DNA-TCP complex structures reveal a unique recognition mechanism for TCP transcription factor families.

Plant-specific TCP transcription factors are key regulators of diverse plant functions. TCP transcription factors have long been annotated as basic helix-loop-helix (bHLH) transcription factors according to remote sequence homology without experimental validation, and their consensus DNA-binding sequences and protein-DNA recognition mechanisms have remained elusive. Here, we report the crystal structures of the class I TCP domain from AtTCP15 and the class II TCP domain from AtTCP10 in complex with different double-stranded DNA (dsDNA). The complex structures reveal that the TCP domain is a distinct DNA-binding motif and the homodimeric TCP domains adopt a unique three-site recognition mode, binding to dsDNA mainly through a central pair of ß-strands formed by the dimer interface and two basic flexible loops from each monomer. The consensus DNA-binding sequence for class I TCPs is a perfectly palindromic 11 bp (GTGGGNCCCAC), whereas that for class II TCPs is a near-palindromic 11 bp (GTGGTCCCCAC). The unique DNA binding mode allows the TCP domains to display broad specificity for a range of DNA sequences even shorter than 11 bp, adding further complexity to the regulatory network of plant TCP transcription factors.

Dimmed gene knockout shortens larval growth and reduces silk yield in the silkworm, Bombyx mori.

The bHLH domain transcription factor, Bombyx mori-derived dimmed (Bmdimm), is directly regulated by the JH-BmMet/BmSRC-BmKr-h1 pathway and plays a key role in regulating the expression of FibH, which codes the main component of silk protein. However, the other roles of Bmdimm in silk protein synthesis remain unclear. Here, we established a Bmdimm knockout (KO) line containing a 7-bp deletion via CRISPR/Cas9 system, which led to the absence of the bHLH domain. The expression level of silk protein genes and silk yield decreased significantly in the Bmdimm KO line. Moreover, knocking out Bmdimm led to shortened larval stages and significant weight loss in larvae and adults. Bmdimm was found to be highly expressed in the silk gland, but it was also expressed in the fat body. The expression level of Bmkr-h1 in the fat body was significantly downregulated in the Bmdimm KO line. Exogenous JHA treatment upregulated Bmkr-h1 and rescued the phenotype of larval growth in the Bmdimm KO line. In conclusion, knocking out Bmdimm led to a shortened larval stage via the inhibition of Bmkr-h1 expression, then reduced silk yield. These findings help to elucidate the regulatory mechanism of fibroin synthesis and larval development in silkworms.

Dilation of the cystic duct confluence in laparoscopic common bile duct exploration and stone extraction in patients with secondary choledocholithiasis.

BACKGROUND: Many options exist for the management of cholelithiasis and secondary choledocholithiasis. Among them, laparoscopic common bile duct exploration (LCBDE) with choledocotomy followed by laparoscopic cholecystectomy has gained popularity. However, efforts should be made to ensure minimally invasive or noninvasive management of the common bile duct (CBD). The purpose of this study was to explore the clinical experience of non-invasive surgical modality, i.e., laparoscopic transcystic dilation of the cystic duct confluence in CBD exploration (LTD-CBDE), including feasibility, safety, adverse events, and incidence. METHODS: In this retrospective analysis, 68 patients were offered the LTD-CBDE technique from December 2015 to April 2018 based on patient's own intention. During the surgery, the cystic duct confluence was dilated with separation forceps and/or a columnar dilation balloon. Subsequently, CBD exploration and stone extraction were performed with a choledochoscope. The entrance of the CBD was covered with a cystic duct stump wall and was subjected to primary closure at the end of surgery. RESULTS: Forty-nine females and 19 males with cholelithiasis and secondary choledocholithiasis were included. The mean age was 53 years old (18 to 72 year). Of these patients, 62 (91.2%) were successfully treated with the LTD-CBDE technique, and bile leakage was observed in 3 patients (4.4%). The mean operation time was 106 min, and the mean hospital stay was 5.9 days. Among the other 6 patients, 3 were converted to open cholecystectomy due to severe fibrosis, unclear anatomical structure at Calot's triangle (n = 2) or Mirizze syndrome (n = 1); LCBDE was performed in 3 patients due to cystic duct atresia (n = 2) and low level of flow from the gallbladder duct into the CBD (n = 1). These patients had a smooth postoperative course. In total, 43/68 of the patients presented no radiological evidence of retained CBD stones at the postoperative follow-up (40 patients treated with LTD-CBDE) 1 year later. CONCLUSIONS: The current work suggests that LTD-CBDE for the management of cholelithiasis and secondary choledocholithiasis is a feasible, safe and effective technique with a low complication rate. LTD-CBDE offers another alternative for surgeons to treat patients in similar scenarios. However, additional randomized, controlled studies are needed to demonstrate its efficacy, safety, and impact on CBD stenosis.

