A comparative study on flaxseed lignan biotransformation through resting cell catalysis and microbial fermentation by ß-glucosidase production Lactiplantibacillus plantarum.

BACKGROUND: Flax lignan has attracted much attention because of its potential bioactivities. However, the bioavailability of secoisolariciresinol diglucoside (SDG), the main lignan in flaxseed, depends on the bioconversion by the colon bacteria. Lactic acid bacteria (LAB) with ß-glucosidase activity has found wide application in preparing bioactive aglycone. RESULTS: LAB strains with good ß-glucosidase activity were isolated from fermented tofu. Their bioconversion of flax lignan extract was investigated by resting cell catalysis and microbial fermentation, and the metabolism of SDG by Lactiplantibacillus plantarum C5 following fermentation was characterized by widely targeted metabolomics. Five L. plantarum strains producing ß-glucosidase with broad substrate specificity were isolated and identified, and they all can transform SDG into secoisolariciresinol (SECO). L. plantarum C5 resting cell reached a maximum SDG conversion of 49.19 ± 3.75%, and SECO generation of 21.49 ± 1.32% (0.215 ± 0.013 mm) at an SDG substrate concentration of 1 mM and 0.477 ± 0.003 mm SECO was produced at 4 mm within 24 h. Although sixteen flax lignan metabolites were identified following the fermentation of SDG extract by L. plantarum C5, among them, four were produced following the fermentation: SECO, demethyl-SECO, demethyl-dehydroxy-SECO and isolariciresinol. Moreover, seven lignans increased significantly. CONCLUSION: Fermentation significantly increased the profile and level of flax lignan metabolites, and the resting cell catalysis benefits from higher bioconversion efficiency and more straightforward product separation. Resting cell catalysis and microbial fermentation of flax lignan extract by the isolated ß-glucosidase production L. plantarum could be potentially applied in preparing flax lignan ingredients and fermented flaxseed. © 2024 Society of Chemical Industry.

The flax genome reveals orbitide diversity.

BACKGROUND: Ribosomally-synthesized cyclic peptides are widely found in plants and exhibit useful bioactivities for humans. The identification of cyclic peptide sequences and their precursor proteins is facilitated by the growing number of sequenced genomes. While previous research largely focused on the chemical diversity of these peptides across various species, there is little attention to a broader range of potential peptides that are not chemically identified. RESULTS: A pioneering study was initiated to explore the genetic diversity of linusorbs, a group of cyclic peptides uniquely occurring in cultivated flax (Linum usitatissimum). Phylogenetic analysis clustered the 5 known linusorb precursor proteins into two clades and one singleton. Preliminary tBLASTn search of the published flax genome using the whole protein sequence as query could only retrieve its homologues within the same clade. This limitation was overcome using a profile-based mining strategy. After genome reannotation, a hidden Markov Model (HMM)-based approach identified 58 repeats homologous to the linusorb-embedded repeats in 8 novel proteins, implying that they share common ancestry with the linusorb-embedded repeats. Subsequently, we developed a customized profile composed of a random linusorb-like domain (LLD) flanked by 5 conserved sites and used it for string search of the proteome, which extracted 281 LLD-containing repeats (LLDRs) in 25 proteins. Comparative analysis of different repeat categories suggested that the 5 conserved flanking sites among the non-homologous repeats have undergone convergent evolution driven by functional selection. CONCLUSIONS: The profile-based mining approach is suitable for analyzing repetitive sequences. The 25 LLDR proteins identified herein represent the potential diversity of cyclic peptides within the flax genome and lay a foundation for further studies on the functions and evolution of these protein tandem repeats.

Castor patatin-like phospholipase A IIIß facilitates removal of hydroxy fatty acids from phosphatidylcholine in transgenic Arabidopsis seeds.

