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Sample records for vitro schistosomicidal activity

  1. Schistosoma mansoni: in vitro schistosomicidal activity and tegumental alterations induced by piplartine on schistosomula.

    PubMed

    de Moraes, Josué; Nascimento, Carlos; Yamaguchi, Lydia F; Kato, Massuo J; Nakano, Eliana

    2012-10-01

    Schistosomiasis is one of the most important parasitic infections in humans that occur in many tropical and subtropical countries. Currently, the control of schistosomiasis rests with a single drug, praziquantel, which is effective against adult worms but not the larval stages. Recent studies have shown that piplartine, an amide isolated from plants of the genus Piper (Piperaceae), reveals interesting antischistosomal properties against Schistosoma mansoni adult worms. Here, we report the in vitro antischistosomal activity of piplartine on S. mansoni schistosomula of different ages (3 h old and 1, 3, 5, and 7 days old), and examine alterations on the tegumental surface of worms by means of confocal laser scanning microscopy. Piplartine at a concentration of 7.5 ?M caused the death of all schistosomula within 120 h. The lethal effect occurred in a dose-dependent manner and was also dependent on the age of the parasite. Microscopy observation revealed extensive tegumental destruction, including blebbing, granularity, and a shorter body length. This report provides the first evidence that piplartine is able to kill schistosomula of different ages and reinforce that piplartine is a promising compound that could be used for the development of new schistosomicidal agent. PMID:22796749

  2. Schistosomicidal activity and docking of Schistosoma mansoni ATPDase 1 with licoflavone B isolated from Glycyrrhiza inflata (Fabaceae).

    PubMed

    Aleixo de Carvalho, Lara Soares; Geraldo, Reinaldo Barros; de Moraes, Josué; Silva Pinto, Pedro Luiz; de Faria Pinto, Priscila; Pereira, Olavo Dos Santos; Da Silva Filho, Ademar A

    2015-12-01

    Schistosomiasis is one of the world's major public health problems, and its treatment is widely dependent on praziquantel (PZQ), the only available drug. Schistosoma mansoni ATP diphosphohydrolases are ecto-enzymes localized on the external tegumental surface of S. mansoni and considered an important target for action of new drugs. In this work, the in vitro schistosomicidal activity of the crude extract of Glycyrrhiza inflata roots (GI) and its isolated compounds echinatin, licoflavone A and licoflavone B were evaluated against S. mansoni adult worms. Results showed that GI (200 ?g/mL) was active against adult schistosomes, causing 100% mortality after 24 h of incubation. Chromatographic fractionation of GI led to isolation of echinatin, licoflavone A and licoflavone B. Licoflavone B (25-100 ?M) caused 100% mortality, tegumental alterations, and reduction of oviposition and motor activity of all adult worms, without affecting mammalian Vero cells. Confocal laser scanning microscopy showed tegumental morphological alterations and changes on the numbers of tubercles of S. mansoni worms in a dose-dependent manner after incubation with licoflavone B. Licoflavone B also showed high S. mansoni ATPase (IC50 of 23.78 ?M) and ADPase (IC50 of 31.50 ?M) inhibitory activities. Docking studies predicted different interactions between licoflavone B and S. mansoni ATPDase 1, corroborating with the in vitro inhibitory activity. This report demonstrated the first evidence for the schistosomicidal activity of licoflavone B and suggests that its mechanism of action involve the inhibition of S. mansoni ATP diphosphohydrolases. PMID:26454044

  3. Potent Schistosomicidal Constituents from Garcinia brasiliensis.

    PubMed

    Castro, Aline Pereira; de Mattos, Ana Carolina Alves; Pereira, Neusa Araújo; Anchieta, Naira Ferreira; Silva, Matheus Siqueira; Dias, Danielle Ferreira; Silva, Claudinei Alves; Barros, Giulliano Vilela; Souza, Raquel Lopes Martins; Dos Santos, Marcelo Henrique; Marques, Marcos José

    2015-06-01

    Praziquantel is the drug of choice for the treatment of schistosomiasis. However, several strains of Schistosoma mansoni are resistant to praziquantel, making it necessary to discover new drugs that might be used for its treatment. With this in mind, the properties of a schistosomicidal ethanolic extract of Garcinia brasiliensis Mart. epicarp, the fractions obtained by partitioning this extract, including the hexane fractions, ethyl acetate fraction, and the aqueous fraction, and the isolated compounds 7-epiclusianone, a major component from these fractions, and fukugetin were tested in vitro on adult worms of S. mansoni. Mortality, damage to membranes, and excretory system activity were observed at 100.0, 50.0, 75.0, and 14.0?µg/mL for the ethanolic extract of G. brasiliensis Mart. epicarp, its hexane fractions, the ethyl acetate fraction, and 7-epiclusianone, respectively. For 7-epiclusianone, these data were confirmed by fluorescent probe Hoechst 33?258 and resorufin. Additionally, the biocidal effect of 7-epiclusianone was even higher than the hexane fractions. Moreover, an inhibitory effect of 7-epiclusianone on the egg laying of female adult S. mansoni worms was observed in cercariae and schistossomula. Thus, 7-epiclusianone is a promising schistosomicidal compound; however, more studies are needed to elucidate its mechanism of toxicity and to evaluate the in vivo activity of this compound. PMID:25905590

  4. In Vitro and In Vivo Anti-Schistosomal Activity of the Alkylphospholipid Analog Edelfosine

    PubMed Central

    Yepes, Edward; Varela-M, Rubén E.; López-Abán, Julio; Dakir, E. L. Habib; Mollinedo, Faustino; Muro, Antonio

    2014-01-01

    Background Schistosomiasis is a parasitic disease caused by trematodes of the genus Schistosoma. Five species of Schistosoma are known to infect humans, out of which S. haematobium is the most prevalent, causing the chronic parasitic disease schistosomiasis that still represents a major problem of public health in many regions of the world and especially in tropical areas, leading to serious manifestations and mortality in developing countries. Since the 1970s, praziquantel (PZQ) is the drug of choice for the treatment of schistosomiasis, but concerns about relying on a single drug to treat millions of people, and the potential appearance of drug resistance, make identification of alternative schistosomiasis chemotherapies a high priority. Alkylphospholipid analogs (APLs), together with their prototypic molecule edelfosine (EDLF), are a family of synthetic antineoplastic compounds that show additional pharmacological actions, including antiparasitic activities against several protozoan parasites. Methodology/Principal Findings We found APLs ranked edelfosine> perifosine> erucylphosphocholine> miltefosine for their in vitro schistosomicidal activity against adult S. mansoni worms. Edelfosine accumulated mainly in the worm tegument, and led to tegumental alterations, membrane permeabilization, motility impairment, blockade of male-female pairing as well as induction of apoptosis-like processes in cells in the close vicinity to the tegument. Edelfosine oral treatment also showed in vivo schistosomicidal activity and decreased significantly the egg burden in the liver, a key event in schistosomiasis. Conclusions/Significance Our data show that edelfosine is the most potent APL in killing S. mansoni adult worms in vitro. Edelfosine schistosomicidal activity seems to depend on its action on the tegumental structure, leading to tegumental damage, membrane permeabilization and apoptosis-like cell death. Oral administration of edelfosine diminished worm and egg burdens in S. mansoni-infected CD1 mice. Here we report that edelfosine showed promising antischistosomal properties in vitro and in vivo. PMID:25302497

  5. Structurally modified natural sesquiterpene lactones constitute effective and less toxic schistosomicidal compounds.

    PubMed

    Sass, Daiane Cristina; Morais, Gustavo Oliveira; Miranda, Ricardo Augusto Crema; Magalhăes, Lizandra Guidi; Cunha, Wilson Roberto; dos Santos, Raquel Alves; Arakawa, Nilton Syogo; Da Costa, Fernando Batista; Constantino, Mauricio Gomes; Heleno, Vladimir Constantino Gomes

    2014-10-28

    Sesquiterpene lactones are known to be active, but are also known to present high cytotoxicity. In the present work an evaluation of how slight structural alterations affect the cytotoxicity and the schistosomicidal activity of sesquiterpene lactones was undertaken. More specifically, we assessed the activity of budlein-A, a furanoheliangolide sesquiterpene lactone, and four of its derivatives. The structural modifications of budlein-A, presented in this work, diminished the cytotoxicity and changed the antiparasitary behavior of the molecule. They also provided data for a better understanding of the sesquiterpene lactone cytotoxicity. The establishment of the structures of three synthesized sesquiterpene lactones on the basis of NMR and HRESIMS data is also presented here. Complete and detailed (1)H and (13)C 1D and 2D NMR data, with measurements of all J values and all multiplicities clarified, are presented for five sesquiterpene lactones for the first time. PMID:25030079

  6. A study of the distribution of schistosomicidal drug H-3-7505 in mice

    SciTech Connect

    Hao, T.

    1985-05-01

    The authors have studied the distribution of H-3 labelled schistosomicidal drug in mice by autoradiography. The H-3-labelled substances were found in liver and kidney and in successfully decreasing amounts in brain, lung, heart, fat, testis, pancreas and spleen. In various cells the silver granules were present mainly in the cytoplasms but a few in the nucleus. After administration of this labelled schistosomicidal drug, the mice were killed and studied in groups successively at 4, 8, 24 hrs. No difference in the distribution of silver granules were observed. This fact indicated that, this drug was rapidly absorbed and highly concentrated with a long duration of reservation in liver. All of these favours the schistosomicidal effect of the drug. As this drug was highly concentrated in the cytoplasm of liver cells, that might provide a pathophysiologic basis for the explanation of jaundice in the clinical practice. Moreover, the appearance of toxic reaction in nervous system may be related to the relatively high concentration of the drug distributed in the brain.

  7. Exfoliated Egyptian kaolin immobilized heteropolyoxotungstate nanocomposite as an innovative antischistosomal agent: In vivo and in vitro bioactive studies.

