Quantitative detection system for maize sample containing combined-trait genetically modified maize

Various countries have established regulations that stipulate the labeling of agricultural commodities, feed, and food products that contain or are made from genetically modified (GM) material or that contain adventitious GM material in amounts that exceed certain threshold levels. While regulations in some countries refer to GM material on a weight per weight (w/w) percentage, the currently applied detection methods do not directly measure the w/w percentage of the GM material. Depending on the particular method and the sample matrix it is applied to, the conversion of analytical results to a w/w percentage is challenging or not possible. The first rapid PCR system for GM maize detection on a single kernel basis has been developed. The equipment for the grinding of individual kernels and a silica membrane-based 96-well DNA extraction kit were both significantly revised and optimized for this particular purpose, respectively. We developed a multiplex real-time PCR method for the rapid quantification of GM DNA sequences in the obtained DNA solutions. In addition, a multiplex qualitative PCR detection method allows for the simultaneous detection of different GM maize traits in each kernel and thereby for identification of individual kernels that contain a combination of two or more GM traits. Especially for grain samples that potentially contain combined-trait GM maize kernels, the proposed methods can deliver informative results in a rapid, precise, and reliable manner.     

Surface Hardening of Carbon Steel Using High Powered YAG Laser

A high powered YAG laser with kaleidoscope for surface modification was applied to the surface hardening of carbon steels containing 0. 18-0.54 wt% C without the absorbents, and the relationships between laser processing and surface hardening were investigated by hardness and microstructure. The structure of the hardened zone underwent complete martensitic transformation in all of the carbon steels tested, and its hardness increased with greater carbon content. Under identical irradiated conditions, the hardened zone expanded with increasing carbon content. A hardened zone extending from the surface to a depth of 1.0 mm was obtained at a laser power of 1.0 kW and a scanning speed of 1 mm/sec. It was found that in the surface hardening of carbon steels, a high powered YAG laser can be used to control the hardened zone by selecting the appropriate irradiation conditions, however, the hardened zone was affected by the assistant gas and the flow rate.     

Paxillin isoforms in mouse. Lack of the gamma isoform and developmentally specific beta isoform expression

Paxillin, a focal adhesion protein, exists as multiple isoforms in humans (alpha, beta, and gamma). To understand more about the physiological role of each isoform, we have employed the mouse system. We found that although the alpha and beta isoforms are present in the mouse, the gamma isoform is not. The alpha isoform protein was detected clearly in most adult tissues, whereas the beta isoform protein was almost undetectable except in spleen, testis, thymus, and lung. On the other hand, mRNAs of both isoforms were detectable in all tissues we examined. High levels of the beta isoform protein was detected in peritoneal exudate macrophage cells in adult mouse as well as in cultured fibroblasts, together with the alpha isoform. The alpha isoform was expressed at a constant level throughout the embryonic stages we examined, whereas the beta isoform protein was detected at the mid-stages of development and increased to levels almost equal to those of the alpha isoform during the late stages of embryogenesis. Therefore, unlike the alpha isoform, expression of the beta isoform protein is restricted in adult tissues. Moreover, we showed that alpha and beta isoforms were colocalized within the same focal adhesion plaques, and cytoplasmic pools of both isoforms exist in the perinuclear area, colocalized with the Golgi apparatus.     

Silent myocardial ischemia in Kawasaki disease: evaluation of percutaneous transluminal coronary angioplasty by dobutamine stress testing

