TGF-β and TNF-α stimulate MMP-9 production in murine epidermal keratinocytes

Matrix metalloproteinases 2 and 9 (MMP‐2 and ‐9) are zinc‐dependent metalloenzymes and have gelatin‐degrading activity. Both MMP are known to be secreted by many types of cells and play important roles in several biological changes including tissue remodeling and wound healing. In the present study, a primary culture of murine epidermal keratinocytes was prepared and effects of transforming growth factor‐β (TGF‐β), tumor necrosis factor‐α (TNF‐α) and interferon‐γ (IFN‐γ) on expression of MMP‐2 and MMP‐9 by the keratinocytes was examined. Gelatin zymography revealed that murine epidermal keratinocytes secreted proenzyme forms of MMP‐2 and MMP‐9, but the active forms of both MMP were hardly detectable, indicating that in vitro autoactivation of these proenzymes did not occur. Both TGF‐β and TNF‐α stimulated MMP‐9 production in a dose‐dependent manner, but the MMP‐2 level was not changed. Interferon‐γ hardly affected production of MMP‐2 or MMP‐9. Ribonuclease protection assay demonstrated that TNF‐α increased the level of MMP‐9 mRNA 6‐fold compared to the control, whereas TGF‐β slightly up‐regulated it. These results suggest that expression of MMP‐9 could be regulated by several cytokines in murine epidermal keratinocytes.     

Vertical profiles of environmental factors within tropical rainforest, Lambir Hills National Park, Sarawak, Malaysia

Environmental factors, such as global solar radiation, wind speed, air temperature, humidity, and CO₂ concentration, were measured above and within the canopy of a tropical rainforest in Lambir Hills National Park, Sarawak, Malaysia. Few data concerning the environment of this forest have been reported. Intensive observations were carried out in 1998, 1999, and 2000 with the following results: (1) The fraction of global solar radiation reaching the upper layer of the canopy varied with global solar radiation above the canopy. Even though the global solar radiation above the canopy fluctuated, the fraction of that reaching the lower canopy and the ground was constantly approximately 5%. (2) The fraction of wind speed reaching each layer of the canopy increased with wind speed above the canopy. Little wind was usually present at the lower canopy. (3) The daytime air temperature at the canopy top was higher than that near the ground. The maximum difference between the air temperature at the canopy top and that at the ground was about 5°C, and the diurnal temperature ranges at the canopy top and those at the ground were about 8°C and about 5°C, respectively. The highest daytime water vapor pressure occurred within the canopy and particularly near the ground. Vertical gradients of water vapor pressure during the day were steep, probably because of high transpiration. (4) In the 1998 observation the minimum and the maximum CO₂ concentrations were 360 ppm in the day and 450 ppm at night, while in the 2000 observation the minimum and the maximum CO₂ concentrations were 350 ppm in the day and 540 ppm at night. The higher CO₂ concentration in the daytime and the lower concentration at night observed during the 1998 observation period were probably due to reduced photosynthesis and soil respiration caused by exceptional dry conditions during the observation period.     

Genetic variants of the unsaturated fatty acid receptor GPR120 relating to obesity in dogs

G protein-coupled receptor (GPR) 120 is an unsaturated fatty acid receptor, which is associated with various physiological functions. It is reported that the genetic variant of GPR120, p.Arg270His, is detected more in obese people, and this genetic variation functionally relates to obesity in humans. Obesity is a common nutritional disorder also in dogs, but the genetic factors have not ever been identified in dogs. In this study, we investigated the molecular structure of canine GPR120 and searched for candidate genetic variants which may relate to obesity in dogs. Canine GPR120 was highly homologous to those of other species, and seven transmembrane domains and two N-glycosylation sites were conserved. GPR120 mRNA was expressed in lung, jejunum, ileum, colon, hypothalamus, hippocampus, spinal cord, bone marrow, dermis and white adipose tissues in dogs, as those in mice and humans. Genetic variants of GPR120 were explored in client-owned 141 dogs, resulting in that 5 synonymous and 4 non-synonymous variants were found. The variant c.595C>A (p.Pro199Thr) was found in 40 dogs, and the gene frequency was significantly higher in dogs with higher body condition scores, i.e. 0.320 in BCS4–5 dogs, 0.175 in BCS3 dogs and 0.000 in BCS2 dogs. We conclude that c.595C>A (p.Pro199Thr) is a candidate variant relating to obesity, which may be helpful for nutritional management of dogs.     

