Abstract  The interfacial partitioning behavior of ampicillin and phenylglycine crystals in different two-phase systems has been investigated. The two-phase systems employed are water/dodecane, water/1-butanol, and water/pentane/methanol. By means of partition experiments and microscopic imaging, it has been shown that the mechanism of separation strongly depends on the choice of the two-phase system. While water/dodecane features a mechanism of sheer competitive adsorption at the interface, separation in water/1-butanol is mainly due to partitioning into both liquid phases, leading to a higher degree of separation. Experiments with water/pentane/methanol have illustrated the large potential of three-component systems, as slight variations in the composition can have large effects on the separation.

Abstract  Epstein-Barr virus (EBV) latent infection of B cells blocks the interrelated signaling and antigen-trafficking functions of the BCR through the activity of its latent membrane protein 2A (LMP2A). At present, the molecular mechanisms by which LMP2A exerts its control of BCR functions are only poorly understood. Earlier studies showed that in B cells expressing LMP2A containing a tyrosine mutation at position 112 in its cytoplasmic domain (Y112-LMP2A), the BCR could initiate signaling but could not properly traffic antigen for processing. Here, we show that BCR signaling in Y112-LMP2A-expressing cells is attenuated with a reduction in both the degree and duration of phosphorylation of key components of the BCR signaling cascade including Syk, BLNK, PI3K, and Btk. Notably, Y112-LMP2A expression completely blocked the BCR-induced activation of phospholipase D (PLD), a lipase implicated in the intracellular trafficking of a variety of surface receptors. We show that blocking PLD activity, by expressing Y112-LMP2A, treating cells with the PLD inhibitor 1-butanol or reducing PLD expression by siRNA, blocked BCR trafficking to class II-containing compartments. Moreover, Y112-LMP2A expression blocked the recruitment of phosphorylated forms of the downstream BCR signaling components, Erk and JNK, through both PLD-dependent and PLD-independent mechanisms. Thus, the investigation of the mechanism by which Y112-LMP2A blocks BCR function revealed an essential role for PLD in BCR trafficking for antigen processing.

Abstract  OBJECTIVES: To find the efficacy of serial extracts of Anisomeles malabarica in inhibiting proliferation of and inducing apoptosis in human cervical cancer cells, SiHa and ME 180, that are HPV 16-positive.

MATERIALS AND METHODS: The whole plant was extracted in n-hexane, chloroform, ethyl acetate, n-butanol, methanol, and water. The cells were treated with the extracts at increasing concentrations to find the IC(50), adopting MTT ([3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]) assay. Acridine orange (AO) and ethidium bromide (EB) and Hoechst 33258 staining were adopted to assess the mode of cell death, Annexin V-Cy3 staining to evaluate one of the early apoptotic features, JC-1 staining to assess the mitochondrial membrane depolarization, comet assay for DNA fragmentation, and cell cycle analysis for the distribution of cells after treatment.

RESULTS: n-Hexane and chloroform extracts were cytotoxic to the cervical cancer cells in dose- and duration-dependent manner. The cells that responded to the treatments revealed typical apoptotic features. Early features of apoptosis, phosphatidyl serine translocation and loss of mitochondrial trans-membrane potential, were observed in the treated cells, and comet assay revealed DNA damage. In the FACS analysis, the cells accumulated in the sub-G0/G1 phase of the cell cycle, except in n-hexane- and chloroform extract-treated SiHa cells at 24 h, which showed arrest in S- and G2/M phases.

CONCLUSIONS: n-Hexane and chloroform extracts of A. malabarica inhibit proliferation of and induce death in HPV16-positive cervical cancer cells, mostly by apoptosis and to some extent by necrosis.

Abstract  Canthaxanthin has a substantial commercial market in aquaculture, poultry production, and cosmetic and nutraceutical industries. Commercial production is dominated by chemical synthesis; however, changing consumer demands fuel research into the development of biotechnology processes. Highly productive microbial systems to produce carotenoids can be limited by the efficiency of extraction methods. Extraction with hexane, acetone, methanol, 2-propanol, ethanol, 1-butanol, tetrahydrofuran and ethyl acetate was carried out with each solvent separately, and subsequently the most efficient solvents were tested in combination, both as mixtures and sequentially. Sequential application of methanol followed by acetone proved most efficient. Extraction efficiency remained stable over a solvent to biomass range of 100:1 to 55:1, but declined significantly at a ratio of 25:1. Application of this method to a canthaxanthin-producing Escherichia coli production system enabled efficient canthaxanthin extraction of up to 8.5mgg(-1) dry biomass.