    PubMed

    Bayaumy, Fatma E A; Darwish, Atef S

    2016-02-01

    This study aims to manipulate an antischistosomal nanocomposite based on exfoliated clay immobilized heteropolyoxotungstate. The nanocomposite's physicochemical characteristics were examined using XRD, Raman spectroscopy, FTIR, DLS, SEM, HR-TEM and AFM. Nano-sized spheroidal negatively charged Keggin-type heteropolyoxotungstate particles were developed along and between the exfoliated clay layers. The impact of the nanocomposite on Schistosoma mansoni-infected mice was studied through parasitological, physiological and histological analyses. Infected mice were orally vaccinated by a single nanocomposite dose (15mg/kg/day) for two weeks. The schistosomicidal activities of the nanocomposite in vitro were investigated by examining its dose- and time-dependent responses in terms of % worm mortality. The time-dependent morphological alterations in schistosomes at a nanocomposite dosage of 15?g/mL were followed by SEM. The nanocomposite exhibited potential schistosomicidal properties with a marked reduction in worm burden (~85% mortality), extensive deformities in the adult worm tegument and suckers, improvement of serum biochemical activities, and diminishment in granulomatous lesions. The in vitro release of heteropolyoxotungstate from exfoliated clay indicates the clay's ability to embrace the heteropolytungstate until its liberation at the parasitic districts. PMID:26652426

  8. In vitro activity of florphenicol.

    PubMed

    Graham, R; Palmer, D; Pratt, B C; Hart, C A

    1988-10-01

    Florphenicol was active at a lower concentration than chloramphenicol against over half of 234 recent clinical bacterial isolates. The majority (98%) of the isolates were inhibited by florphenicol at a concentration of 8 mg/l or less. Florphenicol was particularly effective against chloramphenicol resistant strains of Haemophilus influenzae. Klebsiella aerogenes and Bacteroides spp. Florphenicol was bacteristatic for salmonellae and Escherichia coli but bactericidal for Haemophilus influenzae. Florphenicol was slightly more active than chloramphenicol against Chlamydia trachomatis, Mycoplasma hominis and Mycoplasma pneumoniae but less active against Ureaplasma urealyticum. PMID:3143587

  9. Analysis of Smad Phosphatase Activity In Vitro.

    PubMed

    Shen, Tao; Qin, Lan; Lin, Xia

    2016-01-01

    Phosphorylation of Smad1/5/8 at the C-terminal SXS motif by BMP type I receptors is one of the most critical events in BMP signaling. Conversely, protein phosphatases that dephosphorylate phospho-Smad1/5/8 can consequently prevent or terminate BMP signaling. PPM1H is an undercharacterized phosphatase in the PPM family. We recently demonstrated that PPM1H can dephosphorylate Smad1 in the cytoplasm and block BMP signaling responses in cellular assays. Here we describe in vitro method showing that PPM1H is a bona fide phosphatase for Smad1/5/8. PPM1H is produced as GST fusion protein in E. coli, and purified against glutathione sepharose beads. Bacterially purified recombinant PPM1H possesses phosphatase activity toward artificial substrate para-nitrophenyl phosphate (pNPP). Recombinant PPM1H also dephosphorylates immuno-purified phosphorylated Smad1 in test tubes. These direct in vitro phosphatase assays provide convincing evidence demonstrating the role of PPM1H as a specific phosphatase for P-Smad1. PMID:26520120

  10. Antiseptic mouthwashes: in vitro antibacterial activity.

    PubMed

    Watanabe, Evandro; Nascimento, Andresa P; Guerreiro-Tanomaru, Juliane M; Razaboni, Ana M; de Andrade, Denise; Tanomaru-Filho, Mário

    2015-08-01

    Mouthwashes are used as an adjunct to tooth brushing for improving breath and preventing oral diseases. The aim of this study was to compare the in vitro Maximum Inhibitory Dilution (MID) of 3 mouthwashes with different active ingredients against mutans streptococci (MS). The products analyzed were PeriogardR, CepacolR and PlaxR Fresh Mint. Their antibacterial activity was assessed in duplicate in 96-well microtiter plates against 36 clinical isolates of MS. Each mouthwash was submitted to a serial two-fold dilution (1/2.5 to 1/5120) using double concentration of Tryptose Soy Broth with 1.0% yeast extract. The final volume in each well was 100 mL plus 5 mL of a bacterial suspension, equivalent to 107 CFU/mL. They were incubated microaerobically at 37oC for 48 hours and the MIDs determined. MID was 1/320 for PeriogardR and CepacolR, and 1/20 for PlaxR. Statistical analysis revealed that the MID of PeriogardR MID did not differ from that of CepacolR (p>0.05), and was higher than that of PlaxR (p<0.05). In conclusion, the antiseptic mouthwashes containing chlorhexidine (PeriogardR) and cetylpyridinium chloride (CepacolR) had higher in vitroantibacterial activity (MID) against MS than the antiseptic mouthwash containing triclosan (PlaxR), according to microbiological method employed. PMID:26355890

  11. HIFU-induced gene activation in vitro

    NASA Astrophysics Data System (ADS)

    Liu, Yunbo; Zhong, Pei; Kon, Takashi; Li, Chuanyuan

    2001-05-01

    This work investigated the inducible gene activation in cancer cells that were sublethally injured during HIFU treatment. HeLa cells were transfected by an adenovirus vector that encodes GFP under the control of hsp70B promoter, leading to about 65% transfection efficiency. A volume of 10 ?L transfected HeLa cells in suspension (5×107 cells/ml) were placed at the bottom of a PCR tube so that the cell suspension could be heated to a peak temperature of 50°C, 60°C, and 70°C for 120, 10, and 1 s, respectively, by a focused 1.1-MHz HIFU transducer operated at a peak negative pressure of -2.7 MPa at different duty cycles. One day after HIFU treatment, cell viability was determined to be 63%, 35%, and 18%, respectively, based on Trypan Blue exclusion test. Importantly, in all test groups, inducible GFP expression was detected in about 40%-50% of the surviving cells with GFP intensity increased by 25-fold based on flow cytometry analysis. These results demonstrate that even under the short exposure duration of HIFU treatment, inducible gene expression could be produced in sublethally injured cell population in vitro. Further studies are underway to explore the optimal HIFU condition for gene activation in vivo.

  12. Suppressive activity of epinastine hydrochloride on eosinophil activation in vitro.

    PubMed

    Mochizuki, Yu-ichiro; Furuta, Atsuko; Furuya, Ayako; Kanai, Ken-ichi; Asano, Kazuhito; Suzaki, Harumi

    2008-01-01

    The influence of a histamine H1 receptor antagonist, epinastine hydrochloride (EP), on eosinophil functions was examined in vitro and in vivo. The first set of experiments was undertaken to examine whether EP could suppress eosinophilia and IgE hyperproduction induced by Mesocestoides cortii infection in BALB/c mice. The number of peripheral blood eosinophils and levels of IgE were examined 21 days after infection. Oral administration of EP at a daily dose of 0.3 mg/kg, which is the recommended human therapeutic dose, for 21 days was not able to suppress either peripheral blood eosinophilia or IgE hyperproduction, which was observed in mice infected with M. cortii. The second part of the experiment was designed to examine the influence of EP on eosinophil activation induced by stem cell factor (SCF) stimulation in vitro. Eosinophils were obtained from M. cortii-infected mice and stimulated with SCF in the presence of different concentrations of EP for 24 h. The addition of EP into cell cultures suppressed eosinophil activation induced by SCF stimulation as assessed by measuring the contents of acronym for Regulated upon Activation, Normal T cell Expressed and presumably Secreted (RANTES), macrophage inflammatory protein-1beta (MIP-1beta) and leukotriene C4 (LTC4) levels in culture supernatants. The minimum concentration of EP which caused significant suppression of factor productions was 25 ng/ml, which is similar to the concentration in plasma after oral administration of the therapeutic dose in humans. These results may suggest that EP exerts inhibitory effects on eosinophil activation and results in favorable modification of the clinical status of allergic patients. PMID:18396776

  13. Evaluation of the Anti-Schistosoma mansoni Activity of Thiosemicarbazones and Thiazoles

    PubMed Central

    de Oliveira, Sheilla Andrade; de Oliveira Filho, Gevânio Bezerra; Moreira, Diogo Rodrigo Magalhaes; Gomes, Paulo André Teixeira; da Silva, Anekécia Lauro; de Barros, Andréia Ferreira; da Silva, Aline Caroline; dos Santos, Thiago André Ramos; Pereira, Valéria Ręgo Alves; Gonçalves, Gabriel Gazzoni Araújo; Brayner, Fábio André; Alves, Luiz Carlos; Wanderley, Almir Gonçalves; Leite, Ana Cristina Lima

    2014-01-01

    Schistosomiasis is a chronic and debilitating disease caused by a trematode of the genus Schistosoma and affects over 207 million people. Chemotherapy is the only immediate recourse for minimizing the prevalence of this disease and involves predominately the administration of a single drug, praziquantel (PZQ). Although PZQ has proven efficacy, there is a recognized need to develop new drugs as schistosomicides since studies have shown that repeated use of this drug in areas of endemicity may cause a temporary reduction in susceptibility in isolates of Schistosoma mansoni. Hydrazones, thiosemicarbazones, phthalimides, and thiazoles are thus regarded as privileged structures used for a broad spectrum of activities and are potential candidates for sources of new drug prototypes. The present study determined the in vitro schistosomicidal activity of 10 molecules containing these structures. During the assays, parameters such motility and mortality, oviposition, morphological changes in the tegument, cytotoxicity, and immunomodulatory activity caused by these compounds were evaluated. The results showed that compounds formed of thiazole and phthalimide led to higher mortality of worms, with a significant decline in motility, inhibition of pairing and oviposition, and a mortality rate of 100% starting from 144 h of exposure. These compounds also stimulated the production of nitric oxide and tumor necrosis factor alpha (TNF-?), thereby demonstrating the presence of immunomodulatory activity. The phthalyl thiazole LpQM-45 caused significant ultrastructural alterations, with destruction of the tegument in both male and female worms. According to the present study, phthalyl thiazole compounds possess antischistosomal activities and should form the basis for future experimental and clinical trials. PMID:24165185

  14. Temporal changes in intestinal Na+ -ATPase activity and in vitro

    E-print Network

    Young, Graham

    Temporal changes in intestinal Na+ , K+ -ATPase activity and in vitro responsiveness to cortisol+ , K+ -ATPase activity were measured in pyloric ceca and posterior intestine of juvenile chinook salmon pronounced increases in endogenous Na+ , K+ -ATPase activity in summer for both intestinal regions

  15. ASSESSMENT OF IN VITRO ANDROGENIC ACTIVITY IN KRAFT MILL EFFLUENT

    EPA Science Inventory

    Detection of In Vitro Androgenic Activity in Feedlot Effluent. Lambright, CS 1 , Guillette, LJ, Jr.2, Gray, LE, Jr.1 , 1USEPA, NHEERL, RTP, NC, 2 University of Florida, Dept. of Zoology, Gainesville FL

    Recent studies have shown the presence of androgenic activity in water...

  16. In vitro antimicrobial activity against 10 North American and European Lawsonia intracellularis isolates

    E-print Network

    Singer, Randall

    In vitro antimicrobial activity against 10 North American and European Lawsonia intracellularis to evaluate the antimicrobial activities of most intracellular organisms since these bacteria only propagate themselves inside the host cell. Therefore, most in vitro studies of antimicrobial activities against

  17. IN VITRO IRON BIOAVAILABILITY AND ANTIOXIDANT ACTIVITY OF RAISINS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Iron bioavailabilities and antioxidant activities of three common generic raisin types, Golden Thompson, Dipped Thompson and Sun dried Thompson, were quantified and compared. Iron bioavailability was assessed with an in vitro digestion/Caco 2 cell culture model using cell ferritin formation as an i...

