Concentration-dependent Way Research Articles

Zinc oxide nanoparticles enhance expression of maspin in human breast cancer cells.

Toxic and apoptotic impacts of zinc oxide nanoparticle (ZNP) on different cancer cells have been reported. Maspin (a mammary serine protease inhibitor) as a tumor suppressor gene can inhibit tumor growth and metastasis. The expression of maspin is modulated by p53, Bcl-2 family genes, and estrogen receptor α (ER-α). This study aimed to assess the ZNP effects on maspin expression in MCF-7 cells (a breast cancer cell). Experimental groups (ZNP5, ZNP10, and ZNP20) received 5, 10, and 20μM/mL ZNP for 48h, respectively. 17-β-estradiol (E2) was used to evaluate the role of ER-α in the anticancer impact of ZNP. Cell viability, Annexin V, migration assay, gene expression, and western blotting methods were applied to evaluate ZNP effects on the MCF-7 cells. ZNP at the concentrations of 10 and 20μM/mL could significantly decrease the viability and migration rate, and significantly increase apoptosis percentage in the MCF-7 cells. ZNP significantly enhanced mRNA expression and protein level of maspin in MCF-7 cells in a concentration-dependent way. ZNP concentration-dependently elevated mRNA expression and protein level of p53 and Bax while reduced the expression of Bcl-2 and ER-α. E2 promoted cancer cell growth by enhancing survival and migration rates. E2 treatment reduced mRNA expression and protein level of maspin and p53, and elevated Bcl-2 expression. ZNP considerably changed these events induced by E2 in the MCF-7 cells. It is concluded that the maspin overexpression is one of the toxic mechanisms of the ZNP on the ER-α-positive breast cancer cells, and can suppress the migration of these cells.

Antioxidant, Anti-Inflammatory and Antiproliferative Effects of the Vitis vinifera L. var. Fetească Neagră and Pinot Noir Pomace Extracts.

The pathophysiology of inflammation and oxidative stress generated during different types of cancers and anticancer treatments is well documented. Traditionally, grape pomace is used for animal feed, organic fertilizers, ethanol production or is disposed as waste. Because grape pomace is a rich source of antioxidant compounds, the purpose of the study was to evaluate the antioxidant, anti-inflammatory, and antiproliferative effects of fresh and fermented grape pomace extracts of two Vitis vinifera L. varieties Fetească neagră and Pinot noir cultivated in Romania. Firstly, grape pomace phytochemical analysis and in vitro antioxidant tests were performed. Secondly, the effect of a seven-day pretreatment with grape pomace extracts on the turpentine oil-induced inflammation in rats was assessed by measuring total oxidative status, total antioxidant response, oxidative stress index, malondialdehyde, total thiols, nitric oxide and 3-nitrotyrosine. Thirdly, the antiproliferative properties were evaluated on human lung carcinoma (A549), human breast adenocarcinoma (MDA-MB-231), murine melanoma (B164A5), and keratinocyte (HaCat) cell lines. Fetească neagră and Pinot noir grape pomace extracts have a rich content of polyphenols and in vitro antioxidant effect. Fermented samples had higher polyphenol content, but fresh samples had better antioxidant activity. Pretreatment with grape pomace extracts reduced inflammation-induced oxidative stress in a concentration-dependent way, fresh samples being more efficient. The malignant cells’ proliferation was inhibited by all grape pomace extracts, fermented Fetească neagră extracts having the strongest effect. Conclusion: fresh and fermented pomace extracts of Vitis vinifera L. varieties Fetească neagră and Pinot noir cultivated in a Romanian wine region have antioxidant, anti-inflammatory and antiproliferative effects.

