Preparation and characterization of Antarctic krill oil/quercetin co-loaded liposomes and their protective effect on oleic acid-induced steatosis and oxidative stress in vitro.

This study aims to introduce a new liposome to co-load Antarctic krill oil (AKO) and quercetin (QC) as a new delivery formulation to enrich the application of AKO and QC. The stability of liposomes could be increased by adding an appropriate quantity of soy lecithin (SL). Changes in the composition of the phospholipid membrane were strongly correlated with the stability and release capacity of loaded nutrients. SL2@QC/AKO-lips displayed a nearly spherical shape with higher oxidative stability and controlled the in vitro release performance of QC in simulated digestion. Moreover, in vitro studies indicated that new liposomes had no adverse effects on cell viability and could combine the physiological functions of AKO and QC to protect the HepG2 cells from oleic acid-induced steatosis and oxidative stress. The findings demonstrated that the AKO and QC co-loaded liposomes prepared with the addition of an appropriate quantity of SL had excellent loading efficiency of AKO/QC and good oxidative stability, security and functional activity.

The effects of oil type and crystallization temperature on the physical properties of vitamin C-loaded oleogels prepared by an emulsion-templated approach.

Vitamin C (VC) is widely used as an antioxidant and nutrient to increase the nutritional value and shelf-life of foods. In this article, VC was loaded in oleogels using a simple two-step emulsion-templated approach and the effects of oil type (linseed oil, corn oil, and camellia oil) and crystallization temperature (Tc, -18, 0, 5, and 25 °C) on the physical properties, VC concentration, and oxidation stability of the VC-loaded oleogels were studied. As the amount of saturated fatty acids in the oil phase of the oleogels decreased, the VC loading level, oxidation stability and physical properties of the corn-oil-based oleogel (COG) were better than those of camellia-oil-based oleogels and linseed-oil-based oleogels. At different Tc values, the temperature and frequency dependent storage modulus values for the COG crystallized at 0 °C and 5 °C were not significantly different (P > 0.05), but their values were higher than those for COG crystallized at -18 °C and 25 °C (P < 0.05); the firmness of the COG crystallized at -18 °C and 0 °C was higher than those crystallized at 5 °C and 25 °C (P < 0.05); the network of the COG crystallized at 0 °C was denser than those of the COG crystallized at -18 °C, 5 °C, and 25 °C; and the VC concentration of the oleogels was affected by the crystallization temperature (Tc) and temperature fluctuations. To sum up, a VC-loaded oleogel with excellent mechanical properties was prepared using corn oil and crystallized at 0 °C via an emulsion-templated approach.

Effect of Palm Stearin Oil (PSO), Physical Fractionation Oil of Palm Stearin Oil (PPP) and Structured Lipids of Rich 1,3-Dioleoyl-2-palmitoylglycerol (OPO)on the Stability in Different Emulsion Systems.

This manuscript described the preparation of triglycerides with palmitic and ethyl oleate chains, and the stability of emulsions prepared from those triglycerides. Results showed that ratios of total saturated fatty acids (ΣSFA) of palm stearin oil (PSO), physical fractionation oil of palm stearin oil (PPP) and structured lipids of rich 1,3-dioleoyl-2-palmitoylglycerol (OPO) were 72.5%, 95.4% and 33.2% respectively. Rich 1,3-dioleoyl-2 palmitoylglycerol-emulsion (OPO-E) showed a better emulsion stability than that of palm stearin oil (PSO) and physical fractionation oil of palm stearin oil (PPP). The emulsion stability of OPO-E with 10% structured lipids of rich 1,3-dioleoyl-2-palmitoylglycerol (OPO) showed the highest value compared with 5% and 20% OPO. The value of emulsion stability (ES) was 85.5, the values of volume-surface mean diameter(d32) and weight mean diameter(d43) were 0.09-0.79 µm and 1.1-34.1 µm, respectively. The experimental data had significant (p < 0.05) difference with other emulsions. The value of zeta potential ranged from -34.8 to -53.1 mV, indicating that the emulsion stability of 10% OPO was the most stable in all experiment samples.

