Spindle formation and microtubule organization during first division in reconstructed rat embryos produced by somatic cell nuclear transfer

The present study was conducted to demonstrate the spindle formation and behavior of chromosomes and microtubules during first division in reconstructed rat embryos produced by somatic cell nuclear transfer (SCNT) with cumulus cell nuclei. To demonstrate the effect of oocyte aging after ovulation on the cleavage of SCNT embryos, micromanipulation was carried out 11, 15 and 18 h after injection of hCG. SCNT oocytes were activated by incubation in culture medium supplemented with 5 microM ionomycin for 5 min followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) in mR1ECM for 2-3 h. For immunocytochemical observation, the SCNT embryos were incubated with monoclonal anti-alpha-tubulin antibody and then fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Cleavage rates were significantly higher for oocytes collected after 15 and 18 h rather than for those collected 11 h after injection of hCG (56 and 53%, respectively vs. 28%; P<0.05). Premature chromosome condensation occurred before activation of the SCNT oocytes, but adequate spindle formation was only rarely observed. The distribution of microtubules in SCNT embryos after activation was different from those of fertilized and parthenogenic oocytes, i.e., a dense microtubule organization shaped like a ring was observed. Eighteen to 20 h post-activation, most SCNT embryos were in the 2-cell stage, but no nucleoli were clearly visible, which was quite different from the fertilized oocytes. In addition, first division with and without small cellular bodies containing DNA was observed in the rat SCNT embryos in some cases. The present study suggests that reorganization of transferred nuclei in rat SCNT embryos may be inadequate in terms of formation of the mitotic assembly and nucleolar reorganization.     

Performance of agency-managed and farmer-managed irrigation systems: A comparative case study at Chitwan, Nepal

Management reforms are considered one of the best alternatives in increasing efficiency of the irrigation systems. Transfer of day-to-day operation and maintenance to farmers is not a new concept. However due to lags, even now it is a hot issue in developing countries and Nepal is not an exception. In this context of growing argument about the transfer of management of public irrigation systems to the Water Users' Group, this paper examines the comparative performance of farmer-managed and agency-managed irrigation systems. Khageri Irrigation System in the Chitwan district of Nepal was taken as the study site where complete management transfer has been done in 1996. The situations before and after complete transfer were compared. Intensive case studies of the selected farmers showed that equity in distribution of water and leakage had significantly improved after management transfer. Logit analysis revealed that age and education level of the household head, distance from main canal, leakage of the canal and equity in distribution are significantly related to the satisfaction level of the beneficiary farmers from FMIS compared to AMIS. However, landholding size of the respondent farmer has no significant effect. Similarly results from t-test revealed that rice productivity and overall profits from agriculture has also increased significantly due to irrigation management transfer.     

Effects of tris(1,3-dichloro-2-propyl) phosphate on epididymal sperm parameters in adult male rats

Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) is widely used as a flame retardant and is known to exhibit anti-androgenic effects in vitro and in vivo. To assess the reproductive toxicity potency of TDCIPP, we investigated the effects of 7 days of TDCIPP oral administration on epididymal sperm motion and concentration in adult male Wistar–Imamichi rats. Thirty-five days after the final administration, sperm parameters were evaluated by computer-assisted sperm analysis. Results showed that sperm swimming progression and vigor and sperm concentration in TDCIPP-treated rats were unexpectedly higher than those in control rats. TDCIPP did not significantly affect the percentage of motile sperms or sperm swimming pattern. These results contribute to the understanding of the biological effects of TDCIPP.     

Evaluation of Intestinal Absorption of Dietary Halocynthiaxanthin,a Carotenoid from the Sea Squirt Halocynthia roretzi

Halocynthiaxanthin is an acetylenic carotenoid mainly found in Halocynthia roretzi. To date, several bioactivities of halocynthiaxanthin have been reported, but its mechanism of digestion and absorption in mammals has not been studied yet. In this study, we evaluated the intestinal absorption of halocynthiaxanthin in mice. The halocynthiaxanthin-rich fraction was prepared from the tunicate Halocynthia roretzi. Mice were orally administered the fraction at a dose of 5 mg/kg body weight. The halocynthiaxanthin levels in the plasma, liver, and small intestine, were quantified using HPLC-PDA, 1, 3, 6, and 9 h after ingestion. The halocynthiaxanthin-rich fraction mainly consisted of the all-trans form and a small amount of cis forms. These three isomers were detected in the plasma of mice 3 h after ingestion. Time-course changes after the ingestion of this fraction were found, with cis isomers being more abundant than the all-trans isomer in the mouse plasma and liver. In the small intestine, however, the all-trans isomer was primarily detected. The possibility that cis isomers might be absorbed rapidly from the small intestine cannot be denied, but our results suggest that dietary all-trans-halocynthiaxanthin might be isomerized to the cis isomer after intestinal absorption.     

Development and testing of a surface roughness measurement device based on aerial ultrasonic reflections

Paddy and Water Environment - Repairs of concrete irrigation channels in Japan are guided to a large extent by the degree to which their walls have degraded over decades of use. Current methods of...     

