Abstract  The antiproliferative activities of n-hexane, chloroform, aqueous-methanol and aqueous extracts of the aerial parts of the Achillea millefolium aggregate on three human tumour cell lines were investigated by means of MTT assays. The chloroform-soluble extract exerted high tumour cell proliferation inhibitory activities on HeLa and MCF-7 cells, and a moderate effect on A431 cells; accordingly, it was subjected to detailed bioactivity-guided fractionation. As a result of the multistep chromatographic purifications (VLC, CPC, PLC, gel filtration), five flavonoids (apigenin, luteolin, centaureidin, casticin and artemetin) and five sesquiterpenoids (paulitin, isopaulitin, psilostachyin C, desacetylmatricarin and sintenin) were isolated and identified by spectroscopic methods. The antiproliferative assay demonstrated that centaureidin is the most effective constituent of the aerial parts of yarrow: high cell growth inhibitory activities were observed especially on HeLa (IC(50) 0.0819 microm) and MCF-7 (IC(50) 0.1250 microm) cells. Casticin and paulitin were also highly effective against all three tumour cell lines (IC(50) 1.286-4.76 microm), while apigenin, luteolin and isopaulitin proved to be moderately active (IC(50) 6.95-32.88 microm). Artemetin, psilostachyin C, desacetylmatricarin and sintenin did not display antiproliferative effects against these cell lines. This is the first report on the occurrence of seco-pseudoguaianolides (paulitin, isopaulitin and psilostachyin C) in the Achillea genus.

Abstract  A simple, precise, accurate and rapid high performance thin layer chromatographic method has been developed and validated for the simultaneous estimation of valsartan and hydrochlorothiazide in combined dosage forms. The stationary phase used was precoated silica gel 60F(254). The mobile phase used was a mixture of chloroform: methanol: toluene: glacial acetic acid (6:2:1:0.1 v/v/v/v). The detection of spots were carried out at 260 nm. The method was validated in terms of linearity, accuracy, precision and specificity. The calibration curve was found to be linear between 300 to 800 ng/spot for valsartan and 100 to 600 ng/spot for hydrochlorothiazide. The limit of detection and the limit of quantification for the valsartan were found to be 100 and 300 ng/spot respectively and for hydrochlorothiazide 30 and 100 ng/spot respectively. The proposed method can be successfully used to determine the drug content of marketed formulation.

Abstract  Ailanthus excelsa (Roxb), an Egyptian medicinal species highly important for treating numerous diseases, was investigated against experimentally induced gastric ulcer in rodents. We evaluated the gastroprotective effect of four extracts (petroleum ether, diethyl ether, chloroform, and methanol) of A. excelsa bark by using the ethanol-induced gastric lesion model. The pretreatment of animals with methanolic, petroleum ether, and chloroformic extracts (100 mg/kg, oral (p.o.)) from A. excelsa significantly reduced gastric lesion induced by ulcerogenic agent (56, 47, and 70%, respectively) when compared with animals pretreated with vehicle. However, the diethyl ether pretreatment led to the least gastric lesion damage (83%), similar to the standard antiulcer drug, cimetidine, at the same dose (100 mg/kg, p.o.). The lower effective dose of diethyl ether extract, as well as cimetidine, given by intraduodenal route, significantly increased the pH values and reduced the acid output of gastric juice. Sterols, triterpenes,and quassinoids are present in the diethyl ether extract of A. excelsa stem bark, which presented the best gastroprotective action among the studied extracts. Our study confirmed the traditional indications of A. excelsa for the treatment of gastric ulcer.

Abstract  PURPOSE: The purpose of this work was to develop an enzymatic method of quantification of cholesterol and cholesterol esters derived from contact lenses, both in vitro and ex vivo.

METHODS: Lotrafilcon B (O2 Optix; CIBA Vision, Inc., Duluth, GA) and galyfilcon A (Acuvue Advance; Vistakon, Inc., Jacksonville, FL) silicone hydrogel contact lenses were independently incubated in cholesterol oleate solutions varying in concentrations. After incubation, the lenses were removed and underwent two separate 2:1 chloroform-methanol extractions. After in vitro studies, 10 human subjects wore both lotrafilcon B and galyfilcon A contact lenses for 7 days. The lenses also underwent two separate 2:1 chloroform-methanol extractions. All in vitro and ex vivo samples were quantified with a cholesterol esterase enzymatic reaction.