Physiological and morphological response of Aphanizomenon flos-aquae to watermelon (Citrullus lanatus) peel aqueous extract

Natural algaecides are more likely to be specific and biodegradable, and may offer an environmentally friendly method for control of cyanobacterial blooms. We explored, for the first time, the potential for watermelon peel aqueous extract (WMPAE) to control the growth of the harmful blue-green alga Aphanizomenon flos-aquae. The growth inhibition and several physiological parameters of A. flos-aquae, in response to WMPAE, were analyzed. Results showed that WMPAE significantly inhibited the growth of A. flos-aquae in a concentration-dependent way. The highest inhibition reached 94 % after 3 days’ treatment with 6 g L−1 of WMPAE and a significant effect was obtained with lower doses and shorter times as well. The cell viability decreased quickly, cell shape changed, and intracellular structural damage occurred. At the same time, the antioxidant enzymes (superoxide dismutase SOD, catalase CAT and peroxidase POD) and malondialdehyde (MDA) levels all increased significantly, indicating that WMPAE between 2–6 g L−1 induced severe oxidative stress and damage to A. flos-aquae. Moreover, production of the four pigments chlorophyll a (Chl a), carotenoids, phycocyanin (PC), and allophycocyanin (APC) were all stimulated, though photosynthesis of A. flos-aquae was clearly inhibited. The maximum quantum yield of photosystem II (Fv/Fm) and the effective quantum yield of photosystem II ( Fv'/Fm') declined sharply, suggesting the decreased photosystem capacity of A. flos-aquae to convert light energy into chemical energy. In addition, non-photochemical quenching (NPQ) of A. flos-aquae increased after a very short time exposure to WMPAE, and decreased significantly with prolonged exposure time, which indicated the failure of photo protection mechanisms. These results suggest that the loss of cell viability, and increases in oxidative stress, and damage to intracellular structure and photosynthetic systems might be the mechanisms for the inhibitory effects. Our results suggested that WMPAE could be a novel and effective approach for controlling the growth of A. flos-aquae in aquatic environments.

Multiplexed Bioanalysis Based on Light-sensitive Electrodes

Introduction Quantum dots (QDs) have become attractive tools in bioanalysis because of their advantageous optical and electrochemical properties. This relates to applications exploiting fluorescence, electrochemiluminescence or photocurrent generation [1]. For the latter case QDs are immobilized on conductive electrodes. By light excitation charge carriers are formed inside the nanoparticles which can be used for photocurrent generation. Signal chains can be constructed when appropriate donor or acceptor components are present in solution. The special feature of such electrodes is that the response is triggered by illumination of the sensing electrode [1]. In this study, a light-addressable photoelectrochemical sensor was built by coupling QDs with oxidases to detect different enzyme substrates. Results and Conclusions At first CdSe/ZnS QDs have been immobilised via a dithiol compound on gold electrodes. Two different enzymes, glucose oxidase (GOD) and sarcosine oxidase (SOD) have been deposited on this QDs-electrode at spatially well resolved places. Here small spots have been achieved by a dropping method. A laser beam is moved over the spots to detect photocurrents. Since oxygen can act as electron acceptor at the illuminated QDs the photocurrent is correlated to the oxygen concentration in solution. By action of the enzymes in the presence of the corresponding substrate a competitive situation for oxygen is created in front of the QD layer and thus, the photocurrent decreases [2, 3]. Since this happens in a concentration dependent way, analysis of the enzyme substrate is feasible.Furthermore, due to the spatially separated immobilisation of the two enzymes, an independent read-out of the two enzyme reactions is possible. The results indicate good lateral resolution and selectivity of the multi-detection system without any crosstalk between the two sensing regions and also without any response at regions, which have not been modified with an enzyme [4].After that, the possibility of an imaging of the enzyme spot by photocurrent measurement has been studied. When the laser is moved over the spot of GOD with a constant speed, the size of the spot can be determined from the photocurrent - time curve (with a defined moving speed). The photocurrent correlates to different local enzyme activities. Consequently, an imaging of the enzyme activity distribution on the surface has also been achieved by this method.

Roflumilast, a Phosphodiesterases-4 (PDE4) Inhibitor, Alleviates Sepsis‑induced Acute Kidney Injury.

