The cytoplasmic domain of MT1-MMP is dispensable for migration augmentation but necessary to mediate viability of MCF-7 breast cancer cells

Membrane-type-1 Matrix Metalloproteinase (MT1-MMP) is a multifunctional protease that regulates ECM degradation, proMMP-2 activation, and varied cellular processes including migration and viability. MT1-MMP is believed to be a central mediator of tumourigenesis whose role is dictated by its functionally distinct protein domains. Both the localization and signal transduction capabilities of MT1-MMP are dependent on its cytoplasmic domain, exemplifying diverse regulatory functions. To further our understanding of the multifunctional contributions of MT1-MMP to cellular processes, we overexpressed cytoplasmic domain altered constructs in MCF-7 breast cancer cells and analyzed migration and viability in 2D culture conditions, morphology in 3D Matrigel culture, and tumorigenic ability in vivo. We found that the cytoplasmic domain was not needed for MT1-MMP mediated migration promotion, but was necessary to maintain viability during serum depravation in 2D culture. Similarly, during 3D Matrigel culture the cytoplasmic domain of MT1-MMP was not needed to initiate a protrusive phenotype, but was necessary to prevent colony blebbing when cells were serum deprived. We also tested in vivo tumorigenic potential to show that cells expressing cytoplasmic domain altered constructs demonstrated a reduced ability to vascularize tumours. These results suggest that the cytoplasmic domain regulates MT1-MMP function in a manner required for cell survival, but is dispensable for cell migration.     

Differential elemental uptake in three pseudo-metallophyte C<Subscript>4</Subscript> grasses in situ in the eastern USA

Background and aims

Elemental uptake in serpentine floras in eastern North America is largely unknown. The objective of this study was to determine major and trace element concentrations in soil and leaves of three native pseudo-metallophyte C₄ grasses in situ at five sites with three very different soil types, including three serpentine sites, in eastern USA.

Methods

Pseudo-total and extractible concentrations of 15 elements were measured and correlated from the soils and leaves of three species at the five sites.

Results

Element concentrations in soils of pseudo-metallophytes varied up to five orders of magnitude. Soils from metalliferous sites exhibited higher concentrations of their characteristic elements than non-metalliferous. In metallicolous populations, elemental concentrations depended on the element. Concentrations of major elements (Ca, Mg, K) in leaves were lower than typical toxicity thresholds, whereas concentrations of Zn were higher.

Conclusions

In grasses, species can maintain relatively low metal concentrations in their leaves even when soil concentrations are richer. However, in highly Zn-contaminated soil, we found evidence of a threshold concentration above which Zn uptake increases drastically. Finally, absence of main characteristics of serpentine soil at one site indicated the importance of soil survey and restoration to maintain serpentinophytes communities and avoid soil encroachment.

     

Constituents from Scutellaria barbata Inhibiting Nitric Oxide Production in LPS‐Stimulated Microglial Cells

The arial parts of Scutellaria barbata D. Don (Lamiaceae) efficiently inhibited NO production in BV2 microglial cells, and the active constituents were further isolated based on activity‐guided isolation using silica‐gel column chromatography, RP‐C18 MPLC and prep‐HPLC. As the results, 2 flavonoids including 6‐methoxynaringenin ( 1 ) and 6‐O‐methylscutellarein ( 5 ), and 6 neo‐clerodane diterpenes such as scutebarbatine W ( 2 ), scutebatas B ( 3 ), scutebarbatine B ( 4 ), scutebarbatine A ( 6 ), 6‐O‐nicotinolylscutebarbatine G ( 7 ), and scutebarbatine X ( 8 ) were isolated. The structures of these compounds were elucidated based on NMR and MS data, and the comparison of literature values. All the compounds except compound 7 inhibited NO production efficiently with IC₅₀ values of lower than 50 μm . Particularly, compounds 1 and 8 were the most efficient with IC₅₀ values of 25.8 and 27.4 μm , respectively. This is the first report suggesting the potential of S. barbata on the reduction of neuroinflammation.     