KEY MESSAGE: Castor patatin-like phospholipase A IIIß facilitates the exclusion of hydroxy fatty acids from phosphatidylcholine in developing transgenic Arabidopsis seeds. Hydroxy fatty acids (HFAs) are industrial useful, but their major natural source castor contains toxic components. Although expressing a castor OLEATE 12-HYDROXYLASE in Arabidopsis thaliana leads to the synthesis of HFAs in seeds, a high proportion of the HFAs are retained in phosphatidylcholine (PC). Thus, the liberation of HFA from PC seems to be critical for obtaining HFA-enriched seed oils. Plant phospholipase A (PLA) catalyzes the hydrolysis of PC to release fatty acyl chains that can be subsequently channeled into triacylglycerol (TAG) synthesis or other metabolic pathways. To further our knowledge regarding the function of PLAs from HFA-producing plant species, two class III patatin-like PLA cDNAs (pPLAIIIß or pPLAIIIδ) from castor or Physaria fendleri were overexpressed in a transgenic line of A. thaliana producing C18-HFA, respectively. Only the overexpression of RcpPLAIIIß resulted in a significant reduction in seed HFA content with concomitant changes in fatty acid composition. Reductions in HFA content occurred in both PC and TAG indicating that HFAs released from PC were not incorporated into TAG. These results suggest that RcpPLAIIIß may catalyze the removal of HFAs from PC in the developing seeds synthesizing these unusual fatty acids.

The linin promoter is highly effective in enhancing punicic acid production in Arabidopsis.

KEY MESSAGE: Enhanced levels of punicic acid were produced in the seed oil of Arabidopsis over-expressing pomegranate FATTY ACID CONJUGASE driven by heterologous promoters, among which the linin promoter was the most efficient. Fatty acids with conjugated double bonds play a special role in determining both the nutritional and industrial uses of plant oils. Punicic acid (18:3Δ9cis,11trans,13cis ), a conjugated fatty acid naturally enriched in the pomegranate (Punica granatum) seeds, has gained increasing attention from the biotechnology community toward its production in metabolically engineered oilseed crops because of its significant health benefits. The present study focused on selecting the best heterologous promoter to drive the expression of the P. granatum FATTY ACID CONJUGASE (PgFADX) cDNA as a means of producing punicic acid in Arabidopsis seed oil. Among the four promoters of genes encoding seed storage proteins from different crop species, the linin promoter led to the highest accumulation of punicic acid (13.2% of total fatty acids in the best homozygous line). Analysis of the relative expression level of PgFADX in developing seeds further confirmed that the linin promoter was most efficient in Arabidopsis. In addition, a conserved profile of cis-regulatory elements were identified in four heterologous promoters by bioinformatic analysis, and their possible roles in regulating gene expression during plant development were also discussed based on the results of this study in combination with the literature. This study contributes to metabolic engineering strategies aimed at enhancing the production of bioactive fatty acids in oilseed crops.

Bioconversion of lignans in flaxseed cake by fermented tofu microbiota and isolation of Enterococcus faecium strain ZB26 responsible for converting secoisolariciresinol diglucoside to enterodiol.

Human intestinal microbiota plays a crucial role in converting secoisolariciresinol diglucoside, a lignan found in flaxseed, to enterodiol, which has a range of health benefits: antioxidative, antitumor, and estrogenic/anti-estrogenic effects. Given the high secoisolariciresinol diglucoside content in flaxseed cake, this study investigated the potential of co-fermenting flaxseed cake with fermented soybean product to isolate bacterial strains that effectively convert secoisolariciresinol diglucoside to enterodiol in a controlled environment (in vitro). The co-fermentation process with stinky tofu microbiota significantly altered the lignan, generating 12 intermediate lignan metabolites as identified by targeted metabolomics. One particular promising strain, ZB26, demonstrated an impressive ability to convert secoisolariciresinol diglucoside. It achieved a conversion rate of 87.42 ± 0.33%, with secoisolariciresinol and enterodiol generation rates of 94.22 ± 0.51% and 2.91 ± 0.03%, respectively. Further optimization revealed, under specific conditions (0.5 mM secoisolariciresinol diglucoside, pH 8, 30 °C for 3 days), ZB26 could convert an even higher percentage (97.75 ± 0.05%) of the secoisolariciresinol diglucoside to generate secoisolariciresinol (103.02 ± 0.16%) and enterodiol (3.18 ± 0.31%). These findings suggest that the identified strains ZB26 have promising potential for developing functional foods and ingredients enriched with lignans.