BackgroundSepsis causes acute kidney injury (AKI) in critically ill patients. Roflumilast, a phosphodiesterases-4 (PDE4) inhibitor, has been shown to be therapeutically effective in sepsis-induced organ injury. However, the function of roflumilast in sepsis-induced AKI is not clearly understood. The present study aimed to explore the protective effect of roflumilast on sepsis-induced AKI in mice.Material/MethodsA sepsis model was established by cecal ligation and puncture surgery. Roflumilast (1 mg/kg and 3 mg/kg) was used once daily for 7 consecutive days for treatment. Kidney tissues were pathologically examined by hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS) staining. The levels of kidney injury markers including blood urea nitrogen (BUN), creatinine (Cre), kidney injury molecule-1 (KIM-1), and neutrophil gelatinase-associated lipocalin (NGAL), and inflammatory cytokines including interleukin (IL)-6, tumor necrosis factor (TNF)-α, and IL-1β were detected by their corresponding test kits. The protein expression was measured using western blot and cell apoptosis of kidney tissue was determined by TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assay.ResultsRoflumilast was demonstrated to alleviate sepsis-induced AKI by reducing histopathological changes and decreasing the levels of kidney injury markers in a concentration-dependent way. The production of TNF-α, IL-6, and IL-1β was significantly suppressed by roflumilast. Besides, roflumilast inhibited the activation of NLRP3 (nucleotide-binding domain (NOD)-like receptor protein 3) and NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells). Additionally, roflumilast inhibited cell apoptosis and changes in expression of apoptosis related proteins induced by sepsis. Finally, high concentration of roflumilast (3 mg/kg) did not have an adverse effect on liver, heart, lung, or spleen.ConclusionsOur study indicated that roflumilast could ameliorate AKI induced by sepsis through restraining inflammatory response and apoptosis of the kidney, providing a molecular basis for a novel medical treatment of septic AKI.

Antispasmodic effect of hydroalcoholic and flavonoids extracts ofDracocephalum kotschyion rabbit bladder

Introduction:Dracocephalum kotschyiextract has antispasmodic activities on smooth muscle including ileum, uterus and trachea. The objective of this research was to investigate antispasmodic activity of hydroalcoholic and flavonoids extracts ofD. kotschyion rabbit bladder contractions.Methods:Rabbits were euthanized by carbon dioxide asphyxiation and the whole bladder was dissected out and immersed in the Tyrode’s solution. Longitudinal bladder strips were mounted vertically in an organ bath at 37°C and gassed continuously with O2. Bladder strips were contracted with acetylcholine (ACh), KCl, or electrical field stimulation (EFS). Isotonic tension of the tissue was recorded before and after addition of hydroalcoholic or flavonoids rich extracts ofD. kotschyi. Nifedipine and propantheline were used as standard drugs.Results:Standard drug propantheline, prevented bladder phasic contraction induced by ACh (1µM) without affecting KCl response. On the other hand, cumulative addition of nifedipine attenuated the tonic contractions induced by KCl (20mM) on bladder smooth muscle. Hydroalcoholic and flavonoids extracts ofD. kotschyiat concentration ranges of 10-320 µg/ mL in a concentration dependent way inhibited bladder tonic contraction induced by KCl (n=6). Both extracts also in a concentration-dependent manner relaxed EFS and ACh-induced contractions (range, 20–1280 µg/mL) of bladder smooth musclein vitro. Complete inhibition was achieved with the highest used concentrations of the extracts. The inhibitory effect of the extract was reversible following washing the tissues with fresh Tyrode’s solution.Conclusion:This study clearly demonstrated thatD. kotschyiextracts were able to prevent contractions induced by ACh, KCl or EFS in isolated rabbit bladder. This means that people consuming this medicinal plant may face urinary retention which could be a problem for patients with prostate hypertrophy. On the other hand, this plant might be useful in patients with urinary incontinence. However, its usefulness must be assessed in the controlled clinical trials.

Volatile anaesthetics inhibit the thermosensitive nociceptor ion channel transient receptor potential melastatin 3 (TRPM3)

BackgroundVolatile anaesthetics (VAs) are the most widely used compounds to induce reversible loss of consciousness and maintain general anaesthesia during surgical interventions. Although the mechanism of their action is not yet fully understood, it is generally believed, that VAs depress central nervous system functions mainly through modulation of ion channels in the neuronal membrane, including 2-pore-domain K+ channels, GABA and NMDA receptors. Recent research also reported their action on nociceptive and thermosensitive TRP channels expressed in the peripheral nervous system, including TRPV1, TRPA1, and TRPM8. Here, we investigated the effect of VAs on TRPM3, a less characterized member of the thermosensitive TRP channels playing a central role in noxious heat sensation. MethodsWe investigated the effect of VAs on the activity of recombinant and native TRPM3, by monitoring changes in the intracellular Ca2+ concentration and measuring TRPM3-mediated transmembrane currents. ResultsAll the investigated VAs (chloroform, halothane, isoflurane, sevoflurane) inhibited both the agonist-induced (pregnenolone sulfate, CIM0216) and heat-activated Ca2+ signals and transmembrane currents in a concentration dependent way in HEK293T cells overexpressing recombinant TRPM3. Among the tested VAs, halothane was the most potent blocker (IC50 = 0.52 ± 0.05 mM). We also investigated the effect of VAs on native TRPM3 channels expressed in sensory neurons of the dorsal root ganglia. While VAs activated certain sensory neurons independently of TRPM3, they strongly and reversibly inhibited the agonist-induced TRPM3 activity. ConclusionsThese data provide a better insight into the molecular mechanism beyond the analgesic effect of VAs and propose novel strategies to attenuate TRPM3 dependent nociception.

