Thraustochytrid as a potential source of carotenoids

Thraustochytrids, marine protists whose dominant genera are Thraustochytrium and Schizochytrium, belong to the kingdom Chromista and are known as an industrial source of DHA. We describe here that thraustochytrid strain KH105, isolated as a DHA producer, also accumulates significant levels of β-carotene and xanthophylls including canthaxanthin and astaxanthin. A4-d cultivation using a medium composed of 10% glucose and less than 0.3% of nitrogen sources in a half-concentration of seawater gave an astaxanthin production up to 6.1 mg/L, and canthaxanthin content reached more than 10 mg/L under conditions where a higher concentration of nitrogen sources (6%) was employed. It might be advantageous in mass production systems for these carotenoids to be extracted readily by simply suspending the cells with organic solvents such as acetone and chloroform. Analyses on the morphological and life history features of the KH105 strain revealed that it belongs to the genus Schizochytrium. This particular species of thraustochytrids is thus considered to be a promising source of xanthophylls as well as DHA for use in the food industry.     

Characteristics of chlorophyll formation of the aerial microalga Coelastrella striolata var. multistriata and its application for environmental biomonitoring

The growth and ammonium uptake of the aerial microalga Coelastrella striolata var. multistriata, which was isolated from the surface of rocks, were characterized in this study. The specific growth rate of the alga was mu=0.3 d(-1), as calculated in the growth logarithmic phase. The algal cells were able to remove almost 100% of the ammonium ions from medium in 5 d, with the removal rate of ammonium-N being 0.4 mg/l/h. It was shown that the alga has a unique ability to be a reddish orange to green color depending on the nitrogen source concentration in the medium. Astaxanthin, adonixanthin, canthaxanthin, and beta-carotene were found in the reddish orange cells of the alga. The assessment of water pollution was attempted using this aerial microalga. When the reddish orange alga was incubated in the experimental medium with added ammonium-, nitrate-, or urea-N as a nitrogen source, an approximately linear relationship existed between the nitrogen concentration and chlorophyll formation. Using the chlorophyll formation of the alga, for example, it was possible to estimate spectrophotometrically the total nitrogen content in water collected from aquatic systems. Biofunctional materials for environmental biomonitoring using photosynthetic microorganisms are called green devices in this study.     

Production and properties of phytase and acid phosphatase from a sake koji mold, Aspergillus oryzae

We identified three types of acid phosphatase (ACP-I, ACP-II, and ACP-III) produced by Aspergillus oryzae in a submerged culture using only phytic acid as the phosphorous substrate. The optimum pH for the activities of the three enzymes was in the range of 4.5 to 5.5. Analysis of the substrate specificities of these enzymes revealed that ACP-I and ACP-III were acid phosphatases, and ACP-II was a phytase. These enzymes were produced during different periods of mycelial growth: ACP-II was produced during the early phase of cultivation (around 24 h), and ACP-I was produced between 24 to 72 h. ACP-III was detected after the production of ACP-I and ACP-II had ceased. The release of phosphate from phytic acid was expected to be due to the cooperative hydrolysis of these enzymes.     

Structural development of natural rubber during uniaxial stretching by in situ wide angle X-ray diffraction using a synchrotron radiation

Structural development of natural rubber during uniaxial stretching was examined by an in situ wide angle X-ray diffraction measurement using a synchrotron. During stretching, the amorphous part showed little change, i.e. an amorphous halo remained clear even at 500% strain. The fraction of induced crystals was very small, though a clear crystalline pattern was observed at 400% strain. Some polymer chains were oriented and crystallized, but most of the chains were not oriented at all in spite of large deformations of the specimen. Only a small amount of polymer chains contributes to the stress and hysteresis loss during elongation.     

Distribution and heterogeneity of mast cells in the human uterus

Mast cells (MCs) are widely distributed in most human tissues. Those cells that contain only tryptase are designated as T-MCs, while those that also contain chymase are referred to as TC-MCs. This study uses immunohistochemical staining for tryptase and chymase to assess the distribution and heterogeneity of these two types of MCs in the human uterus. The greatest number of MCs was found in the inner (i.e. luminal) half of the myometrium, with this area containing approximately equal proportions of T-MCs and TC-MCs. There were fewer MCs in the outer half of the myometrium and the cervix, but the proportion of TC-MCs in both of these areas was substantially higher. In contrast, the endometrium contained significantly fewer MCs, but proportionally more T-MCs. There was no change in the number of MCs between the proliferative and secretory phases of the menstrual cycle; however, there was a significantly lower number in all areas after menopause. Most of the MCs were observed in close association with uterine smooth muscle cells, as well as in the vicinity of fibroblasts and collagen, and it appears they may play an important role in the reconstruction of uterine tissues during the menstrual cycle.     

Neurite outgrowth of PC12 cells is suppressed by wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase

The effects of wortmannin (WT), an inhibitor of phosphatidylinositol (PI) 3-kinase, on differentiation of PC12 cells were analyzed. WT inhibited PI 3-kinase activity of PC12 cells at a concentration of 10(-7) M in vivo and in vitro. Transient inhibition of PI 3-kinase activity at the time of nerve growth factor stimulation had no effect on activation of the ras protein or neurite formation by the cells. However, continuous inhibition of PI 3-kinase blocked differentiation at the step just before neurite formation. When WT was applied to cells growing neurites, elongation of the neurites was stopped at that step. These results suggest that PI 3-kinase may be involved in neurite elongation.     

Human small Maf proteins form heterodimers with CNC family transcription factors and recognize the NF-E2 motif

To protect bone marrow cells from the toxicity of chemotherapy, a multidrug resistant gene or a dihydrofolate reductase gene has been introduced into stem cells. These genes, however, are not capable of conferring refractoriness to alkylating agents (AA), which are some of the most commonly used agents in chemotherapy regimens. In the present study, an attempt was made to endow human stem cell (CD34+ cells) with resistance to cyclophosphamide, a well-known AA, and adriamycin (ADM) by transducing the glutathione-S-transferase pi (GST-pi) gene whose product is thought to detoxify AA by conjugating them with glutathione and to remove a toxic peroxide formed by ADM. The gene transduction was carried out retrovirally with a virus titer of 1 x 10(5) FFU/ml, employing a recombinant fibronectin fragment; transduction efficiency was extremely low without the fragment. Incubation with interleukin-6 and stem cell factor enhanced the expression of fibronectin ligands VLA4 and VLA5 on CD34+ cells. This enhanced expression of VLA4 and VLA5 was considered to facilitate a close contact of the CD34+ cell to the retroviral vector via fibronectin fragments and the subsequent transduction process. The GST-pi gene-transduced CD34+ cells formed almost 3- and 2.5-fold more CFU-GM than neo gene-transduced CD34+ cells in the presence of 2.5 microg/ml of 4-hydroperoxycyclophosphamide (4-HC), an active form of cyclophosphamide, and 30 ng/ml ADM, respectively. The transfectants formed an appreciable number of colonies, even at higher concentrations of these drugs (5.0 microg/ml of 4-HC, 50 ng/ml of ADM) whereas neo gene-transduced or nontransduced CD34+ cells formed no colonies at all, indicating the possibility of selecting out the transfectants by exposing them to these anticancer drugs. Thus, we were able to demonstrate that transduction of the GST-pi gene confers resistance to cyclophosphamide as well as to ADM, and therefore this approach can be applied clinically for high-dose chemotherapy.