Characterization of the plastid genome of Cratoxylum species (Hypericaceae) and new insights into phylogenetic relationships.

To expand the genomic information of Hypericaceae, particularly on Cratoxylum, we characterized seven novel complete plastid genomes (plastomes) of five Cratoxylum and two of its allied taxa, including C. arborescens, C. formosum subsp. formosum, C. formosum subsp. pruniflorum, C. maingayi, C. sumatranum, Hypericum hookerianum, and Triadenum breviflorum. For Cratoxylum, the plastomes ranged from 156,962 to 157,792 bp in length. Genomic structure and gene contents were observed in the five plastomes, and were comprised of 128-129 genes, which includes 83-84 protein-coding (CDS), 37 tRNA, and eight rRNA genes. The plastomes of H. hookerianum and T. breviflorum were 138,260 bp and 167,693 bp, respectively. A total of 110 and 127 genes included 72 and 82 CDS, 34 and 37 tRNA, as well as four and eight rRNA genes. The reconstruction of the phylogenetic trees using maximum likelihood (ML) and Bayesian inference (BI) trees based on the concatenated CDS and internal transcribed spacer (ITS) sequences that were analyzed separately have revealed the same topology structure at genus level; Cratoxylum is monophyletic. However, C. formosum subsp. pruniflorum was not clustered together with its origin, raising doubt that it should be treated as a distinct species, C. pruniflorum based on molecular evidence that was supported by morphological descriptions.

Investigation of Morinda citrifolia Activities through Pinoresinol and α-EG Related Gene Expression.

α-EG is a unique substance that was first found in the leaves and fruits of Morinda citrifolia (Mc) growing in Thailand using GC-MS at 52.33% and 54.12%. It was then concentrated and its abundance quantified, along with that of pinoresinol, via GC, compared to the standards in leaves, ufp, rfp, rawfs, and seeds. α-EG and pinoresinol, which have collagen stimulating, skin whitening, and an inhibitory effect on wrinkle formation, were found in different concentrations and amounts. Three different concentrations of the five Mc part extracts were tested on NHDF for gene expression related to the aforementioned activities, COL1A1, COL1A2, and COL3A1, FGF1 and FGF7 by qRT-PCR. The results showed various expression levels, both stimulatory and inhibitory, with different concentrations of plant parts and genes. Similar results were revealed when the experiments were performed with Morus alba (Ma), which was found to contain 20.48 g protein p/100 g leaves at concentrations of 3.11 mg/mL. The studied Mc parts seem to have advantages based on the stated objectives, gene type and level of activity of each plant part. Rawfs and leaves supplemented with Ma samples were selected for toxicity tests with PBMCs. The lack of both cell and DNA toxicity from the rawfs indicated that they can be used safely.

Formulations for Effective Detoxification Derived from Three Medicinal Plants: Thunbergia laurifolia, Clerodendrum disparifolium and Rotheca serrata.

BACKGROUND: Based on the long history of the medicinal use of Thunbergia laurifolia, Clerodendrum disparifolium and Rotheca serrata, the extract formulations of these species: T. laurifolia and C. disparifolium; T. laurifolia and R. serrata; and T. laurifolia, C. disparifolium and R. serrata, called formulas 1, 2 and 3, were created for detoxification testing to take more advantage of each species. OBJECTIVE: The objective of this study is to estimate the detoxifying effects of studied extract formulations on human cell and tissue culture as a preclinical trial. METHODS: The major phytochemicals were derived by GC-MS. The detoxification efficacy of these formulations in cells and DNA levels were derived by MTT and comet assays in toxic PBMCs (incubated with rice whisky or bathroom cleaner). RESULTS: The phytochemical constituents were detected at 23.48% phytol and 43.03% oleamide in T. laurifolia; 12.88% oleamide, 20.93% 9,12,15-octadecatrien, 25.52% squalene, 22.19% butylated hydroxy toluene and 15.36% vitamin E in C. disparifolium; and 30.41% phytol, 32.78% oleamide, and 12.20%, 9,12,15-octadecatrien-1-ol in R. serrata. The toxic cells treated with the plant formulas 1, 2 and 3 showed no IC50 values, but formulas 1 and 2 displayed higher efficacies than formula 3 did. The comet assay indicated that the experiments (the treatment on toxic cells with the plant formulas) induced significant (p < 0.05) DNA damage compared to the negative control due to poisoning occurring before administration of the plant formulas. The OTM of the controls was significantly (p < 0.05) longer than the experimental samples showing significantly reduce the toxicity of the created formulations. CONCLUSION: The formulas showed high detoxification efficacies and formulations 1 and 2 resulted in higher levels of detoxification than formulation 3, especially in immediate treatment after receiving toxic substances.

