Determination of 5-hydroxymethyl-2-furfural,albiflorin, paeoniflorin,liquiritin, ferulic acid,nodakenin, and glycyrrhizin by HPLC-PDA,and evaluation of the cytotoxicity of Palmul-tang,a traditional Korean herbal medicine

A high-performance liquid chromatographic (HPLC) method was developed for quantitative analysis of seven components, 5-hydroxymethyl-2-furaldehyde (1), albiflorin (2), paeoniflorin (3), liquiritin (4), ferulic acid (5), nodakenin (6), and glycyrrhizin (7) of Palmul-tang (PMT), a traditional Korean medicine. HPLC analysis was performed using a Gemini C18 column at 40°C, and photodiode array (PDA) detection at 230 nm, 254 nm, 280 nm, 320 nm, and 330 nm was used for quantification of the seven components in PMT. The mobile phase was a gradient flow composed of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile containing 1.0% (v/v) acetic acid. Calibration curves were acquired with r ² values > 0.9998, and the relative standard deviations (RSDs, %) for intra- and interday precision were both less than 6.0%. The recovery of each component was in the range of 90.66–103.79%, with a RSD less than 5.0%. The contents of the seven components in PMT range form 0.61–6.21 mg/g. Additionally, we investigated the cytotoxicity of the extract against the RBL-1 and BEAS-2B cell lines, as well as splenocytes.     

New C-19-modified geldanamycin derivatives: synthesis,antitumor activities,and physical properties study

Thiazinogeldanamycin (2) was identified from Streptomyces hygroscopicus 17997 at the late stage of the fermentation. The pH was firstly proposed as an important factor in the biosynthesis of it. It was verified that 2 was produced by direct chemical reactions between geldanamycin (1, GDM) and cysteine or aminoethanethiol hydrochloride at pH > 7 in vitro. The proposed synthesis pathway for compound 2 was also discussed. Eleven new C-19-modified GDM derivatives, including five stable hydroquinone form derivatives, were synthesized, most of which exhibited desirable properties such as lower cytotoxicity, increased water solubility, and potent antitumor activity. Especially, compounds 5 and 8 showed antitumor activities against HepG2 cell with IC₅₀ values of 2.97–6.61 μM, lower cytotoxicity and at least 15-fold higher water solubility compared with 1 in pH 7.0 phosphate buffer.     

Design,synthesis, and antitumor activity of oleanolic acid derivatives

Using the techniques of computer-aided drug design, the docking of survivin and known active small molecules was simulated and then the key amino acid residue fragments of the target protein were analyzed. It led to the discovery of active groups capable of binding to the critical sites. Through the use of the natural product, oleanolic acid, as a lead compound, the introduction of the active groups onto the A-ring, and the modification of the carboxyl group at the C-28 position using esterification or amidation, 20 new oleanolic acid derivatives had been designed and synthesized. HepG2 and SGC-7901 cells were used to screen the antitumor activity through the standard MTT method. The compounds, II₃, III₅ and IV_4, exhibited more potent cytotoxicity than positive drugs. Western blot experiment demonstrated that compound II₃ can effectively inhibit the proliferation of HepG2 cells.     

Anti-acne activities of pulsaquinone, hydropulsaquinone, and structurally related 1, 4-quinone derivatives

Quinone type compound, pulsaquinone 1, isolated from the aqueous ethanol extract of the roots of Pulsatilla koreana exhibited antimicrobial activities against an anaerobic non-spore-forming gram-positive bacillus, Propionibacterium acnes, which is related with the pathogenesis of the inflamed lesions in a common skin disease, acne vulgaris. Compound 1 was unstable on standing and thus converted to more stable compound 2, namely hydropulsaquinone by hydrogenation, whose activity was comparable to mother compound 1 (MIC for 1 and 2 against P. acnes: 2.0 and 4.0 μg/mL, respectively). Other structurally-related quinone derivatives (3–13) were also tested for structure-activity relationship against anaerobic and aerobic bacteria, and fungi. The antimicrobial activity was fairly good when the quinone moiety was fused with a nonpolar 6- or 7-membered ring on the right side whether or not conjugated (1,4-naphtoquinone derivatives 3–5), while simple quinone compounds 6–9 showed poor activity. It seems that the methoxy groups at the left side of the quinone function deliver no considerable antimicrobial effect.     

