum in animals fed M. alba followed by that in L. leucocephala and M. azedarach. However, the DCP content of M. alba, M. azedarach and L. leucocephala was statistically comparable, while ME availability was higher (P < 0.05) from leaves of M. azedarach followed by that of M. alba, clearly indicated that leaves of M. azedarach, M. alba or L. leucocephala, supplemented with mineral mixture and common salt, could serve as an excellent complete feed for small ruminants.     

Veillonella,Firmicutes: Microbes disguised as Gram negatives

The Firmicutes represent a major component of the intestinal microflora. The intestinal Firmicutes are a large, diverse group of organisms, many of which are poorly characterized due to their anaerobic growth requirements. Although most Firmicutes are Gram positive, members of the class Negativicutes, including the genus Veillonella, stain Gram negative. Veillonella are among the most abundant organisms of the oral and intestinal microflora of animals and humans, in spite of being strict anaerobes. In this work, the genomes of 24 Negativicutes, including eight Veillonella spp., are compared to 20 other Firmicutes genomes; a further 101 prokaryotic genomes were included, covering 26 phyla. Thus a total of 145 prokaryotic genomes were analyzed by various methods to investigate the apparent conflict of the Veillonella Gram stain and their taxonomic position within the Firmicutes. Comparison of the genome sequences confirms that the Negativicutes are distantly related to Clostridium spp., based on 16S rRNA, complete genomic DNA sequences, and a consensus tree based on conserved proteins. The genus Veillonella is relatively homogeneous: inter-genus pair-wise comparison identifies at least 1,350 shared proteins, although less than half of these are found in any given Clostridium genome. Only 27 proteins are found conserved in all analyzed prokaryote genomes. Veillonella has distinct metabolic properties, and significant similarities to genomes of Proteobacteria are not detected, with the exception of a shared LPS biosynthesis pathway. The clade within the class Negativicutes to which the genus Veillonella belongs exhibits unique properties, most of which are in common with Gram-positives and some with Gram negatives. They are only distantly related to Clostridia, but are even less closely related to Gram-negative species. Though the Negativicutes stain Gram-negative and possess two membranes, the genome and proteome analysis presented here confirm their place within the (mainly) Gram positive phylum of the Firmicutes. Further studies are required to unveil the evolutionary history of the Veillonella and other Negativicutes.