Abstract  The aim of this investigation was to evaluate the biological, alcohol dehydrogenase (ADH) and antiproliferative activities of different extracts of mungbean seeds and sprouts. All extracts from the sprouts showed higher contents of total phenolics (TP), total flavonoids (TF), and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity than from seeds. The highest DPPH and tyrosinase inhibition activities were registered in ethyl acetate (EtOAc) extract. ADH activity of methanol (MeOH), n-hexane (n-hexane) and n-butanol (n-BuOH) extracts from sprouts was significantly higher (P < 0.05) than from seeds. However, the highest ADH activity was found in water extract of seeds. According to 3-(4, 5-dimethylthiazol -2-yl)-2, 5-diphenyltetrazolium bromide) (MTT) assay, extracts from sprouts were more effective against Calu-6 (human pulmonary carcinoma) and SNU-601 (human gastric carcinoma) cells than from seeds. EtOAc extract showed the highest antiproliferative activity in both sprouts and seeds, followed by n-hexane extraction. During sprouting of mungbean, all the studied components significantly increased. In conclusion, the extracts of sprouts are more effective than from seeds and could be a potential source of antioxidants linked with health benefits.

Abstract  The nematode Caenorhabditis elegans arrests development at the first larval stage if food is not present upon hatching. Larvae in this stage provide an excellent model for studying stress responses during development. We found that supplementing starved larvae with ethanol markedly extends their lifespan within this L1 diapause. The effects of ethanol-induced lifespan extension can be observed when the ethanol is added to the medium at any time between 0 and 10 days after hatching. The lowest ethanol concentration that extended lifespan was 1 mM (0.005%); higher concentrations to 68 mM (0.4%) did not result in increased survival. In spite of their extended survival, larvae did not progress to the L2 stage. Supplementing starved cultures with n-propanol and n-butanol also extended lifespan, but methanol and isopropanol had no measurable effect. Mass spectrometry analysis of nematode fatty acids and amino acids revealed that L1 larvae can incorporate atoms from ethanol into both types of molecules. Based on these data, we suggest that ethanol supplementation may extend the lifespan of L1 larvae by either serving as a carbon and energy source and/or by inducing a stress response.

Abstract  Salvia sclareoides is an aromatic herb native to Portugal, of which phenolic content (Folin-Ciocalteau method), chemical profile (HPLC/DAD), antioxidant activity (DPPH, beta -carotene/linoleic acid assays), acute toxicity (MTT method, adapted for non-adherent cells), genotoxicity (short-term chromosomal aberration assay) and prion binding properties were evaluated in the acetone, water, ethanol, methanol and n-butanol extracts. The latter presented the highest phenolic content and antioxidant activity (DPPH assay), and was the single one with the flavonoids (+)-catechin, kaempferol O-glucoside and quercetin. Vanillic acid was the major component of all extracts but gallic, gentisic, caffeic, syringic, coumaric and ferulic acids were also found in some extracts. Only the n-butanol extract had components binding to the cellular form of human prion protein detected by NMR which showed specificity for two regions of the folded domain and for the unstructured N-terminal region. Extracts were not cytotoxic nor genotoxic, reinforcing the potential of S. sclareoides for nutraceutical purposes.

