Isolation of genes showing increased expression during bovine adipocyte differentiation

The adipocyte is important not only for the storage of excess energy as fat, but also for the secretion of homeostatic factors. Gene expression profiles during adipocyte differentiation have been reported previously for mouse 3T3‐L1 cells. However, the profiles of adipogenic gene expression in mice and cattle may be different because several metabolic pathways of the ruminant adipose tissue are different from those of non‐ruminants. The gene expression profile in a clonal bovine intramuscular preadipocyte cell line during adipogenesis was examined using the polymerase chain reaction‐subtraction method. Six hundred and twenty‐one clones, which were expressed at an early stage of differentiation, from the preadipocyte to adipocyte, were isolated and characterized. Further detailed studies were carried out for 86 selected genes using northern blotting. Ten genes were found to be highly expressed after differentiation of bovine intramuscular preadipocyte cells. In particular, the expression profiles of genes for stearoyl CoA desaturase and FK506 binding protein were quite different from the time course of differentiation of that seen in the 3T3‐L1 cells reported previously. In addition, these genes were assigned to bovine chromosomes using a bovine/hamster somatic cell hybrid panel and public database.     

Acute Phase Pulmonary Responses to a Single Intratracheal Spray Instillation of Magnetite (Fe3O4) Nanoparticles in Fischer 344 Rats

Iron nanomaterials are of considerable interest for application to nanotechnology-related fields including environmental catalysis, biomedical imaging, drug delivery and hyperthermia, because of their superparamagnetic characteristics and high catalytic abilities. However, information about potential risks of iron nanomaterials is limited. The present study assessed pulmonary responses to a single intratracheal spray instillation of triiron tetraoxide nanoparticles (magnetite) in rats. Ten-week-old male and female Fischer 344 rats (n=5/group) were exposed to a single intratracheal spray instillation of 0 (vehicle), 5.0, 15.0 or 45.0 mg/kg body weight (BW) of magnetite. After 14 days, the rats were sacrificed, and biological consequences were investigated. The lung weights of the 15.0 and 45.0 mg/kg BW male and female groups were significantly higher than those of the control groups. The lungs of treated rats showed enlargement and black patches originating from the color of magnetite. The typical histopathological changes in the lungs of the treated rats included infiltration of macrophages phagocytosing magnetite, inflammatory cell infiltration, granuloma formation and an increase of goblet cells in the bronchial epithelium. The results clearly show that instilled magnetite causes foreign body inflammatory and granulating lesions in the lung. These pulmonary responses occur in a dose-dependent manner in association with the increase in lung weight.     

Efficacy and safety of lomefloxacin on bacterial extraocular disease in the horse

Lomefloxacin is a broad-spectrum fluoroquinolone antibiotic used for the treatment of bacterial extraocular disease. This study investigated the efficacy and safety of lomefloxacin eye drops for bacterial extraocular disease in horses. Lomefloxacin ophthalmic solution (0.3%) was instilled three times daily for 2–5 days in 65 horses diagnosed with bacterial extraocular disease based on clinical findings. Clinical observations and bacteriological examinations were performed at the start of treatment, 2 and 5 days after the start of treatment, and at the discontinuation or termination of treatment. Of the 65 horses, 64 were positive for bacteria, and 22 bacterial genera and 47 bacterial species were identified. The efficacy of lomefloxacin was evaluated in 63 horses; one horse with a negative culture and another with suspected bacterial contamination were excluded. Lomefloxacin was considered to be clinically effective in 54 horses. The major bacterial species identified were Staphylococcus aureus, Streptococcus equi subsp. zooepidemicus, Acinetobacter lwoffii, Staphylococcus xylosus, Staphylococcus vitulinus, Enterobacter agglomerans, Flavimonas oryzihabitans and Staphylococcus sciuri, with a cumulative disappearance rate of 80% or more at the termination of instillation. Excluding one horse that did not undergo a bacteriological examination, the remaining 62 horses were assessed for bacteriological outcome. Full or partial bacterial clearance was detected in 95% or more of the 62 horses. One of the 65 horses reported adverse events that had no causal relation with the eye drops. Our results showed that lomefloxacin is safe and effective for the treatment of bacterial extraocular disease in horses.     

