• Journal of Pharmaceutical Investigation
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• 제26권2호
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• pp.91-98
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• 1996

Precipitation was formed during the preparation of decoction from a mixure of Scutellariae Radix and Coptidis Rhizoma or Phellodendri Cortex according to the prescription of Hwang-ryean-hae-dog-tang. Baicalin and berberine, the active ingredients of the two herbal medicine were identified in coprecipitated product. Pills were prepared using the coprecipitated product and various binders. The dissolution rate of baicalin and berberine from pills was increased in at pH1.2 when acacia or tragacanth was used. The absorption rate of baicalin from the coprecipitated product was faster than that from Scutellaria extract, but the absorption of berberine from CPP was slower in stomach, duodenum and jejunum of rats compared with Coptis extract. The time required for the maximum serum concentration (Cmax) of baicalin and berberine from CPP in mice were 150 and 200 min after oral administration, respectively. The maximum serum concentration of baicalin from CPP in mice was higher than Scutellaria extract, but the concentration of berberine was lower compared with Coptis extract. The minimum inhibitory concentration of CPP was below $50\;{\mu}g/ml$ against gram positive bacteria, and was higher than that against gram negative bacteria. The antibacterial activity of CPP was lower than that of herberine, but was more potent than Scutellaria extract. It was found that the inhibition rates of growth by CPP against S. epidermidis, K. pneumoniae, B. cereus and S.aureus were 60.0, 51.1, 45.4 and 39.9%, respectively.

• Journal of Animal Science and Technology
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• 제64권6호
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• pp.1105-1116
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• 2022

Recently, we reported the robust in vitro three-dimensional (3D) expansion of intestinal organoids derived from adult bovine (> 24 months) samples. The present study aimed to establish an in vitro 3D system for the cultivation of intestinal organoids derived from growing cattle (12 months old) for practical use as a potential alternative to in vivo systems for various purposes. However, very few studies on the functional characterization and 3D expansion of adult stem cells from livestock species compared to those from other species are available. In this study, intestinal crypts, including intestinal stem cells, from the small intestines (ileum and jejunum) of growing cattle were isolated and long-term 3D cultures were successfully established using a scaffold-based method. Furthermore, we generated an apical-out intestinal organoid derived from growing cattle. Interestingly, intestinal organoids derived from the ileum, but not the jejunum, could be expanded without losing the ability to recapitulate crypts, and these organoids specifically expressed several specific markers of intestinal stem cells and the intestinal epithelium. Furthermore, these organoids exhibited key functionality with regard to high permeability for compounds up to 4 kDa in size (e.g., fluorescein isothiocyanate [FITC]-dextran), indicating that apical-out intestinal organoids are better than other models. Collectively, these results indicate the establishment of growing cattle-derived intestinal organoids and subsequent generation of apical-out intestinal organoids. These organoids may be valuable tools and potential alternatives to in vivo systems for examining host-pathogen interactions involving epithelial cells, such as enteric virus infection and nutrient absorption, and may be used for various purposes.

• 한국식품영양과학회지
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• 제45권6호
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• pp.783-788
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• 2016