RESULTS: Calibration curves from quantifications of in vitro contact lens samples soaked in successively decreasing concentrations of cholesterol oleate yielded coefficients of determination (R(2)) of 0.99 (lotrafilcon B) and 0.97 (galyfilcon A). For in vitro contact lens samples, galyfilcon A was associated with an average cholesterol oleate extraction of 39.85 +/- 48.65 microg/lens, whereas lotrafilcon B was associated with 5.86 +/- 3.36 microg/lens (P = 0.05) across both extractions and all incubation concentrations. For ex vivo contact lens samples, there was significantly more cholesterol and cholesterol esters deposited on galyfilcon A (5.77 +/- 1.87 microg/lens) than on lotrafilcon B (2.03 +/- 1.62 microg/lens; P = 0.0005).

CONCLUSIONS: This is an efficient and simple method of quantifying total cholesterol extracted from silicone hydrogel contact lenses and, potentially, the meibum and/or tear film. Certain silicone hydrogel materials demonstrate more affinity for cholesterol and its esters than do others.

Abstract  ETHNOPHARMACOLOGICAL RELEVANCE: Copaifera multijuga Hayne (Leguminosae) is a tree that produces an oleoresin, which is extensively commercialized in Brazil as capsules or crude oil for the treatment of several disorders. Ethnopharmacological studies show a diversity of indications such as anti-inflammatory and epidermal wound cicatrization.

AIM OF THE STUDY: In the present work three fractions obtained from Copaifera multijuga oleoresin (hexane (HF), chloroform (CF), and methanol (MF) from a KOH impregnated silica gel column chromatography, representing the three main classes of compounds in the Copaifera genus (hydrocarbon sesquiterpenes, oxygenated sesquiterpenes and acidic diterpenes), were evaluated using antinociceptive and anti-inflammatory models.

MATERIALS AND METHODS: HF, CF, and MF (doses ranging between 1 and 150 mg/kg, depending on the model used), Copaifera multijuga oleoresin (CMO, 100mg/kg, p.o.) and the reference drug morphine (5mg/kg, p.o.) were evaluated using models for analgesia (acetic acid-induced contortions and tail flick) or inflammation (rat paw oedema and increase in vascular permeability). To elucidate the mechanism of action from the fractions, animals were pre-treated with naloxone (opioid receptor antagonist, 5mg/kg, i.p.).

RESULTS: Fractions significantly inhibited (in a concentration-dependant way) the number of contortions induced by acetic acid and the second phase of formalin-induced licking response. Similar results were observed in the tail flick model. The central antinociceptive effect for HF and CF at the doses of 50 and 100mg/kg was higher than the one observed for morphine (1mg/kg). Administration of naloxone inhibited the antinociceptive effect of fractions indicating that HF, CF, and MF may be acting on opioid receptors. All three fractions also inhibited rat paw oedema and the increase in vascular permeability induced by several phlogistic agents (carrageenan, histamine, and serotonin).

CONCLUSIONS: Our results indicate that fractions obtained from Copaifera multijuga Hayne demonstrate an antinociceptive effect probably mediated by opioid receptors, and anti-inflammatory activity through inhibition of histaminergic and serotoninergic pathways.

