Quantitative Analysis of Flavonoids in Chamomile Flowers (<Emphasis Type="Italic">Matricaria chamomilla</Emphasis> L.) by Microcolumn HPLC-UV

A reverse-phase microcolumn HPLC method with UV detection (330 nm) was developed for quantitative analysis of flavonoids of Matricaria chamomilla flowers using a ProntoSIL-120-5-C18 AQ column (60 mm × 1 mm × 5 μm), gradient elution system 0.2 M LiClO₄/0.006 M HClO₄–acetonitrile and cosmosiin as a reference compound. Optimal conditions for the hydrolytic process of flavonoids (KOH concentration 0.25%, extraction time 30 min) and the parameters of flavonoids extraction (particle size 0.25 mm, extraction temperature 60°C, a single extraction step lasting 30 min at a ratio 1: 100) were selected. Validation analysis showed that the proposed method is characterized by satisfactory metrological parameters. The limit of detection (LOD) and limit of quantification (LOQ) of cosmosiin were 84 and 255 ng/mL, respectively. The accuracy for cosmosiin content levels 80–120% was less than 101.93–103.00%. The method was used for the analysis of introduced and commercial samples of M. chamomilla flowers.     

Patterns of DNA Barcode Variation in Canadian Marine Molluscs

Background

Molluscs are the most diverse marine phylum and this high diversity has resulted in considerable taxonomic problems. Because the number of species in Canadian oceans remains uncertain, there is a need to incorporate molecular methods into species identifications. A 648 base pair segment of the cytochrome c oxidase subunit I gene has proven useful for the identification and discovery of species in many animal lineages. While the utility of DNA barcoding in molluscs has been demonstrated in other studies, this is the first effort to construct a DNA barcode registry for marine molluscs across such a large geographic area.

Methodology/Principal Findings

This study examines patterns of DNA barcode variation in 227 species of Canadian marine molluscs. Intraspecific sequence divergences ranged from 0–26.4% and a barcode gap existed for most taxa. Eleven cases of relatively deep (>2%) intraspecific divergence were detected, suggesting the possible presence of overlooked species. Structural variation was detected in COI with indels found in 37 species, mostly bivalves. Some indels were present in divergent lineages, primarily in the region of the first external loop, suggesting certain areas are hotspots for change. Lastly, mean GC content varied substantially among orders (24.5%–46.5%), and showed a significant positive correlation with nearest neighbour distances.

Conclusions/Significance

DNA barcoding is an effective tool for the identification of Canadian marine molluscs and for revealing possible cases of overlooked species. Some species with deep intraspecific divergence showed a biogeographic partition between lineages on the Atlantic, Arctic and Pacific coasts, suggesting the role of Pleistocene glaciations in the subdivision of their populations. Indels were prevalent in the barcode region of the COI gene in bivalves and gastropods. This study highlights the efficacy of DNA barcoding for providing insights into sequence variation across a broad taxonomic group on a large geographic scale.     

Galactomannan from the seeds of Ural licorice (Glycyrrhiza uralensis Fisch.)

Galactomannans from the seeds of Ural licorice (Glycyrrhiza uralensis Fisch.) obtained by hot water extraction of freshly ripened (GGu-1) and overwintered (GGu-2) seeds were studied. GGu-1 and GGu-2 (yield, 1.98 and 1.99% of the seed weight) had molecular weights of 1379 and 877 kDa, respectively; their solutions were characterized by high viscosity ([eta] 1193.1 and 765.8 mg/g, respectively) and optical activity ([alpha]D +64.8 and +65.6 deg, respectively). Their galactose-to-mannose ratio was 1 : 1.52 and 1 : 1.50, respectively. According to IR and 13C NMR spectroscopic data and methylation analysis, the polymeric chains of GGu-1 and GGu-2 are comprised of 1,4-beta-D-mannopyranose residues substituted at C-6 with single alpha-D-galactopyranose residues. The content of mannobiose units Man-Man, (Gal)Man-Man / Man-Man(Gal), and (Gal)Man-Man(Gal) differentially substituted with galactose in macromolecules GGu-1 and GGu-2 was 25.2, 18.4 and 55.9% for GGu-1 and 26.5, 32.5, and 41.0% for GGu-2.     

Expression of miRNAs in ovine fetal gonads: potential role in gonadal differentiation

Background  

Gonadal differentiation in the mammalian fetus involves a complex dose-dependent genetic network. Initiation and progression of fetal ovarian and testicular pathways are accompanied by dynamic expression patterns of thousands of genes. We postulate these expression patterns are regulated by small non-coding RNAs called microRNAs (miRNAs). The aim of this study was to identify the expression of miRNAs in mammalian fetal gonads using sheep as a model.     

Phenolic compounds of <Emphasis Type="Italic">Scutellaria baicalensis</Emphasis> Georgi

Information on the phenolic compounds from the Baikal scullap Scutellaria baicalensis Georgi, family Lamiaceae, is given. Structures of 125 compounds are presented, and information is given on their distribution in the aboveground and belowground parts of the plant and in the tissue culture of the root. There are generalized data on the use of TLC and HPLC methods for the separation of the main components of Scutellaria baicalensis.     

