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The continued improvement of genetics, nutrition, and management has resulted in rapid growth, better feed efficiency, and higher meat yield with competitive prices in the broiler industry. Nowadays, however, it is well-documented that productive traits and fertility are negatively correlated, and male broiler breeders are exposed to a fertility decline after 45 wk of age. Considering a low male-to-female ratio in breeder flocks, roosters have a prominent impact on flock fertility. Consequently, strategies to maintain the fertility of male broiler breeders could guarantee the reproductive performance of commercial herds. Understanding reproductive aging demands deep insights into its molecular and physiological mechanisms. Over-weighting, Sertoli and Leydig cell dysfunctions, compromised antioxidant capacity, imbalance in sexual hormones, and epididymal lithiasis are among candidate culprits associated with reproductive aging in roosters. Nutritional and managing strategies have been successfully applied to modulate body weight, improve sperm fatty acid profile and antioxidant status, and boost spermatogenic and steroidogenic pathways. The current review characterizes the physiology and biochemistry of reproductive aging in male broiler breeders and then highlights strategies and their underlying mechanisms to mitigate this failure. In summary, applying one or more of the abovementioned strategies might result in consistent post-peak reproduction and benefit producers in the poultry industry. [ABSTRACT FROM AUTHOR]

DEPTOR plays vital roles in the regulation of cell proliferation and survival by directly modulating the activity of mTORC1/2. However, the physiological role of DEPTOR in lung tumorigenesis, as well as its clinical significance, remains elusive. In this study, we revealed that decreased DEPTOR expression correlated with increased tumor size, poor differentiation, and worse survival in patients with lung cancer. DEPTOR depletion promoted cell proliferation, survival, migration, and invasion in human lung cancer cells. Mechanistically, DEPTOR bound to the kinase domain of EGFR via its PDZ domain to inactivate EGFR signal. Thus, DEPTOR depletion not only directly activated mTORC1/2, but also relieved the inhibition of EGFR to subsequently activate mTOR signals, leading to the induction of cell proliferation and survival. Additionally, activated EGFR-mTOR signals upregulated the expression of ZEB1 and SLUG to induce epithelial-mesenchymal transition, resulting in enhanced migration and invasion. Importantly, Deptor deletion accelerated KrasG12D;p53fl/fl-induced lung tumorigenesis and shortened mouse life span via the activation of EGFR-mTOR signals. Collectively, our study demonstrated that DEPTOR acts as a tumor suppressor in lung tumorigenesis, and its reduction may advance the progression of human lung cancer. [ABSTRACT FROM AUTHOR]

Effective treatment regimens for triple-negative breast cancer (TNBC) are relatively scarce due to a lack of specific therapeutic targets. Epidermal growth factor receptor (EGFR) signaling is highly active in TNBC and is associated with poor prognosis. Most EGFR antagonists, which significantly improve outcome in lung and colon cancer, have shown limited clinical effects in breast cancer. However, limiting EGFR expression in TNBC is a potential strategy for improving the clinical efficacy of EGFR antagonists. Here, we found that the gamma-aminobutyric acid type A receptor π subunit (GABRP), as a membrane protein enriched in TNBC stem cells, interacted with EGFR and significantly sustained its expression, resulting in stemness maintenance and chemotherapy resistance. Silencing GABRP induced down-regulation of EGFR signaling, which hindered cell stemness and enhanced sensitivity to chemotherapies, including paclitaxel, doxorubicin, and cisplatin. We also identified that retigabine, an FDA-approved drug for adjunctive treatment of seizures, increased the sensitivity of EGFR to gefitinib in gefitinib-resistant cells. Our findings show that GABRP can sustain the stemness of TNBC via modulating EGFR expression, suggesting that GABRP may be a potential therapeutic target that can address EGFR inhibitor resistance in TNBC. [ABSTRACT FROM AUTHOR]

The physiology and biochemistry of young Opuntia spp. cladodes relate with their Crassulacean acid metabolism, which extends over the day-night cycle in four phases, is species-dependent and is affected by water availability. This study aimed to assess the interaction among species, time-of-day, and the soil water potential (Ψ W) on biochemical and physiological characteristics of cladodes of Opuntia species. Three-week-old cladodes were harvested at 7:00 a.m. and 3:00 p.m. from plants with or without irrigation for 30 d (−0.17 and −5.72 MPa soil Ψ W), from O. albicarpa , O. ficus-indica , O. hyptiacantha , O. megacantha , and O. streptacantha. The experimental design was a factorial 5 x 2 x 2 (species, sampling time and soil Ψ W). The experimental unit was one cladode per plant, and six repetitions were evaluated. Total acids, glucose, fructose, sucrose, starch, total phenolics, free amino acids, and soluble proteins concentrations were evaluated, as well as acid invertase and neutral invertase activities. The interaction among species x soil Ψ W and species x time of the day was significant (P ≤ 0.05) in all variables evaluated. An exception was the species x soil Ψ W on starch concentration (P = 0.1827). The biochemical and physiological characteristics of Opuntia cladodes were modified by the time of the day and soil Ψ W interaction, but most of the characteristics were positively or inversely affected depending on the species, frequently displaying a descending trend following O. streptacantha , O. hyptiacantha , O. megacantha , O. albicarpa and O. ficus-indica. The total acids, glucose, fructose, starch, soluble proteins, and free amino acids concentrations revealed that domestication significantly modifies C and N metabolism in Opuntia. • The complex domestication process is only incipiently known and has been studied in a small subset of crops. • Extreme environment stimulates physiological and biochemical characteristics in the genus Opuntia related to domestication. • Physiology and biochemistry of Opuntia cladodes depend on the interaction of species, domestication, CAM phase, and watering. • Total acids, sugars, proteins, and free amino acids revealed that domestication modify C and N metabolism in genus Opuntia. [ABSTRACT FROM AUTHOR]

The experiment was implemented to assess the influence of dietary supplementation of laying quails with creatine monohydrate (CrM), L-carnitine (CAR) and their mixture (CrMCAR) as antioxidants against oxidative stress (OS) induced by 2.5 ppm lead acetate (LA) in drinking water on productive, physiological and microbial aspects. In total, 400 laying quail females at 10 wk of age were divided into a randomized design with 5 groups and 4 replicates of 20 birds each. Birds were fed ad libitum with a balanced diet for 8 wk. The control group was kept under no-stress conditions and was given fresh water without any additives (G1). While birds in other groups were exposed to OS induced experimentally by 2.5 ppm LA in drinking water with no feed additive (G2) or supplemented with 500 mg/kg CrM (G3) or 500 mg/kg CAR (G4) or combination of 250 mg/kg each of CrM and CAR (CrMCAR, G5) to feed mixture. Compared to G2, G5 demonstrated the reduction (P ≤ 0.05) of feed conversion ratio, feed intake, mortality and ileal total coliform, as well as serum and egg malondialdehyde and serum lipid hydroperoxide, uric acid, glucose, cholesterol, enzymatic activities (alanine aminotransferase, aspartate transaminase, alkaline phosphatase, creatine phosphokinase, γ-glutamyl transferase), and heterophils/lymphocytes ratio. In the meanwhile, there was an increase (P ≤ 0.05) in egg production, egg mass, and weight with the improvement of egg quality, serum sex hormones level and ileal lactic acid bacteria for G5 followed by G4 and G3. Moreover, G5 enhanced (P ≤ 0.05), the total antioxidant capacity of egg and serum glutathione, superoxide dismutase, catalase, glutathione peroxidase, protein and calcium levels. Therefore, dietary CrMCAR, CAR and CrM have analogous influence to control by improving the antioxidant and physiological parameters which resulted in better productive performance and egg characteristics of stressed quails. These antioxidants, especially in their equal combination, are beneficial to alleviate oxidative stress incidence and can be recommended for poultry feeding under various aspects of environmental stresses. [ABSTRACT FROM AUTHOR]

Understanding of the range and mechanisms of clathrin functions has developed exponentially since clathrin's discovery in 1975. Here, newly established molecular mechanisms that regulate clathrin activity and connect clathrin pathways to differentiation, disease and physiological processes such as glucose metabolism are reviewed. Diversity and commonalities of clathrin pathways across the tree of life reveal species-specific differences enabling functional plasticity in both membrane traffic and cytokinesis. New structural information on clathrin coat formation and cargo interactions emphasises the interplay between clathrin, adaptor proteins, lipids and cargo, and how this interplay regulates quality control of clathrin's function and is compromised in infection and neurological disease. Roles for balancing clathrin-mediated cargo transport are defined in stem cell development and additional disease states. [ABSTRACT FROM AUTHOR]

