Investigations on the radioactive contamination of crop plants as a result of H-bomb detonation (Part 2) Root and foliage uptake of “Bikini ash”

The Foliar Uptake by Squash Plant

The radioactive ash for experimental use, hereafter referred to as “Bikini ash”, was prepared by igniting the heavily contaminated substances on board No. 5 Fukuryu Maru at about 650°C, followed by sifting through a 100 mesh sieve. On ignition some parts of the fission products, particularly iodine, ruthenium and tellurium would have possibly been lost to the air.     

Sericin Accelerates the Production of Hyaluronan and Decreases the Incidence of Polyspermy Fertilization in Bovine Oocytes During In Vitro Maturation

Fetal bovine serum (FBS) has been widely used as a supplement in the maturation medium of bovine oocytes in vitro. However, serum contains many undefined factors and is potentially infectious to humans and animals. As a serum replacement, we evaluated the feasibility of using the silk protein, sericin, derived from the cocoons of silkworm. To examine the rates of oocyte maturation and fertilization, cumulus-oocyte complexes were cultured in TCM-199 supplemented with 0.01%, 0.05%, 0.1% or 0.15% sericin or 5% FBS. The sizes of the perivitelline space that might relate to polyspermy, the expressions of Has2 and CD44 mRNA, the amount of hyaluronan (hyaluronic acid: HA) contained in the oocytes and the rates of blastocyst formation following insemination were then compared between the oocytes cultured with 0.05% sericin and 5% FBS, because the polyspermy rates in oocytes cultured with 0.05% sericin were significantly lower than in those cultured with 5% FBS. After in vitro maturation (IVM), the mean size of the perivitelline space was significantly greater in oocytes cultured with sericin than in those cultured with FBS, although the rates of nuclear maturation, fertilization and blastocyst formation of oocytes under both IVM conditions were not significantly different. The expression of HAS2 and CD44 mRNA and the amount of HA in the denuded oocytes cultured with 0.05% sericin were significantly greater than in those cultured with FBS. These results indicate the feasibility of sericin as an alternative protein supplement for IVM in bovine oocytes.     

Analysis of Phytosterol,Fatty Acid,and Carotenoid Composition of 19 Microalgae and 6 Bivalve Species

ABSTRACT

In this study, the functional components of 19 microalgae and 6 bivalve species were investigated in the context of the application in bivalve feeding and human health food. Principal component analysis was performed to detect any association between the functional components and individual microalgal species or taxonomic group. The proportions of the functional components differed depending on the taxonomic group or species of microalga. The genera Cheatoceros, Thalassiosira, and Isochrysis contained high concentrations of fucosterol and fucoxanthin, which are present in large amounts in brown algae. Diatoms, haptophytes, and eustigmatophytes, which are used as feed for bivalves, were rich in fucosterol and eicosapentenoic acid (EPA); further, haptophytes were rich in docosahexaenoic acid (DHA). In addition, the microalgae associated with red tide, i.e., the raphidophytes, were found to be rich in fucoxanthin, β-sitosterol, and EPA, whereas dinoflagellates were rich in DHA. Seven bivalve species also contained high concentrations of fucosterol, EPA, and DHA, as did microalgae, which were used to feed by bivalves. These results are useful in selecting microalgae effectively as feed of the bivalves.     

Studies on the physiological effect of humic acid

Many investigations have been reported on the physiological mechanisms of iron chlorosis induced by manganese. And several hypotheses such as “Oxidation reduction potential theory” (1) or “Chelation theory” (2) have been proposed concerning the interference of manganese on the absorption and translocation of iron in plants. However, from a comparative check of experimental results which had been done on various kinds of heavy metals inducing iron chlorosis, the authors have found that the degree of iron chlorosis did not strictly coincide with the order of oxidation reduction potential or stability constant of these chelate compounds. Also, CHINO et al. (3,4) reported that iron chlorosis induced by heavy metals did not coincide with the order of stability constant of these metal chelate compounds, but depended upon their inhibiting action on plant growth and iron absorption. But ASO et al. (5) reported previously that using root separating method they obtained higher iron absorption and better recovery of iron chlorosis of barley plants when iron and nitro-humic acid or EDTA were added simultaneously than when iron only was added. From these results, it was considered that the distribution and chemical form of iron in plants were affected by chelating action of nitro-humic acid or EDTA.     

