Fucose-containing sulfated polysaccharides from brown seaweeds inhibit proliferation of melanoma cells and induce apoptosis by activation of caspase-3 in vitro

Fucose-containing sulfated polysaccharides (FCSPs) extracted from seaweeds, especially brown macro-algae, are known to possess essential bioactive properties, notably growth inhibitory effects on tumor cells. In this work, we conducted a series of in vitro studies to examine the influence of FCSPs products from Sargassumhenslowianum C. Agardh (FSAR) and Fucus vesiculosus (FVES), respectively, on proliferation of melanoma B16 cells and to investigate the underlying apoptosis promoting mechanisms. Cell viability analysis showed that both FCSPs products, i.e., FSAR and FVES, decreased the proliferation of the melanoma cells in a dose-response fashion, with FSAR being more potent at lower dosages, and FVES being relatively more anti-proliferative than FSAR at higher dosages. Flow cytometric analysis by Annexin V staining of the melanoma cells exposed to the FCSPs products confirmed that both FSAR and FVES induced apoptosis. The FCSPs-induced apoptosis was evidenced by loss of plasma membrane asymmetry and translocation of the cell membrane phospholipids and was accompanied by the activation of caspase-3. The FCSPs bioactivity is proposed to be attributable to distinct structural features of the FCSPs, particularly the presence of sulfated galactofucans (notably in S.henslowianum) and sulfated fucans (notably in F. vesiculosus). This study thus indicates that unfractionated FCSPs may exert bioactive effects on skin cancer cells via induction of apoptosis through cascades of reactions that involve activation of caspase-3.     

Dynorphin-Kappa Opioid Receptor Signaling Partly Mediates Estrogen Negative Feedback Effect on LH Pulses in Female Rats

Accumulating evidence suggests that the arcuate nucleus (ARC) kisspeptin/neurokinin B (NKB)/dynorphin (KNDy) neurons play a role in estrogen negative feedback action on pulsatile gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) release. The present study aimed to determine if dynorphin (Dyn) is involved in estrogen negative feedback on pulsatile GnRH/LH release. The effect of the injection of nor-binaltorphimine (nor-BNI), a kappa-opioid receptor (KOR) antagonist, into the third cerebroventricle (3V) on LH pulses was determined in ovariectomized (OVX) adult female rats with/without replacement of negative feedback levels of estradiol (low E₂). The mean LH concentrations and baseline levels of LH secretion in nor-BNI-injected, low E₂-treated rats were significantly higher compared with vehicle-treated controls. On the other hand, the nor-BNI treatment failed to affect any LH pulse parameters in OVX rats without low E₂ treatment. These results suggest that Dyn is involved in the estrogen negative feedback regulation of pulsatile GnRH/LH release. The low E₂ treatment had no significant effect on the numbers of ARC Pdyn (Dyn gene)-,Kiss1- and Tac2 (NKB gene)-expressing cells. The treatment also did not affect mRNA levels of Pdyn and Oprk1 (KOR gene) in the ARC-median eminence region, but significantly increased the ARC kisspeptin immunoreactivity. These findings suggest that the negative feedback level of estrogen suppresses kisspeptin release from the ARC KNDy neurons through an unknown mechanism without affecting the Dyn and KOR expressions in the ARC. Taken together, the present result suggests that Dyn-KOR signaling is a part of estrogen negative feedback action on GnRH/LH pulses by reducing the kisspeptin release in female rats.     

Evaluation of dry matter production and yield in early-sown wheat using near-isogenic lines for the vernalization locus Vrn-D1

ABSTRACT

Wheat (Triticum aestivum L.) grain yield is predicted to decrease in the future because of an increase in air temperature globally. To clarify the effects of the vernalization response gene in wheat to warmer winters, we compared dry matter production and grain yield between spring wheat ‘Asakazekomugi’ and its winter-type near-isogenic line (NIL) carrying different alleles of the vernalization response gene Vrn-D1 under early-, standard-, and late-sowing conditions. Under early-sowing conditions, dry matter production of the NIL carrying the winter allele of Vrn-D1, named Asa (Vrn-D1b), exceeded that of ‘Asakazekomugi’ from mid-March (after stem elongation in Asa (Vrn-D1b)) when the temperatures rose. Tiller number and leaf area index under early-sowing conditions were consistently higher in Asa (Vrn-D1b) than in ‘Asakazekomugi’ from mid-March onward. It was suggested that the early-sown ‘Asakazekomugi’ could not effectively absorb solar radiation to produce dry matter because of the acceleration of stem elongation caused by the Vrn-D1 gene during the cold season. The grain yield of Asa (Vrn-D1b) with early sowing was higher than with standard sowing. In contrast, the grain yield of ‘Asakazekomugi’ was lower in the early-sown crop than in the crop sown at the standard date. These results suggested that the higher grain yield of Asa (Vrn-D1b) than that of ‘Asakazekomugi’ under early-sown conditions could be due to Asa (Vrn-D1b) maintaining high dry matter production after the jointing stage by suppressing acceleration of growth caused by warm conditions after sowing.     