Background: Probiotic Lactobacillius rhamnosus GG (LGG) shows beneficial immunomodulation on cultured cell lines in vitro and in mouse models.Objective: The aim was to investigate the effects of LGG on intestinal injury and the underlying mechanisms by elucidating inflammatory signaling pathways and metabolomic response to LPS stimulation in the piglet intestine.Methods: Piglets (Duroc × Landrace × Large White, including males and female; 8.6 ± 1.1 kg) aged 28 d were assigned to 3 groups (n = 6/group): oral inoculation with PBS for 2 wk before intraperitoneal injection of physiological saline [control (CON)] or LPS (25 μg/kg body weight; LPS) or oral inoculation with LGG for 2 wk before intraperitoneal injection of LPS (LGG+LPS). Piglets were killed 4 h after LPS injection. Systemic inflammation, intestinal integrity, inflammation signals, and metabolomic characteristics in the intestine were determined.Results: Compared with CON, LPS stimulation significantly decreased ileal zonula occludens 1 (ZO-1; 44%), claudin-3 (44%), and occludin (41%) expression; increased serum diamineoxidase (73%), D-xylose (19%), TNF-α (43%), and IL-6 (55%) concentrations; induced p38 mitogen-activated protein kinase (p38 MAPK; 85%), extracellular signal-regulated kinase (ERK; 96%), and NF-κB p65 phosphorylation (37%) (P < 0.05). Compared with LPS stimulation alone, LGG pretreatment significantly enhanced the intestinal barrier by upregulating expressions of tight junction proteins (ZO-1, 73%; claudin-3, 55%; occludin, 67%), thereby decreasing serum diamineoxidase (26%) and D-xylose (28%) concentrations, and also reduced serum TNF-α expression (16%) and ileal p38 MAPK (79%), ERK (43%) and NF-κB p65 (37%) phosphorylation levels (P < 0.05). Metabolomic analysis showed clear separation between each group. The concentrations of caprylic acid [fold-change (FC) = 2.39], 1-mono-olein (FC = 2.68), erythritol (FC = 4.62), and ethanolamine (FC = 4.47) significantly increased in the intestine of LGG + LPS piglets compared with the LPS group (P < 0.05).Conclusions: These data suggest that LGG alleviates gut inflammation, improves intestinal barrier function, and modulates the metabolite profile of piglets challenged with LPS. This trial was registered at the Zhejiang University (http://www.lac.zju.edu.cn) as ZJU20170529. [ABSTRACT FROM AUTHOR]

Kainate receptors belong to the ionotropic glutamate receptor family and play critical roles in the regulation of synaptic networks. The kainate receptor subunit GluK3 has unique functional properties and contributes to presynaptic facilitation at the hippocampal mossy fiber synapses along with roles at the post-synapses. To gain structural insights into the unique functional properties and dynamics of GluK3 receptor, we imaged them via electron microscopy in the apo-state and in complex with either agonist kainate or antagonist UBP301. Our analysis of all the GluK3 full-length structures not only provides insights into the receptor transitions between desensitized and closed states but also reveals a "non-classical" conformation of neurotransmitter binding domain in the closed-state distinct from that observed in AMPA and other kainate receptor structures. We show by molecular dynamics simulations that Asp759 influences the stability of the LBD dimers and hence could be responsible for the observed conformational variability and dynamics of the GluK3 via electron microscopy. Lower dimer stability could explain faster desensitization and low agonist sensitivity of GluK3. In overview, our work helps to associate biochemistry and physiology of GluK3 receptors with their structural biology and offers structural insights into the unique functional properties of these atypical receptors. [ABSTRACT FROM AUTHOR]

Autophagy is an evolutionarily conserved process whereby damaged and redundant components of the cell are degraded in structures called autophagolysosomes. Currently, three main types of autophagy are recognized: macroautophagy, microautophagy, and chaperone-mediated autophagy (CMA). However, we still know little about some specific types of autophagy that are linked to various intracellular compartments and their roles in the physiology of the whole organism and connections to various diseases. Here, we aim to shed light on the latest insights on and mechanisms of several selective forms of autophagy. The evolutionarily conserved process for the elimination of damaged and redundant components in cells is known as autophagy. Currently three forms of autophagy are described: macroautophagy, microautophagy, and chaperone-mediated autophagy. Recent advances in biochemistry and cell biology have revealed the selectivity of autophagy toward the various intracellular compartments. These selective types of autophagy are essential to ensure normal functioning of the cell. Dysregulation of autophagy and its selective types can cause numerous diseases, including neurodegeneration, microbial infections, and cancer. Understanding the molecular mechanisms of selective types of autophagy will help in the development of new compounds and therapeutic strategies to fight a wide spectrum of diseases. [ABSTRACT FROM AUTHOR]

This study evaluated the quality of minimally processed mangoes irradiated with six different fluences (0, 1.5, 3.0, 4.2, 5.7 and 7.2 × 104 J m−2) of nonthermal pulsed light (PL). On the 1st, 7th and 14th days of storage at 5 °C, the mango cubes were evaluated for several quality indicators and components of physical products, such as color (L * and b *), electrolyte leakage (EL), titratable acidity (TA), pH, soluble solids (SS), vitamin C, pectinamethylesterase and polygalacturonase activity, histological evaluations, antioxidant enzyme activities, and firmness severely. Independent dose fluency treatment, it was observed that PL did not change enzymatic antioxidant system and firmness of fresh-cut mango, keeping its cell wall integrity, which could be verified by optical microscopy. On the other hand, the intensity of the applied PL exhibited different effects for the postharvest quality maintenance of fresh cut mango, evidenced also by principal component analysis (PCA). The lower fluences of PL (1.5 and 3.0 × 104 J m−2) did not affect the color (L * and b *), membrane permeability (indicated by EL), vitamin C content and the SS/TA ratio, suggesting maintenance of the maturity of fresh-cut mango. Meanwhile, higher fluences (5.7 and 7.2 × 104 J m−2) promoted a slight loss in the vitamin C content. Some variables associated with color, such as b* and C, showed decreased values during refrigerated storage. These results suggest that the application of lower fluences of PL exhibited the best results for fresh-cut mango postharvest. [ABSTRACT FROM AUTHOR]

Marine bivalves are found as key components of coastal habitats and have several important roles, such as serving as a food source for human beings and aquatic organisms. In fact, as the world's population continues to grow, bivalve aquaculture is expected to increase in importance as a means of addressing demands for animal protein; however, its growth may be possibly compromised by unfavourable climatic conditions. Thus, we assessed the effects of increased water temperature and acidification on the seeds of a bivalve of commercial importance, the Manila clam, Ruditapes philippinarum , in order to understand how this species may be affected by climate change at its early life stages. We examined the expected response of clams by experimentally mimicking seasonal conditions that could be forecasted to occur at the end of the twenty-first century. Different physiological responses were measured including growth rates, clearance rate, burrowing time and different biochemical biomarkers of metabolic stress. The results showed that growth decreased in acidic experimental conditions in spring, with a weak interaction with temperature. Clearance rate was negatively affected by a lower pH in spring and summer but, under extreme summer conditions, the effect of pH was overridden by the negative impact of a higher temperature. Burrowing rates were reduced by half under warm temperature conditions in spring and summer. In contrast, biochemical biomarkers were only significantly depicted under climate change conditions in autumn. Overall, this study demonstrates the need to consider seasonality when evaluating the potential effects of climate change on clam aquaculture in order to forecast consequences for biological production. • Growth, feeding, burrowing and biomarkers respond to Climate Change (CC) in clams. • Spring and summer are the seasons when clams are more affected by CC. • In clams, pH induced more biochemical/physiological alterations than temperature. • Seasonality is an important factor to modulate physiological responses to CC in clams. [ABSTRACT FROM AUTHOR]

Microplastics (MPs) and per- and polyfluoroalkyl substances (PFASs) have drawn significant attention as emerging threats to aquatic ecosystems. There are currently just a few investigations on the combined toxicity of PFAS and MP on freshwater microalgae. In this research, the combined toxicity of polyvinyl chloride (PVC) and perfluorooctanoic acid (PFOA) to Microcystis aeruginosa was investigated. The results indicated that the combination of these pollutants inhibited the growth of M. aeruginosa and promoted the synthesis and release of Microcystin-LR (MC-LR). Individual and combined exposure caused different responses to cellular oxidative stress. Under the Individual exposure of PFOA, when the concentration was greater than 20.0 mg/L, the catalase (CAT) activity increased significantly, and when it was greater than 100.0 mg/L, the malondialdehyde (MDA) content increased significantly, but there is no significant change under combined exposure. PVC and PFOA exposure also caused physical damage to the algal cells and reduced the content of extracellular polymer substances (EPS) based on analysis of cell morphology. Metabolic analysis revealed that carbohydrate metabolism and amino acid metabolism of the algae were affected. The current study offers a fresh theoretical framework for MPs and PFASs environmental risk evaluations. [Display omitted] • PVC and PFOA significantly inhibit algal growth. • Combined exposure promoted the synthesis and release of Microcystin-LR. • PVC and PFOA triggered oxidative stress and cytomembrane damage. • Metabolomics was used to revealed the effect of PVC and PFOA on M. aeruginosa. [ABSTRACT FROM AUTHOR]

