Structure-activity relationships of α-, β(1)-, γ-, and δ-tomatine and tomatidine against human breast (MDA-MB-231), gastric (KATO-III), and prostate (PC3) cancer cells

Partial acid hydrolysis of the tetrasaccharide (lycotetraose) side chain of the tomato glycoalkaloid α-tomatine resulted in the formation of four products with three, two, one, and zero carbohydrate side chains, which were separated by high-performance liquid chromatography (HPLC) and identified by thin-layer chromatography (TLC) and liquid chromatography ion-trap time-of-flight mass spectrometry (LCMS-IT-TOF). The inhibitory activities in terms of IC(50) values (concentration that inhibits 50% of the cells under the test conditions) of the parent compound and the hydrolysates, isolated by preparative HPLC, against normal human liver and lung cells and human breast, gastric, and prostate cancer cells indicate that (a) the removal of sugars significantly reduced the concentration-dependent cell-inhibiting effects of the test compounds, (b) PC3 prostate cancer cells were about 10 times more susceptible to inhibition by α-tomatine than the breast and gastric cancer cells or the normal cells, (c) the activity of α-tomatine against the prostate cancer cells was 200 times greater than that of the aglycone tomatidine, and (d) the activity increased as the number of sugars on the aglycone increased, but this was only statistically significant at p < 0.05 for the normal lung Hel299 cell line. The effect of the alkaloids on tumor necrosis factor α (TNF-α) was measured in RAW264.7 macrophage cells. There was a statistically significant negative correlation between the dosage of γ- and α-tomatine and the level of TNF-α. α-Tomatine was the most effective compound at reducing TNF-α. The dietary significance of the results and future research needs are discussed.     

Chronology of anthropedogenesis in the Omiya tableland,Japan, based on a 14C age profile of humic acid

Volcanic ash soils along the western edge of the Omiya tableland, Japan, are covered with thick anthropogenic soil horizons. The formation of anthropogenic soil horizons occurs because of the soil dressing practice known as “Dorotsuke,” where alluvial soil materials are deposited on fields and mixed with volcanic ash topsoil by tillage over the years. To clarify the chronology of this anthropedogenesis, carbon-14 (¹⁴C) age profiles were estimated using humic acid fractions from three pedons: an anthropogenic soil, an undressed Andosol, and a Fluvisol. Soil charcoal fragments were also dated to estimate maximum burial age. Charcoal fragments displayed vertically random age distributions, indicating that the fragments may have had multiple origins. However, the age of charcoal in the lower part of the anthropogenic soil horizons indicated that the initiation of anthropedogenesis occurred later than the late 13th century. The ¹⁴C age profile of humic acid in the Andosol exhibited little variation in age with depth in the subsoil. The ¹⁴C age profile of humic acid in the Fluvisol suggested that the humic acid fraction included allochthonous old carbon (C), although the soil itself had been formed from recent sediments. The ¹⁴C age profile of humic acid in the anthropogenic soil showed features of its two component soils. The ¹⁴C ages in the volcanic ash subsoil matched with those in the Andosol, whereas the ages increased in the anthropogenic soil horizons because of supplementation with old C from alluvial soil materials. However, the peak ¹⁴C ages occurred in the lower part of the anthropogenic horizons, whereas the middle part on the peak position displayed a gradual age-depth gradient. This feature was interpreted as a sign of ¹⁴C activity equilibrium throughout anthropedogenesis. On the basis of this postulated ¹⁴C activity equilibrium, the linear age-depth gradient at the peak position was derived from differences in burial time, and burial ages were calculated by estimating steady-state ¹⁴C. The calculated ages were lower than the charcoal ages. These age estimates suggest that anthropedogenesis was initiated in the Middle Ages and reached an intermediate stage before or during the first half of the Edo period.     

Cropland soil–plant systems control production and consumption of methane and nitrous oxide and their emissions to the atmosphere

Croplands are an important source of atmospheric methane (CH₄) and nitrous oxide (N₂O), both potent greenhouse gases. Reduction of cropland CH₄ and N₂O emissions is expected to mitigate climate change. However, large uncertainty remains in the assessment and prediction of these emissions, which prevents us from establishing appropriate mitigation options and strategies. The uncertainty is attributed mainly to the high spatiotemporal variability in emissions (e.g., emission spikes of N₂O). Understanding and quantifying how hotspots of CH₄ and N₂O production in soil and then hot moments of their emissions occur would help reduce the uncertainty. This review focuses on soil–plant systems, particularly the rhizosphere, as possible hotspots of production and consumption of CH₄ and N₂O. It is well known that the rhizosphere controls CH₄ emission strongly, though each process of production and consumption remains to be quantified. On the other hand, surprisingly little attention has been paid to N₂O, besides the fact that plant roots strongly control nitrification and denitrification. We review the current knowledge of cropland CH₄ and N₂O emissions, and conclude that soil–plant interactions strongly affect cropland emissions of both gases, in which functions of plant roots affecting biogeochemical factors (e.g., availability of oxygen, labile organic carbon and inorganic nitrogen) in the rhizosphere and phenological changes are particularly important. In relation to the status of current knowledge, we discuss future research needed.     