Glycidyl Fatty Acid Esters in Refined Edible Oils: A Review on Formation, Occurrence, Analysis, and Elimination Methods.

Glycidyl fatty acid esters (GEs), one of the main contaminants in processed oils, are mainly formed during the deodorization step in the refining process of edible oils and therefore occur in almost all refined edible oils. GEs are potential carcinogens, due to the fact that they readily hydrolyze into the free form glycidol in the gastrointestinal tract, which has been found to induce tumors in various rat tissues. Furthermore, glycidol has already been identified as a "possible human carcinogen'' (group 2A) by the Intl. Agency for Research on Cancer (IARC). Therefore, significant effort has been devoted to inhibit and eliminate the formation of GEs. The aim of this review is to provide a comprehensive summary on the following topics: (i) GE occurrence data for different edible oils and oil-based food products, (ii) precursors of GEs, (iii) factors influencing the formation of GEs, (iv) potential reaction mechanisms involving the leaving group and reaction intermediates, and (v) analytical methods, including the indirect and direct methods. More importantly, the various elimination methods for GEs in refined edible oils are being reviewed with focus on 3 aspects: (i) inhibition and removal of reactants, (ii) modification of reactive conditions, and (iii) elimination of GE products.

The RING finger protein NtRCP1 is involved in the floral transition in tobacco (Nicotiana tabacum).

The transition from the vegetative phase to the reproductive phase is a major developmental process in flowering plants. The underlying mechanism controlling this cellular process remains a research focus in the field of plant molecular biology. In the present work, we identified a gene encoding the C3H2C3-type RING finger protein NtRCP1 from tobacco BY-2 cells. Enzymatic analysis demonstrated that NtRCP1 is a functional E3 ubiquitin ligase. In tobacco plants, expression level of NtRCP1 was higher in the reproductive shoot apices than in the vegetative ones. NtRCP1-overexpressing plants underwent a more rapid transition from the vegetative to the reproductive phase and flowered markedly earlier than the wild-type control. Histological analysis revealed that the shoot apical meristem of NtRCP1-overexpressing plants initiated inflorescence primordia precociously compared to the wild-type plant due to accelerated cell division. Overexpression of NtRCP1 in BY-2 suspension cells promoted cell division, which was a consequence of the shortened G2 phase in the cell cycle. Together, our data suggest that NtRCP1 may act as a regulator of the phase transition, possibly through its role in cell cycle regulation, during vegetative/reproductive development in tobacco plant.

Development of genic SSR markers from transcriptome sequencing of pear buds.

A total of 8375 genic simple sequence repeat (SSR) loci were discovered from a unigene set assembled from 116282 transcriptomic unigenes in this study. Dinucleotide repeat motifs were the most common with a frequency of 65.11%, followed by trinucleotide (32.81%). A total of 4100 primer pairs were designed from the SSR loci. Of these, 343 primer pairs (repeat length ≥15 bp) were synthesized with an M13 tail and tested for stable amplification and polymorphism in four Pyrus accessions. After the preliminary test, 104 polymorphic genic SSR markers were developed; dinucleotide and trinucleotide repeats represented 97.11% (101) of these. Twenty-eight polymorphic genic SSR markers were selected randomly to further validate genetic diversity among 28 Pyrus accessions. These markers displayed a high level of polymorphism. The number of alleles at these SSR loci ranged from 2 to 17, with a mean of 9.43 alleles per locus, and the polymorphism information content (PIC) values ranged from 0.26 to 0.91. The UPGMA (unweighted pair-group method with arithmetic average) cluster analysis grouped the 28 Pyrus accessions into two groups: Oriental pears and Occidental pears, which are congruent to the traditional taxonomy, demonstrating their effectiveness in analyzing Pyrus phylogenetic relationships, enriching rare Pyrus EST-SSR resources, and confirming the potential value of a pear transcriptome database for the development of new SSR markers.

Effect of autologous platelet-rich plasma in combination with bovine porous bone mineral and bio-guide membrane on bone regeneration in mandible bicortical bony defects.

