Evaluation of Loads and Sources of Heavy Metals in Tama River, Tokyo

The loads and sources of heavy metals (Cd, Co, Cu, Mn, Mo, Ni, Pb, V, and Zn) in the midstream of Tama River were evaluated on the basis of their chemical analyses in ordinary and stormwater runoff from July 2007 to November 2008. Tama River is one of the three major rivers flowing into Tokyo Bay. The total annual water discharges differed largely for 2007 and 2008, depending on the scales of typhoon rainfalls and other heavy rainfalls in each year. The concentrations of the metals other than Mo in the river did not change markedly at a flow rate of less than approximately 200 m³ s^?1, but at higher flow rates, the concentrations of all the metals increased linearly with the increase in the log of flow rate (r ²?=?0.94–0.99). The annual loads of heavy metals for 2007 and 2008 were estimated using regression equations between the above parameters and the hourly flow rate data for each year. For the metals other than Mo, the contribution of the loads at higher flow rates (>200 m³ s^?1) was much larger than that at lower flow rates (<200 m³ s^?1), showing the importance of the particulate loads (primarily crustal materials) during storm runoff following typhoon rainfalls and other heavy rainfalls. In contrast, the loads of Mo at lower flow rates accounted for major portions (56–78%) of the total loads, because of a relatively small contribution of particulate load during storm runoff. The contribution of the loads of Mo, Zn, Cd, Cu, and Ni at lower flow rates to the annual loads was larger than that of other metals. It was found that the concentrations of these metals in ordinary runoff are strongly affected by the discharge of treated water from sewage treatment plants which are located along the catchment. Thus, treated water from sewage treatment plants may be the primary source contributing to the present pollution of Mo, Zn, Cd, Cu, and Ni in Tokyo Bay.     

Some Measurement Results on Sulfate Aerosol Concentration in and above a Pine Canopy

Profile of sulfate aerosol (SO₄ ^2?) concentration was measured for four days at six heights in and above a 15m-high canopy of pine plantation during 6th and 10th August. 1999. The concentration was the lowest (about 2 nmol/m³) on 6th, and gradually increased to 9th showing the maximum values of about 13 nmol/m³, and then decreased to 2 nmol/m³ on 10th. The vertical profiles of SO₄ ^2? concentration showed mostly higher in the canopy than above the canopy. As for the vertical profiles above the canopy on 8th and 10th, the minimum was observed just above the canopy (16m), showing SO₄ ^2? transport from the upper air layer to the canopy. While on 9th the profiles that are higher concentration just above the canopy and lower at the upper air layer were observed, suggesting SO₄ ^2? emission from the canopy to the upper air layer.     

Structural determination of hypnosin, a spore germination inhibitor of phytopathogenic Streptomyces sp. causing root tumor in melon (Cucumis sp.)

The structure of a germination inhibitor, hypnosin, isolated from phytopathogenic Streptomyces sp. causing root tumor of melon was determined to be 3-acetylaminopyrazine-2-carboxylic acid (1) by mass spectrometry, computational chemical prediction of UV spectrum, and synthesis of candidates. The structure-activity relationship of hypnosin and anthranilic acid was examined, and it was concluded that pyrazinecarboxylic acid or pyridine-2-carboxylic acid was the fundamental structure with activity, that methylation of the carboxyl group or decarboxylation destroyed activity, and that the presence of an amino group was inhibitory to the activity, whereas acetylation or deletion of an amino group enhanced activity. Hypnosin inhibited spore germination of some Streptomyces spp. in addition to the species with which it was isolated.     