  18. In vitro anticancer activity of Anemopsis californica.

    PubMed

    Kaminski, Catherine N; Ferrey, Seth L; Lowrey, Timothy; Guerra, Leo; VAN Slambrouck, Severine; Steelant, Wim F A

    2010-07-01

    Three different extract conditions (aqueous, EtOH and EtOAc) of four different parts (bracts, leaves, roots and stems) of the plant Anemopsis californica (A. californica) were evaluated for their effect on the growth and migration of human colon cancer cells, HCT-8, and the breast cancer cell lines Hs 578T and MCF-7/AZ. Our aim was to identify potential anticancer activity in crude A. californica extracts, given that this plant is used by Native Americans to treat a variety of diseases, including cancer. Our results demonstrated that for each of the cell lines tested, the majority of ethyl acetate extracts of all the plant parts are more toxic than the aqueous and ethanol extracts. Furthermore, significant growth inhibitory activity against the three cell lines was found for the ethyl acetate extract of the roots, while the aqueous extract of the roots influenced the migratory capacity of the three cell lines. This study provides evidence for the anticancer properties of A. californica when extracted in water and ethyl acetate, and supports the importance for further purification of the crude extracts and isolation of potential new anticancer compounds through bio-guided fractionation. PMID:21941602

  19. In vitro antioxidant activity of extracts from common legumes.

    PubMed

    Zhao, Yan; Du, Shuang-kui; Wang, Hanxin; Cai, Meng

    2014-01-01

    The in vitro antioxidant activity of pinto bean, cowpea, baby lima bean, lentil, chickpea, small red bean, red kidney bean, black kidney bean, navy bean, and mung bean extracts were investigated. Significant differences were observed in the phenolic content and the antioxidant activity amongst the legume extracts. Lentils demonstrated the highest phenolic content (47.6 mg/g), total antioxidant activity (720.68 U/g), DPPH• scavenging activity (38.5%), and total reducing power, whereas baby lima beans and navy beans had the lowest. Amongst the extracts, hydroxyl radicals (•OH) scavenging was higher in black kidney bean (85.68%) and baby lima bean (74.97%) extracts. The total antioxidant activity (r=0.84), DPPH• scavenging activity (r=0.83), and total reducing power (r=0.84) were positively correlated with the total phenolic content. However, •OH scavenging and the phenolic content were not correlated. PMID:24444962

  20. In vitro modulation of estrogen receptor activity by norfluoxetine

    PubMed Central

    LUPU, DIANA; POP, ANCA; CHERFAN, JULIEN; KISS, BÉLA; LOGHIN, FELICIA

    2015-01-01

    Background and aims Selective serotonin reuptake inhibitors (SSRIs) are antidepressants increasingly prescribed for pregnancy and postpartum depression. However, these compounds can cross the placenta and also pass into breast milk, thus reaching the fetus and infant during critical developmental stages, potentially causing adverse effects. Fluoxetine, a widely used SSRI, has been shown to affect (neuro)endocrine signaling in various organisms, including humans. This compound can also interact with estrogen receptors in vitro and cause an estrogen-dependent uterotrophic response in rodents. Consequently, the aim of the present study was to assess if the active metabolite of fluoxetine, namely norfluoxetine (NFLX), shares the same capacity for estrogen receptor interaction. Methods The in vitro (anti)estrogenic activity of norfluoxetine was assessed using a firefly luciferase reporter construct in the T47D-Kbluc breast cancer cell line. These cells express nuclear estrogen receptors (ERs) that can activate the transcription of the luciferase reporter gene upon binding of ER agonists. Light emission was monitored in case of cells exposed to norfluoxetine or mixtures of norfluoxetine-estradiol. Cell viability was assessed using a resazurin-based assay. Results During individual testing, NFLX was able to induce a significant increase in luciferase activity compared to control, but only at the highest concentration tested (10 ?M). In binary mixtures with estradiol (30 pM constant concentration) a significant increase in luminescence was observed at low submicromolar norfluoxetine concentrations compared to estradiol alone. Conclusion Norfluoxetine can induce estrogenic effects in vitro and can potentiate the activity of estradiol. However, further studies are needed to clarify if these observed estrogenic effects may have detrimental consequences for human exposure. PMID:26609274

  1. Anti-Mycobacterium avium activity of quinolones: in vitro activities.

    PubMed Central

    Klopman, G; Wang, S; Jacobs, M R; Bajaksouzian, S; Edmonds, K; Ellner, J J

    1993-01-01

    The MICs of 88 quinolones against 14 selected reference and clinical strains of Mycobacterium avium-M. intracellular complex were determined. Agents tested included ciprofloxacin, sparfloxacin (PD 131501), and 86 other experimental quinolones. Test strains were selected to represent various susceptibilities to ciprofloxacin and other drug resistance profiles. MICs were determined by the microdilution method in 7HSF broth, with incubation for 14 days at 35 degrees C. The results showed 25 of the quinolones to be active against the strains, with MICs for 90% of the strains (MIC90s) of 2 to 32 micrograms/ml. Ten of these compounds had activities equivalent to or greater than that of ciprofloxacin. The most active compound was PD 125354, with an MIC50 of 0.5 micrograms/ml and an MIC90 of 2 micrograms/ml; comparable values for ciprofloxacin were 4 and 8 micrograms/ml, respectively. The next most active compounds, with MIC90s of 4 micrograms/ml, were sparfloxacin (PD 131501), PD 123982, PD 135144, and PD 119421. MIC90s of PD 131575, PD 126889, PD 122642, PD 139586, and PD 143289 were 8 micrograms/ml. Further evaluation of the most active agents is warranted, as is assessment of structure-activity relationships of active and inactive agents to elucidate the active portions of the compounds and to lead to the development of compounds with enhanced activity. PMID:8239587

  2. Reviews on Mechanisms of In Vitro Antioxidant Activity of Polysaccharides.

    PubMed

    Wang, Junqiao; Hu, Shuzhen; Nie, Shaoping; Yu, Qiang; Xie, Mingyong

    2016-01-01

    It is widely acknowledged that the excessive reactive oxygen species (ROS) or reactive nitrogen species (RNS) induced oxidative stress will cause significant damage to cell structure and biomolecular function, directly or indirectly leading to a number of diseases. The overproduction of ROS/RNS will be balanced by nonenzymatic antioxidants and antioxidant enzymes. Polysaccharide or glycoconjugates derived from natural products are of considerable interest from the viewpoint of potent in vivo and in vitro antioxidant activities recently. Particularly, with regard to the in vitro antioxidant systems, polysaccharides are considered as effective free radical scavenger, reducing agent, and ferrous chelator in most of the reports. However, the underlying mechanisms of these antioxidant actions have not been illustrated systematically and sometimes controversial results appeared among various literatures. To address this issue, we summarized the latest discoveries and advancements in the study of antioxidative polysaccharides and gave a detailed description of the possible mechanisms. PMID:26682009

  3. Reviews on Mechanisms of In Vitro Antioxidant Activity of Polysaccharides

    PubMed Central

    Wang, Junqiao; Hu, Shuzhen; Nie, Shaoping; Yu, Qiang; Xie, Mingyong

    2016-01-01

    It is widely acknowledged that the excessive reactive oxygen species (ROS) or reactive nitrogen species (RNS) induced oxidative stress will cause significant damage to cell structure and biomolecular function, directly or indirectly leading to a number of diseases. The overproduction of ROS/RNS will be balanced by nonenzymatic antioxidants and antioxidant enzymes. Polysaccharide or glycoconjugates derived from natural products are of considerable interest from the viewpoint of potent in vivo and in vitro antioxidant activities recently. Particularly, with regard to the in vitro antioxidant systems, polysaccharides are considered as effective free radical scavenger, reducing agent, and ferrous chelator in most of the reports. However, the underlying mechanisms of these antioxidant actions have not been illustrated systematically and sometimes controversial results appeared among various literatures. To address this issue, we summarized the latest discoveries and advancements in the study of antioxidative polysaccharides and gave a detailed description of the possible mechanisms. PMID:26682009

  4. Amyloid beta modulation of neuronal network activity in vitro.

    PubMed

    Charkhkar, Hamid; Meyyappan, Susheela; Matveeva, Evgenia; Moll, Jonathan R; McHail, Daniel G; Peixoto, Nathalia; Cliff, Richard O; Pancrazio, Joseph J

    2015-12-10

    In vitro assays offer a means of screening potential therapeutics and accelerating the drug development process. Here, we utilized neuronal cultures on planar microelectrode arrays (MEA) as a functional assay to assess the neurotoxicity of amyloid-? 1-42 (A?42), a biomolecule implicated in the Alzheimer?s disease (AD). In this approach, neurons harvested from embryonic mice were seeded on the substrate-integrated microelectrode arrays. The cultured neurons form a spontaneously active network, and the spiking activity as a functional endpoint could be detected via the MEA. A?42 oligomer, but not monomer, significantly reduced network spike rate. In addition, we demonstrated that the ionotropic glutamate receptors, NMDA and AMPA/kainate, play a role in the effects of A?42 on neuronal activity in vitro. To examine the utility of the MEA-based assay for AD drug discovery, we tested two model therapeutics for AD, methylene blue (MB) and memantine. Our results show an almost full recovery in the activity within 24h after administration of A?42 in the cultures pre-treated with either MB or memantine. Our findings suggest that cultured neuronal networks may be a useful platform in screening potential therapeutics for A? induced changes in neurological function. PMID:26453830

  5. Antiprotozoal and cytotoxic activities in vitro of Colombian Annonaceae.

    PubMed

    Osorio, Edison; Arango, Gabriel Jaime; Jiménez, Nora; Alzate, Fernando; Ruiz, Grace; Gutiérrez, David; Paco, Marco Antonio; Giménez, Alberto; Robledo, Sara

    2007-05-22

    Ethnobotanical and chemotaxonomical studies for antiparasitic activity of Colombian Annonaceae were carried out. In vitro antiprotozoal activity of 36 extracts obtained from six different species was determined against promastigotes of three Leishmania species, epimastigotes of Trypanosoma cruzi and both chloroquine sensitive (F32) and resistant (W2) Plasmodium falciparum. Cytotoxic activity was evaluated in U-937 cells. Active extracts were selected according their selectivity index (SI). Extracts from Annona muricata, Rollinia exsucca, Rollinia pittieri and Xylopia aromatica were active against Leishmania spp. and Trypanosoma cruzi showing IC50 values lower than 25 microg/ml. Hexane extract from Rollinia pittieri leaves was the most selective against Trypanosoma cruzi and Leishmania spp. (IS=10 and 16, respectively). The extracts from Desmopsis panamensis, Pseudomalmea boyacana, Rollinia exsucca and Rollinia pittieri showed good antiplasmodial activity (IC50 < 10 microg/ml). No correlation between antiplasmodial activity and inhibition of beta-hematin production was found. The present study gives specific and useful information about antiprotozoal and cytotoxic activities of some Annonaceae extracts. Results presented here also demonstrate which plants and/or plant parts could be useful in the treatment of leishmaniasis, Chagas' disease and malaria. PMID:17296281