Stability of tryptophan-containing LOs in flaxseed oil and their response towards γ-tocopherol.

Linusorbs (LOs), significantly influence oil quality and sensory properties of flaxseed oil. Trp-containing LOs exhibit distinct oxidative behavior when γ-tocopherol (γ-T) is present. Polar fractions of crude flaxseed oil were stripped via silica absorption, and reintroduced (LO and γ-T) separately into the oil matrix to investigate their interaction during storage. Compared with crude oil, LOs account for 18.49% reduction of p-anisidine value, while LOs with γ-T contributed to most of the endogenous antioxidant effect in crude oil. γ-T was found to suppress oxidation of Trp-containing LO at early stage (Met form), while facilitate oxidation while at their mid-stage (MetO form, Methionine sulfoxide). In vitro oxidation shows that CLD more likely cleaved into peptide fragments, while few products retain intact ring structures. LC-MS/MS analysis and silicon simulation revealed proximity between MetO and Trp residues, facilitating inter- or intra-molecular reactions and ring structure rupture. Remarkably, the presence of γ-T facilitate these phenomena.

A two-step method of cyclolinopeptide (linusorb) preparation from flaxseed cake via dry-screening.

Linusorbs (LO), cyclolinopeptides, are a group of cyclic hydrophobic peptides and considered a valuable by-product of flaxseed oil due to numerous health benefits. Currently applied acetone or methanol extraction could contaminate the feedstocks for further food-grade application. Using flaxseed cake as feedstock, this study established a practical method for preparing LO from pressed cake. Firstly, LO composition of 15 flaxseed cultivars was analyzed. Next, cold-pressed cake was milled and screened mechanically. The kernel and hull fractions were separated based on the disparity of their mechanical strength. Monitored by hyperspectral fluorescence, the LO-enriched kernel fraction separated from cold-pressed flaxseed cake was further used as feedstock for LO production. After ethanol extraction, partition, and precipitation, LOs were extracted from cold-pressed flaxseed cake with a purity of 91.4%. The proposed method could serve as feasible flaxseed cake valorization strategy and enable the preparation of other polar compounds such as flax lignan and mucilage.

Temporal and Spatial Changes of Proarrhythmic Substrate in Premature Ventricular Contraction-Induced Cardiomyopathy.

BACKGROUND: The changes in proarrhythmic substrates and malignant ventricular arrhythmia mechanisms caused by premature ventricular contraction-induced cardiomyopathy (PVCCM) remain unclear. OBJECTIVES: The goal of this study was to establish the electrophysiological mechanism of how high-load PVC causes malignant arrhythmia. METHODS: Thirteen swine were exposed to 50% paced PVC from the right ventricular apex for 12 weeks (PVCCM, n = 6) and no pacing for 12 weeks (control, n = 7). Cardiac function was quantified biweekly with echocardiography. Computed tomography scans and electrophysiological examinations were performed monthly to dynamically evaluate the changes in the cardiac structure and the arrhythmogenic substrate. RESULTS: The decreases in the cardiac function and ventricular enlargement in the PVCCM group were significant after 12 weeks of PVC stimulation compared with the control group (P < 0.001). Electrophysiological examination found that the ventricular effective refractory period dispersion (0.071 ± 0.008), area of the low-voltage zone (9.41 ± 1.55 cm2), and malignant ventricular arrhythmia inducibility (33.3%) of the PVCCM group increased significantly at week 8 after pacing (P < 0.001 vs the control group); these changes slowed down after 8 weeks. Moreover, the distribution of the low-voltage zone presented obvious spatial heterogeneity, especially in the anterior wall of the right ventricle, accompanied by delayed activation in the sinus rhythm (67 ± 13 milliseconds). Consistently, the proportion of ventricular fibrosis- and expression-related proteins were significantly increased in the PVCCM group (P < 0.001), especially in the right ventricle. Moreover, proteomic analysis confirmed the spatial profile of these fibrotic changes in the PVCCM group. CONCLUSIONS: High-burden PVC can cause significant temporal and spatial heterogeneity changes in proarrhythmic substrates, which are potentially related to the upregulation of calcium signaling caused by asynchronous activation.