Abstract  Bradykinin (BK) is an active peptide that binds to the kinin B(2) receptor and induces biological events during the development and adult life. In this study we aimed to investigate the effect of kinin B(2) receptor ablation in the postnatal skeletal muscle development and body composition in adult life. For studies of skeletal muscle development, control (C57Bl6 - WT) and B(2) receptor knockout mice (B(2)(-/-)) were sacrificed at 15, 30 and 90days after birth, the gastrocnemius skeletal muscle was weighed and myostatin gene expression evaluated by real time PCR. For energy balance determination, data from control and B(2)(-/-) at 90 and 120days were collected by calorimetric method. Body composition at 120days was determined by chloroform-methanol (total body fat) and Lowry-modified method (total body protein). The results show that B(2)(-/-) have significantly increased total body weight at 15, 30 and 90days of life, when compared to WT. The weight of the gastrocnemius skeletal muscle was also significantly increased at 30 and 90days of life. Body composition analyses revealed that B(2)(-/-) mice exhibit more total corporal protein and less total corporal fat. Energy balance revealed that B(2)(-/-) have increased metabolizable energy intake and energy expenditure when compared to control mice, resulting in a lower energy gain. Interestingly, myostatin mRNA expression was significantly decreased in 15 and 30days old B(2)(-/-) mice and after icatibant treatment of WT adult mice for 5days. In conclusion, together our results show that kinin B(2) receptor deletion increases lean mass, reduces fat mass and improves metabolism efficiency in mice. The mechanism involved in this phenotype could be related to the reduction of myostatin gene expression during postnatal life.

Abstract  Trypsin/ethylenediaminetetraacetic acid (EDTA) treatment and cell scraping in a buffer solution were compared for harvesting adherently growing mammalian SW480 cells for metabolomics studies. In addition, direct scraping with a solvent was tested. Trypsinated and scraped cell pellets were extracted using seven different extraction protocols including pure methanol, methanol/water, pure acetone, acetone/water, methanol/chloroform/water, methanol/isopropanol/water, and acid-base methanol. The extracts were analyzed by GC-MS after methoximation/silylation and derivatization with propyl chloroformate, respectively. The metabolic fingerprints were compared and 25 selected metabolites including amino acids and intermediates of energy metabolism were quantitatively determined. Moreover, the influence of freeze/thaw cycles, ultrasonication and homogenization using ceramic beads on extraction yield was tested. Pure acetone yielded the lowest extraction efficiency while methanol, methanol/water, methanol/isopropanol/water, and acid-base methanol recovered similar metabolite amounts with good reproducibility. Based on overall performance, methanol/water was chosen as a suitable extraction solvent. Repeated freeze/thaw cycles, ultrasonication and homogenization did not improve overall metabolite yield of the methanol/water extraction. Trypsin/EDTA treatment caused substantial metabolite leakage proving it inadequate for metabolomics studies. Gentle scraping of the cells in a buffer solution and subsequent extraction with methanol/water resulted on average in a sevenfold lower recovery of quantified metabolites compared with direct scraping using methanol/water, making the latter one the method of choice to harvest and extract metabolites from adherently growing mammalian SW480 cells.

Abstract  The antiproliferative effects of n-hexane, chloroform and aqueous methanol extracts prepared from the whole plant of Centaurea arenaria M.B. ex Willd. were investigated against cervix adenocarcinoma (HeLa), breast adenocarcinoma (MCF7) and skin epidermoid carcinoma (A431) cells, using the MTT assay. The chloroform extract displayed high tumour cell proliferation inhibitory activity (higher than 85% at 10 μg/mL concentration), and was therefore subjected to a bioassay-guided multistep separation procedure. Flavonoids (eupatilin, eupatorin, 3'-methyleupatorin, apigenin and isokaempferid), lignans (arctigenin, arctiin and matairesinol), the sesquiterpene cnicin, serotonin conjugates (moschamine and cis-moschamine), β-amyrin and β-sitosterin-β-D-glycopyranoside, identified by means of UV, MS and NMR spectroscopy, were obtained for the first time from this species. The isolated compounds were also evaluated for their tumour cell growth inhibitory activities on HeLa, MCF7 and A431 cells, and different types of secondary metabolites were found to be responsible for the antitumour effects of the extracts; in addition to moderately active compounds (isokaempferid and moschamine), especially apigenin, eupatorin, arctigenin, arctiin, matairesinol and cnicin exert marked antitumour effects against these cell lines.