[Determination of recombinant hirudin in urine of rat by spectrophotometric method].

To develop a spectrophotometric method for determining the concentration of recombinant hirudin (rH) in urine of rats. rH concentration was determined based on the rH inhibility to thrombin which hydrolyzed the Chromozym TH TH chromogenic substrate to form the specific pNA absorbed at 405 nm. The standard rH in rat urine was determined by the spectrophotometric method at concentration of 6.25 to 75 ng x mL(-1) with day and intra-day RSD < 10%, method recoveries of > 95% and the dilution recoveries of > 93%. The rH samples of rat urines which iv dose of 0.5, 1.0, and 2.0 mg x kg(-1) were collected and analyzed by the CSA method. Their cumulative excretion rH at 0-12 hr were (116.850 +/- 57.160), (235.544 +/- 39.375) and (474.986 +/- 85.426) microg x kg(-1). The calculated cumulative excretion rate of three doses is about 23% which indicates that the rH was eliminated in the way of a linear kinetics in rats. The rH content in rat urine could be measured by the spectrophotometric method accurately, reliably and sensitively for the rH urinary excretion dynamics study.

Transcriptome analysis reveals key transcription factors and pathways of polian vesicle associated with cell proliferation in Vibrio splendidus-challenged Apostichopus japonicus.

Polian vesicle is thought to produce coelomocytes and contribute to the sea cucumber's immune system. Our previous work has indicated that polian vesicle was responsible for cell proliferation at 72 h post pathogenic challenge. However, the transcription factors related to the activation of effector factors and the molecular process behind this remained unknown. In this study, to reveal the early functions of polian vesicle in response to the microbe, a comparative transcriptome sequencing of polian vesicle in V. splendidus-challenged Apostichopus japonicus, including normal group (PV 0 h), pathogen challenging for 6 h (PV 6 h) and 12 h (PV 12 h) was performed. Compared PV 0 h to PV 6 h, PV 0 h to PV 12 h, and PV 6 h to PV 12 h, we found 69, 211, and 175 differentially expressed genes (DEGs), respectively. KEGG enrichment analysis revealed the DEGs, including several transcription factors such as fos, FOS-FOX, ATF2, egr1, KLF2, and Notch3 between PV 6 h and PV 12 h were consistently enriched in MAPK, Apelin and Notch3 signaling pathways related to cell proliferation compared with that in PV 0 h. Important DEGs involved in cell growth were chosen, and their expression patterns were almost the same as the transcriptome profile analysis by qPCR. Protein interaction network analysis indicated that two DEGs of fos and egr1 were probably significant as key candidate genes controlling cell proliferation and differentiation in polian vesicle after pathogenic infection in A. japonicus. Overall, our analysis demonstrates that polian vesicles may play an essential role in regulating proliferation via transcription factors-mediated signaling pathway in A. japonicus and provide new insights into hematopoietic modulation of polian vesicles in response to pathogen infection.

Antitumor activity of lobaplatin alone or in combination with antitubulin agents in non-small-cell lung cancer.

