Ralstonia eutropha NCIMB 11599 and ATCC 17699 were grown, and their productions of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] compared. In flask cultures of R. eutropha NCIMB 11599, cell concentration, P(3HB-co-4HB) concentration and polymer content decreased considerably with increases in the ${\gamma}-butyrolactone$ concentration, and the 4HB fraction was also very low (maximum 1.74 mol%). In fed-batch cultures of R. eutropha NCIMB 11599, glucose and ${\gamma}-butyrolactone$ were fed as the carbon sources, under a phosphate limitation strategy. When glucose was fed as the sole carbon source, with its concentration controlled using an on-line glucose analyzer, 86% of the P(3HB) homopolymer was obtained from 201g/L of cells. In a two-stage fed-batch culture, where the cell concentration was increased to 104g/L, with glucose fed in the first step and constant feeding of ${\gamma}-butyrolactone$, at 6g/h, in the second, final cell concentration at 67h was 106g/L, with a polymer content of 82%, while the 4HB fraction was only 0.7mol%. When the same feeding strategy was applied to the fedbatch culture of R. eutropha ATCC 17699, where the cell concentration was increased to 42 g/L, by feeding fructose in the first step and ${\gamma}-butyrolactone$ (1.5g/h) in the second, the final cell concentration, polymer content and 4HB fraction at 74h were 51g/L, 35% and 32 mol%, respectively. In summary, R. eutropha ATCC 17699 was better than R. eutropha NCIMB 11599 in terms of P(3HB-co-4HB) production with various 4HB fractions.
In this study, we investigated the anti-proliferative effect of polysaccharides from Salicornia herbacea on HT-29 human colon cancer cells. Crude polysaccharides from S. herbacea (CS) were prepared by extraction with hot steam water, and fine polysaccharides from S. herbacea (PS) were obtained through further size exclusion chromatography. The anti-proliferative effect of CS and PS were measured using the MTS assay, apoptosis analysis, cell cycle analysis, and RT-PCR. HT-29 cells were treated with CS or PS at different dosages (0.5, 1, 2, 4 mg $ml^{-1}$) for 24 or 48 h. CS and PS inhibited proliferation and stimulated apoptosis of cells in a dose-dependent manner. Flow cytometric analysis after Annexin V-FITC and PI staining revealed that treatment with CS or PS increased total apoptotic death of cells to 24.99% or 91.59%, respectively, in comparison with the control (13.51 %). PS increased early apoptotic death substantially - up to 12 times more than the control. Treatment with CS or PS resulted in a concentration-dependent increase of the G2/M cell population of the cell cycle as determined by flow cytometry. G2/M arrest was induced significantly with the highest concentration (4 mg $ml^{-1}$) of PS. RT-PCR was performed to study the correlation between G2/M arrest and transcription of cell cycle control genes. The anti-proliferative activity of CS and PS was accompanied by inhibition of cyclin B1, and Cdc 2 mRNA. Moreover, both CS and PS induced expression of the p53 tumor suppressor gene and the Cdk inhibitor p21. These results suggest that polysaccharides from S. herbacea have anti-cancer activity in human colon cancer cells.
Park, Sang Ah;Jeong, Hae Jin;Ok, Jin Hee;Kang, Hee Chang;You, Ji Hyun;Eom, Se Hee;Yoo, Yeong Du;Lee, Moo Joon
ALGAE
/
v.36
no.4
/
pp.299-314
/
2021
Some species in the dinoflagellate genus Alexandrium are bioluminescent. Of the 33 formally described Alexandrium species, the bioluminescence capability of only nine species have been tested, and eight have been reported to be bioluminescent. The present study investigated the bioluminescence capability of seven Alexandrium species that had not been tested. Alexandrium mediterraneum, A. pohangense, and A. tamutum were bioluminescent, but A. andersonii, A. hiranoi, A. insuetum, and A. pseudogonyaulax were not. We also measured the bioluminescent intensity of A. affine, A. fraterculus, A. mediterraneum, A. ostenfeldii, A. pacificum, A. pohangense, A. tamarense, and A. tamutum. The mean 200-second-integrated bioluminescence intensity per cell ranged from 0.02 to 32.2 × 104 relative luminescence unit per cell (RLU cell-1), and the mean maximum bioluminescence intensity per cell per second (BLMax) ranged from 0.01 to 10.3 × 104 RLU cell-1 s-1. BLMax was significantly correlated with the maximum growth rates of Alexandrium species, except for A. tamarense. A phylogenetic tree based on large subunit ribosomal DNA (LSU rDNA) showed that the bioluminescent species A. affine, A. catenella, A. fraterculus, A. mediterraneum, A. pacificum, and A. tamarense formed a large clade. However, the toxicity or mixotrophic capability of these species was split. Thus, their bioluminescence capability in this clade was more consistent than their toxicity or mixotrophic capability. Phylogenetic trees based on LSU rDNA and the luciferase gene of Alexandrium were consistent except for A. pohangense. The results of the present study can provide a basis for understanding the interspecific diversity in bioluminescence of Alexandrium.
