Lee Jai-Heon;Kim Kee-Young;Chung Dae-Soo;Chung Won Bok;Kwon Oh-Chang
Proceedings of the Korean Society of Crop Science Conference
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1999.05a
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pp.162-163
/
1999
The goal of this research was (1) to describe the conditions and parameters required for the cell cycle synchronization and the accumulation of large number of metaphase cells in maize and other cereal root tips, (2) to isolate intact metaphase chromosomes from root tips suitable for characterization by flow cytometry, and (3) to construct chromosome-specific libraries from maize. Plant metaphase chromosomes have been successfully synchronized and isolated from many cereal root-tips. DNA synthesis inhibitor (hydroxyurea) was used to synchronize cell cycle, follwed by treatement with trifluralin to accumulate metaphase chromosomes. Maize flow karyotypes show substantial variation among inbred lines. thish variation should be sueful in isolating individual chromosome types. In addition, flow cytometry is a useful method to measure DNA content of individual chromosomes in a genotyps, and to detect chromosomal variations. Individual chromosome peaks have been sorted from the maize hybrid B73/Mol7. Libraries were generated form the DOP-PCR amplification product from each peak. To date, we have analyzed clones from a library constructed from the maize chromosome 1 peak. Hybridization of labeled genomic DNA to clone inserts indicated that $24\%,\;18\%,\;and\;58\%$ of the clones were highly repetitive, medium repetitive, and low copy, respectively. Fifty percent of putative low cpoy clones showed single bands on inbred screening, blots, and the remaining $50\%$ were low copy repeats. Single copy clones showing polymorphism will be mapped using recombinant inbred mapping populations. Repetitive clones are being characterized by Southern blot analysis, and will be screened by in situ hybridization for their potential utility as chromosome specific markers.
The spinal dorsal horn is the area where primary afferent fibers terminate and cutaneous sensory information is processed. A number of putative neurotransmitter substances, including excitatory and inhibitory amino acids and peptides, are present in this region. In this study, single neurons of the spinal dorsal horn were acutely isolated and the properties of whole cell current and responses to excitatory and inhibitory neurotransmitters were studied by patch clamp technique. Transverse slice ($(300{\mu}m$) of lumbar spinal cords from young rats$(7{\sim}14\;days)$ were sequentially treated with two pretenses(pronase 0.75 mg/ml and thermolysin 0.75 mg/ml), then single neurons were mechanically dissociated. These neurons showed near-intact morphology such as multipolar, ellipsoidal and bipolar, and pyramidal cells and we recorded the typical whole cell currents of $K^+$, $Ca^{2+}$ and ligand-operated channels from these neurons. Glutamate $(30{\mu}M)$ and N-methyl-D-aspartate(NMDA, $30{\mu}M)$ induced inward currents of $117{\pm}12.4$ pA(n=5) and $49{\pm}6.9$ pA(n=3), respectively. Glycine $(1{\mu}M)$ potentiated glutamate-induced currents $4{\sim}5$ times and NMDA-induced currents $8{\sim}10$ times. In addition, glycine $(30{\mu}M)$ induced Inward current ($31{\pm}6.1$ nA, n=2), which was rapidly desensitized after the peak to a new steady-state level. However, the inward currents induced by ${\gamma}-amino$ butyric acid(GABA, $1{\mu}M$) decreased continuously after the peak($226{\pm}41.6$ pA, n=3) under the similar experimental condition. The ionic currents and pharmacological responses of isolated neurons in this work were similar to those observed in vivo or in vitro spinal cord slice, indicating that acutely isolated neurons could be effectively used for further pharmacological studies.
Sex hormones have long been considered to play an important role in bone turnover rate, periodontal diseases, and wound healing. We have studied the effect of porcine testis steroid extract (PTSE), an extract of porcine testes, which holds a good ratio of 19-nortestosterone (nandrolone), testosterone, androstenedione, $17{\beta}$-estradiol, and estrone, on the healing rate of a standardized full-thickness linear wound on the back of the rat. Skin punch or carbon dioxide ($CO_2$) laser methods were used to create the deep skin injury in two groups of animals. The animals were treated with the PTSE cream, control cream and Vaseline (control) to find out the effect in re-epithelialization, contraction, and formation of granulation and scar tissues. Histological examination after 21 days showed 100, 87.4, and 80.5% recovery of epidermis, dermis, and hypodermis, respectively in the PTSE-treated animals. Similarly, on the 15th day of treatment, complete healing of intact skin was observed in the PTSE cream-treated animals among the laser radiation group. Even though the beginning of re-epithelialization phase and completion of serum crust formation was also observed in the base cream- and Vaseline-treated animals respectively, the complete healing cycle was observed only in the PTSE-treated group. The white blood cell count in the PTSE-treated group showed that PTSE cream is nontoxic to animals.
