Consecutive Oral Administration Research Articles

Pharmacokinetics, pharmacodynamics and safety of 15mg-tolvaptan administered orally for 7 consecutive days to Chinese patients with child-Pugh B cirrhosis.

Background: Tolvaptan, a selective vasopressin V2-receptor antagonist, can elicit a diuretic effect without significant electrolyte loss. The aims were to evaluate multiple-dose pharmacokinetics, pharmacodynamics and safety of daily administration of 15mg tolvaptan in Chinese adult patients with confirmed Child-Pugh Class B cirrhosis accompanied by ascites. Methods: This was an open-label, single-center, single- and multiple-dose study. All patients received a daily 15mg dose of tolvaptan for 7 consecutive days. The plasma concentrations of tolvaptan and its two metabolites (DM-4103, DM-4107) were measured using high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). In addition, various pharmacokinetics parameters were calculated. The pharmacodynamic outcomes evaluated changes in serum sodium and potassium concentrations, daily urine volume, daily water consumption, fluid balance and body weight. Safety profiles, including the incidence of treatment-emergent adverse events (TEAEs), were carefully recorded. Results: Eleven patients with Child-Pugh B cirrhosis were eventually enrolled in the study. Plasma concentrations of tolvaptan and DM-4107 reached steady-states after 7 days of consecutive oral administration. No accumulation of tolvaptan or DM-4107 was found, but DM-4103 accumulated 18.2-fold after multiple-dosing. The daily urine volume and daily water consumption were statistically significantly increased after administration of tolvaptan from Day 1 to Day 7 (all p < 0.05), accompanied by an increased serum sodium concentration. Of 11 patients, 9 (81.8%) reported 20 TEAEs, with the majority being mild to moderate in severity. The most commonly occurring TEAEs were thirst (45.5%), pollakiuria (36.4%) and dry mouth (27.3%). Conclusion: Tolvaptan at a daily dose of 15mg had a diuretic effect but did not increase serum sodium excretion or lead to tolvaptan accumulation. It is therefore can be safely used for short-term treatment of Chinese adult patients with confirmed Child-Pugh B cirrhosis. Clinical Trial Registration: https://clinicaltrials.gov/search?term=NCT01359462, identifier NCT01359462.

Tissue residue distribution and withdrawal time estimation of trimethoprim and sulfachloropyridazine in Yugan black-bone fowl (Gallus gallus domesticus Brisson)

Black-bone fowl are different from ordinary broilers in appearance and are considered to have rich nutritional properties. However, the metabolism of therapeutic drugs in black-bone fowl remains unclear. This study aimed to determine the tissue residue depletion kinetics of trimethoprim and sulfachloropyridazine in Yugan black-bone fowl, after daily oral administrations for 5 days at 4 mg/kg bw/day trimethoprim and 20 mg/kg bw/day sulfachloropyridazine, and to calculate the withdrawal times. After consecutive oral administrations, the tissues (liver, kidney, muscle and skin/fat) were collected at each of the following time points (0.16, 1, 3, 5, 7, 9, 20, 30 and 40 days). A newly-devised LC-MS/MS method was used to analyse the concentrations of trimethoprim and sulfachlorpyridazine in target tissues. The results showed that sulfachloropyridazine was rapidly metabolised in broilers, and there was no residue in all tissues 3 days post-administration. The concentration of trimethoprim in black-bone fowl skin/fat is the highest, and its metabolism rate is low. After 40 days, the concentration of trimethoprim in skin/fat is still as high as 140.1 ± 58.0 μg/kg, exceeding the maximum residue limit. In order to protect consumers’ health, it is suggested that the withdrawal time of TMP in Yugan black-bone fowl is 69 days.