Structure and bioactivity of thiosulfinates resulting from suppression of lachrymatory factor synthase in onion

In normal onion (Allium cepa), trans-S-1-propenyl-L-cysteine sulfoxide is transformed via 1-propenesulfenic acid into propanethial S-oxide, a lachrymatory factor, through successive reactions catalyzed by alliinase and lachrymatory factor synthase (LFS). A recent report showed that suppression of the LFS activity caused a dramatic increase in thiosulfinates previously reported as "zwiebelane isomers". After purification by recycle high-performance liquid chromatography and subsequent analyses, we established the planar structure of the putative "zwiebelane isomers" as S-3,4-dimethyl-5-hydroxythiolane-2-yl 1-propenethiosulfinate, in which two of the three molecules of 1-propenesulfenic acid involved in the formation gave the thiolane backbone, and the third molecule gave the thiosulfinate structure. Of at least three stereoisomers observed, one in the (2'R,3'R,4'R,5'R)-configuration was collected as an isolated fraction, and the other isomers were collected as a combined fraction because spontaneous tautomerization prevented further purification. Both fractions showed inhibitory activities against cyclooxygenase-1 and α-glucosidase in vitro.     

Aroma characteristics of stored tobacco cut leaves analyzed by a high vacuum distillation and canister system

An extraction method using a high vacuum distillation extraction apparatus coupled to a canister was newly developed for the analysis and sensory test of tobacco leaf volatiles. We extended the application of the canister that is used in environmental analysis, to the extraction of the aroma components in tobacco leaves. The volatile components with vapor pressures higher than 0.1 mmHg were easily evaporated under decompression and then trapped into the vacuumed canister. After the collection of volatiles, the canister was pressurized by a slow stream of inert gas in order to emit the whole aroma under a controlled flow. Applying a preconcentrator--gas chromatography/mass spectrometry (GC/MS) and sensory test to the headspace gas components, the aroma alteration between 0 and 2 weeks of storage was simultaneously or individually evaluated. As a result, after the storage, alcohols such as 1-hexanol, linalool, and benzyl alcohol decreased significantly. The amount of carotenoid derivatives that have the characteristic tobacco leaf aroma had not changed. Sensory evaluation of the same headspace gas with that used for GC/MS demonstrated the alternation of the aroma quality before and after storage. The main changes were the decrease of greenness and smoothness in aroma and the decrease of ethylbenzene, 2-pentylfuran, 1-hexanol, benzaldehyde, and linalool concentrations.     

Analysis of radiosensitivity of cancer stem‐like cells derived from canine cancer cell lines

Cancer stem‐like cells (CSCs) are self‐renewing cells comprising a small subpopulation in tumours, and generate differentiated progeny through asymmetric division. It has been shown that CSCs are resistant to ionizing radiation, and this feature could be one of the mechanisms of tumour recurrence after radiation therapy. Much attention has been focused on to target CSCs; however, difficult of isolating CSCs and lack of knowledge on their radiosensitivity have limited this kind of research in veterinary medicine. In the present study, sphere‐forming cells (SC), cultured using sphere formation method, were isolated from four type of canine tumour cell lines and evaluated if they have CSCs‐like properties by expression of CSCs markers (real‐time polymerase chain reaction) and capacity of tumorigenesis (xenograft transplantation in nude mice), and were assessed radiosensitivity (clonogenic survival assay) and DNA repair kinetics (immunofluorescence staining for p53‐binding protein 1) after X‐ray irradiation in comparison with the corresponding normal adherent culture cells (AC). All SCs were isolated using sphere formation and showed high gene expression of CD133 and tumorigenic ability as compared with AC. All SCs were significantly resistant against X‐ray irradiation as compared with AC. In addition, the amount of DNA double‐strand breaks after X‐ray irradiation were significantly lower in SC compared with the corresponding AC. These results indicate that SC isolated through sphere formation possess CSCs‐like characteristics and CSCs are important factor that affect radiosensitivity in canine tumours. In addition, radioresistance of CSCs may depend on reaction of DNA double‐strand break after X‐ray exposure.     

