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Summary: The Janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) signaling pathway plays central roles in cancer cell growth and survival. Drug repurposing strategies have provided a valuable approach for developing antitumor drugs. Zelnorm (tegaserod maleate) was originally designed as an agonist of 5-hydroxytryptamine 4 receptor (5-HT4R) and approved by the FDA for treating irritable bowel syndrome with constipation (IBS-C). Through the use of a high-throughput drug screening system, Zelnorm was identified as a JAK/STAT3 signaling inhibitor. Moreover, the inhibition of STAT3 phosphorylation by Zelnorm was independent of its original target 5-HT4R. Zelnorm could cause G1 cell cycle arrest, induce cell apoptosis and inhibit the growth of a variety of cancer cells. The present study identifies Zelnorm as a novel JAK/STAT3 signaling inhibitor and reveals a new clinical application of Zelnorm upon market reintroduction. [ABSTRACT FROM AUTHOR]

α-L-Fucosidase (AFU) is a promising therapeutic target for the treatment of inflammation, cancer, cystic fibrosis, and fucosidosis. Some of the existing analytical methods for the assessment of AFU activity are lacking in sensitivity and selectivity, since most of them are based on spectrofluorimetric methods. More recently, mass spectrometry (MS) has evolved as a key technology for enzyme assays and inhibitor screening as it enables accurate monitoring of the conversion of substrate to product in enzymatic reactions. In this study, UHPLC-MS has been utilized to develop a simple, sensitive, and accurate assay for enzyme kinetics and inhibition studies of AFU3, a member of the AFU family. A reported method for analyzing saccharide involving a porous graphitic carbon column, combined with reduction by NaBH4/CH3OH, was used to improve sensitivity. The conversion of saccharide into alditol could reach nearly 100% in the NaBH4 reduction reaction. In addition, the bioanalytical quantitative screening method was validated according to US-FDA guidance, including selectivity, linearity, precision, accuracy, stability, and matrix effect. The developed method displayed a good accuracy, high sensitivity (LOD = 0.05 mg L−1), and good reproducibility (RSD < 15%). The assay accurately measured an IC50 value of 0.40 μM for the known AFU inhibitor, deoxyfuconojirimycin, which was consistent with results reported in the literature. Further validation of the assay was achieved through the determination of a high Z′-factor value of 0.89. The assay was applied to screen a marine-derived chemical library against AFU3, which revealed two marine-oriented pyrimidine alkaloids as potential AFU3 inhibitors. Graphical abstract [ABSTRACT FROM AUTHOR]

Deuterium-enriched and fluorine-substituted compounds have been widely applied in drug discovery due to their advantages in the studies of clinical pharmacokinetics and metabolic profiles. Herein we synthesized a library of deuterated and fluorine-substituted plinabulin derivatives, and all 15 D- or F-compounds were characterized by MS, $$^{1}\hbox {H}$$ NMR and $$^{ 13}\hbox {C}\,\hbox {NMR}$$ . Their antitumor activities were evaluated against human Jurkat T lymphocyte cells. [ABSTRACT FROM AUTHOR]

A hyaluronate lyase was obtained by cultivating Arthrobacter globiformis strain A152. The enzyme was purified to homogeneity from the supernatant by ammonium sulfate fractionation, Q Sepharose Fast Flow, and Sephadex G-100 chromatography. The purification resulted in a 32.78-fold increase in hyaluronate lyase activity with specific activity of 297.2 U/mg. The molecular weight of the enzyme determined by SDS-PAGE was approximately 73.7 kDa. Using hyaluronic acid (HA) as a substrate, the maximal reaction rate (V) and the Michaelis-Menten constant (K) of hyaluronate lyase were found to be 4.76 μmol/min/ml and 0.11 mg/ml, respectively. The optimum pH and temperature values for hyaluronate lyase activity were pH 6.0 and 42 °C, respectively. This enzyme was stable at pH 4-10, 5-7, and 5-7 at 4, 37, and 42 °C, respectively. Investigation about temperature effects on hyaluronate lyase displayed that it was stable at 30-37 °C and also showed high activity at 37 °C. The enzymatic activity was enhanced by Ca and was strongly inhibited by Cu and SDS. These properties suggested that the hyaluronate lyase in this study could bring promising prospects in medical and industry applications. [ABSTRACT FROM AUTHOR]