Neurobehavioral effects of cinnabar and the cinnabar-containing pediatric prescription, Yi-Nian-Jin, in juvenile rats

BackgroundCinnabar, a mercury-containing mineral medicine, has long been widely used in pediatric prescriptions. The safety of cinnabar-containing prescriptions, particularly for children, is drawing increasing attention worldwide. However, whether cinnabar and these pediatric prescriptions have adverse effects on neurobehavior is unknown. Yi-Nian-Jin (YNJ), a classic pediatric prescription, contains 5.66% (w/w) cinnabar, along with other four herbs. YNJ is widely prescribed to promote digestion, eliminate phlegm, and prevent constipation in children (aged 0–6 years). In this study, we used YNJ as an example of cinnabar-containing pediatric prescriptions to determine mercury absorption, distribution, and accumulation and further investigate its potential neurotoxicity in juvenile rats. Material and methodsLow (67.9 mg/kg), middle (169.8 mg/kg), and high dose (339.6 mg/kg) of cinnabar, and low (1.2 g/kg), middle (3.0 g/kg), and high dose (6.0 g/kg) of YNJ were used in this study, corresponding to 3, 7.5, and 15 times the clinically equivalent dose, respectively. Juvenile rats were orally administered different doses of cinnabar or YNJ for 14 consecutive days. The mercury content in rat blood and tissues (brain, liver, and kidney) and serum biochemical changes on day 14 of consecutive administration and on day 14 after cessation were measured. Moreover, a series of behavioral assays (open field, elevated plus-maze, and Morris water maze assays) were performed after 14 consecutive days of administration. ResultsThe mercury absorption, distribution, and accumulation of cinnabar and YNJ in juvenile rats were substantially different. Mercury in cinnabar was absorbed to a greater extent than that in YNJ, and the mercury content in cinnabar high-dose group (cinnabar-H) was approximately seven times higher than that in YNJ high-dose group (YNJ-H) on day 14 of administration. In contrast, compared with that of cinnabar, the mercury content in YNJ accumulated more in the tissues, especially in the brain and kidney. Repeated administration of cinnabar or YNJ did not affect liver function, renal function, learning, and memory in juvenile rats. However, repeated administration of YNJ at a high dose (6.0 g/kg) affected locomotor activity in juvenile rats. Repeated administration of cinnabar (339.6 mg/kg) or YNJ (>1.2 g/kg) induced anxiety-related behavior in juvenile rats. ConclusionsMercury in YNJ exhibited lower absorption but higher accumulation in tissues than those of the mercury in cinnabar. Consecutive oral administration of cinnabar or YNJ had no impact on liver function, renal function, learning, and memory, but could cause motor dysfunction and anxiety in juvenile rats.

Establishment of Optimal Conditions to Extract Bioactive Substances from Gamazumi Using Supercritical Carbon Dioxide

The fruits of Gamazumi (Viburnum dilatatum THUNB) have been previously reported to suppress the adverse effects of oxidative stress in rats. To reduce time and cost, we attempted to establish optimal conditions for extracting bioactive components from Gamazumi fruits using supercritical carbon dioxide. To verify whether the conventional hexane extraction may be replaced by the supercritical carbon dioxide extraction, component analysis results and antigenotoxic potential in mice were used to compare bioactivity of the supercritical carbon dioxide and hexane extracts. &#x0D; At the same extraction pressure, the extraction rate was maximized when supercritical carbon dioxide with a low temperature was used, and extraction efficiency was improved. GC/MS analysis revealed that vitamins E and stigmasterols were included in supercritical carbon dioxide and hexane extracts, and that no qualitative differences between supercritical carbon dioxide and hexane extracts were observed. The antigenotoxic potential of Gamazumi extracts was studied in mice exposed to cigarette smoke inhalation. Mice received single or 5 consecutive oral administrations of Gamazumi extracts at 0, 3, and 6 h prior to smoke inhalation. Although single administration decreased nuclei tail length in the stomach when both administration and the intervals of smoke inhalation were short, five consecutive administrations decreased tail length in the lung and stomach regardless of the interval. At short intervals, inhaled cigarette smoke and orally administered extracts may be simultaneously present in the gastric cavity, and direct reaction between cigarette smoke and extract is possible. The antioxidant activity of Gamazumi extracts may result in antigenotoxic potential. There were no differences in component analysis and antigenotoxic potential between supercritical carbon dioxide and hexane extracts of Gamazumi; thus, it is possible to replace the conventional hexane extraction with the supercritical carbon dioxide extraction.

