• Reproductive and Developmental Biology
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• v.33 no.3
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• pp.171-177
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• 2009

Spermatogonial stem cells(SSCs) only are responsible for the generation of progeny and for the transmission of genetic information to the next generation in male. Other in vitro studies have cultured SSCs for proliferation, differentiation, and genetic modification in mouse and rat. Currently, information regarding in vitro culture of porcine Germline Stem Cell(GSC) such as gonocyte or SSC is limited and is in need of further studies. Therefore, in this study, we report development of a successful culture system for gonocytes of neonatal porcine testes. Testis cells were extracted from $10{\sim}14$-day-old pigs. These cells were harvested using enzymatic digestion, and the harvested cells were purified with combination of percoll, laminin, and gelatin selection techniques. The most effective culture system of porcine gonocytes was established through trial experiments which made a comparison between different feeder cells, medium, serum concentrations, temperatures, and $O_2$ tensions. Taken together, the optimal condition was established using C166 or Mouse Embryonic Fibroblast(MEF) feeder cell, Rat Serum Free Medium(RSFM), 0% serum concentration, $37^{\circ}C$ temperature, and $O_2$ 20% tension. Although we discovered the optimal culture condition for proliferation of porcine gonocytes, the gonocyte colonies ceased to expand after one month. These results suggest inadequate acquirement of ingredients essential for long term culture of porcine GSCs. Consequently, further study should be conducted to establish a successful long-term culture system for porcine GSCs by introducing various growth factors or nutrients.

• Journal of the Korea Convergence Society
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• v.11 no.5
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• pp.35-41
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• 2020

This study was conducted to develop the efficient system for starch hydrolysis suppression using rutin, quercetin and dietary fiber through the statistical mixture design. The three components were replaced with wheat flour at the level of 10% and the mixed gel with three components was characterized by in vitro starch digestion. The mixture design was applied by simplex-centroid experimental model. The quadratic model (R²=0.86) was well fitted and the obtained regression equation indicated that the significant positive effects was observed in the quercetin and fiber mixture. Based on the statistical results, the best mixing ratio of quercetin and fiber was 72: 28 that led to the lowest predicted glycemic index (pGI). Their interactions on the pGI of starch digestibility were clearly visualized in the 3D surface plot. These results suggested that the mixture of quercetin and fiber interact strongly with wheat flour, consequently retarding starch hydrolysis by 15%.

• Animal Bioscience
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• v.34 no.12
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• pp.1940-1950
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• 2021

Objective: The objective of this study was to explore the effects of maize straw treated with calcium oxide (CaO) and various moisture, on the composition and molecular structure of the fiber, and gas production by fermentation in an in vitro rumen environment. Methods: The experiment used 4×3 Factorial treatment. Maize straws were treated with 4 concentrations of CaO (0%, 3%, 5%, and 7% of dry straw weight) and 3 moisture contents (40%, 50%, and 60%). Scanning electron microscopy, Fourier transform infrared spectroscopy and X-ray fluorescence spectroscopy were employed to measure the surface texture, secondary molecular structure of carbohydrate, and calcium (Ca) content of the maize straw, respectively. The correlation of secondary molecular structures and fiber components of maize straw were analyzed by CORR procedure of SAS 9.2. In vitro rumen fermentation was performed for 6, 12, 24, 48, and 72 h to measure gas production. Results: Overall, the moisture factor had no obvious effect on the experimental results. Neutral detergent fiber (NDF), acid detergent fiber, acid detergent lignin, hemicellulose and cellulose contents decreased (p<0.05) with increasing concentrations of CaO treatment. Surface and secondary molecular structure of maize straw were affected by various CaO and moisture treatments. NDF had positive correlation (p<0.01) with Cell-H (H, height), Cell-A (A, area), CHO-2-H. Hemicellulose had positive correlation (p<0.01) with Lignin-H, Lignin-A, Cell-H, Cell-A. Ca content of maize straw increased as the concentration of CaO was increased (p<0.01). Gas production was highest in the group treated with 7% CaO. Conclusion: CaO can adhere to the surface of the maize straw, and then improve the digestibility of the maize straw in ruminants by modifying the structure of lignocellulose and facilitating the maize straw for microbial degradation.

