Investigation of the Factors Responsible for the Poor Oral Bioavailability of Acacetin in Rats: Physicochemical and Biopharmaceutical Aspects.

Acacetin, an important ingredient of acacia honey and a component of several medicinal plants, exhibits therapeutic effects such as antioxidative, anticancer, anti-inflammatory, and anti-plasmodial activities. However, to date, studies reporting a systematic investigation of the in vivo fate of orally administered acacetin are limited. Moreover, the in vitro physicochemical and biopharmaceutical properties of acacetin in the gastrointestinal (GI) tract and their pharmacokinetic impacts remain unclear. Therefore, in this study, we aimed to systematically investigate the oral absorption and disposition of acacetin using relevant rat models. Acacetin exhibited poor solubility (≤119 ng/mL) and relatively low stability (27.5-62.0% remaining after 24 h) in pH 7 phosphate buffer and simulated GI fluids. A major portion (97.1%) of the initially injected acacetin dose remained unabsorbed in the jejunal segments, and the oral bioavailability of acacetin was very low at 2.34%. The systemic metabolism of acacetin occurred ubiquitously in various tissues (particularly in the liver, where it occurred most extensively), resulting in very high total plasma clearance of 199 ± 36 mL/min/kg. Collectively, the poor oral bioavailability of acacetin could be attributed mainly to its poor solubility and low GI luminal stability.

Butyrate enhances the efficacy of radiotherapy via FOXO3A in colorectal cancer patient­derived organoids.

Enhancing the radioresponsiveness of colorectal cancer (CRC) is essential for local control and prognosis. However, consequent damage to surrounding healthy cells can lead to treatment failure. We hypothesized that short­chain fatty acids (SCFAs) could act as radiosensitizers for cancer cells, allowing the administration of a lower and safer dose of radiation. To test this hypothesis, the responses of three­dimensional­cultured organoids, derived from CRC patients, to radiotherapy, as well as the effects of combined radiotherapy with the SCFAs butyrate, propionate and acetate as candidate radiosensitizers, were evaluated via reverse transcription­quantitative polymerase chain reaction, immunohistochemistry and organoid viability assay. Of the three SCFAs tested, only butyrate suppressed the proliferation of the organoids. Moreover, butyrate significantly enhanced radiation­induced cell death and enhanced treatment effects compared with administration of radiation alone. The radiation­butyrate combination reduced the proportion of Ki­67 (proliferation marker)­positive cells and decreased the number of S phase cells via FOXO3A. Meanwhile, 3/8 CRC organoids were found to be non­responsive to butyrate with lower expression levels of FOXO3A compared with the responsive cases. Notably, butyrate did not increase radiation­induced cell death and improved regeneration capacity after irradiation in normal organoids. These results suggest that butyrate could enhance the efficacy of radiotherapy while protecting the normal mucosa, thus highlighting a potential strategy for minimizing the associated toxicity of radiotherapy.

Enhancing Butyrate Production, Ruminal Fermentation and Microbial Population through Supplementation with Clostridium saccharobutylicum.

Butyrate is known to play a significant role in energy metabolism and regulating genomic activities that influence rumen nutrition utilization and function. Thus, this study investigated the effects of an isolated butyrate-producing bacteria, Clostridium saccharobutylicum, in rumen butyrate production, fermentation parameters and microbial population in Holstein-Friesian cow. An isolated butyrate-producing bacterium from the ruminal fluid of a Holstein-Friesian cow was identified and characterized as Clostridium saccharobutylicum RNAL841125 using 16S rRNA gene sequencing and phylogenetic analyses. The bacterium was evaluated on its effects as supplement on in vitro rumen fermentation and microbial population. Supplementation with 106 CFU/ml Clostridium saccharobutylicum increased (p < 0.05) microbial crude protein, butyrate and total volatile fatty acids concentration but had no significant effect on NH3-N at 24 h incubation. Butyrate and total VFA concentrations were higher (p < 0.05) in supplementation with 106 CFU/ml Clostridium saccharobutylicum compared with control, with no differences observed for total gas production, NH3-N and propionate concentration. However, as the inclusion rate (CFU/ml) of C. saccharobutylicum was increased, reduction of rumen fermentation values was observed. Furthermore, butyrate-producing bacteria and Fibrobacter succinogenes population in the rumen increased in response with supplementation of C. saccharobutylicum, while no differences in the population in total bacteria, protozoa and fungi were observed among treatments. Overall, our study suggests that supplementation with 106 CFU/ml C. saccharobutylicum has the potential to improve ruminal fermentation through increased concentrations of butyrate and total volatile fatty acid, and enhanced population of butyrate-producing bacteria and cellulolytic bacteria F. succinogenes.