Withdrawal Interval Estimation of Doxycycline in Yellow Catfish (Pelteobagrus fulvidraco) Using an LC-MS/MS Method Based upon QuEChERS Sampling Preparation.

Limited information on residue depletion of doxycycline (DC) is available in yellow catfish (Pelteobagrus fulvidraco) using the determination of LC-MS/MS based upon a rapid and simple method of sample preparation. This study collected plasma and tissue samples of yellow catfish at pre-determined time points following 3-day consecutive oral administration of DC at 20 mg/kg. The samples were prepared using a QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) approach. The limit of detection and the limit of quantitation were 2.5 and 5 µg/kg or µg/L, respectively, for DC in plasma and tissues (e.g., muscle + skin, liver, kidney, and gill). The recoveries of DC ranged from 67.2% to 86.2%. The decision limit (CCα) and the detection capability (CCβ) were from 106.2 to 127.8 μg/kg or μg/L. The withdrawal times of DC in muscle + skin were estimated to be 22 d based on the guidelines in China and Europe and 27 d based on Japan’s standard. Overall, this study not only provides an efficient method to rapidly determine the DC concentrations in fish-derived tissues but also provides important information on the safety assessment of DC in aquatic animal-derived food products.

Oral Administration of Probiotic Bifidobacterium breve Improves Facilitation of Hippocampal Memory Extinction via Restoration of Aberrant Higher Induction of Neuropsin in an MPTP-Induced Mouse Model of Parkinson's Disease.

We previously reported that 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced Parkinson’s disease (PD) model mice (PD mice) facilitate hippocampal memory extinction, which may be the cause of cognitive impairment in PD. Recent studies on the consumption of probiotics have reported a variety of beneficial effects on the central nervous system via the microbiota–gut–brain axis. In this study, we investigated the effects of oral administration of Bifidobacterium breve strain A1 [MCC1274] (B. breve A1) on the facilitation of hippocampal memory extinction observed in PD mice. We found that four-day consecutive oral administration of B. breve A1 restored facilitation of contextual fear extinction in PD mice. Hippocampal mRNA expression levels of postsynaptic density protein-95 and synaptophysin significantly decreased in the PD mice, but mRNA and protein expression levels of neuropsin increased. Furthermore, CA1 apical spine density was significantly reduced in PD mice. On the other hand, administration of B. breve A1 to PD mice recovered all these expression levels and the CA1 spine density to control levels. These results suggest that increased induction of neuropsin is involved in abnormal changes in hippocampal synaptic plasticity, and that B. breve A1 imposes reins on its expression, resulting in the restoration of abnormal hippocampal synaptic plasticity and the facilitation of fear extinction in PD mice.

Protamine and BSA–dextran complex emulsion improves oral bioavailability and anti-tumor efficacy of paclitaxel

Food protein and polysaccharide complex emulsions are safe carriers of hydrophobic drugs and nutrients. To improve oral bioavailability and therapeutic/healthy efficacy of hydrophobic drugs and nutrients, herein, protamine (PRO), a cationic cell-penetrating peptide, was introduced into protein and polysaccharide complex emulsion. The electrostatic complex of PRO and BSA–dextran conjugate (BD) produced by Maillard reaction was used as emulsifier to produce oil-in-water emulsion (@BD/PRO). The BSA molecules were crosslinked at the oil–water interface by a heat treatment and the PRO chains were simultaneously anchored in the interface. BD emulsion (@BD) without PRO was produced for comparation. Paclitaxel (PTX), a hydrophobic antineoplastic drug, was encapsulated in the emulsions with 99% loading efficiency and 6.4% loading capacity. The emulsions had long-term stability. The bioavailability and H22 tumor inhibition efficacy of PTX@BD/PRO were 40% and 70% higher than those of PTX@BD, respectively, after oral administration in the mice. More importantly, orally administrated PTX@BD/PRO had the same anti-tumor efficacy as intravenously injected commercial PTX injection. No abnormality was observed in the main organs of the mice after consecutive oral administration of PTX@BD/PRO. This study indicates that @BD/PRO is an excellent carrier of hydrophobic drugs/nutrients and is suitable for long-term oral administration.

