Functional analysis of the honeybee (Apis mellifera L.) salivary system using proteomics

In insects, specific proteins and physiologically active molecules whose functions are related to their lifestyles are secreted from the salivary system. To investigate proteins/molecules related to the sociality of the European honeybee (Apis mellifera L.), we performed a proteomic analysis of the honeybee salivary system. The honeybee salivary system comprises two secretory glands: the postcerebral gland (PcG) and the thoracic gland (TG), both of which are connected to a common duct that opens in the mouthpart. Although most (31 out of 35) of the major proteins identified from the PcG and TG were housekeeping proteins, the spot intensities for aldolase and acetyl-CoA acyltransferase 2 were stronger in the PcG than in the TG in the 2-dimensional gel electrophoresis. Immunoblotting confirmed that the expression of these proteins was stronger in the PcG than in the TG, whereas expression was almost not detectable in the hypopharyngeal gland (HpG), suggesting that carbohydrate metabolism is enhanced in the honeybee PcG. In addition, imaginal disc growth factor 4 (IDGF4) was synthesized in the honeybee salivary system. Immunoblotting indicated IDGF4 expression was very strong in the PcG, moderate in the TG, and very weak in the HpG. A considerable amount of IDGF4 was detected in the royal jelly, while less was detected in honey, strongly suggesting that the honeybee salivary system secretes IDGF4 into the royal jelly and honey. The secreted IDGF4 might therefore affect growth and physiology of the other colony members.     

Reversion of thyrotoxic atrial fibrillation in hypothyroid state after radioiodine treatment.

Twenty patients with thyrotoxic Basedow's disease complicated by atrial fibrillation lasting more than one month despite treatment with antithyroidal drugs were treated with radioiodine supplemented with an antithyroidal drug or inorganic iodine. We classified the 20 patients on the basis of atrial fibrillation reversion into two groups, one with reversion (group I) and the other without reversion (group II). In all 12 patients in group I, T4 and T3 decreased to hypothyroid levels in 3.2 +/- 1.3 months, and one month later all patients had their sinus rhythm restored while T4 and T3 remained below normal (2.6 +/- 1.1 micrograms/dl and 77.9 +/- 34.4 ng/dl, respectively). Although T4 and T3 also decreased within 3.5 +/- 1.8 months in all 8 patients in group II, one month later, atrial fibrillation persisted while T4 and T3 (10.4 +/- 5.3 micrograms/dl and 157.7 +/- 67.5 ng/dl, respectively) rose significantly compared to those in group I (P less than 0.001 and P less than 0.01, respectively). For reversion of atrial fibrillation it is important that the onset of hypothyroidism is rapidly induced by radioiodine and that hypothyroidism continues for at least one month.     

p600 Plays Essential Roles in Fetal Development

p600 is a multifunctional protein implicated in cytoskeletal organization, integrin-mediated survival signaling, calcium-calmodulin signaling and the N-end rule pathway of ubiquitin-proteasome-mediated proteolysis. While push, the Drosophila counterpart of p600, is dispensable for development up to adult stage, the role of p600 has not been studied during mouse development. Here we generated p600 knockout mice to investigate the in vivo functions of p600. Interestingly, we found that homozygous deletion of p600 results in lethality between embryonic days 11.5 and 13.5 with severe defects in both embryo and placenta. Since p600 is required for placental development, we performed conditional disruption of p600, which deletes selectively p600 in the embryo but not in the placenta. The conditional mutant embryos survive longer than knockout embryos but ultimately die before embryonic day 14.5. The mutant embryos display severe cardiac problems characterized by ventricular septal defects and thin ventricular walls. These anomalies are associated with reduced activation of FAK and decreased expression of MEF2, which is regulated by FAK and plays a crucial role in cardiac development. Moreover, we observed pleiotropic defects in the liver and brain. In sum, our study sheds light on the essential roles of p600 in fetal development.     

Glaziovianin A, a new isoflavone, from the leaves of Ateleia glazioviana and its cytotoxic activity against human cancer cells

A new isoflavone, named glaziovianin A (1), was isolated from the leaves of Ateleia glazioviana (Legminosae) by means of a cytotoxicity-guided fractionation procedure against HL-60 leukemia cells. The chemical structure of 1 was determined by analysis of its extensive spectroscopic data. Glaziovianin A (1) displayed differential cytotoxicities in the Japanese Foundation for Cancer Research 39 cell line panel assay. The pattern of the differential cytotoxicities of 1 was found to correlate to that shown by TZT-1027, suggesting that 1 inhibited tubulin polymerization as an action mechanism. Although 1 had little influence on microtubule networks in interphase cells, 1-treated cells showed abnormal structures with unaligned chromosomes.     

The presence of methionine-enkephalin in plasma and urine in normal human subjects and various patients

The present study was performed to investigate whether or not there were enkephalins in plasma and urine in normal subjects and in patients with various diseases. Two kinds of antisera were developed to detect M-enk and L-enk. One has specific affinity with the C-terminus of methionine-enkephalin sulfoxide (M-O-enk), the oxidized form of M-enk, and the other with the N-terminus of L-enk. M-enk-like substance (MELS) was present in blood and urine in normal subjects, but not L-enk-like substance (LELS). Plasma MELS and its urinary output averaged 38 +/- 14 pg/ml (N = 19) and 605 +/- 235 ng/day (N = 15, M. +/- S.D.), respectively. There was a significant increase in plasma MELS and its urinary output in patients with pheochromocytoma. Plasma MELS did not show any significant increase or decrease in Cushing's disease. Addison's disease, panhypopituitarism or chronic glomerulonephritis. The urinary output of MELS was significantly increased in patients with essential hypertension, renovascular hypertension and primary aldosteronism, but was decreased in central diabetes insipidus.     

A correlation between mode of growth and regional ultrastructure of the plasma membrane of Schizosaccharomyces pombe as revealed by freeze-fracturing before and after filipin treatment

The ultrastructure of the plasma membrane of Schizosaccharomyces pombe was studied by freeze-fracture using invaginations of the plasma membrane as natural markers and filipin-induced deformations as artificial markers. In accord with the mode of growth of this organism, ultrastructural aspects of the plasma membrane were related to the following ring zones: the growing pole, adjacent regions, proximal regions, the new cell pole, and the middle in dividing cells. The growing pole and adjacent regions had no or only a few invaginations. Filipin induced numerous deformations in these regions. By contrast, the proximal regions of the plasma membrane had several invaginations and resisted filipin-induced deformation. Concomitantly with commitment to cytokinesis, both the invaginations and the resistance to filipin-induced deformation disappeared in the middle. The results presented here strongly suggest the existence of two states of the plasma membrane of S. pombe, a fact which correlates well with the mode of growth of this organism.     

Sustainable fruit-body formation of edible mycorrhizal <Emphasis Type="Italic">Tricholoma</Emphasis> species for 3 years in open pot culture with pine seedling hosts

Three edible mycorrhizal mushrooms, Tricholoma portentosum, T. saponaceum, and T. terreum, that had formed ectomycorrhizas with Pinus densiflora seedlings in vitro, were maintained in open pot culture for 3 years under laboratory conditions. Tricholoma portentosum and T. saponaceum produced fruit bodies several times. For T. terreum, which produced a single fruit body in the third year, this is the first report of mushroom production under controlled conditions. Morphological observation of fruit bodies indicated that they were mature, i.e., well-organized cap, stem, and gills, and basidiospores. These results suggest that cultivation of these three edible Tricholoma mushrooms is feasible.