Abdul Murad, Nor Azian;Razak, Zuraini Abdul;Hussain, Rosniza Muhammmad;Syed Hussain, Sharifah Noor Akmal;Ching Huat, Clarence Ko;Siti Aishah, Che Md. Ali;Abdullah, Norlia;Muhammad, Rohaizak;Ibrahim, Naqiyah;Jamal, Rahman
Asian Pacific Journal of Cancer Prevention
/
v.14
no.3
/
pp.1655-1659
/
2013
Background: HER-2/neu is a proto-oncogene that encodes a transmembrane tyrosine kinase growth factor which is crucial for stimulating growth and cellular motility. Overexpression of HER-2/neu is observed in 10-35% of human breast cancers and is associated with pathogenesis, prognosis as well as response to therapy. Given the imperative role of HER-2/neu overexpression in breast cancer, it is important to determine the magnitude of amplification which may facilitate a better prognosis as well as personalized therapy in affected patients. In this study, we determined HER-2/neu protein expression by immunohistochemistry (IHC) concurrently with HER-2/neu DNA amplification by quantitative real time-polymerase chain reaction (Q-PCR). Materials and Methods: A total of 53 paired tissue samples from breast cancer patients were frozen-sectioned to characterize the tumour and normal tissues. Only tissues with 80% tumour cells were used in this study. For confirmation, Q-PCR was used to determine the HER-2/neu DNA amplification. Results: We found 20/53 (37.7%) of the tumour tissues to be positive for HER-2/neu protein overexpression using IHC. Out of these twenty, only 9/53 (17%) cases were in agreement with the Q-PCR results. The concordance rate between IHC and Q-PCR was 79.3%. Approximately 20.7% of positive IHC cases showed no HER-2/neu gene amplification using Q-PCR. Conclusion: In conclusion, IHC can be used as an initial screening method for detection of the HER-2/neu protein overexpression. Techniques such as Q-PCR should be employed to verify the IHC results for uncertain cases as well as determination of HER-2/neu gene amplification.
Kim, Tae-Rim;Yoon, Seog-Young;Heo, Jin-Young;Lee, Chi-Seung
Journal of the Korean Crystal Growth and Crystal Technology
/
v.27
no.5
/
pp.235-242
/
2017
In order to prepare porous scaffolds capable of pore control, PMMA powder serving as a pore-forming agent was added to HA powder to synthesize a slurry containing TBA as a solvent. And then, porous HA scaffolds where pillarshaped pore channels interconnected with each other were fabricated by freeze-casting and sintering. The crystal structure of the HA scaffolds according to the addition amount of PMMA powder was measured by XRD and the surface and inner cross section of the scaffolds were analyzed through SEM. It was found that removal of PMMA during sintering affects the internal structure of the scaffolds and the crystallinity of the HA powder. Furthermore, through evaluating the physical and mechanical properties of the scaffolds, it was confirmed that the porosity, pore size and compressive strength can be controlled by controlling the addition amount of the pore-forming agent. It was also found that the HA scaffolds produced in this study were similar in structure and properties to the natural cancellous bone. This suggests that porous HA scaffolds with PMMA can be used as an alternative to autogenous bone for tissue engineering as an artificial bone scaffold.
Purpose: The modified minimally invasive surgical technique (M-MIST) has been successfully employed to achieve periodontal regeneration. Platelet-rich fibrin (PRF) is known to enhance wound healing through the release of growth factors. This study aimed to observe the outcomes of periodontal surgery when M-MIST was used with or without PRF for the treatment of isolated intrabony defects. Methods: This randomized clinical trial was conducted on 36 systemically healthy patients, who had chronic periodontitis associated with a single-site buccal probing pocket depth (PPD) and clinical attachment level of ≥5 mm. Patients were randomly divided into 2 groups: the test group treated with M-MIST and PRF, and the control group treated with M-MIST alone. The primary periodontal parameters analyzed were PPD, relative attachment level (RAL), and relative gingival margin level. The radiographic parameters analyzed were change in alveolar crest position (C-ACP), linear bone growth (LBG), and percentage bone fill (%BF). Patients were followed up to 6 months post-surgery. Results: Intragroup comparisons at 3 and 6 months showed consistently significant improvements in PPD and RAL in both the groups. In intergroup comparisons, the improvement in PPD reduction, gain in RAL, and the level of the gingival margin was similar in both groups at 3 and 6 months of follow-up. Furthermore, an intergroup comparison of radiographic parameters also demonstrated similar improvements in C-ACP, LBG, and %BF at 6 months of follow-up. Conclusions: M-MIST with or without PRF yielded comparable periodontal tissue healing in terms of improvements in periodontal and radiographic parameters. Further investigation is required to confirm the beneficial effects of PRF with M-MIST.
