Periodontal ligament (PDL) cells have been known as multipotential cells, and as playing an important rolesin periodontal regeneration. The PDL cells are composed of heterogeneous cell populations which have the capacity to differentiate into either cementoblasts or osteoblasts, depending on needs and conditions. Therefore, PDL cells have the capacity to produce mineralized nodules in vitro in mineralization medium which include ascorbic acid, ${\beta}$-glycerophosphate and dexamethasone. In spite of these well-known osteoblast like properties of PDL cells, very little is known about the molecules involved in the formation of the mineralized nodules in the PDL cells. In the present study, we analysed gene-expression profiles during the mineralization process of cultured PDL cells by means of a cDNA microarray consisting of 3063 genes. Nodules of mineralized matrix were strongly stained with alizarin red S on the PDL cells cultured in the media with mineralization supplements. Among 3,063 genes analyzed, 35 were up-regulated more than two-fold at one or more time points in cells that developed matrix mineralization nodules, and 38 were down-regulated to less than half their normal level of expression. In accord with the morphological change we observed, several genes related to calcium-related or mineral metabolism were induced in PDL cells during osteogenesis, such as IGF-II and IGFBP-2. Proteogycan 1, fibulin-5, keratin 5, ,${\beta}$-actin, ${\alpha}$-smooth muscle actin and capping protein, and cytoskeleton and extracellular matrix proteins were up-regulated during mineralization. Several genes encoding proteins related to apoptosis weredifferentially expressed in PDL cells cultured in the medium containing mineralization supplements. Dkk-I and Nip3, which are apoptosis-inducing agents, were up-regulated, and Btf and TAXlBP1, which have an anti-apoptosis activity, were down-regulated during mineralization. Also periostin and S100 calciumbinding protein A4 were down-regulated during mineralization.
Kim, E.Y.;Uhm, S.J.;Kim, M.K.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
Clinical and Experimental Reproductive Medicine
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v.23
no.3
/
pp.319-326
/
1996
The objective of this study was to investigate correlation between the morphology by microscopic assessments of surplus blastocysts produced in human IVF program and their cell number obtained by differential labelling method. For these experiments, 76 surplus human blastocysts were obtained from 36 patients on day 5 after IVF, the embryos were classified to early (ErB), early expanding (EEB), middle expanding (MEB), expanded blastocyst (EdB) according to their blastocoel expansion and zona thickness. When the ovum size and zona thickness of the classified blastocysts were measured using micrometer, although the embryos were produced in the same culture condition, there were significant variances in ovum size ($148.8 217.6{\mu}m$) and zona thickness ($1.2-14.4{\mu}m$). Total blastomere cell number counted after hoechst staining was increased by two to three fold during the transition period from ErB ($39.1{\pm}3.6$) to EdB ($(89.6{\pm}3.3)$) stage on day 5 after IVF. ICM ($11.9{\pm}1.8-22.2{\pm}4.3$) and TE ($24.5{\pm}3.6-70.0{\pm}7.7$) cell numbers using differential labelling were also showed the increased pattern according to the developmental level. Especially, EdB which showed poor ICM morphologically also indicated the low ICM cell number after differential labelling. This demonstrated that there is good correlation between the morphological assessment and the cell number. The count of ICM and TE nuclei using differential labelling can be used as an important criterion, if it is accompanied with morphological assessments, in selecting the better embryos for improving the pregnancy rates in human blastocyst transfer program.
Cho, Byung-Je;Hong, Jun Young;Kim, Mijeong;Song, Yeong Ok
Journal of the Korean Society of Food Science and Nutrition
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v.43
no.9
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pp.1380-1387
/
2014
The purpose of this study was to develop a mouthwash product with solid fermented oriental medicinal herb (OMH). Solid fermentation of magnolia, liquorice, and cnidium by Phellinus linteus mycelium was carried out successfully when 30% water was added to the medium, whereas 10% brown rice powder was required as an extra nutrient for solid fermentation of mint besides water. The amount of total phenol compounds and DPPH radical scavenging activity of OMH increased significantly (P<0.05) upon solid fermentation. Anti-microbial activities of fermented OMH also increased and were approximately 100-fold greater than those of unfermented samples. Oral pathogens such as Staphylococcus epidermis, Streptococcus pyogenes, Candida albicans, or Streptococcus mutans were used for determination of anti-microbial effects of OMH. Formulation of the mouthwash was developed based on the results of the sensory evaluation. Among seven formulas, the best formula chosen by the sensory evaluation was as follows: mouthwash prepared with 0.075% ethanol extract of solid fermented OMH as a main ingredient, 83.64% hot water extract of mint and clove (100:15, v/v) as a mouthwash base component, and other miscellaneous ingredients, including sodium fluoride, menthol, and surfactants. Data from a consumer's preference test with 30 participants, overall acceptance, and willingness to buy the product developed in this study were all significantly higher for the tested mouthwash compared to mouthwash on the market manufactured with OMH but with a different formula. Duration of freshness of the mouthwash after usage as determined by Breath Checker was not significantly different between the two samples, although the duration of our product was slightly longer than that of the commercial product mentioned above.