OBJECTIVE: Growth factors contained in platelet-rich plasma (PRP) can induce osteoblast differentiation in certain studies, whereas in others, osteogenesis of PRP on mandible bone defects has not been proved clinically. The aim of the study was to investigate the effect of autologous PRP on the osteogenic potential of combining bovine porous bone mineral (BPBM) and bio-guide membrane (BGM) in promoting mandible bicortical bony defects in rabbits. METHODS: One circular mandible bicortical bony defects were created in each of 54 rabbits, which were divided into 3 groups: group 1: 18 of the defects were left unfilled as a negative control; group 2: 18 of the defects were grafted with autologous PRP and BPBM/BGM; group 3: 18 of the defects were grafted with BPBM/BGM without PRP. Animals were killed at 4, 8, and 12 weeks after operation. Harvested tissue and specimens were evaluated histologically and radiographically, and metabolized observation was performed. Histological parameters associated with osteoblast activities, bone trabecula, neovascularization, newly formed mineralized bone, rudimental grafts and connective tissue formation were measured. Densities of the bones at 4, 8, and 12 weeks were studied by radiographic. The bone defect closure ratio was measured at 12 weeks. The bone metabolized parameter alkaline phosphatase was also measured and compared between 4, 8, and 12 weeks. RESULTS: The platelet concentration of PRP is 4.19- to 4.43-fold to that of the whole blood. Histological analysis showed new bone formation at all therapeutic sites including BPBM/BGM grafts with or without PRP. A statistically significant difference in new bone formation between group PRP/BPBM/BGM and group BPBM/BGM was observed. Untreated defects of group control showed the less bone regeneration. There was significant difference of bone density between group PRP/BPBM/BGM and control, and group BPBM/BGM and control, at 4, 8, and 12 weeks postoperative. There were more bone defects filling, and the grafts were absorbed at 12 weeks of group PRP/BPBM/BGM compared with group BPBM/BGM. Defects treated with PRP/BPBM/BGM demonstrated significantly increased activity of osteoblasts, enhanced amount of mitochondria and rough endoplasmic reticulum in osteoblasts, and increased concentration of alkaline phosphatase at 4, 8, and 12 weeks compared with those treated with BPBM/BGM and control group. Complete closure ratio of bone defects treated with PRP/BPBM/BGM (50%) was significantly increased compared with that treated with BPBM/BGM (16.6%). CONCLUSIONS: The study suggested that PRP combination of BPBM and BGM had significant therapeutic effects on mandible bicortical bony defects of rabbits. The effects are associated with the high concentration of platelet in PRP and the porous configuration of BPBM. Although we cannot reveal the detailed statistical relationship of PRP on promoting BPBM/GBM osteoinductive effects, PRP demonstrated superior results of bone regeneration.

Influence of heat stress on leaf ultrastructure, photosynthetic performance, and ascorbate peroxidase gene expression of two pear cultivars (Pyrus pyrifolia).

Plants encounter a variety of stresses in natural environments. One-year-old pot-grown trees of pear (Pyrus pyrifolia Nakai cv. Cuiguan and Wonhwang) were exposed to two heat stress regimes. Under constant short-term heat stress, chloroplasts and mitochondria were visibly damaged. Relative chlorophyll content and maximum photochemical efficiency of photosystem II were significantly decreased, which indicated that the leaf photosynthetic capability declined. Under chronic heat stress, mesophyll cell ultrastructure was not obviously damaged, but leaf photosynthetic capability was still restrained. As chronic heat stress was a simulation of the natural environment in summer, further study of the responses under this stress regime was undertaken. Ascorbate peroxidase (APX) activity was increased in 'Cuiguan', but not in 'Wonhwang'. Inducible expression of PpAPX genes in the cytoplasm, chloroplasts and peroxisomes was consistent with increased APX activity in 'Cuiguan', whereas only weak induction of PpAPX genes was observed in 'Wonhwang'. The isoenzymes cytosolic APX1 (cAPX1) and stromal APX (sAPX) were confirmed to be localized in the cytoplasm and chloroplasts, respectively.

Characterization and expression profiling of cucumber kinesin genes during early fruit development: revealing the roles of kinesins in exponential cell production and enlargement in cucumber fruit.