본 연구는 수수의 섭취가 대표적인 트랜스지방산인 elaidic acid, linoelaidic acid와 cholesterol의 소장흡수에 어떤 영향을 미치는지를 조사하기 위하여 설계하였다. 소장의 지방흡수를 in vivo 상태에서 측정하기 위해 소장흡수율 측정모델을 이용하여 십이지장주입관과 림프채취관을 삽입하는 미세수술을 진행하였다. 실험동물의 십이지장으로 $146.4{\mu}mol$ trielaidin, $36.8{\mu}mol$ trilinoelaidin, $452.0{\mu}mol$ triolein, $1.0{\mu}Ci$ $^{14}C-cholesterol$, $20.7{\mu}mol$ cholesterol, $396.0{\mu}mol$ Na-taurocholate, PBS buffer(pH 6.4)가 포함된 지질유화액만 공급받는 동물군을 대조군, 수수추출물이 혼합된 지질유화액을 공급받는 동물군을 수수군으로 설계하였다. 지질유화액은 시간당 3 mL씩 8시간 동안 주입하면서 림프관으로 흡수되는 elaidic acid, linoelaidic acid와 각종 지방산 그리고 $^{14}C-cholesterol$의 양을 분석하여 수수에 의한 영향을 비교하였다. 먼저 8시간 동안 분비된 림프액의 양은 대조군과 수수군 간에 유의적인 차이를 나타내지 않았다. 하지만 8시간 동안의 elaidic acid 흡수는 수수추출물의 공급으로 유의적으로 억제되는 것으로 나타났다. Linoelaidic acid 또한 마찬가지로 수수추출물 공급으로 유의적으로 그 흡수가 억제된 것으로 나타났다. 트랜스지방산을 비롯해 palmitic acid, stearic acid, oleic acid, linoleic acid, arachidonic acid, docosahexaenoic acid, $^{14}C-cholesterol$ 또한 대조군보다 수수군에서 흡수가 유의적으로 억제된 것으로 나타났다. 이상의 결과들은 수수추출물이 트랜스지방산과 cholesterol의 소장 흡수를 억제할 수 있다는 사실을 증명한다. 이번 연구를 바탕으로 수수 섭취가 트랜스지방이나 cholesterol의 흡수를 농도 비례적으로 억제할 수 있을지, 소화흡수 과정 중 정확히 어떤 과정에 영향을 미쳤는지에 관한 연구가 앞으로 진행될 과제라고 판단된다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권4호
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• pp.554-562
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• 2006

Two lines of mice, M16 selected for rapid growth and a randomly selected control ICR as well as their reciprocal crosses were used to study the effects of genotype on whole-body energetics and intestinal responses to monensin. Six mice, eight weeks of age, from each line or reciprocal cross were assigned to one of two treatments, 1) drinking water containing 20 mmol/L monensin dissolved in 0.5% V/V ethanol, and 2) drinking water containing 0.5% V/V ethanol (control) for two weeks. After 11 days (age of 9 weeks and 4 days), whole-body $O_2$ consumption was measured. At the end of two weeks, jejunal $O_2$ consumption, intestinal tissue composition and histomorphometrics as well as the rate and efficiency of glucose absorption were estimated. In comparison with the control, monensin administration in drinking water resulted in less daily water intake (13.4 vs. 15.5 ml/mouse, p<0.01), less protein to DNA ratio of jejunal mucosa (5.41 vs. 6.01 mg/mg, p<0.05), lower villus width (88 vs. $100{\mu}m$, p<0.05), and less jejunal tissue $O_2$ consumption enhancement by alcohol (7.2 vs. 10.5%, p<0.01) in mice. Other than those changes, monensin had little (p>0.05) effect on variables measured in either line of mice or their reciprocal cross. In contrast, the M16 line, selected for rapid growth, as compared to the ICR controls or the reciprocal crosses, had less initial (pre-monensin treatment) whole-body $O_2$ consumption per gram of body weight (1.68 vs. $2.11-2.34{\mu}mol/min{\cdot}g$ BW, p<0.01) as compared to the ICR and reciprocal crosses. In addition, the M16 mice exhibited greater growth (412 vs. 137-210 mg/d, p<0.05), better feed efficiency (41.7 vs. 19.9-29.3 mg gain/g feed, p<0.05), shorter small intestines adjusted for fasted body weight (1.00 vs. 1.22-1.44 cm/g FBW, p<0.05), wider villi (109 vs. $87-93{\mu}m$, p<0.05), more mature height of enterocytes (28.8 vs. $24.4-25.1{\mu}m$, p<0.05) and a lower rate (91 vs. $133-145{\eta}mol\;glucose/min{\cdot}g$ jejunum, p<0.05) and less energetic efficiency (95 vs. $59-72{\eta}mol$ ATP expended/${\eta}mol$ glucose uptake, p<0.05) of glucose absorption compared to the ICR line and the reciprocal cross. Monensin had little (p>0.05) effect on whole-body $O_2$ consumption and jejunal function, whilst selection for rapid growth resulted in an apparent down-regulation of intestinal function. These data suggest that genetic selection for increased growth does not result in concomitant changes in intestinal function. This asynchrony in the selection for production traits and intestinal function may hinder full phenotypic expression of genotypic growth potential.