The Unique Substance, Lidocaine and Biological Activity of the Dioscorea Species for Potential Application as a Cancer Treatment, Natural Pesticide and Product.

The six Dioscorea species, D. brevipetiolata, D. bulbifera, D. depauperata (Dd), D. glabra (Dg), D. pyrifolia and D. hamiltonii were analyzed for phytochemicals, toxicity in PBMCs, and biological activity in two cancer cell lines by MTT and comet assays, and pesticide efficiency. Via GC-MS, lidocaine was found to be the predominant compound in two of the studied species. To confirm the systematics, lidocaine was also found in lower amounts in 11 species. The MTT assay showed no toxicity in all six of the studied species. The comet assay showed the key result that the ethanol extracts of Dd and Dg violently broke DNA into pieces. Biological activity of these two species' extracts showed toxicity on HepG2 and no effects on HCT-116. The water extracts of Dd and Dg, applied to Brassica chinensis showed high efficiency as a bioprotectant. In summary, lidocaine seems to be the predominant identifying compound of the genus Dioscorea in Thailand, which is useful in systematics. At least the two species, Dd and Dg, may be used for human hepatocyte cancer treatment and as an alternative pesticide for economically important vegetables. Dioscorea species containing lidocaine or extracted lidocaine have promise for natural product creation.

First detailed reconstruction of the karyotype of Trachypithecus cristatus (Mammalia: Cercopithecidae).

BACKGROUND: The chromosomal homologies of human (Homo sapiens = HSA) and silvered leaf monkey (Trachypithecus cristatus = TCR) have been previously studied by classical chromosome staining and by fluorescence in situ hybridization (FISH) applying chromosome-specific DNA probes of all human chromosomes in the 1980s and 1990s, respectively. RESULTS: However, as the resolution of these techniques is limited we used multicolor banding (MCB) at an ~250-band level, and other selected human DNA probes to establish a detailed chromosomal map of TCR. Therefore it was possible to precisely determine evolutionary conserved breakpoints, orientation of segments and distribution of specific regions in TCR compared to HSA. Overall, 69 evolutionary conserved breakpoints including chromosomal segments, which failed to be resolved in previous reports, were exactly identified and characterized. CONCLUSIONS: This work also represents the first molecular cytogenetic one characterizing a multiple sex chromosome system with a male karyotype 44,XY1Y2. The obtained results are compared to other available data for old world monkeys and drawbacks in hominoid evolution are discussed.

Generation of multicolor banding probes for chromosomes of different species.

BACKGROUND: The multicolor banding (MCB/mBAND) technique provides a unique opportunity to characterize intrachromosomal rearrangements and to determine chromosomal breakpoints. Until recently, MCB probes have only been available for human and some murine chromosomes. Generation of MCB probes for chromosomes of other species, useful and required in many cytogenetics research fields, was limited by technical difficulties. MCB probes are established by chromosome microdissection followed by whole genomic DNA amplification. However, unambiguous identification of the target chromosome is required for MCB-probe establishment. Previously proposed protocols suggested G-banding staining or preliminary FISH with whole chromosome paints (WCP) as methods to identify the chromosome of interest. RESULTS: Here we present a complete workflow for MCB probe generation for those cases and species where chromosome morphology is too challenging to recognize target chromosomes by conventional methods and where WCP probes are not available. The workflow was successfully applied for murine chromosomes that are difficult to identify unambiguously. Additionally, we showed that glass-needle based microdissection enables establishment of a whole set of WCP paints by microdissection of individual chromosomes of a single metaphase CONCLUSIONS: The present method can be applied for generation of whole or region-specific DNA probes for species, where karyotyping of G-banded chromosomes is challenging due to similar chromosome morphology and/or chromosome banding patterns.