Cancer-preventive peptide lunasin from Solanum nigrum L. inhibits acetylation of core histones H3 and H4 and phosphorylation of retinoblastoma protein (Rb)

Lunasin, a unique 43 amino acid, 4.8 kDa cancer-chemopreventive peptide initially reported in soybean and now found in barley and wheat, has been shown to be cancer-chemopreventive in mammalian cells and in a skin cancer mouse model against oncogenes and chemical carcinogens. To identify bioactive components in traditional herbal medicines and in search for new sources of lunasin, we report here the properties of lunasin from Solanum nigrum L. (SNL), a plant indigenous to northeast Asia. Lunasin was screened in the crude extracts of five varieties of the medicinal plants of Solanaceae origin and seven other major herbal plants. An in vitro digestion stability assay for measuring bioavailability was carried out on SNL crude protein and autoclaved SNL using pepsin and pancreatin. A nonradioactive histone acetyltransferase (HAT) assay and HAT activity colorimetric assay were used to measure the inhibition of core histone acetylation. The inhibitory effect of lunasin on the phosphorylation of retinoblastoma protein (Rb) was determined by immunoblotting against phospho-Rb. Lunasin isolated from autoclaved SNL inhibited core histone H3 and H4 acetylation, the activities of the HATs, and the phosphorylation of the Rb protein. Lunasin in the crude protein and in the autoclaved crude protein was very stable to pepsin and pancreatin in vitro digestion, while the synthetic pure lunasin was digested at 2 min after the reaction. We conclude that lunasin is a bioactive and bioavailable component in SNL and that consumption of SNL may play an important role in cancer prevention.     

Effects of phytic acid on peanut allergens and allergenic properties of extracts

Phytic acid would form soluble and insoluble complexes with proteins. Our objective was to determine if phytic acid forms insoluble complexes with major peanut allergens, and if such reaction results in a peanut extract with a lower level of soluble allergens and allergenic property. Extracts from raw and roasted peanuts were treated with and without phytic acid at various pH values and then analyzed by SDS-PAGE and a competitive inhibition ELISA (ciELISA). The ciELISA measured IgE binding using a pooled serum from peanut-allergic individuals. Results showed that phytic acid formed complexes with the major peanut allergens (Ara h 1 and Ara h 2), which were insoluble in acidic and neutral conditions. Succinylation of the allergens inhibited complex formation, indicating that lysine residues were involved. A 6-fold reduction in IgE binding or allergenic potency of the extract was observed after treatment with phytic acid. It was concluded that phytic acid formed insoluble complexes with the major peanut allergens, and resulted in a peanut extract with reduced allergenic potency. Application of phytic acid to a peanut butter slurry presented a similar result, indicating that phytic acid may find use in the development of hypoallergenic peanut-based products.     

Corticotropin-releasing factor immunoreactivity increases in the cerebellar climbing fibers in the novel ataxic mutant mouse, pogo

The ataxic pogo mouse (pogo/pogo) is a novel neurological mutant, which was derived as an inbred strain (KJR/MsKist) from a Korean wild mouse. The pathological manifestations include a difficulty in maintaining a normal posture, the failure of inter-limb coordination and an inability to walk straight. In this study, we examined the distribution of corticotropin-releasing factor (CRF) immunoreactive cerebellar climbing fibres and their projections to tyrosine hydroxylase (TH) immunoreactive Purkinje cells in the cerebellum of the pogo mutant mouse using immunohistochemistry. In the pogo/pogo mouse, a subset of climbing fibres was stained more intensely for CRF than in the control. Moreover, ataxic pogo mouse, neurons of the inferior olivary nucleus projecting climbing fibres were also more intensely stained for CRF than in the control. In the pogo/pogo mouse, TH immunoreactivity was located in the Purkinje cells, whereas no TH expression was found in the control. Double immunostaining for CRF and TH in the pogo/pogo cerebellum revealed that the distribution of TH-immunoreactive Purkinje cells corresponded to terminal fields of CRF-immunoreactive climbing fibres but not to the CRF-immunoreactive mossy fibres. Therefore, we suggest that an increase of CRF level may alter the function of targeted Purkinje cells and that it is related to the ataxic phenotype in the pogo mutant mouse.     

