Lithium and an EPAC-specific inhibitor ESI-09 synergistically suppress pancreatic cancer cell proliferation and survival.

Our previous studies showed that while lithium suppresses proliferation and induces apoptosis in pancreatic cancer cells, the inhibition of exchange proteins directly activated by cyclic adenosine monophosphate (cAMP) (EPAC)1 blocks pancreatic cancer cell migration and invasion. In this study, we further investigated the combinatory effects of lithium and EPAC-specific inhibitor (ESI)-09, an EPAC-specific inhibitor, on pancreatic cancer cell proliferation and viability, and explored whether lithium synergistically cooperates with EPAC inhibition in suppressing pancreatic cancer cell tumorigenicity. The cell viability of pancreatic cancer cell lines PANC-1 and MiaPaCa-2 was measured after 48 h of incubation with different dose combination of lithium and ESI-09. Flow cytometric analysis was carried out to further verify the impact of lithium and ESI-09 upon PANC-1 cell proliferation and apoptosis. To investigate the mechanism that the effects generated by lithium and ESI-09 on PANC-1 cells, the intracellular cAMP level was measured by an ELISA-based cAMP immunoassay. Our data showed that lithium and ESI-09 synergistically inhibit pancreatic cancer cell growth and survival. Furthermore, our results revealed a novel mechanism in which the synergism between lithium and ESI-09 is not mediated by the inhibitory effect of lithium toward GSK3ß, but by lithium's ability to suppress cAMP/protein kinase A signaling.

Effects of tannic acid interfacial absorption on the physicochemical stability of algal oil-loaded emulsions and inhibition of fishy off-flavor.

In the present study, the inhibition of fishy off-flavor and destabilization in algae oil-loaded emulsions by tannic acid (TA) adsorption on octenyl succinic anhydride (OSA) starch interfaces were investigated. The changes of typical fishy off-flavor components in the emulsion, physiochemical stability, and interaction between TA and OSA starch were analyzed. The TA fortification significantly prevented the production of fishy smell-related volatile components such as heptanal and (E, E)-3, 5-octadiene-2-one. The proportion of TA on the interface was more than 90 %, forming an interfacial film with the antioxidant function. The emulsions stabilized by OSA starch-TA complexes had better oxidative and physical stability. The isothermal titration calorimetry suggested that the interaction between OSA starch and TA included hydrogen bonding and van der Waals forces (ΔG = -13.272 kJ·mol-1·K-1, ΔH = -1.302 × 103 kJ·mol-1, ΔS = -4.326 kJ·mol-1·K-1). Altogether, these results provided application guidance for developing starch-based oil-in-water emulsion systems with antioxidant properties.

The selective encapsulation and stabilization of cinnamaldehyde and eugenol in high internal phase Pickering emulsions: Regulating the interfacial properties.

This work aimed to investigate the encapsulation and stabilization mechanism of cinnamaldehyde and eugenol in high internal phase Pickering emulsions (HIPPEs) through regulating their interfacial rheological properties and interfacial microstructure. With the incorporation of cinnamaldehyde, the Schiff base reaction between the cinnamaldehyde and proteins favored the formation of the predominantly elastic and solid-like interfacial layers. In contrast, the hydrogen bonds between eugenol and proteins resulted in the transformation of interfacial layers to viscous dominant with weak viscoelastic responses. Thus, cinnamaldehyde-loaded HIPPEs had a better storage stability than eugenol-loaded HIPPEs, and the retention rate was increased by about 15 %∼20 %. The addition of tea camellia seed oil inhibited the mobility of immobilized water and improved the retention rates of cinnamaldehyde and eugenol by approximately 6 % and 12 % (30 days at 25 °C), respectively. These findings will be beneficial for the development and design of effective essential oil encapsulation systems in the food industry.

Effect of tannic acid-OSA starch complexation on the binding capacity and release of aldehydes off-flavor in aqueous matrix.

In order to further clarify the regulation of tannic acid on the off-flavor in starch-based algal oil emulsions, the effect of different starch matrix (OSA starch and OSA starch-tannic acid complex) on the release capacities of aldehydes (pentanal, hexanal, heptanal, nonanal) were investigated. The adsorption and retention ability, thermodynamic parameters, and hydrophobicity of aldehydes in the starch matrix were analyzed. Nonanal exhibited the strongest adsorption ability (65.01%-85.69%) with the starch matrix, followed by heptanal, hexanal, and pentanal, which accounted for the structures of aldehydes. Furthermore, aldehydes had a higher affinity with complex (16.33%-83.67%) than OSA starch (9.70%-66.71%) because the tannic acid altered the structure of OSA starch. Isothermal titration calorimetry suggested that the interaction between the starch matrix and aldehydes was an entropy-driven spontaneous endothermic reaction, and hydrophobic interactions were the predominant driving forces. Altogether, these results lay a theoretical foundation for facilitating the regulation of flavor in starch foods.