Expression of matrix metalloproteinases in bovine luteal cells induced by prostaglandin F2α, interferon γ and tumor necrosis factor α

We recently demonstrated that luteal cells flow out from the ovary via lymphatic vessels during luteolysis. However, the regulatory mechanisms of the outflow of luteal cells are not known. Matrix metalloproteinases (MMPs) can degrade the extracellular matrix and basal membrane, and tissue inhibitors of matrix metalloproteinases (TIMPs) inhibit the activity of MMPs. To test the hypothesis that MMP expression in luteal cells is regulated by luteolytic factors, we investigated the effects of prostaglandin F2α (PGF), interferon γ (IFNG) and tumor necrosis factor α (TNF) on the mRNA expression of MMPs and TIMPs in cultured luteal cells. Luteal cells obtained from the CL at the mid-luteal stage (days 8–12 after ovulation) were cultured with PGF (0.01, 0.1, 1 μM), IFNG (0.05, 0.5, 5 nM) and TNF (0.05, 0.5, 0.5 nM) alone or in combination for 24 h. PGF and IFNG significantly increased the expression of MMP-1 mRNA. In addition, 1 μM PGF in combination with 5 nM IFNG stimulated MMP-1 and MMP-9 mRNA expression significantly more than either treatment alone. In contrast, IFNG significantly decreased the level of MMP-14 mRNA. The mRNA expression of TIMP-1, which preferentially inhibits MMP-1, was suppressed by 5 nM INFG. One μM PGF and 5 nM IFNG suppressed TIMP-2 mRNA expression. These results suggest a new role of MMPs: luteal MMPs stimulated by PGF and IFNG break down the extracellular matrix surrounding luteal cells, which accelerates detachment from the CL during luteolysis, providing an essential prerequisite for outflow of luteal cells from the CL to lymphatic vessels.     

Lipid distribution in branching coral Montipora digitata

>ABSTRACT: The lipid profile was studied along the branch length, from the top, middle to base portion, of coral Montipora digitata to gain more insight into the physiological significance of lipids in the coral energy budget. The lipids of M. digitata consisted of seven major lipid classes: polar lipid, sterol, free fatty acid, unknown lipids 1 and 2, triacylglycerol (TG), and wax ester. The concentration of storage lipids, TG, and wax ester showed a top–base gradient along the length, whereas the levels of free fatty acid and unknown lipids showed a base–top gradient. The proportions of polar lipid and sterol in the top portion of the branch were slightly higher than those in the base portion. This observation appeared to be compatible with the view that the increased energy expenditure for proliferation enhanced the mobilization of the storage fuel lipids of wax ester and TG rather than the structure lipids of polar lipids and sterols at the top portion of the branch. Compositions of fatty acid also showed a length-wise diversity. The top portion had a lower proportion of palmitic acid (16   :   0) in all lipid classes of fatty acid ester, suggesting that this fatty acid was preferentially mobilized at the top portion, probably for the growth of coral cells.     

Effect of Application of Acidified Porous Hydrate Calcium Silicate and Porous Hydrated Calcium Silicate on the Growth of Rice Plants (Oryza sativa L.)

