Tartary buckwheat has established itself as a functional food source because of its basic nutrition and phenolic compound contents, such as dietary fiber (DF) and rutin (RU). However, little information has been obtained concerning the comparative effects of DF and RU on the in vitro and in vivo glucose responses of tartary buckwheat flour. Moreover, the relationship between the flour’s in vitro starch digestibility and its components’ blood glucose response is not well-known. This study found that DF and RU reduced rapidly digestible starch (RDS) by 37.32→33.88% and 41.71→30.28%, whereas they increased resistant starch (RS) by 30.47→31.46% and 28.41→36.78%, respectively. Furthermore, RU had a lower glycemic index (GI) compared to DF. The regression equation for the in vitro and in vivo data from RU exhibited positive correlation (R 2 = 0.99); however, DF did not display positive correlation, which indicates that the in vitro and in vivo GI mechanisms by DF and RU are different.

The hand held voltammetry systems searched diabetic assay using glucose sensor of fluorine nafion doped carbon nanotube electrode (FCNE). An inexpensive graphite carbon pencil was used as an Ag/AgCl reference and Pt counter electrode. Upon combining and using three electrode systems, optimum square wave (SW) stripping results were attained to 1.0-9.0 ug/L with 8 points. Statistic RSD precision was of 6.02 % with n=15 in 0.1 mg/L glucose. After a total of 200 second accumulation times, analytical detection limit of 0.8 ug/L was obtained. This developed technique was applied to urine samples from diabetic patients urine for fluid analysis, it was determined that the sensor can be used with a diagnostics in the ex vivo of live cells and non treated biological fluid.

Zona pellucida (ZP), a primarily representative coat of mammalian egg and embryo, has an extremely heterogeneous morphology during different developmental stages. The objective of the present study was to compare the morphological changes of the ZP surface of immature, in vitro and in vivo matured canine oocytes by using scanning electron microscopy (SEM). Canine ovaries were collected from local veterinary hospitals to recover immature oocytes. The ovaries were sliced and the released cumulus oocyte complexes (COCs) were washed with TL-HEPES. The selected COCs were randomly divided into two groups, first group was processed immediately at immature state and the second group was processed 72 h after in vitro maturation, and compared with in vivo derived oocytes. Oocytes were fixed, critical point dried and examined under SEM. The diameters of oocyte and outer holes of the ZP were measured on a total of 249 oocytes; the results were analyzed using One-way ANOVA. Our results showed that, the diameter of immature oocytes significantly differed (p < 0.05) from that of in vivo matured oocytes (79.60 ± 0.77 μm vs. 101.46 ± 1.07 μm, respectively). Similarly, a significant difference (p < 0.05) in the diameters between those of in vitro and in vivo matured oocytes were found (79.51 ± 2.36 μm vs. 101.46 ± 1.07 μm, respectively). Moreover, the diameters of the outer holes of the ZP were significantly (p < 0.05) larger in in vivo matured (1.48 ± 0.42 μm) than in vitro matured for 72 and immature oocytes (1.10 ± 0.16 and 0.43 ± 0.12 μm, respectively). Taken together, these data indicates that the ZP surface is related to oocyte maturity in canine.

Polo-like kinase 1 (Plk1) has been known to be a critical element in cell division including centrosome maturation, cytokinesis and spindle formation in somatic, cancer, and mammalian embryonic cells. In particular, Plk1 is highly expressed in cancer cells. Plk1 inhibitors, such as BI2536, have been widely used to prevent cell division as an anticancer drug. In this study, the fertilized murine oocytes were treated with BI2536 for 30 min after recovery from the oviduct to investigate the effect of down-regulation of Plk1 in the in vivo-fertilized murine embryos. Then, the localization and expression of Plk1 was observed by immunofluorescence staining. The sperm which had entered into the oocyte cytoplasm did not form male pronuclei in BI2536-treated oocytes. The BI2536-treated oocytes showed significantly lower expression of Plk1 than non-treated control group. In addition, alpha-tubulin and Plk1 gathered around sperm head in non-treated oocytes, while BI2536-treated oocytes did not show this phenomenon. The present study demonstrates that the Plk1 inhibitor, BI2536, hinders fertilization by inhibiting the formation of murine male pronucleus