• Plant Biotechnology Reports
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• v.2 no.2
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• pp.145-152
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• 2008

Aerides vandarum and Vanda stangeana are two rare and endangered vandaceous orchids with immense floricultural traits. The intergeneric hybrids were synthesized by performing reciprocal crosses between them. In vitro germination response of the immature hybrid embryos was found to be best on half-strength Murashige and Skoog medium supplemented with 20% (v/v) coconut water/liquid endosperm from tender coconut. Determination of hybridity was made as early as the immature seeds or embryos germinated in vitro, using randomly amplified polymorphic DNA (RAPD) markers. Out of 15 arbitrarily chosen decamer RAPD primers, two were found to be useful in amplification of polymorphic bands specific to the parental species and their presence in the reciprocal crosses. However, a decisive profile that can identify the reciprocal crosses could not be provided by RAPD. Amplification of the trnL-F non-coding regions of chloroplast DNA of the parent species and hybrids aided easy identification of the reciprocal crosses from the fact that maternal inheritance of chloroplast DNA held true for these intergeneric hybrids. Subsequent restriction digestion of the polymerase chain reaction (PCR) amplified trnL-F non-coding regions of chloroplast DNA also consolidated the finding. Such PCR-based molecular markers could be used for early determination of hybridity and easy identification of the reciprocal crosses.

• Restorative Dentistry and Endodontics
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• v.41 no.4
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• pp.283-295
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• 2016

Objectives: In this study, we characterized human dental pulp cells (HDPCs) obtained by different culture methods to establish the most suitable methodology for dental tissue engineering and regenerative endodontic applications. Materials and Methods: HDPCs were isolated by the outgrowth method (HDPCs-OG), the enzymatic digestion method (collagenase/dispase/trypsin, HDPCs-ED), or the combination of both methods (HDPCs-Combined). The expression of mesenchymal stem cell markers (CD105, CD90, and CD73) was investigated. In vitro differentiation capacities of HDPCs into adipogenic, osteogenic, and chondrogenic lineages were compared. Differentiation markers were analyzed by quantitative reverse-transcription polymerase chain reaction (RT-PCR) and western blotting. Results: Our data indicated that whole HDPCs-ED, HPDCs-OG, and HDPCs-Combined could be differentiated into adipogenic, chrondrogenic, and osteogenic cell types. However, we found that the methods for isolating and culturing HDPCs influence the differentiation capacities of cells. HDPCs-OG and HDPCs-ED were preferably differentiated into adipogenic and osteogenic cells, respectively. Differentiation markers shown by RT-PCR and western blotting analysis were mostly upregulated in the treated groups compared with the control groups. Conclusions: Our findings confirmed that cell populations formed by two different culture methods and the combined culture method exhibited different properties. The results of this study could provide an insight into regenerative endodontic treatment using HDPCs.

• Journal of Animal Environmental Science
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• v.15 no.2
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• pp.91-98
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• 2009

Object of this study were to determine the influence of collection times of rumen fluid on in vitro dry matter digestibility (IVDMD) of forage crops. The donor cow was fed concentrate once a day and given free access to grass-legume mixture hay. Main plot was consisted of different collection time of rumen fluid; T1: 1 hour before concentrate feeding, T2: 1 hour after feeding, T3: 4 hour after feeding and T4: 8 hour after feeding. A total of 7 samples of 4 different forage crops (barely, Italian ryegrass, crimson clover, rye) were used for the estimation of IVDMD and tested in three replicates. For the $DAISY^{II}$ incubation, each sample was inserted into each filter bag then heat-sealed and incubated in a digestion vessel for 48 h at $39^{\circ}C$. The times of rumen fluid collection had no significant effect on the IVDMD values over all varieties except for 2 breeds of IRG (Kogreen and Kospeed). IVDMD values with T1 over all varieties were slightly higher than other treatments, however those with both T1 of Kogreen and Kospeed varieties were significantly higher than T2 (p<.05).

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.3
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• pp.392-399
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• 2017

Objective: The objectives were to assess the effects of purified lignin from wheat straw (sodium hydroxide dehydrated lignin; SHDL) on in vitro ruminal fermentation and on the growth performance of feedlot cattle. Methods: In vitro experiments were conducted by incubating a timothy-alfalfa (50:50) forage mixture (48 h) and barley grain (24 h) with 0, 0.25, 0.5, 1.0, and 2.0 mg/mL of rumen fluid (equivalent to 0, 2, 4, 8, and 16 g SHDL/kg diet). Productions of $CH_4$ and total gas, volatile fatty acids, ammonia, dry matter (DM) disappearance (DMD) and digestion of neutral detergent fiber (NDF) or starch were measured. Sixty Hereford-Angus cross weaned steer calves were individually fed a typical barley silage-barley grain based total mixed ration and supplemented with SHDL at 0, 4, 8, and 16 g/kg DM for 70 (growing), 28 (transition), and 121 d (finishing) period. Cattle were slaughtered at the end of the experiment and carcass traits were assessed. Results: With forage, SHDL linearly (p<0.001) reduced 48-h in vitro DMD from 54.9% to 39.2%, NDF disappearance from 34.1% to 18.6% and the acetate: propionate ratio from 2.56 to 2.41, but linearly (p<0.001) increased $CH_4$ production from 9.5 to 12.4 mL/100 mg DMD. With barley grain, SHDL linearly increased (p<0.001) 24-h DMD from74.6% to 84.5%, but linearly (p<0.001) reduced $CH_4$ production from 5.6 to 4.2 mL/100 mg DMD and $NH_3$ accumulation from 9.15 to $4.49{\mu}mol/mL$. Supplementation of SHDL did not affect growth, but tended (p = 0.10) to linearly reduce feed intake, and quadratically increased (p = 0.059) feed efficiency during the finishing period. Addition of SHDL also tended (p = 0.098) to linearly increase the saleable meat yield of the carcass from 52.5% to 55.7%. Conclusion: Purified lignin used as feed additive has potential to improve feed efficiency for finishing feedlot cattle and carcass quality.