Right atrial appendage: an important structure to drive atrial fibrillation.

PURPOSE: Understanding of the atrial fibrillation (AF) driven by right atrial appendage (RAA) is limited. This study aimed to understand the characteristics of the AF driven by RAA and explore ablation methods. METHODS: This was a retrospective study and patients who were identified as having the AF driven by RAA were reviewed. Ablation was performed during AF. Potential maps of the left and right atrium, electrophysiological examinations, and ablation methods were studied. RESULTS: Among the 20 identified patients (mean age 67.0 ± 11.2 years; ejection fraction 62.9 ± 6.0%; LA diameter 43.1 ± 4.9 mm; RA diameter 51.7 ± 8.3 × 42.9 ± 3.7 mm), the AF cycle length in RAA (134.0 ± 10.9 ms) was the shortest, and the fastest frequency potentials were located in the RAA in 65% of patients. For the left atrium, the AF cycle length of the roof (145.5 ± 14.9 ms) was the shortest, followed by the left atrial appendage (153.7 ± 17.1 ms) and bottom (154.8 ± 11.8 ms). High-frequency potentials of RAA could be rapidly conducted to left atrium via sagittal bundle and Bachmann's bundle, and the conduction time (55.0 ± 5.0 ms) was significantly shorter than the mean bi-atrial activation time (176.7 ± 10.3 ms, P < 0.0001). AF could be terminated after ablation at the RAA base (17 patients) or mechanical stimulation within the RAA (3 patients). To date, only two patients had recurrent atrial flutter, while the remaining patients maintained sinus rhythm. CONCLUSION: The AF driven by RAA is characterized by high-frequency potentials in RAA, and ablation at the RAA base can achieve a satisfactory therapeutic effect.

Absence of Rgs5 Influences the Spatial and Temporal Fluctuation of Cardiac Repolarization in Mice.

AIMS: This study investigated the contribution of the regulator of G-protein signaling 5 (Rgs5) knockout to the alteration of the action potential duration (APD) restitution and repolarizing dispersion in ventricle. METHODS AND RESULTS: The effects of Rgs5-/- were investigated by QT variance (QTv) and heart rate variability analysis of Rgs5-/- mice. Monophasic action potential analysis was investigated in isolated Rgs5-/- heart. Rgs5-/- did not promote ventricular remodeling. The 24-h QTv and QT variability index (QTVI) of the Rgs5-/- mice were higher than those of wild-type (WT) mice (P < 0.01). In WT mice, a positive correlation was found between QTv and the standard deviation of all NN intervals (r = 0.62; P < 0.01), but not in Rgs5-/- mice (R = 0.01; P > 0.05). The absence of Rgs5 resulted in a significant prolongation of effective refractory period and APD in isolated ventricle. In addition, compared with WT mice, the knockout of Rgs5 significantly deepened the slope of the APD recovery curve at all 10 sites of the heart (P < 0.01) and increased the spatial dispersions of Smax (COV-Smax) (WT: 0.28 ± 0.03, Rgs5-/-: 0.53 ± 0.08, P < 0.01). Compared with WT heart, Rgs5-/- increased the induced S1-S2 interval at all sites of heart and widened the window of vulnerability of ventricular tachyarrhythmia (P < 0.05). CONCLUSION: Our findings indicate that Rgs5-/- is an important regulator of ventricular tachyarrhythmia in mice by prolonging ventricular repolarization and increasing spatial dispersion in ventricle.

Clinical Safety and Efficacy of Ablation for Atrial Fibrillation Patients With a History of Stroke.

Objectives: To evaluate the clinical safety and efficacy of radiofrequency catheter ablation for atrial fibrillation patients with a history of stroke. Methods and Results: A total of 116 symptomatic, drug-refractory AF patients with a history of stroke, and 1:2 matched patients without a history of stroke were enrolled. Of these, 28 cases occurred stroke within 3 months (Group 1), 88 cases with stroke history longer than 3 months (Group 2), and 232 cases without stroke (Group 3). PVI was performed in all patients, extended to ablation of linear lesions ablation. The periprocedural stroke rates and other procedure-related in-hospital complications did not differ significantly among the three groups. The maintenance rate of SR after the procedure showed no significant difference (p = 0.333), 52.7, 66.4, and 70.7% in Group 1, 2, and 3, respectively. Furthermore, the comparison between a history of stroke and those without it were also shown no significant difference (p = 0.351). Conclusions: Radiofrequency ablation for AF patients occurred stroke, even within 3 months is safe and effective, without higher periprocedural complication rate and recurrence rate.