Speciosaperoxide,a new triterpene acid,and other terpenoids from Chaenomeles speciosa

Investigation on the EtOH extract of the fruits of Chaenomeles speciosa led to the isolation of a new triterpene acid bearing an unusual hydroperoxyl substitute group at C-11, speciosaperoxide (1), along with six known triterpenoids, 3β-acetoxyurs-11-en-13β,28-olide (2), 3-O-acetyl ursolic acid (3), oleanolic acid (4), ursolic acid (5), masilinic acid (6), and tormentic acid (7), and three known norsesquiterpenoids, roseoside (8), vomifoliol (9) and (6S,7E,9R)-6,9-dihydroxy-4,7-megastigmadien-3-one 9-O-[β-d -xylopyranosyl (1 → 6)-glucopyranoside] (10). Their structures were elucidated on the basis of spectroscopic data and comparison with reference data. Besides compound 1, compounds 2, 8–10 were obtained from this genus for the first time. None of these compounds exhibited inhibitory activity against T-and B-lymphocyte proliferation.     

Synthesis and antitumor,antityrosinase, and antioxidant activities of xanthone

Ten substituted 1,3-dihydroxyxanthones were synthesized in one step. The yields ranged from 40 to 76%. Compounds 8–10 were first reported. Next, the compounds’ in vitro anti-proliferative activities against nine human cancer cell lines, antityrosinase, and antioxidant activities were evaluated. Compounds 1, 4, 6–7, and 9–10 exhibited enhanced cytotoxicity against certain cancer cells. Compounds 2, 8, 9, and 10 inhibited tyrosinase activity to a certain extent. In addition, compound 4 exhibited the best antioxidant activity, which was consistent with theoretical calculations. These results demonstrated that compounds 1–2, 4, and 6–10 were promising leads for further investigation.     

Analgesic,antipyretic, anti-inflammatory and toxic effects of andrographolide derivatives in experimental animals

Andrographolide (1) and 14-deoxy-11,12-didehydroandrographolide (2) are active constituents of Andrographis paniculata (Burm. f.), family Acanthaceae. A. paniculata extracts are reported to have antiviral, antipyretic, immunostimulant and anticancer activities. In this study, 1 and its 14-acetyl- (4) and 3,19-isopropylidenyl- (3) derivatives, as well as 2 and its 3,19-dipalmitoyl-derivative (5), were intraperitoneally tested for their analgesic, antipyretic, anti-inflammatory and acute toxicity effects in animal models. Analgesic effects were tested in mice using hot plate and writhing tests to distinguish the central and peripheral effects, respectively. The results showed that, at 4 mg/kg, all tested substances have significant analgesic effects, and the highest potency was seen with 3, 4 and 5. Increasing the dose of 3 and 5 to 8 mg/kg did not increase the analgesic effect. In the writhing test, 3 and 5, but not 1, showed significant results. In a baker’s yeast-induced fever model, 3 and 5 significantly reduced rats’ rectal temperature (p < 0.05). In a carrageenan-induced inflammation model, 1, 3 and 5 significantly reduced rats’ paw volume. Doses of 3 and 5 up to 100 mg/kg did not show any serious toxic effects. From this study, 3 and 5 are the most interesting derivatives, showing much greater potency than their parent compounds. These could be further developed as analgesic, antipyretic and anti-inflammatory agents, without any serious toxicity.     