Porphyromonas gingivalis is among the major etiological pathogens of chronic periodontitis. The virulence mechanisms of P. gingivalis is yet to be identified as its activity is largely unknown in actual disease process. The purpose of this study is to identify antigens of P. gingivalis expressed only in patients with chronic periodontitis using a unique immunoscreening technique. Change Mediated Antigen Technology (CMAT), an antibody-based screening technique, was used to identify virulence-associated proteins of P. gingivalis that are expressed only during infection stage in patients having chronic periodontitis. Out of 13,000 recombinant clones screened, 22 tested positive for reproducible reactivity with rabbit hyperimmune anti-sera prepared against dental plaque samples acquired from periodontitis patients. The DNA sequences of these 18 genes were determined. CMAT-identified protein antigens of P. gingivalis included proteins involved in energy metabolism and biosynthesis, heme and iron binding, drug resistance, specific enzyme activities, and unknown functions. Further analysis of these genes could result in a novel insight into the virulence mechanisms of P. gingivalis.

The present study was to assess the in vivo embryo production efficiency using the semen separated according to sex during superovulation in Hanwoo. Seventy Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered on 7 days after the third insemination by flushing the uterus with embryo collection medium. KPN semen straws used artificial insemination contained 20 million sperm (total number 60 million per donor). Sex-sorted semen straws contained 4 million sperm (total number 12 million per donor). The results obtained were as follows: No differences were observed in the efficiency of superovulation rates on KPN semen 87%, and sexed semen 100%, respectively. The mean numbers of total embryos are each 12.58 ± 8.31 and 13.25 ± 7.86. The mean numbers of transferable embryos, sexed semen were significantly lower than KPN semen (3.75 ± 1.98 vs. 8.23 ± 6.07, P<0.05). The rates of unfertilized embryos from superovulation using sexed semen were significantly higher than KPN semen (50% vs. 15%, P<0.05). The rate of degenerated 2-cell embryos from sexed and KPN semen was 60.87% and 11.11%, respectively (p<0.05). In conclusion, these results indicate that superovulation using sexed semen was useful, but efficient embryo production was important to reducing the damage caused by the Flowcytometer-based sperm sorting procedure.

Transplantation of stem cells, such as mesenchymal stem cells (MSCs), is a promising strategy for treating several types of intractable disorders. Mechanistically, it could not only replace damaged cells by direct contribution, but also establish an anti-inflammatory or immunomodulatory microenvironment. However, the cellular mechanisms underlying molecular and biological properties of stem cells during ex vivo expansion and also after transplantation in pathological environments remain largely elusive. We recently developed the cyanoacrylamide-based coumarin derivatives (Fluorescent real-time thiol tracer; FreSHtracer*) reversibly react with glutathione for monitoring of glutathione levels in living stem cells. These probes revealed that glutathione levels are heterogeneous among subcellular organelles and among individual cells and show dynamic changes and heterogeneity in repopulating stem cells depending on oxidative-stress or culture conditions. Importantly, a subpopulation of stem cells with high-glutathione levels exhibited increased self-renewal and migration activities in vitro and showed improved therapeutic efficiency in treating asthma. Furthermore, employing a novel combination of longitudinal intravital confocal fluorescence imaging and microcystoscopy in living animals, we investigated the distributions and properties of transplanted multipotent MSCs derived from human embryonic stem cells at single-cell resolution in real-time by performing confocal imaging of bladder tissues in a rat model of IC/BPS for up to 6 months post-transplantation. These novel real-time monitoring strategies demonstrate the novel molecular insight for maintaining stem cell functions and also enhance understanding of the in vivo behaviors of the engrafted stem cells, which is crucial to determine the efficacy and safety of stem cell-based therapies. This strategy may facilitate the translation of various stem cell-based approaches into clinical practice.