Self-adaptive cardiac optogenetics device based on negative stretching-resistive strain sensor.

Precision medicine calls for high demand of continuous, closed-loop physiological monitoring and accurate control, especially for cardiovascular diseases. Cardiac optogenetics is promising for its superiority of cell selectivity and high time-space accuracy, but the efficacy of optogenetics relative to the input of light stimulus is detected and controlled separately by discrete instruments in vitro, which suffers from time retardation, energy consumption, and poor portability. Thus, a highly integrated system based on implantable sensors combining closed-loop self-monitoring with simultaneous treatment is highly desired. Here, we report a self-adaptive cardiac optogenetics system based on an original negative stretching-resistive strain sensor array for closed-loop heart rate recording and self-adaptive light intensity control. The strain sensor exhibits a dual and synchronous capability of precise monitor and physiological-electrical-optical regulation. In an in vivo ventricular tachycardia model, our system demonstrates the potential of a negative stretching-resistive device in controlling-in-sensor electronics for wearable/implantable autodiagnosis and telehealth applications.

A Study of Cardiogenic Stroke Risk in Non-valvular Atrial Fibrillation Patients.

Objectives: We attempted to develop more precisely quantified risk models for predicting cardiogenic stroke risk in non-valvular atrial fibrillation (NVAF) patients. Methods: We conducted a case-control study, using data from hospitalized patients with AF who underwent transesophageal echocardiography at Shanghai Chest Hospital. A total of 233 high cardiogenic stroke risk patients with left atrial appendage thrombus (LAT) or left atrial spontaneous echo contrast (LA-SEC) and 233 controls matched for age, sex, AF type. Results: AF history, LA diameter enlargement, larger left ventricular end diastolic diameter, lower ejection fraction, greater serum uric acid (SUA), and brain natriuretic peptide (BNP) levels showed association with high stroke risk. The multivariate logistic regression analysis revealed that AF duration, left atrial diameter (LAd), left ventricular ejection fraction (LVEF), SUA, and BNP were independent risk factors of the LAT/LA-SEC. We used LAd, LVEF, SUA, and BNP to construct a combined predictive model for high stroke risk in NVAF patients (the area under ROC curve: 0.784; sensitivity 66.1%; specificity 76.8%; 95% CI 0.744-0.825, P < 0.001). Conclusion: Comprehensive evaluation of LAd, LVEF, SUA, and BNP may help stratify the cardiogenic stroke risk among non-valvular AF patients, guiding anticoagulation therapy.

Localized Controlled Delivery of Gemcitabine via Microsol Electrospun Fibers to Prevent Pancreatic Cancer Recurrence.

The low radical surgery rate of pancreatic cancer leads to increased local recurrence and poor prognosis. Gemcitabine (GEM) is the preferred chemotherapeutic for pancreatic cancer. However, systemic chemotherapy with GEM has reached a bottleneck due to its serious side effects after frequent injections. In this study, GEM is successfully enwrapped into electrospun fibers via microsol electrospinning technology to form a stable core-shell fibrous structure. The GEM release rate can be adjusted by altering the thickness of the hyaluronan-sol inner fiber and the quantity of loaded GEM, and the release can be sustained for as long as three weeks. In vitro assays show that these electrospun fibers effectively inhibit pancreatic cancer cells and promote apoptosis. In vivo studies show that the fibrous membranes are better for inhibiting the growth of residual tumors than that of integrated tumors. Furthermore, immunohistochemistry results show that GEM-loaded fibers promote a higher cell apoptosis rate than does systemically injected GEM in residual tumors. In addition, the local delivery of GEM with fibers significantly reduces liver toxicity. In summary, a core-shell electrospun fiber for the controlled and localized delivery of GEM, which greatly improves the treatment of residual tumors and prevents pancreatic tumor recurrence, is developed.