Effect of multiple oral dosing of fluconazole on the pharmacokinetics of cyclosporine in healthy beagles

Fluconazole (Fcz) is successfully used in human organ transplant patients as an antifungal therapy. However, Fcz can increase the cyclosporine (CsA) trough level and lead to CsA nephrotoxicity. In canine renal transplantation, CsA has been used as a major immunosuppressant, and it is important to control its trough level. However, the interaction of Fcz with CsA has not yet been reported in dogs. In this study, the effect of Fcz treatment on the pharmacokinetics of CsA in four healthy beagles was investigated using a four-period crossover design. The treatments included CsA alone (A), CsA + multiple-dose Fcz 50 mg (B), CsA + multiple-dose Fcz 25 mg (C) and CsA + single-dose Fcz 50 mg (D). Blood CsA concentrations were measured at 0.5, 1, 2, 4, 6, 8, 10, 12 and 24 hr after CsA administration. The AUC(0-12) and C(max) values for treatment B were significantly higher than those for the other treatments. In particular, the AUC(0-12) of treatment B was about two times higher than that of treatment A. Fcz administration did not significantly prolong the half-life or mean residence time of CsA. The results of our study show that administration of multiple therapeutic doses of Fcz can significantly increase the CsA blood concentration, which might partially depend upon the Fcz blood concentration. When Fcz is used in CsA-based canine renal transplantation, it may be necessary to adjust the CsA trough level by decreasing the dose.     

包膜尿素氮素的溶出特性研究

 【目的】研究包膜尿素（LP 30）氮素的溶出特性。【方法】采用土壤、纯水两种培养方式,在4种温度（10、15、20、25 ℃）条件下对不同培养时期氮素溶出率进行测定,利用具体的方法——The number of days transformed to standard temperature（DTS）,对自然温度条件下氮素的溶出率进行预测。【结果】培养试验表明,土壤和纯水两种培养方式对包膜尿素氮素溶出率的影响很小,不同温度对氮素溶出率的影响显著。分析表明,一级反应动力学模型可很好地描述包膜尿素氮素的溶出过程。氮素溶出速率随温度升高而加快,在10～25℃内,每升高5℃,速率常数约提高1倍左右。包膜尿素埋藏土壤后实测的氮素溶出率值与采用DTS法预测的氮素溶出率值之间无明显差异。【结论】包膜尿素（LP 30）氮素溶出率大小主要取决于温度的变化,DTS法能较准确地预测包膜尿素施入土壤后,自然温度条件下,任意天数的氮素溶出率。     

Influence of intergeneric/interspecies mitochondrial injection; parthenogenetic development of bovine oocytes after injection of mitochondria derived from somatic cells

Interspecies/intergeneric mitochondrial heteroplasmy can occur in interspecies/intergeneric hybrid embryos or following nuclear transfer. In the present study, intergeneric buffalo (Bubalus bubalis) mitochondria (WB-mt) or interspecies murine (Mus spretus) mitochondria (M-mt) were injected into bovine (Bos taurus) oocytes, and the subsequent embryonic development was characterized. Fibroblast mitochondria (WB-mt or M-mt) were microinjected into in vitro matured bovine oocytes followed by oocyte activation by a combination of electrical stimulation and 6-dimethylaminopurine treatment. After seven days of culture, embryo development was evaluated. The copy number of specific mtDNA populations (introduced and native mtDNA) from heteroplasmic oocytes was estimated using real-time PCR. The results illustrated that oocytes injected with either WB-mt or M-mt can develop to the blastocyst stage (20.6% and 19.6%). Cleavage division rates and development to the morula stage in oocytes injected with WB-mt were lower (76.2% and 45.9%, respectively) in comparison with uninjected oocytes (89.2% and 59.1%, respectively) (P<0.05). However, no differences were found in comparing M-mt injected oocytes and controls (P>0.05). An increase in bovine mtDNA copy number was observed at the expanded blastocyst stage of injected embryos (P<0.01), while the number of injected mtDNA was stable throughout development. This study demonstrates that interspecies/intergeneric mitochondrial injected bovine oocytes have the ability to develop to the blastocyst stage after parthenogenetic activation and that injected mtDNA was neither selectively destroyed nor enhanced through development. Moreover, injected intergeneric mitochondria had a demonstrated influence on bovine parthenogenetic development and mtDNA replication.