Azaphilones, furanoisophthalides, and amino acids from the extracts of Monascus pilosus-fermented rice (red-mold rice) and their chemopreventive effects

Six azaphilones, monascin (1), ankaflavin (2), rubropunctatin (3), monascorburin (4), rubropunctamine (5), and monascorburamine (6), two furanoisophthalides, xanthomonasin A (7) and xanthomonasin B (8), and two amino acids, (+)-monascumic acid (9) and (-)-monascumic acid (10), isolated from the extracts of Monascus pilosus-fermented rice (red-mold rice) were evaluated for their inhibitory effects on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in mice, on the induction of Epstein-Barr virus early antigen (EBV-EA) by TPA in Raji cells, and on the activation of (+/-)-(E)-methyl-2[(E)-hydroxy-imino]-5-nitro-6-methoxy-3-hexemide (NOR 1), a nitric oxide (NO) donor. Among the compounds tested, seven compounds (1-6 and 10) on TPA-induced inflammation, and six compounds (1, 3-5, 9, and 10) on EBV-EA activation, exhibited potent inhibitory effects. All of the compounds tested showed moderate inhibitory effects on NOR 1 activation.     

Responses of fees used for revegetation of desert areas to iron deficiency stress

We investigated the responses to iron deficiency of four major tree species, Prosopis cineraria (local name: Ghaf), Acacia tortilis (Samar), Zizyphus spina-christi (Sidr), and Leptadenia pyrotechnica (Markh), used for revegetation of desert areas in the United Arab Emirates (UAE). The responses to iron deficiency differed among the tree species. Ghaf, Samar, and Sidr decreased the nutrient solution pH in response to iron deficiency. Markh and iron-deficient Sidr did not release protons in the CaCl₂ solution. The Fe reducing capacity of the roots of Ghaf, Samar, and Sidr was significantly enhanced by iron deficiency, whereas a smaller increase in the reducing activity of the roots of Markh was observed. In all the tree species, the amount of reductant released from iron-deficient plants was higher than that from iron-sufficient ones. Markh released a small amount of reductant under irondeficient conditions. In the present study, the reductant released by all the trees was caffeic acid. Acidification treatment enhanced the amount of reductant released irrespective of iron treatments in Ghaf, Samar, and Sidr but had no effect on the amount of reductant released in iron-deficient Markh. The activity of p-coumarate hydroxylase was higher in Sidr and Ghaf than in the other trees. This activity decreased in iron-deficient Samar. Ghaf, Samar, and Sidr showed a high capacity to lower the pH of the nutrient solution, and Ghaf markedly enhanced the effectiveness of all the examined mechanisms, while Markh showed a lower ability compared to the other tree species.     

Inhibitory effects of cucurbitane glycosides and other triterpenoids from the fruit of Momordica grosvenori on epstein-barr virus early antigen induced by tumor promoter 12-O-tetradecanoylphorbol-13-acetate

Two new triterpene benzoates, 5-dehydrokarounidiol dibenzoate (1) and karounidiol dibenzoate (2), and two new triterpene glycosides, 5alpha,6alpha-epoxymogroside IE(1) (8) and 11-oxomogroside A(1) (9), along with 15 known triterpenoids (one triterpene benzoate, 3; three triterpene mono-ols, 4-6; one triterpene aglycon, 7; and 10 triterpene glycosides, 10-19), were isolated from the ethanol extract of the fruit of Momordica grosvenori. The structures of 1, 2, 8, and 9 were determined on the basis of spectroscopic and chemical methods. Among the known triterpene glycosides, mogroside I E(1) (12) was a new naturally occurring compound. Eighteen triterpenoids (2-19) and 11-oxomogrol (20), a hydrolysis product of 9, were evaluated with respect to their inhibitory effects on the induction of Epstein-Barr virus early antigen (EBV-EA) by 12-O-tetradecanoylphorbol-13-acetate (TPA) in Raji cells, which is known to be a primary screening test for antitumor promoters. All of the compounds tested showed potent inhibitory effects on EBV-EA induction (70-100% inhibition at 1 x 10(3) mol ratio/TPA).     