Synthesis, characterization, antibacterial, antifungal, and immunomodulating activities of gatifloxacin derivatives

Gatifloxacin is a synthetic broad-spectrum fluorquinolone antibacterial agent with a 3-methylpiperazinyl-side chain at position 7 and a methoxy group at position 8 of the quinolone ring. In the present study different analogues of gatifloxacin were prepared; the piperazinyl ring was chosen as the center of reaction for synthesizing this series of derivatives. The structures of these derivatives were established using spectroscopic techniques such as IR, ¹H NMR, and EIMS. In vitro antibacterial and antifungal activities were evaluated by disc diffusion method and these derivatives were compared with in-use fluoroquinolones like gatifloxacin, sparfloxacin, and gemifloxacin. Derivative A proved very potent against Gram-negative organisms, especially Pseudomonas aeruginosa, Shigella flexeneri, and Klebseilla pneumoniae, and derivatives A–C exhibited good antifungal activity compared to in-use quinolones. In addition, gatifloxacin and derivatives were investigated for immunomodulating activities. Derivative B has good anti-inflammatory activity, with IC₅₀ < 12.5 μg/ml.     

DNA Damage in T and B Lymphocytes,Bone Marrow,Spleens, and Livers of Rats Exposed to Benzene

Abstract

Single-cell gel electrophoresis assays were performed in order to evaluate DNA damage occurring in the T and B lymphocytes, spleens, bone marrow, and livers of rats exposed to benzene at a concentration of 100, 200, or 400 ppm for 2 or 4 wk. The level of t,t-muconic acid (t,t-MA), which is a urinary benzene metabolite, was determined. In the control rats, mean Olive tail moments in the T and B lymphocytes were 1.507 ± 0.398 and 1.579 ± 0.206, respectively. DNA damage in the T and B lymphocytes exposed to 400 ppm benzene for 4 wk caused those rats to exhibit the highest Olive tail moments, with their values measured as 4.351 ± 0.510 and 3.140 ± 0.631, respectively. Also, the t,t-MA levels increased directly with increasing benzene exposure time and dose during the 4 wk. After 4 wk, the levels of t,t-MA in urine from rats exposed to 100, 200, and 400 ppm were 19.30 ± 5.62, 30.36 ± 4.46, and 46.93 ± 9.10 mg/g creatinine. In conclusion, the present study demonstrates that benzene exposure results in significant DNA damage in the T and B lymphocytes, bone marrow, spleens, and livers of rats. DNA damage in the blood cells and organs was also discovered to vary directly with benzene exposure, in both a dose-dependent and time-dependent manner. In addition, a similar trend regarding DNA damage was found in the blood cells and organs, and evidenced a good association with the level of t,t-MA in the urine.     

Effect of proliferation,cell cycle,and Bcl-2s of MCF-7 cells by resveratrol

Our goals were to examine the dual-directional regulation effects of resveratrol (1) in vitro by using MCF-7 cells (estradiol receptor-positive cells), study its mechanism of action, and give a systematical analysis of the regulatory networks of each related factor. An MTT test and growth curve showed that the proliferation of MCF-7 cells was inhibited by a high concentration of 1, and that its IC₅₀ was 8.70 × 10^? 5 ± 0.23 mol/l. However, 1 induced the proliferation of MCF-7 cells at 10^? 7–10^? 5 mol/l, and resulted in a peak proliferation at 1.0 × 10^? 7 mol/l. A high concentration of 1 arrested cell cycle progression at the G₁ phase, and a typical “sub-G₁ peak” of apoptotic cells was also observed by flow cytometry. The proliferation index of MCF-7 cells increased significantly with a low concentration of 1 (p < 0.05). 1 in high concentrations induced Bax, caspase-3, and cyclin-dependent kinase (CDK) inhibitor P21 expression, whereas the expressions of cyclin CDK2, Bcl-2, and proliferating cell nuclear antigen (PCNA) were decreased by 1 treatment. Conversely, treatment with low concentrations of 1 decreased the expression of P21 and Bax, while the expressions of cyclin CDK2, Bcl-2, and PCNA were increased. These results suggest that 1 had a dual-regulatory effect on MCF-7 cells. CDK-associated protein was a key factor at both the high and low concentrations used in this study.     