OBJECTIVE: Lobaplatin is used to treat patients with breast cancer, small-cell lung cancer, and chronic myelogenous leukemia in China. In this study, we assessed the in-vitro and in-vivo activities of lobaplatin alone or in combination with antitubulin agents against human non-small-cell lung cancer (NSCLC). METHODS: The cytotoxicities of lobaplatin against NSCLC cells were determined by the sulforhodamine B (SRB) assay. Cell cycle analysis and apoptosis were assessed using flow cytometry, and the in-vivo antitumor activities were evaluated in human NSCLC xenografts in nude mice. RESULTS: The cytotoxicity of lobaplatin was similar to or higher than that of cisplatin and carboplatin, with a mean IC(50) of 2.5 µmol/l in a variety of NSCLC cells. In addition, lobaplatin arrested cells in the S phase and triggered apoptosis. The combination of lobaplatin with antitubulin agents yielded synergistic cytotoxic activity in vitro. In NSCLC xenografts, lobaplatin alone showed significant antitumor activity. The combination of lobaplatin with antitubulin agents, especially with paclitaxel, led to significantly enhanced activity, which was superior to that of cisplatin combined with antitubulin agents. CONCLUSION: These data demonstrate that the use of lobaplatin alone or in combination with antitubulin agents might be a rational and novel therapeutic strategy for human NSCLC and warrants further clinical investigation.

Neuroprotective properties of aucubin in diabetic rats and diabetic encephalopathy rats.

In this study, we determined the neuroprotective effect of aucubin on diabetes and diabetic encephalopathy. With the exception of the control group, all rats received intraperitoneal injections of streptozotocin (STZ; 60 mg/kg) to induce type 1 diabetes mellitus (DM). Aucubin (1, 5, 10 mg/kg ip) was used after induction of DM (immediately) and diabetic encephalopathy (65 days after the induction of diabetes). The diabetic encephalopathy treatment groups were divided into short-term and long-term treatment groups. Treatment responses to all parameters were examined (body weight, plasma glucose, Y-maze error rates and proportion of apoptotic cells). In diabetic rats, aucubin controlled blood glucose levels effectively, prevented complications, and improved the quality of life of diabetic rats. In diabetic encephalopathy, aucubin significantly rescued neurons in the hippocampal CA1 subfield and reduced working errors during behavioral testing. The significant neuroprotective effect of aucubin could be seen not only in the short term (15 days) but also in the long term (45 days), which was a highly encouraging finding. These data suggest that aucubin may be a potential neuroprotective agent.

Protective effects of aucubin on H2O2-induced apoptosis in PC12 cells.

The present study investigated the neuroprotective effects of aucubin on hydrogen peroxide (H2O2)-induced apoptosis in PC12 cells. Exposure of PC12 cells to 0.25 mm H2O2 induced a leakage of lactate dehydrogenase and decreased cell viability, as shown by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. In a dose over 0.1 mm, aucubin increased PC12 cellular viability and markedly attenuated H2O2-induced apoptotic cell death. Quantitation of apoptosis by flow cytometry indicated that aucubin inhibited H2O2-induced apoptosis in PC12 cells. Nuclear damage was alleviated by aucubin, as shown by Hoechst staining. In addition, the levels of malondialdehyde were reduced and the activity of superoxide dismutase, catalase and glutathione peroxidase was augmented in these cells. These results indicated that aucubin inhibited H2O2-induced apoptosis in PC12 cells through regulation of the endogenous oxidant-antioxidant balance. Our results suggest that aucubin is a potential protective agent for the treatment of oxidative-stress-induced neurodegenerative disease.

Occurrence of antibiotics in waters, removal by microalgae-based systems, and their toxicological effects: A review.

