Epigenetic assessment of environmental chemicals detected in maternal peripheral and cord blood samples

Epigenetic alteration is an emerging paradigm underlying the long-term effects of chemicals on gene functions. Various chemicals, including organophosphate insecticides and heavy metals, have been detected in the human fetal environment. Epigenetics by DNA methylation and histone modifications, through dynamic chromatin remodeling, is a mechanism for genome stability and gene functions. To investigate whether such environmental chemicals may cause epigenetic alterations, we studied the effects of selected chemicals on morphological changes in heterochromatin and DNA methylation status in mouse ES cells (ESCs). Twenty-five chemicals, including organophosphate insecticides, heavy metals and their metabolites, were assessed for their effect on the epigenetic status of mouse ESCs by monitoring heterochromatin stained with 4￠,6-diamino-2-phenylindole (DAPI). The cells were surveyed after 48 or 96 h of exposure to the chemicals at the serum concentrations of cord blood. The candidates for epigenetic mutagens were examined for the effect on DNA methylation at genic regions. Of the 25 chemicals, five chemicals (diethyl phosphate (DEP), mercury (Hg), cotinine, selenium (Se) and octachlorodipropyl ether (S-421)) caused alterations in nuclear staining, suggesting that they affected heterochromatin conditions. Hg and Se caused aberrant DNA methylation at gene loci. Furthermore, DEP at 0.1 ppb caused irreversible heterochromatin changes in ESCs, and DEP-, Hg- and S-421-exposed cells also exhibited impaired formation of the embryoid body (EB), which is an in vitro model for early embryos. We established a system for assessment of epigenetic mutagens. We identified environmental chemicals that could have effects on the human fetus epigenetic status.     

How different or similar are nematode communities between a paddy and an upland rice fields across a flooding-drainage cycle?

The paddy field is being recognised as a biodiversity hotspot fostering a variety of organisms. However, there are few studies on the ecology of paddy field nematodes. We characterised nematode communities in rice paddy fields by comparing them with upland fields of rice or soybean. We examined nematode communities of the top (0-15 mm) and second (15-50 mm) soil layers before flooding (March or April), during flooding (June or July) and during the draining period (October) 2007-2009. We found that the nematode community in the paddy was different than that in the upland fields during all periods. Rhabdolaimus, Tobrilus, Mesodorylaims and Monhysteridae characterised the top of the paddy and Hirschmanniella characterised the second layer of the paddy. Total nematode density was generally lower in the paddy than in the upland field. However, the density in the paddy top layer increased with time from the flooding period to the draining period, during which time it was about the same as (or even greater than) the peak density in the upland fields. The density in the second layer of the paddy remained lower than that in the top layer of the paddy throughout the time course. Community diversity values were generally greater in the paddy top layer than in the paddy second layer across the six sampling periods, but periodic differences between the paddy and upland fields or between soil layers were not significant. During the flooding period, the F/(F + B) (13-37) and Enrichment Index (17-38) values were lower in the paddy than in the upland fields (32-47, 37-74, respectively) to reflect that bacteria dominate over fungi with slow decomposition due to anaerobic conditions in the flooded paddy field. In addition, particularly in the top layer, the Maturity (2.0-2.4) and Structure Index (23-72) values were greater in the paddy than in the upland fields (1.7-2.1, 9-15, respectively), indicating a well-developed ecosystem under water. These unique nematode communities persisted during the draining period, but there was a rapid increase in opportunistic bacterivores, which increased the EI values. We suggest that bactivorous nematodes in the families Cephalobidae and Chronogasteridae, herbivores in the genus Hirschmanniella, and fungivores in the genus Filenchus may be specific to paddy field soil rather than to pond and lake sediments.     

Microbial mineralization of organic nitrogen into nitrate to allow the use of organic fertilizer in hydroponics

Hydroponics is an excellent technique for the cultivation of vegetable crops and other plants, but organic fertilizers cannot be used in conventional hydroponic systems, which generally use only inorganic fertilizers, because organic compounds in the hydroponic solutions generally have phytotoxic effects that lead to poor plant growth. Few microorganisms are present in hydroponic solutions to mineralize the organic compounds into inorganic nutrients. In this article a novel and practical hydroponic culture method that uses microorganisms to degrade organic fertilizer in the hydroponic solution has been developed. Soil microorganisms were cultured by regulating the amounts of organic fertilizer and inoculum, with moderate aeration. The microorganisms mineralized organic nitrogen via ammonification and nitrification into nitrate at an efficiency of 97.6%. The culture solution containing the microorganisms was usable as a hydroponic solution, and organic fertilizer could be directly added to it during vegetable cultivation. Vegetables grew well in the organic hydroponic system. Organic hydroponics based on this method is therefore a practical tool for the utilization of organic sources of fertilizer.     

Physiological and behavioural effects of an intracerebroventricular injection of corticotropin releasing hormone in goats

This study investigated the effects of an intracerebroventricular (ICV) injection of corticotropin releasing hormone (CRH) on physiological and behavioural responses in goats. In Experiment 1, saline (control) or saline plus 25 microg of ovine CRH was injected into the third ventricle of castrated male goats. CRH increased plasma cortisol (Cor) levels markedly within 15 min, but had little effect on plasma glucose (Glu). Compared with saline injected goats, CRH decreased the total duration of lying behaviour but increased its frequency, and suppressed rumination and self-grooming. In Experiment 2, the effects of an intravenous (IV) injection of human adrenocorticotropic hormone (ACTH) (1-24) (0.1mg) were examined and an IV injection of saline was used as control. ACTH increased plasma Cor levels markedly, but did not change any behaviour compared with controls. It was concluded that CRH mediated the response of the hypothalamus-pituitary-adrenal (HPA) axis and behaviour following stress in goats, although the CRH-induced behavioural changes were independent of the HPA axis and seemed to be the result of direct action within the central nervous system.     