Chemical and Thermal Properties of Freshwater Prawn (Macrobrachium rosenbergii) Meat

Chemical compositions and thermal properties of cultured freshwater prawn meat (FPM) were studied. FPM contained 83.2% protein (dry basis), 62.7% of which was myofibrillar protein. Pepsin-soluble collagen (PSC) and insoluble collagen (ISC) contents were 0.63 and 0.32%, respectively. Both collagens were similar to type V collagen from porcine placenta. Glutamic acid/glutamine, arginine, aspartic acid/?asparagine, and lysine were abundant amino acids in FPM. Glycine, proline, hydroxyproline, and aspartic acid/?asparagine were predominant in both collagens. FPM exhibited thermal transition temperatures (T_max) of 48.3 and 64.7°C, whereas T_max of PSC and ISC were 43.0 and 46.0°C, respectively. Textural changes in FPM during post-mortem storage on ice are plausibly dependent upon its compositional and thermal properties.     

Genetic variability and stock structure of red tilefish Branchiostegus japonicus inferred from mtDNA sequence analysis

Sequence analysis based on the anterior part of the mitochondrial DNA control region was carried out to reveal the genetic diversity, stock structure, and historical demography of the red tilefish. Nucleotide sequences of 388 bp in length were determined for 280 individuals taken from eight localities. The molecular diversity, haplotype diversity, and nucleotide diversity were relatively high (average h = 0.929 ± 0.011; average π = 0.008 ± 0.005). The mismatch distribution was not significantly different from the expected distribution for a rapidly expanding population (P = 0.453). The minimum spanning network connecting with full-sequence haplotypes contained star-like topologies derived from multiple ancient lineages, supporting the mismatch distribution analysis. No significant genetic differentiation was observed among eight localities (maximum pairwise F _ST = 0.023, reduced-sequence data set). Our results suggest a large panmictic population of the red tilefish along the coast of Honshu to the East China Sea.     

Binding profiles and cytokine-inducing effects of fish rhamnose-binding lectins on Burkitt’s lymphoma Raji cells

Rhamnose-binding lectin (RBL) is one of the animal lectin categories which take part in the innate immune responses of fish. Osmerus lanceolatus lectin (OLL) from shishamo smelt eggs is an RBL composed of two tandem-repeated domains, both of which are considered to be a carbohydrate-recognition domain. SAL, catfish (Silurus asotus) egg RBL composed of three domains, binds to Burkitt’s lymphoma Raji cells through globotriaosylceramide (Gb3) carbohydrate chain and to reduce cell size and growth by altering membrane composition without causing cell death. In this experiment, we tried to compare the binding effects of these two RBLs on Raji cells. Flow cytometric and fluorescence microscopic analyses revealed that OLL also directly bound to and shrunk Raji cells with ten times less reactivity than SAL but reduced cell growth with decreasing cell viability. Anti-Gb3 antibody completely blocked the binding of SAL to Raji cells but not that of OLL. In addition, the direct bindings of OLL and SAL to Raji cells were comparably inhibited by melibiose, but lactose was more effective inhibitor for the binding of OLL than that of SAL. These results suggest that OLL has slightly different cell-binding property compared with SAL and binds not only to Gb3 but also to the other carbohydrate receptor-bearing β-galactoside chains. The quantitative RT-PCR analysis revealed that SAL induced the expression of TNF-α but not of IFN-γ, IL-1β, and IL-10. Thus, SAL-induced cytostatic effect on Raji cells might be partially caused by TNF-α-mediated signaling pathway.     

Varietal differences in phenolic content and astringency in skin and flesh of hardy kiwifruit resources in Japan

Varietal differences in the total phenolic content and astringency in the skin and flesh were determined among the cultivars and local collections of hardy kiwifruit with a ploidy variance found in Japan. The average values of the total phenolic content in the skin and flesh were 2.66 and 0.18 g 100 g⁻¹ FW, respectively. There were large varietal differences in the total phenolic content in the skin in the range of 1.3–5.0 g 100 g⁻¹ FW. Kochi (tetraploid), while Gassan and ‘Mitsuko’ (hexaploid) contained a larger amount of total phenolics. High astringency was found in Gassan, ‘Mitsuko’ and ‘Hoko’ (hexaploids) and Kochi. HPLC analysis showed that the major components of phenolics in the flesh were (+)-catechin, chlorogenic acid, rutin, (−)-epicatechin and quercetin.     

Influence of volcanic ash sediments on spore adhesion of the red alga Gelidium elegans from the coastal seabed around Miyakejima Island, Japan

The populations of red alga Gelidium elegans along the coast of Miyakejima Island were severely damaged by a volcanic eruption in 2000. The effect of this volcanic eruption has been long lasting, and populations of this red alga still have not recovered. We investigated the effect of seabed sediment particles derived from volcanic ash on the substrate adhesion of G.?elegans spores. The analysis provides evidence that increasing amounts of sediment particles result in lower adhesion rates of G.?elegans spores, and that smaller sediment particles have a greater influence on adhesion. The amount of seabed sediment particles around Miyakejima Island was 9.3?C1815.4?mg/cm². This amount has changed greatly from year to year. The adhesion rate of G.?elegans spores in water around Miyakejima Island was 0?% at all points in 2008 and 2010, but it was estimated as 6.3?C38.6?% in 2009. These results suggest that there is significant inhibition of algal spore adhesion by seabed sediment particles derived from volcanic ash around Miyakejima Island.