Nosema disease caused by the microsporidia Nosema apis and Nosema ceranae are a honey bee pathogen parasitizing. Nosema disease symptoms include digestive and absorption disorders because the spores damage epithelial tissue and potentially causing colony death. Recently, N. ceranae has been reported as an important threat to honey bee health. Turmeric (Curcuma longa L.) Curcuma tonga L. belongs to the family Zingiberaceae and is a perennial, tropical herb. Turmeric, the powdered rhizome, is used for medicinal purposes. The aim of this study was to evaluate the potential of Turmeric (Curcuma longa L.) for the control of N. ceranae in honeybees. For the study, we infected with N. ceranae spore through dosed and fed with the turmeric extraction at difference concentration. The data show that the turmeric extraction was not toxic for bee at least at 1% and the bees fed with 0.5 % turmeric extraction had significantly lower infection rates. This data suggest that turmeric could be useful in alternative strategies for the control of N. ceranae.

Salvia miltiorrhiza (Danshen) is perennial plant and commonly used in traditional Chinese medicine to treat numerous diseases like cardiovascular and cerebrovascular diseases, coronary heart disease, myocardial infarction, stroke and some viral diseases. Nevertheless, no study has been conducted on the antiviral properties of crude extract of Salvia miltiorrhiza against Influenza virus. In an attempt to identify new potential anti-influenza virus agents, 200 natural oriental herbal medicines were screened and we found that Salvia miltiorrhiza has a potential anti-influenza effect. Therefore, in this study, we investigated the protective effect of aqueous extract from Salvia miltiorrhiza against divergent influenza A subtypes using murine model of influenza A infection. Effective dose of aqueous extracts of Salvia miltiorrhiza in BALB/c mice displayed higher survival rate and lower lung viral titers when challenged with lethal doses of influenza A subtypes ({A/Aquatic bird/Korea/ W81/2005(H5N2)}, {A/PR/8/34(H1N1)}, {A/Aquatic bird/Korea/W44/2005(H7N3)} and {A/Chicken/ Korea/116/2004(H9N2)}). In vivo results exhibited that Salvia miltiorrhiza induced prophylactic effect in BALB/c mice against Influenza virus by disrupting viral replication or preventing viral infection by creating an antiviral state in the lungs. Taken together, the use of aqueous extracts of Salvia miltiorrhiza as an orally active antiviral agent, will be potential candidates for prophylactic treatments against Influenza A virus for humans and animals.

In the current study, we investigated the inhibitory activity of water soluble β-glucan from oat (Avena sativa) against various digestive enzymes such as α-glucosidase, sucrase, maltase and glucoamylase. Inhibition of these enzymes involved in the absorption of disaccharide can significantly decrease the post-prandial increase of blood glucose level after a mixed carbohydrate diet. The β-glucan had the highest documented rate of small intestinal sucrase inhibitory activity (2.83 mg/mL, IC50) relevant for potentially managing post-prandial hyperglycemia. Furthermore, we evaluated the effects of β-glucan on the level of post-prandial blood glucose in animal model. The post-prandial blood glucose levels were tested two hours after sucrose/starch administration, with and without β- glucan (100, and 500 mg/kg-body weight). The maximum blood glucose levels (Cmax) of β-glucan administration group were decreased by about 23% (from 219.06±27.82 to 190.44±13.18, p<0.05) and 10% (from 182.44±13.77 to 165.64±10.59, p<0.01) in starch and sucrose loading test, respectively, when compared to control in pharmacodynamics study. The β -Glucan administration significantly lowered the mean, maximum, and minimum level of post-prandial blood glucose at 30 min after meal. In view of the foregoing, it is felt that our findings suggest that β-glucan from oat serves to reduce post-prandial blood glucose rise secondary to slower absorption of glucose in the small intestine, via carbohydrate hydrolyzing enzymes inhibition.