Changes in the activities of hydroxysteroid dehydrogenases in mouse oocytes during meiotic maturation

The activities of hydroxysteroid dehydrogenases (HSDs) were histochemically demonstrated in mouse oocytes in the process of maturation in vivo and in vitro, and the changes in steroid metabolism during meiotic maturation and also the relationship between nuclear maturation and changes in steroid metabolism in the cytoplasm were examined. In mouse oocytes 0 h after human chorionic gonadotrophin (hCG) injection, the activities of Delta(5)-3beta-HSD (with DHA, pregnenolone and 17alpha-hydroxypregnenolone as the substrates), 17beta-HSD (estradiol-17beta and testosterone) and 20beta-HSD (17alpha-hydroxyprogesterone and 20beta-hydroxyprogesterone) were observed in 87 to 97% of those, but that of 20alpha-HSD (20alpha-hydroxyprogesterone) was not. The percentages of oocytes showing the activities of Delta(5)-3beta-HSD, 17beta-HSD and 20beta-HSD did not change during maturation in vivo or in vitro. Oocytes with 20alpha-HSD activity appeared 4 h after the hCG injection or after culture for 4 h and the rates of those reached 92 and 100%, respectively, 14 h after the hCG injection or after culture for 14 h. In oocytes cultured for 8 h with olomoucine or 3-isobutyl-1-methylxanthine, nuclei were almost all in the germinal vesicle stage, and activity of 20alpha-HSD was observed in 84 and 89% of the treated oocytes, respectively. On the other hand, 81% of control oocytes showed 20alpha-HSD activity, with no significant difference from the rate for the olomoucine- or 3-isobutyl-1-methylxanthine-treated oocytes. The present findings suggested that the metabolic abilities of progesterone, 17alpha-hydroxyprogesterone, 17alpha,20beta-dihydroxyprogesterone, 20beta-hydroxyprogesterone, androgen and estradiol-17beta in the cytoplasm are constantly present in mouse oocytes in the process of maturation in vivo and in vitro. The results also suggested that the metabolic ability of 20alpha-hydroxyprogesterone in mouse oocytes increases during maturation, but the change in the metabolic ability of such a steroid is not related to nuclear maturation.     

Time-lapse videomicrographic analyses of contractions in mouse blastocysts

Contraction has been observed in cultured blastocysts of many mammals, but little is known about the features of the contraction and its physiological role in blastocysts. The author analyzed contractions of a large number of cultured mouse blastocysts by time-lapse videomicrography. The results revealed that blastocysts repeated contractions of different degrees during the expanded stage from 10 h after blastocoel formation, and that the number of contractions was greater during the hatching period than in the periods pre- and post-hatching. The results also showed that the time needed for both contraction and re-expansion to the size before contraction tended to lengthen in blastocysts severely contracted. It was inferred that contractions of blastocysts occur physiologically in relation to myosin light chain kinase, but not due to an increase in permeability between trophectoderm cells in association with their division, or the influence of culture. Furthermore, it was inferred that re-expansion of contracted blastocysts occurs due to active transport and accumulation of Na(+) from the trophectoderm cells into blastocoelic fluid as a result of the action of Na(+)/K(+)-ATPase activated in the membrane of trophectoderm cells. Our results suggested that contractions are also present in blastocysts developed in vivo, and that weak contractions (less than 20% volume reduction) play an important role in hatching, whereas strong contractions (20% or more volume reduction) have the effect of inhibiting hatching. From our results on contractions of various blastocysts, it seems possible to evaluate the developmental ability of embryos, i.e. embryo quality, based on contractions of blastocysts.     

