Molecular screening of both the promoter and the protein coding regions in the human ob gene in Japanese obese subjects with non-insulin-dependent diabetes mellitus

OBJECTIVE: Although the molecular mechanism of obesity has been poorly understood, recent studies indicate that leptin plays a critical role in regulating both food intake and body weight. Because obesity decreases the sensitivity to insulin, the human ob gene is presumed to be one of the candidate genes for non-insulin-dependent diabetes mellitus (NIDDM) associated with obesity. Although the protein coding region in the ob gene has been screened for mutations, the promoter region and the non-coding first exon have not yet been studied. We investigated the involvement of the human ob gene, especially mutations at the promoter region and the non-coding first exon, in the development of NIDDM associated with obesity. SUBJECTS: The study group comprised 60 Japanese obese subjects with NIDDM (body mass index (BMI) 43.6 > or = BMI > or = 26.4, 29.0+/-0.41 (mean+/-S.E.M.)) and 24 obese individuals with impaired glucose tolerance (IGT) (30 > or = BMI > or = 26.4, 27.1+/-0.22). METHODS: Mutations at both the promoter region and all three exons in the human ob gene were screened by the single-stranded conformational polymorphism analysis. When aberrantly migrated bands were recognized, the PCR-amplified DNA fragment was directly sequenced. RESULTS: In the protein coding region a silent mutation in the second exon was detected. The non-coding first exon and the about 100 bp 5'-flanking region of the gene which contains a proximal CCAAT/enhancer-binding protein site were screened, but no mutations were found. CONCLUSION: These results suggest that no mutations in either the promoter region at the about 100 bp 5'-flanking region of the gene, or in any of the three exons, are involved in the development of NIDDM or IGT associated with obesity.     

Non-destructive determination of acid–brix ratio of tomato juice using near infrared spectroscopy

Consumer's acceptance of tomato juice depends on its sourness to sweetness ratio and measuring this is important for quality control and marketing. Traditional methods destroy the samples, are time consuming and cannot be used in continuous packing or bottling systems. A non‐destructive method of quality evaluation, using near infrared (NIR) techniques, was tested, by using a portable NIR measuring unit. Spectra of tomato juice of known acid and brix values were determined and, in the wavelength range 703–1124 nm (NIR), a calibration model for acid–brix ratio (ABR) was developed, by using unscrambler software. When used to predict ABR of tomato juice statistical analysis showed minimal standard error (0.009) and satisfactorily high correlation coefficients (0.92) over the wavelength range 1059.5–1124.8 nm, for both calibration and prediction. These values were hardly different from analytical results and the NIR model has potential for non‐destructive prediction of ABR of tomato juice.     

Effects of loading mode and water chemistry on stress corrosion crack growth behavior of 316L HAZ and weld metal materials in high temperature pure water

The stress corrosion cracking (SCC) growth rates of 316L weld heat-affected zone (HAZ) and weld metal materials in high temperature pure water at 288 °C were measured using contoured double cantilever beam (CDCB) specimens and an alternating current potential drop (ACPD) in situ crack-length monitoring system. The effects of loading mode and dissolved oxygen and hydrogen on crack growth rate (CGR) were experimentally quantified. Typical intergranular SCC was found in the HAZ specimen and interdendritic SCC was identified in the weld metal specimen. The HAZ specimen and the weld metal specimen showed quite a similar response to the applied loading modes and the water chemistry, even though their absolute CGR values were different. The crack growth rates under trapezoidal loading were moderately higher than those under constant loading by several tenths percent. Switching the water chemistry from the oxygen-bearing water to the hydrogen-bearing water drastically decreased the electrochemical potential and the crack growth rate, and vice versa. A time-lag period for crack growth was observed after switching the water chemistry back to the oxygen-bearing water, where the crack growth rate was low even the dissolved oxygen concentration and the electrochemical potential had become high. Strain hardening and the resultant uneven distribution of deformation contribute to the enhanced intergranular SCC growth behavior in the HAZ area. The crack growth kinetics is analyzed based on the deformation/oxidation interaction at the crack tip, considering the importance of the electric-charge transfer, mass transport kinetics and the crack tip strain rate.     