Propofol Inhibits Cell Proliferation, Migration, and Invasion via mir-410-3p/Transforming Growth Factor-β Receptor Type 2 (TGFBR2) Axis in Glioma.

BackgroundPropofol is a common intravenous anesthetic used to induce and maintain anesthesia. Numerous studies have reported that propofol plays an anti-tumor role in diverse human cancers, including glioma. In this research, we explored the roles of propofol and its related molecular mechanisms in glioma.Material/MethodsU251 and A172 cells were exposed to different doses of propofol for 24 h. Cell proliferation, migration, and invasion in glioma were evaluated using MTT assay and Transwell assay, respectively. The levels of microRNA-410-3p (miR-410-3p) and transforming growth factor-β receptor type 2 (TGFBR2) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) assay and Western blot assay, respectively. The association between miR-410-3p and TGFBR2 was predicted by TargetScan and confirmed by dual-luciferase reporter assay.ResultsPropofol inhibited the proliferation, migration, and invasion of glioma cells in a concentration-dependent way. miR-410-3p was induced and TGFBR2 was inhibited by different concentrations of propofol treatment. Moreover, TGFBR2 was confirmed to be a target gene of miR-410-3p and TGFBR2 was inversely modulated by miR-410-3p in glioma cells. Depletion of miR-410-3p reversed the inhibition of propofol treatment on U251 and A172 cell growth and metastasis, but the effects were further abolished by knocking down the expression of TGFBR2.ConclusionsPropofol can suppress cell growth and metastasis by regulating the miR-410-3p/TGFBR2 axis in glioma.

Searching for a common origin of heat-transfer effects in bio- and chemosensors: A study on thiols as a model system

The heat-transfer method HTM is a bioanalytical technique in which a temperature gradient is established between the backside of a functionalized chip and the supernatant liquid. By combining the measured temperature difference with the power used to generate this gradient, one obtains the thermal resistance Rth. This parameter responds sensitively and in a concentration-dependent way to the binding of bioparticles to receptors as well as to phase transitions in coatings on the chip. The size of particles that can be detected with HTM spans from low-molecular weight molecules over proteins and DNA fragments up to cells with diameters at the micron scale. In this work, we explore the question whether and why small ligands adsorption can result still in quantifiable Rth changes and whether there is a common origin of the generally observed Rth increase upon binding a wide variety of cells and biomolecules. The data obtained on thiols with different capping groups suggest that the correspondence of molecular vibration frequencies of the ligands and the liquid is decisive for an efficient or impeded heat transfer and hence for the macroscopically determined Rth parameter.

Eupatilin Alleviates Lipopolysaccharide-Induced Acute Lung Injury by Inhibiting Inflammation and Oxidative Stress.