The effect of the application of acidified porous hydrate calcium silicate (APS) in nursery bed soil and porous hydrate calcium silicate (PS) in paddy fields on the growth of rice plants ( Oryza sativa L. cv. Hitomebore) was examined in 2002 and 2003. The results revealed the following: 1) Shoot dry weight of rice seedlings increased by APS treatment in nursery bed soil. The tiller number of rice plants after transplanting in both years also increased by APS treatment in nursery bed soil, and in 2003, the tiller number in the treatment with a combination of APS in nursery bed soil and PS in paddy fields was significantly higher than that in the other treatments until the maximum tiller number stage. Furthermore, the root length of rice plants 14 d after transplanting increased by APS treatment in nursery bed soil. 2) Silicon concentration in the soil solution significantly increased by PS treatment in paddy fields, and the concentration of dissolved carbon oxide increased by APS treatment in nursery bed soil. 3) Only in the APS treatment the rice yield was 341 g m⁻², while 400 and 450 g m⁻² in the PS and both APS and PS treatments, respectively, in 2003. Percentages of ripened grains in the plots without PS treatment ranged from 57 to 63%, respectively, while, those in the PS treated plots were 82%. The numbers of panicles and ripened grains in both APS and PS treatments were the highest among the treatments. Based on the above results, we concluded that both APS in nursery bed soil and PS in paddy field treatments were effective in improving the silicon nutrition and growth of rice plants, and that this effect was enhanced by a combination of treatments with the two.     

Development of wood and wood ash-based hydroxyapatite composites and their fire-retarding properties

For making efficient use of waste wood ash emitted from wood biomass plant, the wood and wood ash-based hydroxyapatite (HAp) composite was produced and their flammability characterization was studied by thermogravimetric (DTA-TG) analysis, oxygen index (OI) measurement and cone calorimeter test. The results show that the exothermic and weight loss peaks in DTA-TG combustion profiles due to their significant thermal decomposition were weakened by the HAp agent impregnation. In addition, the OI value of HAp composites was increased by the HAp combining and the OI showed a correlation with the HAp contents. Also, the cone calorimeter study revealed that the heat release rates were decreased with increasing amount of HAp injection and accordingly their total heat release has an inverse relationship to the HAp contents. These results indicate that the treatment with wood ash-based HAp agents can enhance the flame retardancy of the treated woods.     

Determination of antioxidant activity and characterization of antioxidant phenolics in the plum vinegar extract of cherry blossom (Prunus lannesiana)

Sakura-cha (salted cherry blossom tea) is a Japanese tea that is traditionally served at celebrations such as wedding ceremonies. The production of Sakura-cha includes the immersion of cherry blossom flowers in Japanese plum vinegar, and through this process, the byproduct (plum vinegar extract of cherry blossom) is obtained. In this study, the antioxidant activity of the plum vinegar extract of cherry blossom was examined. The plum vinegar extract of cherry blossom had a greater superoxide anion scavenging activity compared with red wine, which is a well-known strong antioxidant. Liquid chromatography-mass spectrometry analysis showed that cyanidin-3-glucoside, cyanidin-3-rutinoside, and caffeic acid were the major components in the phenolic extract prepared from plum vinegar extract of cherry blossom, and they possessed superoxide anion scavenging activity. Caffeic acid is mainly responsible for the scavenging activity of phenolic extract; the contributions of cyanidin-3-glucoside and cyanidin-3-rutinoside were minor.     

Advanced maternal age induces fetal growth restriction through decreased placental inflammatory cytokine expression and immune cell accumulation in mice

Advanced maternal age is a risk factor for female infertility, and placental dysfunction is considered one of the causes of pregnancy complications. We investigated the effects of advanced maternal aging on pregnancy outcomes and placental senescence. Female pregnant mice were separated into three groups: young (3 months old), middle (8–9 months old), and aged (11–13 months old). Although the body weights of young and middle dams gradually increased during pregnancy, the body weight of aged dams only increased slightly. The placental weight and resorption rate were significantly higher, and live fetal weights were reduced in a maternal age-dependent manner. Although mRNA expression of senescence regulatory factors (p16 and p21) increased in the spleen of aged dams, mRNA expression of p16 did not change and that of p21 was reduced in the placenta of aged dams. Using a cytokine array of proteins extracted from placental tissues, the expression of various types of senescence-associated secretory phenotype (SASP) factors was decreased in aged dams compared with young and middle dams. The aged maternal placenta showed reduced immune cell accumulation compared with the young placenta. Our present results suggest that models using pregnant mice older than 8 months are more suitable for verifying older human pregnancies. These findings suggest that general cellular senescence programs may not be included in the placenta and that placental functions, including SASP production and immune cell accumulation, gradually decrease in a maternal age-dependent manner, resulting in a higher rate of pregnancy complications.