Preparation and antitussive,expectorant, and antiasthmatic activities of verticinone's derivatives

To prepare verticinone derivatives with significant antitussive, expectorant, and antiasthmatic activities, the compounds 3β-acetylverticinone (1), 3-ketoverticinone (2), 3β-benzoylverticinone (3), 3β-propionylverticinone (4), 3β-butyrylverticinone (5), and 3β-butoxycarbonylverticinone (6) have been prepared. All of these are new compounds. Among them, 1–6 exhibited potent antitussive and expectorant activities; 1 and 3–6 displayed various antiasthmatic activities. The antitussive activity of 1–6, the expectorant activity of 1–2 and 4–6, and the antiasthmatic activity of 1 are higher than those of verticinone. The results demonstrated that 1 had dominant biological activities, suggesting that it would be a potential antitussive, expectorant, and antiasthmatic agent.     

Compounds from the pods of Astragalus armatus with antioxidant,anticholinesterase, antibacterial and phagocytic activities

Context: The phytochemical study and biological activities of Astragalus armatus Willd. subsp. numidicus (Fabaceae) pods, an endemic shrub of Maghreb, are reported.

Objective: This study isolates the secondary metabolites and determines the bioactivities of Astragalus armatus pods.

Materials and methods: The chloroform, ethyl acetate and n-butanol extracts of hydro-ethanolic extracts were studied. Antioxidant activity was investigated using DPPH and ABTS radical scavenging, CUPRAC and ferrous chelating assays at concentrations ranging from 3 to 200?μg/mL. Anticholinesterase activity was determined against acetylcholinesterase and butyrylcholinesterase enzymes at 50, 100 and 200?μg/mL. Antibacterial activity was performed according to minimum inhibitory concentration (MIC) method. Carbon clearance method in albino mice was used for the phagocytic activity at concentrations 50, 70 and 100?mg/kg body weight. Spectroscopic techniques were used to elucidate the compounds.

Results: Ethyl acetate extract afforded a flavonoid (1) while the n-butanol extract gave four flavonoids (2–5), a cyclitol (6) and a cycloartane-type saponin (7). The ethyl acetate extract exhibited highest antioxidant activity in DPPH (IC₅₀: 67.90?±?0.57?μg/mL), ABTS (IC₅₀: 11.30?±?0.09?μg/mL) and CUPRAC (A_0.50: 50.60?±?0.9?μg/mL) assays. The chloroform extract exhibited the best antibacterial activity against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, each with 80?μg/mL MIC values. The n-butanol extract enhanced phagocytic activity.

Discussion and conclusion: Isorhamnetin (1), isorhamnetin-3-O-α-l-rhamnopyranosyl-(1 → 6)-β-d-galactopyranoside (2), isorhamnetin-3-O-β-d-apiofuranosyl-(1 → 2)-[α-l-rhamnopyranosyl-(1 → 6)]-β-d-galactopyranoside (3), kaempferol-3-O-(2,6-di-O-α-l-rhamnopyranosyl)-β-d-galactopyranoside (4), kaempferol-3-O-(2,6-di-O-α-l-rhamnopyranosyl)-β-d-glucopyranoside (5), pinitol (6) and cyclomacroside D (7) were isolated whereas 1, 2, 6 and 7 are reported for the first time from A. armatus.     

Exploring relationships among pharmacy service use,patronage motives,and patient satisfaction

ObjectivesTo describe and identify significant relationships among pharmacy service use, general and service-specific patient satisfaction, pharmacy patronage motives, and marketing awareness in a service-oriented, independent community pharmacy.DesignCross-sectional study.SettingMidwest United States during May through July 2011.ParticipantsStratified random sample of 500 participants.InterventionSelf-reported questionnaire mailed to participants.Main outcome measuresPatient satisfaction, pharmacy service use, patronage motives, marketing awareness, and demographics.ResultsStudy participants were mostly satisfied with the pharmacy services on global and service-specific measures. Patronage motives of relationships, pharmacy atmosphere, and quality previous experience were associated with increased pharmacy service use at the study pharmacy, while a unique service patronage motivation was associated with decreased pharmacy service use at the study pharmacy. Participants citing pharmacy atmosphere and personnel competency as patronage motives did not use pharmacies other than the study pharmacy more often, whereas participants citing unique services as a patronage motive used pharmacies other than the study pharmacy more often. Direct marketing awareness increased pharmacy service awareness but not use.ConclusionOffering unique services may not be enough to bring in patients loyal to all services provided in a pharmacy. Pharmacists should focus on developing strong relationships with patients and conveying competence when delivering appropriate, quality pharmacy services in a professional pharmacy atmosphere.     