Modulation of Rat Hepatic CYP1A and 2C Activity by Honokiol and Magnolol: Differential Effects on Phenacetin and Diclofenac Pharmacokinetics In Vivo.

Honokiol (2-(4-hydroxy-3-prop-2-enyl-phenyl)-4-prop-2-enyl-phenol) and magnolol (4-Allyl-2-(5-allyl-2-hydroxy-phenyl)phenol) are the major active polyphenol constituents of Magnolia officinalis (Magnoliaceae) bark, which has been widely used in traditional Chinese medicine (Houpu Tang) for the treatment of various diseases, including anxiety, stress, gastrointestinal disorders, infection, and asthma. The aim of this study was to investigate the direct effects of honokiol and magnolol on hepatic CYP1A and 2C-mediated metabolism in vitro using rat liver microsomes and in vivo using the Sprague-Dawley rat model. Honokiol and magnolol inhibited in vitro CYP1A activity (probe substrate: phenacetin) more potently than CYP2C activity (probe substrate: diclofenac): The mean IC50 values of honokiol for the metabolism of phenacetin and diclofenac were 8.59 µM and 44.7 µM, while those of magnolol were 19.0 µM and 47.3 µM, respectively. Notably, the systemic exposure (AUC and Cmax) of phenacetin, but not of diclofenac, was markedly enhanced by the concurrent administration of intravenous honokiol or magnolol. The differential effects of the two phytochemicals on phenacetin and diclofenac in vivo pharmacokinetics could at least be partly attributed to their lower IC50 values for the inhibition of phenacetin metabolism than for diclofenac metabolism. In addition, the systemic exposure, CL, and Vss of honokiol and magnolol tended to be similar between the rat groups receiving phenacetin and diclofenac. These findings improve our understanding of CYP-mediated drug interactions with M. officinalis and its active constituents.

Effects of supplementing Barodon, Bacillus subtilis, and Ampbio on growth performance, biochemical metabolites, and hormone levels in Korean native heifers.

An experiment was conducted to evaluate the effects of supplementing feed additives of Barodon®, effective microorganism® (Bacillus (B.) subtilis), and Ampbio® on the growth performance, blood metabolites, stress, and reproductive hormone levels of Korean native heifers. A total of 48 Korean native heifers were assigned to four groups with 6 heifers in each group. The groups were control, Barodon (0.2%), beneficial microorganism (1%), and Ampbio (1%). Animals in all groups were fed a basal diet composed of selected feed additives and water ad libitum for 99 days. Results showed that there are significant changes in body weight and daily gain in the Ampbio-supplemented group as compared with the control and other feed additives groups (p < 0.05). The increased level of feed intake (7.30 ± 0.03 kg) and feed requirements (10.81 ± 0.52 kg) was observed in the Ampbio-fed group followed by the effective microorganism (EM), Barodon, and control groups. There were no significant changes in albumin, glucose, glutamic oxaloacetic transaminase (SGOT), serum pyruvic transaminase (SGPT), and total protein level, but the decreased levels of total cholesterol and triglycerides and the increased level of blood urea nitrogen were noted in the Ampbio-fed group as compared with the control and other feed additive groups. The reduced level of cortisol (p < 0.05) and elevated levels of progesterone and estradiol (p > 0.05) were noted in the Ampbio-fed group as compared to the other feed additive groups. It is therefore concluded that incorporation of Barodon, EM (B. subtilis), and Ampbio in the recommended diet improved the growth and health performance of Korean native heifers.

Changes in microbial growth, carotenoids, and water-soluble tannin content of ripe persimmon beverage after ultra-high pressure treatment.

To avoid the loss of carotenoids and increasing the tannin content associated with pasteurization, we tested ultra-high pressure treatment of ripe persimmon beverage. We compared microbial counts (aerobic bacteria, coliforms, and mould), carotenoid contents, and water-soluble tannin contents between heat- and ultra-high pressure-treated beverages. No microbial contamination was detected after pasteurization or ultra-high pressure treatment at 400 MPa for more than 5 min. Ultra-high pressure treatment significantly prevented the reduction in carotenoids (lutein, zeaxanthin, ß-cryptoxanthin, ß-carotene, lycopene), with losses of 3.9-28.7%, as compared to the 65% loss after pasteurization. Moreover, ultra-high pressure did not induce an increase in water-soluble tannin, which causes astringent taste, whereas water-soluble tannins were increased three times by heat treatment. In conclusion, ultra-high pressure showed the same microbial control effect as pasteurization, while it did not cause carotenoid degeneration and increased tannin and thus, it better maintained the quality of ripe persimmon beverage.