  6. Characterization of the in vitro activity of novel lipoglycopeptide antibiotics.

    PubMed

    Arhin, Francis F; Belley, Adam; McKay, Geoffrey A; Moeck, Gregory

    2010-02-01

    The increasing incidence of antibiotic resistance in human pathogens is of significant concern. Resistance to the widely-used and highly effective glycopeptide antibacterial agent vancomycin, which has been in clinical use for over half a century, has emerged in staphylococci and enterococci. This has spurred the development of newer glycopeptide agents, some of which show activity against vancomycin-resistant organisms. The newer agents currently being developed contain lipophilic side-chains, which distinguish them from vancomycin; as such, they are categorized as lipoglycopeptides. Oritavancin, telavancin, and dalbavancin are lipoglycopeptides in late-stage development to combat Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci. This unit describes methods that may be used to assess the in vitro activities of lipoglycopeptides. The methods include susceptibility assays, time-kill and time-kill-synergy assays, inhibition of synthesis of macromolecules, membrane perturbation assays, and measurement of activity against biofilms. PMID:20131224

  7. Spermicidal activity of Indian seaweeds: an in vitro study.

    PubMed

    Prakash, S; Ravikumar, S; Reddy, K V R; Kannapiran, E

    2014-05-01

    Contraceptive properties of seaweeds are still stands as lacuna; in this context, the screening of in vitro male contraceptive properties of crude ethanolic extract of Indian seaweeds against normal human sperm is carried out. In total, twelve seaweeds were screened for in vitro spermicidal activity. Among these twelve seaweeds, Halimeda gracilis showed 100% inhibition of human spermatozoa at 10 mg ml(-1) concentration in 20 s and its EC50 value was 2.05 mg ml(-1) in 20 s. Further, dose- and time-dependent spermicidal assay revealed that the sperm was completely immobilised for 20 s. Plasma membrane of sperm was damaged due to the exposure of H. gracilis extract. MTT assay with H. gracilis extract showed 88.5% of cytotoxic incidence. H. gracilis extract tested for cytotoxicity against Artemia salina recorded LC50 value of 34.8 ?g ml(-1) . Phytochemical analysis of H. gracilis extract evidenced the presence of alkaloids, flavonoids, proteins and sugars. Results of this study clearly inferred that the synergistic effect of active principles reside within the H. gracilis extract had shown better contraceptive activity. PMID:23557355

  8. In vitro and in vivo activities of antibiotic PM181104.

    PubMed

    Mahajan, Girish; Thomas, Becky; Parab, Rajashri; Patel, Zarine E; Kuldharan, Sandip; Yemparala, Vijayaphanikumar; Mishra, Prabhu Dutt; Ranadive, Prafull; D'Souza, Lisette; Pari, Koteppa; Sivaramkrishnan, H

    2013-11-01

    Drug resistance has become a global threat that, if not addressed, may return us to the preantibiotic era. A way to overcome the problem of growing incidence of global antibiotic resistance is to introduce compounds belonging to classes that are new to the clinic. During a screening of the marine microbial extract library for new antibiotics, one of the extracts showed promising antibacterial activity against Gram-positive organisms. Bioactivity-guided isolation and characterization of active metabolites led to the discovery of a novel thiazolyl cyclic-peptide antibiotic, PM181104. It was isolated and characterized from a marine sponge-associated actinobacterium strain of the genus Kocuria (MTCC 5269). The compound exhibited a potent in vitro antibacterial activity against a broad range of Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). The MIC values evaluated for the compound were found to be in the single-digit nanomolar range. In in vivo studies of PM181104 in a BALB/c murine septicemia model, the compound displayed 100% effective dose (ED100) values of 2.5 and 5.0 mg/kg of body weight against MRSA and 10.0 mg/kg against VRE. In this report, in vitro and in vivo studies of PM181104 are described. PMID:23939903

  9. Spontaneous activity of rat pretectal nuclear complex neurons in vitro

    PubMed Central

    Prochnow, Nora; Schmidt, Matthias

    2004-01-01

    Background Neurons in the mammalian pretectum are involved in the control of various visual and oculomotor tasks. Because functionally independent pretectal cell populations show a wide variation of response types to visual stimulation in vivo, they may also differ in their intrinsic properties when recorded in vitro. We therefore performed whole-cell patch clamp recordings from neurons in the caudal third of the pretectal nuclear complex in frontal brain slices obtained from 3 to 6 week old hooded rats and tried to classify pretectal neurons electrophysiologically. Results Pretectal neurons showed various response types to intracellular depolarizations, including bursting and regular firing behavior. One population of pretectal nuclear complex neurons could be particularly distinguished from others because they displayed spontaneous activity in vitro. These cells had more positive resting potentials and higher input resistances than cells that were not spontaneously active. The maintained firing of spontaneously active pretectal cells was characterized by only small variances in interspike intervals and thus showed a regular temporal patterning. The firing rate was directly correlated to the membrane potential. Removing excitatory inputs by blockade of AMPA and/or NMDA receptors did not change the spontaneous activity. Simultaneous blockade of excitatory and inhibitory synaptic input by a substitution of extracellular calcium with cobalt neither changed the firing rate nor its temporal patterning. Each action potential was preceeded by a depolarizing inward current which was insensitive to calcium removal but which disappeared in the presence of tetrodotoxin. Conclusions Our results indicate that a specific subpopulation of pretectal neurons is capable of generating maintained activity in the absence of any external synaptic input. This maintained activity depends on a sodium conductance and is independent from calcium currents. PMID:15333139

  10. Antifungal Activity of Ellagic Acid In Vitro and In Vivo.

    PubMed

    Li, Zhi-Jian; Guo, Xin; Dawuti, Gulina; Aibai, Silafu

    2015-07-01

    Ellagic acid (EA) has been shown to have antioxidant, antibacterial, and anti-inflammatory activities. In Uighur traditional medicine, Euphorbia humifusa Willd is used to treat fungal diseases, and recent studies suggest that it is the EA content which is responsible for its therapeutic effect. However, the effects of EA on antifungal activity have not yet been reported. This study aimed to investigate the inhibitory effect of EA on fungal strains both in vitro and in vivo. The minimal inhibitory concentration (MIC) was determined by the National Committee for Clinical Laboratory Standards (M38-A and M27-A2) standard method in vitro. EA had a broad spectrum of antifungal activity, with MICs for all the tested dermatophyte strains between 18.75 and 58.33?µg/ml. EA was also active against two Candida strains, with MICs between 25.0 and 75.0?µg/ml. It was inactive against Candida glabrata. The susceptibility of six species of dermatophytes to EA was comparable with that of the commercial antifungal, fluconazole. The most sensitive filamentous species was Trichophyton rubrum (MIC?=?18.75?µg/ml). Studies on the mechanism of action using an HPLC-based assay and an enzyme linked immunosorbent assay showed that EA inhibited ergosterol biosynthesis and reduced the activity of sterol 14?-demethylase P450 (CYP51) in the Trichophyton rubrum membrane, respectively. An in vivo test demonstrated that topical administration of EA (4.0 and 8.0?mg/cm(2) ) significantly enhanced the cure rate in a guinea-pig infection model of Trichophyton rubrum. The results suggest that EA has the potential to be developed as a natural antifungal agent. PMID:25919446

  11. Activity-dependent plasticity of mouse hippocampal assemblies in vitro

    PubMed Central

    Keller, Martin K.; Draguhn, Andreas; Both, Martin; Reichinnek, Susanne

    2015-01-01

    Memory formation is associated with the generation of transiently stable neuronal assemblies. In hippocampal networks, such groups of functionally coupled neurons express highly ordered spatiotemporal activity patterns which are coordinated by local network oscillations. One of these patterns, sharp wave-ripple complexes (SPW-R), repetitively activates previously established groups of memory-encoding neurons, thereby supporting memory consolidation. This function implies that repetition of specific SPW-R induces plastic changes which render the underlying neuronal assemblies more stable. We modeled this repetitive activation in an in vitro model of SPW-R in mouse hippocampal slices. Weak electrical stimulation upstream of the CA3-CA1 networks reliably induced SPW-R of stereotypic waveform, thus representing re-activation of similar neuronal activity patterns. Frequent repetition of these patterns (100 times) reduced the variance of both, evoked and spontaneous SPW-R waveforms, indicating stabilization of pre-existing assemblies. These effects were most pronounced in the CA1 subfield and depended on the timing of stimulation relative to spontaneous SPW-R. Additionally, plasticity of SPW-R was blocked by application of a NMDA receptor antagonist, suggesting a role for associative synaptic plasticity in this process. Thus, repetitive activation of specific patterns of SPW-R causes stabilization of memory-related networks. PMID:26041998

  12. In Vitro Antimicrobial and Antiproliferative Activity of Amphipterygium adstringens

    PubMed Central

    Rodriguez-Garcia, A.; Peixoto, I. T. A.; Verde-Star, M. J.; De la Torre-Zavala, S.; Aviles-Arnaut, H.; Ruiz, A. L. T. G.