Background: Dysregulation of glucocorticoid receptors has been implicated in addiction and stress-related disorders. FKBP5 is a co-chaperone of the glucocorticoid receptor and regulates receptor sensitivity. While FKBP5 is known to be involved in mood- and stress-related disorders, less is known regarding FKBP5 and cocaine abuse. This study investigated the regulation of FKBP5 expression in the extended amygdala and paraventricular nucleus of the hypothalamus, regions important in the control of stress-responses and HPA axis function, following chronic and acute cocaine administration.Methods: Adult male and female rats received saline or cocaine three times per day for 1 or 14 days. Brain tissue was collected 30 min, 24 h, 48 h, 7 days or 14 days following the final injection. FKBP5 mRNA was measured by qRT-PCR in the central nucleus of the amygdala (CeA), bed nucleus of the stria terminalis (BNST) and paraventricular nucleus (PVN).Results: FKBP5 mRNA levels were significantly elevated as a result of chronic cocaine administration in both males and females in the PVN and BNST 30 min and 24 h after the final injection. In females, FKBP5 was also elevated in the CeA. Following acute cocaine, FKBP5 gene expression was unaltered except for elevated levels in the BNST of females 24 h later.Conclusions: These results demonstrate that FKBP5 mRNA is regulated by cocaine administration. Increased FKBP5 expression may play a role in the dysregulation of the stress axis following chronic cocaine exposure, contributing to the negative affective symptoms of cocaine withdrawal. [ABSTRACT FROM AUTHOR]

Upregulation of special AT-rich sequence-binding protein-1 (SATB1) has been suggested to promote tumor growth and metastasis. However, the factors governing its cellular levels remain unclear. Here, we report that ubiquitin-specific peptidase 47 (USP47), a member of the deubiquitinating enzymes family, interacts with SATB1 and mediates its deubiquitination and stability. USP47 deficiency impairs transcriptional activity of SATB1 target genes and inhibits colon cancer cell proliferation, migration, and tumorigenesis in a mouse model of colon cancer. Furthermore, we identified SMURF2 as an E3 ubiquitin ligase that promotes SATB1 degradation by upregulating its ubiquitination, and its deficiency promotes colon cancer cell proliferation and SATB1 target gene transcription. SMURF2 is negatively regulated by USP47, and USP47 depletion sensitizes colon cancer cells to 5-FU treatment-induced apoptosis. Taken together, our findings provide a ubiquitination-related mechanistic link to USP47, SMURF2, and SATB1 and suggest that USP47 might be targeted for colon cancer treatment when SATB1 is overexpressed. [ABSTRACT FROM AUTHOR]

Background: Long non-coding RNAs (lncRNAs) have established roles in the pathogenesis of diverse human disorders including neuropsychiatric disorders.Methods: In the current study, we evaluated expression levels of six apoptosis-related lncRNAs (CCAT2, TUG1, PANDA, NEAT1, FAS-AS1 and OIP5-AS1) in the peripheral blood of bipolar disorder (BD) patients and healthy subjects to assess their contribution in the pathogenesis of BD.Results: CCAT2, TUG1 and PANDA were up-regulated in total BD patients compared with total healthy subjects (P values = 0.006, <0.001 and 0.004 respectively) while OIP5-AS1 was down-regulated (P = 0.001). When expression levels of these genes were compared between patients and sex-matched healthy subjects, CCAT2 and TUG1 expression levels were only different in male subgroups; while PANDA expression was different in both male and female subgroups compared with the corresponding control subgroups. Transcript levels of lncRNAs were not correlated with any of demographic or clinical parameters of BD patients or controls after adjustment for gender. Pairwise correlations between expression levels of lncRNAs followed a disease-dependent manner. Based on receiver operating characteristic curves, among the assessed lncRNAs TUG1 had the highest diagnostic power in BD. Combination of transcript levels of CCAT2, TUG1, PANDA and OIP5-AS1 improved both sensitivity and specificity resulting in diagnostic power of 0.96.Conclusion: Our data demonstrated a possible role of certain lncRNAs in the pathogenesis of BD and potentiated them as diagnostic markers in this disorder. [ABSTRACT FROM AUTHOR]

Abstract Crustaceans are exposed to a range of environmental factors that can impact their condition, physiological function and growth. Condition indices are broadly defined as the extent to which stored nutrient reserves allow normal physiological function and growth, but can also represent more than nutrition alone. There is currently no reliable indicator to non-destructively measure shrimp physiological or nutritional condition. In this study, haemolymph and biochemical indices were benchmarked against a range of physiological status condition indicators, including haemolymph biochemistry parameters and carcass traits, in fed and unfed sub-adult black tiger shrimp (Penaeus monodon) and across the moult cycle. Results based on fluorescent marker injection and analysis of recovered amounts showed that haemolymph volume was elevated immediately after the moult, but decreased significantly within 2 days and remained stable for the remainder of the inter-moult period. Brix index (Brix) strongly correlated with haemolymph biochemical, shrimp condition and carcass composition indices. These included haemolymph volume, total protein and triglycerides, as well as gross energy, hepatosomatic index (HSI) and body weight gain per moult. Overall, results demonstrated that Brix was the simplest, fastest and most cost-effective indicator to accurately and non-destructively estimate physiological condition in P. monodon. Given the correlations with a broad range of indices, Brix may represent a more holistic estimate of condition, incorporating multifactorial aspects of shrimp condition including moult cycle and nutrition. Additionally, the baseline measurements of metabolites across the moult and under starvation conditions improves our fundamental understanding of overall condition in P. monodon. Highlights • Haemolymph biochemistry correlated with shrimp condition and carcass composition. • BRIX successfully classified the nutritional condition in 90.7% of P. monodon. • We report a simple, fast, accurate and non-destructive estimate of shrimp condition. • Refractive index successfully classified the nutritional condition in P. monodon. [ABSTRACT FROM AUTHOR]

Background: The recent NIH mandate to consider sex as a biological variable in preclinical research has focused attention on delineation of sex differences in behavior. To investigate mechanisms underlying sex differences in Δ9-tetrahydrocannabinol (THC) effects, we examined the effects of sex and gonadal hormones on CB1 receptors in cerebellum, hippocampus, prefrontal cortex, and striatum.Methods: Adult Sprague-Dawley rats underwent gonadectomy (GDX) or sham-GDX. Half of the GDX females and males received estradiol or testosterone replacement (GDX+H), respectively. All rats were injected with vehicle or 30 mg/kg THC twice daily for 1 week before brain collection. CP55,940-stimulated [35S]GTPγS and [3H]SR141716A saturation binding assays were performed.Results: With exception of enhanced receptor activation in the hippocampi of female rats compared to males, vehicle-treated rats exhibited minimal sex differences in CB1 receptor densities or G-protein coupling. Repeated treatment with THC resulted in pronounced CB1 receptor desensitization and downregulation in both sexes in all brain regions with a greater magnitude of change in females.Conclusions: These results suggest that sex differences in the density and G-protein coupling of brain CB1 receptors may play a limited role in sex differences in acute THC effects not mediated by the hippocampus. In contrast, sex differences after repeated THC were common, with females (intact, GDX, and GDX+H) showing greater downregulation or desensitization in all four brain regions compared to the respective male groups. This result is consistent with a finding that women tend to progress to tolerance and dependence quicker than men after initiation of cannabis use. [ABSTRACT FROM AUTHOR]

Background: Neurotransmitter substance P (SP) and its preferred neurokinin-1 receptor (NK1R) have been implicated in the treatment of affective and addiction disorders. Despite promising preclinical data on antidepressant action, the clinical trials of NK1R antagonists in major depression have been disappointing. There are no direct in vivo imaging studies on NK1R characteristics in patients with a major depressive disorder (MDD).Methods: In this cross-sectional case-control study, we recruited nine never-medicated patients with moderate to severe MDD and nine matched healthy controls. NK1R availability (NK1R binding potential, BPND) was measured with in vivo 3-D positron emission tomography and a specific NK1 receptor tracer [18F]SPA-RQ. Clinical symptoms were assessed with the 17-item Hamilton Rating Scale for Depression (HAM-D17).Results: NK1R-BPND did not differ statistically significantly between patients with MDD and healthy controls. HAM-D17 total scores (range 21-32) correlated positively with NK1R-BPND in cortical and limbic areas. HAM-D17 subscale score for anxiety symptoms correlated positively with NK1R-BPND in specific brain areas implicated in fear and anxiety.Limitations: Small sample size. Low variability in the clinical HAM-D subscale ratings may affect the observed correlations.Conclusions: Our preliminary results do not support a different baseline expression of NK1Rs in a representative sample of never-medicated patients with MDD during a current moderate/severe depressive episode. The modulatory effect of NK1Rs on affective symptoms is in line with early positive results on antidepressant action of NK1 antagonists. However, the effect is likely to be too weak for treatment of MDD with NK1R antagonists alone in clinical practice. [ABSTRACT FROM AUTHOR]

