Diesel exhaust particle toxicity on spermatogenesis in the mouse is aryl hydrocarbon receptor dependent

Diesel exhaust particles (DEPs) are particulate matter from diesel exhaust containing many toxic compounds, such as polyaromatic hydrocarbons (PAHs). Some toxicities of PAH are considered to express via aryl hydrocarbon receptor (AhR). We hypothesized that the male reproductive toxicity of DEPs may depend on PAHs. BALB/c male mice received 24.7, 74.0 or 220 microg/mouse DEP suspension or vehicle injected into the dorsal subcutaneous layer 10 times during 5 weeks. The mice were euthanized, and blood and organs were collected 2 weeks after the last treatment. The epididymis weights, relative epididymis weights per body weight and daily sperm productions and viabilities of the 74.0 and 220 microg/mouse DEP-treated groups decreased significantly compared with those of the vehicle group. The total incidence of sperm abnormalities in the 74.0 and 220 microg/mouse DEP-treated groups increased significantly compared with the vehicle group. The seminiferous epithelium area ratios of the 74.0 and 220 microg/mouse DEP-treated groups were significantly higher compared with the vehicle and 24.6 microg/mouse DEP-treated groups. The ratios of seminiferous tubules with elongated-type spermatids in the 74.0 and 220 microg/mouse DEP-treated groups were significantly decreased compared with the vehicle group. The testosterone level and hepatic ethoxyresorufin-O-deethylase (EROD) activity as an indirect index of AhR activity in the 74.0 microg/mouse DEP-treated group were significantly increased compared with those of the vehicle group. These results clearly demonstrated that DEPs suppress testicular function, especially spermatogenesis and sperm motility. These effects may be AhR dependent.     

Abnormal distribution of chromosomes in the first division of nuclear transferred mouse embryos

The majority of somatic cell nuclear transferred (SCNT) embryos die before or after implantation. Many studies have focused on morphological remodeling of the donor nucleus and its associated cytoskeletal structures in the early events of nuclear transfer. However, little is known about the 2-cell stage of SCNT embryos after the first division. In this study, we compared the morphological status of chromosomal division during the 1-cell stage to the 2-cell stage in SCNT embryos with that in intracytoplasmic sperm injection (ICSI) embryos. The microtubules and cytoplasmic asters, which are related to chromatin segregation, disappeared at the pronuclear stage, although formation of the first mitotic spindle was normal in both the SCNT and ICSI embryos. However, nuclear fragmentation was observed in 30% of the 2-cell SCNT embryos and 12% of the 2-cell ICSI embryos. Nuclear fragmentation was present in both blastomeres of these embryos. No apoptotic DNA fragmentation was observed in TdT-mediated dUTP-biotin Nick End Labeling (TUNEL) assays for either the SCNT or ICSI embryos. In both the SCNT and ICSI embryos, the distribution of chromosomes in the first mitotic spindle was disturbed during the process of division from the 1-cell stage to the 2-cell stage. These results suggest that loss of SCNT embryos just before or after implantation may be due to an abnormal chromosome distribution at the 2-cell stage.     

Effect of phosphorus on zinc toxicity in tea pollen tube growth

Although zinc (Zn) is an essential element for the growth of higher plants, excess supply may lead to growth inhibition. Symptoms of Zn toxicity are characterized by a reduction in root growth and leaf expansion followed by chlorosis (Mengel and Kirkby 1987), especially root elongation is severely inhibited (Godbold et al. 1983). Due to the increase of input of Zn to farmland by the application of sewage sludge or large amounts of Zn-containing pig manure, Zn toxicity may become an important problem in certain regions (Ruano et al. 1987). In addition, Zn toxicity is also a problem in some acidic soils (Takahashi et al. 1980).

Increasing soil pH by liming is the most effective procedure for decreasing both Zn content and Zn toxicity in plants (White et al. 1979), because Zn solubility decreases 100 times for each unit increase in pH (Neue and Lantin 1994). As an alternative approach, application of large amounts of phosphorus (P) fertilizer was employed to detoxify Zn (Takahashi et al. 1980). However, the mechanisms responsible for the detoxification of the excess Zn by P is remained to be elucidated.

Interactions between Zn and P, which may occur in the rhizosphere and in the uptake and translocation processes, are complex. To separate these factors, in the present study, pollen tubes of tea (Camellia sinensis L.) were used as a model, and the efrect of P on pollen tube growth under Zn toxicity was studied.     

H2 15O translocation in rice was enhanced by 10 μm 5-aminolevulinic acid as monitored by positron emitting tracer imaging system (PETIS)

5-Aminolevulinic acid (ALA) acts to increase chlorophyll biosynthesis, photosynthesis, cold stress tolerance, and salt tolerance at low concentrations. We studied the effects of ALA on H₂ ¹⁵O translocation from the roots to the shoots of rice plants (Oryza sativa L. cv. Nipponbare) in real time by a positron-emitting tracer imaging system (PETIS). When the plant was treated with 10 μm ALA, the velocity of the H₂ ¹⁵O translocation from 2 to 12 min after absorption increased to 126, 137, 140% that of the control at 1.5, 2.5, and 3.5 h after ALA treatment, respectively. However, ALA did not affect the H₂ ¹⁵O translocation within 0.5 h of treatment. When the plant was treated with 0.1 mM ABA at 4 h after 10 μm ALA treatment, the velocity of the H₂ ¹⁵O translocation decreased at 0.5 h after ABA treatment. Those observations suggested ALA might be absorbed and transported to the guard cells within 1.5 h and functioned to expand the stomatal aperture.     