Developmental hormonal profiles in rdw rats with congenital hypothyroidism accompanying increased testicular size and infertility in adulthood

Congenital hypothyroid mutant male rdw rats have enlarged testes in adulthood with dwarfism accompanied by infertility. To explain how rdw rats acquire enlarged testes in adulthood, we compared age-matched normal (N) rats at various developmental stages for blood levels of hormones, thyroxine (T4), follicle stimulating hormone (FSH), luteinizing hormone (LH) and testosterone (T), and investigated whether T4 therapy (rdw+T4) from 3 weeks of age (w) until adulthood could induce recovery of fertility in rdw rats, as well as how rdw+T4 affected hormonal patterns. Testes weights of rdw rats were higher than those of N rats at 19 w in adulthood though it was low during development. Serum T4 values in rdw rats were markedly lower than those in N rats but steadily increased up to 19 w. The serum FSH values in rdw rats were lower than those in N rats at all ages, and neither serum LH nor T value was significantly different at any age. The testes weight of rdw+T4 rats was significantly higher than that of N rats at 13 w with recovered growth, and was higher than that of rdw rats at 19 w. When they were mated with proestrous females after 16 w, all females became pregnant and gave birth to a normal number of pups. The T4 and FSH values of rdw+T4 rats were significantly higher than those in rdw rats, but similar to those in N rats in adulthood. The results suggest that even low levels of circulating thyroid hormone (TH) in rdw rats stimulate the development of their testes, probably through Sertoli cells, resulting in the enlarged adult testes without fertility, and that a sufficient circulating TH level from the immature stage plays a pivotal role in restoring mating activity, probably through FSH-mediated action towards adulthood.     

Changes in the amount of proteins, glycogen and lipids in porcine oocytes during in vitro meiotic maturation

Changes in the cytoplasmic inclusions during meiotic maturation were histochemically examined in cultured porcine oocytes. The oocytes contained a small amount of protein and glycogen granules throughout the maturation culture, as well as Sudanophilic lipids composed of small, medium and large droplets. Soon after collection, the amount of Sudanophilic lipid droplets of small and medium size was small and there were 167 ± 11.2 large droplets. After being cultured for 22 h, the number of large lipid droplets decreased remarkably, while the number of small and medium ones increased. There were no differences in the number of Sudanophilic lipid droplets of different sizes between ovulated oocytes and the oocytes cultured for 44 h. The oocytes always contained a large amount of neutral fats and lipoids, but not cholesterols. In the oocytes cultured for 22 h with olomoucine, both the resumption of nuclear maturation and the decrease in the size of the Sudanophilic lipid droplets were inhibited. From the present findings, it appears that the change in the size of the Sudanophilic lipid droplets in the cytoplasm of porcine oocytes is closely related to nuclear maturation.     

Effect of supplemented sericin on the development,cell number,cryosurvival and number of lipid droplets in cultured bovine embryos

Sericin was investigated as an alternative to fetal bovine serum (FBS) for bovine embryo culture. In vitro matured oocytes were developed using 0.05%, 0.1% or 0.15% sericin. The developmental rate, cryosurvival rate and blastulation time of these embryos were compared with those of embryos developed using 5% FBS. The number of lipid droplets was compared among the blastocysts developed using 5% FBS, using 0.05% sericin and in vivo. The rate of cleavage and blastocyst formation was similar among all groups. Blastulation occurred significantly earlier in the embryos developed using 5% FBS than in those developed using sericin at any concentration (P < 0.05). At 72 h after thawing, the cryosurvival rate of the blastocysts developed using 5% FBS and 0.05% sericin were significantly higher compared with those developed using 0.1% and 0.15% sericin (P < 0.05). The blastocysts developed using 0.05% sericin and in vivo produced a significantly fewer number of medium and large lipid droplets than those developed using 5% FBS. These results suggest that the blastocysts developed using 0.05% sericin show characteristics similar to those of the blastocysts developed in vivo and that the use of sericin as an alternative to FBS is feasible.     

Investigations on the radioactive contamination of crop plants as a result of H-bomb detonation (Part 1) Radioactive contamination of crop plants and soils

Since radioactive rain-out covered Japan in March-May 1954, due to the H-bomb detonation at Bikini atoll, the authors started assaying the possibie radioactive contamination of crop plants grown at Tokyo district.