The mTOR-regulated phosphoproteome reveals a mechanism of mTORC1-mediated inhibition of growth factor signaling

The mammalian target of rapamycin (mTOR) protein kinase is a master growth promoter that nucleates two complexes, mTORC1 and mTORC2. Despite the diverse processes controlled by mTOR, few substrates are known. We defined the mTOR-regulated phosphoproteome by quantitative mass spectrometry and characterized the primary sequence motif specificity of mTOR using positional scanning peptide libraries. We found that the phosphorylation response to insulin is largely mTOR dependent and that mTOR exhibits a unique preference for proline, hydrophobic, and aromatic residues at the +1 position. The adaptor protein Grb10 was identified as an mTORC1 substrate that mediates the inhibition of phosphoinositide 3-kinase typical of cells lacking tuberous sclerosis complex 2 (TSC2), a tumor suppressor and negative regulator of mTORC1. Our work clarifies how mTORC1 inhibits growth factor signaling and opens new areas of investigation in mTOR biology.     

Agrobacterium-mediated transformation of the CrDAT gene and selection of transgenic periwinkle lines with a high vincristine accumulation

Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg^?1 to 5.12 μg.mg^?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g^?1 (fresh weight) to 10.53 µg.g^?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.

Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter     

Volatile Flavor Components and the Body Weights of Rats in the Green Tea Added Miner Ingredients

In order to change for the good flavor in green tea of middle or low grade,roast naked barley （miner ingredient） was added to it.To make tea having functional property,mixed roast naked barley-green tea with one medicinal herb （MH,miner ingredient）.This study was performed to investigate the effect of drinking extracts of naked barley-MH-green tea （A group） and adlay-Angelica gigas Nakai-green tea （B group） on the body weights of rats,and volatile flavor components of best effective extract on weight reduction.Forty-eight male Sprague-Dawley rats divided into six groups were fed five different green tea extracts for 8 weeks.The result obtained in the experiment indicated that the change of body weights of rats was affected by A group,B group and others.A group showed highest reduction rate of body weights （36.40％）.Reduction rate of body weights on the B group （10.10％） was much lower than A group （36.40％）.Aroma compounds were extracted by the SDE method.The concentrated aroma extracts were analyzed and identified by GC and GC-MS.The main aroma components of the green tea blended with naked barley and MH were 12 pyrazines having roasty note,methyl butanals having sweet note,hexanal having briskness greenish note,and nerolidol and3-ionone having floral note.     

Effects of soil moisture content on CO2 triggered soil physicochemical properties in a near-surface environment

Journal of Soils and Sediments - Carbon capture and storage (CCS) has been frequently discussed as a strategy for meeting CO2 emission reduction and its targets. However, some critical issues have...     

Distribution and migration of Japanese common squid, Todarodes pacificus, in the southwestern part of the East (Japan) Sea

Data on squid catches of Korean and Japan, water temperatures at depth of 100 m, and night-visible images of fishing boats collected in the East (Japan) Sea from 1970 to 1999 were analyzed to examine the distribution and migration of the Japanese common squid, Todarodes pacificus, in the southwestern part of the East (Japan) Sea. The main fishing grounds detected from squid catch in each grid (0.5° latitude × 0.5° longitude) by Korean squid fishery and night-visible images provided by the Defense Meteorological Satellite Program (DMSP) Operational Linescan System (OLS) were situated in the southwestern part of the East (Japan) Sea. The distribution and migration route was illustrated from squid catch in each grid and DMSP OLS image. In years of high catches, the fishing grounds were situated mainly between Ulleung Island and the eastern coastal waters of Korea, while in years of low catches they were situated between Ulleung Island and the Yamato Bank in the central East (Japan) Sea. The center of fishing activity began moving northward from around the Korea/Tsushima strait to the northern boundary of the Tsushima Warm Current in March, reaching into the Yamato Bank in September, and then returning to the strait by February. The northward and southward migration routes differed; the northward migration route occurred closer to the mainland coast of Korea than the southward migration route did. This work suggests T. pacificus begin their northward migration almost 2 months earlier than previously suggested.     

Blood and liver biopsy for the non‐destructive screening of tilapia lake virus

Detection of tilapia lake virus (TiLV) in tilapines is mainly from visceral organs of killed fish. However, lethal sampling might not be viable to broodstock and economically important ornamental cichlids. To contribute towards screening of the virus in asymptomatic infected fish, a subclinically infected population of Nile tilapia adults obtained from a local farm was preliminarily tested to compare different non‐lethal sampling methods, for example liver biopsy, gill biopsy, fin clip, mucus, faeces and blood for detection of TiLV. Only liver and blood samples gave positive results by PCR. Since blood sampling is relatively simpler, it was further used for five naturally co‐cultured juvenile fish species from above‐mentioned farm including 40 red tilapia broodstock and 20 Nile tilapia adults from two other different farms. The results showed that from the tested fish, 4 of 5 Nile tilapia, 2 of 5 hybrid red tilapia and 3 of 5 giant gourami blood samples tested positive, while 38 of 40 blood samples of red tilapia tested positive for TiLV in second‐step PCR. Sequencing representative PCR amplicons of positive samples confirmed sequence identity to TiLV. In conclusion, both blood and liver biopsy are practical non‐destructive sampling platforms for TiLV screening in cichlids with blood being more convenient, especially for tilapia broodstock.     

Effect of dopamine on excretory urographic image quality and the prevention of contrast-induced nephropathy in dogs

The effect of low dose dopamine on the excretory urographic image quality and contrast media-induced nephropathy in normal dogs (experiment 1) and the dogs with decreased renal function (experiment 2) were assessed. In experiment 2, decreased renal function was induced by gentamicin overdose. In each experiment, animals were divided into 3 groups. In group 1, only contrast medium (iohexol) was administered. In group 2, contrast medium plus intravenous fluid (0.9% saline) were administered. And in group 3, contrast medium plus intravenous fluid and low dose dopamine were administered. Investigated parameters included intrarenal resistive index (RI), serum BUN and creatinine concentrations, contrast medium elimination time and radiographic image quality. In experiment 1, RI of group I increased at 80 min after contrast medium administration (p<0.05), but RI of group 3 decreased at 48 and 72 hr (p<0.05). Serum BUN concentration of group 1 was higher than that of group 2 and 3 (p<0.05); in radiographic examination, contrast medium elimination time decreased in group 2 and 3, but image quality of group 2 was inferior to that of group 3. In experiment 2, image quality of group 3 only provided adequate visualization of renal structures. The formula of contrast medium plus low dose dopamine was found to provide good nephrogram and pyelogram image quality without supplemental contrast medium, and to protect renal tubules from prolonged exposure to concentrated contrast medium.