Water-soluble glucans from true cardamom (Elettaria cardamomum White at Maton) seeds

Water-soluble polysaccharides from seeds of true cardamom (Elettaria cardamomum White at Maton, family Zingiberaceae) have been studied. The study has shown the presence of neutral and acidic components in these polysaccharides. Three polysaccharides (380, 166, and 27 kDa) have been isolated from the neutral fraction. According to the structural analysis data, they represent α-glucans with different degrees of branching (7.1–46.1%); α-(1→4)-D-glucopyranose residues of their backbone chains are substituted at the C6 position with single α-D-glucopyranose residues. Polysaccharides with such structures have a wide range of biological activity. The presence of branched α-glucans in E. cardamomum seeds has been demonstrated.     

Plutella australiana (Lepidoptera,Plutellidae), an overlooked diamondback moth revealed?by?DNA?barcodes

The genus Plutella was thought to be represented in Australia by a single introduced species, Plutella xylostella (Linnaeus), the diamondback moth. Its status as a major pest of cruciferous crops, and the difficulty in developing control strategies has motivated broad-ranging studies on its biology. Prior genetic work has generally supported the conclusion that populations of this migratory species are connected by substantial gene flow. However, the present study reveals the presence of two genetically divergent lineages of this taxonin Australia. One shows close genetic and morphological similarity with the nearly cosmopolitan Plutella xylostella. The second lineage possesses a similar external morphology, but marked sequence divergence in the barcode region of the cytochrome c oxidase I gene, coupled with clear differences in genitalia. As a consequence, members of this lineage are described as a new species, Plutella australiana Landry & Hebert, which is broadly distributed in the eastern half of Australia.     

Synthetic osteogenic extracellular matrix formed by coated silicon dioxide nanosprings

Background

Angiogenesis is widely investigated in conjunction with cancer development, in particular because of the possibility of early stage detection and of new therapeutic strategies. However, such studies are negatively affected by the limitations of imaging techniques in the detection of microscopic blood vessels (diameter 3-5 ??m) grown under angiogenic stress. We report that synchrotron-based X-ray imaging techniques with very high spatial resolution can overcome this obstacle, provided that suitable contrast agents are used.

Results

We tested different contrast agents based on gold nanoparticles (AuNPs) for the detection of cancer-related angiogenesis by synchrotron microradiology, microtomography and high resolution X-ray microscopy. Among them only bare-AuNPs in conjunction with heparin injection provided sufficient contrast to allow in vivo detection of small capillary species (the smallest measured lumen diameters were 3-5 ??m). The detected vessel density was 3-7 times higher than with other nanoparticles. We also found that bare-AuNPs with heparin allows detecting symptoms of local extravascular nanoparticle diffusion in tumor areas where capillary leakage appeared.

Conclusions

Although high-Z AuNPs are natural candidates as radiology contrast agents, their success is not guaranteed, in particular when targeting very small blood vessels in tumor-related angiography. We found that AuNPs injected with heparin produced the contrast level needed to reveal--for the first time by X-ray imaging--tumor microvessels with 3-5 ??m diameter as well as extravascular diffusion due to basal membrane defenestration. These results open the interesting possibility of functional imaging of the tumor microvasculature, of its development and organization, as well as of the effects of anti-angiogenic drugs.     

Towards a comprehensive barcode library for arctic life - Ephemeroptera, Plecoptera, and Trichoptera of Churchill, Manitoba, Canada

Background

In order to understand the role of herbivores in trophic webs, it is essential to know what they feed on. Diet analysis is, however, a challenge in many small herbivores with a secretive life style. In this paper, we compare novel (high-throughput pyrosequencing) DNA barcoding technology for plant mixture with traditional microhistological method. We analysed stomach contents of two ecologically important subarctic vole species, Microtus oeconomus and Myodes rufocanus, with the two methods. DNA barcoding was conducted using the P6-loop of the chloroplast trnL (UAA) intron.

Results

Although the identified plant taxa in the diets matched relatively well between the two methods, DNA barcoding gave by far taxonomically more detailed results. Quantitative comparison of results was difficult, mainly due to low taxonomic resolution of the microhistological method, which also in part explained discrepancies between the methods. Other discrepancies were likely due to biases mostly in the microhistological analysis.

Conclusion

We conclude that DNA barcoding opens up for new possibilities in the study of plant-herbivore interactions, giving a detailed and relatively unbiased picture of food utilization of herbivores.     

Molecular evolution of mitochondrial 12S RNA and cytochrome b sequences in the pantherine lineage of Felidae

DNA sequence comparisons of two mitochondrial DNA genes were used to infer phylogenetic relationships among 17 Felidae species, notably 15 in the previously described pantherine lineage. The polymerase chain reaction (PCR) was used to generate sequences of 358 base pairs of the mitochondrial 12S RNA gene and 289 base pairs of the cytochrome b protein coding gene. DNA sequences were compared within and between 17 felid and five nonfelid carnivore species. Evolutionary trees were constructed using phenetic, cladistic, and maximum likelihood algorithms. The combined results suggested several phylogenetic relationships including (1) the recognition of a recently evolved monophyletic genus Panthera consisting of Panthera leo, P. pardus, P. onca, P. uncia, P. tigris, and Neofelis nebulosa; (2) the recent common ancestry of Acinonyx jubatus, the African cheetah, and Puma concolor, the American puma; and (3) two golden cat species, Profelis temmincki and Profelis aurata, are not sister species, and the latter is strongly associated with Caracal caracal. These data add to the growing database of vertebrate mtDNA sequences and, given the relatively recent divergence among the felids represented here (1-10 Myr), allow 12S and cytochrome b sequence evolution to be addressed over a time scale different from those addressed in most work on vertebrate mtDNA.