Abstract  In vitro studies were conducted to investigate the anti-staphylococcal activity of crude extracts of two strains of Pisolithus albus collected from different hosts in the vicinity of Pune University Campus, Pune, India. Organic solvents of increasing polarities were used for extraction of secondary metabolites. In addition, bioactive components of known antimicrobial activity were isolated and tested against thirty strains of clinical isolates of methicillin resistant Staphylococcus aureus. Strong activity was exhibited by ethyl acetate and methanol extracts while aqueous extract showed weak activity. All strains were resistant to the chloroform extract. Maximum effectiveness was recorded for sesquiterpenoids extract (zone diameter: 16.67 to 31.5 mm). Triterpenoids and diterpenoids extracts displayed weak to moderate activity. Polysaccharides fractions IIIa and IIIb were weakly effective while fractions I and II did not show activity. Minimal inhibitory concentration was determined to be in a range of 0.62 to 1.2 mg/ml.

Abstract  Struchium sparganophora (Linn) Ktze, (Asteraceae) is a culinary herbs used as part of a traditional dish in Nigeria and a medicinal plants for the treatment of different ailments in Africa. Dried leaf, stem and root parts of this plant were extracted with n-hexane, chloroform and methanol respectively, concentrated under reduced pressure, freeze dried and evaluated for their antimicrobial and anti tumour activities. The antimicrobial test involved microdilution titre technique while cytotoxicity activities was evaluated using the 3,-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay. The extracts exhibited a broad spectrum of activity against Gram- positive and Gram- negative organism of minimum inhibitory concentration (MIC) of 50 to 6.25 mg/ml but this activity is less than that of anti-microbial standard drug amoxicillin which had MIC of 0.78 to 6.25 mg/ml except against Klebsiella aerogenes on which the extracts had a similar activity to that amoxicillin of MIC 6.25 mg/ml. The extracts showed a significant anti- fungal activity (P< 0.05) of MIC 6.25 mg/ml in comparison with the anti- fungal standard drug Fluconazole (MIC) 50 mg/ml. The extracts also showed to be cytotoxic to all cancer cell lines at the dose of 0.75 mg/ml and significant growth inhibitory activity to all the cancer cell lines used for the test at the dose of 1 mg/ml. The hexane extracts exhibited increase in most cancer cell lines growth at the doses of 0.1 and 0.25 mg/ml while the chloroform and methanol extracts did not show uniform activity. These activities may portend a beneficial effect to human consumption as it may hinder the development and growth of cancerous cells at a high dose.

Abstract  Myristica andamanica is grown only in the Andaman and Nicobar Islands in India. In order to evaluate the wound healing activity of M. andamanica, five different solvent extracts were prepared from the leaves of the plant. Methanol, ethanol, ethyl acetate, chloroform and hexane were used for the extraction of the active ingredients. Excision wound model on Swiss albino mice was used to assess the wound healing activity of the leaves. Remarkable wound healing activity was observed with the ointment formulation of the methanol extract at 1% concentration. Wound contraction was calculated as percentage of the reduction in wound area. A specimen sample of tissue was isolated from the healed skin of each group of mice for the histopathological examination. The results of histopathological examination also supported the outcome of excision wound models. Hematoxylin & Eosin stained sections and Van Gieson's stained sections were checked for collagen deposition. Toluidine blue stained sections were checked for metachromatic staining of mast cells. The present study demonstrated that the aerial parts of M. andamanica promote wound healing activity in mice as a preclinical study.

Abstract  Poly(p-phenylenevinylene) (PPV) derivatives have long been studied because of their attractive opto- and electroluminescent properties and have potential applications for devices such as light-emitting diodes and photovoltaics. The ability to induce alignment of these PPV derivatives may lead to the enhancement of charge mobility and their efficiency. In this study, uniform nanofibers of poly[2,5-(2'-ethylhexyloxy)]-1,4-phenylenevinylene (BEH-PPV) have been fabricated through the method of electrospinning, and an induced alignment of the polymer fibers was observed through photoluminescence data. This study also focuses on the doping of these fibers with the fullerene derivative, 1-(3-methoxycarbonyl)-propyl-1-phenyl-(6,6)-C(61) (PCBM), to induce more incidence of donor/acceptor junctions. Composite fibers with up to a 1:2 weight ratio of PCBM/BEH-PPV have been fabricated and exhibited an ability to quench the photoluminescence of BEH-PPV, indicative of charge transfer.