Innocuity and antioxidant activities of &lt;i&gt;Pentadesma butyracea&lt;/i&gt; (1824) leaves for its use in hormone replacement therapy

Pentadesma butyracea is a tall tree of Western Africa commonly called better tree. Leaves are used in the treatment of several diseases including genitourinary, pregnancy and metabolic disorders. The aim of this study was to assess toxicity and antioxidant activity of hydroethanolic extract of P. butyracea leaves. The major compounds of the extract were previously identified. Oral acute toxicity was achieved on NMRI mice during 72 h after single oral-administration whereas oral sub-acute toxicity was achieved on Wistar rats during 28-days. Antioxidant activity was assessed through DPPH, FRAP and lipid peroxidation inhibition. The major phytochemicals identified in the hydroethanolic extract of P. butyracea leaves were tannins, steroids and triterpenoids, anthocyanosids, anthraquinones, saponosids, mucilage and gums. The acute toxicity study showed that the extract did not cause neither sign of toxicity nor mortality until 3000 mg/kg. The subacute toxicity study showed that the extract did not cause signs of toxicity after 28-days consecutive oral administration. Nevertheless, platelets count. total cholesterol and AST level were significantly (p&lt;0.05) reduced. Assessment of antioxidant activity revealed that the extract possessed a great potential of scavenging free radical, to inhibit lipid peroxidation and to reduce ferric power. The extract could have hepaprotective activity and may be benefit for cholesterol reduction.Keywords: Pentadesma butyraceae, toxicity, phytochemistry, antioxidant

An Oral Vaccination Method with the Aid of Capsaicin against Viral Nervous Necrosis (VNN)

To develop an oral vaccination method for viral nervous necrosis (VNN) caused by piscine nodavirus (betanodavirus), sevenband grouper Epinephelus septemfasciatus was immunized with inactivated redspotted grouper nervous necrosis virus (RGNNV) vaccine. ​Seven-day consecutive oral administration of the RGNNV vaccine via feed failed to induce protection in fish against the virus challenge at 21 days post-immunization. ​However, anal intubation of the vaccine conferred protective immunity on fish, although virus neutralizing antibodies could not be detected. Similar protective immunity was also confirmed by oral administration of vaccine via feed supplemented with capsaicin, suggesting that capsaicin modulated the intestinal mucosal barrier to induce immune response.

Preventive effects of Eucommia ulmoides leaf extract and its components on UVB-induced immunosuppression in mice

Ultraviolet (UV) irradiation is a well known photo-immunosuppressant. To determine the effect of Eucommia ulmoides leaf extract (ELE) components (chlorogenic acid, geniposidic acid, and asperuloside) on UVB induced immunosuppression, we examined mice that were contact hypersensitized. Systemic immunosuppression was induced in the C57BL/6j mice that were sensitized with 0.5% oxazolone through the skin by a single exposure to UVB. Consecutive oral administration of ELE, chlorogenic acid, or geniposidic acid significantly reduced UVB-induced suppression of the contact hypersensitivity (CHS) response. The treatment with chlorogenic acid or geniposidic acid decreased the serum interleukin-10 levels in the sensitized mice after UVB irradiation. Additionally, the level of plasma reactive oxygen species and the expression of skin mast cells decreased in the chlorogenic acid-treated mice. Contrarily, the expression of regulatory T cell in the skin decreased in the geniposidic acid-treated mice. These results suggest that chlorogenic and geniposidic acids prevent the photo-immunosuppression of the CHS response.

Anti-obesity effects of yellow catfish protein hydrolysate on mice fed a 45% kcal high-fat diet.