BACKGROUND: Myocardial ischemia and myocardial infarction are the most serious complications of coronary artery lesions in children with Kawasaki disease (KD). Therefore, early detection and treatment of myocardial ischemia in patients with KD is essential. We studied the effectiveness of percutaneous transluminal coronary angioplasty (PTCA) in patients with silent myocardial ischemia detected by dobutamine stress 99mTc myocardial scintigraphy (TMS), body surface mapping (BMS), and signal-averaged ECG late potentials (ELP). METHODS AND RESULTS: Eight of 76 asymptomatic patients with a coronary stenosis >25% and a positive dobutamine stress test were considered to have silent myocardial ischemia. All eight patients had >95% stenoses demonstrated by coronary angiography (CAG) just before PTCA. After PTCA, CAG showed that all of the coronary artery stenoses had been reduced to <50%. Additionally, intravascular ultrasonography (IVUS) performed in five patients before and after PTCA demonstrated adequate dilation of the coronary stenosis after PTCA. All eight patients underwent dobutamine stress TMS, BMS, and ELP 2 to 3 months after PTCA, which demonstrated no regions of myocardial ischemia. Approximately 6 months later, CAG was performed in all eight patients, and only one patient had developed restenosis. CONCLUSIONS: PTCA effectively dilates stenotic coronary arteries in children with KD. Moreover, dobutamine stress TMS, BMS, and ELP are useful for detecting silent myocardial ischemia and estimating the effectiveness of PTCA. Furthermore, IVUS is useful for evaluating the severity of coronary artery lesions before and after PTCA in patients with KD.     

Generation of metastatic variants of Eker renal carcinoma cell lines for experimental investigation of renal cancer metastasis

We and others have demonstrated that a mutation in Tsc2 is the rate-limiting step for renal carcinogenesis in the Eker rat model. Although inactivation of Tsc2 results in development of renal tumors, it is not sufficient for metastatic renal cell carcinomas (RCs) in the Eker rat. To investigate the additional genetic event(s) necessary for cancer metastasis, we have established highly metastatic S-Lk9d-SLM cell lines from a non-metastatic RC cell line (Lk9dL) by co-implantation with a foreign body (gelatin sponge). Since these cell lines were remarkably different in metastatic performance (all and none, respectively) despite having the same genetic background, they should be useful experimental tools to investigate metastasis-promoting events in renal carcinogenesis.     

Requirement for activation of the serine-threonine kinase Akt (protein kinase B) in insulin stimulation of protein synthesis but not of glucose transport

A wide variety of biological activities including the major metabolic actions of insulin is regulated by phosphatidylinositol (PI) 3-kinase. However, the downstream effectors of the various signaling pathways that emanate from PI 3-kinase remain unclear. Akt (protein kinase B), a serine-threonine kinase with a pleckstrin homology domain, is thought to be one such downstream effector. A mutant Akt (Akt-AA) in which the phosphorylation sites (Thr308 and Ser473) targeted by growth factors are replaced by alanine has now been shown to lack protein kinase activity and, when overexpressed in CHO cells or 3T3-L1 adipocytes with the use of an adenovirus vector, to inhibit insulin-induced activation of endogenous Akt. Akt-AA thus acts in a dominant negative manner in intact cells. Insulin-stimulated protein synthesis, which is sensitive to wortmannin, a pharmacological inhibitor of PI 3-kinase, was abolished by overexpression of Akt-AA without an effect on amino acid transport into the cells, suggesting that Akt is required for insulin-stimulated protein synthesis. Insulin activation of p70 S6 kinase was inhibited by approximately 75% in CHO cells and approximately 30% in 3T3-L1 adipocytes, whereas insulin-induced activation of endogenous Akt was inhibited by 80 to 95%, by expression of Akt-AA. Thus, Akt activity appears to be required, at least in part, for insulin stimulation of p70 S6 kinase. However, insulin-stimulated glucose uptake in both CHO cells and 3T3-L1 adipocytes was not affected by overexpression of Akt-AA, suggesting that Akt is not required for this effect of insulin. These data indicate that Akt acts as a downstream effector in some, but not all, of the signaling pathways downstream of PI 3-kinase.     