Kim, Sam Woong;Jeong, In Sil;Jeong, Eun Ju;Tak, Je Il;Lee, John Hwa;Eo, Seong Kug;Kang, Ho Young;Bahk, Jeong Dong
Molecules and Cells
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v.26
no.1
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pp.26-33
/
2008
The plasmid pJB01, a member of the pMV158 family isolated from Enterococcus faecium JC1, contains three open reading frames, copA, repB, and repC. Plasmids included in this family produce counter-transcribed RNA (ctRNA) that contributes to copy number control. The pJB01 ctRNA, a transcript which consists of 54 nucleotides (nts), is encoded on the opposite strand from the copA/repB intergenic region and partially overlaps an atypical ribosome binding site (ARBS) for repB. The ARBS is integrated by the two underlined conserved regions: 5'-TTTTTGTNNNNTAANNNNNNNNNATG-3', and the ctRNA is complementary only to the 5' conserved sequence 5'-TTTTTGT-3'. This complementary sequence is located at a distance from the terminal loop of the ctRNA secondary structure. The ctRNA structure predicted by the mfold program suggests the possible generation of a terminal and an internal hairpin loop. The amount of in vitro translation product of repB mRNA was inversely proportional to the ctRNA concentration. Mutations in the terminal and internal hairpin loops of the ctRNA had inhibitory effects on its binding to the target mRNA. We propose that the intact structures of the terminal and internal hairpin loops, respectively, play important roles in forming the initial kissing and extending complexes between the ctRNA and target mRNA and that these regulate the copy number of this plasmid.
Objective : This study was carried out to understand effects of ginseng(hearinafter ; GS, Panax Ginseng) extract on regeneration responses on injured sciatic nerves in rats. Methods :Using white mouse, we damaged sciatic nerve & central nerve, and then applied GS to the lesion. Then we observed regeneration of axon and non-neuron. Results : 1. NF-200 protein immunostaining for the visualization of axons showed more distal elongation of sciatic nerve axons in GS-treated group than saline-treated control 3 and 7 days after crush injury. 2. GAP-43 protein was increased in the injured sciatic nerve and further increased by GS treatment. Enhanced GAP-43 protein signals were also observed in DRG prepared from the rats given nerve injury and GS treatment. 3. GS treatment in vivo induced enhanced neurite outgrowth in preconditioned DRG sensory neurons. In vitro treatment of GS on sensory neurons from intact DRG also caused increased neurite outgrowth. 4. Phospho-Erk1/2 protein levels were higher in the injured nerve treated with GS than saline. Phospho-Erk1/2 protein signals were mostly found in the axons in the injured nerve. 5. NGF and Cdc2 protein levels showed slight increases in the injured nerves of GS-treated group compared to saline-treated group. 6. The number of Schwann cell population was significantly increased by GS treatment in the injured sciatic nerve. GS treatment with cultured Schwann cells increased proliferation and Cdc2 protein signals. 7. GS pretreatment into the injured spinal cord generated increased astrocyte proliferation and oligodendrocytes in culture. In vitro treatment of GS resulted in more differentiated pericytoplasmic processes compared with saline treatment. 8. More arborization around the injury cavity and the occurrence at the caudal region of CST axons were observed in GS-treated group than in saline-treated group. Conclusion :GS extract may have the growth-promoting activity on regenerating axons in both peripheral and central nervous systems.
Objectives : The purpose of this study was to investigate the effects of warm needling at $GB_{30}{\cdot}GB_{34}$ on Complete Freund's Adjuvant(CFA)-induced rheumatoid arthritis in rats. Methods : Arthritis was induced by injecting CFA subcutaneously into the left knee joint and paw. Acupuncture(AT) and warm needling(W-AT0.5, W-AT1.0) were injected at $GB_{30}{\cdot}GB_{34}$, every other day for a total of 5 times beginning on day 10 after the CFA injection. Thereafter, external shape, paw edema, serum aminotransferase and anti-inflammatory factors were assessed, and hematological and histological observations were made. Results : In paw edema volume all 3 groups(AT, W-AT0.5, W-AT1.0) showed significant decrease compared to the CFA control group. In TNF-${\alpha}$ and IL-6, all 3 groups showed significant decrease compared to CFA control group. In AST and ALT all 3 groups showed no significant change. In IL-$1{\beta}$, W-AT0.5 and W-AT1.0, groups showed significant decrease compared to the CFA control group. Leucocyte, erythrocyte and thrombocyte, all 3 groups showed no significant change. In histological observations, all 3 groups were similar to the intact group in terms of synoviocyte, cartilage lacuna and cartilage cells. Conclusions : The results suggest that warm needling at $GB_{30}{\cdot}GB_{34}$, has the effect of suppressing inflammation of CFA-induced rheumatoid arthritis in rats.