Gambogic acid sensitizes gemcitabine efficacy in pancreatic cancer by reducing the expression of ribonucleotide reductase subunit-M2 (RRM2).

BACKGROUND: Pancreatic cancer is susceptible to gemcitabine resistance, and patients receive less benefit from gemcitabine chemotherapy. Previous studies report that gambogic acid possesses antineoplastic properties; however, to our knowledge, there have been no specific studies on its effects in pancreatic cancer. Therefore, the purpose of this study was to explore whether increases the sensitivity of pancreatic cancer to gemcitabine, and determine the synergistic effects of gambogic acid and gemcitabine against pancreatic cancer. METHODS: The effects of gambogic acid on cell viability, the cell cycle, and apoptosis were assessed using 4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT) and flow cytometry in pancreatic cancer cell lines. Protein expression was detected by western blot analysis and mRNA expression was detected using q-PCR. A xenograft tumor model of pancreatic cancer was used to investigate the synergistic effects of gambogic acid and gemcitabine. RESULTS: Gambogic acid effectively inhibited the growth of pancreatic cancer cell lines by inducing S-phase cell cycle arrest and apoptosis. Synergistic activity of gambogic acid combined with gemcitabine was observed in PANC-1 and BxPC-3 cells based on the results of MTT, colony formation, and apoptosis assays. Western blot results demonstrated that gambogic acid sensitized gemcitabine-induced apoptosis by enhancing the expression of cleaved caspase-3, cleaved caspase-9, cleaved-PARP, and Bax, and reducing the expression of Bcl-2. In particular, gambogic acid reduced the expression of the ribonucleotide reductase subunit-M2 (RRM2) protein and mRNA, a trend that correlated with resistance to gemcitabine through inhibition of the extracellular signal-regulated kinase (ERK)/E2F1 signaling pathway. Treatment with gambogic acid and gemcitabine significantly repressed tumor growth in the xenograft pancreatic cancer model. Immunohistochemistry results demonstrated a downregulation of p-ERK, E2F1, and RRM2 in mice receiving gambogic acid treatment and combination treatment. CONCLUSIONS: These results demonstrate that gambogic acid sensitizes pancreatic cancer cells to gemcitabine in vitro and in vivo by inhibiting the activation of the ERK/E2F1/RRM2 signaling pathway. The results also indicate that gambogic acid treatment combined with gemcitabine might be a promising chemotherapy strategy for pancreatic cancer.

In vitro and in vivo combined antibacterial effect of levofloxacin/silver co-loaded electrospun fibrous membranes.

Post-surgery mesh infections are one of the most common complications of hernia repair at the interface of implant materials and tissue, because high doses of antimicrobial agents are toxic and low doses of antibacterial agents are ineffective, with no good clinical solution currently available. To reduce the infection rates after mesh implantation, we designed a "one-pot synthesized" mesoporous silica nanoplatform consisting of levofloxacin (Lev) and silver (Lev@MSN@Ag) composites with poly-l-lactide (PLLA) electrospun fibrous membranes via blending electrospinning. With advances in the combined antibacterial agents Lev and Ag at a low dosage for the treatment of drug-resistant bacterial infections (28 µg mL-1 Lev and 12 µg mL-1 Ag) with a concentration of 5 × 105 CFU mL-1, the composite electrospun fibers act as a carrier for drug-loading and have an antibacterial effect over 8 weeks. Lev@MSN@Ag-PLLA fibers showed a superior antibacterial effect on drug resistant strains in the in vitro test at low doses of antibacterial agents. Further, the in vivo study showed that Lev@MSN@Ag-PLLA fibers significantly inhibited bacterial growth and infection over 8 weeks through the combined effect of low dosage antibacterial drugs. In conclusion, the Lev@MSN@Ag-PLLA fibers provided an advanced combined antibacterial nanoplatform of low dosage for the treatment of drug-resistant bacterial infections.

Production of Diacylglycerol-enriched Oil by Glycerolysis of Soybean Oil using a Bubble Column Reactor in a Solvent-free System.