Global antibiotics consumption has been on the rise, leading to increased antibiotics release into the environment, which threatens public health by selecting for antibiotic resistant bacteria and resistance genes, and may endanger the entire ecosystem by impairing primary production. Conventional bacteria-based treatment methods are only moderately effective in antibiotics removal, while abiotic approaches such as advanced oxidation and adsorption are costly and energy/chemical intensive, and may cause secondary pollution. Considered as a promising alternative, microalgae-based technology requires no extra chemical addition, and can realize tremendous CO2 mitigation accompanying growth related pollutants removal. Previous studies on microalgae-based antibiotics removal, however, focused more on the removal performances than on the removal mechanisms, and few studies have concerned the toxicity of antibiotics to microalgae during the treatment process. Yet understanding the removal mechanisms can be of great help for targeted microalgae-based antibiotics removal performances improvement. Moreover, most of the removal and toxicity studies were carried out using environment-irrelevant high concentrations of antibiotics, leading to reduced guidance for real-world situations. Integrating the two research fields can be helpful for both improving antibiotics removal and avoiding toxicological effects to primary producers by the residual pollutants. This study, therefore, aims to build a link connecting the occurrence of antibiotics in the aquatic environment, the removal of antibiotics by microalgae-based processes, and the toxicity of antibiotics to microalgae. Distribution of various categories of antibiotics in different water environments were summarized, together with the antibiotics removal mechanisms and performances in microalgae-based systems, and the toxicological mechanisms and toxicity of antibiotics to microalgae after either short-term or long-term exposure. Current research gaps and future prospects were also analyzed. The review could provide much valuable information to the related fields, and provoke interesting thoughts on integrating microalgae-based antibiotics removal research and toxicity research on the basis of environmentally relevant concentrations.

Differentiation and characteristics of the enhanced green fluorescent protein gene transgenic goat neural stem cells cultured in attached and non-attached plates.

The aims of this study were (i) to determine whether NSCs (neural stem cells) could be isolated from the brain of embryonic day 98 fetal goat, (ii) to determine if these stem cells have the capability of multipotent differentiation following transfection with a reporter gene, EGFP (enhanced green fluorescent protein) and (iii) to study the characteristics of the stem cells cultured in attached and non-attached plates. NSCs were isolated from embryonic day 98 fetal goat brain, transfected with EGFP gene using lipofection, and subcultured in attached and non-attached plates respectively. The transgenic stem cells were induced to differentiate into osteogenic and endothelial cells in vitro respectively. Markers associated with undifferentiated NSCs and their differentiated cells were tested by RT-PCR (reverse transcription-PCR). The results demonstrated that stem cells could be isolated from embryonic day 98 fetal goat brain, and EGFP gene could be transfected into the cells. The transgenic NSCs were capable of self-renewal, a defining property of stem cells, and were grown as free-floating neurospheres in non-attached plates. When the neurospheres were transferred and cultured in attached plates, cells migrate from the neurospheres and are grown as spindle cells. The stem cells were grown as quasi-circular cells when the single stem cells were cultured in attached plates. Both the NSCs cultured in non-attached and attached plates could express Hes1 (hairy and enhancer of split 1), Oct4 (octamer-binding protein 4), Nanog, Sox2 [SRY (sex-determining region Y)-box 2] and Nestin, while following differentiation cells expressed markers for osteogenic cells (Osteocalcin+ and Osteonectin+) and endothelium (CD34+ and eNOS+). The results demonstrated that the goat EGFP gene transgenic NSCs have the capability of multipotent differentiation, which means that the transgenic NSCs may be useful in cell transplantation studies in future.

Differentiation of AFS cells derived from the EGFP gene transgenic porcine fetuses.

We have obtained the EGFP (enhanced green fluorescence protein) gene transgenic porcine fetuses before. The aims of this study were (i) to determine whether stem cells could be isolated from amniotic fluid of the transgenic porcine fetuses, and (ii) to determine if these stem cells could express EGFP and differentiate in vitro. The results demonstrated that stem cells could be isolated from amniotic fluid of the EGFP gene transgenic porcine fetuses and could express EGFP and differentiate in vitro. Undifferentiated AFSs (amniotic fluid-derived stem cells) expressed POU5F1, THY1 and SOX2, while the following differentiation cells expressed markers for chondrogenic (COL2A1), osteogenic (osteocalcin and osteonectin) and neurogenic cells such as astrocyte (GFAP), oligodendrocyte (GALC) and neuron (NF, ENO2 and MAP).

Aucubin modulates Bcl-2 family proteins expression and inhibits caspases cascade in H2O2-induced PC12 cells.