Benefits of stereotactic ablative radiotherapy combined with incomplete transcatheter arterial chemoembolization in hepatocellular carcinoma.

BACKGROUND: This study aimed to evaluate the effect of stereotactic ablative radiotherapy (SABR) after incomplete transcatheter arterial chemoembolization (TACE) in hepatocellular carcinoma (HCC) patients. METHODS: The study enrolled 178 HCC patients initially treated with TACE between 2006 and 2011. Patients were included if they had Barcelona Clinic Liver Cancer stage 0 or A, ≤3 nodules with a total sum of longest diameter ≤10 cm, Child-Turcotte-Pugh score of ≤7, no major vessel invasion, and no extra-hepatic metastases. RESULTS: Twenty-four patients achieved a complete response to TACE (group 1). Among those with incomplete response, 47 patients received other curative treatments (group 2), 37 received SABR (group 3), and 70 received non-curative treatments (group 4). The 2-year overall survival (OS) rates for groups 1, 2, 3, and 4 were 88 %, 81 %, 73 %, and 54 %, respectively. The corresponding 5-year OS rates were 50 %, 58 %, 53 %, and 28 %, respectively. CONCLUSIONS: Patients treated with SABR after incomplete TACE had similar survival outcomes to those achieving complete response to TACE or receiving curative treatments. However, patients receiving non-curative treatments had significantly lower survival rates than the other groups. Therefore, if SABR was indicated at the initial diagnosis, it might be recommended after TACE failure.

High body clearance and low oral bioavailability of alantolactone, isolated from Inula helenium, in rats: extensive hepatic metabolism and low stability in gastrointestinal fluids.

Alantolactone (ALA) is a major bioactive sesquiterpene lactone present in the roots of Inula helenium L. (Asteraceae) which has been used widely in traditional medicine against various diseases such as asthma, cancer and tuberculosis. The pharmacologic activities of alantolactone have been well characterized, yet information on the physicochemical and pharmacokinetic properties of alantolactone and their mechanistic elucidation are still limited. Thus, this study aims to investigate the oral absorption and disposition of alantolactone and their relevant mechanisms. Log P values of alantolactone ranged from 1.52 to 1.84, and alantolactone was unstable in biological samples such as plasma, urine, bile, rat liver microsomes (RLM) and simulated gastrointestinal fluids. The metabolic rate of alantolactone was markedly higher in rat liver homogenates than in the other tissue homogenates. A saturable and concentration-dependent metabolic rate profile of alantolactone was observed in RLM, and rat cytochrome P450 (CYP) 1 A, 2C, 2D and 3 A subfamilies were significantly involved in its hepatic metabolism. Based on the well-stirred model, the hepatic extraction ratio (HER) was estimated to be 0.890-0.933, classifying alantolactone as a drug with high HER. Moreover, high total body clearance (111 ± 41 ml/min/kg) and low oral bioavailability (0.323%) of alantolactone were observed in rats. Taken together, the present study demonstrates that the extensive hepatic metabolism, at least partially mediated by CYP, is primarily responsible for the high total body clearance of alantolactone, and that the low oral bioavailability of alantolactone could be attributed to its low stability in gastrointestinal fluids and a hepatic first-pass effect in rats. Copyright © 2016 John Wiley & Sons, Ltd.

Modulation of Cytochrome P450 Activity by 18ß-Glycyrrhetic Acid and its Consequence on Buspirone Pharmacokinetics in Rats.

The aim of this study was to elucidate the inhibition mechanism of 18ß-glycyrrhetic acid (GLY) on cytochrome P450 (CYP) activity and in vivo pharmacokinetic consequences of single GLY dose in rats. An in vitro CYP inhibition study in rat liver microsomes (RLM) was conducted using probe substrates for CYPs. Then, an in vivo pharmacokinetics of intravenous and oral buspirone (BUS), a probe substrate for CYP3A, was studied with the concurrent administration of oral GLY in rats. In the in vitro CYP inhibition study, CYP3A was involved in the metabolism of GLY. Moreover, GLY inhibited CYP3A activity with an IC50 of 20.1 ± 10.7 µM via a mixed inhibition mechanism. In the in vivo rat pharmacokinetic study, single oral GLY dose enhanced the area under plasma concentration-time curve (AUC) of intravenous and oral BUS, but the extent of increase in AUC was only minimal (1.12-1.45 fold). These results indicate that GLY can inhibit the in vitro CYP3A-mediated drug metabolism in RLM via a mixed inhibition mechanism. However, the impact of single oral GLY dose on the pharmacokinetics of BUS in rats was limited, showing that GLY could function as merely a weak inhibitor for CYP3A-mediated drug metabolism in vivo. Copyright © 2015 John Wiley & Sons, Ltd.

