Gas-liquid chromatographic determination of thiourea in citrus peels

A gas-liquid chromatographic (GLC) method was developed for the detection and determination of thiourea in citrus peels. After the peel is extracted with ethyl ether, the ether extract is adsorbed on sodium sulfate together with water. Thiourea is recoverd from both the sodium sulfate and the peel residue with ethyl acetate-acetone(2+1). The extracted mixture is cleaned on an alumina column, the eluate is concentrated under vacuum, and thiourea is extracted from the concentrate with sodium carbonate solution. GLC was carried out on the prepared benzoyl derivative of thiourea. The average recoveries of thiourea from lemon peel were 85.3, 93.1, and 97.6% at the fortification levels of 1, 10, and 100 ppm, respectively. The detection limit was low as 0.08 ppm.     

Early detection of urinary bladder carcinogens in rats by immunohistochemistry for γ-H2AX: a review from analyses of 100 chemicals

In safety evaluations of chemicals, there is an urgent need to develop short-term methods to replace long-term carcinogenicity tests. We have reported that immunohistochemistry for γ-H2AX, a well-established biomarker of DNA damage, can detect bladder carcinogens at an early stage using histopathological specimens from 28-day repeated-dose oral toxicity studies in rats. Given the markedly low level of γ-H2AX formation in the bladder urothelium of untreated rats, an increase in γ-H2AX-positive cells following chemical exposure can be relatively easy to identify. Among the 100 compounds examined to date, bladder carcinogens can be detected with high sensitivity (33/39; 84.6%) and specificity (58/61; 95.1%). As expected, γ-H2AX formation levels tended to be high following exposure to genotoxic bladder carcinogens, whereas nongenotoxic bladder carcinogens also increased the number of γ-H2AX-positive cells, probably through secondary DNA damage associated with sustained proliferative stimulation. γ-H2AX formation in the bladder urothelium reflects species differences in susceptibility to bladder carcinogenesis between rats and mice and shows a clear dose-dependency associated with the intensity of tumor development as well as high reproducibility. Some of the bladder carcinogens that showed false-negative results in the evaluation of γ-H2AX alone could be detected by combined evaluation with immunostaining for bladder stem cell markers, including aldehyde dehydrogenase 1A1. This method may be useful for the early detection of bladder carcinogens, as it can be performed by simple addition of conventional immunostaining using formalin-fixed paraffin-embedded tissues from 28-day repeated-dose toxicity studies in rodents, which are commonly used in safety evaluations of chemical substances.     

A hairy root culture of melon produces aroma compounds

Musk melon is the favorite fruit with a high market value in Japan, and the fragrance is one of the major factors determining the fruit quality of melon. In this study, mutant melon hairy roots which had been induced by means of the T-DNA insertion mutagenesis were found to produce volatile compounds with the fruity fragrance of mature melon. The volatile compounds were extracted and identified by GLC-mass spectrometry. Some essential oils such as (Z)-3-hexenol, (E)-2-hexenal, 1-nonanol, and (Z)-6-nonenol were stably synthesized by these hairy roots despite the increased number of subcultures. The productivity of these compounds by the best hairy root line was shown to be considerably higher than naturally ripened melon fruits.     

豆浆通电加热过程有限元解析与验证

在通电加热加工充填豆腐的过程中,豆浆内部的温度分布对豆腐质构均匀性有很大的影响.研究豆浆通电加热过程中内部温度的分布及变化,有利于豆腐加热凝固系统的研发.对通电加热槽内豆浆的连续通电加热过程进行有限元二维模拟,并进行了实验验证.结果表明,通电加热过程中豆浆的电导率与温度之间呈线性关系;升温速率随着温度的增加而增加;豆浆内部温度分布比较均匀,靠近加热槽壁的豆浆温度较低,且随着加热时间从50 s增加到300 s,豆浆的最大温差从3℃增大到20.2℃.模拟结果和实验结果之间的均方根误差为1.86％,利用有限元模拟很好地预测了通电加热过程中豆浆内部的温度分布和变化.     

Somatic cell count determination in cow's milk by near-infrared spectroscopy: a new diagnostic tool.