• Archives of Pharmacal Research
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• 제25권1호
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• pp.86-92
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• 2002

The enhancement of paracellular transport of heparin disaccharide using several absorption enhancers across Caco-2 cell monolayers was tested . The cytotoxicity of these enhancers was also examined. The enhancing effects by Quillaja saponin, diponin glycyrrhizinate, $18{\beta}-glycyrrhetinic$ acid, sodium caprate and taurine were determined by changes in transepithelial electrical resistance (TEER) and the amount of heparin disaccharide transported across Caco-2 cell monolayers. Among the absorption enhancers, $18{\beta}-glycyrrhetinic$ acid arid taurine decreased TEER and increased the permeability of heparin disaccharide in a dose-dependent and time-dependent manner with little or negligible cytotoxicity. Our results indicate that these absorption enhancers can widen the tight junction, which is a dominant paracellular absorption route of hydrophilic compounds . It is highly possible that these absorption enhancers can be applied as pharmaceutical excipients to improve the transport of macromolecules and hydrophilic drugs having difficulty in permeability across the intestinal epithelium.

• Food Science and Biotechnology
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• 제17권3호
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• pp.564-568
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• 2008

The effects of sodium copper chlorophyllin (SCC) on bioaccessibility and uptake of mercury from fish were investigated using an in vitro digestion coupled with a Caco-2 cell. Fish along with SCC was subjected to a simulated in vitro digestion, which simulates both the gastric and small intestinal phase in vivo. Mercury bioaccessibility, the amount of mercury released from fish to aqueous phase following a digestion, was measured. Various amounts of SCC (0.1-25 mg) significantly reduced mercury bioaccessibility in a dose dependent manner by 49-89% compared to the negative control (fish without SCC) (p<0.05). Mercury bioaccessibility in varying molar ratios of mercury to positive control, 2,3-dimercapto-1-propane sulfonate (DMPS) was between 24 and 52%. Mercury uptake by Caco-2 cells from test media containing aqueous phase following in vitro digestion was measured after 6 hr incubation at $37^{\circ}C$. Cellular mercury uptake with increasing amount of SCC ranged from 0.352 to $0.052\;{\mu}g$ mercury/mg protein, while those in DMPS treatment were between 0.14 and $0.27\;{\mu}g$ mercury/mg protein. Our study suggests that SCC can reduce mercury absorption following fish consumption and may be efficient as a synthetic chelating agent for long term chronic mercury exposure in fish eating populations.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제18권4호
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• pp.230-237
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• 2015

Purpose: Various gastrointestinal factors may contribute to maladaptive behavior in children with autism spectrum disorders (ASD). To determine the association between maladaptive behavior in children with ASD and gastrointestinal symptoms such as severity, intestinal microbiota, inflammation, enterocyte damage, permeability and absorption of opioid peptides. Methods: This observational cross-sectional study compared children with ASD to healthy controls, aged 2-10 years. Maladaptive behavior was classified using the Approach Withdrawal Problems Composite subtest of the Pervasive Developmental Disorder Behavior Inventory. Dependent variables were gastrointestinal symptom severity index, fecal calprotectin, urinary D-lactate, urinary lactulose/mannitol excretion, urinary intestinal fatty acids binding protein (I-FABP) and urinary opioid peptide excretion. Results: We did not find a significant difference between children with ASD with severe or mild maladaptive behavior and control subjects for gastrointestinal symptoms, fecal calprotectin, urinary D-lactate, and lactulose/mannitol ratio. Urinary opioid peptide excretion was absent in all children. Children with ASD with severe maladaptive behavior showed significantly higher urinary I-FABP levels compared to those with mild maladaptive behavior (p=0.019) and controls (p=0.015). Conclusion: In our series, maladaptive behavior in ASD children was not associated with gastrointestinal symptoms, intestinal inflammation (no difference in calprotectin), microbiota (no difference in urinary D-lactate) and intestinal permeability (no difference in lactulose/manitol ratio). ASD children with severe maladaptive behavior have significantly more enterocyte damage (increased urinary I-FABP) than ASD children with mild maladaptive behavior and normal children.