Surface wettability of silicon rubber after irradiation with a glow discharge plasma

Silicon rubber, PDMS, was irradiated by Ar, Ar/H₂ and Ar/O₂ plasmas to increase the surface wettability to water which was found to increase with the irradiation time for every application of plasma irradiation. The use of Ar/O₂ plasma was found to be particularly effective in enhancing the surface wettability. The aging behavior due to exposure to air, ethanol or water was also measured. In the case of continuous exposure to water after plasma treatment, the wettability could be maintained for more than several months with the water immersion.     

Formation process of Na-X zeolites from coal fly ash

In order to synthesize Na-X zeolite from coal fly ash (Fa), Fa was pretreated under stirring condition at various temperatures of 20–50°C for 72 h and then aged at 85°C for a given period with NaOH solutions. The resulting materials were characterized by various means. When Fa was aged for 72 h without pretreatment, species P were formed. As the pretreating temperature raised from 20 to 50°C, the degree of crystallinity of faujasite increased, while that of species P decreased. The faujasite species formed was identified as Na-X zeolite with molar ratio SiO₂/Al₂O₃ = 2.4. When Fa was pretreated at 50°C and aged for 60 h, the only species formed was Na-X zeolite. Increasing the pretreating temperature up to 50°C results in the increase of Si⁴⁺ and Al³⁺ concentrations in the treating solution by dissolution of amorphous material in Fa. With the conditions used, the crystalline phase, such as -quartz and mullite, was poorly dissolved during the treatment. Hence, the higher pretreating temperature would give the uniform nucleation and crystal growth of Na-X zeolite during the aging.     

Index for predicting post-operative residual liver function by pre-operative dynamic liver SPET

To predict residual liver function before hepatic resection, we devised a predictive index by combining clearance values with functional liver volume measured by liver dynamic single photon emission tomography (SPET). Forty-seven patients with liver disease underwent liver dynamic SPET with 99Tcm-Sn colloid before hepatic resection. There were no operation-related deaths. Three patients died from hepatic failure more than 1 month following the operation. Their predictive indices were 0.24, 0.33 and 0.34. When the predictive index was above 0.35, no patient had symptoms of hepatic failure or died. Our data suggest that when the predictive index is above 0.35, there is a low probability of hepatic failure after hepatectomy.     

Intragenic Tsc2 somatic mutations as Knudson''s second hit in spontaneous and chemically induced renal carcinomas in the Eker rat model

Membrane conductances during hypoosmotic swelling were characterized in rat astrocytes in primary tissue culture. Using whole cell patch clamp techniques, mean +/- SEM cell conductance in isoosmotic phosphate-buffered saline (PBS) was 55.6 +/- 5.8 pS/pF. Cell conductance (mean +/- SEM) increased from this initial value to 187 +/- 46%, 561 +/- 188%, and 1216 +/- 376% within 9 min of exposure to 220 mOsm, 190 mOsm, and 145 mOsm PBS, respectively. With each of these hypoosmotic exposures, no change occurred in membrane capacitance. When CsCl replaced KCl in the microelectrode solution, a similar conductance increase was obtained at each osmolality. However, when gluconate salts were used in place of chloride salts in the electrode solution, no significant conductance increase was observed with 190 mOsm PBS. With a KCl microelectrode solution, all conductance increase which occurred in 190 mOsm PBS was inhibited by 200 microM niflumic acid, but not by 5 mM BaCl(2). Both niflumic acid and BaCl(2) inhibited 60-80% of the conductance increase of cells in 145 mOsm PBS. Using a microelectrode solution containing taurine as the major anion, membrane conductance increased 5-fold when cells were placed in 250 mOsm medium. This conductance increase was completely inhibited by 200 microM niflumic acid. Thus, independent chloride and potassium conductances are activated by hypoosmotic swelling of cultured astrocytes while plasma membrane area is unaltered. The chloride conductance pathway is activated at a significantly lower degree of hypoosmotic exposure than that which activates the potassium pathway and may be permeable to anionic taurine. These conductance pathways may mediate diffusive loss of potassium, chloride, and taurine from these cells during volume regulation following hypoosmotic swelling.