Obesity contributes to the etiologies of a variety of comorbid conditions, such as type 2 diabetes, hypertension and cardiovascular disease. In the present study, the anti-obesity effects of yellow catfish protein hydrolysate (YPh) were observed in mice fed a 45% kcal high-fat diet (HFD) compared with those of mice treated with simvastatin. The HFD-fed control mice exhibited noticeable increase in body weight, and whole-body and abdominal fat densities, periovarian and abdominal wall-deposited fat pad weight, as well as in the levels of triglycerides (TG), blood total cholesterol (TC), low-density lipoprotein, alanine aminotransferase, aspartate aminotransferase, creatinine, blood urea nitrogen, and in the fecal TG and TC contents. However, they exhibited a decrease in serum high-density lipoprotein levels. In addition, an increase was detected in periovarian and dorsal abdominally deposited fat pad thickness, adipocyte hypertrophy, the number of steatohepatitis regions, hepatocyte hypertrophy and lipid droplet deposition-related renal tubular vacuolation degenerative lesions, along with increased hepatic lipid peroxidation and a deteriorated endogenous antioxidant defense system (glutathione, catalase and superoxide dismutase). However, all the above-mentioned obesity-related complications were dose-dependently and significantly inhibited after 84 days of thye consecutive oral administration of 125, 250 and 500 mg/kg YPh. In addition, YPh dose-dependently depleted the liver endogenous antioxidant defense system and inhibited hepatic lipid peroxidation. Overall, the effects of 250 mg/kg YPh on HFD-induced obesity and related complications were similar or more potent than those of 10 mg/kg simvastatin. These results indicate that YPh is a promising new potent medicinal ingredient for possible use in the treatment of obesity and related complications.

Abstract 5099: Development of novel antimetabolite, DFP-11207, with self-toxicity protection and its promising preclinical and clinical profiles

Abstract Background: For over 50 years, 5-fluorouracil (5-FU) has played a critical role in the systemic chemotherapy of various gastrointestinal cancers including gastric, colorectal and pancreatic cancer which are the leading causes of cancer death globally. Although systemic cytotoxics such as 5-FU can modestly prolong overall survival in the adjuvant and advanced settings, but at the cost of unpleasant toxicities. Therefore better drug formulations are desirable. Methods: To this end, we developed a new conceptual fluoropyrimidine, DFP-11207 that is engineered to reduce toxicity without loss of antitumor activity as well in addition to providing sustained concentrations of the active anticancer moiety. DFP-11207 contains three components in one formulation: 1-ethoxymethyl-5-fluorouracil (EM-FU) as a prodrug of 5-FU, 5-chloro-2,4-dihyroxypyridine (CDHP) as a potent inhibitor of 5-FU degradation and cytrazinic acid (CTA) as a gastrointestinal regulator of 5-FU phosphorylation. Results: In in vitro metabolism studies using cell-free extracts from plasma, liver and tumors or intact tumor cells, DFP-11207 was rapidly hydrolyzed to 3 components and subsequently EM-FU was specifically converted to 5-FU by liver microsomes, and CDHP and CTA strongly inhibited 5-FU degradation and phosphorylation, respectively. Following consecutive oral administration to human tumor-bearing nude rats, DFP-11207 attained favorable antitumor efficacy and long-sustained PK profiles with lack of GI- and myelo-toxicities. Next, in investigational clinical study in patients with solid tumors, 12 patients were treated at 8 unique dose levels of DFP-11207, ranging from 40 to 440mg/m2/day by each 1 pt. MTD and RD of daily and 28-day consecutive DFP-11207 was found to be 440 (n=2) and 330 mg/m2 (n=6), respectively. The main AEs were nausea,, anemia, neutropenia, febrile neutropenia but these events were very mild, and no thrombocytopenia was observed as expected. Furthermore, review of the preliminary PK data, DFP-11207 at 330 mg/m2 resulted in a desirable low but efficacious (~15- 30 ng/ml) steady-state plasma concentration of 5-FU. Interestingly, some of patients heavily treated with therapeutic drugs had a stable-disease for long periods. Conclusion: Our preclinical and early clinical data with DFP-11207 suggest that it’s a promising compound for the treatment of gastrointestinal cancers and can overcome the shortcomings of all other oral fluoropyrimidines. Citation Format: Masakazu Fukushima, Kenzo Iizuka, Kokoro Eshima, Chung Zhang, Cheng Jin, Kiyoshi Eshima, Jaffer Ajani. Development of novel antimetabolite, DFP-11207, with self-toxicity protection and its promising preclinical and clinical profiles [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5099. doi:10.1158/1538-7445.AM2017-5099