Seasonal variation in photosystem II efficiency and photochemical reflectance index of evergreen trees and perennial grasses growing at low and high elevations in subtropical Taiwan

Three pines species, three evergreen broadleaf trees, one C(3) and two C(4) perennial grasses of subtropical Taiwan were studied to elucidate the correlation between photosystem II (PSII) efficiency and photochemical reflectance index (PRI = (R(531) - R(570))/(R(531) + R(570))). Measurements were made at two sites differing in altitude (800 and 2600 m) over several growing seasons. At high elevation, potential PSII efficiency, measured by chlorophyll fluorescence (the ratio of variable to maximal fluorescence; F(v)/F(m)) at predawn, decreased with decreasing air temperature and varied greatly among species. At the lowest air temperature (-3 degrees C) studied, variation in F(v)/F(m) among species ranged from 0.33 to 0.72. In contrast, at low elevation where air temperature was moderate, seasonal variation in F(v)/F(m) was small in all of the study species. When species, elevation and season data were pooled, despite the high variation in F(v)/F(m) among species, a good correlation between F(v)/F(m) and PRI was observed. When compared at the same value of PRI, F(v)/F(m) of evergreen trees was higher than that of perennial grasses; however, when the minimum temperature on the measurement day was below 0 degrees C, F(v)/F(m) was underestimated relative to PRI. We conclude that PRI could be used as a remote indicator of photosynthetic function when air temperature is above 0 degrees C.     

Effects of fermented mushroom (Flammulina velutipes) by‐product diets on growth performance and carcass traits in growing‐fattening Berkshire pigs

This study was conducted to investigate effects of fermented mushroom (Flammulina velutipes) by‐product diets on the growth performance and carcass traits in growing‐fattening Berkshire pigs. The fermented diets mainly contained 40.0% mushroom by‐product, 20.0% formula feed, 26.0% rice bran and supplemental 0.1% probiotics. The mixed ingredients were fermented for 5 days at room temperature. Berkshire pigs (n = 225) were divided into five groups and three replications. The basal diets (C) were substituted by 10% (T1), 30% (T2), 50% (T3) and 70% (T4) fermented mushroom by‐product diets. Crude protein concentration and total calorie in fermented diets were significantly increased (P < 0.05) at the end of fermentation days compared with initial fermentation day. Body weight gain, feed efficiency and carcass weight were significantly lower (P < 0.05) in the T2, T3 and T4 groups than in the control group. Carcass grade was significantly better (P < 0.05) in the pigs fed fermented diets than in the pigs fed control diet and the ratio of high grade (1 plus 2 grades) was higher in the fermented diet groups compared with the control group. Therefore, although a diet of fermented mushroom by‐product decreased growth performance and feed efficiency, it improved the carcass grade in Berkshire pigs.     

Infection of normal C3H/HeN mice with Taenia saginata asiatica oncospheres

Normal C3H/HeN female mice were used to develop an animal model of Taenia saginata asiatica oncosphere infection. The host cellular immune response in this model was analyzed by a cytokine enzyme-linked immunosorbent assay (cytokine ELISA) and flow cytometry. Tumor-like cysts containing cysticerci were recovered from the inoculation sites of female mice 7 weeks postinfection with the T. saginata asiatica oncospheres. A sharp increase and sustained elevation in the ability of spleen cells to produce interferon-γ and interleukin (IL)-2 revealed that cellular immunity played an important role during the infection. An immediate increase in the levels of IL-6 at 1 week postinfection indicated the induction of a local acute inflammatory response. However, no significant change in the levels of IL-10 indicated that Th2 cells were not involved in this immune response. The patterns of cell distribution revealed by flow cytometry also supported the same finding. These results suggested that Th1 cells played a major role in the immune response in C3H/HeN mice during the early stages of the oncosphere infection and that the Th2 response was not induced during the stage of cysticercus formation.     

Low dietary inorganic phosphate affects the lung growth of developing mice

Inorganic phosphate (Pi) plays a critical role in diverse cellular functions, and regulating the Pi balance is accomplished by sodium-dependent Pi co-transporter (NPT). Pulmonary NPT has recently been identified in mammalian lungs. However, to date, many of the studies that have involved Pi have mainly focused on its effect on bone and kidney. Therefore, current study was performed to discover the potential effects of low Pi on the lung of developing transgenic mice expressing the renilla/firefly luciferase dual reporter gene. Two-weeks old male mice divided into 2 groups and these groups were fed either a low PI diet or a normal control diet (normal: 0.5% Pi, low: 0.1% Pi) for 4 weeks. After 4 weeks of the diet, all the mice were sacrificed. Their lungs were harvested and analyzed by performing luciferase assay, Western blotting, kinase assay and immunohistochemistry. Our results demonstrate that low Pi affects the lungs of developing mice by disturbing protein translation, the cell cycle and the expression of fibroblast growth factor-2. These results suggest that optimally regulating Pi consumption may be important to maintain health.