Abstract  Lespedeza cuneata G. Don is a plant commonly grown in Asian countries, which has been widely used as an oriental medicinal herb to treat diabetes, diarrhea and various other inflammatory diseases. The phenolics of dry leaves of L. cuneata G. Don were extracted by using 80% (v/v) aqueous methanol in assistance with homogenization and sonification. The phenolic extract and its five different fractions (n-hexane, chloroform, ethyl acetate, n-butanol, and water) were used to evaluate the levels of total phenolics, total flavonoids, and antioxidant capacity as well as the inhibitory effect of tyrosinase activity. Ethyl acetate fraction (1 g) had the highest levels of total phenolics at 240.8 mg gallic acid equivalents (GAF), total flavonoids as 90.4 mg catechin equivalents (CE) as well as antioxidant capacity at 523.4 mg vitamin C equivalents (VCE) on ABTS assay and 329.5 mg VCE on DPPH assay among fractions. One g of water fraction contained total phenolics at 133.1 mg GAE, total flavonoids at 34.5 mg CE, and antioxidant capacity at 333.4 mg VCE for ABTS assay and 313.2 mg VCE for DPPH assay. inhibition of tyrosinase activity of water fraction at 300 mu g.mL(-1) was at 47.2% and 21.1 % for L-tyrosine and L-DOPA as its substrate, respectively. On the other hand, ethyl acetate fraction at 300 mu g.mL(-1) showed tyrosinase inhibition of 10.2% for L-tyrosine and 11.9% for L-DOPA. These results suggested that the phenolics from dry leaves of L. cuneata G. Don may be utilized as a potent source of antioxidants and skin whitening agents.

Abstract  OBJECTIVES: The aim of this study was to determine if spray-drying could successfully produce microparticles containing the model protein trypsin in a form suitable for inhalation.

METHODS: Trypsin was spray-dried with raffinose from a methanol : n-butyl acetate solvent system (MeOH : BA). The solvent system was then adjusted to include water, and trypsin was co-spray-dried with raffinose, trehalose or hydroxpropyl-β-cyclodextrin. The spray-dried products were characterised by SEM, XRD, DSC, TGA and FTIR. Protein biological activity and in-vitro deposition of trypsin : excipient nanoporous/nanoparticulate microparticles (NPMPs) was also assessed.

KEY FINDINGS: The inclusion of water in a MeOH : BA solvent system allowed for the successful production of NPMPs of trypsin : excipient by spray-drying. Trypsin formulated as trypsin : excipient NPMPs retained biological activity on processing and showed no deterioration in activity or morphological characteristics when stored with desiccant at either 4 or 25°C. Hydroxpropyl-β-cyclodextrin showed advantages over the sugars in terms of producing powders with appropriate density and with greater physical stability under high-humidity conditions. Fine particle fractions of between 41 and 45% were determined for trypsin : excipient NPMPs.

CONCLUSIONS: NPMPs of trypsin : excipient systems can be produced by spray-drying by adjustment of the solvent system to allow for adequate solubility of trypsin.

Abstract  A chemiresistor type sensor for selective alcohol sensing has been realized from gallium nitride (GaN) nanowires (NWs) functionalized with sputter-deposited tin dioxide (SnO2) nanoparticles. Two-terminal devices were fabricated using standard microfabrication techniques with the individual NWs air-bridged between the two metal contact pads. Through a combination of X-ray diffraction (XRD), electron-backscatter-diffraction (EBSD), and TEM/STEM techniques, we confirmed the presence of rutile SnO2 nanocrystals on the GaN surface. A change in device current is observed when the device is exposed to alcohol vapors (methanol, ethanol, propanol, and butanol) at room temperature under 215-400 nm UV illumination with 3.75 mW/m(2) intensity at 365 nm wavelength. The sensor reproducibly responded to a wide range of alcohol vapor concentrations, from 5000 mu mol/mol (ppm) down to 1 mu mol/mol (ppm) in air. Notably, the devices show low sensitivity to acetone and hexane, which allows them to selectively detect the primary alcohol vapors mixed with these two common volatile organic compounds (VOCs). The sensor response was not observed without UV excitation. From the experimental results we found a relationship between the response towards the alcohol vapors and the length of carbon chain in the molecule: the chemiresistive response decreases with the increasing carbon chain from methanol to n-butanol. In addition, we observed that the isomeric branching in i-propanol and i-butanol caused reduced response as compared to n-propanol and n-butanol, respectively. We have qualitatively explained the sensor operation by employing a mechanism, which includes oxidation of analyte molecules on the SnO2 surface, leading to enhanced photoconductivity in GaN nanowire. (C) 2012 Elsevier B.V. All rights reserved.