Abstract Background The aim of the study was to assess the degree of liver damage in a rabbit perfused with histidine-tryptophan-ketoglutarate (HTK [Custodiol]) solution with and without the presence of prolactin (PRL) based on biochemical studies in perfundate and ultrastructural analysis of hepatocytes. Materials and Methods The experiment was carried out on rabbits. Liver ischemia was used in the study, based on Pringle's maneuver. About 70% of the rabbit liver lobes were perfused with HTK with or without the addition of PRL (2.5μg/g liver/h) under ischemic conditions for 2 hours. In the perfundate, the activity of enzymes alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), γ-glutamyl transpeptidase (GGT), and lactate concentration were determined. Liver biopsies were collected for histopathologic evaluation under an electron microscope. Results The addition of PRL to the HTK significantly reduced the leakage of enzymes from the liver to perfundate compared with the control group without PRL. The activity of ALT, AST, LDH, and GGT in the perfundates obtained after 2-hour perfusion with HTK-PRL solution was lower when compared with activity of the same parameters determined in perfundates with liver perfused with HTK without PRL. The area under the curve (AUC 0-2h) calculated for GGT, LDH, and lactates was significantly higher after perfusion with the HTK than with HTK with the addition of PRL. In the study group, bile was secreted throughout the whole experiment. The morphological confirmation of these results was obtained by means of transmission microscopy. Conclusion PRL added to the preservation solution significantly inhibits the process of liver cell cytolysis, which may suggest its hepatoprotective effect. Highlights • Biochemical results indicate delay of liver injuries in the prolactin group. • There was a decrease in number of hepatocytes showing necrotic damage in prolactin group. • In the study group, bile was secreted throughout the whole experiment. [ABSTRACT FROM AUTHOR]

With rapid improvements in next-generation sequencing technologies, our knowledge about metabolism of many organisms is rapidly increasing. However, gaps in metabolic networks exist due to incomplete knowledge (e.g., missing reactions, unknown pathways, unannotated and misannotated genes, promiscuous enzymes, and underground metabolic pathways). In this review, we discuss recent advances in gap-filling algorithms based on genome-scale metabolic models and the importance of both high-throughput experiments and detailed biochemical characterization, which work in concert with in silico methods, to allow a more accurate and comprehensive understanding of metabolism. [ABSTRACT FROM AUTHOR]

Objectives: Prescription stimulants are vulnerable to oral and parenteral abuse. Intravenous forms of abuse may be most detrimental due to an enhanced risk of dependence, overdose, and infectious diseases. Our objective was to discover an orally active prodrug of a stimulant that would not be easily converted to its parent when injected, thus hindering intravenous abuse.Methods: Following an initial analysis of stimulant structures, the fencamfamine isomer [(-)-FCF; (N-ethyl-3-phenylbicyclo[2.2.1]heptan-2-amine)] was chosen as a parent drug due to its favorable biochemical properties. Subsequently, PRX-P4-003 {(-)-N-(Octadecanoyloxymethoxycarbonyl)-N-ethyl-3-phenylbicyclo[2.2.1]heptan-2-amine} qualified for further development. Experimental testing of PRX-P4-003 included radioligand binding assays, stability studies, and rodent pharmacokinetic and locomotor assays.Results: Prodrug PRX-P4-003 is a pharmacologically inactive, hydrophobic compound, whereas its parent (-)-FCF is a dopamine reuptake inhibitor with weaker effects on norepinephrine reuptake (Ki = 0.07 and 0.80 μM, respectively). PRX-P4-003 is metabolized to (-)-FCF in simulated intestinal fluid (with pancreatin) but not in simulated gastric fluid (with pepsin). Finally, PRX-P4-003 shows a significant oral but no intravenous increase in locomotion, correlating with its pharmacokinetics by these different routes of administration.Conclusions: PRX-P4-003 is a novel prodrug stimulant enzymatically activated in the gut. Our data suggest a pancreatic, lipase-based mechanism of activation and as only 1% of this enzyme is found in the systemic circulation, PRX-P4-003 is unlikely to be bioactive if injected intravenously. Enzymatic release of (-)-FCF is needed prior to its systemic absorption, which may discourage oral abuse (e.g., by chewing). PRX-P4-003 is being developed for apathy in Alzheimer's disease and binge eating disorder. [ABSTRACT FROM AUTHOR]

Background: A key component of alcohol dependence (AD), a severe form of alcohol use disorder, is the negative emotional state during withdrawal. Monoamine oxidase A (MAO-A) is an important enzyme that metabolizes monoamines and creates oxidative stress. Elevations in MAO-A level, especially in the prefrontal and anterior cingulate cortex (PFC and ACC), are associated with low mood states, including the dysphoria of early alcohol withdrawal in humans. The aim of the present study was to determine whether chronic alcohol vapor exposure causes an upregulation of MAO-A activity or level in the PFC and ACC of rodents during acute withdrawal.Methods: Sprague-Dawley rats were exposed to ethanol vapor or control condition for 17 h per day for 8 weeks. MAO-A activity and protein levels were measured immediately after exposure, acute withdrawal (24 h), protracted withdrawal (4 day), and protracted abstinence (3 weeks) (n = 16/group; 8 alcohol exposed, 8 control).Results: Chronic ethanol vapor exposure significantly elevated MAO-A activity and protein levels in the PFC and ACC at 24-h withdrawal (multivariate analysis of variance (MANOVA), activity: F2,13 = 3.82, p = .05, protein: F2,13 = 5.13, p = .02). There were no significant changes in MAO-A level or activity at other timepoints.Conclusions: The results of this study suggest a causal relationship between acute alcohol withdrawal and elevated MAO-A levels and activity, clarifying the observation of greater MAO-A binding in human alcohol withdrawal. This has important implications for developing methods of targeting MAO-A and/or sequelae of its dysregulation in alcohol dependence. [ABSTRACT FROM AUTHOR]

Drought is the most important crop production limiting factor in the changing climate scenario and its intensity is predicted to increase in future. Sunflower is an important oilseed crop having 8% share in the world oilseed production. Although, it is a moderately drought tolerant crop, severe drought causes reduction in the seed and oil production. Therefore, to ensure sustainable sunflower achene and oil production, it is very important to understand the relationship among the physiological, biochemical, genetic and agronomic basis of drought for its sustainable management. Impact of drought stress on various aspects of sunflower has been reported earlier but there is not a single study describing the physiological, biochemical and genetic basis of drought in sunflower at molecular and crop level. In this review manuscript, influence of drought on sunflower achene yield and oil quality has been analyzed critically at both cell, plant and crop level, and the possible management options to mitigate the severity of the drought stress are proposed. Available literature describing the impact of drought stress on physiological and biochemical aspects (like, photosynthesis, water relations, nutrient uptake and oxidative damage), morphological and growth parameters and achene yield and oil quality has been discussed critically. Based on the discussion on the impact of drought stress, various management strategies, such as breeding for drought tolerance (conventional or biotechnological), exogenous application of hormones and osmoprotectants, seed treatment and soil nutrient management has been reviewed and discussed. It is concluded from discussion that sunflower responds to water stress by osmotic adjustments, turgor maintenance, carbon assimilation maintenance and hormonal regulations. A comprehensive research on integration of different management options, including agronomic management, conventional breeding and modern biotechnological advances, is needed for the sustainable improvement of sunflower achene yield and oil quality under drought stress. This may also contribute significantly under a climate change scenario. [ABSTRACT FROM AUTHOR]

Purpose: To determine the role of macrophage polarization on the response of inflammatory breast cancer (IBC) cells to radiation and whether modulation of macrophage plasticity can alter radiation response.Methods and Materials: The human THP-1 monocyte cell line and primary human monocytes isolated from peripheral blood mononuclear cells were differentiated into macrophages and polarized to either an "antitumor" (M1) or a "protumor" (M2) phenotype. These polarized macrophages were co-cultured with IBC cells (SUM149, KPL4, MDA-IBC3, or SUM190) without direct contact for 24 hours, then subjected to irradiation (0, 2, 4, or 6 Gy). Interleukin (IL)4/IL13-induced activation of STAT6 signaling was measured by Western blotting of phospho-STAT6 (Tyr641), and expression of M2 polarization gene markers (CD206, fibronectin, and CCL22) was measured by quantitative polymerase chain reaction.Results: Expression of M2 polarization markers was higher in M2-polarized macrophages after IL4/IL13 treatment than in control (M0) or M1-polarized macrophages. Co-culture of IBC cell lines with M1-polarized THP-1 macrophages mediated radiosensitivity of IBC cells, whereas co-culture with M2-polarized macrophages mediated radioresistance. Phosphopeptide mimetic PM37, targeting the SH2 domain of STAT6, prevented and reversed IL4/IL13-mediated STAT6 phosphorylation (Tyr641) and decreased the expression of M2 polarization markers. Pretreatment of M2-THP1 macrophages with PM37 reduced the radioresistance they induced in IBC cells after co-culture. Targeted proteomics analysis of IBC KPL4 cells using a kinase antibody array revealed induction of protein kinase C zeta (PRKCZ) in these cells only after co-culture with M2-THP1 macrophages, which was prevented by PM37 pretreatment. KPL4 cells with stable short hairpin RNA knockdown of PRKCZ exhibited lower radioresistance after M2-THP1 co-culture.Conclusions: These data suggest that inhibition of M2 polarization of macrophages by PM37 can prevent radioresistance of IBC by down-regulating PRKCZ. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the precise mechanism of recognition and processing of ionizing radiation (IR)-induced complex DNA damage (CDD), where two or more DNA lesions are in close proximity, in cellular DNA which is packaged with histones to form chromatin.Methods and Materials: HeLa and oropharyngeal squamous cell carcinoma (UMSCC74A and UMSCC6) cells were irradiated with high linear energy transfer (LET) α-particles or protons, versus low-LET protons and X rays. At various time points after irradiation, site-specific histone post-translational modifications were analyzed by quantitative Western blotting; DNA damage and repair were measured by different versions of the comet assay; and cell survival was determined using clonogenic assays.Results: Site-specific histone post-translational modifications after low- and high-LET radiation, particularly proton irradiation, were screened, aiming to identify those responsive to CDD. We demonstrate that histone H2B ubiquitylated on lysine 120 (H2Bub) is specifically induced several hours after irradiation in response to high-LET α-particles and protons but not by low-LET protons or X rays/γ-radiation. This is associated with increased levels of CDD, which contributes to decreased cell survival. We further discovered that modulation of H2Bub is under the control of two E3 ubiquitin ligases, MSL2 and RNF20/RNF40 complex, whose depletion leads to defective processing and further persistence of CDD, and to additional decreased cell survival after irradiation.Conclusion: This study demonstrates that the signaling and repair of CDD, particularly induced by high-LET IR is co-ordinated through the specific induction of H2Bub catalyzed by MSL2 and RNF20/40, a mechanism that contributes significantly to cell survival after irradiation. [ABSTRACT FROM AUTHOR]