Removal of heavy metal ions from solutions by coniferous barks

Summary The abilities of 15 coniferous barks for removing toxic heavy metal ions were investigated. The barks considerably varied in the adsorption ability to each metal ion. Of the barks tested, high adsorption ability for heavy metal ions was found inPicea abies (Norway spruce). Equilibrium experiments usingP. jezoensis (Yezo spruce) bark showed that the adsorption of Cd²⁺ was greatly affected by the pH of solution and the initial Cd²⁺ concentration in solution. The adsorption of Cd²⁺ byP. jezoensis bark followed Freundlich isotherm in the concentration range 10–330 mg Cd²⁺/L. The continuous column experiments usingP. jezoensis bark indicated that the packing had retained 10.1–14.2 mg Cd²⁺/g adsorbent until the column broke through.     

The smell and odorous components of dried shiitake mushroom, Lentinula edodes III: substances that increase the odorous compound content

This study formed part of an effort to improve the quality of dried shiitake mushroom [Lentinula edodes (Berk.) Pegler], in accordance with consumer preferences, and deals with the search for substances that increase the odorous component content. From analysis of sulfur and sulfur-containing substances in the culture substrate, rice bran was found to be the main source of sulfur, and 75% of this was present as cysteine and methionine. The sulfur-containing substances were added to a sawdust medium containing only rice bran and sawdust as a substrate; shiitake mushrooms were cultivated in the medium, the fruiting bodies were dried, and the 1,2,4-trithiolane content in the fruiting bodies was measured as an indicator of the odorous compounds. Of the sulfur-containing substances, those that increased odorous compounds the most were cysteine and methionine. The efficiency of cysteine in this regard was higher than that of methionine. It was also noted that the amount of glutamic acid increased the odorous compounds in combination with cysteine and methionine. Furthermore, the addition of both amino acids and glutamic acid had no negative effect on the yield. These results showed that it is possible to produce dried shiitake mushrooms with a smell suitable for particular consumer preferences.     

The impact of forest clear-cutting on soil temperature: a comparison between before and after cutting,and between clear-cut and control sites

Soil temperature is one of the most important factors governing biological activity in the soil. This study was conducted to investigate how forest clear-cutting changes soil temperature. Soil temperatures at 0.5, 1.0, 2.0, and 3.0m depths were measured in two neighboring forest watersheds (35°12N, 140°06E) in Chiba Prefecture, Japan, from 1994 to 2000. One watershed was clear-cut 5 years after the observations began. After clear-cutting, the annual mean soil temperature at 0.5, 1.0, 2.0, and 3.0m depths rose by about 2.2, 2.0, 1.7, and 1.4°C, respectively. The maximum respective soil temperatures rose by about 3.2, 3.0, 2.2, and 1.8°C. The minimum soil temperature rose slightly (1°C). The range of temperatures increased by 3.0, 2.4, 1.6, and 1.4°C, respectively. In our study, forest clear-cutting raised maximum and average soil temperatures but hardly changed minimum soil temperature. This is probably because solar radiation dominated in the summer season and increased soil temperature; on the other hand, net long-wave radiation, and releases of latent and sensible heat from the soil surface, were predominant in the cool season.     

The smell and odorous components of dried shiitake mushroom, <Emphasis Type="Italic">Lentinula edodes</Emphasis> I: relationship between sensory evaluations and amounts of odorous components

The smell of food is one of the most important factors in assessing its quality. Concerning the smell of dried shiitake mushrooms [Lentinula edodes (Berk.) Pegler], 1,2,3,5,6-pentathiepane, commonly known as lenthionine, has been reported as a key compound. However, other compounds have not been studied sufficiently in connection with smell. From the results of sensory intensity studies and sensory evaluations of dried shiitake mushrooms, a positive significant correlation at 1% risk was observed between sensory intensity and sulfur perception. This showed that the smell of dried shiitake mushrooms was characterized by a sulfurous smell. Also, comparing the sensory intensity with the amounts of volatile components showed positive significant correlations at 1% risk between sensory intensity and three compounds: 1,2,4-trithiolane, 1,2,4,6-tetrathiepane, and lenthionine. Furthermore, significant correlations at 5% risk were obtained between the amounts of these three compounds and sensory intensity by multiple regression analysis. This showed that the smell of dried shiitake mushrooms depended on these compounds. The partial regression coefficient of 1,2,4-trithiolane was larger than those of the others, and so it was proposed that 1,2,4-trithiolane could serve as an indicator to estimate the smell of dried shiitake mushroom.     

Adsorption of mercury by sugi wood carbonized at 1000°C

The ability of sugi wood carbonized at 1000°C to adsorb mercury was examined using aqueous solutions of mercuric chloride. Parameters studied include contact time, pH, adsorption temperature, and initial concentration of mercury in solution. Results showed that sugi wood carbonized at 1000°C could effectively remove mercury from aqueous solutions. The carbonized wood showed high adsorption ability for mercury at a wide pH range (pH 3–9), but its ability drastically decreased at pH 11. Adsorption decreased with increases in adsorption temperatures, indicating that the processes were exothermic in nature. Adsorption was found to follow the Freundlich isotherm model. The adsorption capacity of carbonized sugi wood was comparable to that of commercial activated carbon.