In this study, the effect of aucubin on H(2)O(2)-induced apoptosis was studied by using a rat pheochromocytoma (PC12) cell line. We have analyzed the apoptosis of H(2)O(2)-induced PC12 cells, H(2)O(2)-induced apoptosis appeared to correlate with lower Bcl-2 expression, higher Bax expression and sequential activation of caspase-3 leading to cleavage of poly-ADP-ribose polymerase (PARP). Aucubin not only inhibited lower Bcl-2 expression, high Bax expression, but also modulated caspase-3 activation, PARP cleavage, and eventually protected against H(2)O(2)-induced apoptosis. These results indicated that aucubin can obstruct H(2)O(2)-induced apoptosis by regulating of the expression of Bcl-2 and Bax, as well as suppression of caspases cascade activation.

Expression of vascular endothelial growth factor C correlates with a poor prognosis based on analysis of prognostic factors in patients with cervical carcinomas.

AIM: The aim of this study was to specifically investigate the clinicopathological role of expression of vascular endothelial growth factor C (VEGF-C) as well as the correlation with clinical outcomes in cervical cancer. METHOD: We carried out a comparative analysis of data from a prospective observational study of 82 patients with cervical carcinomas who underwent radical hysterectomy with bilateral adnexectomy and bilateral pelvic lymphadenectomy from January 2001 to January 2005. RESULTS: VEGF-C mRNA was identified in tumor tissues from 42 patients. In univariate analysis by Fisher's exact probability test, VEGF-C expression was significantly associated with lymph node metastasis (LNM) (pN1) (P < 0.01). In multivariate analysis by Binary logistic regression analysis, LNM was the independent relevant factor for VEGF-C mRNA expression in tumor tissues. In univariate analysis by the log-rank test, the overall 5-year survival rate of the patients with VEGF-C mRNA expression in tumor tissues was significantly lower than that of the patients without VEGF-C mRNA expression (47.6% vs. 87.5%; P < 0.01). The overall 5-year survival rate of the patients with LNM was significantly lower than that of the patients without LNM (50.0% vs 80.4%; P < 0.01). According to Cox regression multivariate analysis, VEGF-C mRNA expression in tumor tissues and LNM were independent, relevant factors for 5-year survival rate, respectively. CONCLUSIONS: Expression of VEGF-C is related to lymph node metastasis (pN1), and is a prognostic indicator for cervical cancer.

Comparison on nutritional quality of adult female Chinese mitten crab (Eriocheir sinensis) with different colored hepatopancreases.

White hepatopancreas syndrome has recently emerged in Chinese mitten crab (Eriocheir sinensis) aquaculture, causing considerable economic loss. The hepatopancreas color of diseased crabs becomes gradually lighter, turning from yellow to yellow-white to white. Therefore, this study was conducted to investigate the changes in nutrient composition in three edible parts (hepatopancreas, ovaries, and muscle) of adult females with different colored hepatopancreases. Three groups were assessed in this study, including a yellow hepatopancreas group (control, L * = 63.92, a * = 22.14, b * = 60.95), a yellow-white hepatopancreas group (YWHG, L * = 65.06, a * = 22.35, b * = 57.80), and a white hepatopancreas group (WHG, L * = 65.72, a * = 10.70, b * = 30.52). No statistically significant differences in average weight, tissue indices, and total edible yield were observed among the three crab groups (P >0.05). The moisture content of the hepatopancreases and ovaries in WHG was 56.12% and 9.23% higher than the control values (P <0.05), whereas hepatopancreas crude fat and ovary crude protein levels were 62.23% and 11.45% lower than the control values (P < 0.05). The total carbohydrate levels of the three edible tissues were significantly higher and the crude protein content of ovaries was significantly lower in YWHG (P < 0.05). Most amino acid levels in the WHG muscle and ovaries were significantly lower than the control (P < 0.05). Moreover, the hepatopancreas levels of total polyunsaturated fatty acids (PUFA) and n-6PUFA in WHG were 24.88% and 31.83% lower than in control group, whereas the hepatopancreas levels of total PUFA and n-6PUFA in YWHG were also 21.88% and 23.20% lower compared to the controls (P < 0.05). Overall, the growth and the edible parts were not affected in YWHG and WHG. Moreover, YWHG crabs exhibited few effects on nutritional value; however, the fatty acid composition of crabs was significantly changed. In contrast, WHG crabs exhibited poor nutritional quality. Nonetheless, the consumption of crabs with yellow-white or white hepatopancreases is not recommended since the animal also referred to as diseased crabs.