    2015-01-01

    Amphipterygium adstringens is a plant widely used in Mexican traditional medicine for its known anti-inflammatory and antiulcer properties. In this work, we evaluated the in vitro antimicrobial and antiproliferative activities of the methanolic extract of A. adstringens against oral pathogens such as Streptococcus mutans, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Candida albicans, and Candida dubliniensis, using microdilution (MIC) and agar diffusion methods (MBC), and the antiproliferative activity evaluating total growth inhibition (TGI) by staining the protein content with sulforhodamine B (SRB), using nine human cancer cell lines. Crude extract (CE) of A. adstringens showed some degree of activity against one or more of the strains with a MIC from 0.125?mg/mL to 63?mg/mL and MBC from 1.6 to 6.3?mg/mL and cytotoxic activity, particularly against NCI-ADR/RES, an ovarian cell line expressing multiple resistance drugs phenotype. The CE is a complex mixture of possible multitarget metabolites that could be responsible for both antimicrobial and antiproliferative activities, and further investigation is required to elucidate the identity of active compounds. Nevertheless the CE itself is useful in the development of new antimicrobial treatment based on natural products to prevent oral diseases and as alternative natural source for cancer treatment and prevention. PMID:26451151

  13. In Vitro Antimicrobial and Antiproliferative Activity of Amphipterygium adstringens.

    PubMed

    Rodriguez-Garcia, A; Peixoto, I T A; Verde-Star, M J; De la Torre-Zavala, S; Aviles-Arnaut, H; Ruiz, A L T G

    2015-01-01

    Amphipterygium adstringens is a plant widely used in Mexican traditional medicine for its known anti-inflammatory and antiulcer properties. In this work, we evaluated the in vitro antimicrobial and antiproliferative activities of the methanolic extract of A. adstringens against oral pathogens such as Streptococcus mutans, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Candida albicans, and Candida dubliniensis, using microdilution (MIC) and agar diffusion methods (MBC), and the antiproliferative activity evaluating total growth inhibition (TGI) by staining the protein content with sulforhodamine B (SRB), using nine human cancer cell lines. Crude extract (CE) of A. adstringens showed some degree of activity against one or more of the strains with a MIC from 0.125?mg/mL to 63?mg/mL and MBC from 1.6 to 6.3?mg/mL and cytotoxic activity, particularly against NCI-ADR/RES, an ovarian cell line expressing multiple resistance drugs phenotype. The CE is a complex mixture of possible multitarget metabolites that could be responsible for both antimicrobial and antiproliferative activities, and further investigation is required to elucidate the identity of active compounds. Nevertheless the CE itself is useful in the development of new antimicrobial treatment based on natural products to prevent oral diseases and as alternative natural source for cancer treatment and prevention. PMID:26451151

  14. In vitro Antioxidant Activities of Trianthema portulacastrum L. Hydrolysates

    PubMed Central

    Yaqoob, Sadaf; Sultana, Bushra; Mushtaq, Muhammad

    2014-01-01

    Hydrolysates of Trianthema portulacastrum in acidified methanol were evaluated for their total phenolic (TP) constituents and respective antioxidant activities using in vitro assays (i.e., 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, percent inhibition of linoleic acid peroxidation, and ferric reducing power). The observed results indicate that root, shoot, and leaf fractions of T. portulacastrum contain 50.75~98.09 mg gallic acid equivalents/g dry weight of TP. In addition, these fractions have substantial reducing potentials (0.10~0.59), abilities to inhibit peroxidation (43.26~89.98%), and DPPH radical scavenging capabilities (6.98~311.61 ?g/mL IC50). The experimental data not only reveal T. portulacastrum as potential source of valuable antioxidants, but also indicate that acidified methanol may be an ideal choice for the enhanced recovery of phenolic compounds with retained biological potential for the food and pharmaceutical industry. PMID:24772406

  15. Australian plants show anthelmintic activity toward equine cyathostomins in vitro.

    PubMed

    Payne, S E; Kotze, A C; Durmic, Z; Vercoe, P E

    2013-09-01

    Anthelmintic resistance in gastrointestinal parasites of horses is an increasing problem, particularly in cyathostomins, and there is a need to find alternative means for the control of these parasites. We screened crude extracts from 37 species of Australian native plants for their anthelmintic activity in vitro against cyathostomin larvae (development from egg to third larval stage), with the aim of identifying those species that may be suitable for incorporation into sustainable parasite management programs. Water extracts from seven species, namely Acacia baileyana, Acacia melanoxylon, Acacia podalyriifolia, Alectryon oleifolius, Duboisia hopwoodii, Eucalyptus gomphocephala and Santalum spicatum completely inhibited larval development (100% inhibition compared to the control), while another 10 species caused 90% inhibition at the initial screening concentration of 1400 ?g of extractable solids/mL. The seven most potent extracts produced IC50 values (concentration of extract which resulted in a 50% inhibition of development) in the range 30.9-196 ?g/mL. Fourteen extracts were incubated with polyvinylpolypyrrolidone (PVPP) before the assays, which removed the anthelmintic activity from 12 of these extracts, indicating that tannins were likely to be the bioactive compound responsible for the effect, while in two species, i.e. A. melanoxylon and D. hopwoodii, compounds other than tannins were likely to be responsible for their anthelmintic action. Our results suggest that a number of Australian native plants have significant anthelmintic activity against cyathostomin larval development in vitro. There is potential for these plants to be used as part of sustainable parasite control programs in horses, although more research is needed to identify the compounds responsible for the anthelmintic effects and confirm their activity in vivo. PMID:23394801

  16. In Vitro Anti-rotaviral Activity of Achillea kellalensis

    PubMed Central

    Taherkhani, Reza; Farshadpour, Fatemeh; Makvandi, Manoochehr

    2013-01-01

    Background Achillea kellalensis, which is frequently used by Chaharmahal va Bakhtiarians residing in, Southwest of Iran, as a traditional herbal medicine for the treatment of acute diarrhea, has been selected to examine its antiviral activities against bovine rotavirus and cell toxicity activity in MA-104 cells. Objectives The aim of this study was to evaluate the in vitro cytotoxic and anti-rotavirus properties of crude extracts of A. kellalensis. Materials and Methods The dried and powdered flowers of Achillea kellalensis were extracted with hot water and ethanol 50% (v/v). The cell viability and toxicity of the extracts were evaluated on MA-104 cells using four methods; trypan blue dye, NR, crystal violet and MTT assay. The in vitro anti-rotavirus properties were determined via four different assays, in order to evaluate the direct inhibition and/or the inhibition of viral replication. Results Cytotoxicity of two A. kellalensis extracts showed different concentrations. Hydro-alcoholic extract had low CC50 at 600 µg/mL by the NR assay while the aqueous extract had high CC50 at 1000µg/mL by the crystal violet method. In the simultaneous treatment assay and post treatment assay, the extracts were able to prevent viral replication and inhibit the viral CPE on MA-104 cells at 10 TCID50, but the extracts did not exhibit direct antiviral activity on rotavirus adsorption. The effective concentration (EC50) of both extracts was observed to be 100 µg/mL. Conclusions These results indicate that A. kellalensis extracts exert potent anti-rotaviral activity only after viral adsorption. The two extracts from A. kellalensis showed a good selectivity index. Also these results suggest that extracts prepared from the flowers of A. kellalensis may be potential anti-rotaviral agents in vivo and be useful in veterinary medicine. PMID:24624203

  17. In Vitro Activity of Ceftazidime-Avibactam Combination in In Vitro Checkerboard Assays

    PubMed Central

    Melchers, Maria J.; van Mil, Anita C.; Nichols, Wright W.

    2014-01-01

    To evaluate the in vitro effects of the combination of ceftazidime and avibactam on the MICs of both compounds, checkerboard assays were performed for 81 clinical strains, including 55 Enterobacteriaceae strains (32 Klebsiella pneumoniae, 19 Escherichia coli, 1 Citrobacter freundii, and 3 Enterobacter cloacae) and 26 strains of Pseudomonas aeruginosa, all with known resistance mechanisms such as extended-spectrum ?-lactamases (ESBLs) and carbapenemases, phenotypically or molecularly determined. Phenotypically ceftazidime-resistant strains (n = 69) were analyzed in more detail. For the Enterobacteriaceae strains, a concentration-dependent effect of avibactam was found for most strains with a maximum effect of avibactam at a concentration of 4 mg/liter, which decreased all ceftazidime MICs to ?4 mg/liter. Avibactam alone also showed antibacterial activity (the MIC50 and MIC90 being 8 and 16 mg/liter, respectively). For the ceftazidime-resistant P. aeruginosa strains, considerable inhibition of ?-lactamases by avibactam was acquired at a concentration of 4 mg/liter, which decreased all ceftazidime MICs except one to ?8 mg/liter (the CLSI and EUCAST susceptible breakpoint). Increasing the concentration of avibactam further decreased the MICs, resulting in a maximum effect for most strains at 8 to 16 mg/liter. In summary, for most strains, the tested addition of avibactam of 4 mg/liter restored the antibacterial activity of ceftazidime to a level comparable to that of wild-type strains, indicating full inhibition, and strains became susceptible according to the EUCAST and CLSI criteria. Based on these in vitro data, avibactam is a promising inhibitor of different ?-lactamases, including ESBLs and carbapenemases. PMID:25487794

  18. In Vitro Antileukemic Activity of Xanthosoma sagittifolium (Taioba) Leaf Extract

    PubMed Central

    Caxito, Marina L. C.; Correia, Rachell R.; Gomes, Anne Caroline C.; Justo, Graça; Coelho, Marsen G. P.; Sakuragui, Cássia M.; Kuster, Ricardo M.; Sabino, Katia C. C.

    2015-01-01

    Xanthosoma sagittifolium Schott is a herb of the Araceae family, popularly known as taioba, which is consumed as food in some regions of Brazil, Africa, and Asia. This species has already been evaluated for the antifungal activities. However, based on its potential antitumor activity, the present study further aimed to examine the antitumor, as well as chelation, activity of X. sagittifolium leaf extract. Results showed that hydroethanolic extract of X. sagittifolium leaves (HEXs-L) exhibits cytotoxic effects against the immortalized line of human T-lymphocytic (Jurkat) and myelogenous (K562) leukemia cells, but not nontumor RAW 264.7 macrophages or NIH/3T3 fibroblasts. HEXs-L inhibited 50.3% of Jurkat cell proliferation, reducing by 20% cells in G2/M phase, but increasing cells in sub-G1 phase, thereby inducing apoptosis by 54%. In addition, HEXs-L inhibited NO production by 59%, as determined by Griess reaction, and chelated 93.8% of free Fe(II), as demonstrated by ferrozine assay. Phytochemical studies were carried out by ESI-MS, identifying apigenin di-C-glycosides as major compounds. Overall, this work revealed that leaf extract of Xanthosoma sagittifolium presented chelating activity and in vitro antitumor activity, arresting cell cycle and inducing apoptosis of leukemia cells, thus providing evidence that taioba leaves may have practical application in cancer therapy. PMID:26180533

  19. Action of T-activin on activity of human natural killer cells in vitro

    SciTech Connect

    Cheknev, S.B.; Saidov, M.Z.; Koval'chuk, L.V.; Pavlyuk, A.S.; Arion, V.Ya.

    1986-09-01

    This paper describes a study of the action of T-activin on activity of human natural killer cells (NKC) in vitro. The K-562 chronic human myeloid leukemia cells, cultured in vitro, used as targets were labeled with /sup 3/H-uridine. The experimental results indicate that T-activin can depress NKC activity but under certain conditions, it can also stimulate NKC. T-activin possesses immunoregulatory properties relative to NKC activity in vitro.

  20. In-vitro Activity of Avermectins against Mycobacterium ulcerans

    PubMed Central

    Omansen, Till F.; Porter, Jessica L.; Johnson, Paul D. R.; van der Werf, Tjip S.; Stienstra, Ymkje; Stinear, Timothy P.