Metabolism of the anti-obesity agent, 4-amino-5-ethyl-3-thiophenecarboxylic acid methyl ester hydrochloride,in rats and dogs

1. In 24 h, male rats excreted in urine 1% of an intra-gastric 100mg/kg dose of 4-amino-5-ethyl-3-[4-¹⁴C]thiophenecarboxylic acid methyl ester hydrochloride (I) as unchanged I and 59% as 4-amino-5-ethyl-3-thiophenecarboxylic acid (II), mostly conjugated.

2. In rats dosed intra-duodenally with I (50mg/kg), little I was found in the systemic circulation (< 2μg/ml) but high concentrations (26 μg/ml) were present at five minutes in portal plasma. At five minutes, II was found at 89 and 93 μg/ml in systemic and portal plasma, respectively. First-pass ester hydrolysis by the duodenum and liver may explain the near absence of I and the high concentrations of II in systemic plasma.

3. Dogs which received 30 mg/kg ¹⁴C-I intra-gastrically, excreted 0.3% I, 30.8 II and 6.8% as 5-ethyl-4-(methylamino)-3-thiophenecarboxylic acid (III), the N-methyl derivative of II.

4. Dogs which received approximately equivalent intra-venous or intra-gastric doses of non-radioactive I and II had high plasma concentrations of II but only small concentrations of I. Plasma concentrations of II after intra-gastric doses of non-radioactive I or II were similar, indicating that both compounds are pharmacokinetically equivalent. I may be a prodrug of II.     

The Crystallite-Gel-Model for Microcrystalline Cellulose in Wet-Granulation,Extrusion, and Spheronization

Purpose. A new model for the wet-extrusion/spheronization process with microcrystalline cellulose (MCC) is proposed. The crystallite-gel-model is able to elucidate the unique role of MCC in this process. Many other experimental results, which cannot be explained by the standard model of granulation, the liquid saturation model, give evidence for the crystallite-gel-model. Methods. Pellets were prepared from different types of MCC. Water content during extrusion, power consumption and aspect ratio were correlated. X-ray diffractograms of MCC powders, extrudates and pellets were taken in order to provide information on changes at the single crystallite level. SEM-photographs and leaching studies gave additional information on changes at the particulate level of MCC. Results. At the level of MCC powder particles, dramatic changes occurred during extrusion/spheronization. In contrast to this no changes could be observed at the level of individual crystallites. Conclusions. During granulation and extrusion MCC-particles are thought to be broken down into smaller particles and possibly ultimate single crystallites in the presence of water. The crystallite-gel-model serves as the framework for a new interpretation of the wet-extrusion/ spheronization process. Apart from the ability to explain experimental data published previously in the literature it can be used to develop new experimental plans for further research. Consequently, the crystallite-gel-model exhibits explanatory as well as predictive power.     

Theory and Computer Programs for Calculating Solution pH,Buffer Formula,and Buffer Capacity for Multiple Component System at a Given Ionic Strength and Temperature

Purpose. A theory and computer programs running on Microsoft^® Excel for Windows for calculation of solution pH, buffer formula, and buffer capacity at a given ionic strength and temperature were developed. The theory does not limit the category of buffer components, the number of buffer components, or the number of ionizations for each buffer component. The usefulness of the programs was examined. Methods. The formulas for 7 single component buffer solutions and 2 multiple component buffer solutions composed of citrate, phosphate, Tris, borate, and glycine were calculated. The solution pH values were measured at 25, 40, 55 and 70°C for comparison with the calculated pH values. Results. Of the 108 predictions made, 96 were of pH values within ± 0.1 pH unit of the measured values, at temperatures ranging from 25°C to 70°C and at ionic strengths ranging from 0.1 M to 0.5 M. Conclusions. These programs will be useful for identifying appropriate buffer solutions at various temperatures and/or ionic strengths.     

