Changes in Chemical Structure and Biological Activity of L-DOPA as Influenced by an Andosol and Its Components

Velvetbean ( Mucuna pruriens ) has been reported to release 3-(3',4'-dihydroxyphenyl)-L-alanine (L-DOPA) as an allelochemical that inhibits the growth of other plants, although the inhibitory activity depends on the soil type and it is extremely reduced in Andosols. To clarify the effects of Andosols and their components on the chemical structure and plant-growth-inhibitory activity of L-DOPA, an L-DOPA solution was reacted with an Andosol and its components (weathered pumice and purified allophane), and the resultant solution was subjected to ¹H nuclear magnetic resonance and ultraviolet-visible spectral analyses, and plant-growth-inhibitory activity tests. When the L-DOPA solution was added to the soil components, the concentration of L-DOPA in the solution decreased by adsorption and transformation (polymerization) reactions. The adsorption mechanism included a ligand exchange reaction. The rate of L-DOPA transformation was faster at higher pH values. The soil components displayed a catalytic activity and accelerated the transformation of L-DOPA. Similar transformation occurred when light was irradiated. At pH values higher than 4.0, the transformed products from L-DOPA consisted of humic substances-like heterogeneous components, whereas specific components with low molecular weight were included when L-DOPA was transformed at a pH value of 9.7 or higher. The plant-growth-inhibitory activity of L-DOPA was extremely weakened when L-DOPA was adsorbed on or transformed (polymerized) by soil components. Therefore, in soils with high abilities of adsorption and transformation of L-DOPA such as in Andosols, it was likely that the L-DOPA concentration in the soil solution decreased quickly by adsorption and transformation reactions and the allelopathic activity of L-DOPA was lost.     

Development of taxon-specific sequences of common wheat for the detection of genetically modified wheat

Qualitative and quantitative Polymerase Chain Reaction (PCR) systems aimed at the specific detection and quantification of common wheat DNA are described. Many countries have issued regulations to label foods that include genetically modified organisms (GMOs). PCR technology is widely recognized as a reliable and useful technique for the qualitative and quantitative detection of GMOs. Detection methods are needed to amplify a target GM gene, and the amplified results should be compared with those of the corresponding taxon-specific reference gene to obtain reliable results. This paper describes the development of a specific DNA sequence in the waxy-D1 gene for common wheat (Triticum aestivum L.) and the design of a specific primer pair and TaqMan probe on the waxy-D1 gene for PCR analysis. The primers amplified a product (Wx012) of 102 bp. It is indicated that the Wx012 DNA sequence is specific to common wheat, showing homogeneity in qualitative PCR results and very similar quantification accuracy along 19 distantly related common wheat varieties. In Southern blot and real-time PCR analyses, this sequence showed either a single or a low number of copy genes. In addition, by qualitative and quantitative PCR using wx012 primers and a wx012-T probe, the limits of detection of the common wheat genome were found to be about 15 copies, and the reproducibility was reliable. In consequence, the PCR system using wx012 primers and wx012-T probe is considered to be suitable for use as a common wheat-specific taxon-specific reference gene in DNA analyses, including GMO tests.     

Detection of recombinant DNA segments introduced to genetically modified maize (Zea mays)

Polymerase Chain Reaction (PCR) techniques are increasingly used for the detection of genetically modified (GM) crops in foods. In this paper, recombinant DNAs introduced into the seven lines of GM maize, such as Event 176, Bt11, T25, MON810, GA21, DLL25, and MON802, are sequenced. On the basis of the obtained sequence, 14 primer pairs for the detection of the segments, such as promoter, terminator regions, and construct genes, were designed. To confirm the specificities of the designed primer pairs, PCR was performed on genomic DNAs extracted from GM and non-GM maize, GM and non-GM soy, and other cereal crops. Because the presence of the corresponding DNA segments was specifically detected in GM crops by the designed primer pairs, it was concluded that this method is useful for fast and easy screening of GM crops including unauthorized ones.     