Invertase or β-fructofuranosidase (EC 3.2.1.26) belongs to the glycoside hydrolase family 32, which catalyzes the hydrolysis of sucrose into fructose and glucose. Here, we report the biochemical and functional characterization of invertase Bmsuc1 from Bombyx mori . Bmsuc1 showed optimal hydrolysis at pH 7.0–8.0 and its optimum temperature is 50 °C using sucrose as substrate. Circular dichroism spectra indicated Bmsuc1 had a primarily β-strand structure. The thermal denaturations transition of Bmsuc1 was a cooperative process with a Tm, ΔH , and ΔS of 53.81 ± 0.12 °C, 185.51 ± 0.14 KJ/mol and 0.56 ± 0.01 KJ/(mol K), respectively. Moreover, homology modeling and multi-sequence alignment suggested that Bmsuc1 has a canonical β-propeller fold and one conserved catalytic triad, Asp 63 -Asp 181 -Glu 234 , which is located in the bottom of the substrate-binding pocket. Bmsuc1 was expressed at high levels in the silk gland at both the transcriptional and translational levels. These expression profiles combined with invertase activity analyses of Bmsuc1 suggested that it might function as a digestive enzyme to hydrolyze sugar in the silk gland lumen. Collectively, these findings expand towards a better understanding of the structure of Bmsuc1 and its function in the silk gland. [ABSTRACT FROM AUTHOR]

Harmonized methodologies are urgently required for the taste evaluation of novel pediatric medicines. This study utilized in vitro , in vivo and clinical data to evaluate the palatability of a novel midazolam chocolate tablet. In vitro dissolution experiments showed the crushed tablet to release within 5 min 1.68 mg of midazolam into simulated saliva. This translated to a drug level of 0.84 mg/ml in the oral cavity, which would be higher than the midazolam bitterness detection threshold concentration of 0.03 mg/ml determined in a rat ‘brief access taste aversion’ (BATA) model. The visual analogue scale scores of patients aged 4–16 years prescribed with midazolam pre-surgery showed a clear preference for the midazolam chocolate tablets (3.35 ± 1.04, n  = 20) compared to the control midazolam solution (1.47 ± 0.62, n  = 17). The clinical data was in agreement with the in vivo rodent data in showing the novel chocolate tablet matrix to be effective at taste-masking the bitter midazolam. [ABSTRACT FROM AUTHOR]

[6]-Gingerol, one of the components of the rhizome of Ginger, has a variety of biological activities such as anticoagulant, antioxidative, antitumor, anti-inflammatory, antihypertensive, and so forth. However, as one of the homologous phenolic ketones, [6]-gingerol is insoluble in water which limits its applications. Herein, we prepared [6]-gingerol proliposomes through modified thin-film dispersion method, which was spherical or oval, and physicochemically stable with narrow size distribution. Surprisingly, in vitro release of [6]-gingerol loaded proliposome compared with the free [6]-gingerol was significantly higher and its oral bioavailability increased 5-fold in vivo. Intriguingly, its antitumor effect was enhanced in the liposome formulation. Thus, our prepared [6]-gingerol proliposome proved to be a novel formulation for [6]-gingerol, which significantly improved its antitumor effect. [ABSTRACT FROM AUTHOR]

Background: Recent studies found abnormal biochemical metabolism and executive cognitive deficits in acute bipolar disorder (BD). However, the evidence concerning in euthymic BD is limited. Thus, a comparison between acute and euthymic BD is conductive to better understanding the association between cognition and the outcome of neuroimaging. This study sought to investigate the relationship between the executive function and the biochemical metabolism in acute- and euthymic-episode BD patients and delineate the prominent endophenotype of BD.Methods: Three groups of participants were recruited in this study: 30 BD patients with an acute depressive episode, 22 euthymic BD patients, and 31 healthy controls. All participants were interviewed using the Structured Clinical Interview for DSM-IV, and underwent two-dimensional multivoxel proton magnetic resonance spectroscopy (1H-MRS) to obtain the bilateral metabolite levels in the lenticular nucleus of basal ganglia(BG). The ratios of N-acetylaspartate (NAA)/creatine (Cr) and Choline-containing compounds (Cho) /Cr ratios were calculated. Executive function was assessed by using the Wisconsin Card Sorting Test (WCST) and Trail Making Test, Part-B(TMT-B).Results: The comparison of biochemical changes showed that the NAA/Cr ratios in bilateral lenticular nucleus in both acute and euthymic BD patients was significantly lower than that in healthy controls at a confidence level of p<0.05. In the comparison of executive function, both acute and euthymic BD patients showed significantly decreased numbers of categories completed, and increased numbers of total errors, perseverative and noperseverative errors, and TMT-B uptake compared to the healthy controls at a confidence level of p<0.05. There were no significant differences between the acute BD and euthymic BD groups in the biochemical metabolite ratios and executive function. We found that the NAA/Cr ratio in the left in BG in the acute -episode BD patients was positively correlated with the number of categories completed, whereas it was negatively correlated with the total errors and TMT-B uptake. There was no correlation between the NAA/Cr and Cho/Cr ratios in the bilateral BG and the scores of SWCT and TMT-B in euthymic-episode BD patients.Limitation: The sample size was relatively small and not all the euthymic-episode patients are the ones with an acute episode.Conclusions: Our findings suggest that biochemical abnormalities in the lenticular nucleus and the executive dysfunction may occur early in the course of BD, and persist during remission, and are the most likely markers of endophenotypes of BD. The dysfunction of the neuronal function in the lenticular nucleus may be correlated with the cold dysfunction in patients with acute BD. [ABSTRACT FROM AUTHOR]

The objective of this study was to compare the laying performance, egg quality, and serum biochemistry of hens maintained in conventional cage rearing system (CRS), flat net-rearing system (NRS), and free range system (FRS) under summer conditions. Indigenous Xianju chickens (n = 540) were randomly allocated into cages or pens of rearing system groups, within each system there were 5 replicates with 36 hens in each replicate. The experiment lasted between 21 and 29 wk of age. Hen-day egg production (P = 0.00) and egg mass (P = 0.00) were higher in the CRS but were similar in the NRS and FRS. Lowest egg weight (P = 0.02), yolk weight (P = 0.00) and yolk ratio (P = 0.01), and feed intake (P = 0.01) were observed from the FRS, whereas lowest feed conversion ratio (FCR) was recorded from the CRS (P = 0.01). Rearing systems had negligible effect on egg quality. Serum Ca (P = 0.04) and total protein (P = 0.03) levels were found to be higher in the CRS but were lower in the FRS. Serum levels of glucose (P = 0.01), cholesterol (P = 0.00), and triglyceride (P = 0.00) in the CRS increased compared with the NRS and FRS groups, whereas serum levels of high density lipoprotein cholesterol (HDL-C; P = 0.01) in the CRS decreased. It can be concluded that under summer conditions, Xianju chickens from CRS had an advantage in terms of productivity parameters, but exhibited higher levels of serum lipids and glucose. [ABSTRACT FROM AUTHOR]