• Journal of Pharmaceutical Investigation
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• 제35권2호
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• pp.111-116
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• 2005

The unstirred water layer (UWL), which has been known to exist in the boundary of the intestinal lumen and intestinal wall, often behaves as an absorption barrier especially for lipophilic drugs. The intestinal absorption of drugs is often characterized using Caco-2 cell monolayers grown on Transwell polycarbonate membranes. The permeability $(P_{app})$ of drugs across the cell monolayer might be influenced by the agitation of the donor compartment, since the width of UWL on the surface of the cell monolayer would be reduced by the agitation. In this study, the effect of agitation of the donor compartment with 60 rpm on the permeability was measured for 12 drugs with a wide range of lipophilicity and permeability. The $P_{app}$ of mannitol, tributylmethyl ammonium, cimetidine, ranitidine, hydrocortisone, benzylpenicillin and loxoprofen was not influenced by the agitation, while the $P_{app}$ of theophylline, propranolol, YH439, phenylpropanolamine and testosterone was increased by the agitation. There was a significant correlation between the increase of $P_{app}$ by agitation and the lipophilicity for the compounds having $P_{app}>2{\times}10^{-5}$ cm/sec. No correlation was observed for the difference in $P_{app}$ by agitation and the molecular weight, or lipophilicity of the drugs. Therefore, the agitation rate of the donor compartment in the Caco-2 cell monolayer study should be carefully controlled in order to estimate $P_{app}$ reproducibly especially for lipophilic drugs.

• 약학회지
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• 제49권2호
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• pp.185-191
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• 2005

The absorption of a P-gp substrate, rhodamine 123, from a liposomal dosage form was investigated across Caco-2 cell monolayers, rat intestines and rat intestinal Peyer's patches in Ussing chamber, Rhodamine 123 was incorporated into liposomes according to the standard evaporation method, which led to a production of liposomes with a mean diameter of 71.3 nm. The permeability (Papp of rhodamine 123 from a water solution across the monolayer was $2.45{\times}10^{-6}$ cm/s for $A{\leftrightarrow}B$ (apical to basal) and $14.0{\times}10^{-6}$ cm/s for $B{\leftrightarrow}A$ (basal to apical) directions, consistent with the fact that rhodamine 123 is one of the P-gp substrates. The transport of rhodamine 123 from the liposomal dosage form was much lower for both directions compared to the solution of rhodamine 123. The transport of rhodamine 123 across the rat intestine was also significantly decreased for both directions, I.e., influx and efflux, by the liposomal incorporation of the compound. The transport of rhodamine 123 across the Peyer's patch was substantially reduced by liposomal incorporation. No difference was found in the transport between the Peyer's patch and non-Peyer's patch. These observations suggest that the contribution of transport via Peyer's patches in the uptake of liposomes may be minimal, especially for rapidly absorbed compounds like rhodamine 123. Therefore, the increased absorption of P-gp substrates does not appear to be feasible by incorporating the compounds in liposomes, due to negligible involvement of Peyer's patches in the uptake of particulate dosage forms like liposomes. Liposomes may rather represent a sustained release dosage form of incorporated compounds.

• Journal of Pharmaceutical Investigation
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• 제40권spc호
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• pp.19-31
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• 2010

Orally administrated drugs permeate the biological membrane by various transport mechanisms. The oral absorption potential is closely related to the physicochemical properties of the drug and interaction with the physiological factors surrounding the site of absorption. Assessment of the drug membrane permeability is an integral part of the early stage drug developmental process. Appropriate selection of the permeability screening method at the right stage of drug development process is important in achieving successful developmental outcomes. This review aims at introducing currently available in vitro and in vivo screening methods for the membrane permeability assessment.