    2015-01-01

    Mycobacterium ulcerans causes Buruli ulcer (BU), a debilitating infection of subcutaneous tissue. There is a WHO-recommended antibiotic treatment requiring an 8-week course of streptomycin and rifampicin. This regime has revolutionized the treatment of BU but there are problems that include reliance on daily streptomycin injections and side effects such as ototoxicity. Trials of all-oral treatments for BU show promise but additional drug combinations that make BU treatment safer and shorter would be welcome. Following on from reports that avermectins have activity against Mycobacterium tuberculosis, we tested the in-vitro efficacy of ivermectin and moxidectin on M. ulcerans. We observed minimum inhibitory concentrations of 4–8 ?g/ml and time-kill assays using wild type and bioluminescent M. ulcerans showed a significant dose-dependent reduction in M. ulcerans viability over 8-weeks. A synergistic killing-effect with rifampicin was also observed. Avermectins are well tolerated, widely available and inexpensive. Based on our in vitro findings we suggest that avermectins should be further evaluated for the treatment of BU. PMID:25742173

  1. Anti-hepatoma activity of resveratrol in vitro

    PubMed Central

    Sun, Zhong-Jie; Pan, Cheng-En; Liu, Hong-Shan; Wang, Guo-Jun

    2002-01-01

    AIM: To study the anti-tumor effect of resveratrol alone and the synergistic effects of resveratrol with 5-FU on the growth of H22 cells line in vitro METHODS: The number of cells was measured by MTT method,the morphological changes of H22 cells were investigated under microscopy and electron microscopyq. RESULTS: Resveratrol inhibited the growth of hepatoma cells line H22 in a dose- and time-dependent manner. IC50 of the resveratrol on H22 cells was 6.57 mg·L-1. The synergistic anti-tumor effects of resveratrol with 5-FU increased to a greater extent than for H22 cells treated with 5-FU alone (70.2% vs 28.4%)(P < 0.05). Under microscope and electron microscope, characteristics of apoptosis such as typical apoptotic bodies were commonly found in tumor cells in the drug-treated groups. CONCLUSION: Resveratrol can suppresses the growth of H22 cells in vitro, its anti-tumor activity may occur through the induction of apoptosis. PMID:11833076

  2. Standardization and in vitro antioxidant activity of jatamansi rhizome

    PubMed Central

    Singh, Mhaveer; Khan, Mohammad A.; Khan, Masood S.; Ansari, S. H.; Ahmad, Sayeed

    2015-01-01

    Background: Nardostachys jatamansi Linn. commonly known as jatamansi is a well notorious drug in Indian systems of medicines having various health-related benefits and employed in various herbal formulations due to the presence of high levels of valuable phenolic constituents. The present study was aimed to quality assessment of Jatamansi rhizome by studying macro- and micro-scopic characters along with physicochemical tests, chemo-profiling using thin layer chromatography (TLC), and gas chromatography–mass spectrometry (GC-MS), in vitro antioxidant activity. Materials and Methods: Standardization was carried out as per the pharmacopeial guidelines and contaminant estimation was carried out by analyzing the samples for the determination of heavy metals, pesticides, and aflatoxins. Chemo-profiling was done with TLC by optimizing the mobile phase for different extracts. The GC-MS chemo-profiling was also carried out by using hexane soluble fraction of the hydroalcoholic extract. The drug is well known for a protective role in the human body as an antioxidant, so total phenolic contents and in vitro antioxidant efficacy was also determined by using established methods. Results: The results of quality control and anatomical studies were very much useful for its identification, whereas significant antioxidant efficacy was also observed. The drug was found free of contaminants when analyzed for pesticides and aflatoxins, whereas heavy metals were found under the pharmacopeial limit. Conclusion: The findings of the present research can be utilized for the identification and quality control of the jatamansi rhizome. PMID:26681882

  3. In Vitro Activity of Daptomycin in Combination with -Lactams, Gentamicin, Rifampin, and Tigecycline against Daptomycin-

    E-print Network

    Nizet, Victor

    In Vitro Activity of Daptomycin in Combination with -Lactams, Gentamicin, Rifampin, and Tigecycline of daptomycin in combination with ampi- cillin, cefazolin, ceftriaxone, ceftaroline, ertapenem, gentamicin

  4. In vitro radical scavenging activity of two Columbian Magnoliaceae

    NASA Astrophysics Data System (ADS)

    Puertas M., Miguel A.; Mesa v., Ana M.; Sáez v., Jairo A.

    2005-08-01

    The recent interest in the conservation of the tropical forest is due, at least in part, to the potential economic and health benefits that can be exploited from several plants. This report shows the in vitro antioxidant activity of some fractions isolated from leaves of two Columbian Magnoliaceae, Talauma hernandezii G. Lozano-C and Dugandiodendron yarumalense Lozano. The activity was determined using the radical monocation 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS·+) and the stable free radical 2-2-diphenyl-1-picrylhydrazyl (DPPH·), as part of general biological screening of these plants. The antioxidant capacity obtained from fractions was similar to those of ?-tocopherol, tert-butylated hydroxyanisole (BHA), and ascorbic acid. The most active scavenger extract was the fraction 7 (TAA = 48.6 mmol Trolox/kg extract and IC50 ? 0.01 kg extract/mmol DPPH); and the least active was the fraction 1 (TAA = 11.23 mmol Trolox/kg extract and IC50 = 0.21 kg extract/mmol DPPH) all of them isolated from D. yarumalense. These results suggest that these plants can be attractive as source of antioxidant compounds with the ability to reduce radicals like ATBS and DPPH.

  5. Silver and Gold Nanoparticles Alter Cathepsin Activity In vitro

    PubMed Central

    2011-01-01

    Nanomaterials are being incorporated into many biological applications for use as therapeutics, sensors, or labels. Silver nanomaterials are being utilized for biological implants and wound dressings as an antiviral material, whereas gold nanomaterials are being used as biological labels or sensors due to their surface properties and biocompatibility. Cytotoxicity data of these materials are becoming more prevalent; however, little research has been performed to understand how the introduction of these materials into cells affects cellular processes. Here, we demonstrate the impact that silver and gold nanoparticles have on cathepsin activity in vitro. Cathepsins are important cellular proteases that are imperative for proper immune system function. We have selected to examine gold and silver nanoparticles due to the increased use of these materials in biological applications. This manuscript depicts how both of these types of nanomaterials affect cathepsin activity, which could impact the host's immune system and its ability to respond to pathogens. Cathepsin B activity decreases in a dose-dependent manner with all nanoparticles tested. Alternatively, the impact of nanoparticles on cathepsin L activity depends greatly on the type and size of the material.

  6. Phosphine derivatives of sparfloxacin - Synthesis, structures and in vitro activity

    NASA Astrophysics Data System (ADS)

    Komarnicka, Urszula K.; Starosta, Rados?aw; Guz-Regner, Katarzyna; Bugla-P?osko?ska, Gabriela; Kyzio?, Agnieszka; Je?owska-Bojczuk, Ma?gorzata

    2015-09-01

    We synthesized two derivatives of sparfloxacin (HSf): aminomethyl(diphenyl)phosphine (PSf) and its oxide (OPSf). The compounds were characterized by NMR spectroscopy, MS and elemental analysis. In addition, the molecular structures of the compounds were determined using DFT and X-ray (OPSf) analysis. The antibacterial activity of HSf and both derivatives was tested against four reference and fifteen clinical Gram-positive and Gram-negative strains of bacteria (sensitive or resistant to fluoroquinolones). The results showed that the activity of PSf was similar to or higher than the activity of HSf, while OPSf was found significantly less active. The compounds were also tested in vitro toward the following cancer cell lines: mouse colon carcinoma (CT26) and human lung adenocarcinoma (A549). Regardless of the cancer cell line, derivatization of HSf resulted in the gradual increase of cytotoxicity. OPSf exhibited the highest one (4 h - incubation time: IC50(CT26) = 51.0 ± 1.2; IC50(A549) = 74.9 ± 1.4 and 24 h: IC50(CT26) = 109.2 ± 8.8; IC50(A549) = 52.7 ± 9.2).

  7. In Vitro Activities of Hexaazatrinaphthylenes against Leishmania spp.

    PubMed Central

    García-Velázquez, Daniel; Martín-Navarro, Carmen M.; Sifaoui, Ines; Reyes-Batlle, María; Lorenzo-Morales, Jacob; Gutiérrez-Ravelo, Ángel; Pińero, José E.

    2015-01-01

    The in vitro activity of a novel group of compounds, hexaazatrinaphthylene derivatives, against two species of Leishmania is described in this study. These compounds showed a significant dose-dependent inhibition effect on the proliferation of the parasites, with 50% inhibitory concentrations (IC50s) ranging from 1.23 to 25.05 ?M against the promastigote stage and 0.5 to 0.7 ?M against intracellular amastigotes. Also, a cytotoxicity assay was carried out to in order to evaluate the possible toxic effects of these compounds. Moreover, different assays were performed to determine the type of cell death induced after incubation with these compounds. The obtained results highlight the potential use of hexaazatrinaphthylene derivatives against Leishmania species, and further studies should be undertaken to establish them as novel leishmanicidal therapeutic agents. PMID:25753635

  8. Lethal Factor Active-Site Mutations Affect Catalytic Activity In Vitro

    PubMed Central

    Hammond, S. E.; Hanna, P. C.

    1998-01-01

    The lethal factor (LF) protein of Bacillus anthracis lethal toxin contains the thermolysin-like active-site and zinc-binding consensus motif HEXXH (K. R. Klimpel, N. Arora, and S. H. Leppla, Mol. Microbiol. 13:1093–1100, 1994). LF is hypothesized to act as a Zn2+ metalloprotease in the cytoplasm of macrophages, but no proteolytic activities have been previously shown on any target substrate. Here, synthetic peptides are hydrolyzed by LF in vitro. Mass spectroscopy and peptide sequencing of isolated cleavage products separated by reverse-phase high-pressure liquid chromatography indicate that LF seems to prefer proline-containing substrates. Substitution mutations within the consensus active-site residues completely abolish all in vitro catalytic functions, as does addition of 1,10-phenanthroline, EDTA, and certain amino acid hydroxamates, including the novel zinc metalloprotease inhibitor ZINCOV. In contrast, the protease inhibitors bestatin and lysine CMK, previously shown to block LF activity on macrophages, did not block LF activity in vitro. These data provide the first direct evidence that LF may act as an endopeptidase. PMID:9573135

  9. Evaluation of the In Vitro Antifungal Activity of Allicin

    PubMed Central

    Yamada, Yasuo; Azuma, Keiz?