Abstract  The aim of the present study was to determine the in vivo hypoglycemic activity of five organic extracts and enhydrin obtained from yacon leaves. The main constituents of the most active fraction were identified. Five organic extracts and pure crystalline enhydrin were administered to normoglycemic, transiently hyperglycemic and streptozotocin (STZ)-diabetic rats. The fasting and post-prandial blood glucose, and serum insulin levels were estimated and an oral glucose tolerance test (OGTT) was performed for the evaluation of hypoglycemic activity and dose optimization of each extract. We found that the methanol, butanol and chloroform extracts showed effective hypoglycemic activity at minimum doses of 50, 10 and 20mg/kg body weight, respectively, and were selected for further experiments. Oral administration of a single-dose of each extract produced a slight lowering effect in the fasting blood glucose level of normal healthy rats, whereas each extract tempered significantly the hyperglycemic peak after food ingestion. Daily administration of each extract for 8 weeks produced an effective glycemic control in diabetic animals with an increase in the plasma insulin level. Phytochemical analysis of the most active fraction, the butanol extract, showed that caffeic, chlorogenic and three dicaffeoilquinic acids were significant components. Additionally, enhydrin, the major sesquiterpene lactone of yacon leaves, was also effective to reduce post-prandial glucose and useful in the treatment of diabetic animals (minimum dose: 0.8mg/kg body weight). The results presented here strongly support the notion that the phenolic compounds above as well as enhydrin are important hypoglycemic principles of yacon leaves that could ameliorate the diabetic state.

Abstract  PURPOSE: To understand various soft contact lens materials' ability to adsorb common tear lipids.

METHODS: Ten unworn polymers of nine types were individually soaked in 1.0 mL of 1.75 μg/mL cholesterol oleate or 1.0 mL of 0.5 μg/mL phosphatidylcholine solutions for 1 or 14 days. The adsorbed lipids were extracted with chloroform-methanol, which underwent assay quantification for cholesterol oleate and inorganic phosphate.

RESULTS: More phosphatidylcholine was extracted after 14 days than after 1 day in only lotrafilcon B, balafilcon A, and enfilcon A (all P < 0.005). After 1 day of incubation in phosphatidylcholine, 0.54 to 4.17 μg/lens phosphatidylcholine was recovered from the polymers, and after 14 days of incubation in phosphatidylcholine, 0.58 to 5.77 μg/lens phosphatidylcholine was recovered from the polymers. Etafilcon A had significantly more cholesterol oleate at 1 day than at 14 days, and lotrafilcon A, lotrafilcon B, and balafilcon A had significantly more cholesterol oleate at 14 days than at 1 day (P < 0.005). After 1 day of incubation in cholesterol oleate, 0.14 to 4.80 μg/lens cholesterol oleate was recovered from the polymers. After 14 days of incubation in cholesterol oleate, 1.40 to 6.84 μg/lens cholesterol oleate was recovered from the polymers.

CONCLUSIONS: Hydrogel and most silicone hydrogels appear to adsorb lipids relatively quickly (i.e., within the first day). Although there is some variability in the amounts recovered across materials, it is uncertain whether these differences have any clinical significance.