Interactive postprocessor and animation system using a microcomputer

This paper is concerned with the development of a microcomputer-aided graphic system. In this system the difficult task of computation for graphical representation is carried out on a large computer, whereas a microcomputer is used as the graphic terminal. Interface programs are newly developed for this purpose. A technique is proposed here to minimize the transfer time of graphic data from the large computer to the microcomputer. Finally, the animation and the interactive tomography systems are developed as extensions of the graphic system.     

Crystallization of nylon 6–clay hybrid by annealing under elevated pressure

Nylon 6–clay hybrid (NCH) is a molecular composite of nylon 6 and uniformly dispersed silicate layers of montmorillonite. We found that the phase with the high melting temperature (HMT phase) in the NCH annealed under elevated pressure. The melting temperature of the HMT phase was 240°C. Nylon 6 annealed under elevated pressure did not have the HMT phase. Thus, the presence of the HMT phase was characteristic of the NCH. The relative heat of fusion of the HMT phase (heat of fusion of HMT phase/heat of fusion in the pressure annealed NCH) increased with increase in pressure. High-pressure differential thermal analysis (DTA) measurement revealed that the temperature, at which the relative heat of fusion showed a maximum value, was below about 20°C of the melting temperature of the original NCH under elevated presssure. It was considered that the nylon 6 crystallite near the melting temperature and the molecular mobility under elevated pressure were necessary to the appearance to the HMT phase. © 1994 John Wiley & Sons, Inc.     

Influences of electroosmotic flows in nanopillar chips on DNA separation: Experimental results and numerical simulations

The various potential factors affecting the performance of nanopillar chips on DNA separation were investigated from the viewpoints of both numerical calculations and actual experiments. To yield higher performance and replace the conventional DNA separation techniques such as microchip electrophoresis, the phenomenon specific to the nanopillar chips should be deeply understood. In this paper, although various factors affecting the performance of the nanopillar chips are considered, we focused on the effect of electroosmotic flow, which is particularly noticeable in quartz-made nanopillar chips. High-resolution separation of DNA was realized when an electroosmotic flow was suppressed by simply using a higher concentration of buffer, but DNA separation failed in the presence of an electroosmotic flow. It was confirmed from the numerical simulations and the direct observations that the deformation of DNA band during the injection process was induced by electroosmotic flow and consequently led to a poor resolution.     

The dependency of the crack growth rate on the loading pattern and temperature in stress corrosion cracking of strain-hardened 316L stainless steels in a simulated BWR environment

The stress corrosion cracking (SCC) growth rate of a warm-rolled (WR) 316L stainless steel contoured double cantilever (CDCB) specimen was measured in high purity water at various temperatures and under various loading patterns. An alternating current potential drop (ACPD) technique was used to monitor the crack growth kinetics throughout the tests. The fracture surface exhibited typical intergranular SCC characteristics. Depending on the test conditions, three kinds of crack growth kinetics, i.e., increasing with time then becoming steady, being constant during the whole period, decreasing with time then becoming steady, were identified and are described. The steady state crack growth rate (CGR) values are used to quantify the effects of the loading pattern and the environmental temperature. A moderate increase in the crack growth rate was encountered by employing periods of unloading and reloading to form a trapezoidal loading pattern and the enhancement factor was found to depend on the holding time and the times for unloading and reloading. It was found that the crack growth is thermally activated; however, the apparent activation energy is not constant but seems to be greater at higher temperatures. Several types of temperature-dependent crack growth kinetics are proposed based on the rate-determining step for the crack growth. The present experimental results can be rationalized by considering multiple element processes such as aqueous mass transport and solid-state mass transport in the crack growth. The cracking mode, the temperature dependence of the crack growth rate, and the transient crack growth behavior for WR 316L SS after changing the environmental temperature are quite similar to those for a cold-worked(CW) 316L SS tested in the same environment, despite their different absolute crack growth rate values. The effect of yield strength on CGR is more significant at lower temperatures and the apparent activation energy for the crack growth rate seems to be lower in the material with a higher yield strength. Time-lag crack growth behavior was found at points during several test steps on WR 316L SS, for example, just after in situ pre-cracking and after increasing or decreasing the temperature, which is quite consistent with the results obtained with CW 316L SS. The importance of in situ monitoring of the crack growth for obtaining steady state crack growth rates is emphasized, especially for those steps for which a nonlinear crack growth period occurs after changing the test condition.