Regulation of stony hard peach softening with ACC treatment

Controlling the rate of fruit softening in melting-flesh peaches is a primary goal of the fruit industry. Stony hard (SH) peach varieties lack the ability to synthesize 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene, which is required for fruit maturation. SH peaches thus have crisp flesh that remains firm during ripening. In this study, we developed a simple technique to stimulate fruit softening by a single spray application of ACC at a concentration of 10–20 mM, which was sufficient to allow ethylene synthesis and fruit softening. Higher concentrations of ACC increased ethylene production, and made the fruit softer. Ethylene synthesis was limited to the first 2–3 d after ACC treatment, after which fruit ceased softening and retained its remaining firmness. These results indicate that a single application of ACC solution can be used to regulate the process of fruit softening in SH peaches.     

Supplementing transglucosidase with a high-fiber diet for prevention of postprandial hyperglycemia in streptozotocin-induced diabetic dogs

Indigestible oligosaccharides have been shown to normalize blood glucose and insulin concentration thereby promoting good health and preventing diseases, such as diabetes. Transglucosidase (TG, α-glucosidase, enzyme code (EC) 3.2.1.20) is an enzyme capable of converting starch to oligosaccharides, such as iso-malto-oligosaccharides from maltose, via the action of amylase. The aim of this study was to evaluate whether oral administration of TG with maltose or dextrin is capable of reducing post-prandial serum glucose concentration in experimentally streptozotocin (STZ)-induced diabetic dogs fed on a high-fiber diet. Five healthy and five STZ-induced diabetic dogs were employed in this study. TG supplementation with dextrin or maltose had no detrimental effect in healthy dogs. In fact, TG and dextrin exhibited a flatlined serum glucose pattern, while reducing mean post-prandial serum insulin and glucose concentration as compared to control diet alone. When TG supplementation was tested in STZ-induced diabetic dogs under the context of a high fiber diet, a 13.8% and 23.9% reduction in mean glucose concentration for TG with maltose and dextrin, respectively was observed. Moreover, TG with dextrin resulted in a 13% lower mean post-prandial glucose concentration than TG with maltose, suggesting that dextrin may be a more efficient substrate than maltose when used at the same concentration (1 g/kg). Our results indicate that TG supplementation with diet can lead to lower postprandial glucose levels versus diet alone. However, the efficacy of TG supplementation may depend on the type of diet it is supplemented with. As such, TG administration may be useful for preventing the progression of diabetes mellitus and in its management in dogs.     

Prominent expression of spinocerebellar ataxia type-1 (SCA1) gene encoding ataxin-1 in LH-producing cells, LbetaT2

The LH-producing cell line, LbetaT2, and non LH-producing cell line, alphaT3-1 cells, established from a pituitary tumor, were employed for cDNA subtraction cloning to identify genes with expression unique to LH producing cells. Several cDNAs that code for known as well as for many unidentified clones were discovered. Most clones were the spinocerebellar ataxia type-1 (SCA1) gene encoding ataxin-1, the abnormality of which causes neurodegeneration and loss of cerebellar Purkinje cells. We examined whether the expression of SCA1 gene in LbetaT2 cells is related to hormone production. We also compared the expression of SCA1 with that in various other pituitary tumor derived cell lines, and confirmed the prominent expression of SCA1 in LbetaT2 cells. The effect of gonadal factor(s) for SCA1 gene expression was examined. The expression level in female rats was low and did not change during the estrus cycle, but increased significantly after ovariectomy and did not return to the normal level under low and high doses of estrogen. In the male pituitary SCA1 gene expression increased markedly after castration and was not decreased by estrogen or testosterone. The Ontogeny of SCA1 gene expression was investigated in porcine fetal and postnatal pituitaries and revealed biphasic and sexually dimorphic expression. Transient expression of SCA1 gene was observed at fetal day 50 and 65 in males and day 40 in females, followed by a decline and increased expression before birth in both genders. Thus the expression of SCA1 gene is prominent in LH-producing cells and is not under direct control of gonadal factor(s) in both genders. In addition to the variable expression of SCA1 gene during the fetus period, the present results provide a novel aspect to the understanding of Boucher-Neuhauser syndrome (Ataxia Hypogonadism Choroidal Dystrophy).     

Perirenal pyelocaliceal diverticulum in an infant cat

A 3-month-old female tortoise-shell cat showing azotemia died with a marked swollen abdomen. Necropsy revealed a huge perirenal cyst (8.5 x 6.0 x 4.5 cm) on the ventral aspect of the right kidney. The cyst was filled with the pellucid yellow fluid with a smell of urine. The lumen was connected with irregularly dilated renal pelvis by a narrow channel passing through the renal parenchyma. The cyst was lined by epithelial cells and its wall was consisted of collagen fibers and smooth muscle cells as that of the renal pelvis and ureter. Renal parenchyma adjacent to the channel showed interstitial infiltration of the lymphoid cells. The cyst was a diverticulum of the renal pelvis due to an impaired development.