Comparative studies of the quantification of genetically modified organisms in foods processed from maize and soy using trial producing

Seven types of processed foods, namely, cornstarch, cornmeal, corn puffs, corn chips, tofu, soy milk, and boiled beans, were trial produced from 1 and 5% (w/w) genetically modified (GM) mixed raw materials. In this report, insect resistant maize (MON810) and herbicide tolerant soy (Roundup Ready soy, 40-3-2) were used as representatives of GM maize and soy, respectively. Deoxyribonucleic acid (DNA) was extracted from the raw materials and the trial-produced processed food using two types of methods, i.e., the silica membrane method and the anion exchange method. The GM% values of these samples were quantified, and the significant differences between the raw materials and the trial-produced processed foods were statistically confirmed. There were some significant differences in the comparisons of all processed foods. However, our quantitative methods could be applied as a screening assay to tofu and soy milk because the differences in GM% between the trial-produced processed foods and their raw materials were lower than 13 and 23%, respectively. In addition, when quantitating with two primer pairs (SSIIb 3, 114 bp; SSIIb 4, 83 bp for maize and Le1n02, 118 bp; Le1n03, 89 bp for soy), which were targeted within the same taxon specific DNA sequence with different amplicon sizes, the ratios of the copy numbers of the two primer pairs (SSIIb 3/4 and Le1n02/03) decreased with time in a heat-treated processing model using an autoclave. In this report, we suggest that the degradation level of DNA in processed foods could be estimated from these ratios, and the probability of GM quantification could be experimentally predicted from the results of the trial producing.     

Study of the role of the carbohydrate and protein moieties of soy soluble polysaccharides in their emulsifying properties

The objective of this work was to investigate the role played by the protein fraction of soy soluble polysaccharide (SSPS) during its adsorption at oil/water interfaces. SSPS was separated in a high (HMF; 310 kDa) and low (LMF; 20 kDa) molecular weight fraction by gel filtration. SSPS/HMF consisted of 91.6% carbohydrate and 2.2% protein and showed better emulsifying properties than those of the whole SSPS, whereas SSPS/LMF seemed to affect negatively the adsorption behavior of SSPS. SDS-PAGE of the protein fraction obtained from SSPS/HMF showed a molecular mass of 50 kDa, was composed predominantly of proline (23.1%) and glutamic acid (15.2%), and still contained 8.8% of neutral sugar and 5.3% of uronic acid. Results indicated that not all of the protein material present in SSPS contributes to SSPS functionality but that only the material associated with HMF aids in the adsorption of SSPS onto oil/water interfaces.     

沙棘的化学成分和一氧化氮产物的抑制活性研究

基于对沙棘化学成份的调查，沙棘果中75％的乙醇提取物及其枝条中80％的丙酮氯仿可溶性物质可以抑制脂多糖（LPS）和重组小鼠的干扰素（IFN）-γ活性吞噬细胞即RAW264．7细胞中一氧化氮（NO）的生成。从果实提取物中可以分离出三种已知的黄酮、槲皮素（1）、kaempferol（2）、异鼠李素（3）和两种已知的萜类，果酸（4）和熊果酸（5）。该试验第一次从沙棘中提取出果酸，此外还从沙棘树皮的萃取物中分离出一种新的萜类、2-O-caffeoyl-谷酸（6），已知的萜类，鱼肝油醇酸（7），6-甲氧基-2H-1-苯芘（8）和β-醇（9）。并对RAW264．7细胞中的NO产物的活性抑制作用进行了研究。     

Histological evidence of lunar-synchronized ovarian development and spawning in the spiny rabbitfish Siganus spinus (Linnaeus) around the Ryukyus

ABSTRACT: This study clarifies the annual reproductive cycle and the lunar-synchronized spawning of the spiny rabbitfish ( Siganus spinus ) that inhabit the Okinawan waters. Annual and weekly changes in the gonadosomatic index (GSI) and the histological features of the ovaries were checked. Gonadosomatic index was high during the months of May to July, and yolk-laden oocytes were observed in the ovaries from March to July. Some of the ovaries collected during June and July contained oocytes at maturation stage or ovulatory follicles. These results suggest that the spiny rabbitfish undergo active vitellogenesis and spawning from May to July. During the reproductive season (May to July), collection of fish according to the lunar phase revealed that a high GSI occurred around the time of the new moon. Cyclic oocyte development with peaks around the time of the same moon phase was also observed, suggesting that, in Okinawan waters, this species is a lunar-synchronized spawner and spawns three times.     