BackgroundEupatilin, an active flavone separated from Artemisia species, has various biological activity such as anti-inflammatory activity. The aim of the present study was to find out the influence of eupatilin on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats.Material/MethodsThe administration of LPS was used to induce ALI; eupatilin was given 1 hour before the LPS administration. Lung structural damage of rats was analyzed by hematoxylin and eosin staining and the wet/dry lung ratio. The related inflammatory factors and lung injury markers were examined by enzyme-linked immunosorbent assay. The oxidative stress factors were analyzed by corresponding kits. The expression of peroxisome proliferator-activated receptor-α (PPAR-α) was assayed by western blot and immunohistochemical staining.ResultsThe results showed that eupatilin alleviated LPS-induced structural damage and decreased the wet/dry lung ratio concentration-dependently. Eupatilin decreased the level of surfactant protein (SP)-A, SP-D, and inflammatory factors such as interleukin (IL)-6, tumor necrosis factor (TNF)-α, and monocyte chemo-attractant protein (MCP)-1. LPS trigged nitric oxide (NO) generation, improved the production of malondialdehyde (MDA) and lactate dehydrogenase (LDH) and decreased the activity of superoxide dismutase (SOD), which were reversed when rats treated with eupatilin in a concentration-dependent way. Besides, the expression of PPAR-α was increased under the treatment of eupatilin.ConclusionsCollectively, eupatilin alleviated LPS-induced ALI through inhibiting inflammation and oxidative stress in a concentration-dependent way, which was likely to be closely related with the activation of PPAR-α.

Eco Friendly Fabrication of Gold Nanoclusters and Their Induction of Cardiomyocyte Apoptosis After Intratracheal Instillation in Rats

Cardiac diseases instigated by the exposure of nanoparticles have gained tremendous attention of research scientists in recent days. After entering the blood stream, nanoparticles have the ability to affect the cardiomyocytes, resulting in the cardiac failure. In this work, the toxicity of biosynthesized gold nanoparticles (AuNPs) on cardiomyocytes of rats was studies. AuNPs were prepared by the reduction of aqueous HAuCl4 using C. viscosa leaf extract. An optical absorption band for AuNPs was found at 540 nm using UV–visible spectrophotometer. Transmission electron microscopy (TEM) results showed the formation of gold nanocluster with size existing in the range of 1 to 1.5 nm. Also, Fourier transform infrared (FTIR) spectroscopic results confirmed the capping of AuNPs with C. viscosa leaf extract polyphenols. Further, toxicity studies revealed that the levels of gold in the serum and heart, and myocardial enzymes in treated animals were suggestively improved in a concentration-dependent way. Furthermore, treatment with AuNPs resulted the prominent ultrastructural and histological changes in the hearts of treated rats. Additionally, a substantial apoptosis was found in the treated animals. The obtained results indicated that AuNPs has the possibility to enter the circulatory system via lungs, and are circulated to heart resulting the cardiovascular injury. AuNPs-persuaded apoptosis through mitochondrial pathway may play a key role in inducing cardiac failure.

Revealing Protein Aggregates under Thapsigargin-Induced ER Stress Using an ER-Targeted Thioflavin.

Endoplasmic reticulum-thioflavin T (ER-ThT), a thioflavin T-based fluorescent chemosensor, was developed to detect protein aggregates in the endoplasmic reticulum (ER) and was applied to live cells under various forms of ER stress. Upon dithiothreitol (DTT)-induced reductive denaturation of lysozyme and albumin, the intensity was increased in a protein concentration-dependent way, following a nonfluorescent lag phase. ER-ThT detects protein aggregates rather than unfolded proteins in solution, and the protein aggregation can be visualized in the presence of lipid membranes or native proteins. Within live HeLa cells, ER-ThT is localized in the ER and its fluorescence was dramatically increased upon ER stress induction by DTT, Thapsigargin, or Brefeldin A. Moreover, in the presence of ER stress modulators (tauroursodeoxycholic acid, trimethylamine N-oxide, or 4-phenylbutyric acid), also known as chemical chaperones, the fluorescence under Thapsigargin treatment was suppressed to the level of the control group. Thus, ER-ThT is capable of detecting the accumulation of protein aggregates under ER stress in living cells and acts as an in vitro screening tool for ER stress modulators, putative prodrugs against ER-related proteopathy. Overall, the results strongly suggest that protein aggregation is intricately involved in the activation of the unfolded protein response following ER stress.

Achyranthes bidentata polypeptide k suppresses neuroinflammation in BV2 microglia through Nrf2-dependent mechanism.