Efficacy of specific egg yolk immunoglobulin (IgY) to bovine mastitis caused by Staphylococcus aureus.

The objective of this study was to estimate the efficacy of specific egg yolk immunoglobulin (IgY) to bovine mastitis caused by Staphylococcus aureus. Eighteen lactating cows with clinical mastitis and 18 lactating cows with experimental mastitis (1 quarter per cow) were randomly assigned to three treatments: IgY (20mg/ml) infusion, penicillin (100mg/ml) infusion and no infusion. Treatments for clinical mastitis and experimental mastitis were performed by a 6-day course of intramammary infusion with a dosage of 10ml at an interval of 12h. Milk samples were collected at morning milking time for testing color, clot, somatic cell counts (SCC) and bacterial count. For most of the cows treated with IgY and penicillin, the milk color and clot recovered to normal form during the therapy course. The milk SCCs and bacterial counts of treated cows decreased compared to those of untreated cows (p<0.05). The cure rates by IgY for experimental and clinical mastitis were 83.3% and 50%, respectively, and those by penicillin were 66.7% and 33.3%, respectively. These results showed the potential of specific IgY to be an alternative therapy for mastitis caused by S. aureus.

Chitosan-alginate microcapsules for oral delivery of egg yolk immunoglobulin (IgY): in vivo evaluation in a pig model of enteric colibacillosis.

In our previous study, the applicability of chitosan-alginate microcapsules for oral delivery of egg yolk immunoglobulin (IgY) was established in a simulated gastrointestinal tract environment. The objective of the present study was to evaluate the protective efficacy of microencapsulated IgY against K88+ ETEC (enterotoxigenic Escherichia coli)-induced diarrhea in 40-day-old pigs. Groups of pigs orally challenged with 10(11) cfu/mL of K88+ ETEC were fed with non-encapsulated IgY, microencapsulated IgY and aureomycin-treated feed respectively. The clinical response of each group was monitored and evaluated in terms of lethargy, inappetence, occurrence of diarrhea, fecal consistency score, weight loss and recovery rate. The results showed that treatment of infected pigs with microencapsulated IgY significantly (P<0.05) reduced the K88+ ETEC-induced diarrhea at 24 h post-infection. In contrast, the diarrhea-reducing effect of non-encapsulated IgY was delayed (only evident after 72 h) while normal saline-treated pigs (controls) continued to suffer from diarrhea and dehydration. Similarly, weight gain in microencapsulated IgY-treated pigs was better and significantly different (P<0.05) than in non-encapsulated IgY and saline-treated controls. Collectively, these results support previous in vitro observations showing that chitosan-alginate microcapsules can be an effective method of protecting IgY from gastric inactivation, enabling its use for the widespread prevention and control of enteric diseases.

Aucubin prevents loss of hippocampal neurons and regulates antioxidative activity in diabetic encephalopathy rats.

In this study, the neuroprotection of aucubin and its mechanism were evaluated in the rat model of diabetic encephalopathy. Diabetes mellitus (DM) rats were stratified by cognitive capability (CC), and assigned to four treatment groups for aucubin treatment (doses of 0, 1, 5 or 10 mg/kg aucubin), with a further two groups of non-DM rats ranked by CC as controls for aucubin (doses of 0 or 5 mg/kg aucubin). Neuroprotection was estimated by the indexes of behavior and histology. Behavioral testing was performed in a Y-maze. The surviving neurons in CA1-CA4 and subiculum (SC) of the hippocampus were counted under a microscope. In addition, the apoptotic neurons in the CA1 of the hippocampus were also examined by using TUNEL staining. In order to clarify the mechanism of aucubin's neuroprotection, the activities of endogenous antioxidants and nitric oxide synthase (NOS) together with the content of lipid peroxide in the hippocampus were assayed. The results proved that aucubin significantly reduced the content of lipid peroxide, regulated the activities of antioxidant enzymatic and decreased the activity of NOS. All these effects indicated that aucubin was a potential neuroprotective agent and its neuroprotective effects were achieved, at least in part, by promoting endogenous antioxidant enzymatic activities.