Abstract  A simple and fast sample preparation method for the determination of nonylphenol (NP) and octylphenol (OP) in aqueous samples by simultaneous derivatization and dispersive liquid-liquid microextraction (DLLME) was investigated using gas chromatography-mass spectrometry (GC/MS). In this method, a combined dispersant/derivatization catalyst (methanol/pyridine mixture) was firstly added to an aqueous sample, following which a derivatization reagent/extraction solvent (methyl chloroformate/chloroform) was rapidly injected to combine in situ derivatization and extraction in a single step. After centrifuging, the sedimented phase containing the analytes was injected into the GC port by autosampler for analysis. Several parameters, such as extraction solvent, dispersant solvent, amount of derivatization reagent, derivatization and extraction time, pH, and ionic strength were optimized to obtain higher sensitivity for the detection of NP and OP. Under the optimized conditions, good linearity was observed in the range of 0.1-1000 μg L⁻¹ and 0.01-100 μg L⁻¹ with the limits of detection (LOD) of 0.03 μg L⁻¹ and 0.002 μg L⁻¹ for NP and OP, respectively. Water samples collected from the Pearl River were analyzed with the proposed method, the concentrations of NP and OP were found to be 2.40 ± 0.16 μg L⁻¹ and 0.037 ± 0.001 μg L⁻¹, respectively. The relative recoveries of the water samples spiked with different concentrations of NP and OP were in the range of 88.3-106.7%. Compared with SPME and SPE, the proposed method can be successfully applied to the rapid and convenient determination of NP and OP in aqueous samples.

Abstract  Formalin-fixed tissues represent the most abundant clinical material for retrospective studies. However, formalin highly affects macromolecules, impairing their extraction and analysis. In this study, the suitability of some potential substitutes of formalin for RNA-based applications has been considered. Conventional formalin was compared with methacarn and the commercial FineFIX. Their impact on overall RNA preservation was investigated in a cell line-based model fixed during a time course treatment and in a series of fixed human tissues. RNA yield was detected by Nanodrop; ribosomal RNA (rRNA) integrity by electrophoresis and the Agilent Bioanalyzer; messenger RNA (mRNA) integrity by Northern blot and endpoint reverse transcription-polymerase chain reaction; and mRNA amount by real-time polymerase chain reaction. In the cell line model, formalin fixation showed time-dependent detrimental effects on overall RNA preservation. Methacarn and FineFIX were more conservative on both rRNA and mRNA preservation and their impact was time-independent. In tissues, high rRNA degradation levels were found in all fixed specimens, contrasting with the results found in the cells. Conversely, the effects of the fixatives on mRNA integrity reflected the observations shown in the cell line model. In methacarn-fixed samples mRNA amount was also preserved, whereas in formalin and FineFIX-fixed samples it was notably lower when compared with the fresh frozen control. Alcohol-based fixatives are a good solution for long-term fixation of both cytologic and tissue samples by virtue of their time-independent effects on mRNA preservation. In fixed tissue samples, however, the potential effects of preanalytical tissue-related factors should be considered when performing mRNA quantitative analysis.

Abstract  Several lines of evidence suggest that detergent-resistant membranes (DRMs) (also known as lipid rafts and glycosphingolipid-enriched microdomains) may have a role in signaling pathways of apoptosis. Here, we developed a method that combines DRMs isolation and methanol/chloroform extraction with stable isotope labeling with amino acids in cell culture-based quantitative proteome analysis of DRMs from control and cisplatin-induced apoptotic Jurkat T cells. This approach enabled us to enrich proteins with a pivotal role in cell signaling of which several were found with increased or decreased amounts in DRMs upon induction of apoptosis. Specifically, we show that three isoforms of protein kinase C (PKC) are regulated differently upon apoptosis. Although PKC alpha which belongs to the group of conventional PKCs is highly up-regulated in DRMs, the levels of two novel PKCs, PKC eta and PKC theta, are significantly reduced. These alterations/differences in PKC regulation are verified by immunoblotting and confocal microscopy. In addition, a specific enrichment of PKC alpha in apoptotic blebs and buds is shown. Furthermore, we observe an increased expression of ecto-PKC alpha as a result of exposure to cisplatin using flow cytometry. Our results demonstrate that in-depth proteomic analysis of DRMs provides a tool to study differential localization and regulation of signaling molecules important in health and disease.