Localization of a novel RNA-binding protein, SKIV2L2, to the nucleus in the round spermatids of mice

In our previous study (Kawashima et al., Biol Reprod 2009; 80: 1293-1304), we suggested that the first cycle of spermatogenesis recovered from busulfan-induced temporary arrest was a good model system to analyze the proteins expressed at the specific stages of spermatogenesis in the mouse, and this has been confirmed in the present paper. Namely, six-week-old mice were injected with busulfan at 20 mg/kg body weight. The germ cells except for the undifferentiated spermatogonia disappeared by 32 days after injection. The surviving spermatogonia started to proliferate, and spermatogenesis was entirely recovered about 77 days after injection. By proteome analysis of the busulfan-treated testis during the process of recovery of spermatogenesis, we identified a protein that was expected to be expressed in the spermatogenic cells as Superkiller viralicidic activity-2-like-2 (SKIV2L2). Skiv2l2 mRNA was found in the kidney, epididymis and heart as well as the testis. In the testis, Skiv2l2 mRNA was shown to be highly expressed in the spermatocytes at stages I to VI. On the other hand, SKIV2L2 protein was found to be predominantly localized in the nuclei of round spermatids. In accordance with the fact that SKIV2L2 belongs to the Ski2 family within the superfamily 2 of RNA helicases, it has been shown that SKIV2L2 has both the RNA-binding and ATPase activities.     

Ganglioneuroma in the urinary bladder of a dog

An 11-year-old male Labrador retriever presented with chronic oliguria. Ultrasonography findings revealed a protruding mass at the neck of the urinary bladder. A cystotomy was performed, and the mass was removed by ligation with surgical sutures. Histopathological examination revealed conspicuous foci with a variable number of ganglion cells in the tumor and abundant interwoven bundles of schwannian cells with fine fibers. The ganglion cells were positive for neuron-specific enolase and neurofilament. The schwannian cells were positive for vimentin, S-100 protein, and glial fibrillary acidic protein. Thus, according to the classification of tumor with neuronal cell differentiation, the urinary tumor was diagnosed as a ganglioneuroma.     

Prevalence and distribution patterns of <Emphasis Type="Italic">Sarcocystis</Emphasis> spp. in buffaloes in Beni-Suef,Egypt

This study was performed for the purpose of investigating the prevalence of Sarcocystis spp. in buffaloes in Beni-Suef Governorate, Egypt. Both macroscopic (Sarcocystis fusiformis) and microscopic (Sarcocystis levinei) cysts were recognized, and were differentiated by their morphological features and location in the tissues. Of 379 buffaloes examined in abattoirs in Beni-Suef, 299 were found to be infected, with an overall prevalence of 78.9%. Depending on age, three categorized groups of naturally infected buffaloes were examined: male buffalo calves aged 1.5–2 years, adult females aged 2–5 years, and females older than 5 years. Among these groups, infection rates were 74.5%, 82.3%, and 81.2%, respectively. Organs examined included esophagus, tongue, and heart. Macroscopic cysts were examined by the naked eye through meat inspection in abattoirs, while the pepsin-digestion method and the histological technique were applied to detect microscopic cysts. It has been found that esophagus showed the highest rate of infection among the infected organs, with both macroscopic and microscopic cysts seen in the infected buffaloes. Moreover, results of the pepsin-digestion method proved more accurate than those produced by the histological technique in terms of infection rates for the microscopic cysts. Our findings indicated that infected buffaloes aged 2–5 years showed the highest mixed infection rate (82.3%) for both types of cysts. The high prevalence of microscopic Sarcocystis spp. in Beni-Suef Governorate reflects a significant role played by stray dogs, rather than cats, in the transmission of these parasites.