Diversity and Dynamics of Yeasts During Vidal Blanc Icewine Fermentation: A Strategy of the Combination of Culture-Dependent and High-Throughput Sequencing Approaches.

In this study, attention has been focused on the ecology of yeasts during the spontaneous and inoculated fermentation processes of Vidal blanc icewine in northeast China, which is very important for screening autochthonous yeast strains, understanding the roles of these strains, and managing fermentation. The strategies were to conduct spontaneous and inoculated laboratory-scale fermentation processes simultaneously and to analyze the samples taken at different fermentation stages by culture-dependent and -independent methods. Three hundred and thirty-eight yeast strains were isolated and twelve genera were identified by sequencing. During the spontaneous fermentation process, non-Saccharomyces yeasts were predominant in the initial and middle stages, whereas Saccharomyces dominated in the later stages; Candida was preponderant in the whole process, and its abundance in the final stages was only lower than Saccharomyces. The inoculated fermentation was characterized by a predominance of Saccharomyces throughout the fermentation process; non-Saccharomyces yeasts were observed in the early stage. The internal transcribed spacer (ITS) 2 region gene was firstly used to analyze the yeast diversity in the samples during the icewine fermentation processes by high-throughput sequencing (HTS), and a more complex mycobiota was revealed. Moreover, the dynamics of other major fungi (mainly Davidiella and Alternaria) during icewine fermentation processes were also revealed, which have never been reported in icewine before.

Antioxidant and pancreas-protective effect of aucubin on rats with streptozotocin-induced diabetes.

Oxidative stress has been suggested as a contributory factor in development and complication of diabetes. The aim of the present study was to determine the protective effect of aucubin on lipid peroxidation and activities of antioxidant defense systems and to conduct immunohistochemical evaluation of pancreas in streptozotocin-induced diabetic rats. Lipid peroxidation was determined by assessing the concentration of malondialdehyde and activities of antioxidant enzymes - catalase, glutathione peroxidase and superoxide dismutase in liver and kidneys of rats were determined. Changes of blood glucose and immunohistochemical evaluation on pancreas were also investigated as part of the pathology of diabetes. In our study, aucubin treatment lowered blood glucose. Diabetic rats exhibited an increase in the level of lipid peroxidation and decrease in activities of antioxidant enzymes in liver and kidneys as compared to control rats. Administration of aucubin to diabetic rats for 15 days significantly reversed damage associated with diabetes. In addition, diabetic rats showed an obvious decrease in insulin immunoreactivity and the number of beta cells in pancreas, but the pancreas of aucubin-treated rats were improved and the number of immunoreactive beta cells were significantly increased. These results indicated that aucubin may have value as a safe preventive or therapeutic agent against diabetes mellitus.

Characterization of specific egg yolk immunoglobulin (IgY) against mastitis-causing Escherichia coli.

The objective of this study was to estimate the in vitro activity of egg yolk immunoglobulin (IgY) against mastitis-causing Escherichia coli. Specific IgY was produced by hens immunized with formaldehyde killed E. coli O111 in long-standing immunization response (titer > or =6400 for 100 days) and was isolated from yolks with a purity of 86% by water dilution, salt precipitations and ultrafiltration. Enzyme-linked immunosorbent assay (ELISA) indicated the produced IgY specifically targeted E. coli O111 and five other E. coli strains which were isolated from mastitic cows. The growth inhibition activity of the specific IgY to bacteria was dose-dependent with an effective concentration of 20mg purified IgY per milliliter. The phagocytic activity of E. coli either by milk macrophages (MPhi) or by polymorphonuclear neutrophil leukocytes (PMN) in the presence of specific IgY was significantly higher than that with nonspecific IgY or without IgY (p<0.05), suggesting that it enhanced phagocytic activity. The current work suggests that this specific IgY has potential as a therapeutic treatment for mastitis in dairy cows.