Changes of lipid oxidation, volatile and taste-active compounds during pan-heating of pork belly.

This study investigated the mechanisms underlying the evolution and formation of aroma and taste-active compounds of pork belly in representative traditional pork cuisines during pan-heating. The results revealed that as the temperature increased to 110 ℃, the unsaturation of fatty acids decreased from 60.25 % to 58.71 %, while the content of free radicals and secondary oxidation products increased. At the later heating stages, the addition of spices and increased heating temperature (150 â„ƒ) led to continuous increments in the contents (from 958.20 µg/kg to 1511.88 µg/kg) and diversity of volatile compounds in pork belly, imparting the unique aroma. Additionally, the accumulation of low-molecular-weight peptides, free amino acids, and nucleotides not only provided the substrate for thermal reactions and their synergistic effects, but also contributed to the desired taste quality. These findings offered insights into the flavor formation mechanisms of traditional pork cuisines and provided direction for further research.

Cytokeratin 5 positive cells represent a steroid receptor negative and therapy resistant subpopulation in luminal breast cancers.

A majority of breast cancers are estrogen receptor (ER) positive and have a luminal epithelial phenotype. However, these ER⁺ tumors often contain heterogeneous subpopulations of ER⁻ tumor cells. We previously identified a population of cytokeratin 5 (CK5) positive cells within ER⁺ and progesterone receptor positive (PR⁺) tumors that is both ER⁻PR⁻ and CD44⁺, a marker of breast tumor-initiating cells (TICs). These CK5⁺ cells have properties of TICs in luminal tumor xenografts, and we speculated that they are more resistant to chemo- and anti-ER-targeted therapies than their ER⁺ neighbors. To test this, we used ER⁺PR⁺ T47D and MCF7 breast cancer cells. CK5⁺ cells had lower proliferative indices than CK5⁻ cells, were less sensitive to 5-fluorouracil and docetaxel, and cultures became enriched for CK5⁺ cells after treatments. CK5⁺ cells were less prone to drug-induced apoptosis than CK5⁻ cells. In cells treated with 17ß-estradiol (E) plus anti-estrogens tamoxifen or fulvestrant, ER protein levels decreased, and CK5 protein levels increased, compared to controls treated with E alone. In ER⁺ tumors from patients treated with neoadjuvant endocrine therapies ER gene expression decreased, and CK5 gene expression increased in post compared to pre-treatment tumors. The number of CK5⁺ cells in tumors also increased in post- compared to pre-treatment tumors. We conclude that an ER⁻PR⁻CK5⁺ subpopulation found in many luminal tumors is resistant to standard endocrine and chemotherapies, relative to the majority ER⁺PR⁺CK5⁻ cells. Compounds that effectively target these cells are needed to improve outcome in luminal breast cancers.

Thirteen-month duration of immunity of an oral canine vaccine against challenge with Bordetella bronchiseptica.

BACKGROUND: Very few studies have evaluated the duration of immunity of Bordetella bronchiseptica vaccines in dogs, and to date, no studies have been published on the duration of immunity of oral canine Bordetella bronchiseptica vaccines. This study was designed to determine the effectiveness of a single dose of an oral B bronchiseptica vaccine in dogs when challenged 13 months after vaccination. METHODS: Two groups of approximately eight-week-old beagles were vaccinated once with 1 ml of placebo vaccine (oral, n=17) or 1 ml of Recombitek Oral Bordetella (oral, n=17). Thirteen months after vaccination, both groups were challenged with virulent B bronchiseptica via aerosolisation. RESULTS: Thirteen of 17 dogs in the placebo group (76.5 per cent) and no dogs in the Recombitek Oral Bordetella vaccine group (0.0 per cent) developed spontaneous cough of two or more consecutive days (disease case definition). Dogs in the Recombitek Oral Bordetella group had a significantly lower prevalence of disease with prevented fraction of 1 (100 per cent prevention). In addition, the number of days coughing, duration of cough and prevalence of tracheal and nasal shedding were significantly lower for dogs vaccinated with Recombitek Oral Bordetella. CONCLUSIONS: The study demonstrated that vaccination with Recombitek Oral Bordetella is effective in preventing disease and reducing shedding 13 months after vaccination when compared with dogs vaccinated with a placebo.

Lithium Suppresses Hedgehog Signaling via Promoting ITCH E3 Ligase Activity and Gli1-SUFU Interaction in PDA Cells.

Dysregulation of Hedgehog (Hh) signaling pathway is one of the hallmarks of pancreatic ductal adenocarcinoma (PDA). Lithium, a clinical mood stabilizer for the treatment of mental disorders, is known to suppress tumorigenic potential of PDA cells by targeting the Hh/Gli signaling pathway. In this study, we investigated the molecular mechanism of lithium induced down-regulation of Hh/Gli1. Our data show that lithium promotes the poly-ubiquitination and proteasome-mediated degradation of Gli1 through activating E3 ligase ITCH. Additionally, lithium enhances interaction between Gli1 and SUFU via suppressing GSK3ß, which phosphorylates SUFU and destabilizes the SUFU-Gli1 inhibitory complex. Our studies illustrate a novel mechanism by which lithium suppresses Hh signaling via simultaneously promoting ITCH-dependent Gli1 ubiquitination/degradation and SUFU-mediated Gli1 inhibition.