Abstract  The Present study was conducted to investigate the biological activities of the plant O. monacantha cladodes. The plant cladodes were extracted with methanol and then fractionized with various organic solvents (n-hexane, chloroform, ethyl acetate and n-butanol). The extract and fractions thus obtained were analyzed for their phytochemical, antioxidant and antimicrobial analysis. The percentage yield of methanolic extract and various fractions was found to be 2.7-28.9%. Phytochemical analysis exposed the presence of alkaloids, tannins, saponins and flavonoids in methanolic extract. The plant extract and fractions contained total phenolic contents (14.5-249 GAE, mg/100 g of dry plant matter) and total flavonoid contents (10-433.8 CE, mg/100 g of dry plant matter). DPPH radical scavenging activity was determined by measuring IC50 that was found to be 41.4-838.8 mu g/mL. Nevertheless, minute variation was observed in reducing power. Methanol extract exhibited highest antioxidant activity among various solvent fractions. The antimicrobial activity was determined by disc diffusion method and minimum inhibitory concentration against a panel of microorganisms. Significant variations (p < 0.05) were observed in the results.

Abstract  The present study aimed to investigate the direct in vitro effects of several distinct Citrullus colocynthis seed extracts on glucose-stimulated insulin release from pancreatic islets isolated from rats. Six extracts were tested, a crude aqueous, defatted aqueous, ethyl acetate, H2O-methanol and n-butanol extract and an extract containing a major component (fraction A) identified by gel chromatography in the ethyl acetate, n-butanol and H2O-methanol extracts. Under selected experimental conditions, the majority of extracts exhibited a positive insulinotropic action, at least when tested in the presence of 8.3 mM D-glucose. The concentration-response correlation observed with distinct extracts revealed the participation of distinct chemical compounds, including compounds with an inhibitory insulinotropic potential, in the modulation of the insulin secretory response to D-glucose. The results of the present study are relevant for further investigations which aim to identify compounds exhibiting positive insulinotropic actions. These agents may be suitable for the treatment of human diabetic subjects.

Abstract  Butanol is a very competitive renewable biofuel for use in internal combustion engines given its many advantages. In this review, the properties of butanol are compared with the conventional gasoline, diesel fuel, and some widely used biofuels, i.e. methanol, ethanol, biodiesel. The comparison of fuel properties indicates that n-butanol has the potential to overcome the drawbacks brought by low-carbon alcohols or biodiesel. Then, the development of butanol production is reviewed and various methods for increasing fermentative butanol production are introduced in detailed, i.e. metabolic engineering of the Clostridia, advanced fermentation technique. The most costive part of the fermentation is the substrate, so methods involved in renewed substrates are also mentioned. Next, the applications of butanol as a biofuel are summarized from three aspects: (1) fundamental combustion experiments in some well-defined burning reactors; (2) a substitute for gasoline in spark ignition engine; (3) a substitute for diesel fuel in compression ignition engine. These studies demonstrate that butanol, as a potential second generation biofuel, is a better alternative for the gasoline or diesel fuel, from the viewpoints of combustion characteristics, engine performance, and exhaust emissions. However, butanol has not been intensively studied when compared to ethanol or biodiesel, for which considerable numbers of reports are available. Finally, some challenges and future research directions are outlined in the last section of this review.

Abstract  The use and search for antibiotics and dietary supplements derived from plants have accelerated in recent years. Three plants, used traditionally as medicine and as food additive in Saudi Arabia, were collected and extracted with either methanol or n-butanol. The used plants were Rheum palmatum, Curcuma longa and Alpinia officinarum. The plant extracts were screened for their inhibitory effects on seven bacterial and five fungal genera using agar well diffusion method. It was shown that methanol extract was more effective as compared to n-butanol extracts. The minimum inhibitory concentrations (MIC) of the methanol extracts of the used plants ranged from 50 to 175 mu g/ml. No toxicity was found using Artimia salina as test organism. Antitumor activity against Ehrlich ascites carcinoma was recorded only for C. longa extract.

Abstract  BACKGROUND: Comprehensive kinetic models of microbial metabolism can enhance the understanding of system dynamics and regulatory mechanisms, which is helpful in optimizing microbial production of industrial chemicals. Clostridium acetobutylicum produces solvents (acetone-butanol-ethanol, ABE) through the ABE pathway. To systematically assess the potential of increased production of solvents, kinetic modeling has been applied to analyze the dynamics of this pathway and make predictive simulations. Up to date, only one kinetic model for C. acetobutylicum supported by experiment has been reported as far as we know. But this model did not integrate the metabolic regulatory effects of transcriptional control and other complex factors. It also left out the information of some key intermediates (e.g. butyryl-phosphate).