Plinabulin, a potent microtubule-targeting agent, is derived from marine natural diketopiperazine 'phenylahistin'. To develop novel plinabulin analogue that could display better pharmacokinetic properties and less side effects, deuterated plinabulin 9 was synthesized and evaluated in vitro and in vivo. In comparison with plinabulin, in vivo pharmacokinetic studies indicated that the deuterated derivative 9 could alter blood circulation behavior obviously, which was proved by increased area under the plasma concentration- time curve (AUC), reduced clearance (CL), and prolonged total body mean residence time (MRT). The derivative 9 also has higher inhibition rates against BxPC-3, Jurkat and A-431 tumor cell lines as compared with its prototype plinabulin. Therefore, the deuterated compound 9 might be developed as a potential agent for different cancer treatments. [ABSTRACT FROM AUTHOR]

The response surface methodology (RSM) combined with bioassays was employed to optimize the extraction process of crude fucose-containing sulphated polysaccharides (cFCSP) from Sargassum fusiforme. The central composite design (CCD) was used with four variables, five levels, and four responses. The four variables were pH value of hydrochloric acid solution, extraction temperature (°C), ratio of liquid to raw material (mL g), and extraction time (h), respectively. Chemical and bioassay indices were used in combination as the response parameters, which included the yield of cFCSP, fucose content, proliferation rate of spleen cells, and lipopolysaccharide-induced proliferation of splenocytes. The experimental data were fitted to a second-order polynomial equation using multiple regression analysis, and examined using the appropriate statistical methods. The best extraction conditions were as follows: the pH value of hydrochloric acid solution was 3.50; the extraction temperature was 100°C; the ratio of liquid to raw material was 15.00 mL g and the extraction time was 2.50 h. The experimental yield was close to the predicted from the model. The extract could promote spleen lymphocyte proliferation, especially the lipopolysaccharide-induced lymphocyte proliferation in vitro, which suggested that its immunomodulatory effect on B lymphocytes. Therefore, cFCSP extracted from S. fusiforme could be utilized as an immunostimulant in functional foods and pharmaceutical industry in future. [ABSTRACT FROM AUTHOR]

A rapid and sensitive fluorescence labeling method was developed and validated for the microanalysis of a sulfated polysaccharide drug,namely propylene glycol alginate sodium sulfate (PSS), in rat plasma. Fluorescein isothiocyanate (FITC) was selected to label PSS, and 1, 6-diaminohexane was used to link PSS and FITC in order to prepare FITC-labeled PSS (F-PSS) through a reductive amination reaction. F-PSS was identified by UV-Vis, FT-IR and H-NMR spectrum. The cell stability and cytotoxicity of F-PSS were tested in Madin-Darby canine kidney (MDCK) cells. The results indicated that the labeling efficiency of F-PSS was 0.522% ± 0.0248% and the absolute bioavailability was 8.39%. F-PSS was stable in MDCK cells without obvious cytotoxicity. The method was sensitive and reliable; it showed a good linearity, precision, recovery and stability. The FITC labeling method can be applied to investigating the absorption and metabolism of PSS and other polysaccharides in biological samples. [ABSTRACT FROM AUTHOR]