Kim, Eun-Jung;Park, Yong-Won;Kim, Young-Han;Kim, Yu-Seun;Oh, Jung-Tak
Advances in pediatric surgery
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v.15
no.1
/
pp.1-10
/
2009
Recently, amniotic fluid has gained attention as one of the potential sources for cell therapy and tissue engineering because it has characteristics of multipotent stem cells. However, current knowledge about what types of cells are naturally found in amniotic fluid is still limited. In this study, we aimed to investigate whether human amniotic fluid contains cells that have characteristics of respiratory cells. Samples of human amniotic fluid (5 mL per sample) obtained from amniocenteses were cultured with small airway growth medium (SAGM). Cells were grown until the third passage and the presence of type II alveolar cells were characterized by inverted microscopy, immunofluorescence, and reverse transcription polymerase chain reaction (RT-PCR). On inverted microscopy, cultured cells showed typical polygonal and cobblestone-like epithelial morphology. The morphology of cells was not changed after selection and passing. Immunofluorescence analysis demonstrated that the isolated cells stained positive for surfactant protein C (SPC), specific marker for type II alveolar cells. Cells also stained positive for TTF-1 protein but negative for CD 31 and vimentin. RT-PCR analysis of cells showed expression of SPC mRNA. This study has demonstrated that respiratory cells can be isolated and identified from human amniotic fluid cultured in SAGM medium. Our results may provide the basis for further investigations of amniotic fluid.
This study was carried out with Lilium oriental hybrid cv. Casa Blanca to observe the effect of in vitro culture conditions on ex vitro sprouting of bulblets. Low temperature (15$^{\circ}C$) inhibited the growth of in vitro bulblets while high temperature ($25^{\circ}C$) enhanced the growth. Bulblets cultured at 15$^{\circ}C$ did not show dormacy while those cultured at 2$0^{\circ}C$ ,$25^{\circ}C$ had a longer dormancy period. High sucrose concentration (9%) induced longer dormancy. Dormancy period was also prolonged in bulblets cultured in vitro at high temperature ($25^{\circ}C$). Dormancy period was more affected by in vitro culture temperature rather than sucrose concentration. Physiological dormancy was released more rapidly when bulblets were cultured at $25^{\circ}C$ for 6 weeks and further transferred at 15$^{\circ}C$ and cultured for another 12 weeks. Treatment of ABA induced the dormancy in Lilium bulblets but when bulblets were subjected to chilling treatment (4$^{\circ}C$ for 8 weeks) nearly 100% sprouting were observed. The medium containing 1.0 mg/L BA or 1.0 mg/L fluridone was also effective to produce non-dormant bulblets.
This study was carried out to find the effects of plant growth regulators on callus formation, rooting and shooting from cotyledon explant in oriental me]on. Various combinations of 0.1 mg/L auxins (IAA, NAA) and 0.5, 1.0. 1.5, 2.0 mg/L cytokinins (BA, kinetin, zeatin) were treated to the MS basal medium, respectively. Callus was induced mort effectively as 2,437.0 mg (FW)/explant in MS medium supplemented with 0.1 mg/L NAA and 2.0 mg/L BA, but that was non-embryogenic callus as colored yellow white and broke easily. Root was induced most effectively at a frequency of 98.0% in MS medium supplemented with 0.1 mg/L NAA and 0-5 mg/L kinetin. Shoots formed on cut part of vein at a frequency of 98.0% in MS medium supplemented with 0.1 mg/L IAA and 2.0 mg/L BA, that were multiple shoots. in case of its concentration, BA and lower concentration of IAA and NAA (0.01 and 0.05 mg/L). respectively. shooting ratio was not increased. The result of treatment with BA 0-5 mg/L and IAA 0.1 mg/L, callus induced at a week, and shoot start to form multiple shoots about 3 weeks after inoculation. After 2 times subculture as 2 weeks intervals, divided shoots rooted and developed into intact plantlets at 10 weeks and then that grown normally on pots after acclimatization.
Kim, Kyung-Hee;Kim, Yong-Goo;Heo, Sung-Hyun;Bae, Eun-Ji;Lee, Kwang-Soo;Park, Nam-Chang;Lee, Byung-Hyun
Asian Journal of Turfgrass Science
/
v.25
no.2
/
pp.184-189
/
2011
To optimize tissue culture conditions for genetic transformation of Zoysia matrella (L.) Merr., we investigated the effects of different plant growth regulators and medium supplements on plant regeneration using stolon explants excised from mature plant grown in a green house. Plant regeneration frequency was 33.3% when stolon tissues with a node were cultured on the regeneration medium supplemented with 0.5 mg/L 2,4-D and 1 mg/L of kinetin. Comparing the basal media tested, MS medium showed higher plant regeneration performance than N6 or SH medium. Addition of 5 mg/L $AgNO_3$ with 10 mg/L cysteine improved frequency of plant regeneration up to 40%. Among different carbon sources, 3% sucrose was found to show the best for regeneration frequency. This rapid and efficient plant regeneration system would be useful for using genetic transformation experiments of manilagrass without intervening callus-mediated regeneration.