Iron (Fe) deficiency is one of the most common problems of soybean. It causes upper leaves yellowing, interveinal chlorosis, stunted growth and yield loss. Manganese (Mn) deficiency affects the reactions in the oxygen evolving complex (OEC) of photosystem II and increase the accumulation of reactive oxygen species (ROS). The aim of this research is to study the effect of Fe and Mn deficiencies applied separately and simultaneously on physiological, biochemical, nutritional and growth (morphological) parameters of soybean cultivars (Glycine max L.). The experiment was conducted in nutrient hydroponic solution lacking Fe or Mn or both Fe and Mn. Chlorophyll content index (CCI) and chlorophyll a fluorescence were measured through time to detect nutritional disorders at an early growth stage before the apparition of visual symptoms. The results showed that Fe and Mn deficiencies had a significant negative effect on the photosynthetic efficiency, CCI, stomatal conductance, protein content and shoot/root nutrient uptakes. Iron and manganese stress conditions were found to enhance the accumulation of secondary metabolites and increase the antioxidant activity such as total polyphenol content (TPC), malondialdehyde (MDA) and superoxide dismutase (SOD). These impacts were more accentuated when Fe and Mn stress were applied simultaneously than when any of the deficiencies was applied alone. More than that, Mn stress alone did not significantly affect the biomass accumulation. The obtained results showed that, in hydroponic conditions, iron and manganese rational fertilization can improve the studied parameters. • A significant negative effect on the photosynthetic efficiency, CCI, stomatal conductance, protein content and shoot/root nutrient uptakes. • Enhancement of the accumulation of secondary metabolites and increase the antioxidant activity such as TPC, MDA and SOD. • These impacts were more accentuated when Fe and Mn stress were applied simultaneously than when any of the deficiencies was applied alone. More than that, Mn stress alone did not significantly affect the biomass accumulation. • In hydroponic conditions, iron and manganese rational fertilization can improve the studied parameters. [ABSTRACT FROM AUTHOR]

Apoptosis resistance is an acquired hallmark of cancer cells and many factors can contribute to the tumor cell apoptosis resistance. In this study, we demonstrated that HECTD3, overexpressed in human esophageal squamous cell carcinoma (ESCC), confers cells resistance to cisplatin-induced apoptosis and promotes cancer cell survival. HECTD3 can bind and ubiquitinate caspase-9, which leads to inhibiting caspase-9 oligomerization and association with Apaf-1, and results in suppressing caspase-9 activation and inhibiting apoptosis. Furthermore, this antiapoptotic function of HECTD3 is dependent on its Thr-157 phosphorylation by ERK. HECTD3, but not T157A mutant, facilitates cell survival in ESCC cells in survival assay in vitro and promotes tumor growth in a xenograft mouse model in vivo. These findings establish a new mechanism of cancer cell resistance to apoptosis and provide a new potential strategy for ESCC treatment. [ABSTRACT FROM AUTHOR]

A three-phase experiment measured the effects of prey quality and fasting on juvenile Chinook salmon ( Oncorhynchus tshawytscha ) performance. The first phase was designed to evaluate the effect of dietary levels of two essential fatty acids, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), on salmon growth. Salmon were reared for 12 weeks on three diets varying in proportions of krill ( Thysanoessa spinifera and Euphausia pacifica ) and northern anchovy ( Engraulis mordax ). Supplements of DHA and EPA were added to the formulated diets to achieve DHA:EPA ratios (0.6, 0.9, and 1.5) representative of naturally occurring prey. Growth rates over 12 weeks were not significantly different among diet treatments, which may be because DHA and EPA were provisioned at or above minimum requirements. Salmon maintained DHA at high levels (> 20% of total fatty acids) across all treatments and sampling periods, whereas EPA reflected dietary concentrations after 12 weeks. Fatty acids were incorporated into salmon muscle at varying rates but on average reflected diet after 1 to 2 months, similar to bulk stable isotopes of carbon and nitrogen. The second phase of the experiment was designed to evaluate fasting effects on salmon size, growth, and lipid storage over 4 weeks. Fed fish were heavier, grew faster, and had significantly more storage lipids than fasted fish. The third phase was designed to evaluate aerobic performance in fasted fish. Critical swim speeds were found to be positively related to salmon body size and storage lipids, but not prior diet quality, evidence that larger salmon with higher energy reserves may be better suited for overwinter survival due to their ability to swim faster than smaller leaner individuals. These results provide mechanistic support for the idea that body condition prior to the first ocean winter is important for juvenile salmon survival. [ABSTRACT FROM AUTHOR]

Background: Zinc has been shown to improve intestinal barrier function against Salmonella enterica serovar Typhimurium (S. typhimurium) infection, but the mechanisms involved in this process remain undefined.Objective: We aimed to explore the roles of G protein-coupled receptor (GPR)39 and protein kinase Cζ (PKCζ) in the regulation by zinc of intestinal barrier function.Methods: A Transwell Caco-2 monolayer was pretreated with 0, 50, or 100 μM Zn and then incubated with S. typhimurium for 0-6 h. Afterward, cells silenced by the small interfering RNA for GPR39 or PKCζ were pretreated with 100 μM Zn and incubated with S. typhimurium for 3 h. Finally, transepithelial electrical resistance (TEER), permeability, tight junction (TJ) proteins, and signaling molecules GPR39 and PKCζ were measured.Results: Compared with controls, S. typhimurium decreased TEER by 62.3-96.2% at 4-6 h (P < 0.001), increased (P < 0.001) permeability at 6 h, and downregulated (P < 0.05) TJ protein zonula occludens (ZO)-1 and occludin by 104-123%, as well as Toll-like receptor 2 and PKCζ by 35.1% and 75.2%, respectively. Compared with S. typhimurium-challenged cells, 50 and 100 μM Zn improved TEER by 26.3-60.9% at 4-6 h (P < 0.001) and decreased (P < 0.001) permeability and bacterial invasion at 6 h. A total of 100 μM Zn increased ZO-1, occludin, GPR39, and PKCζ 0.72- to 1.34-fold (P < 0.05); however, 50 μM Zn did not affect ZO-1 or occludin (P > 0.1). Silencing GPR39 decreased (P < 0.05) zinc-activated PKCζ and blocked (P < 0.05) the promotion of zinc on epithelial integrity. Furthermore, silencing PKCζ counteracted the protective effect of zinc on epithelial integrity but did not inhibit GPR39 (P = 0.138).Conclusion: We demonstrated that zinc upregulates PKCζ by activating GPR39 to enhance the abundance of ZO-1, thereby improving epithelial integrity in S. typhimurium-infected Caco-2 cells. [ABSTRACT FROM AUTHOR]

Cowpea severe mosaic virus (CPSMV) causes significant losses in cowpea ( Vigna unguiculata ) production. In this present study biochemical, physiological, and proteomic analysis were done to identify pathways and defense proteins that are altered during the incompatible interaction between the cowpea genotype BRS-Marataoã and CPSMV. The leaf protein extracts from mock- (MI) and CPSMV-inoculated plantlets (V) were evaluated at 2 and 6 days post-inoculation (DPI). Data support the assumptions that increases in biochemical (high hydrogen peroxide, antioxidant enzymes, and secondary compounds) and physiological responses (high photosynthesis index and chlorophyll content), confirmed by label-free comparative proteomic approach, in which quantitative changes in proteasome proteins, proteins related to photosynthesis, redox homeostasis, regulation factors/RNA processing proteins were observed may be implicated in the resistance of BRS-Marataoã to CPSMV. This pioneering study provides information for the selection of specific pathways and proteins, altered in this incompatible relationship, which could be chosen as targets for detailed studies to advance our understanding of the molecular, physiological, and biochemistry basis of the resistance mechanism of cowpea and design approachs to engineer plants that are more productive. Biological significance This is a pioneering study in which an incompatible relationship between a resistant cowpea and Cowpea severe mosaic virus (CPSMV) was conducted to comparatively evaluate proteomic profiles by Gel-free/label-free methodology and some physiological and biochemical parameters to shed light on how a resistant cowpea cultivar deals with the virus attack. Specific proteins and associated pathways were altered in the cowpea plants challenged with CPSMV and will contribute to our knowledge on the biological process tailored by cowpea in response to CPSMV. [ABSTRACT FROM AUTHOR]

The renin-angiotensin system (RAS) is involved in the development of left ventricular hypertrophy (LVH) by which increases cardiac morbidity and mortality. Activation of mitogen-activated protein kinases (MAPKs) and oxidative stress are important in RAS-mediated cardiac hypertrophy. Sesame oil, a potent antioxidant, attenuates hypertension-dependent LVH. We examined the protective role of sesame oil on RAS-mediated MAPK activation and oxidative stress in rats. We induced LVH using a hypertensive model by subcutaneously injecting deoxycorticosterone acetate (DOCA; 15 mg/ml/kg in mineral oil; twice weekly for 5 weeks) and supplementing with 1% sodium chloride drinking water (DOCA/salt) to uninephrectomized rats. Sesame oil was gavaged (0.5 or 1 ml/kg/day for 7 days) after 4 weeks of DOCA/salt treatment. Cardiac histopathology, RAS parameters, expression of MAPKs, reactive oxygen species and lipid peroxidation were assessed 24 h after the last dose of sesame oil. Sesame oil significantly decreased the size of cardiomyocytes and the levels of cardiac renin, angiotensin-converting enzyme and angiotensin II. In addition, sesame oil down-regulated the expression of angiotensin type 1 receptor, JNK and p38 MAPK and apoptosis signal regulating kinase 1, c-Fos and c-Jun in rats receiving DOCA/salt. Furthermore, the induction of nicotinamide adenine dinucleotide phosphate oxidase, superoxide anion and hydroxyl radical and lipid peroxidation by DOCA/salt were inhibited by sesame oil. Sesame oil modulates cardiac RAS to ameliorate LVH by inhibiting MAPK activation and lowering oxidative stress. [ABSTRACT FROM AUTHOR]