Rapid cell division and expansion in early fruit development are important phases for cucumber fruit yield and quality. Kinesin proteins are microtubule-based motors responsible for modulating cell division and enlargement. In this work, the candidate kinesin genes involved in rapid cell division and expansion during cucumber fruit development were investigated. The morphological and cellular changes during early fruit development were compared in four cucumber genotypes with varied fruit size. The correlation between the expression profiles of cucumber kinesin genes and cellular changes in fruit was investigated. Finally, the biochemical characteristics and subcellular localizations of three candidate kinesins were studied. The results clarified the morphological and cellular changes during early cucumber fruit development. This study found that CsKF2-CsKF6 were positively correlated with rapid cell production; CsKF1 and CsKF7 showed a strongly positive correlation with rapid cell expansion. The results also indicated that CsKF1 localized to the plasma membrane of fast-expanding fruit cells, that CsKF2 might play a role in fruit chloroplast division, and that CsKF3 is involved in the function or formation of phragmoplasts in fruit telophase cells. The results strongly suggest that specific fruit-enriched kinesins are specialized in their functions in rapid cell division and expansion during cucumber fruit development.

Comprehensive analysis of the transcription of starch synthesis genes and the transcription factor RSR1 in wheat (Triticum aestivum) endosperm.

The cDNA sequences of 26 starch synthesis genes were identified in common wheat (Triticum aestivum L.), and their transcript levels were measured using quantitative real-time RT-PCR to assess the function of individual genes and the regulatory mechanism in wheat endosperm. The expression patterns of 26 genes in wheat endosperm were classified into three groups. The genes in group 1 were richly expressed in the early stage of grain development and may be involved in the construction of fundamental cell machinery, synthesis of glucan primers, and initiation of starch granules. The genes in group 2 were highly expressed during the middle and late stages of grain development, and their expression profiles were similar to the accumulation rate of endosperm starch; these genes are presumed to play a crucial role in starch production. The genes in group 3 were scantily expressed throughout the grain development period and might be associated with transitory starch synthesis. Transcripts of the negative transcription factor TaRSR1 were high at the early and late stages of grain development but low during the middle stage. The expression pattern of TaRSR1 was almost opposite to those of the group 2 starch synthesis genes, indicating that TaRSR1 might negatively regulate the expression of many endosperm starch synthesis genes during grain development.

Identification of the TaBTF3 gene in wheat (Triticum aestivum L.) and the effect of its silencing on wheat chloroplast, mitochondria and mesophyll cell development.

The full-length cDNA (882bp) and DNA (1742bp) sequences encoding a basic transcription factor 3, designated as TaBTF3, were first isolated from common wheat (Triticum aestivum L.). Subcellular localization studies revealed that the TaBTF3 protein was mainly located in the cytoplasm and nucleus. In TaBTF3-silenced transgenic wheat seedlings obtained using the Virus-induced gene silencing (VIGS) method, the chlorophyll pigment content was markedly reduced. However, the malonaldehyde (MDA) and H(2)O(2) contents were enhanced, and the structure of the wheat mesophyll cell was seriously damaged. Furthermore, transcripts of the chloroplast- and mitochondrial-encoded genes were significantly reduced in TaBTF3-silenced transgenic wheat plants. These results suggest that the TaBTF3 gene might function in the development of the wheat chloroplast, mitochondria and mesophyll cell. This paper is the first report to describe the involvement of TaBTF3 in maintaining the normal plant mesophyll cell structure.

Effect of frozen storage on molecular weight, size distribution and conformation of gluten by SAXS and SEC-MALLS.

In this study, the effects of frozen (-18 °C) storage time on molecular weight, size distribution, conformation, free amino groups and free sulfhydryl groups of gluten were studied by small-angle X-ray scattering (SAXS), multi-angle laser light scattering (MALLS) in conjunction with a size exclusion chromatography (SEC) and spectrophotometrically. The results showed that the gluten dissolved in 50 mM acetic acid appeared to be similar to quasi-spherical of the chain conformation and the slope of the conformation plot decreased during the storage. Both the molecular weight and radius of gyration of the frozen gluten decreased with the storage time showing a depolymerization in the high molecular weight fraction of gluten (10(5) Da ~ 10(9) Da). Therefore, at constant molecular weight the change of the chain conformation did not show a clear correlation with the storage time. The free amino groups content changed little and the free sulfhydryl groups content of the gluten increased from 9.8 µmol/g for the control to 12.87 µmol/g for 120-day-stored gluten, indicating that the water redistribution and ice recrystallization lead to the breakage of the disulphide bonds and may be one of the reasons for the depolymerization of gluten polymer.