Effects of helical angle of router bit on acoustic emission

The purpose of this study was to investigate the effects of changes in the helical angle of the router bit on the acoustic emission (AE) signal for various workpiece grain angles. The helical angle varied from 0° to 50° at 10° increments. The workpiece grains were oriented from 0 degrees (cutting parallel to the grain), through 90° (cutting end of the grain), to 165° at 15° increments. The AE signals and machined surface roughness were measured in an attempt to clarify the relations between them. The results were summarized as follows: (1) The AE signals were lowest in the with the grain cutting zones and slightly increased in the against the grain cutting zones; they rapidly reached the highest values at the 135°–165° grain angles. The greater the helical angle of the router bit, the smaller were the AE count rates for each grain angle investigated. There was no significant change in AE generation for helical angles of 0° and 10°. Moreover, the greater the feed rate, the greater was the AE count rate for every cutting condition investigated. (2) The surface roughness, similar to the AE count rate, had the lowest values in the with the grain cutting zones, slightly increased until the 120° grain angle, and then rapidly become extremely rough, reaching a maximum at the grain angles of 135°–150°. There was no remarkable change in the machined surface roughness while routing with the grain using the router bit of greater helical angle. However, when routing against the grain, the greater the helical angle the smoother was the machined surface. (3) There were correlations between the AE count rate and the machined surface roughness for each helical angle investigated. Therefore, acoustic emission has shown promise for monitoring and controlling the routing operation, including various grain angles and helical angles of the router bit.     

Transport and survival of Japanese sardine (Sardinops melanostictus) eggs and larvae via particle‐tracking experiments

Particle‐tracking experiments were performed to infer the distribution of larvae of the Japanese sardine (Sardinops melanostictus) and to detect effects of transport environment on sardine recruitment, using the output of a high‐resolution ocean general circulation model and observed data of sardine spawning grounds during 1978–2004. By the 60th day following spawning, approximately 50% of the larvae had been transported to the Kuroshio Extension (KE). Whereas the spawning period and grounds changed markedly in relation to the stock level, the proportion of larvae transported to the KE remained relatively constant and no significant correlations were found between sardine recruitment and the transport proportion. Instead, the recruitment was found to be correlated with physical parameters including the mixed layer depth and the sea surface temperature along several major transport trajectories of sardine larvae. The correlations were most significant for the trajectories in the region 0.5° south to 1° north of the Kuroshio axis (defined as the location of velocity maxima at each longitude) and for larvae spawned in February and March during the high stock period (1978–94), and for larvae spawned in March and April during the low stock period (1995–2004).     

Induction of apoptosis on K562 cell line and double strand breaks on colon cancer cell line expressing high affinity receptor for granulocyte macrophage-colony stimulating factor (GM-CSF)

BACKGROUND: Immunotoxins are comprised of both the cell targeting and the cell killing moieties. We previously established a new immunotoxin, i.e. Shiga toxin granulocyte macrophage-colony stimulating factor (StxA1-GM-CSF), comprises of catalytic domain of Stx, as a killing moiety and GM-CSF, as a cell targeting moiety. In this study, the ability of the immunotoxin to induce apoptosis and double strand breaks (DSB) on different cell lines was investigated. METHODS: The recombinant hybrid protein was expressed in bacterial expression system and purified with nickel-nitrilotriacetate acid resin. The K562 (erythroid leukemia) cell line and LS174 (colon carcinoma) were used in this study. The neutral comet assay was carried out for the detection of DSB and Hoechst staining was performed for apoptosis. RESULTS: StxA1-GM-CSF effectively induced apoptosis on K562 cell line and DNA Double Strand Break (DSB) were observed on colon cancer cell line treated with StxA1-GM-CSF. CONCLUSION: This novel action i.e. DNA damage might be a relevant mechanism of action for StxA1-GM-CSF that is designed to act as immunotoxin, although further investigation is required.     

LED不同光质对草莓组织培养的影响

以京藏香草莓(Fragaria ananassa Duch.cv.Jingzangxiang)匍匐茎茎尖为试验材料,以LED白光为对照,研究了红光、蓝光、组合光(红∶蓝∶远红=1∶1∶1)不同LED光质对草莓茎尖组织培养的影响。结果表明,在茎尖诱导分化阶段,丛生芽分化时间快慢依次为红光组合光白光蓝光;诱导分化率高低依次为组合光白光红光蓝光;增殖系数大小依次为组合光白光红光蓝光;生根率高低依次为组合光白光蓝光红光,植株生长势强弱依次为组合光白光红光蓝光。综合而言,LED组合光质有利于草莓茎尖组织培养。