RESULTS: We have developed an improved kinetic model featured with the incorporation of butyryl-phosphate, inclusion of net effects of complex metabolic regulations, and quantification of endogenous enzyme activity variations caused by these regulations. The simulation results of our model are more consistent with published experimental data than the previous model, especially in terms of reflecting the kinetics of butyryl-phosphate and butyrate. Through parameter perturbation analysis, it was found that butyrate kinase has large and positive influence on butanol production while CoA transferase has negative effect on butanol production, suggesting that butyrate kinase has more efficiency in converting butyrate to butanol than CoA transferase.

CONCLUSIONS: Our improved kinetic model of the ABE process has more capacity in approaching real circumstances, providing much more insight in the regulatory mechanisms and potential key points for optimization of solvent productions. Moreover, the modeling strategy can be extended to other biological processes.

Abstract  Muscodor sutura is described as a novel species that is also an endophyte of Prestonia trifidi. Uniquely, this fungus produces a reddish pigment, on potato dextrose agar (PDA), when grown in the dark. In addition, the organism makes some volatile organic compounds that have not been previously reported from this genus, namely, thujopsene, chamigrene, isocaryophyllene, and butanoic acid, 2-methyl. These and other volatile compounds in the mixture possess wide-spectrum antifungal activity and no observable antibacterial activity. Most unusually, on PDA, the newly developing hyphae of this fungus grow in a perfect stitching pattern, in and out of the agar surface. The partial ITS-DNA sequence of this organism is identical to that of Muscodor vitigenus but it differs from all other Muscodor spp. Justification for a new species, as Muscodor sutura, is collectively based on morphological, cultural, chemical, and bioactivity properties.

Abstract  Artemisinin extracted from Artemisia annua L. is the best medicine with the highest efficiency, the most effective and the lowest toxicity in treating ague nowadays. At present, most studies focus on artemisinin and its derivatives, while the study and report about the other active components are rare. This paper purposed to further discover new indication of Artemisia annua L. connecting with the record of traditional medicine. We screened the hemostatic active fraction of Artemisia annua L. in-vitro by plasma recalcification time (PRT). The crude extract and the extract of n-butanol were purified by polyannde, MCI, gel column in order. Determining the part of 20% methanol fraction after column chromatography of MCI gel is the hemostatic active fraction of Artemisia annua L. The shorten rate of clotting time are followed by: crude extract of Artemisin annua L. (8.51%); the n-butanol extract (14.89%); water eluting fraction after the extract of n-butanol was purificated byipolyamide (22.11%); 20% methanol fraction after column chromatography of MCI gel (27.37%). It can provide experimental data for the clinical alication of Artemisia annua L. which can be exploited as hemostatic. This topic has a certain academic value and potential prospects on the deep research of the Artemisia annua L. resource.

Abstract  Aerial parts of Saussurea salicifolia (L.) DC were studied for their lignan and flavonoids in solvent chloroform and n-butanol of ethanolic extract. Isolation and identification of phenolic compounds of the chloroform and n-butanol fractions were performed with Dionex HPLC-DAD system with water-methanol gradients in 4 different wave lengths (235 nm, 254 nm, 280 nm and 340 nm), using online UV and LC-MS as described previously. 9-OH-pinoresinol which is a lignan with anticancer activity was dominated in the chloroform fraction, whereas mainly flavonoid glycosides like quercetin-3-O-galactoside, apigenin-7-O-rhamnoside with anti-inflammatory effect were detected in the n-butanol fraction. Additionally, 9-OH-pinoresinol was also found in the n-butanol fraction. Anticancer tests were conducted in leukemia mouse lymphoma cells L5178Y at a concentration of 10 mu g/ml of test compound. Crude ethanol extract of S. salicifolia reduced the growth of leukemia mouse lymphoma cells L5178Y to 23.8%.