Activated microglia play a critical role in regulating neuroinflammatory responses in central nervous system. Previous studies have shown that Achyranthes bidentata polypeptide k's (ABPPk's) neuroprotective effects are partly due to its anti-inflammatory effect, but the mechanism remains unknown. This study is aimed to investigate the anti-inflammatory effect of ABPPk on lipopolysaccharide (LPS)-activated neuroinflammation in BV2 microglia. We pretreated BV2 microglia with different concentrations of ABPPk (0.04-5 µg/mL) for 30 minutes, and then stimulated microglia with LPS for 24 hours. Pro-inflammatory mediators including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), nitric oxide (NO) and prostaglandin E2 (PGE2) production were measured by enzyme-linked immunosorbent assay (ELISA) kits. Inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), phosphorylated nuclear factor kappa B (NF-κB), heme oxygenase-1 (HO-1) and nuclear factor erythroid 2-related factor 2 (Nrf2) expression levels were detected by western blot. Glutathione (GSH) level was measured by GSH-Glo™ Glutathione assay. Immunofluorescent staining was used to detect the nuclear translocation of NF-κB and Nrf2. BV2 microglia transfected with Nrf2 siRNA were used to investigate the effect of Nrf2 on the anti-inflammatory activity of ABPPk. ABPPk (0.2-5 µg/mL) reduced the iNOS mediated NO and COX-2 mediated PGE2 production significantly in LPS-activated BV2 microglia. ABPPk (1 and 5 µg/mL) also suppressed the production of TNF-α and IL-6 significantly. NF-κB is phosphorylated and translocated into nuclear in LPS-activated BV2 microglia, but ABPPk is shown to inhibit the phosphorylation and translocation of NF-κB in a concentration-dependent way. ABPPk increased the protein expression levels of HO-1 and Nrf2, as well as the GSH content in BV2 microglia. Immunofluorescent staining showed that ABPPk also promoted nuclear translocation of Nrf2. After knocking down Nrf2 in BV2 cells with siRNA interference, ABPPk's inhibitory effect on pro-inflammatory mediators also disappeared. The present study suggests that ABPPk inhibits neuroinflammation in BV2 microglia through Nrf2-dependent mechanism. This provides some strong evidence for the potential of this neuroprotective natural compound to treat neurodegenerative diseases such as ischemic stroke and Parkinson's disease.

Giving combined medium-chain fatty acids and glucose protects against cancer-associated skeletal muscle atrophy.

Skeletal muscle volume is associated with prognosis of cancer patients. Maintenance of skeletal muscle is an essential concern in cancer treatment. In nutritional intervention, it is important to focus on differences in metabolism between tumor and skeletal muscle. We examined the influence of oral intake of glucose (0%, 10%, 50%) and 2% medium‐chain fatty acid (lauric acid, LAA, C12:0) on tumor growth and skeletal muscle atrophy in mouse peritoneal metastasis models using CT26 mouse colon cancer cells and HT29 human colon cancer cells. After 2 weeks of experimental breeding, skeletal muscle and tumor were removed and analyzed. Glucose intake contributed to prevention of skeletal muscle atrophy in a sugar concentration‐dependent way and also promoted tumor growth. LAA ingestion elevated the level of skeletal muscle protein and suppressed tumor growth by inducing tumor‐selective oxidative stress production. When a combination of glucose and LAA was ingested, skeletal muscle mass increased and tumor growth was suppressed. Our results confirmed that although glucose is an important nutrient for the prevention of skeletal muscle atrophy, it may also foster tumor growth. However, the ingestion of LAA inhibited tumor growth, and its combination with glucose promoted skeletal muscle integrity and function, without stimulating tumor growth. These findings suggest novel strategies for the prevention of skeletal muscle atrophy.

The allelochemical tannic acid affects the locomotion and feeding behaviour of the pond snail, Lymnaea stagnalis, by inhibiting peripheral pathways

(1) The effect of tannic acid (TA), a dominant component of plant allelochemicals, was investigated on the locomotion and feeding of the pond snail, Lymnaea stagnalis. The effect of TA on the neuronal background underlying feeding activity was also analysed. (2) TA affected the spontaneous locomotion and of juvenile snails in a concentration-dependent way. Low (10 μM) TA concentration resulted in an increased (sliding or swimming) activity compared to the control; meanwhile, high (100 μM) TA concentration inhibited the locomotion of the animals. (3) Low (10 μM) TA concentration increased the frequency of sucrose-evoked feeding of intact animals, whereas high (100 μM) TA concentration resulted in significantly longer feeding latency and decreased feeding rate. The feeding changes proved to be partially irreversible, since after 48 h maintained in clear water, the animals tested in 100 μM TA previously still showed lower feeding rate in sucrose. (4) Electrophysiological experiments on semi-intact preparations showed that application of 100 μM TA to the lip area inhibited the fictive feeding pattern of central neurons, the cellular response to sucrose. (5) On isolated CNS preparation, 100 μM TA applied in the bathing solution, however, failed to inhibit the activation of the central feeding (CPG) interneurons following application of extracellular dopamine. Our results suggest that TA affects both afferent and efferent peripheral functions in Lymnaea. TA reduces feeding activity by primarily blocking feeding sensory pathways, and its negative effect on locomotion may imply sensory pathways and/or ciliary activity.