Tamarixetin and kaempferide, the major bioactive constituents of Xiheliu extract, have been simultaneously/quantitatively determined in rat plasma and urine by a sensitive high performance liquid chromatography method after oral administration the total flavonoids from Xiheliu. In this study, the biological samples were prepared by solid-phase extraction, then simultaneously detected at 254 nm and successfully separated and quantified using a reversed-phase C column with methanol-formic acid aqueous gradient solution, at a flow rate of 1 mL/min. Good linearity ( r > 0.989) of tamarixetin was observed in plasma and urine with the calibration ranges both at 1.6-80 μg/mL. For kaempferide, the correlation coefficient reached 0.994 in plasma at 1.4-70 μg/mL. The RSD of intra- and inter-day were 1.9-6.5% for tamarixetin and 1.3-9.0% for kaempferide in plasma; in urine, the intra- and inter-day RSD for not only tamarixetin but also kaempferide was no more than 7.4 and 5.8%, respectively. The lowest extraction recovery was 87.6% for kaempferide and 93.2% for tamarixetin in plasma and urine for both low and high concentrations. Due to the high sensitivity (the LOQ for tamarixetin was 1.2 μg/mL and for kaempferide 1.4 μg/mL), accuracy, precision, and good selectivity, the assay was successfully applied to pharmacokinetic studies of both flavonols in rats. The half-lives of tamarixetin and kaempferide were 17.8 ± 1.4 and 92.5 ± 1.6 min, and the c were 3.1 ± 0.2 and 2.5 ± 0.4 μg/mL, respectively. [ABSTRACT FROM AUTHOR]

It has been found that deep-sea water was associated with lower serum lipid in animal model studies. Herein, we investigated whether DSW exerted a hypolipidemic activity and further elucidated how DSW modulated lipid metabolism in HepG2 cells. Preliminary animal studies showed that DSW exhibited potency to decrease serum total cholesterol, triglycerides, and LDL cholesterol, and increase HDL cholesterol, and the hepatic lipid contents were also significantly lower in the DSW group. When DSW was added to HepG2 cells, it decreased the lipid contents of hepatocyte through the activation of AMP-activated protein kinase, thus inhibiting the synthesis of cholesterol and fatty acid. Besides, LDL receptor was upregulated by activation of sterol regulatory element-binding protein-2. In addition, the levels of apolipoprotein AI and cholesterol 7-alpha-hydroxylase were also raised. Our investigation provided mechanisms by which DSW modulated lipid metabolism and indicated that DSW was worthy of further investigation and could be developed as functional drinking water in the prevention and treatment of hypolipidemic and other lifestyle-related diseases. [ABSTRACT FROM AUTHOR]

In this study, seawater was pumped up from 150, 200, 300, 500 and 1000 m in the South China Sea and analyzed to make certain what depth should deep-sea water (DSW) be pumped up for medicinal usage. The pumping depth of DSW was determined on the basis of chemical ingredients. The analyses of inorganic elements and dissolved organic matter (DOM) were performed by inductively coupled plasma mass spectrometry (ICP-MS) and ultra performance liquid chromatography-mass spectrometry (UPLC-MS) respectively. The raw data were used for hierarchical cluster analysis (HCA) and principal component analysis (PCA). The results showed that seawater pumped up from 500 m and 1000 m was similar in their chemical ingredients, and was different from the seawater pumped up from other depths. These results indicated that seawater from more than 500 m depth had relatively stable chemical ingredients and could be used as DSW in the South China Sea. [ABSTRACT FROM AUTHOR]

n excess of reactive oxygen species (ROS) leads to a variety of chronic health problems. As potent antioxidants, marine bioactive extracts containing oligosaccharides and peptides have been extensively studied. Recently, there is a growing interest in protein-polysaccharide complexes because of their potential uses in pharmaceutical and food industries. However, only few studies are available on the antioxidant activities of such complexes, in terms of their ROS scavenging capability. In this study, we combined different marine oligosaccharides (isolated and purified) with collagen peptides derived from tilapia fish skin, and evaluated the antioxidant activity of the marine peptide-oligosaccharide complexes vis-à-vis the activity of their original component molecules. Biochemical and cellular assays were performed to measure the scavenging effects on 1, 1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl and superoxide radicals, and to evaluate the influences on the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the level of malondialdehyde (MDA) in UV-induced photoaging models. The results indicated that the antioxidant activities of all the complexes were stronger than those of their individual components. Among the 11 complexes tested, two complexes, namely MA1000+CP and κ-ca3000+CP, turned out to be highly effective antioxidants. Although the detailed mechanisms of this improved scavenging ability are not fully understood, this work provides insights into the design of highly efficient peptide-oligosaccharide complexes for potential applications in pharmaceutical, cosmetics and food industries. [ABSTRACT FROM AUTHOR]