In vitro and in vivo evaluation of the effect of puerarin on hepatic cytochrome p450-mediated drug metabolism.

Puerarin (8-ß-D-glucopyranosyl-7-hydroxy-3-(4-hydroxyphenyl)-4H-1-benzopyran-4-one) is a major pharmacological component of Puerariae Radix, the root of Pueraria lobata. We investigated the effect of puerarin on hepatic cytochrome P450-mediated drug metabolism in rats and humans. The in vitro cytochrome P450 inhibitory effect of puerarin in human and rat liver microsomes was evaluated using the following model cytochrome P450 substrates: phenacetin for CYP1A, diclofenac for CYP2C, dextromethorphan for CYP2D, and testosterone for CYP3A. The in vivo pharmacokinetics of intravenous and oral buspirone, a probe substrate for CYP3A, was studied with single simultaneous intravenous coadministration of puerarin in rats. In the in vitro cytochrome P450 inhibition study, the rate of disappearance of testosterone was significantly reduced in the presence of 10 µM PU, while that of other cytochrome P450 substrates was not significantly affected in both human and rat liver microsomes, suggesting that puerarin inhibits the in vitro hepatic CYP3A-mediated metabolism in the human and rat systems (IC50 = 15.5 ± 3.9 µM). After intravenous administration of buspirone with single simultaneous coadministration of intravenous puerarin at a dose of 10 mg/kg in rats, the total area under the plasma concentration-time curve from time zero to time infinity was increased while time-averaged total body clearance decreased. When buspirone was orally administered in rats with the 10 mg/kg intravenous puerarin coadministration, both total area under the plasma concentration-time curve from time zero to time infinity and the extent of absolute oral bioavailability were significantly increased. Therefore, results of the in vitro microsomal and in vivo pharmacokinetic studies suggest the possible inhibition of hepatic CYP3A-mediated drug metabolism by puerarin administration, potentially leading to metabolism-mediated herb-drug interactions with clinical significance.

Effects of fermented persimmon extract supplements on chemical composition and fermentation characteristics of barley silage.

This study was conducted to estimate the effect of fermented persimmon extract (FPE) supplementation levels on chemical composition and fermentation characteristics of barley silage. Barley forage was harvested at 25% dry matter and chopped to 5 cm lengths. The FPE was applied at 0, 0.4, 0.8 and 1.6 g/kg of barley forage (fresh weight basis). After sub-sampling of barley forages (0 day), a total of 64 laboratory mini silos (5 kg capacity) were ensiled to generate quadruplicate data from each of four treatments for four ensiling durations (2, 6, 40 and 120 days). The chemical compositions and in vitro digestibility of barley forage (0 day) were not affected by FPE supplementation. However, the content (L, P = 0.035) and in vitro digestibility (L, P = 0.041) of neutral detergent fiber on 120-day barley silage decreased by increasing FPE supplements. Acetate content of barley silage ensiled for 120 days increased (Q, P = 0.004) by increasing FPE supplements, whereas pH (L, P < 0.001), lactate content (Q, P < 0.001) and lactate-to-acetate ratio (Q, P < 0.001) decreased. These results indicate that addition of FPE can increase the acetate content which has potential to increase aerobic stability.

Effect of bisphosphonates on bone mineral density and fracture prevention in gastric cancer patients after gastrectomy.

