Cell-free culture supernatant of Lactobacillus curvatus Wikim38 inhibits RANKL-induced osteoclast differentiation and ameliorates bone loss in ovariectomized mice.

This study was conducted to investigate the inhibitory effects of the cell-free culture supernatant of Lactobacillus curvatus Wikim 38 (LC38-CS) on RANKL-induced osteoclast differentiation and bone loss in a mice model of ovariectomy-induced post-menopausal osteoporosis. LC38-CS inhibited the RANKL-induced differentiation of bone marrow-derived macrophages (BMDMs) into osteoclasts in a dose-dependent manner. F-actin ring formation and bone resorption were also reduced by LC38-CS treatment of RANKL-treated BMDMs. In addition, LC38-CS decreased the RANKL-induced activation of the TRAF6/NF-κB/MAPKs axis at the early stage and the expression of osteoclastogenesis-related genes in BMDMs treated with RANKL. PRMT1 and ADMA levels, new biomarkers for osteoclastogenesis, were decreased by LC38-CS treatment. The administration of LC38-CS increased bone volume and bone mineral density in ovariectomized mice in µ-CT analysis. These findings suggest that LC38-CS inhibited RANKL-induced osteoclast differentiation by the downregulation of molecular mechanisms and exerted anti-osteoporotic effects.

A phase III study comparing SB3 (a proposed trastuzumab biosimilar) and trastuzumab reference product in HER2-positive early breast cancer treated with neoadjuvant-adjuvant treatment: Final safety, immunogenicity and survival results.

BACKGROUND: The equivalent efficacy between SB3, a proposed trastuzumab biosimilar, and the trastuzumab reference product (TRZ) in terms of the breast pathologic complete response rate after neoadjuvant therapy in patients with early or locally advanced human epidermal growth factor receptor 2-positive breast cancer was demonstrated in the previous report. Here, we report the final safety, immunogenicity and survival results after neoadjuvant-adjuvant treatment. PATIENTS AND METHODS: Patients were randomised 1:1 to receive neoadjuvant SB3 or TRZ for 8 cycles concurrently with chemotherapy (4 cycles of docetaxel followed by 4 cycles of 5-fluorouracil/epirubicin/cyclophosphamide). Patients then underwent surgery, followed by 10 cycles of adjuvant SB3 or TRZ as randomised. End-points included safety, immunogenicity, event-free survival (EFS) and overall survival through the adjuvant period. RESULTS: Of 875 patients randomised, 764 (SB3, n = 380; TRZ, n = 384) completed the study. The median follow-up duration was 437 days in the SB3 group and 438 days in the TRZ group. The incidence of treatment-emergent adverse events was comparable between groups (SB3, 97.5%; TRZ, 96.1%) during the overall study period. Up to the end of study, the overall incidence of antidrug antibody was low in both treatment groups (3 patients each). EFS was comparable between groups with a hazard ratio (SB3/TRZ) of 0.94 (95% confidence interval, 0.59-1.51) and EFS rates at 12 months of 93.7% for SB3 and 93.4% for TRZ. CONCLUSIONS: Final safety, immunogenicity and survival results of this study further support the biosimilarity established between SB3 and TRZ. TRIAL REGISTRATION: ClinicalTrials.gov (NCT02149524); EudraCT (2013-004172-35).

Natural magnetic nanoparticle containing droplet for smart drug delivery and heat treatment.

Biodegradable polymer droplet containing natural magnetic nanoparticle is composed for smart drug delivery and heat treatment. For selective and efficient drug delivery to the target tissue, direct high magnetic field will be applied near the target tissue. For drug release control and heat treatment, alternative high magnetic field will be applied. Magnetosome, natural magnetic nanoparticle, is extracted from magnetotactic bacteria, AMB-1. Mixture of magnetosome and sodium alginate composes into droplet using the microfluidic device applied Plateau-Rayleigh instability principle. The magnetosome contained droplet selected its rout at the bifurcate microchannels by direct high magnetic field. High alternative magnetic field generating circuit is designed with 18 mT and 4 Hz magnetic wave. The generated magnetic wave was applied to the extracted magnetosomes so that temperature of the magnetosomes increased from 15.2°C to 17.6°C.

In vitro and Lactation Responses in Mid-lactating Dairy Cows Fed Protected Amino Acids and Fat.