In this study, to assess the diagnostic value for mastitis in dairy cows, change of acute phase proteins(haptoglobin and serum amyloid A) concentrations in milk and sera of dairy cows were measured. 50 dairy cows were used in this experiment and divided into two groups. The first group was the healthy dairy cow group whose milk contained less than 2.0${\times}$10$\^$5/ somatic cell counts(n=5). The second group was the mastitis-dairy cow group whose milk counted higher than 5.0${\times}$10$\^$5/ somatic cell counts(n=45). The concentration of haptoglobin and serum amyloid A in milk and sera from these two groups were determined by Tridelta range haptoglobin kit and serum amyloid A kit. The concentration of haptoglobin in the milk from first group was undetectable value and that of the second group was 124.0$\mu\textrm{g}$/$m\ell$. And the concentration of haptoglobin in serum of the first group was 32.0$\mu\textrm{g}$/$m\ell$ and that of the second group was 214.4$\mu\textrm{g}$/$m\ell$. The concentration of serum amyloid A in the milk from first group was 0.32$\mu\textrm{g}$/$m\ell$ and that of the second group was 17.7$\mu\textrm{g}$/$m\ell$. And the concentration of serum amyloid A in serum of the first group was 5.1$\mu\textrm{g}$/$m\ell$ and that of the second group was 25.8$\mu\textrm{g}$/$m\ell$. It was concluded that concentration of haptoglobin and serum amyoid A in milk and serum may be was to discriminate between normal and mastitic milks.
In cervical cancer, one of the most common malignant tumors in women worldwide, miR-126 has been reported to exhibit decreased expression. However, its role in cervical cancer cell proliferation and drug sensitivity has remained relatively unexplored. Here, we compared the expression of miR-126 in cervical cancer tissues (n = 20) with that in normal cervical tissue (n = 20) using quantitative RT-PCR. The viability of Siha cervical cancer cells was further measured by MTT assay after transfection with miR-126 mimic (Siha-miR-126 mimic) or microRNA mimic negative control (Siha-miR mimic NC) and after treatment with various concentrations of bleomycin (BLM). IC50s were calculated, and the survival rates (SRs) of Siha cells were calculated. miR-126 expression in cervical cancer tissue was significantly decreased compared with that in normal cervical tissue (P < 0.01). The relative SRs of Siha-miR-126 mimic cells were also significantly decreased compared with those of Siha-miR mimic NC cells at 24-96 h after transfection. The IC50 of BLM in Siha-miR-126 mimic cells ($50.3{\pm}2.02{\mu}g/mL$) was decreased compared with that in Siha-miR mimic NC cells ($70.5{\pm}4.33{\mu}g/mL$) at 48 h after transfection (P < 0.05). Finally, the SRs of Siha-miR-126 mimic cells were significantly lower than those of SihamiR mimic NC cells after cultured in medium containing 40 ${\mu}g/mL$ BLM for 24-96 h (P < 0.05). These results suggest that miR-126 is expressed at low levels in cervical cancer. Upregulation of miR-126 inhibited cervical cancer cell proliferation and enhanced the sensitivity to BLM. Thus, miR-126 may represent a novel approach to cervical cancer treatment.
Background: Cancer survivors are at increased risk of second cancers. Lymphoproliferative disorders (LPD) are common neoplasms that are primary or subsequent cancers in cases of multiple primary cancer. We here analyzed metachronous or synchronous LPD in multiple primary cancers. Methods: Between 2001 and 2010, LPD were assessed retrospectively in 242 multiple primary cancers patients. Results: Forty nine (20.2%) patients with LPD were detected. Six patients had two LPD where one patient had three LPD. The median age of patients was 60.5 years (range: 28-81). LPD were diagnosed in 29 patients as primary cancer, in 23 patients as second cancer, and in three patients as third cancer in multiple primary cancers. Primary tumor median age was 56 (range: 20-79). Diffuse large B cell lymphoma (n=16), breast cancer (n=9), and lung cancer (n=6) were detected as subsequent cancers. Alklylating agents were used in 19 patients (43.2%) and 20 patients (45.5%) had received radiotherapy for primary cancer treatment. The median follow-up was 70 months (range: 7-284). Second malignancies were detected after a median of 51 months (range: 7-278), and third malignancies with a median of 18 months (range: 6-72). Conclusions: In this study, although breast and lung cancer were the most frequent detected solid cancers in LPD survivors, diffuse large B cell lymphoma was the most frequent detected LPD in multiple primary cancers.