Effects of successive soil freeze-thaw cycles on soil microbial biomass and organic matter decomposition potential of soils

Abstract

Effects of soil freeze-thaw cycles on soil microbial biomass were examined using 8 soil samples collected from various locations, including 4 arable land sites and 2 forest sites in temperate regions and 2 arable land sites in tropical regions. The amounts of soil microbial biomass C and N, determined by the chloroform fumigation and extraction method, significantly decreased by 6 to 40% following four successive soil freeze-thaw cycles (- 13 and 4°C at 12 h-intervals) compared with the unfrozen control (kept at 4°C during the same period of time as that of the freeze-thaw cycles). In other words, it was suggested that 60 to 94% of the soil microorganisms might survive following the successive freeze-thaw cycles. Canonical correlation analysis revealed a significantly positive correlation between the rate of microbial survival and organic matter content of soil (r = 0.948*). Correlation analysis showed that the microbial survival rate was also positively correlated with the pore-space whose size ranged from 9.5 to 6.0 μm (capillary-equivalent-diameter; r = 0.995**), pH(KCI) values (r = 0.925**), EC values (r = 0.855*), and pH (H₂O) values (r = 0.778*), respectively. These results suggested that the soil physicochemical properties regulating the amount of unfrozen water in soil may affect the rate of microbial survival following the soil freeze-thaw cycles. The potential of organic matter decomposition of the soils was examined to estimate the effects of the soil freeze-thaw cycles on the soil processes associated with the soil microbial communities. The soil freeze-thaw cycles led to significant 6% increase in chitin decomposition and 7% decrease in rice straw decomposition (p < 0.05), suggesting that the partial sterilization associated with the soil freeze-thaw cycles might disturb the soil microbial functions.     

A short note on seed dispersal by colobines: the case of the proboscis monkey

Although the role of primates in seed dispersal is generally well recognized, this is not the case for colobines, which are widely distributed in Asian and African tropical forests. Colobines consume leaves, seeds and fruits, usually unripe. A group of proboscis monkeys (Colobinae, Nasalis larvatus) consisting of 1 alpha‐male, 6 adult females and several immatures, was observed from May 2005 to May 2006. A total of 400 fecal samples from focal group members covering 13 months were examined, with over 3500 h of focal observation data on the group members in a forest along the Menanggul River, Sabah, Malaysia. Intact small seeds were only found in 23 of 71 samples in Nov 2005, 15 of 38 in Dec 2005 and 5 of 21 in Mar 2006. Seeds of Ficus (all <1.5 mm in length) were found in all 3 months and seeds from Antidesma thwaitesianum (all <3 mm) and Nauclea subdita (all <2 mm) only in Nov and Dec, which was consistent with members of the study group consuming fruits of these species mostly at these times. To our knowledge, these are the first records of seeds in the fecal samples of colobines. Even if colobines pass relatively few seeds intact, their high abundance and biomass could make them quantitatively significant in seed dispersal. The potential role of colobines as seed dispersers should be considered by colobine researchers.     

Acetic acid fermentability with <Emphasis Type="Italic">Clostridium thermoaceticum</Emphasis> and <Emphasis Type="Italic">Clostridium thermocellum</Emphasis> of standard compounds found in beech wood as produced in hot-compressed water

Acetic acid fermentability of various compounds from beech wood as produced in a two-step hot-compressed water treatment was evaluated by fermentation tests using standard compounds with Clostridium thermoaceticum and Clostridium thermocellum. For cellulose- and hemicellulose-derived products, the former microorganism was found to ferment compounds with low molecular weights such as monosaccharides, decomposed products, and organic acids to acetic acid, while the latter was found to ferment compounds with high molecular weights such as polysaccharides and oligosaccharides to acetic acid. Lignin-derived products were, on the other hand, fermented by both microorganisms to acetic acid. Based on these lines of evidence, co-culture with C. thermoaceticum and C. thermocellum was evaluated and proven to increase acetic acid fermentability. Consequently, almost all compounds produced from beech wood in hot-compressed water were found to be converted to acetic acid when using these microorganisms in combination. Thus, hot-compressed water treatment coupled with acetic acid fermentation would likely be a powerful method to produce acetic acid from lignocellulosics.     