    1977-01-01

    Allicin was effective in vitro against Candida, Cryptococcus, Trichophyton, Epidermophyton, and Microsporum. The minimal inhibitory concentrations (MICs) of allicin against these organisms were 3.13 to 6.25 ?g/ml by the agar dilution method and 1.57 to 6.25 ?g/ml by the broth dilution method, using Sabouraud glucose (SG) medium. However decreased activity was demonstrated against Aspergillus. The MIC of allicin against various pathogenic fungi was affected considerably by differences in the experimental conditions, e.g., incubation time, inoculum size, type of medium, and medium pH. The MIC of allicin against Candida, Cryptococcus, and Aspergillus remained constant after more than 3 days of incubation, and that against Dermatophytes remained constant after more than 10 days of incubation. Decreasing the inoculum size increased the susceptibility to allicin. The antifungal activity of allicin was stronger on SG agar medium with a pH of 5.6 than on the same medium with a pH of 6.0 or higher. By microscopical observation, allicin induced morphological abnormalities in hyphae of Trichophyton mentagrophytes Morita. Percent germination of spores of the Morita strain at 24 h in SG agar medium was greatly decreased with an allicin concentration of 3.13 ?g/ml, and the lethal dose for the spores was about four times higher than the fungistatic concentration. These results suggest that allicin inhibits both germination of spores and growth of hyphae. Images PMID:856026

  10. In vitro activity of the new quinolone BAY y 3118 against anaerobic bacteria.

    PubMed

    Nord, C E; Lindmark, A; Persson, I

    1993-08-01

    The in vitro activity of BAY y 3118 against anaerobic cocci, Propionibacterium acnes, Clostridium perfringens, Clostridium difficile, Bacteroides fragilis, other Bacteroides spp. and fusobacteria was determined by an agar dilution method. This activity was compared with that of ciprofloxacin, ofloxacin, piperacillin, cefoxitin, imipenem, clindamycin and metronidazole. BAY y 3118, imipenem, clindamycin and metronidazole were the most active agents tested. The in vitro activity of BAY y 3118 against anaerobic bacteria was superior to that of ciprofloxacin and ofloxacin. PMID:8223667

  11. In vitro antimicrobial activity of Medilox® super-oxidized water

    PubMed Central

    2014-01-01

    Aim Super-oxidized water is one of the broad spectrum disinfectants, which was introduced recently. There are many researches to find reliable chemicals which are effective, inexpensive, easy to obtain and use, and effective for disinfection of microorganisms leading hospital infections. Antimicrobial activity of super-oxidized water is promising. The aim of this study was to investigate the in-vitro antimicrobial activity of different concentrations of Medilox® super-oxidized water that is approved by the Food and Drug Administration (FDA) as high level disinfectant. Material and methods In this study, super-oxidized water obtained from Medilox® [Soosan E & C, Korea] device, which had been already installed in our hospital, was used. Antimicrobial activities of different concentrations of super-oxidized water (1/1, 1/2, 1/5, 1/10, 1/20, 1/50, 1/100) at different exposure times (1, 2, 5, 10, 30 min) against six ATCC strains, eight antibiotic resistant bacteria, yeasts and molds were evaluated using qualitative suspension test. Dey-Engley Neutralizing Broth [Sigma-Aldrich, USA] was used as neutralizing agent. Results Medilox® was found to be effective against all standard strains (Acinetobacter baumannii 19606, Escherichia coli 25922, Enterococcus faecalis 29212, Klebsiella pneumoniae 254988, Pseudomonas aeruginosa 27853, Staphylococcus aureus 29213), all clinical isolates (Acinetobacter baumannii, Escherichia coli, vancomycin-resistant Enterococcus faecium, Klebsiella pneumoniae, Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, Bacillus subtilis, Myroides spp.), and all yeastsat 1/1 dilution in ??1 minute. It was found to be effective on Aspergillus flavus at 1/1 dilution in ??2 minutes and on certain molds in ??5 minutes. Conclusion Medilox® super-oxidized water is a broad spectrum, on-site producible disinfectant, which is effective on bacteria and fungi and can be used for the control of nosocomial infection. PMID:25023905

  12. In vitro antioxidant activity of Diospyros malabarica Kostel bark.

    PubMed

    Mondal, Susanta Kumar; Chakraborty, Goutam; Gupta, M; Mazumder, U K

    2006-01-01

    Antioxidant activity of defatted methanol extract of D. malabarica bark was studied for its free radical scavenging property on different in vitro models e.g. 1,1-diphenyl-2-picryl hydrazyl (DPPH), nitric oxide, superoxide, hydroxyl radical and lipid peroxide radical model. The extract showed good dose-dependent free radical scavenging property in all the models except in hydroxyl radical inhibition assay. IC50 values were found to be 9.16, 13.21, 25.27 and 17.33 microg/ml respectively in DPPH, nitric oxide, superoxide and lipid peroxidation inhibition assays. In hydroxyl radical inhibition assay 1000 microg/ml extract showed only 10% inhibition with respect to the control. Measurement of total phenolic compounds by Folin-Ciocalteu's phenol reagent indicated that 1 mg of the extract contained 120.7 microg equivalent of pyrocatechol. The results indicate that the antioxidant property of the extract may be due to the high content of phenolic compounds. However, the underlying mechanism may not involve hydroxyl radical scavenging property. PMID:16430089

  13. Acaricide activity in vitro of Acmella oleracea against Rhipicephalus microplus.

    PubMed

    Castro, K N C; Lima, D F; Vasconcelos, L C; Leite, J R S A; Santos, R C; Paz Neto, A A; Costa-Júnior, L M

    2014-10-01

    Cattle tick control has been limited by the resistance of these parasites to synthetic acaricides. Natural products are a possible alternative as they have different mechanisms of action. Acmella oleracea is a native plant with a large cultivated area in the Amazon region and could be easily used for large-scale preparation of a commercial product. This study evaluated the in vitro action of the hexane extract of the aerial parts of A. oleracea on larvae and engorged females of the cattle tick Rhipicephalus microplus. Spilanthol was the major constituent with a content of 14.8% in the extract. The hexane extract of A. oleracea was highly effective against larvae of R. microplus with an LC50 of 0.8 mg mL(-1). Against engorged females, hexane extract of A. oleracea reduced oviposition and hatchability of eggs with an LC50 of 79.7 mg mL(-1). Larvae and engorged females were killed by the hexane extract with high efficiency (>95%) at concentrations of 3.1 and 150.0 mg mL(-1), respectively. These results demonstrate that the hexane extract of A. oleracea has significant activity against R. microplus and has potential to be developed into formulations for tick control. PMID:25033813

  14. In Vitro Antibacterial Activity of NB-003 against Propionibacterium acnes?

    PubMed Central

    Pannu, J.; McCarthy, A.; Martin, A.; Hamouda, T.; Ciotti, S.; Ma, L.; Sutcliffe, J.; Baker, J. R.

    2011-01-01

    NB-003 and NB-003 gel formulations are oil-in-water nanoemulsions designed for use in bacterial infections. In vitro susceptibility of Propionibacterium acnes to NB-003 formulations and comparator drugs was evaluated. Both NB-003 formulations were bactericidal against all P. acnes isolates, including those that were erythromycin, clindamycin, and/or tetracycline resistant. In the absence of sebum, the MIC90s/minimum bactericidal concentrations (MBC90s) for NB-003, NB-003 gel, salicylic acid (SA), and benzoyl peroxide (BPO) were 0.5/2.0, 1.0/2.0, 1,000/2,000, and 50/200 ?g/ml, respectively. In the presence of 50% sebum, the MIC90s/MBC90s of NB003 and BPOs increased to 128/1,024 and 400/1,600 ?g/ml, respectively. The MIC90s/MBC90s of SA were not significantly impacted by the presence of sebum. A reduction in the MBC90s for NB-003 and BPO was observed when 2% SA or 0.5% BPO was integrated into the formulation, resulting in MIC90s/MBC90s of 128/256 ?g/ml for NB003 and 214/428 ?g/ml for BPO. The addition of EDTA enhanced the in vitro efficacy of 0.5% NB-003 in the presence or absence of 25% sebum. The addition of 5 mM EDTA to each well of the microtiter plate resulted in a >16- and >256-fold decrease in MIC90 and MBC90, yielding a more potent MIC90/MBC90 of ?1/<1 ?g/ml. The kinetics of bactericidal activity of NB-003 against P. acnes were compared to those of a commercially available product of BPO. Electron micrographs of P. acnes treated with NB-003 showed complete disruption of bacteria. Assessment of spontaneous resistance of P. acnes revealed no stably resistant mutant strains. PMID:21746943

  15. Processed tart cherry products--comparative phytochemical content, in vitro antioxidant capacity and in vitro anti-inflammatory activity.

    PubMed

    Ou, Boxin; Bosak, Kristen N; Brickner, Paula R; Iezzoni, Dominic G; Seymour, E Mitchell

    2012-05-01

    Processing of fruits and vegetables affects their phytochemical and nutrient content. Tart cherries are commercially promoted to possess antioxidant and anti-inflammatory activity. However, processing affects their phytochemical content and may affect their related health benefits. The current study compares the in vitro antioxidant capacity and anti-inflammatory cyclooxygenase activity of processed tart cherry (Prunus cerasus) products-cherry juice concentrate, individually quick-frozen cherries, canned cherries, and dried cherries. Cherry products were analyzed for total anthocyanin and proanthocyanidin content and profile. On a per serving basis, total anthocyanins were highest in frozen cherries and total proanthocyanidins were highest in juice concentrate. Total phenolics were highest in juice concentrate. Juice concentrate had the highest oxygen radical absorbance capacity (ORAC) and peroxynitrite radical averting capacity (NORAC). Dried cherries had the highest hydroxyl radical averting capacity (HORAC) and superoxide radical averting capacity (SORAC). Processed tart cherry products compared very favorably to the U.S. Dept. of Agriculture-reported ORAC of other fresh and processed fruits. Inhibition of in vitro inflammatory COX-1 activity was greatest in juice concentrate. In summary, all processed tart cherry products possessed antioxidant and anti-inflammatory activity, but processing differentially affected phytochemical content and in vitro bioactivity. On a per serving basis, juice concentrate was superior to other tart cherry products. PMID:23163942

  16. In vitro and in vivo activity of 3-alkoxy-1,2-dioxolanes against Schistosoma mansoni

    PubMed Central

    Ingram, Katrin; Schiaffo, Charles E.; Sittiwong, Wantanee; Benner, Evan; Dussault, Patrick H.; Keiser, Jennifer