Montelukast ameliorates streptozotocin-induced cognitive impairment and neurotoxicity in mice

Extensive studies have demonstrated that neuroinflammation is associated with Alzheimer’s disease (AD) and cysteinyl leukotriene receptor 1 (CysLT1R) was involved in neuroinflammation. Montelukast, a highly selective CysLT1R antagonist, has been reported to attenuate learning and memory impairments in the amyloid-β-induced mouse model of AD. However, whether montelukast also exerts beneficial effects on streptozotocin (STZ)-induced memory deficits in mice is not well known. In the present study, we aimed to investigate the effects of montelukast on STZ-induced cognitive impairment, neuroinflammation and apoptosis in mice. Our data showed that intra-hippocampal microinfusion of STZ resulted in learning and memory impairments, including increased escape latency during acquisition trials and decreased exploratory activities in the probe trial in Morris watermaze (MWM) task, and decreased number of correct choices and increased latency to enter the shock-free compartment in Y-maze test, and caused neuroinflammatory and apoptotic responses, evidenced by increments of nuclear NF-κB p65, TNF-α, IL-1β, cleaved caspase-3, Bax as well as decreased expression of Bcl-2 in hippocampus. Interestingly, STZ treatment led to up-regulation of protein and mRNA of CysLT1R in hippocampus. Of note, consecutive oral administration of montelukast (1 or 2mg/kg, 3 weeks) remarkably attenuated these effects induced by STZ. However, montelukast had no effect on normal mice. These results suggest that montelukast improves memory impairment and inhibits neuroinflammation and apoptosis in mice exposed to STZ. Montelukast may provide a novel strategy for treating or preventing AD.

Abstract PR277

Background & Objectives: Newborns are often sedated during MRI but sedation itself creates adverse events and the management is more challenging in the MR environment (1). We describe our initial experience with oral 30% glucose administration through a teat during MRI for imaging of newborns. Materials & Methods: Nineteen newborns were assessed retrospectively after institutional research and ethics committee approval. 0.5-1 mL 30% glucose solution was administered orally. The motionless and sleepiness of newborns were evaluated after suckling. If the target conditions was not achieved, 0.5-1 mL increments of glucose was added. After 2 consecutive oral glucose administration the newborns who did not keep motionless or asleep and had motion artefacts were sedated with midazolam. The routine blood glucose level measurement of the newborns after imaging procedures in the ICU was performed for each patient. Results: Fifteen of 19 newborns quietened, kept motionless and slept during the procedure with administration of oral 30% glucose solution. However, 4 of 19 newborns did not keep motionless or asleep after 2 additional dosages of 30% glucose for induction and were needed further sedation. Conclusion: Majority of newborns completed MRI examination with oral 30% glucose administration without a need for sedation. Thereby, we recommend and support the integration of this safe and reliable technique into routine practice for newborns during MRI.

The Extract of Roots of Sophora flavescens Enhances the Recovery of Motor Function by Axonal Growth in Mice with a Spinal Cord Injury

Although axonal extension to reconstruct spinal tracts should be effective for restoring function after spinal cord injury (SCI), chondroitin sulfate proteoglycan (CSPG) levels increase at spinal cord lesion sites, and inhibit axonal regrowth. In this study, we found that the water extract of roots of Sophora flavescens extended the axons of mouse cortical neurons, even on a CSPG-coated surface. Consecutive oral administrations of S. flavescens extract to SCI mice for 31 days increased the density of 5-HT-positive axons at the lesion site and improved the motor function. Further, the active constituents in the S. flavescens extract were identified. The water and alkaloid fractions of the S. flavescens extract each exhibited axonal extension activity in vitro. LC/MS analysis revealed that these fractions mainly contain matrine and/or oxymatrine, which are well-known major compounds in S. flavescens. Matrine and oxymatrine promoted axonal extension on the CSPG-coated surface. This study is the first to demonstrate that S. flavescens extract, matrine, and oxymatrine enhance axonal growth in vitro, even on a CSPG-coated surface, and that S. flavescens extract improves motor function and increases axonal density in SCI mice.