Experiments were conducted to evaluate the nutritive values of supplemental L-lysine, liquid and powder type, and DL-methionine in weanling pigs. For feeding trial, 165 weanling pigs were treated in 2 controls; 18 and 16% CP, 6 supplementations of lysine alone to 16% CP diets; 0.1, 0.2 and 0.4% of liquid and powder type each, and 3 supplementations of lysine + methionine to 15% CP diets; 0.05 + 0.025, 0.1 + 0.05 and 0.2 + 0.1%. Pigs were fed for 5 week to investigate the protein sparing effect of supplemental amino acid, and the optimal supplemental level. A metabolic trial included the measurements of digestibilities of dry matter, crude protein, crude fat, crude fiber, energy, phosphorus and amino acids. The liver acinar cell culture was conducted for the protein synthesis activity of the pigs fed each experimental diet. Supplementation of both type of L-lysine in 16% CP diet showed improved daily weight gain and feed efficiency which were compatible with those of pigs fed 18% CP diet. Groups fed liquid lysine did not differ from those fed powder type in growth performance. Supplementation of lysine and methionine to 15% CP diet did not improve growth performance of pigs to the extent that 18% CP diet was fed. In nutrient digestibility, 16% CP control diet showed significantly (p < 0.05) lower crude protein digestibility than any other treatments. Digestibilities of 16% CP diets with lysine supplementation were equal to that of 18% CP control, while digestibilities of 15% CP diets with the supplementation of lysine + methionine was inferior to that of 18% CP control. Supplementation of lysine alone reduced the nitrogen excretion compared to the none supplemented control groups. However, addition of lysine + methionine excreted more nitrogen than controls. Pigs fed diet supplemented with lysine alone, or lysine + methionine excreted less fecal phosphorus than those fed none supplemetation. Retained protein from liver tissue of pigs fed 18% diet was significantly (p < 0.05) greater than those fed 16% CP diet. A significant difference (p < 0.05) was observed in physical type of lysine. Feeding of powder type showed less secreted protein and greater retained protein in the culture of liver acinar cell. It is concluded that supplementation of lysine at the level of 0.1 to 0.2% can spare 2% of dietary protein and reduce nitrogen excretion by 19.3%. Also, no difference in nutritional values was observed between liquid and powder lysine in weanling pigs.
The effect of heat or chemotherapeutant treatment on the mosaic virus infected garlic(Allium sativum L.) scales and that of chemotherapeutant added to the culture medium were summarized as following. The treatment of the virus infected garlic scales at $37\~57^{\circ}C$ for 35 days to one hour in water or in air shelved no effort of inactivating the virus. Although treatment of the garlic scales at $62\~72^{\circ}C$ for 90 to five minutes reduced the mosaic symptom on the leaves of the garlic plants grown after the heat treatment, it reduced the growth vigor of the plants so greatly that complete inactivation of the virus in garlics was not feasible. The mosaic symptom on the leaves of garlic plant was reduced when the infected garlic scales were grown after 24 hours soaking in $10\~50\;ppm$ Malachite Green, 2,4-Dichlorophnoxy Acetic Acid, or in $20\~100\;ppm$ Quinhydron. These chemotherapeutants, however, inhibited the growth of garlic plant at the high concentration. Garlic scales soaked in $10\~50\;ppm$ Naphthyl Acetic Acid showed the least mosaic symptom without its complete extiction on the garlic leaves. When incorporated into the modified Murashige-Skoog's medium, $0.5\~l.5\;ppm$ Naphthyl Acetic Acid could inactivate the mosaic virus in newy developed garlic plants showing no mosaic symptom on the leaves, no inclusion bodies and intact nuclei in the leaf tissue cells.
Proceedings of the Plant Resources Society of Korea Conference
/
2010.05a
/
pp.17-17
/
2010
Ethylene, known as a stress hormone regulate wide developmental processes including germination, root hair initiation, root and shoot primordial formation and elongation, leaf and flower senescence and abscission, fruit ripening. The acceleration of ethylene biosynthesis in plant associated with environmental and biological stresses. 1-Aminocycloprophane-1-carboxlyate deaminase(ACCD) is an enzyme that cleaves ACC into and ammonia, a precursor of the plant hormone ethylene. Plant growth-promoting rhizobacteria (PGPR) having ACCD can decrease endogenous ACC level of tissue, resulting in reduced production of ethylene in plants. ACC deaminse was a key enzyme for protect stressed plants from injurious effects of ethylene. ACCD gene was encoded from Pseudomonas flourescens, PGPR and was cloned in Escherichia coli. We expressed the recombinant ACCD(rACCD) containing 357 amino acids with molecular weight 39 kDa that revealed by SDS-PAGE and western blot. The rACCD was purified by Ni-NTA purification system. The active form of rACCD having enzyme activity converted ACC to a-ketobutyrate. The optimal pH for ACC deaminase activity was pH 8.5, but no activity below pH 7.0 and a less severe tapering activity at base condition resulting in loss of activity at over pH 11. The optimal temperature of the enzyme was $30^{\circ}$ and a slightly less severe tapering activity at 15 - 30$^{\circ}$, but no activity over $35^{\circ}$. P. flourescens ACC deaminase has a highly conserved residue that plays in allowing substrate accessibility to the active sites. The enzymatic properties of this rACCD will provide an important reference for analysis of newly isolated ACCD and identification of newly isolated PGPR containing ACCD.
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