Inhibitory effect of monogalactosyldiacylglycerol, extracted from spinach using supercritical CO2, on mammalian DNA polymerase activity

We investigated the effective extraction of monogalactosyldiacylglycerol (MGDG) from dried spinach (Spinacia oleracea) using supercritical fluid carbon dioxide (SC-CO(2)) with a modifier/entrainer. The yield of MGDG in the SC-CO(2) extract was not influenced by increasing temperature at a constant pressure, although the total extract yield was decreased. The total extract yield and MGDG yield in the extract from commercially purchased spinach (unknown subspecies), were greatly influenced by lower pressure. In a modifier (i.e., ethanol) concentration range of 2.5-20%, both the extract and MGDG yield increased as the ethanol concentration rose. The highest total extract yield (69.5 mg/g of spinach) and a good MGDG yield (16.3 mg/g of spinach) were obtained at 80 degrees C, 25 MPa, and 20% ethanol. The highest MGDG concentration (76.0% in the extract) was obtained at 80 degrees C, 25 MPa, and 2.5% ethanol, although the total extract yield under these conditions was low (5.2 mg/g of spinach). The optimal conditions for the extraction of MGDG were 80 degrees C, 20 MPa, and 10% ethanol. Of the 11 subspecies of spinach tested under these conditions, "Ujyou" had the highest concentration of MGDG. The total extract yield and MGDG concentration of Ujyou were 20.4 mg of the extract/g of spinach and 70.5%, respectively. The concentration of MGDG was higher in the SC-CO(2) extract than in the extract obtained using solvents such as methanol and n-hexane. The extract of Ujyou, which was the optimal subspecies for the extraction of MGDG, inhibited the activity of calf DNA polymerase alpha with IC(50) values of 145 microg/mL but was not effective against DNA polymerase beta.     

Enterolithiasis in a cat

An 8-year-old female Persian cat was brought in for evaluation of chronic vomiting. The presence of opaque enteric foreign bodies and intestinal obstruction along with azotaemia, hyperphosphataemia, moderate anaemia and peritoneal fluid were revealed following appropriate diagnostic work-up. Exploratory laparotomy confirmed jejunoileal dilation, ileocaecal stenosis, and numerous foreign objects in the jejunoileum. These foreign objects and ileocaecal stenosis were surgically removed, and intestinal resection and anastomosis was performed. The patient recovered favourably. Analysis revealed that the foreign objects were composed of calcium phosphate and calcium carbonate. Intestinal inflammation and stenosis secondary to enterolithiasis may have developed following ingestion of cat litter or a previous unrelated surgical intervention. We were unable to delineate the inciting pathogenesis in this particular case.     

Effect of different microbubble generation methods on growth of Japanese mustard spinach

To provide the necessary oxygen for plant growth, the effectiveness of different generating methods of microbubbles (MB), a gas-water circulation type or a pressurization type (pressurization treatment), was investigated by comparing the growth characteristics and morphological observation of Brassica campestris L. var. Komatsuna Matsum. The plant growth in the pressurization treatment were significantly less than those in the circulation treatment at 3 weeks after planting. In plants grown in the pressurization treatment, epidermal cell abrasion and plasmolysis, indicators of cell death, were observed in root tip cells at 3 weeks after planting. The growth inhibition of plant in the pressurization treatment is suggested to be related to the oxidization of Fe in the nutrient solution under super-saturated dissolved oxygen conditions, the generation of hydroxyl radicals from MB, the oxidization of the root tip cell membrane, and osmotic stress to the roots.     

Stomach Chitinase from Japanese Sardine Sardinops melanostictus: Purification,Characterization, and Molecular Cloning of Chitinase Isozymes with a Long Linker

Fish express two different chitinases, acidic fish chitinase-1 (AFCase-1) and acidic fish chitinase-2 (AFCase-2), in the stomach. AFCase-1 and AFCase-2 have different degradation patterns, as fish efficiently degrade chitin ingested as food. For a comparison with the enzymatic properties and the primary structures of chitinase isozymes obtained previously from the stomach of demersal fish, in this study, we purified chitinase isozymes from the stomach of Japanese sardine Sardinops melanostictus, a surface fish that feeds on plankton, characterized the properties of these isozymes, and cloned the cDNAs encoding chitinases. We also predicted 3D structure models using the primary structures of S. melanostictus stomach chitinases. Two chitinase isozymes, SmeChiA (45 kDa) and SmeChiB (56 kDa), were purified from the stomach of S. melanostictus. Moreover, two cDNAs, SmeChi-1 encoding SmeChiA, and SmeChi-2 encoding SmeChiB were cloned. The linker regions of the deduced amino acid sequences of SmeChi-1 and SmeChi-2 (SmeChi-1 and SmeChi-2) are the longest among the fish stomach chitinases. In the cleavage pattern groups toward short substrates and the phylogenetic tree analysis, SmeChi-1 and SmeChi-2 were classified into AFCase-1 and AFCase-2, respectively. SmeChi-1 and SmeChi-2 had catalytic domains that consisted of a TIM-barrel (β/α)₈–fold structure and a deep substrate-binding cleft. This is the first study showing the 3D structure models of fish stomach chitinases.