흰 민들레 뿌리의 항염증 및 산화 스트레스 개선 효과

한방에서 염증 치료 목적으로 사용하는 흰 민들레 뿌리의 열수 추출물을 제조하여 항염증 효과 및 산화 스트레스 개선효과를 확인하고자 하였다. ICR mice에 lipopolysaccharide (LPS)를 1회 복강 투여하여(10 mg/kg bw) 염증 및 산화스트레스를 유발하고, 한국에만 자생하는 흰 민들레 뿌리열수 추출물을 1.4 g/kg bw 농도로 2회 경구 투여하였다. LPS에 의한 염증 유발은 체중 감소 현상으로 확인하였고, 민들레 뿌리 열수 추출물 투여군은 유의적으로 체중이 증가하여 염증 억제 효과가 관찰되었다. LPS 처리는 혈액의 중성지방질, 혈액 및 간의 reactive oxygen species(ROS), peroxynitrite 농도 그리고 간의 지질과산화 수준을 증가시켰다(P0.05). LPS를 처리한 대조군에 비해 흰 민들레 뿌리열수 추출물 투여군은 혈중 및 간의 중성지방질, ROS, peroxynitrite 농도 그리고 간의 지질과산화물 수준이 유의적으로 감소하였다(P0.05). 이는 흰 민들레 뿌리 열수 추출물이 간의 NF-κB 전사인자 및 이의 조절을 받는 COX-2의 단백질 발현을 억제함으로써 염증 관련 매개체 및 유리기 생성을 유의적으로 억제하였기 때문으로 생각된다. LPS 처리군에서 간의 중성지방질 농도가 감소한 것은 염증에 의해 간 조직이 손상되어 중성지방질의 합성이 억제되었기 때문으로 생각된다. 본 연구는 한방에서 사용하여 온 한국 토종 흰 민들레 뿌리가 NF-κB 전사인자의 발현을 억제함으로써 염증 및 산화 스트레스를 개선하는 효과가 있음을 확인하였다.

Toxicity studies of coumarin 6-encapsulated polystyrene nanospheres conjugated with peanut agglutinin and poly(N-vinylacetamide) as a colonoscopic imaging agent in rats

We are investigating an imaging agent that detects early-stage primary colorectal cancer on the mucosal surface in real time under colonoscopic observation. The imaging agent, which is named the nanobeacon, is fluorescent nanospheres conjugated with peanut agglutinin and poly(N-vinylacetamide). Its potential use as an imaging tool for colorectal cancer has been thoroughly validated in numerous studies. Here, toxicities of the nanobeacon were assessed in rats. The nanobeacon was prepared according to the synthetic manner which is being established as the Good Manufacturing Practice-guided production. The rat study was performed in accordance with Good Laboratory Practice regulations. No nanobeacon treatment-related toxicity was observed. The no observable adverse effect levels (NOAEL) of the nanobeacon in 7-day consecutive oral administration and single intrarectal administration were estimated to be more than 1000mg/kg/day and 50mg/kg/day, respectively. We concluded that the nanobeacon could be developed as a safe diagnostic agent for colonoscopy applications. From the Clinical EditorColon cancer remains a major cause of death. Early detection can result in early treatment and thus survival. In this article, the authors tested potential systemic toxicity of coumarin 6-encapsulated polystyrene nanospheres conjugated with peanut agglutinin (PNA) and poly(N-vinylacetamide) (PNVA), which had been shown to bind specifically to colonic cancer cells and thus very promising in colonoscopic detection of cancer cells.

Consecutive oral administration of Bifidobacterium longum MM-2 improves the defense system against influenza virus infection by enhancing natural killer cell activity in a murine model.