Accounting for dropout reason in longitudinal studies with nonignorable dropout.

Dropout is a common problem in longitudinal cohort studies and clinical trials, often raising concerns of nonignorable dropout. Selection, frailty, and mixture models have been proposed to account for potentially nonignorable missingness by relating the longitudinal outcome to time of dropout. In addition, many longitudinal studies encounter multiple types of missing data or reasons for dropout, such as loss to follow-up, disease progression, treatment modifications and death. When clinically distinct dropout reasons are present, it may be preferable to control for both dropout reason and time to gain additional clinical insights. This may be especially interesting when the dropout reason and dropout times differ by the primary exposure variable. We extend a semi-parametric varying-coefficient method for nonignorable dropout to accommodate dropout reason. We apply our method to untreated HIV-infected subjects recruited to the Acute Infection and Early Disease Research Program HIV cohort and compare longitudinal CD4+ T cell count in injection drug users to nonusers with two dropout reasons: anti-retroviral treatment initiation and loss to follow-up.

Phycocyanin prevents methylglyoxal-induced mitochondrial-dependent apoptosis in INS-1 cells by Nrf2.

Methylglyoxal (MG) is a reactive dicarbonyl compound, whose abnormal accumulation in diabetic patients exerts deleterious effects on cells and tissues. The ß-cell is the main target cell of Type 2 diabetes, and its insulin secretion injury and cell apoptosis can be due to mitochondrial dysfunction. Previous studies have demonstrated MG induced ß-cell apoptosis. However, little is known about the effect of MG on ß-cell mitochondrial dysfunction. Phycocyanin (PC) has been demonstrated to possess various biological activities including the effects on diabetic models in vivo. The aim of this study was to determine the protective effect of PC against methylglyoxal (MG)-induced dysfunction in pancreatic ß-cell INS-1 and also the mechanism. We demonstrated that MG induced mitochondrial dysfunction by the decline in ATP levels, and the increase of the level of intracellular reactive oxygen species (ROS). Furthermore, MG released cytochrome c and apoptosis-inducing factor (AIF) from the mitochondrion, induced changes in the expression of Bcl-2 family members, activated caspases and increased PARP cleavage. Interestingly, PC activated nuclear erythroid-related factor 2 (Nrf2), and Nrf2 activation as well as antioxidant enzymes HO-1 and glyoxalase 1 (Glo-1) were confirmed to be involved in the mechanisms underlying the protection of PC by RNA interference. Altogether, these results demonstrated that PC prevented mitochondrial-dependent apoptosis in MG-induced INS-1 cells and the effect was associated with Nrf2 activation.

The adenomatous polyposis coli protein is an essential regulator of radial glial polarity and construction of the cerebral cortex.

Radial glia are highly polarized cells that serve as neuronal progenitors and as scaffolds for neuronal migration during construction of the cerebral cortex. How radial glial cells establish and maintain their morphological polarity is unknown. Using conditional gene targeting in mice, we demonstrate that adenomatous polyposis coli (APC) serves an essential function in the maintenance of polarized radial glial scaffold during brain development. In the absence of APC, radial glial cells lose their polarity and responsiveness to the extracellular polarity maintenance cues, such as neuregulin-1. Elimination of APC further leads to marked instability of the radial glial microtubule cytoskeleton. The resultant changes in radial glial function and loss of APC in radial glial progeny lead to defective generation and migration of cortical neurons, severely disrupted cortical layer formation, and aberrant axonal tract development. Thus, APC is an essential regulator of radial glial polarity and is critical for the construction of cerebral cortex in mammals.

GSK-3 is a master regulator of neural progenitor homeostasis.

The development of the brain requires the exquisite coordination of progenitor proliferation and differentiation to achieve complex circuit assembly. It has been suggested that glycogen synthase kinase 3 (GSK-3) acts as an integrating molecule for multiple proliferation and differentiation signals because of its essential role in the RTK, Wnt and Shh signaling pathways. We created conditional mutations that deleted both the alpha and beta forms of GSK-3 in mouse neural progenitors. GSK-3 deletion resulted in massive hyperproliferation of neural progenitors along the entire neuraxis. Generation of both intermediate neural progenitors and postmitotic neurons was markedly suppressed. These effects were associated with the dysregulation of beta-catenin, Sonic Hedgehog, Notch and fibroblast growth factor signaling. Our results indicate that GSK-3 signaling is an essential mediator of homeostatic controls that regulate neural progenitors during mammalian brain development.