Investigation of karyotypic composition and evolution in <Emphasis Type="Italic">Lilium</Emphasis> species belonging to the section martagon

Analyzing chromosomal traits is one of the pragmatic ways to establish evolutionary and genetic database of plants that has complicated phylogenetic system. There are some conflicts on the exact phylogeny and evolutionary pathway of Lilium, and section martagon is the most complicated part among them. In this study, chromosomal traits of martagon lily species are described. All martagon lilies were analyzed with FISH (Fluorescence in situ hybridization) technique, followed by detailed karyotyping. Each species showed 2n = 2x = 24 of chromosome complement. Size of chromosomes ranged from 451.04 to 680.06 µm. 5S and 45S ribosomal DNA, general molecular markers in modern evolutionary research were used as probe in this study. Variation in rDNA loci and chromosome translocation were observed in Lilium hansonii; the highest number of 45S rDNA loci was detected in Lilium hansonii, followed by other martagon lilies, in similar locations but with differences, and chromosome translocation was observed from one individual of Lilium hansonii. Additionally, Lilium tsingtauense from Jeju-do Island, Korea was detected with two extra chromosomes. These kind of genetic variations through karyotyping indicate ongoing genetic variations in martagon lilies. In this study, precise analysis of chromosome traits in Lilium species belonging to section martagonperformed to contribute to better comprehension of the evolutionary pathway and establishment of cytogenetic database for further plant breeding research.     

Body Mass Index Cutoffs for Underweight,Overweight, and Obesity in South Korean Schoolgirls

Objectives: To establish BMI percentiles and cutoffs for underweight, overweight, and obesity in South Korean schoolgirls. Research Methods and Procedures: A total of 1229 South Korean schoolgirls aged 8 to 18 years were randomly selected to complete a self‐administered questionnaire. BMI charts and cutoffs were constructed after analyzing data from 1107 subjects. Percentile curves were established by the modified LMS method. Results: The percentiles for underweight, overweight, and obesity corresponding to BMI of 18.5, 23.0, and 25.0 kg/m² at age 18 were the 13.0th percentile, the 77.8th percentile, and the 91.2nd percentile, respectively. The corresponding prevalences of underweight, overweight, and obesity were 12.1, 12.5, and 9.8%, respectively. Discussion: We established for the first time, to our knowledge, new BMI cutoffs for ages 8 to 18 that corresponded to BMIs of 18.5, 23.0, and 25.0 kg/m² for Asian adults designated by the International Obesity Task Force. These newly established BMI cutoffs might help to estimate the prevalence of overweight and obesity in Asian children.     

cDNA cloning and mRNA expression of LIM protein gene homologue from the silkworm, Bombyx mori

LIM protein cDNA, from Bombyx mori that contains an open reading frame of 622 bp encoding 94 amino acids, was identified and characterized. The B. mori LIM protein homologue is classified into group 2 LIM proteins that contain glycine-rich LIM domain. B. mori LIM protein mRNA is up-regulated at late embryogenesis and detected in the mid-gut of 5th instar larvae.     

Pyruvate Fermentation by Oenococcus oeni and Leuconostoc mesenteroides and Role of Pyruvate Dehydrogenase in Anaerobic Fermentation

The heterofermentative lactic acid bacteria Oenococcus oeni and Leuconostoc mesenteroides are able to grow by fermentation of pyruvate as the carbon source (2 pyruvate → 1 lactate + 1 acetate + 1 CO₂). The growth yields amount to 4.0 and 5.3 g (dry weight)/mol of pyruvate, respectively, suggesting formation of 0.5 mol ATP/mol pyruvate. Pyruvate is oxidatively decarboxylated by pyruvate dehydrogenase to acetyl coenzyme A, which is then converted to acetate, yielding 1 mol of ATP. For NADH reoxidation, one further pyruvate molecule is reduced to lactate. The enzymes of the pathway were present after growth on pyruvate, and genome analysis showed the presence of the corresponding structural genes. The bacteria contain, in addition, pyruvate oxidase activity which is induced under microoxic conditions. Other homo- or heterofermentative lactic acid bacteria showed only low pyruvate fermentation activity.