Three kinds of polysaccharides: GFW, GFH and GFA, were sequentially extracted from a red alga Gloiopeltis furcata with 25°C and 85°C water, and 60 4% NaOH water solution. Based on the defatted alga, the yields of the polysaccharide were 57.9%, 2.5% and 2.6%, respectively. Their monosaccharide compositions, average molecular weights and structural characters were determined by gas chromatography (GC), high performance liquid chromatography (HPLC), fourier transform infrared spectroscopy (FTIR) or 13C-NMR spectroscopy. The results showed that GFW, GFH and GFA were all composed of D-galactose (Gal) and 3,6-anhydro-L-galactose (AnG), and particularly GFA also contained xylose (Xyl). The average molecular weights of GFW, GFH and GFA were 22.6 kD, 26.5 kD and 49.8 kD, respectively, with the respective sulfate content 31.2%, 25.1% and 22.7%. The data of FTIR and 13C-NMR confirmed the sulfate ester location at C6 of galactose. It is concluded that all the three polysaccharides extracted from Gloiopeltis furcata were sulfated galactans, two being sulfated-agarose, and one being xylose-containing sulfated galactan. [ABSTRACT FROM AUTHOR]

In order to determine the role of alginate-derived oligosaccharides (ADO) in drought stress resistance of tomato ( Lycopersicon esculentum Miller) seedlings, the leaves were exposed to different concentrations of ADO (0.05%, 0.10%, 0.20%, 0.30% and 0.50%) after drought stress was simulated by exposing the roots to 0.6 molL−1 PEG-6000 solution for 6 h. Changes in biomass, electrolyte leakage and malondialdehyde (MDA), free proline, total soluble sugars (TSS) and abscisic acid (ABA), the enzyme activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD) and phenylalanine ammonia-lyase (PAL) were measured to investigate the effects of ADO treatment. The results showed that the treatment with an ADO concentration of 0.20% exhibited the highest performance of drought stress resistance in the tomato seedlings by decreasing the electrolyte leakage and the concentration of MDA, increasing the contents of free proline, TSS and ABA, and increasing the activities of CAT, SOD, POD and PAL after treatment with ADO. It is suggested that changes in electrolyte leakage, MDA, osmotic solutes, ABA, anti-oxidative enzyme and PAL activities were responsible for the increased drought stress resistance in tomato seedlings. To our best knowledge, this is the first report of the effect of ADO treatment on enhancing the drought stress resistance of tomato seedlings. [ABSTRACT FROM AUTHOR]

Four typical coastal sites (rocky shore, sandy shore, mud flat shore, and artificial harbor) at the Yellow Sea were chosen to investigate the spatial and seasonal variations in bacterial communities. This was accomplished by using terminal restriction fragment length polymorphism (T-RFLP) analysis of PCR amplified 16S rDNA fragments. Two kinds of tetrameric restriction enzymes, HhaI and MspI, were used in the experiment to depict the bacterial community diversity in different marine environments. It was found that the community compositions digested by the two enzymes separately were different. However, the results of bacterial community diversity derived from them were similar. The MDA analysis results of T-RFLP profiles coming from HhaI and MspI both exhibited a significant seasonal community shift for bacteria and a relatively low spatial variation among the four locations. With HhaI as the sample, the pair wise T-tests also revealed that variations were minor between each pair of marine environments, with R ranging from 0.198 to 0.349. However, the bacterial community structure in the mud flat site depicted a larger difference than each of the other three sites ( R ranging from 0.282 to 0.349). [ABSTRACT FROM AUTHOR]

A high-performance liquid chromatographic method has been developed for the simultaneous analysis of the flavonols myricitrin ( 1), avicularin ( 2), and juglanin ( 3) in rat plasma and urine after oral administration of the total flavonoids from Polygonum aviculare. Samples were prepared by solid-phase extraction then separated on a C18 reversed-phase column by use of a mobile-phase gradient prepared from methanol and aqueous formic acid solution. The flow rate was 1 mL min−1. Detection was performed at 254 nm. The calibration range was 11–1,100 μg mL−1 for both 2 and 3 in plasma; in urine the calibration ranges for 1, 2, and 3 were 32–1,600, 11–1,100, and 22–1,100 μg mL−1, respectively. Intra-day and inter-day RSD were less than 4.33 and 3.62% for 2 and 3, respectively, in plasma, and no more than 4.03 and 2.22% for all the analytes in urine. The analytical sensitivity and selectivity of the assay enabled successful application to pharmacokinetic studies of flavonols 1– 3 in rats. [ABSTRACT FROM AUTHOR]