In this study, diacylglycerol-enriched soybean oil (DESO) was synthesized through Lipozyme 435-catalyzed glycerolysis of soybean oil (SO) in a solvent-free system using a modified bubble column reactor. The effects of enzyme load, mole ratio of glycerol to soybean oil, reaction temperature, gas flow and reaction time on DAG production were investigated. The selected conditions were established as being enzyme load of 4 wt% (mass of substrates), glycerol/soybean oil mole ratio of 20:1, reaction temperature of 80°C, gas flow of 10.6 cm/min, and a reaction time of 2.5 h, obtaining the DAG content of 49.4±0.5 wt%. The reusability of Lipozyme 435 was evaluated by monitoring the contents of DAG, monoacylglycerol (MAG) and triacylglycerol (TAG) in 10 consecutive runs. After purified by one-step molecular distillation, the DAG content of 63.5±0.3 wt% was achieved in DESO. The mole ratio of 1, 3-DAG to 1, 2-DAG was 2:1 and the fatty acid composition had no significant difference from that of soybean oil. However, the thermal properties of DESO and SO had considerable differences. Polymorphic form of DESO were mainly the ß form and minor amounts of the ß' form. Granular aggregation and round-shaped crystals were detected in DESO.

Overexpression of NDC80 is correlated with prognosis of pancreatic cancer and regulates cell proliferation.

AIMS: NDC80/Hec1, one of four proteins of the outer kinetochore NDC80 complex, is involved in the tumorigenesis of a variety of cancers. In this study, we focused on that NDC80 is overexpressed in human pancreatic cancer and investigates the role of NDC80-knockdown in pancreatic cancer cells proliferation. MATERIALS AND METHODS: We determined the expression levels of NDC80 on both mRNA and protein levels in fresh pancreatic cancer tissues and cells by quantitative real-time polymerase chain reaction and immunoblotting, respectively. Furthermore, protein level of NDC80 was identified using immunochemistry in paraffin-embedded tumor specimen, with correlation between NDC80 expression and various clinicopathological parameters evaluated. The role of NDC80 in pancreatic cancer cells (Panc-1) growth was investigated by lentivirus-mediated silencing of NDC80. The effect of NDC80 deletion on cell proliferation was analyzed by MTT assay and clone formation assay, while cell cycle distributions and apoptosis were analyzed by flow cytometry. RESULTS: The mRNA and protein of NDC80 were overexpressed in pancreatic cancer tissues and cells. The statistical analysis based on immunohistochemical evaluation suggested that NDC80 overexpression was signifi cantly associated with clinicopathological parameters including pathological T staging and N staging, which may be served as an predictor for poor outcomes. The silencing of NDC80 in Panc-1 cells could suppress cell proliferation and colony formation. Furthermore, the NDC80-siRNA infected Panc-1 cells lead to cell cycle arrest at G2/M phase and induction of apoptosis. CONCLUSION: These results demonstrated that NDC80 plays an essential role in the tumorigenesis of pancreatic cancer, and might serve as potential prognostic and therapeutic target for treatment of pancreatic cancer.

Structure and protective effect on UVB-induced keratinocyte damage of fructan from white garlic.

The fructose polymer fructan was extracted from white garlic and fractionated using DEAE cellulose 52 and Sephadex G-100 columns to characterize its chemical composition and protective effect against ultraviolet radiation b (UVB) induced human keratinocyte (HaTaC) damage. Gel permeation chromatography, high performance anion exchange chromatography, infrared spectroscopy and 1D and 2D nuclear magnetic resonance spectroscopy were used to determine the chemical composition and functional characteristics of the garlic fructan (GF). GF was a homogeneous polysaccharide with a molecular weight of 4.54 × 10(3)Da. It was a member of the 1-kestose family, and it was composed of fructose and glucose at a ratio of 14:1. The main chain of GF was composed of (2→1)-ß-D-fructopyranose linked to a terminal (2→1)-α-D-glucopyranose at the non-reducing end and a (2→6)-ß-D-fructopyranose branched chain. The degree of polymerization was 28. Preliminary tests described herein indicated that GF may be effective in protecting HaTaC from UVB-induced damage.