• Journal of Life Science
• /
• v.31 no.11
• /
• pp.1028-1036
• /
• 2021

Firefly luciferase (FLuc) can function as an efficient marker in the gene and viral therapies. Nonetheless, its clinical translation has been unaccomplished with the concerns on its exogenous nature and the similarity with human fatty acyl-CoA synthetase. In this study, we aimed to show safety of FLuc by conducting a set of preclinical experiments and a human use. Initially, FLuc permeability across the plasma membrane was investigated by delivering the FLuc-carrying viral vector, OTS-412, or the FLuc recombinant protein. After in vitro infection of OTS-412 into different cancer cell lines, FLuc activity was detected only in the cell lysates, but not in culture media. In addition, recombinant FLuc protein further showed the impermeability against the plasma membrane. Similar result was also observed in the in vivo experiment. After being injected into the VX2 tumor-bearing rabbit, the FLuc exclusively resided within the tumor tissue without being detected in the blood plasma or other organs. Human cancer cell lines originated from various organs were lysed and treated to the FLuc, and none of the human substrates was reactive against the FLuc. As a final step, FLuc recombinant protein was intravenously injected into a human. The luciferase was degraded with the half-life of 20 to 30 minutes in blood, and was untraceable from 1.5 hr after the injection. In addition, the blood plasma was nonresponsive against the fatty acids. Hematological analysis was also comparable between the pre- and post-injection. Altogether, our study collectively demonstrates the safety of the firefly luciferase.

• Journal of Aquaculture
• /
• v.2 no.2
• /
• pp.53-90
• /
• 1989

Feeding proper level of ration matchable with the appetite of fish will enhance production and also prevent waste of food and its consequence, side effects such as pollution of culture medium. To pursue this goal, elaborate studies on dissolved oxygen concentrations- as the major force in inducing appetite and the growth outcome are necessary. The growth of common carp of 67, 200, 400, 600, and 800 gram size groups was studied at oxygen concentrations ranging from 2.0 to 6 mg/$\iota$ in relation to rations from 1 to as many percent of the initial body weight as could be consumed under constant temperature of $25^{\circ}C$. The results from the experiments are summarized as followings; 1. Appetite: The smaller fish exhibited higher degree of appetite than the bigger ones at the same oxygen concentrations. The bigger the fish the less tolerant it was to the lower oxygen thersholds, and the degree of tolerence decreased as ration level increased. 2. Growth : Growth rate (percent per day) increased - unless consumption was suppressed by low oxygen levels- as the ration was increased to maximum. In case of 67 g fish, it reached the highest point of $5.05\%$ / day at $7\%$ ration under 5.0 mg/$\iota$ of oxygen. In case of 200 g fish, the maximum growth rate of $3.75\%$/day appeared at the maximum ration of $6\%$ under 5.5 mg/$\iota$ of oxygen. In 400 g fish, the highest growth of $3.37\%$/day occurred at the maximum ration of $5\%$ and 6.0 mg/$\iota$ of oxygen. In 600 g fish, the highest growth rate of $2.82\%$ /day was at the maximum ration of $4\%$ under 5.5 mg/$\iota$ oxygen. In case of 800g fish, the highest growth rate of $1.95\%$/day was at maximum tested ration of $3\%$ under 5.0 mg/$\iota$ oxygen. 3. Food Conversion Efficiency: Food conversion efficiency ($\%$ dry feed converted into the fish tissue) first increased as the ration was increased, reached maximum at certain food level, then started decreasing with further increase in the ration. The maximum conversion efficiency stood at higher feeding rate for the smaller fish than the larger ones. In case of 67 g fish, the maximum food conversion efficiency was at $4\%$ ration within 3.0-4.0 mg/$\iota$ oxygen. In 200g fish, the maximum efficiency was at $3\%$ ration within 4.0-4.5 mg/$\iota$ oxygen. In 400g fish, the maximum efficiency was at $2\%$ ration within 4.0 - 4.5 mg/$\iota$ oxygen. In 600 and 800g fish, the maximum conversion efficiency shifted to the lowest ration ($1\%$) and lower oxygen ranges. 4. Behaviour: The fish within uncomfortably low oxygen levels exhibited suppressed appetite and movements and were observed to pass feces quicker and in larger quantity than the ones in normal condition; in untolerably low oxygen the fish were lethargic, vomited, and had their normal skin color changed into pale yellow or grey patches. All these processes contributed to reducing food conversion efficiency. On the other hand, the fish within relatively higher oxygen concentrations exhibited higher degree of movement and their food conversion tended to be depressed when compared with sister groups under corresponding size and ration within relatively low oxyen level. 5. Suitability of Oxygen Ranges to Rations: The oxygen level of 2.0- 2.5 mg/$\iota$ was adequate to sustain appetite at $1\%$ ration in all size groups. As the ration was increased higher oxygen was required to sustain the fish appetite and metabolic activity, particularly in larger fish. In 67g fish, the $2\%$ ration was well supported by 2.0-2.5 mg/$\iota$ range; as the ration increased to $5\%$, higher range of 3.0-4.0 mg/$\iota$ brought better appetite and growth; from 5 till $7\%$ (the last tested ration for 67 g fish) oxygen levels over 4.0 mg/$\iota$ could sustain appetite. In 200 g fish, the 2 and $3\%$ rations brought the best growth and conversion rates at 3.5-4.5 mg/$\iota$ oxygen level; from 3 till $6\%$ (the last tested ration at 200 g fish) oxyge groups over 4.5 mg/$\iota$ were matchable with animal's appetite. In 400, 600, and 800 g fish, all the rations above $2\%$ had to be generally supported with oxygen levels above 4.5 mg/$\iota$.