Arabidopsis kinesin KP1 specifically interacts with VDAC3, a mitochondrial protein, and regulates respiration during seed germination at low temperature.

The involvement of cytoskeleton-related proteins in regulating mitochondrial respiration has been revealed in mammalian cells. However, it is unclear if there is a relationship between the microtubule-based motor protein kinesin and mitochondrial respiration. In this research, we demonstrate that a plant-specific kinesin, Kinesin-like protein 1 (KP1; At KIN14 h), is involved in respiratory regulation during seed germination at a low temperature. Using in vitro biochemical methods and in vivo transgenic cell observations, we demonstrate that KP1 is able to localize to mitochondria via its tail domain (C terminus) and specifically interacts with a mitochondrial outer membrane protein, voltage-dependent anion channel 3 (VDAC3). Targeting of the KP1-tail to mitochondria is dependent on the presence of VDAC3. When grown at 4° C, KP1 dominant-negative mutants (TAILOEs) and vdac3 mutants exhibited a higher seed germination frequency. All germinating seeds of the kp1 and vdac3 mutants had increased oxygen consumption; the respiration balance between the cytochrome pathway and the alternative oxidase pathway was disrupted, and the ATP level was reduced. We conclude that the plant-specific kinesin, KP1, specifically interacts with VDAC3 on the mitochondrial outer membrane and that both KP1 and VDAC3 regulate aerobic respiration during seed germination at low temperature.

[Multiple supernumerary teeth: a case report].

Supernumerary teeth is one of the dysplasia that the number of the teeth are more than physical number. Most cases of reports were with 1-2 supernumerary teeth and rare cases were with more than 3 supernumerary teeth. A 17-year old female patient of 7 impacted supernumerary teeth were found because of toothache of premolar caused by impacted supernumerary teeth and were treated by extraction of impacted supernumerary teeth.

[Changes of IL-8 and TNF-α in gingival crevicular fluid before and after treatment from chronic periodontitis].

AIM: To investigate the changes of IL-8 and TNF-α in gingival crevicular fluid (GCF) before and after treatment from chronic periodontitis. METHODS: 96 patients with chronic periodontitis were selected as study group and experienced initial periodontal therapy. 50 healthy people were selected as control group. Gingival crevicular fluid was collected before and after treatment. At the same time, the clinical parameters, including PLI, PD, SBI and AL were recorded, and the levels of IL-8 and TNF-α in GCF were measured by ELISA in two groups. RESULTS: The levels of IL-8 and TNF-α in GCF after treatment were significantly lowered in study group as compared with those before treatment (P < 0.05), while PLI, PD, SBI and AL showed significant reduction (P < 0.05). The levels of IL-8 and TNF-α in GCF before and after treatment were significantly highered in study group as compared with those in control group (P < 0.05). There was significant correlation between PD and IL-8 in GCF (r = 0.486, P < 0.05) and between SBI, PD and TNF-α in GCF (r = 0.495, 0.648, P < 0.05). CONCLUSION: IL-8 and TNF-α may be associated with the development and the severity of periodontitis.

Recruitment of AtWHY1 and AtWHY3 by a distal element upstream of the kinesin gene AtKP1 to mediate transcriptional repression.