Catheters were placed into the external jugular veins of immature female rats. On the following day (day 28 of age), the animals were injected subcutaneously with pregnant mare serm gonadotropin(PMSG): 4IU(control) or 20IU(superovulation). Each animal was sequentially bled at Ohr and 12hr and subsequently at 6hr intervals until sacrifice at 72hr after PMSG. The superovulatory dose of PMSG significantly(P<0.05) increased the ovulatory response by 4.0 fold above controls. On the other hand, superovulated oocytes displayed considerably different stages of meiotic maturation: prophase I (14.7%), anaphase I (36.2%), telophase I (10.3%), metaphase I/II (32.4%), while in control rats a majority of the oocytes examined(94.0%) consistently showed a metaphase II configuration. Serum luteinizing hormone(LH) levels were determined by RIA. Both groups exhibited a similar time relationship with two distinct peaks: an initial slight rise at 0-18hr and a second sharp rise at 54-60hr. However, there was a marked change in the magnitude of LH levels between the two groups. In superovulated animals, prior to the second peak, overall LH levels were significantly(P<0.001) higher than controls. In contrast, at the peak occurring at 60hr, LH concentrations were significantly(P<0.001) reduced by 54% below that of control. Additionally, a maximum increase of mean ${\Delta}LH$ between two peaks was much less in superovulated as compared to control rats. The initial prolonged elevation of serum LH before 54hr in superovulated rats was found to result from actual cross-reaction of the injected PMSG with LH antibody in the assay, while a precipitous second elevation between 54hr and 60hr resulted primarily from an endogenous LH surge. This study clearly defines time-course features of serum LH in PMSG-treated rats. The overall results indicate that, following superovulatory treatment with PMSG, the increased ovulatory response is primarily associated with PMSG-derived intrinsic gonadotropin, and that the recovery of immature or asynchronously mature oocytes at ovulation may reult from the circulatory alteration of LH activity characterized by an initial prolonged elevation of serum LH and its subsequent attenuation.
Recently it has been shown that central dopaminergic system regulates the renal function and that intracerebroventricularly (icv) administered dopamine (DA) produces antidiuresis and antinaturiuresis, resembling icv norepinephrine, and evidence has been accumulated which would suggest the involvement of adrenergic system in the DA effects. It was attempted therefore in this study to see whether the DA effect is influenced by pretreatment of yohimbine which is known as a specific ${\alpha}_2-adrenoceptor$ antagonist. Yohimbine produced, when given icv in doses of $100\;{\mu}g/kg$, marked antidiuresis and antinatriuresis along with decreases in renal perfusion and glomerular filtration. DA, in doses of $15\;{\mu}g/kg$, also produced antidiuresis and antinaturiuresis. However, after yohimbine-pretreatment DA $15\;{\mu}g/kg$ improved renal hemodynamics, and electrolyte excretion and urine flow rate transiently increased. With $150\;{\mu}g/kg$ DA, the antidiuresis was more marked in the control group. But the yohimbine-pretreated animals responded with marked diuresis and natriuresis, sodium excretion increasing more than three-fold, which lasted for 20 minutes. $K^+-excretion$, osmolar clearance as well as free-water reabsorption increased. Renal hemodynamics improved partly. Apomorphine, a DA agonist, when given icv in doses of $150\;{\mu}g/kg$, produced diuresis and naturiuresis, concomitant with increased renal hemodynamics. Yohimbine-pretreatment however did not abolish the apomorphine-induced diuresis and naturiuresis. Antidiuresis and antinatriuresis elicited by norepinephrine, $10\;{\mu}g/kg$, was not affected by yohimbine-pretreatment. These results indicate that the renal effects of icv DA is not so simple as those of norepinephrine, and the diuretic natriuretic cffect which had been masked by the hemodynamic effect becomes manifest only when the decreases in hemodynamics were removed by the pretreatment of yohimbine. It was further suggested that those DA receptors which mediate the natriuretic response to icv DA is not affected by yohimbine, whereas those receptors involved in the decrease in renal hemodvnamics are blocked by yohimbine. And the possibility of involvement of adrcnergic system in the DA action is not substantiated.