The potential of near-infrared spectroscopy (NIR) in the region from 1,100 to 2,500 nm to measure somatic cell count (SCC) content of cow's milk was investigated. A total of 196 milk samples from seven Holstein cows were collected for 28, consecutive days, starting from 7th d after calving, and analyzed for fat, protein, lactose, and SCC. Three of the cows were healthy, and the remainder had periods of mastitis during the experiment. Near-infrared transflectance milk spectra were obtained using an InfraAlyzer 500 spectrophotometer. The calibration for logSCC was performed using partial least square (PLS) regression and different spectral data pretreatment. The best accuracy of determination was found for an equation that was obtained using smoothed absorbance data and 10 PLS factors. The standard error of calibration was 0.361, the calibration coefficient of multiple correlation was 0.868, the standard error of prediction for independent validation set of samples was 0.382, the correlation coefficient was 0.854, and the coefficient of variation was 7.63%. The accuracy of logSCC determination by NIR spectroscopy would allow health screening of cows and differentiation between healthy and mastitic milk samples. It has been found that SCC determination by NIR milk spectra is based on the related changes in milk composition. The most significant factors that simultaneously influenced milk spectra with the elevation of SCC were alteration of milk proteins and changes in ionic concentration of milk.     

Domestication and cereal feeding developed domestic pig‐type intestinal microbiota in animals of suidae

Intestinal microbiota are characterized by host‐specific microorganisms, which have been selected through host‐microbe interactions under phylogenetic evolution and transition of feeding behavior by the host. Although many studies have focused on disease‐related intestinal microbiota, the origin and evolution of host‐specific intestinal microbiota have not been well elucidated. Pig is the ideal mammal model to reveal the origin and evolution of host‐specific intestinal microbiota because their direct wild ancestor and close phylogenetic neighbors are available for comparison. The pig has been recognized as a Lactobacillus‐type animal. We analyzed the intestinal microbiota of various animals in Suidae: domestic pigs, wild boars and Red river hogs to survey the origin and evolution of Lactobacillus‐dominated intestinal microbiota by metagenomic approach and following quantitative PCR confirmation. The metagenomic datasets were separated in two clusters; the wild animal cluster being characterized by a high abundance of Bifidobacterium, whereas the domesticated (or captured) animal cluster by Lactobacillus. In addition, Enterobacteriaceae were harbored as the major family only in domestic Sus scrofa. We conclude that domestication may have induced a larger Enterobacteriaceae population in pigs, and the introduction of modern feeding system further caused the development of Lactobacillus‐dominated intestinal microbiota, with genetic and geographical factors possibly having a minor impact.     

Infectivity of feline enteroepithelial stages of Toxoplasma gondii isolated by Percoll-density gradient centrifugation.

The infectivity of the feline enteroepithelial stages of Toxoplasma gondii isolated by Percoll-density gradient centrifugation was examined by the trypan blue dye exclusion method by assaying their penetration into feline fibroblast cells in vitro and by inoculation of the intestinal mucosa of cats. A large population of the parasites showed trypan blue dye exclusion activity. When feline fibroblast cells were inoculated with feline enteroepithelial stage parasites, no intracellular parasites were found 18 h post-inoculation. Kittens inoculated intraduodenally with 2 x 10(6) feline enteroepithelial stage parasites shed oocysts between 2 and 8 days post-inoculation. These results indicate that the isolated feline enteroepithelial stage parasites display infectivity towards enterocytes of cats and are capable of gametogenesis.     

Localization and responsiveness of a cowpea apyrase VsNTPase1 to phytopathogenic microorganisms

Apyrases (NTPases) are associated with both compatible and incompatible interactions between plants and microorganisms. Previously we reported that the ATPase activities of cell-wall-bound apyrases of several leguminous plants, such as pea, cowpea, soybean, and kidney bean, were enhanced by a glycoprotein elicitor and were inhibited in a species-specific manner by mucin-type glycopeptide suppressors secreted from a pea pathogenic fungus, Mycosphaerella pinodes. In this study, we isolated two apyrase genes, VsNTPase1 and VsNTPase2, from a cDNA library of Vigna sinensis Endl. cv. Sanjakusasage. Based on phylogenetic analysis, VsNTPase1 may belong to a group that responds to environmental stimuli. In a transient assay using DNA bombardment, a fusion protein of green fluorescent protein (GFP) and the N-terminal putative signal sequence of VsNTPase1 was distributed in the nucleus, cytoplasm (cytoskeletal structure), and cell wall. On the other hand, a fusion protein of GFP and the N-terminal putative VsNTPase2-signal sequence was localized in the cytoplasm, especially in small particles (perhaps mitochondria). A recombinant VsNTPase1 expressed in Spodoptera frugiperda 21 cells responded directly to signal molecules from several phytopathogenic microorganisms. Here, we discuss the role of apyrases in recognizing and responding to exogenous signals. The nucleotide sequences of VsNTPase1 and VsNTPase2 in this article have been submitted to DDBJ as accession numbers AB196769 and AB196770, respectively.