Bifidobacterium, one of the major components of intestinal microflora, shows anti-influenza virus (IFV) potential as a probiotic, partly through enhancement of innate immunity by modulation of the intestinal immune system. Bifidobacterium longum MM-2 (MM-2), a very safe bacterium in humans, was isolated from healthy humans and its protective effect against IFV infection in a murine model shown. In mice that were intranasally inoculated with IFV, oral administration of MM-2 for 17 consecutive days improved clinical symptoms, reduced mortality, suppressed inflammation in the lower respiratory tract, and decreased virus titers, cell death, and pro-inflammatory cytokines such as IL-6 and TNF-α in bronchoalveolar lavage fluid. The anti-IFV mechanism of MM-2 involves innate immunity through significant increases in NK cell activities in the lungs and spleen and a significant increase in pulmonary gene expression of NK cell activators such as IFN-γ, IL-2, IL-12 and IL-18. Even in non-infected mice, MM-2 administration also induced significant enhancement of both IFN-γ production by Peyer's patch cells (PPs) and splenetic NK cell activity. Oral administration of MM-2 for 17 days activates systemic immunoreactivity in PPs, which contributes to innate immunity, including NK cell activation, resulting in an anti-IFV effect. MM-2 as a probiotic may function as a prophylactic agent in the management of an IFV epidemic.

Safety of the cell-penetrating peptide penetratin as an oral absorption enhancer.

Novel delivery technology using cell-penetrating peptides (CPPs) have recently shown their potential and are emerging as promising candidates for an oral protein and peptide delivery systems. As with for the development of any absorption enhancer that is meant to function across an epithelial layer covering a surface highly exposed to pathogens such as the intestines, concern arises about the safety of such enhancers. The purpose of this study was to investigate the effect of 7 d of consecutive oral administrations of CPPs and a typical enterotoxin, lipopolysaccharide (LPS) to mice to determine the degree, if any, of damage caused to the hepatic tissue. Following the 7-d dosing regimen, we could not detect significantly increased levels of the liver enzymes alanine aminotransferase and aspartate aminotransferase in plasma of mice treated with CPP and LPS compared to the controls, whereas heightened levels were observed in animals receiving the bile salt. In conclusion, the repeated use of CPPs as an oral absorption enhancer for macromolecules was found to be a safe strategy.

Study on impact of vinegar processing on toxic effect of Stellera chamaejasme

To study the scientific theory on detoxification (attenuation) of Stellera chamaejasme (ScL) by processing and the impact on drug effect of ScL before and after being processed with vinegar. The difference in ingredients of ScL before and after being processed vinegar was compared by using PHLC-MS technique. A subcutaneously transplanted tumor model of H22 hepatoma was established to compare the lethal effect and weight change between tumor-loaded mice and normal mice. After consecutive oral administration in tumor-loaded mice, the impacts on tumors and immune organs were compared before and after being processed with vinegar. Luciferase Report Gene was employed to investigate the target genes TGF-beta, AP1 and NF-kappaB. The LD50 (median fatal dose) of extract Zp1102 exhibited higher than that of the processed one Zp1103, that is 9. 89 g x kg(-1) vs. 16.85 g x kg(-1). According to the test, Zp1102 showed more effective anti-tumor activities in vivo than that of Zp1103 in a same dosage, with the tumor inhibitory rate 36.24% (P < 0.01) at the dosage of2 g x kg(-1) and 34.40% (P < 0.05) at the dosage of 1 g x kg(-1). At the dosage of 1 g x kg(-1), Zp1102 showed a tumor inhibitory rate of 34.52% (P < 0.05), much higher from 21.55% in Zp1103. Both Zp1102 and Zp1103 had basically no impact on the report gene NF-kappaB, besides that Zp1102 up-regulated the report gene after increase in NF-kappaB concentration and down-regulated TGF-beta, but Zp1103 can only up-regulate NF-kappaB expression without any impact on TGF-beta. Processed ScL extracts show less toxic than unprocessed extracts and slight reduction in anti-tumor activity, which may be related to the regulation of transforming growth factor TGF-beta.