Abstract  Anaemia is a serious health problem especially in developing countries. In the tropics, rural dwellers had often resort to herbal treatments in some cases of anaemia. The essence of this work was to ascertain the rational for the use of the leaves of Brillantaisia nitens as a traditional hematinic. The aqueous extract (AE), methanol extract (ME), n-Hexane fraction (HF), chloroform fraction (CF) and methanol fraction (MF) of the leaves were tested for hematinic effects using phenylhydrazine (PHZ) - induced anaemia in rats. Blood parameters such as red blood cell (RBC) count, white blood cell (WBC) count, hemoglobin concentration (Hb) and haematocrit were assayed as indices of anaemia. Analysis of the vitamin and mineral contents of the ME and MF were carried out while phytochemical analysis of the extracts and fractions as well as the LD(50) of the ME were determined. The results indicated that the oral administration of AE, ME, CF, HF and MF (400 - 3200 mg/kg/day) exhibited a significant (P < 0.05) hematinic activity by ameliorating PHZ - induced decrease in blood parameters viz, Hb, PCV, WBC and RBC. The hematinic potency was in the order of MF > ME > AE > CF > HF. Vitamin analysis showed that both ME and MF contain adequate amounts of water soluble vitamins (Vitamins B(6), B(12), C, and Folic acid), Vit. E and iron in varying concentrations. Phytochemical analysis of the extracts and fractions indicated high concentration of glycosides, alkaloids and resins while flavonoids, terpenoids, carbohydrates and saponins occurred in trace amounts. The oral LD(50) value of the ME was greater than 5000 mg/kg, indicating the high safety profile of B. nitens. These results conclude the presence of hematinic principles in the leaves of B. nitens, thus justifying its use in the management of anemia especially in children.

Abstract  To discover new therapeutically effective and safe cholinesterase and lipoxygenase inhibitors, crude methanol extract and subsequent solvent fractions of Withania somnifera Dunal were screened for their inhibitory profile. Results revealed marked overall inhibition. Against acetylcholinesterase, the IC(50) values were in the range of 69 to 111 mu g/ml, except hexane fraction. Similarly, profound lipoxygenase attenuation was observed (IC(50): 76 to 132 mu g/ml) for extracts with the exception of hexane fraction. It is notable that among the extracts, chloroform fraction was the most dominant inhibitor of the tested enzymes. This study indeed provides a strong scientific rationale for the folk use of the plant for the loss of memory and as anti-inflammatory.

Abstract  In this study, core-shell magnetic mesoporous microspheres with C18-functionalized interior pore-walls were synthesized through coating Fe sub(3)O sub(4) microspheres with a mesoporous inorganic-organic hybrid layer with a n-octadecyltriethoxysilane (C18TES) and tetraethyl orthosilicate (TEOS) as the silica source and cetyltrimethylammonia bromide (CTAB) as a template. The obtained C18-functionalized Fe sub(3)O sub(4)mSiO sub(2) microspheres possess numerous C18 groups anchored in the interior pore-walls, large surface area (274.7 m super(2)/g, high magnetization (40.8 emu/g) and superparamagnetism, uniform mesopores (4.1 nm), which makes them ideal absorbents for simple, fast, and efficient extraction and enrichment of hydrophobic organic compounds in water samples. Several kinds of phthalates were used as the model hydrophobic organic compounds to systematically evaluate the performance of the C18-functionalized Fe sub(3)O sub(4)mSiO sub(2) microspheres in extracting hydrophobic molecules by using a gas chromatography-mass spectrometry. Various parameters, including eluting solvent, the amounts of absorbents, extraction time and elution time were optimized. Hydrophobic extraction was performed in the interior pore of magnetic mesoporous microspheres, and the materials had the anti-interference ability to macromolecular proteins, which was also investigated in the work. Under the optimized conditions, C18-functionalized Fe sub(3)O sub(4)mSiO sub(2) microspheres were successfully used to analyze the real water samples. The results indicated that this novel method was fast, convenient and efficient for the target compounds and could avoid being interfered by macromolecules.