Even though the anticarcinogenic effect of dietary factors especially beta - carotene has been reported by various investigators, the mechanism of the action of ${\beta}-carotene$ has not yet been identified. We carried out the present study to determine the possibilities of relative cancer risk related to dietary intake of vitamin A ( both ${\beta}-carotene$ and retinol ) and blood levels of vitamin A among Koreans. The subjects were divided into two groups; cancer patients and controls. Blood levels for ${\beta}-carotene$ and retinol were analyzed by alumina column chromatography and colorimetry. Dietary intake was examined by food profile and convenient method for evaluating nutritional status through recalling 10 years of food habits. The results obtained are as follows : 1 ) Calorie, protein, fat, and carbohydrate intakes of cancer patient were lower than those of control. Calorie and carbohydrate intakes showed no significant difference but protein and fat intakes were significantly lower in cancer patients. According to cancer sites, in stomach cancer only fat intake was significantly lower than that of control. In lung and larynx cancer calorie, protein, fat and carbohydrate intakes showed similar trend as in control. 2 ) Vitamin A intake of cancer patient was significantly lower than that of control. It was estimated that 83.6% of total Vitamin A intake were provided by ${\beta}-carotene$ for control and cancer patient respectively. 3 ) The mean intake of dietary ${\beta}-carotene$ in cancer patient was significantly lower than that in control ( $7002\;\mu}g/day$ versus $10326\;{\mu}g/day$ ) According to cancer sites in mean intake of dietary ${\beta}-carotene$ was significantly lower in all but stomach cancer compared with that of control. Lung and larynx cancer showed lowest ${\beta}-carotene$ intake with mean value of $5855{\mu}g/day$ and $5492{\mu}g/day$ respectively. 4 ) The mean intake of dietary retinol in cancer patient was significantly lower than that in control ( $245{\mu}g/day$ versus $338{\mu}g/day$ ), but the difference was not significant. 5 ) The relative risk of all cancers in the first (lowest) to the forth quartile level of ${\beta}-carotene$ consumption such as $0-5999{\mu}g/day$. $6000-8999{\mu}g/day$, $9000-11999{\mu}g/day$/ day and $12000-20000{\mu}g/day$ was 85 : 1.7 : 20 : 1.0. The relative risk of all cancers in the first (lowest) to the forth quartile level of retinol consumption, such as $0-299{\mu}g/day$, $300-599{\mu}g/day$, 600-899${\mu}g/day$, and $900-1200{\mu}g/day$, was 1.14 : 067 : 0.21 : 1.0. 6 ) The various food group consumption of cancer patient were significantly lower than those of control in green leafy vegetables, fruits, sea weeds, milk and cheese and eggs. But the Kimchie consumption in cancer patient was three fold higher than that of control ( $1840\;{\mu}g/day$ versus $562\;{\mu}g/day$ ) and in the stomach cancer, Kimchie consumption was the highest, ( $1890\;{\mu}g/day$) According to cancer sites, the consumption of green leafy vegetables was significantly lower in all but stomach cancer compared to control and other vegetables showed no difference between two. In lung and larynx cancer, green leafy vegetables consumption was lowest ( $6094{\mu}g/day$$5921{\mu}g/day$) and milk and cheese consumption was also( $5\;{\mu}g/day$ and $11{\mu}g/day$) 7 ) The recovery of ${\beta}-carotene$ from human serum by alumina column chromatography was $94.4{\pm}2.3%$. 8 ) Cancer patients showed significantly lower serum retinol ($56.4{\pm}18.1\:{\mu}g/100ml$ versus $72.2{\pm}21.8\:{\mu}g/100ml$) and ${\beta}-carotene$ ($48.9{\pm}33.8\:{\mu}g/100ml$ versus $72.2{\pm}42.6\:{\mu}g/100ml$) concentrations than in controls. 9 ) But breast cancer patients were not significantly different from controls in their serum retinol and ${\beta}-carotene$ concentrations.