Multiple applications of metal oxide nanoparticles (MONPs) could result in their accumulation in soil, threatening higher terrestrial plants. Several reports have shown the effects of MONPs on plants. In this review, we analyze the most recent reports about the physiological and biochemical responses of plants to stress imposed by MONPs. Findings demonstrate that MONPs may be taken up and accumulated in plant tissues causing adverse or beneficial effects on seed germination, seedling elongation, photosynthesis, antioxidative stress response, agronomic, and yield characteristics. Given the importance of determining the potential risks of MONPs on crops and other terrestrial higher plants, research questions about field long-term conditions, transgenernational phytotoxicity, genotype specific sensitivity, and combined pollution problems should be considered. [ABSTRACT FROM AUTHOR]

Background: Telomere length and telomere shortening are associated with age-related health outcomes. Only a few studies have been able to longitudinally report on factors that are associated with changes in telomere length in an aging population. Objective: We studied the longitudinal relation between telomere length, the change in telomere length, and circulating amino acids. Design: A total of 812 subjects from the Helsinki Birth Cohort Study (born from 1934 to 1944), who underwent 3 clinical visits during a 10-y interval that included measurements of cardiometabolic risk factors, were included in the study. Leukocyte telomere length (LTL) was measured with the use of quantitative real-time polymerase chain reaction. Circulating branched-chain and aromatic amino acids (alanine, glycine, histidine, phenylalanine, leucine, isoleucine, valine, and tyrosine) were assessed with the use of high-throughput nuclear magnetic resonance spectroscopy. Results: The relative ± SD LTL at a mean age of 71 y was 0.79 ± 0.27 in men and 0.89 ± 0.35 in women (P < 0.001). Of the studied amino acids, the strongest inverse association was observed between the phenylalanine concentration that was measured 5 y earlier and the LTL. This finding was significant in men (P = 0.021) and remained significant after adjustment for multiple comparisons, but it was not significant in women (P = 0.39). Longitudinally, the change in LTL over 10 y was inversely associated with the phenylalanine concentration in men (P = 0.007) but not in women (P = 0.58) after adjustment for baseline LTL, age, smoking, and percentage of body fat. Conclusions: The serum phenylalanine concentration is associated with telomere length and, therefore, potentially with the aging process. Because the associations reported are observational, no conclusions can be made regarding causality. Our findings support the hypothesis that cellular pathways that regulate aging are sex specific. [ABSTRACT FROM AUTHOR]

Numerous studies demonstrate that reactive aldehydes are highly toxic and aldehyde dehydrogenase 2 (ALDH2)-mediated detoxification of reactive aldehydes is thought as an endogenous protective mechanism against reactive aldehydes-induced cell injury. This study aims to explore whether lipoic acid, a potential ALDH2 activator, is able to protect gastric mucosa from ethanol-induced injury through a mechanism involving clearance of reactive aldehydes. The rats received 60% of acidified ethanol through intragastric administration and held for 1 h to establish a mucosal injury model. Lipoic acid (10 or 30 mg/kg) or Alda-1 (a positive control, 10 mg/kg) was given 45 min before the ethanol treatment. The gastric tissues were collected for analysis of gastric ulcer index, cellular apoptosis, 4-hydroxy-2-nonenal (4-HNE) and malondialdehyde (MDA) contents, and ALDH2 activity. The results showed that acute administration of ethanol led to an increase in gastric ulcer index, cellular apoptosis, 4-HNE and MDA contents concomitant with a decrease in ALDH2 activity; these phenomena were reversed by lipoic acid or Alda-1. The gastric protection of lipoic acid was attenuated in the presence of ALDH2 inhibitor. Based on these observations, we conclude that lipoic acid exerts the beneficial effects on ethanol-induced injury through a mechanism involving, at least in part, ALDH2 activation. As a dietary supplement or a medicine already in some countries, lipoic acid can be used to treat the ethanol - induced gastric mucosal injury. [ABSTRACT FROM AUTHOR]

Low-intensity ultrasound therapy has been found to be a potential tool in the management of malignant tumors in recent years. However, the molecular mechanism underlying low-intensity ultrasound-induced apoptosis is still not clear. In this study, we investigated the effects of low-intensity ultrasound-induced apoptosis in HEp-2 cells. We found that low-intensity ultrasound significantly induced apoptosis, and the expression level of caveolin-1 (Cav-1) was dramatically increased after ultrasound treatment of HEp-2 cells. After inhibiting the expression level of Cav-1 using siRNA transfection, we found that the cellular apoptosis induced by low-intensity ultrasound was significantly suppressed. In addition, inhibition of Cav-1 expression promoted phosphorylation of signal transducer and activator of transcription 3 (STAT3), suggesting that the STAT3 signaling pathway was involved in low-intensity ultrasound-induced apoptosis via Cav-1 regulation. Our results indicate that Cav-1/STAT3 signaling pathway may mediate low-intensity ultrasound-induced apoptosis, and this technology could potentially be used clinically for the treatment of cancers. [ABSTRACT FROM AUTHOR]

The urea-retaining strategy of elasmobranchs has shaped their biochemistry and physiology; from their metabolic organization to the structure of their membranes. It has also affected their capacity to live in freshwater. Although much new information has been uncovered in the past 30 years, many unanswered questions remain. These include: a) why was urea selected as the major organic osmolyte, b) why is glutamine used as a nitrogen donor, c) why was plasma albumin lost in marine elasmobranchs, d) what membranes are involved in urea retention in the gills, e) how do urea and trimethylamine N-oxide (TMAO) affect membranes, and f) why retain urea in freshwater. Hypotheses are presented for future investigations but some questions may require a time machine to answer. [ABSTRACT FROM AUTHOR]

Objective: To determine the expression and function of the microRNA-29 family (miRNA-29a, miRNA-29b, miRNA-29c) in human leiomyoma and myometrium.Design: Basic science experimental design.Setting: Academic medical center.Patient(s): Women undergoing surgery for symptomatic uterine fibroids.Intervention(s): Overexpression and knockdown of miRNA-29a, miRNA-29b, and miRNA-29c in primary leiomyoma and myometrial cells.Main Outcome Measure(s): [1] Expression of the miRNA-29 family members in vivo in leiomyoma versus myometrium; [2] Major fibrillar collagen (I, II, III) expression in leiomyoma and myometrial cells with manipulation of miRNA-29 species.Result(s): Members of the miRNA-29 family (29a, 29b, 29c) are all down-regulated in leiomyoma versus myometrium in vivo. The expression of the miRNA-29 family can be successfully modulated in primary leiomyoma and myometrial cells. Overexpression of the miRNA-29 family in leiomyoma cells results in down-regulation of the major fibrillar collagens. Down-regulation of the miRNA-29 species in myometrium results in an increase in collagen type III deposition.Conclusion(s): The miRNA-29 family is consistently down-regulated in leiomyoma compared to matched myometrial tissue. This down-regulation contributes to the increased collagen seen in leiomyomas versus myometrium. When miRNA-29 members are overexpressed in leiomyoma cells, protein levels of all of the major fibrillar collagens decrease. The miRNA-29 members are potential therapeutic targets in this highly prevalent condition. [ABSTRACT FROM AUTHOR]

Ethanol at low concentrations (<4%) can serve as a food source for fruit fly Drosophila melanogaster, whereas at higher concentrations it may be toxic. In this work, protective effects of dietary alpha-ketoglutarate (AKG) against ethanol toxicity were studied. Food supplementation with 10-mM AKG alleviated toxic effects of 8% ethanol added to food, and improved fly development. Two-day-old adult flies, reared on diet containing both AKG and ethanol, possessed higher alcohol dehydrogenase (ADH) activity as compared with those reared on control diet or diet with ethanol only. Native gel electrophoresis data suggested that this combination diet might promote post-translational modifications of ADH protein with the formation of a highly active ADH form. The ethanol-containing diet led to significantly higher levels of triacylglycerides stored in adult flies, and this parameter was not altered by AKG supplement. The influence of diet on antioxidant defenses was also assessed. In ethanol-fed flies, catalase activity was higher in males and the levels of low molecular mass thiols were unchanged in both sexes compared to control values. Feeding on a mixture of AKG and ethanol did not affect catalase activity but caused a higher level of low molecular mass thiols compared to ethanol-fed flies. It can be concluded that both a stimulation of some components of antioxidant defense and the increase in ADH activity may be responsible for the protective effects of AKG diet supplementation in combination with ethanol. The results suggest that AKG might be useful as a treatment option to neutralize toxic effects of excessive ethanol intake and to improve the physiological state of D. melanogaster and other animals, potentially including humans. [ABSTRACT FROM AUTHOR]