Objectives : This experiments were performed to determine the effect of OYakSoonGi-San extract on hypertension in spontaneous hypertensive rat and norepinephrine-induced arterial contraction in rabbit. Methods : In order to define the effect of OYakSoonGi-San extract on contracted rabbit carotid arterial strips, transverse strips with intact or damaged endothelium were used for the experiment using organ bath. To analyze the mechanism of OYakSoonGi-San extract-induced relaxation, OYakSoonGi-San extract infused into contracted arterial strips induced by norepinephrine after treatment of indomethacin, $N{\omega}-nitro-L-arginine$, methylene blue or tetraethylammonium chloride. Results : Blood pressure was significantly decreased five days after administration of OYakSoonGi-San extract. The relaxation effect of OYakSoonGi-San extract was dependent on the presence of endothelium, showing that OYakSoonGi-San extract-induced relaxation was not observed in the strips without endothelium. Also OYakSoonGi-San extract-induced relaxation was significantly inhibited in arterial strips which were contracted by high $K^+$. OYakSoonGi-San extract-indeced relaxation was significantly inhibited by the pre-treatment of $N{\omega}-nitro-L-arginine$ or methylene blue, but it was not observed in the strips pre-treated with indomethacin or tetraethylammonium chloride. When additive application of $Ca^{2+}$ in arterial strips which were pre-contracted by norepinephrine in a $Ca^{2+}$-free solution, arterial contraction was increased. But contractile response to $Ca^{2+}$ was attenuated by pre-treatment of OYakSoonGi-San extract. Conclusions : These results demonstrated that OYakSoonGi-San could be applied effectively to hypertension and may inhibit agonist-induced contraction through an decrease influx of extra-cellular $Ca^{2+}$ by the formation of nitric oxide in the vascular endothelial cells.
For microbial production of acetaminophen, a popular analgesic-antipyretic from aniline, we screened various fungi and bacteria. And we succeeded to some extents in acetaminophen production by successful protoplast fusion between S. lividans and S. globisporus and also between S. rimosus and S. aureofaciens. However, more fertile results might be brought via performing the cloning of acetanilide p-hydroxylation genes of Streptomyces in yeast. This study was initiated to determine whether the acetanilide p-hydroxylase of Streptomyces is cytochrome P-450 species or non-heme iron protein species. The p-hydroxylationactivity on acetanilide in S. aureofaciens ATCC 10762 was found to be unstable on exposing to the air. However, 100,000xg supernatant of the cell free extracts which were prepared in $N_2$ atmosphere showed the p-hydroxylation activity. Characteristic absorption peak of cytochrome P-450 after reduction with dithionite and addition of CO was not observed in the region of 450nm. Moreover, metyrapone and 2, 6-dichloroindophenol did not affect this enzyme activity, but sodium azide, sodium cyanide, cupric sulfate, cadmium chloride, ${\alpha}$, ${\alpha}'-dipyridyl$, and o-phenanthroline reduced p-hydroxylase activity considerably. S. fradiae NRRL 2702 was shown to have strong p-hydroxylation activity in intact cells. This activity disappeared in its cell free extracts. In its 100,000xg supernatant, however, characteristic absorption peak of cytochrome P-450 after reduction with dithionite and addition of CO was observed at 446nm. Thus, the results herein presented suggest that acetanilide p-hydroxylase of Streptomyces aureofaciens is not related to cytochrome P-450 and may include non-heme iron protein for its activity. However, it is not clear whether acetanilide p-hydroxylase in S. fradiae belongs to the same category of S. aureofaciens.
Thymoma Is an anterior mediastinal tumor, arising from the thymus, but occasionally has ectopic focus such as neck, trachea, thyroid, pulmonary hilum, lung parenchyme and pleura. Forty-two year old male patient was admitted due to progressive development of shortness of breath in 4 months. He had a history of exposure to asbestos for About 10 years duration In recent 15 years. Radiologically, multiple pleural masses were seen rom apex to diaphrAgm, with no evidence of anterior mediastinal mass,with fluid in right pleural cavity. Closed thoracotomy drainage with open biopsy were performed. Effusion cell block showed many T cell marker positive Lymphocytes & some epithelial cells compatible with thymoma, and the tissue also showed cortical type thymoma. Pleuropneumonectomy and thymectomy followed by 60 Gy radiation therapy were done and the patient is well 8 months postoperatively. The pleura is markedly thickened by the invasion of thymoma and the interstitial space of the lung tissue,but the normal appearance of thymus was present in remote area (Masaoka classification IVa). We report a case of ectopic invasive thymoma arising from the right pleura with intact thymus.
A 9-year-old, intact female Jindo dog weighing 23 kg was presented to the Veterinary Medical Teaching Hospital of Chungnam National University with a history of two months of ocular discharge. Initial ocular examination showed a 2 cm elongated mass arising from the ventral bulbar conjunctiva of the left eye. Ocular ultrasound, performed to rule out any orbital and intraocular involvement, was negative. Thoracic radiographs and abdominal ultrasonography revealed no abnormalities. Fine needle aspiration (FNA) of mass was performed. Aspiration cytology showed the presence of a round cell population mostly characterized. A moderate amount of eosinophils was present among round cells. The mass was excised and identified hitologically as a mast cell tumor. Six months after surgical treatment, thoracic radiographs and abdominal ultrasonography did not reveal any clinical signs of local recurrence or metastatic disease.
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