The objective of this study was to evaluate the effect of ruminally protected amino acids (RPAAs) and ruminally protected fat (RPF) supplementation on ruminal fermentation characteristics (in vitro) and milk yield and milk composition (in vivo). Fourteen mid-lactating Holstein dairy cows (mean weight 653±62.59 kg) were divided into two groups according to mean milk yield and number of days of postpartum. The cows were then fed a basal diet during adaptation (2 wk) and experimental diets during the treatment period (6 wk). Dietary treatments were i) a basal diet (control) and ii) basal diet containing 50 g of RPAAs (lysine and methionine, 3:1 ratio) and 50 g of RPF. In rumen fermentation trail (in vitro), RPAAs and RPF supplementation had no influence on the ruminal pH, dry matter digestibility, total volatile fatty acid production and ammonia-N concentration. In feeding trial (in vivo), milk yield (p<0.001), 4% fat corrected milk (p<0.05), milk fat (p<0.05), milk protein (p<0.001), and milk urea nitrogen (p<0.05) were greater in cows fed RPAAs and RPF than the corresponding values in the control group. With an index against as 0%, the rates of decrease in milk yield and milk protein were lower in RPAAs and RPF treated diet than those of basal diet group (p<0.05). In conclusion, diet supplemented with RPAAs and RPF can improve milk yield and milk composition without negatively affecting ruminal functions in Holstein dairy cows at mid-lactating.

Removal of As(V) from water by pectin based active hydrogels following geochemical approach.

Fe(II)-loaded pectin based hydrogels were used to remove As(V) from water. The hydrogels were synthesized by crosslinking pectin 'as such' or in the presence of 2-acrylamido-2-methylpropanesulphonic acid and three crosslinkers. The hydrogels crosslinked with 5% glutaraldehyde showed the maximum uptake of both Fe(II) and As(V). The sorption of As(V) was found to be sensitive to pH and temperature changes, as the maximum sorption was observed at 35 degrees C and 7.0-9.2 pH.

Investigation of refolding condition for Pseudomonas fluorescens lipase by response surface methodology.

Response Surface Methodology (RSM) was used to investigate optimal refolding conditions of Pseudomonas fluorescens lipase which was expressed as inclusion body in E. coli. Three interacting factors, protein concentration, pH and guanidine hydrochloride (GdnHCl) concentration were selected as the variables of RSM. The protein concentration did not affect the refolding yield within the selected range (50-340 micrograms ml-1) at low temperatures of 4 and -15 degrees C, but it was a critical factor at 25 degrees C and refolding yield significantly decreased with increasing protein concentration. The pH and GdnHCl were significant factors in all experimental conditions. But there was no trends of optimal pH depending on temperature and additives, just showing the optimum at neutral pH. Glycerol shifted optimal GdnHCl concentration to higher side, while arginine to lower side. Therefore, it was concluded that glycerol and arginine deprives and supplements GdnHCl, respectively. In this study, RSM made it possible to investigate successfully the optimal conditions of in vitro refolding and to elucidate interactions between refolding factors with a minimum number of experiments. Under the optimal condition determined by RSM, approximately 90% refolding yield was obtained and it was a 30% increase over the conventional method.

Inhibition of mutagenesis and transformation by root extracts of Panax ginseng in vitro.

The root extract of Panax ginseng was investigated for its inhibitory effects on DNA synthesis, mutagenicity, and cellular transformation using V79 and NIH 3T3 cells. DNA synthesis measured by the [3H]thymidine incorporation into V79 Chinese hamster lung cells was significantly decreased by the addition of ginseng extract (0-1 microgram/ml) to the medium. However, ginseng extract was found to increase the rate of DNA excision repair synthesis in V79 cells in response to treatment with UV radiation or methyl methanesulfonate. The extract also showed decreased mutation frequency when mutagenicity was examined using V79 cells at the hypoxanthine-guanine phosphoribosyl transferase locus as resistance to 6-thioguanine after exposure to methyl methanesulfonate. We also found that the components of ginseng extract continue to exert an inhibitory effect on the transformation of NIH 3T3 cells initiated by 3-methylchloanthrene, methyl methanesulfonate, and 1-methyl-3-nitro-1-nitrosoguanidine.