• Journal of Nutrition and Health
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• v.29 no.8
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• pp.861-866
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• 1996

Tannins in plant foods and beverages may produce antinutritional or toxic effects although some proteins with high affinity for tannins seem to function as defense mechanism to tannin toxicity. Our objectives were to investigate of tea tannins, iron and protein and to evaluate the role of proteins in tannin effects on iron solubility. Iron solubility in vitro was measured using tea with and without proteins. Mixtures of tea, protein in varying concentrations(either gelatin or bovine serum albumin), and iron(eithe 10 or 50ug/mL) were prepared. Controls contained water in place of tea. Iron bioavailability was assessed by measuring iron solubility in the simulated gastric condition with pepsin digestion. Bound iron was removed by centrifugation and soluble in tea alone. When iron concentratin was 10ug/mL, addition of small amounts of protein to tea dramatically reduced iron solubility, but solubility of iron increased in the tea mixturea as the concentration of protein was increased. The percnetage of iron that precipitated was much greater at 10ug Fe/mL than the values at 50ug Fe/mL suggesting that the iron binding sites on the tea-protein complex was saturated. These results suggest that interactions of iron with tea tannins are influenced by the concentratins of protein and iron.

• Korean journal of food and cookery science
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• v.19 no.1
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• pp.107-113
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• 2003

In a previous study, a dietary supplement was developed in our lab using natural herbal extracts against digest enzyme activity in GI tract for weight control. This natural herbal extracts could regulate absorption of glucose and lipid by the inhibition of digest enzyme activity. In this study, we screened the natural herbs that inhibit glucoamylase activity and developed an water extract of cinnamon. The cinnamon extract delayed and decreased the increment of carbohydrate degradation through the inhibition of glucoamylase activity in vitro. Fifty volunteers were subjected to the intake of the herbal extracts by taking twice a day for 60 days. As a result, the treated subjects lost 3 kg of body weight and 3.5 kg of body fat mass after the treatment. Furthermore, the body mass index and waist size were significantly decreased during the experimental period. Above results suggested that the administration of the dietary additives composed of cinnamon and natural herbal extract improves the obesity by the decrement of body weight and body fat mass.

• Journal of Microbiology and Biotechnology
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• v.32 no.5
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• pp.657-662
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• 2022

Glycosyltransferase (GT)-specific degenerate PCR screening followed by in silico sequence analyses of the target clone was used to isolate a member of family1 GT-encoding genes from the established fosmid libraries of soil actinomycetes Micromonospora echinospora ATCC 27932. A recombinant MeUGT1 was heterologously expressed as a His-tagged protein in E. coli, and its enzymatic reaction with semi-synthetic phenoxodiol isoflavene (as a glycosyl acceptor) and uridine diphosphate-glucose (as a glycosyl donor) created two different glycol-attached products, thus revealing that MeUGT1 functions as an isoflavonoid glycosyltransferase with regional flexibility. Chromatographic separation of product glycosides followed by the instrumental analyses, clearly confirmed these previously unprecedented glycosides as phenoxodiol-4'-α-O-glucoside and phenoxodiol-7-α-O-glucoside, respectively. The antioxidant activities of the above glycosides are almost the same as that of parental phenoxodiol, whereas their anti-proliferative activities are all superior to that of cisplatin (the most common platinum chemotherapy drug) against two human carcinoma cells, ovarian SKOV-3 and prostate DU-145. In addition, they are more water-soluble than their parental aglycone, as well as remaining intractable to the simulated in vitro digestion test, hence demonstrating the pharmacological potential for the enhanced bio-accessibility of phenoxodiol glycosides. This is the first report on the microbial enzymatic biosynthesis of phenoxodiol glucosides.