Microwave-assisted extraction of total saponins from Physalis alkekengi L. var. franchetii (Mast.) Makino and their in vitro anti-inflammatory activity

In this study, the optimization of microwave-assisted extraction of total saponins from P. alkekengi (TSP) and in vitro anti-inflammatory activity were discussed. According to the results in Box–Behnken design, the nonlinear regression model obtained from the Design-Expert software was a reasonable fit for the experiment data (R2 = 0.9994). Results showed that the optimal extraction conditions with a yield of 6.41 ± 0.25 mg/g for crude TSP extraction were aqueous ethanol (80%) to raw material ratios of 32:1 (mL:g), extraction power of 300 W, and extraction time of 29 min. The content of the total saponins in the purified extract was 91.32 ± 1.57%. Bioassay indicated that TSP could significantly inhibited the release of nitric oxide (NO), interleukin-6 (IL-6), and interleukin-1β (IL-1β) induced by lipopolysaccharide (LPS) in a dose-dependent manner, and down-regulated the expressions of related genes. In addition, TSP treatment could also suppress the elevated cyclooxygenase-2 (COX-2) and tumor necrosis factor-α (TNF-α) gene expression in LPS-induced RAW 264.7. Moreover, the secretions of monocyte chemoattractant protein-1 (MCP-1) in stimulated macrophages were also restrained in a concentration-dependent way. All these results indicated that TSP exhibited anti-inflammatory effects by suppressing the transcription of some inflammatory cytokine genes, suggesting that the anti-inflammatory properties of TSP may be helpful to mitigate inflammatory diseases as well as some other related diseases. The overall results indicated that TSP had potential as anti-inflammatory lead compounds for follow-up research.

Intestinal cytotoxicity induced by Escherichia coli is fully prevented by red wine polyphenol extract: Mechanistic insights in epithelial cells

Gastrointestinal infections are considered a serious public health problem in view of their high incidence and the increasing antibiotic resistance of the microorganisms involved in their pathogenesis, namely Escherichia coli. Consequently, finding new ways to prevent and/or threat these infections is urgent.In this study we investigated whether a well-characterised polyphenolic red wine extract is able to inhibit the cytotoxic effects induced by a clinical pathogenic Escherichia coli strain (E. coli 270) against HT-29 colon epithelial cells. Firstly, we provide evidences showing that the E. coli strain triggered the death of the intestinal epithelial cells through the production and release of a toxin. Then we support that, in a concentration dependent way, RWE through both, a direct interaction with bacterial exotoxin and the epithelial cells, prevented the action of the toxin on the cells, significantly reducing cell death.This intends to constitute a position paper as detailed mechanisms for the inhibition of E. coli-induced toxicity by polyphenols are yet to be completely unraveled. However, considering that the amount of red wine polyphenols used can be easily achieved in a normal diet, this study suggests that RWE may provide a readily available dietary product with potential benefit for the prevention and/or treatment of intestinal infections induced by intestinal pathogenic bacteria and may open new therapeutic avenues for the development of potential natural antimicrobial agents.

Physiological and transcriptomic analyses reveal the molecular networks of responses induced by exogenous trehalose in plant.