Abstract  Extracts of medicinal plants have been reported to be very much effective in the control of chronic diseases due to their high antioxidant contents. Focus of the present study is the exploitation of safer and cheaper sources of the antioxidants based on the natural origin. Litichi chinensis Sonn. is used for its hypoglycemic activity in the traditional medicinal system of Indian subcontinent. The antioxidant activity of the aqueous and organic extracts of leaves, stem and fruit pulp of L chinensis was investigated using 2,2'-azinobis(3-ethylbenzothiazoline-6-sulpohonic acid) (ABTS) decolourization assay, the ferric reducing anti-oxidant power (FRAP) assay, 2.2'-diphenyl-1-picrylhydrazil (DPPH) assay, the total phenolic content (TPC) assay and the total antioxidant activity assay. Using the ABTS decolourization assay, L chinensis extracts showed a wide range of antioxidant activity in terms of trolox equivalent antioxidant capacity (TEAC); from 5.64-0.126 mM for leaves of L. chinensis (LCL), from 4.12-0.23 mM for stem (LCS) and from 0.66-0.24 mM for fruit pulp of L chinensis (LCPL). The total phenolic content assay exhibited 1.72, 1.46 and 1.41 mM gallic acid equivalents for ethyl acetate, aqueous (before partitioning) and aqueous (after partitioning) fractions of the stem respectively. The ferric reducing antioxidant power assay indicated high reducing power of leaves and stem of Litchi chinensis. Methanol, 1-butanol, aqueous and ethyl acetate fractions of all the parts of Litchi chinensis showed strong DPPH and peroxyl radicals scavenging activity. The results obtained in the present study demonstrated that all the pats studied are potential sources of natural antioxidants. Although the radical scavenging and inhibition of lipid peroxidation studies show usefulness of Litchi chinensis extracts in vitro conditions, yet verification of their in vivo activity and especially during disease conditions needs further experimentation.

Abstract  Conventional binder systems for tape casting contain toxic phthalate plasticizers and butanone (MEK) as part of the solvent. The effects of exchanging the phthalate with a non-toxic alternative, and butanone with ethanol, were studied on laminates of high-green density CGO (Ce0.9Gd0.1O1.95) tapes. Samples were prepared with a binder system containing DBP (dibutyl phthalate) plasticizer and MEK solvent, and with a binder system based on a non-toxic non-phthalate plasticizer and ethanol. In both systems, the weight ratio of plasticizer to the PVB (polyvinyl butyral) binder was varied between 0.4 and 0.7. Substitution to the less toxic binder system had no adverse impacts on the microstructure. In fact, denser packing and improved homogeneity were observed with the non-phthalate-based system at ratio 0.5 indicating improved dispersion in this system. The denser packing also coincided with a maximum in z-shrinkage and molecular weight of the binder system, which could be related to the distribution of the binder system. (C) 2009 Elsevier Ltd and Techna Group S.r.l. All rights reserved.

Abstract  PURPOSE: Asteroid hyalosis is a condition in which white spherical particles (asteroid bodies) are suspended in the vitreous, usually in the dependent part of the vitreous. These particles seldom cause serious visual symptoms; however, their presence can be a source of irritation. It has been suggested, but not confirmed, that asteroid hyalosis may be associated with systemic diseases such as diabetes, hyperlipidemia, or hypertension. Studies indicate that these particles are composed of lipid material and calcium; however, the specific composition and structure of asteroid bodies remains unknown. We have observed that asteroid hyalosis occurs in galactose-fed dogs, and this represents the first animal model which consistently forms this vitreal condition. The purpose of this study was to identify the main structural component of the asteroid bodies present in the vitreous of these dogs.

METHODS: Vitreous humor containing asteroid bodies was collected and frozen from long-term galactose-fed beagles and from age-matched normal controls where asteroid bodies were absent. A portion of the frozen vitreous was sent out for elemental analysis. Thawed vitreous samples were sonicated with HPLC grade water and the aqueous layer was extracted three times with chloroform and then three times with n-butanol. The three organic layers from each extraction were combined and the solvents removed in vacuo. The residue from each extraction was re-dissolved in methanol and analyzed by electrospray ionization mass spectrometry (ESI-MS).