• Journal of Korean Society of Forest Science
• /
• v.21 no.1
• /
• pp.1-34
• /
• 1974

The author intended to investigate external and internal changes in the cone structure, changes in water content, sugar, fat and protein during the period of seed maturation which bears a proper germinability. The experimental results can be summarized as in the following. 1. Male flowers 1) Pollen-mother cells occur as a mass from late in April to early in May, and form pollen tetrads through meiosis early and middle of May. Pollen with simple nucleus reach maturity late in May. 2) Stamen number of a male flower is almost same as the scale number of cone and is 69-102 stamens. One stamen includes 5800-7300 pollen. 3) The shape is round and elliptical, both of a pollen has air-sac with $80-91{\mu}$ in length, and has cuticlar exine and cellulose intine. 4) Pollen germinate in 68 hours at $25^{\circ}C$ with distilled water of pH 6.0, 2% sugar and 0.8% agar. 2. Female flowers 1) Ovuliferous scales grow rapidly in late April, and differentiation of ovules begins early in May. Embryo-sac-mother cells produce pollen tetrads through meiosis in the middle of May, and flower in late May. 2) The pollinated female flowers show repeated divisions of embryo-sac nucleus, and a great number of free nuclei form a mass for overwintering. Morphogenesis of isolation in the mass structure takes place from the middle of March, and that forms albuminous bodies of aivealus in early May. 3. Formation of pollinators and embryos. 1) Archegonia produce archegonial initial cells in the middle and late April, and pollinators are produced in the late April and late in early May. 2) After pollination, Oespore nuclei are seen to divide in the late May forming a layer of suspensor from the diaphragm in early June and in the middle of June. Thus this happens to show 4 pro-embryos. The organ of embryos begins to differentiate 1 pro-embryo and reachs perfect maturation in late August. 4. The growth of cones 1) In the year of flowering, strobiles grow during the period from the middle of June to the middle of July, and do not grow after the middle of August. Strobiles grow 1.6 times more in length 3.3 times short in diameter and about 22 times more weight than those of female flower in the year of flowering. 2) The cones at the adult stage grow 7 times longer in diameter, 12-15 times shorter diameter than those of strobiles after flowering. 3) Cone has 96-133 scales with the ratio of scale to be 69-80% and the length of cone is 11-13cm. Diameter is 5-8cm with 160-190g weight, and the seed number of it is 90-150 having empty seed ratio of 8-15%. 5. Formation of seed-coats 1) The layers of outer seed-coat become most for the width of $703{\mu}$ in the middle of July. At the adult stage of seed, it becomes $550-580{\mu}$ in size by decreasing moisture content. Then a horny and the cortical tissue of outer coats become differentiated. 2) The outer seed-coat of mature seeds forms epidermal cells of 3-4 layers and the stone cells of 16-21 layers. The interior part of it becomes parenchyma layer of 1 or 2 rows. 3) Inner seed-coat is formed 2 months earlier than the outer seed-coat in the middle of May, having the most width of inner seed-coat $667{\mu}$. At the adult stage it loses to $80-90{\mu}$. 6. Change in moisture content After pollination moisture content becomes gradually increased at the top in the early June and becomes markedly decreased in the middle of August. At the adult stage it shows 43~48% in cone, 23~25% in the outer seed-coat, 32~37% in the inner seed-coat, 23~26% in the inner seed-coat and endosperm and embryo, 21~24% in the embryo and endosperm, 36~40% in the embryos. 7. The content compositions of seed 1) Fat contents become gradually increased after the early May, at the adult stage it occupies 65~85% more fat than walnut and palm. Embryo includes 78.8% fat, and 57.0% fat in endosperm. 2) Sugar content after pollination becomes greatly increased as in the case of reducing sugar, while non-reducing sugar becomes increased in the early June. 3) Crude protein content becomes gradually increased after the early May, and at the adult stage it becomes 48.8%. Endosperm is made up with more protein than embryo. 8. The test of germination The collected optimum period of Pinus koraiensis seeds at an adequate maturity was collected in the early September, and used for the germination test of reduction-method and embryo culture. Seeds were taken at the interval of 7 days from the middle of July to the middle of September for the germination test at germination apparatus.