It is well known that exogenous trehalose can improve resistances of plants to some abiotic and biotic stresses. Nonetheless, information respecting the molecular responses of tobacco leaves to Tre treatment is limited. Here we show that exogenous Tre can rapidly reduce stomatal aperture, up-regulate NADPH oxidase genes and increase O2•-andH2O2 on tobacco leaves at 2 h after treatment. We further demonstrated that imidazole and DPI, inhibitors of NADPH oxidase, can promote recovery of stomatal aperture of tobacco leaves upon trehalose treatment. Exogenous trehalose increased tobacco leaf resistance to tobacco mosaic disease significantly in a concentration-dependent way. To elucidate the molecular mechanisms in response to exogenous trehalose, the transcriptomic responses of tobacco leaves with 10 (low concentration) or 50 (high concentration) mM of trehalose treatment at 2 or 24h were investigated through RNA-seq approach. In total, 1288 differentially expressed genes (DEGs) were found with different conditions of trehalose treatments relative to control. Among them, 1075 (83.5%) were triggered by low concentration of trehalose (10mM), indicating that low concentration of Tre is a better elicitor. Functional annotations with KEGG pathway analysis revealed that the DEGs are involved in metabolic pathway, biosynthesis of secondary metabolites, plant hormone signal transduction, plant-pathogen interaction, protein processing in ER, flavonoid synthesis and circadian rhythm and so on. The protein-protein interaction networks generated from the core DEGs regulated by all conditions strikingly revealed that eight proteins, including ClpB1, HSP70, DnaJB1-like protein, universal stress protein (USP) A-like protein, two FTSH6 proteins, GolS1-like protein and chloroplastics HSP, play a core role in responses to exogenous trehalose in tobacco leaves. Our data suggest that trehalose triggers a signal transduction pathway which involves calcium and ROS-mediated signalings. These core components could lead to partial resistance or tolerance to abiotic and biotic stresses. Moreover, 19 DEGs were chosen for analysis of quantitative real-time polymerase chain reaction (qRT-PCR). The qRT-PCR for the 19 candidate genes coincided with the DEGs identified via the RNA-seq analysis, sustaining the reliability of our RNA-seq data.

Di-(2-ethylhexyl) phthalate induced the growth inhibition and oxidative damage in the microalga Chlorella vulgaris

di-2-ethylhexyl phthalate (DEHP) poses a great threat to aquatic ecosystems, with known hazards to aquatic species. The ecotoxicological mechanism of DEHP in ocean microalgae is not yet known. In this study, we investigated growth inhibition, oxidative damage and antioxidant enzyme activities in Chlorella vulgaris under DEHP treatment. After 5-d exposure to DEHP ranging from 2 to 10 mg/L, the growth of C. vulgaris was significantly inhibited. The 96-h median effective concentration values (96h-EC50) of DEHP for C. vulgaris were 6.02 mg/L. This growth inhibition was associated with DEHP-induced oxidative stress, as evidenced by elevated levels of intracellular malondialdehyde (MDA) and hydrogen peroxide. The results showed that DEHP reduced superoxide dismutase and glutathione peroxidise activities, increased hydrogen peroxide level and MDA content in a concentration-dependent way. These results indicated that DEHP could have biochemical and physiological toxic effects in C. vulgaris. These findings will help to understand the toxicity mechanisms of DEHP and the environmental risk assessment of primary producers of aquatic ecosystems

How to avoid severe coronary vasoconstriction in potassium induced cardioplegia

Objectives. The aim of this study was to investigate how high K+ concentrations can be safely used in cardioplegic solutions without causing severe coronary artery vasocontraction.Design. Twenty-four 50 kg pigs were used. The distal part of the left anterior descending coronary artery was cut into ring segments and transferred into organ baths with Krebs solution bubbled with 95% O2 and 5% CO2. K+ concentrations between 16 and 127 mM were used to induce vasocontractions at 37, 22, 15, and 8 °C. Mg2+ (0–20 mM) were used to attenuate K+ induced vasocontractions.Results. K+-Krebs solution 127 mM at 37 °C induced a strong, sustained vasocontraction defined as 100%. The contractions induced by 16, 23, 30 and 127 mM K+ were: 7.7, 38, 72 and 100% at 37 °C; 1.7, 7.4, 21 and 65% at 22 °C; 1, 6.6, 15 and 33% at 15 °C; 0.6, 2.1, 6 and 14% at 8 °C, respectively. Mg2+ reduced the K+-induced contraction at 37 °C in a concentration-dependent way and Mg2+ at 8 mM practically eliminated the risk for severe vasocontraction.Conclusions. Hypothermia (8 °C) abolishes coronary contraction induced by K+-cardioplegic solutions. In normothermic cardioplegia 8 mM Mg2+ prevents vasoconstriction.