RESULTS: Vitreous-containing asteroid bodies had significantly higher levels of calcium and phosphorus. Negative mode ESI-MS analysis of the n-butanol extracts from vitreous samples with and without asteroid bodies were similar with both containing a predominant peak with a mass to charge ratio (m/z) of 538.4. However, similar analyses of the chloroform extracts indicated that three peaks with m/z values of 547.1, 690.5, and 1430.6 were present only in vitreous samples containing asteroid bodies. Subtraction analysis indicated that the m/z of 690.5 peak corresponded to the main component present. This peak was identified and confirmed to be the quasimolecular ion of 1,2-dipalmitoyl-glycero-3-phosphoethanolamine (DPPE).

CONCLUSIONS: Based on the current belief that asteroid bodies are composed of lipid-calcium complexes, we propose that the main component of asteroid hyalosis in the galactose-fed dog is a quasimolecular ion of DPPE in which two molecules of DPPE are complexed through their phosphates groups with calcium.

Abstract  To understand antitumor activity of Albizzia julibrissin Durazz (Leguminosae), which has been used as a traditional oriental medicine, the mechanism underlying cytotoxic effect of its extract on human acute leukemia Jurkat T cells were investigated. The methanol extract of the stripped barks (3kg) of Albizzia julibrissin was evaporated, dissolved in water, and then sequentially extracted by chloroform, ethyl acetate, and n-butanol. The substance in the butanol extract containing the most cytotoxic activity was further purified by a series of preparative column chromatography. The active substance obtained (723mg) was designated as HaBC18. When Jurkat T cells were treated with HaBC18 (0.5-2microg/ml), apoptosis along with several biochemical events such as mitochondrial cytochrome c release, activation of caspase-9 and -3, degradation of PARP, and DNA fragmentation was induced in a dose-dependent manner. However, the HaBC18-induced apoptosis was abrogated by an ectopic overexpression of Bcl-xL, which is known to block mitochondrial cytochrome c release. Primary cultures of human PBMC were less sensitive to the cytotoxicity relative to Jurkat T cells. These results demonstrate that the cytotoxicity of HaBC18 toward Jurkat T cells is attributable to apoptosis mediated by mitochondria-dependent death-signaling pathway regulated by Bcl-xL.

Abstract  A new liquid chromatography (LC)-negative ion electrospray ionization (ESI(-))-tandem mass spectrometry (MS/MS) method with post-column addition of ammonia in methanol has been developed for the analysis of acid herbicides: 2,4-dichlorophenoxy acetic acid, 4-chloro-o-tolyloxyacetic acid, 2-(2-methyl-4-chlorophenoxy)butyric acid, mecoprop, dichlorprop, 4-(2,4-dichlorophenoxy) butyric acid, 2,4,5-trichlorophenoxy propionic acid, dicamba and bromoxynil, along with their degradation products: 4-chloro-2-methylphenol, 2,4-dichlorophenol, 2,4,5-trichlorophenol and 3,5-dibromo-4-hydroxybenzoic acid. The samples were extracted from the surface water matrix using solid-phase extraction (SPE) with a polymeric sorbent and analyzed with LC ESI(-) with selected reaction monitoring (SRM) using a three-point confirmation approach. Chromatography was performed on a Zorbax Eclipse XDB-C18 (50 x 4.6 mm i.d., 1.8 mum) with a gradient elution using water-methanol with 2 mM ammonium acetate mobile phase at a flow rate of 0.15 mL/min. Ammonia in methanol (0.8 M) was added post-column at a flow rate of 0.05 mL/min to enhance ionization of the degradation products in the MS source. One SRM transition was used for quantitative analysis while the second SRM along with the ratio of SRM1/SRM2 within the relative standard deviation determined by standards for each individual pesticide and retention time match were used for confirmation. The standard deviation of ratio of SRM1/SRM2 obtained from standards run on the day of analysis for different phenoxyacid herbicides ranged from 3.9 to 18.5%. Limits of detection (LOD) were between 1 and 15 ng L(-1) and method detection limits (MDL) with strict criteria requiring <25% deviation of peak area from best-fit line for both SRM1 and SRM2 ranged from 5 to 10 ng L(-1) for acid ingredients (except dicamba at 30 ng L(-1)) and from 2 to 30 ng L(-1) for degradation products. The SPE-LC-ESI(-) MS/MS method permitted low nanogram-per-liter determination of pesticides and degradation products for surface water samples.