Generation,Sampling, and Analysis for Low-Level GB (Sarin) and GF (Cyclosarin) Vapor for Inhalation Toxicology Studies

This study tested and optimized various methodologies to generate, sample, and characterize GB and GF test atmospheres in an inhalation chamber, particularly at low vapor levels. A syringe drive/spray atomization system produced vapor concentrations at a range of 1–50 mg/m³. A saturator cell was used to generate vapor at sub-lethal concentrations ranging from 1 mg/m³ down to low levels approaching the threshold limit value time-weighted average (TLV-TWA) of 0.0001 mg/m³ for GB. Both generation techniques demonstrated the ability to produce stable vapor concentrations over extended exposure periods. This capability was important to determine sublethal nerve agent effects, such as miosis, for inhalation toxicology studies. In addition, the techniques employed for producing and maintaining low-level agent vapor would lay the foundation for testing less volatile chemical warfare agents such as VX.     

Alcohol Use,Drug Use,and Sexual Activity among Pharmacy Students at Three Institutions

ObjectiveTo determine alcohol and drug use and sexual activity among pharmacy students at three colleges of pharmacy.Design and SettingA survey to obtain self-reported information on alcohol and drug use, and sexual activity was administered to professional pharmacy students at the University of Iowa (UI), Massachusetts College of Pharmacy, and Texas Southern University.Main Outcome MeasuresInformation on sexual activity and condom use, alcohol and drug consumption, and the effect of alcohol on unintended sexual activity.Results848 students (50% response rate) completed the survey. Alcohol use was high at all three institutions, and most students had consumed five or more drinks on one or more occasions within the last three months. The extent of drug use among pharmacy students was similar to that reported in other college students. The majority of students were sexually active. More men than women reported having been sexually active with one or more partners. Most students reported having had sexual intercourse without a condom. Significant numbers of students had engaged in unintended sex after alcohol use, especially at UI (χ² = 12.6, p = 0.002). Sexual contact and drinking were strongly correlated (Pearson r = 0.31, p = 0.0001).ConclusionAlcohol consumption among pharmacy students was high. Heavy drinking (five or more drinks on one occasion) was associated with unintended sexual contact. Given low condom use and increased sexual contact, pharmacy students are at an increased risk for HIV infection. Strategies should be developed to reduce alcohol intake and unprotected sexual activity among pharmacy students.     

The relationship among depressive symptoms,urgency, and problematic alcohol and cannabis use in community adults

BackgroundThis study examined alternative models for how negative and positive urgency influence the relationship between depressive symptoms and alcohol and cannabis use in a community sample.MethodsParticipants included adults (n = 675; age = 42.57, SD = 15.66; 65.0% female; 74.2% White) in the community sample from the Rockland Project by the Nathan Kline Institute. Path analyses were conducted.ResultsNegative urgency was a unique mediator in the relationship between depressive symptoms and problematic alcohol use (β =0.121, 95% CI = 0.060–0.182) and problematic cannabis use (β =0.120, 95% CI = 0.060–0.179). Negative and positive urgency significantly moderated the relationship between depressive symptoms and problematic cannabis use (negative urgency: β = 0.092, 95% CI = 0.040–0.145; positive urgency: β = 0.070, 95% CI = 0.022–0.119), such that the relationship was positive at high levels and negative at low levels of urgency. The patterns and levels of the relationships between depressive symptoms and cannabis use differed between negative and positive urgency. Neither urgency trait moderated the relationship between depressive symptoms and problematic alcohol use.ConclusionsDespite being strongly related, negative and positive urgency have distinct roles in the relationship between depressive symptoms and problematic alcohol and cannabis use. Previous finding with younger samples that do not include both traits in the model at time generalize and at other times do not replicate, which warrants the continued examination of how these traits impart risk across the lifespan.