This study was conducted to optimize the efficiency of cloning and to produce cloned mice. The majority of cloned mammals derived by nuclear transfer (NT) die during gestation and have enlarged and dysfunctional placentas. In this study, the optimized conditions were established to produce clone mice. The parthenogenetic oocytes were activated after 6 h regardless of cytochalasin B (CB) concentration. CB treatment ($2{\mu}g/ml$) was found second polar body. Lower concentration of CB was decreased the activation rate, but the second polar body was the best highly increased during 6 h incubation. The small fragments were exhibited in the $5{\mu}g/ml$ treatment of CB, but it was not found in lower concentration groups (> $2.5{\mu}g/ml$). To examine effects of $SrCl_2$ on the adult cumulus cells, somatic cell NT oocytes were exposed during 0.5, 1 and 6 hrs. The second polar body was significantly greater in 0.5 h exposure group (6.6%) than 1, 6 hrs. Developmental rate from 2-cell to 4-cell was the lowest in 7.5 mM Strontium chloride ($SrCl_2$) groups (84.1% and 64.3%) than 5, 10 m $MSrCl_2$. The implantation rate was not significantly difference among 5, 7.5 and 10 m $MSrCl_2$ group. Three live fetuses were produced by SCNT. SCNT placentas were remarkably heavier than IVF group (8 fetuses) (0.34, 0.34, 0.33 vs 0.14 g) compared with the placenta weight of IVF and SCNT clones.
Background: The aim of this study was to investigate the effect of a Lipofectamine2000 (Life2000) Transfection Reagent transfected psiRNA-STAT3 plasmid on 4T1 breast cancer cells. Materials and Methods: MTT was used to detect the cell proliferation of breast cancer 4T1 cells at different periods (0h, 6h, 8h, 10h); the cell cycle was assessed by flow cytometry; variation of apoptosis and mitochondrial membrane potential was observed under a fluorescence microscope; immunohistochemical staining was used to determine the expression of caspase-3 and cyclin-D1 protein. Results: An obvious effect of inhibition to 4T1 cancer cells could be observed at 8h after the psiRNA-STAT3 was transfected. Typical alterations of apoptotic morphological features were visible in the psiRNA-STAT3 treatment group. Mitochondrial membrane potential decreased significantly, the number of cells was increased in G0/G1 phase, and the number of cells was decreased in S phase, and the data were statistically significant (p<0.05), compared with the Scramble and Mock groups. Expression of caspase-3 protein was increased significantly, while that of cyclin D1 was significantly decreased. Conclusions: Life2000 transfected psiRNA-STAT3 plasmid can inhibit 4T1 tumor cell proliferation and promote apoptosis of 4T1 tumor cells, which process depends on the regulation of expression of cyclin D1 and caspase-3 protein.
The Korea Electric Power Research Institute (KEPRI) has been developing planar solid oxide fuel cells (SOFCs) and power systems for combined heat and power (CHP) units. The R&D work includes solid oxide fuel cell (SOFC) materials investigation, design and fabrication of single cells and stacks, and kW class SOFC CHP system development. Anode supported cells composed of Ni-YSZ/FL/YSZ/LSCF were enlarged up to $15{\times}15\;cm^2$ and stacks were manufactured using $10{\times}10\;cm^2$ cells and metallic interconnects such as ferritic stainless steel. The first-generation system had a 37-cell stack and an autothermal reformer for use with city gas. The system showed maximum stack power of about $1.3\;kW_{e,DC}$ and was able to recover heat of $0.57{\sim}1.2\;kW_{th}$ depending on loaded current by making hot water. The second-generation system was composed of an improved 48-cell stack and a prereformer (or steam reformer). The thermal management subsystem design including heat exchangers and insulators was also improved. The second-generation system was successfully operated without any external heat source. Under self-sustainable operation conditions, the stack power was about $1.3\;kW_{e,DC}$ with hydrogen and $1.2\;kW_{e,DC}$ with city. The system also recuperated heat of about $1.1\;kW_{th}$ by making hot water. Recently KEPRI manufactured a 2kW class SOFC stack and a system by scaling up the second-generation 1kW system and will develop a 5kW class CHP system by 2010.
This Investigation was carried out to map the morphological development of serotonin-immunoreactive neurons in the larval brain of the cabbage butterfly, Artogeia rapae, during five larval stages. Both the first instar larva and the second instar larva contained twenty serotonin-immunoreactive (5-HTi) neurons in each brain. The fibres of 5-HTI commissure was interconnected to two cerebral hemispheres in both brains. However, the 5-HTi commissural fibres was Increased in number in the second-instar larva brain. In the brain of the second Insar larva these 5-HTi fibres formed rich arborization in contralateral neuropils, especially In the posterior parts of it. The third-Instar larva braIn, which Included twenty two 5-HTi neurons, had three groups of 5-HTi commissural fibres. In the fourth Instar larva, the number of 5-HTi fibres as well as 5-HTi cell bodies increased in the brain. The fifth-instar larva brain, which contained fifty four 5-HTi cell bodies, showed the largest number of 5-HTi cell bodies In developing larval brains. The 5-HTi fibres formed richest commissural fibres and some of them run parallel to anteroposterior axis.
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