GOAL: To evaluate the effectiveness of bisphosphonates in preventing fractures in gastric cancer patients by increasing bone mineral density (BMD). BACKGROUND: The effectiveness of bisphosphonates is questionable in gastric cancer patients who have undergone gastrectomy, although they display a high prevalence of osteoporosis. STUDY: Forty-seven gastric cancer patients with osteoporosis were retrospectively analyzed. All patients were supplemented with calcium and vitamin D. Twenty-four patients were treated with bisphosphonate (bisphosphonate group) and 23 patients were untreated (control group). Fractures, severe bone pain, and adverse effects of bisphosphonates were monitored. BMD of the lumbar spine and femoral neck were measured before and 1-year treatment with bisphosphonates by dual-energy X-ray absorptiometry. RESULTS: During a 1-year follow-up, 7 of the 47 (15%) patients developed new fractures. The bisphosphonate-treated group had a significantly lower fracture rate than the control group (n=1 vs. 6, P<0.05). Lumbar spine BMD increased in both groups (0.047 ± 0.03 vs. 0.021 ± 0.03 g/cm², respectively), whereas femoral neck BMD increased only in the bisphosphonate group (0.032 ± 0.03 vs. -0.004 ± 0.02 g/cm², respectively). Furthermore, the bisphosphonate group showed greater increases in lumbar spine and femoral neck BMDs than the controls (P<0.05). No difference was found between alendronate and risedronate in terms of BMD at follow-up. CONCLUSIONS: Therapy using bisphosphonates might be effective at increasing BMD and reducing fracture risk in gastric cancer patients after gastrectomy. Further well-designed randomized controlled trials are needed for confirmation.

Chronic stroke longitudinal motor improvements: cumulative learning evidence found in the upper extremity.

BACKGROUND: The purpose was to determine the cumulative longitudinal effects of upper extremity distributed practice with variable treatment protocols involving EMG-triggered neuromuscular stimulation and coupled bilateral movements. METHODS: Sixteen chronic stroke subjects were randomly selected to complete 5 effective upper extremity treatment protocols over 12 months. The subjects were randomly assigned to 1 of 2 treatment orders. Consistent across the orders and protocols, the participants completed 90 min of training per day for 4 days during separate 2-week rehabilitation periods. RESULTS: Data for the 5 primary outcome measures were analyzed in separate mixed design ANOVAs (treatment order x test session: 2 x 6). The analyses revealed distinct cumulative treatment evidence later in training in comparison to the baseline motor capabilities: (1) higher number of blocks moved; (2) higher percentage of blocks moved by the impaired hand; (3) faster motor reaction time (peripheral component), and (4) faster total reaction time. CONCLUSIONS: These chronic stroke patients displayed robust cumulative motor improvement effects from the longitudinally distributed practice of active neuromuscular stimulation and coupled bilateral movements.

Coupled bilateral movements and active neuromuscular stimulation: intralimb transfer evidence during bimanual aiming.

Motor improvements in chronic stroke recovery accrue from coupled protocols of bilateral movements and active neuromuscular stimulation. This experiment investigated coupled protocols and within-limb transfer between distal and proximal joint combinations. The leading question focused on within-limb transfer of coupled protocols on distal joints to a bimanual aiming task that involved proximal joints. Twenty-six volunteers completed one of three motor recovery protocols according to group assignments: (1) coupled bilateral involved concurrent wrist/finger movements on the unimpaired limb coupled with active stimulation on the impaired limb; (2) unilateral/active stimulation involved neuromuscular electromyogram-triggered stimulation on the impaired wrist/fingers; and (3) no protocol (control group). During the pretest and posttest, subjects performed transverse plane target aiming movements (29 cm) with vision available. The coupled bilateral group showed positive intralimb transfer post-treatment when both arms moved simultaneously. During the posttest, the coupled bilateral group displayed improved movement time, higher peak limb velocity, less variability in peak velocity, and less percentage of total movement time in the deceleration phase than during the pretest. The evidence confirms that within-limb transfer from distal joint training to proximal joint combinations is viable and generalizable in chronic stroke rehabilitation. Moreover, these intralimb transfer findings extend the evidence favoring motor improvements for coupled bilateral protocols during chronic stroke.

Chronic stroke motor recovery: duration of active neuromuscular stimulation.

Active neuromuscular stimulation is an effective behavioral intervention for motor recovery improvements after a stroke. However, the most effective active neuromuscular stimulation durations have not been determined. The present experiment investigated active stimulation durations (0, 5, and 10 s) coupled with bilateral movements on progress toward motor recovery in wrist and finger extension. Twenty-six stroke survivors with chronic hemiparesis were randomly assigned to the stimulation duration groups, and subjects completed 4 days of rehabilitation training over a 2-week period. Mixed design analyses of variance on the Box and Block scores, chronometric reaction times, and force modulation of the sustained muscle contraction task revealed distinct motor recovery improvements for both the 5 and 10 s stimulation duration groups in comparison to the 0 s duration control group. Further, the number of blocks moved by the 10-s duration group exceeded those moved by the 5-s duration group. In conclusion, the 5 and 10 s duration active stimulation/bilateral movement groups decreased residual motor dysfunctions that persisted beyond 12 months post stroke.