A 43-bp distal element, the AtKP1-related element (KPRE), was previously shown to repress the promoter activity of the kinesin gene AtKP1 in Arabidopsis thaliana. In order to identify KPRE-binding factor 1 (KBF1), a combination of ion-exchange chromatography, gel-filtration chromatography and DNA-affinity chromatography was used to purify KBF1 from whole cell extracts of Arabidopsis seedlings. Mass spectrometric identification showed that KBF1 contains two members of the whirly family of transcription factors, AtWHY1 and AtWHY3. KBF1 is a single and double-stranded DNA-binding factor. A ChIP assay showed that AtWHY1 and AtWHY3 bind to the upstream region of AtKP1 gene in vivo. Over-expression of AtWHY1 and AtWHY3 led to an obvious decrease of AtKP1 transcripts, based on quantitative real-time PCR analysis. Interestingly, salicylic acid treatment resulted in an increase of AtWHY1 and AtWHY3 transcripts, and a decrease of AtKP1 transcripts. Thus, AtWHY1 and AtWHY3, as two components of KBF1, can be recruited at the KPRE site to mediate the transcriptional repression of AtKP1. Our results prove that AtKP1 is a new downstream target of the whirly family of transcription factors.

Structural changes of malt proteins during boiling.

Changes in the physicochemical properties and structure of proteins derived from two malt varieties (Baudin and Guangmai) during wort boiling were investigated by differential scanning calorimetry, SDS-PAGE, two-dimensional electrophoresis, gel filtration chromatography and circular dichroism spectroscopy. The results showed that both protein content and amino acid composition changed only slightly during boiling, and that boiling might cause a gradual unfolding of protein structures, as indicated by the decrease in surface hydrophobicity and free sulfhydryl content and enthalpy value, as well as reduced alpha-helix contents and markedly increased random coil contents. It was also found that major component of both worts was a boiling-resistant protein with a molecular mass of 40 kDa, and that according to the two-dimensional electrophoresis and SE-HPLC analyses, a small amount of soluble aggregates might be formed via hydrophobic interactions. It was thus concluded that changes of protein structure caused by boiling that might influence beer quality are largely independent of malt variety.

[Maxillary second molar with two buccal and two lingual root canals: a case report].

Incidence rate of 4 root canals in maxillary second molar is very low and most molars have only two mesiobuccal root canals. The emergence of 4 root canals in maxillary second molar with two lingual root canals is especially rare. A case of 4 root canals maxillary second molar with two lingual root canals was successfully treated and reported in this article.

[Health values and health-related behaviors in ethnic minority groups in Guizhou Province].

OBJECTIVE: To investigate the perceived health values and its impact on health-related behaviors of the ethnic minority people in Guizhou province, with an aim to developing culturally appropriate health education and health promotion strategies. METHODS: A self-designed questionnaire survey was undertaken in 722 Miao, Gelao, Dong and Buyi people aged 15 years or over. The questionnaire contained six items evaluating perceived health values. The health-related behaviors covered smoking, alcohol drinking, safe water drinking, blood pressure check, reading health information, and seeking medical attention for illness. Kruskal-Wallis Test and Spearman correlation analysis were used to test the relationship between health values and health-related behaviors. RESULTS: Smoking and alcohol drinking were more prevalent in the minority men than in the minority women (P<0. 001). The health-related behaviors including smoking, alcohol drinking and reading changed with age and education levels. Those who had less health risk behaviors (except for smoking) tended to put more values on health. Each of the six health value items was associated with at least three health-related behaviors. Blood pressure check and reading were positively correlated with all of the six health values items. CONCLUSION: There is a need to increase perceived values of health in the ethnic minority people through education so that health promotion can be effectively implemented.

[Study on the quality of life and its influencing factors in Miao and Gelao Minority groups in Guizhou province].

OBJECTIVE: To investigate and evaluate the quality of life and its influencing factors in Miao and Gelao Minority groups in Guizhou province with the aim of providing evidence for valid health promotion measures. METHODS: Based on a self-designed questionnaire to evaluate the quality of life on 582 subjects. Kruskal-wallis test and stepwise regression analysis were used to test the quality of life and its influencing factors. RESULTS: There were significant differences among the groups of quality of life and its influencing factors. The quality of life of Miao Minority was the highest and Gelao the lowest. Sex, age, acute disease prevalence of the past two-weeks, prevalence of chronic diseases, drinking unboiled water, tobacco-smoking habit, educational level and the family expenditure on life were correlated with all or most of the domains of quality of life. CONCLUSION: The key steps on quality of life promotion should include: strengthening the basis medical health service, pushing cooperative medical service, enhancing prevention and care of chronic diseases, speeding the development of economics etc.