Abstract  A rapid, sensitive, and simple high-performance liquid chromatographic (HPLC) method with an ultraviolet detector (UV) has been developed for the determination of oxaliplatin in the plasma of rabbits and tissues of mice. The sample preparation was carried out by complexation with 0.5 mL of DETC (diethyl-dithiocarbamate) solution and extracted by ether and chloroform. Then, 20 microL of supernatant was injected into the HPLC system with 0.25 mol/L of sodium chloride solution and methanol (30:70 v/v) as the mobile phase at a flow rate of 1.0 mL/min. Separation was performed with a C(18) column at 25 degrees C. The peak was detected at 254 nm. The calibration curve was linear (R(2) > or = 0.9995) in the concentration range of 0.1 approximately 200 microg/mL in plasma and tissues. The intra- and interday variation coefficients were not more than 2.61 and 3.83%, respectively. The limit of detection was 20 ng/mL. The mean recoveries of oxaliplatin were ranged from 97.83 to 104.17% in plasma and tissues. The present method has been successfully applied to the pharmacokinetic study of oxaliplatin liposome in mice and rabbits.

Abstract  The diuretic and laxative activity of different extracts of the barks of Spondias pinnata (Linn. f) Kurz (Family: Rubiaceae) were studied in Wistar albino rats. Furosemide (10 mg/kg, p.o.) and agar-agar (300 mg/kg, p.o.) were used as reference standards respectively for activity comparison. The chloroform and methanol extracts produced significant diuretic and laxative activity. On the other hand, the petroleum ether extract did not reveal significant activity. Urinary levels of sodium, potassium (by flame photometry) and chloride (by titrimetry) were estimated.

Abstract  The methanol and methanol-chloroform (1:1) extracts of the freshly collected Haliclona exigua showed minumim inhibitory concentration (MIC) of 125 mu g/ml and 250 mu g/ml respectively in in vitro studies, but when both of these were tested in vivo in rats, only methanol-chloroform showed 80% inhibition of trophozoites at the dose of 900 mg/kg body weight against Entameba histolytica. Therefore only methanol-chloroform extract was further fractionated into four fractions (n-hexane, chloroform, n-butanol soluble and n-butanol insoluble fractions). Out of these, only n-hexane and n-butanol soluble fractions showed 80% inhibition of trophozoites at 900 mg/kg dose. Further the chromatography of the n-butanol fraction yielded araguspongin-C which showed promising results at different doses.

Abstract  SDS has recognized benefits for protein sample preparation, including solubilization and imparting molecular weight separation (e.g., SDS-PAGE). Here, we compare two proteome workflows which incorporate SDS for protein separation, namely, SDS-PAGE coupled to LC/MS (GeLC MS), along with a solution separation platform, GELFrEE, for intact proteome prefractionation and identification. Despite the clear importance of SDS in these and other proteome analysis workflows, the affect of SDS on an LC/MS proteome experiment has not been quantified. We first examined the influence of SDS on both a bottom-up as well as a top-down (intact protein) MS workflow. Surprisingly, at levels up to 0.01% SDS in the injected sample, reliable MS characterization is obtained. We subsequently explored organic precipitation protocols (chloroform/methanol/water and acetone) as a means of lowering SDS, and present a simple modified acetone precipitation protocol which consistently enables MS proteome characterizations from samples initially containing 2% SDS. With this effective strategy for SDS reduction, the GELFrEE MS workflow for bottom-up proteome analysis was characterized relative to GeLC MS. Remarkable agreement in the number and type of identified proteins was obtained from these two separation platforms, validating the use of SDS in solution-phase proteome analysis.

Abstract  This study was conducted to search for xanthine oxidase (XO) inhibitors from the root extracts of Tamus communis L. traditionally used in folk medicine in Algeria. Root extracts with different solvents were screened for purified milk xanthine oxidase inhibition. The root extracts (methanol, chloroform and ethyl acetate) and proteins, obtained in distilled water, inhibited bovine, sheep and human milk XO from three species in a concentration-dependent manner, with an additional superoxide scavenging capacity, which reached its highest level with ethyl acetate extract (IC(50) = 0.15, 0.04 and 0.09 g/L) for bovine XO, sheep XO and human XO, respectively. The antioxidant potential was confirmed with the non-enzymatic method, total radical-trapping antioxidant parameter (TRAP) assay, which showed that the Tamus communis L. extracts have a potential antioxidant activity in the same order obtained by using the reduction of cytochrome c, an enzymatic method, in which the antioxidant activity followed a decreasing order: ethyl acetate extract > chloroform extract > protein.