The oncoprotein MDM2 is an E3 ubiquitin ligase that targets tumor suppressor p53 for ubiquitination and proteasomal degradation, restraining the potent activity of p53 and enabling cell survival and proliferation. Dysregulation of MDM2-p53 axis was frequently observed in human cancers. Originally, it is proposed that MDM2 degradation was mainly achieved by destructive self-ubiquitination. However, recent study suggests that MDM2 may be targeted for degradation by an external E3 ubiquitin ligase(s) under physiological levels. Here, we identified E3 ubiquitin ligase RNF12 as an MDM2-interacting protein through yeast two hybrid methods. We demonstrated that RNF12 targets MDM2 for ubiquitination and proteasomal-dependent degradation, which is independent of MDM2's self-ubiquitination activity. Accordingly, RNF12 elevates p53 protein level by abrogating MDM2-mediated p53 degradation and ubiquitination. Finally, we showed that RNF12 regulates cell growth suppression and DNA damage-induced apoptosis in a p53-dependent manner. Taken together, we establish RNF12 as a novel positive regulator of p53 pathway and an external E3 ubiquitin ligase for MDM2 destruction. These data shed light on the potential roles of RNF12 in MDM2-p53 axis and tumor suppression. [ABSTRACT FROM AUTHOR]

Nectin-4 is a Ca(2+)-independent immunoglobulin-like cell adhesion molecule which has diverse functions in cell-cell adhesion via homophilic and heterophilic interactions. Cell-cell adhesive processes are central to cell polarization, differentiation, proliferation, survival and movement. Here we report that Nectin-4 is substantially overexpressed in gallbladder cancer (GBC), the most common biliary tract malignancy with a high risk of local tumor spread and invasion. Further, Nectin-4 high expression in GBC patients was associated with pathologic T stage and lymph node metastasis status, and the expression level of the downstream target Rac1 and poor prognoses were also correlated with Nectin-4. Ectopic expression of Nectin-4 promoted GBC cell growth, motility and tumor growth in a mouse model. The depletion of Nectin-4 inhibited GBC cell proliferation and migration both in cell culture and in mice. Our data suggest that activation of the PI3K/AKT pathway was involved in the oncogenic function of Nectin-4 to activate Rac1 in GBC. Inhibition of PI3K/AKT with LY294002 and/or Rac1 with NSC23766 impaired Nectin-4-mediated GBC cell proliferation and motility. We hypothesize that Nectin-4 is critical for GBC progression via PI3K/AKT pathway activation of Rac1. Nectin-4 may be a novel prognostic factor and therapeutic target in GBC patients. [ABSTRACT FROM AUTHOR]

Aim To test the hypothesis that changes in enamel component volumes (mineral, organic, and water volumes, and permeability) are graded from outer to inner enamel after a short bleaching procedure. Materials and methods Extracted unerupted human third molars had half of their crowns bleached (single bleaching session, 3 × 15 min), and tooth shade changes in bleached parts were analyzed with a spectrophotometer. Ground sections were prepared, component volumes and permeability were quantified at histological points located at varying distances from the enamel surface ( n = 10 points/location), representing conditions before and after bleaching. Results Tooth shade changes were significant ( p < 0.001; 95% CI = −1/−8; power = 99%), and most of the enamel layer was unaffected after bleaching, except at the outer layers. Multiple analysis of covariances revealed that most of the variance of the change in enamel composition after bleaching was explained by the combination of the set of types of component volume (in decreasing order of relevance: mineral loss, organic gain, water gain, and decrease in permeability) with the set of distances from the enamel surface (graded from the enamel surface inward) (canonical R 2 = 0.97; p < 0.0001; power > 99%). Conclusions Changes in enamel composition after a short bleaching procedure followed a gradient within component volumes (mineral loss > organic gain > water gain > decrease in permeability) and decreased from the enamel surface inward. [ABSTRACT FROM AUTHOR]

Epithelial to mesenchymal transition (EMT) has received considerable attention as a conceptual paradigm for explaining the invasive and metastatic behavior of cells during cancer progression. Our previous study showed that loss of expression of TSSC3 is positively associated with osteosarcoma malignancy and progression. However, whether TSSC3 mediates EMT in osteosarcoma is poorly understood. In the present study, we determined that TSSC3 downregulation induced cell migration and invasion ability and promoted mesenchymal transition of osteosarcoma cells by upregulating mesenchymal markers and inhibiting the epithelial markers. Furthermore, TSSC3 downregulation elicited a signaling cascade that included increased levels of Wnt3a and LRP5, inactivation of GSK-3β, accumulation of nuclear β-catenin and Snail, the augmented binding of β-catenin to TCF-4, and accordingly increased the expression of Wnt target genes (CD44, MMP7). The gene knockdown of these signaling proteins could inhibit TSSC3 downregulation-promoted EMT, migration, and invasion in osteosarcoma. Finally, TSSC3 overexpression obviously inhibited cell migration, invasion, and repressed mesenchymal phenotypes, reducing lung metastasis through GSK-3β activation. Collectively, TSSC3 downregulation promotes the EMT of osteosarcoma cells by regulating EMT markers via a signal transduction pathway that involves Snail, Wnt-β-catenin/TCF, and GSK-3β. [ABSTRACT FROM AUTHOR]

We have previously reported the identification of a rhodanine compound ( 1 ) with well-balanced inhibitory activity against IKKβ and collagen-induced TNFα activated cells. However, we need more optimized compounds because of its instability over plasma and microsome. As part of a program directed toward the optimization of IKKβ inhibitor, we modified a substituent of parent compound to a series of functional groups. Among substituted compounds, fluorine substituent ( 12 ) on the para position of phenyl ring restored the stability toward plasma and microsome while retaining inhibitory potency and selectivity against IKKβ over other kinases. Also, we have demonstrated that compound 12 is an ATP non-competitive inhibitor and safe enough to apply to animal experiment from an acute toxicity test. [ABSTRACT FROM AUTHOR]

Outer membrane protein A (OmpA) plays multiple roles in the physiology and pathogenesis of the zoonotic pathogen enterohemorrhagic Escherichia coli (EHEC). The N-terminus of OmpA forms a transmembrane domain (OmpA™), and the roles of this domain in bacterial pathogenesis have been well studied. However, how its C-terminal domain (OmpAper), which is located at the periplasmic space in the bacterial membrane, contributes to virulence remains unclear. Herein, we report that OmpAper forms a dimer and binds to peptidoglycan in vitro . Furthermore, OmpAper is responsible for bacterial resistance to acidic conditions, high osmotic pressure and high SDS environments. In addition, OmpAper contributes to the adhesion of bacteria to HeLa cells in vitro and ex vivo . These results provide an additional understanding of the role of OmpA in EHEC physiology and pathogenesis. [ABSTRACT FROM AUTHOR]

Objective: To compare the effect of shifting anaerobic to aerobic metabolism on key regulators of oxidative stress, including extracellular superoxide dismutase (SOD3), inducible nitric oxide synthase (iNOS), and its product, nitric oxide (NO), as well as mitochondrial potential (Δψm) and apoptosis in fibroblasts established from normal peritoneum and adhesion tissues.Design: Prospective, experimental study.Setting: University medical center.Patient(s): Fibroblasts established from normal peritoneum and adhesion tissues from the same patients.Intervention(s): Treatment with dichloroacetate (0, 20, 40, and 80 μg/mL, 24 hours).Main Outcome Measure(s): The expression of SOD3, iNOS, and NO levels were measured utilizing real-time reverse transcription-polymerase chain reaction and Greiss assay. The Δψm was evaluated by the JC-1 Mitochondrial Membrane Potential Assay. Apoptosis was determined by caspase-3 activity and TUNEL assays. Data were analyzed using SPSS 19.0. Mixed model repeated-measures analysis of variance was used with a Bonferroni correction. Significant interactions were analyzed with independent sample t tests.Result(s): Dichloroacetate increased apoptosis, SOD3 messenger RNA, iNOS messenger RNA, and NO levels in fibroblasts from peritoneum and adhesions. There was enhanced Δψm adhesion as compared with normal peritoneal fibroblasts. Creating oxidative stress by exposure by hypoxia markedly increased Δψm in fibroblasts from normal peritoneum to levels observed in adhesions; dichloroacetate protected against the effects of hypoxia.Conclusion(s): Anaerobic metabolism and oxidative stress are associated with the development of the adhesion phenotype, which manifests decreased apoptosis. Dichloroacetate induces adhesion fibroblasts to undergo apoptosis via modulation of redox homeostasis. These findings may provide targets for therapeutic treatment for reduction of profibrotic disorders, including postoperative adhesions. [ABSTRACT FROM AUTHOR]