Polyphenol composition of a functional fermented tea obtained by tea-rolling processing of green tea and loquat leaves

Phenolic constituents of a new functional fermented tea produced by tea-rolling processing of a mixture (9:1) of tea leaves and loquat leaves were examined in detail. The similarity of the phenolic composition to that of black tea was indicated by high-performance liquid chromatography comparison with other tea products. Twenty-five compounds, including three new catechin oxidation products, were isolated, and the structures of the new compounds were determined to be (2R)-2-hydroxy-3-(2,4,6-trihydroxyphenyl)-1-(3,4,5-trihydroxyphenyl)-1-propanone 2-O-gallate, dehydrotheasinensin H, and acetonyl theacitrin A by spectroscopic methods. In addition, theacitrinin A and theasinensin H were obtained for the first time from commercial tea products. Isolation of these new and known compounds confirms that reactions previously demonstrated by in vitro model experiments actually occur when fresh tea leaves are mechanically distorted and bruised during the production process.     

Accumulation of nitrous oxide and depletion of oxygen in seasonally frozen soils in northern Japan - Snow cover manipulation experiments

It has been suggested that soil-thawing and snow-melting are critical triggers for vigorous emissions of nitrous oxide (N₂O) from soils in cold regions. However, because soil freezing is affected by air temperature and snow cover, accurate predictions that estimate subsequent emissions of this important greenhouse gas are difficult to make. In this study, we measured in situ soil gas N₂O and oxygen (O₂) concentrations at two experimental sites in northern Japan over the period of a year, from November 2008 to October 2009, to clarify the factors stimulating N₂O production in soil at low temperatures. The sites were N-fertilized bare arable lands with different soil frost depths and snowmelt rates, according to the snow cover management imposed. Winter-to-spring net N₂O fluxes, ranging from −0.10 to 1.95 kg N₂O-N ha⁻¹, were positively correlated with the annual maximum soil frost depth (ranging from 0.03 to 0.41 m; r = 0.951***). In the plots with deeper maximum soil frost, winter-to-spring N₂O fluxes represented 58% to 85% of the annual values. Soil N₂O production was stimulated when the soil frost depth was greater than 0.15 m or the daily mean soil temperature at 0.05-m depth was below −2.0 °C. In the soil with the greatest frost depth, soil gas N₂O concentrations at the depth of 0.10 m peaked at 46 ppm when soil gas O₂ concentrations fell down to 0.12 m³ m⁻³ under soil temperature below 0.0 °C. Snowmelt acceleration had no stimulating effect on N₂O production in the soil during the winter-to-spring period.     

A study on freeze-drying as a method of preserving mouse sperm

This review describes the study of freeze-dried mouse sperm for practical application in preserving and transporting genetic resources. Freeze-dried sperm can be used to preserve and transport genetic resources; however, there still remain many areas which need to be studied. In particular, it is essential to assure long-term preservation over several decades or centuries. Recently, the theory of accelerated degradation kinetics to freeze-dried mouse sperm has been applied, and found that long-term preservation by conventional methods requires temperatures lower than -80 C. When the relationship between the pressure at primary drying and the preservation potential of freeze-dried mouse sperm was examined, a pressure of 0.37 mbar at primary drying significantly improved the developmental rate to the blastocyst stage. In addition, it has been shown that freeze-dried sperm stored at -80 C with and without transportation can retain their ability to generate viable offspring after storage for up to 2 years. Sperm chromatin structure assay (SCSA) was applied to mouse sperm freeze-dried under several conditions and compared the results with the embryonic developmental rates of freeze-dried sperm after intracytoplasmic sperm injection (ICSI) and with comet assay results. Furthermore, SCSA might be useful for estimation of developmental potential of fertilized eggs derived from ICSI using freeze-dried sperm in mice.     

Production of stem cell factor in canine mast cell tumors

Mast cell tumor (MCT) is the most common cutaneous tumor in dogs. We recently revealed that production of stem cell factor (SCF) contributes to the proliferation of neoplastic mast cells in an autocrine/paracrine manner. The aim of the present study was to determine the contribution of the mechanism in clinical MCTs. In consequence, high SCF expression (>10 times compared to HRMC cells) was observed in 5 of 7 MCT samples used in the study regardless of KIT mutation, which was confirmed in immunohistochemical analysis. In addition, production of SCF was observed in Ki-67-positive cells in the MCT xenograft. These results indicate the broad contribution of SCF autocrine/paracrine mechanism on clinical MCTs, providing the rationale for the clinical use of KIT inhibitors regardless of KIT mutation.