Confocal laser scanning microscopy (CLSM) was applied to detect the intracellular [Ca2+] variety of fluorescent intension, with Fluo-3/AM fluorescence loaded in SFSMC. The results show that 10 μmol/L Lacidipine can reduce the frequence which 10 μmol/L 5-HT induced [Ca2+] spark in SFSMC of calcium over loading to 50%, and amplitude to 50% or so. We can draw a conclusion that dihydropyridines calcium antagonists lacidipine can antagonize the release of intracellular [Ca2+] which 5-HT-induced in dose dependent manner. [ABSTRACT FROM AUTHOR]

Endogenous tubulin promoter has been widely used for expressing foreign genes in green algae, but the efficiency and feasibility of endogenous tubulin promoter in the economically important Porphyra yezoensis (Rhodophyta) are unknown. In this study, the flanking sequences of beta-tubulin gene from P. yezoensis were amplified and two transient expression vectors were constructed to determine their transcription promoting feasibility for foreign gene gusA. The testing vector pATubGUS was constructed by inserting 5′-and 3′-flanking regions ( Tub5′ and Tub3′) up-and down-stream of β-glucuronidase (GUS) gene ( gusA), respectively, into pA, a derivative of pCAT®3-enhancer vector. The control construct, pAGUSTub3, contains only gusA and Tub3′. These constructs were electroporated into P. yezoensis protoplasts and the GUS activities were quantitatively analyzed by spectrometry. The results demonstrated that gusA gene was efficiently expressed in P. yezoensis protoplasts under the regulation of 5′-flanking sequence of the beta-tubulin gene. More interestingly, the pATubGUS produced stronger GUS activity in P. yezoensis protoplasts when compared to the result from pBI221, in which the gusA gene was directed by a constitutive CaMV 35S promoter. The data suggest that the integration of P. yezoensis protoplast and its endogenous beta-tubulin flanking sequences is a potential novel system for foreign gene expression. [ABSTRACT FROM AUTHOR]

The influence of alginate-derived oligosaccharide (molecular weight 1445 Da) was tested on the maize seed germination at different concentrations. Assays of α-and β-amylase and protease activities showed the highest response at 0.75‰.Compared with the control, root growth on days 3 and 7 showed increases of 34% and18%, respectively; and shoot growth on day 7 an increase of 46%. In the case of protease activity, treatments with both 0.75‰ and 1.50‰ alginate-derived oligosaccharide gave higher activities than the control. These results indicate that the rate of seed germination was enhanced by increasing the activities of several enzymes beneficial for germination. [ABSTRACT FROM AUTHOR]

Depolymerization of κ-carrageenan was performed using carrageenase isolated from the cell-free medium of a culture of marine Cytophaga sp. MCA-2. The low-molecular-weight carrageenans after ultrafiltration and lyophilization were sulfonated with formamide-chlorosulfonic acid. The anti-tumor activity of the products with different molecular weight was determined by using Sarcoma 180 tumor in mouse. A carrageenan oligosaccharide with a molecular weight of 1726, administered orally at a dose of 100 mg kg−1 mouse markedly inhibited tumor formation. However, the anti-tumor activity of high-sulfonated carrageenan was much less than that of the non-sulfonated or light-sulfonated preparation. The activities of the latter products on superoxide dismutase and catalase were enhanced considerably, which suggests that carrageenan oligosaccharide was effective in promoting the antioxidation ability and eliminating danger from free radicals. The preparations showed special effects on immunological regulation, especially the phagocytosis ratio and phagocytosis index of macrophage, which might be beneficial for the anti-tumor activity. Although no anti-tumor activity of this product was detected in vitro, suggesting that its activity differs between in vitro and in vivo, this 1726 molecular weight product provides a potent clinical use in tumor treatment. [ABSTRACT FROM AUTHOR]