Kim, Mi Kyeong;Moon, Dong Chul;Hyun, Hye Jin;Kim, Jong-Sik;Choi, Tae Jin;Jung, Sang Bong
Journal of Life Science
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v.26
no.9
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pp.1056-1062
/
2016
Lung cancer is currently the most common malignant disease and the leading cause of mortality in the world and non-small cell lung cancer (NSCLC) accounts for 75-80% of lung cancer cases. miR-155 gene was found to be over expressed in several solid tumors, such as thyroid carcinoma, breast cancer, colon cancer, cervical cancer, pancreatic ductal adenocarcinoma (PDAC) and lung cancer. The aims of this study were to define the expression of miR-155 in lung cancer and its associated clinic-pathologic characteristics. Total RNA was purified from formalin-fixed, paraffin-embedded NSCLC tissues and benign lung tissues. Expression of miR-155 in human lung cancer tissues were evaluated as mean fold changes of miR-155 in cancer tissues compared to benign lung tissues by quantitative real-time reverse transcriptase polymerase chain reaction (real-time qRT-PCR) and associations of miR-155 expression with clinic-pathologic findings of cancer. Compared with the benign control group, miR-155 expression was significantly overexpressed in NSCLCs (p=<0.001). miR-155 was more overexpressed in squamous cell carcinoma than in adenocarcinoma. Poorly differentiated tumors showed significantly overexpression of miR-155 than well-differentiated tumors (p=<0.001). Overexpression of miR-155 was significantly associated with lymph node metastasis (p=<0.05). In survival analysis for all NSCLC patients, high miR-155 expression was significantly correlated with worse overall survival (p=<0.05). These results suggested that miR-155 might play an important role in lung cancer progression and metastasis.
Jang, Ja Yeong;Choi, Yong Ho;Joo, Yoon-Jung;Kim, Hun;Choi, Gyung Ja;Jang, Kyoung Soo;Kim, Chang-Jin;Cha, Byeongjin;Park, Hae Woong;Kim, Jin-Cheol
Research in Plant Disease
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v.21
no.2
/
pp.50-57
/
2015
Control of nematode has become difficult owing to the restricted use of effective soil fumigant, methyl bromide, and other non-fumigant nematicides. Therefore, it is urgently necessary to develop microbial nematicide to replace chemical nematicides. In this study, the 50% aqueous methanol extraction solution of fermentation broths of 2,700 actinomycete strains were tested for their nematicidal activity against second stage of juveniles (J2s) of Meloidogyne incognita. As the results, only the 50% aqueous methanol extraction solution of AN110065, at 20% equivalent to 10% fermentation broth, showed strong nematicidal activity with 78.9% of mortality 24 h after treatment and 94.1% of mortality at 72 h. The 16S rRNA gene sequencing showed that the strain sequence was 99.78% identical to Streptomyces netropsis. The extract of S. netropsis AN110065 fermentation broth was successively partitioned with ethyl acetate and butanol and then the ethyl acetate, butanol and water layers were investigated for their nematicidal activity against the M. incognita. At $1000{\mu}g/ml$, ethyl acetate layer showed the strongest activity of 83.5% of juvenile mortality 72 h after treatment. The pot experiment using the fermentation broth of AN110065 on tomato plant against M. incognita displayed that it evidently suppressed gall formation at a 10-fold diluent treatment. The tomato plants treated with the fermentation broth of S. netropsis AN110065 did not show any phytotoxicity. The results suggest that S. netropsis AN110065 has a potential to serve as microbial nematicide in organic agriculture.
This study aimed to investigate the effects of vocal aerobic treatment (VAT) on the improvement of voice in patients with voice disorders. Twenty patients (13 males, 7 females) were diagnosed with voice disorders on the basis of videostroboscopy and voice evaluations. Acoustic evaluation was performed with the Multidimensional voice program (MDVP) and Voice Range Profile (VRP) of Computerized Speech Lab (CSL), and aerodynamic evaluation with PAS (Phonatory Aerodynamic System). The changes in F0, Jitter, Shimmer, and NHR before and after treatment were measured by MDVP. F0 range and Energy range were measured with VRP before and after treatment, and the changes in Expiratory Volume (FVC), Phonation Time (PHOT), Mean Expiratory Airflow (MEAF), Mean Peak Air Pressure (MPAP), and Aerodynamic Efficiency (AEFF) with PAS. Videostroboscopy was performed to evaluate the regularity, symmetry, mucosal wave, and amplitude changes of both vocal cords before and after treatment. Voice therapy was performed once a week for each patient using the VAT program in a holistic voice therapy approach. The average number of treatments per patient was 6.5. In the MDVP, Jitter, Shimmer, and NHR showed statistically significant decreases (p < .001, p < .01, p < .05). VRP results showed that Hz and semitones in the frequency range improved significantly after treatment (p < .01, p < .05), as did PAS, FVC, and PHOT (p < .01, p < .001). The results for videostroboscopy, functional voice disorder, laryngopharyngeal reflux, and benign vocal fold lesions were normal. Thus, the VAT program was found to be effective in improving the acoustic and aerodynamic aspects of the voice of patients with voice disorders. In future studies, the effect of VAT on the same group of voice disorders should be studied. It is also necessary to investigate subjective voice improvement and objective voice improvement. Furthermore, it is necessary to examine the effects of VAT in professional voice users.
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