    2012-01-01

    Objectives Compounds characterized by a peroxidic skeleton are an interesting starting point for antischistosomal drug discovery. Previously a series of 3-alkoxy-1,2-dioxolanes, which are chemically stable cyclic peroxides, demonstrated significant in vitro activity against Plasmodium falciparum. We aimed to evaluate the potential of these compounds against Schistosoma mansoni and elucidate the roles of iron and peroxidic groups in activity. Methods Drugs were tested against juvenile and adult stages of S. mansoni in vitro and in vivo. Selected structures were assessed in vitro against schistosomes in the presence of additional iron sources. In addition, drugs were tested in vitro and in vivo against Echinostoma caproni, a non-blood-feeding intestinal fluke. Finally, the activity of non-peroxidic analogues was evaluated. Results Three dioxolanes displayed IC50s ?20.1 ?M against adult schistosomes and values as low as 4.2 ?M against newly transformed schistosomula. Nonetheless, only moderate, non-significant worm burden reductions were observed after treatment of mice harbouring adult infections. Drugs lacked activity against juvenile schistosomes in vivo. Two selected dioxolanes showed in vitro activity against E. caproni down to concentrations of 5 mg/L, but none of the compounds revealed in vivo activity. All tested non-peroxidic analogues lacked activity in vitro against both parasites. Conclusions Selected dioxolanes presented interesting in vitro activity, but low in vivo activities have to be overcome to identify a lead candidate. Although the inactivity of non-peroxidic analogues underlines the necessity of a peroxide functional group, incubation of adult schistosomes with additional iron sources did not alter activity, supporting an iron-independent mode of activation. PMID:22553141

  17. Antioxidant, Antibacterial and Antischistosomal Activities of Extracts from Grateloupia livida (Harv). Yamada

    PubMed Central

    Yao, Fen; Chen, Weizhou; Zhong, Shuping; Zheng, Fuchun; Shi, Ganggang

    2013-01-01

    The present study was designated to evaluate the antioxidant, antibacterial and antischistosomal activities of Grateloupia livida (GL) extracts in vitro. A GL Ethanol extract (EE) was separated into petroleum ether (PE), ethyl acetate (EA), n-butyl alcohol (BuOH) and aqueous (AQ) fractions to fractionate the polar and non-polar compounds in the EE. Extracts antioxidant activities were evaluated in vitro by DPPH radical-scavenging, deoxyribose radical scavenging, and ?-carotene bleaching assays, all using butylated hydroxytoluene (BHT) as the reference antioxidant compound. The most effective antioxidant properties were observed in the PE fraction in all three assays. Antimicrobial testing showed that the PE fraction exhibited broad-spectrum antimicrobial activity, with the PE fraction also exhibiting strong activity against the human pathogenic trematode S. japonicum adult worm. In order to investigate the relationships between bioactivity and chemical composition, the chemical composition of the PE fraction was analyzed by gas chromatography-mass spectrometry (GC-MS). In total, 25 components were identified in the PE fraction, most of which have known antioxidant and antimicrobial activities. However, none of the compounds have reported activity against Schistosoma, suggesting that the schistosomicidal activity of the PE fraction may be related to minor constituents present in the extract, or governed by more intricate synergistic or additive relationships. Finally, fractions with the greatest biological activity displayed neither cellular cytotoxicity, at concentrations up to 100 ug/ml, or acute oral toxicity in mice, at doses up to 2000 mg/kg. Based on antioxidant, antimicrobial, antischistosomal activities, and low toxicity, the PE fraction possesses properties useful for food preservation and overall improvement of human health. PMID:24312216

  18. In vitro antioxidant activity of polysaccharide from Gardenia jasminoides ellis

    USGS Publications Warehouse

    Fan, Y.; Ge, Z.; Luo, A.

    2011-01-01

    A water-soluble polysaccharide, GP, was isolated from Gardenia jasminoides Ellis through hot water extraction followed by ethanol precipitation. The in vitro free radicals scavenging tests exhibited that GP has significant scavenging abilities especially for ABTS, DPPH, and hydroxyl radicals, which suggests that the polysaccharide GP is a novel antioxidant. ?? 2011 Academic Journals.

  19. Transport across rat trachea in vitro after exposure to cytoskeleton-active drugs in vitro or to ozone in vivo

    SciTech Connect

    Rasmussen, R.E.; Bhalla, D.K.

    1989-03-01

    Full-length tracheas from Sprague-Dawley rats were exposed to cytoskeleton-active drugs in short-term organ culture, and the permeability of the tracheal epithelium was measured by instilling radiotracers into the lumen and assay of the radioactivity appearing in the external bathing medium. In vitro treatment with cytochalasin D (cyto D, 2-10 x 10(-6) M) increased the rate of movement of (14C)mannitol across the epithelium. Exposure to vinblastine (VB, 10(-4) M) alone had no significant effect. However, VB in combination with cyto D increased the permeability in a dose-dependent manner. In vivo exposure to ozone (O3, 0.8 or 2.0 ppm, 2 h) had only a slight effect on the rate of movement of the tracer as measured in vitro immediately after exposure. At 24 h postexposure there was no significant difference in permeability between ozone- and air-exposed tracheas. Prior in vivo O3 exposure sensitized the tracheas to the in vitro effects of cyto D; treatment of O3-exposed tracheas with cyto D immediately after O3 exposure produced a greater than additive effect on permeability measured in vitro. VB at concentrations up to 10(-4) M had no enhancing effect on permeability in O3-exposed tracheas. Sham exposure to clean air did not affect permeability compared to untreated (shelf) controls. Electron microscopic studies demonstrated penetration of horseradish peroxidase into intercellular spaces in the tracheas treated in vitro with cyto D or cyto D plus VB. Cyto D is known to affect intracellular microfilaments that have attachments at or near the cell surface, while VB affects microtubules associated with internal cellular structures. Therefore, the synergistic effect on tracheal permeability observed with O3 and cyto D, but not with O3 and VB, suggests that O3 may change cell surface structures associated with the microfilamentous cytoskeleton.

  20. In vitro accelerated mass propagation and ex vitro evaluation of Aloe vera L. with aloin content and superoxide dismutase activity.

    PubMed

    Gantait, Saikat; Mandal, Nirmal; Das, Prakash Kanti

    2011-08-01

    An innovative protocol on accelerated in vitro propagation and acclimatisation was developed in Aloe vera L. Culture was initiated with rhizomatous stem where Murashige and Skoog (MS) medium fortified with 0.5?mg?L(-1) ?-naphthalene acetic acid and 1.5?mg?L(-1) N(6)-benzylaminopurine (BAP) promoted earliest shoot induction. Maximum shoot multiplication was achieved in MS medium supplemented with 2.5?mg?L(-1)BAP. The best in vitro rooting was observed in the MS medium with 0.5?mg?L(-1) indole-3-acetic acid plus 2?g?L(-1) activated charcoal. The simple acclimatisation process, primarily with a combination of sand and soil (1?:?1 v/v) and finally with a blend of sand, soil and farm yard manure (2?:?1?:?1 v/v), ensured a 98% survival rate. Overall, 192 true-to-type plantlets were achieved from a single explant within 85 days. Morphologically, in vitro generated plants performed better than conventionally propagated plants; nevertheless the similarity in aloin content, gel content and superoxide dismutase activity was corroborated. PMID:21859262

  1. Inhibition of Transcription in Vitro by Anticancer Active Dirhodium(II) Karn Sorasaenee,

    E-print Network

    Turro, Claudia

    Inhibition of Transcription in Vitro by Anticancer Active Dirhodium(II) Complexes Karn SorasaeneeVersity, Columbus, Ohio 43210 Received October 2, 2002 The DNA binding and inhibition of transcription in vitro]/[DNA], of Rh2(µ-O2CCH3)4 and cis-[Rh2(µ-O2CCH3)2(phen)2]2+ at which 50% of the transcription is inhibited (Rinh

  2. Luliconazole Demonstrates Potent In Vitro Activity against Dermatophytes Recovered from Patients with Onychomycosis

    PubMed Central

    Wiederhold, Nathan P.; Fothergill, Annette W.; McCarthy, Dora I.

    2014-01-01

    The in vitro activities of luliconazole, amorolfine, ciclopirox, and terbinafine were determined against 320 dermatophyte isolates from large toenails of onychomycosis patients enrolled into an ongoing phase 2b/3 clinical study. The geometric mean MIC for luliconazole was 0.00022 ?g/ml against all isolates, compared to 0.0194 to 0.3107 ?g/ml for the three other agents. The in vitro potency of luliconazole was maintained regardless of the dermatophyte species. PMID:24709260

  3. The effect of recombinant tissue plasminogen activator on MMP-2 and MMP-9 activities in vitro.

    PubMed

    Golab, Piotr; Kielbus, Michal; Bielewicz, Joanna; Kurzepa, Jacek

    2015-01-01

    One of the most significant side effects during recombinant tissue plasminogen activator (rtPA) for acute stroke treatment is intracranial bleeding. Gelatinases [matrix metalloproteinase (MMP)-2 and MMP-9] are one of the agents involved in the blood-brain barrier destruction resulting in secondary bleeding into the ischemic area during stroke. Previous papers revealed that patients with high baseline MMP-9 serum level have higher risk of intracranial bleeding after thrombolytic therapy. Our objective was to evaluate rtPA influence on serum MMP-2 and MMP-9 activities in vitro. Nine sera obtained from healthy donors were applied for experiment. The commercially available rtPA (Actylise) were diluted with included solvent and additionally with phosphate-buffered saline (PBS) to get concentrations: 2, 4, 8, and 16 ?g/ml. Next, 100 ?l of serum was mixed with equal proportion with different concentrations of rtPA to obtain final rtPA concentrations: 1, 2, 4, and 8 ?g/ml. The sera together with rtPA were incubated for 1 or 2 hours at 37 °C. The activity of gelatinases was estimated with zymography. The activities of MMP-9 (92 kDa) and MMP-2 (72 kDa) were increased by incubation with rtPA in a dose-dependent manner. Simultaneously, the activity of band at 200 kDa (MMP-9/MMP-9 homodimer) was decreased. The activity of gelatinases incubated for 2 hours was elevated in comparison with 1-hour incubation; however, the increase was observed even for sample without rtPA. In conclusion, this study showed that rtPA can increase the biological activity of MMP-2 and MMP-9 on posttranslational level. PMID:24963695

  4. In vitro antioxidant activity of Holarrhena antidysenterica Wall. methanolic leaf extract

    PubMed Central

    Ganapathy, P. S. Sujan; Ramachandra, Y. L.; Rai, S. Padmalatha

    2011-01-01

    Antioxidative potential of methanolic leaf extract of Holarrhena antidysenterica was evaluated using hydroxyl radical, superoxide anion scavenging and reducing power assays. The antioxidant activity of the methanol extract increased in a concentration-dependent manner. The extract showed significant reactive oxygen species (ROS) scavenging activity in all in vitro antioxidant assays and contained high level of total phenolic content PMID:24826020

  5. Activation of microglia by Borna disease virus infection: in vitro study

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Neonatal Borna disease virus (BDV) infection of the rat brain is associated with microglial activation and damage to the certain neuronal populations. Since persistent BDV infection of neurons in vitro is noncytolytic and noncytopathic, activated microglia have been suggested to be responsible for n...

  6. In vitro biological activity of decoction of Joshanda.

    PubMed

    Abdullah; Inayat, Humaira; Khan, Haroon; Khan, Lajber; Khan, Mohammad Iqbal; Hassan, Sohail; Khan, Murad Ali

    2014-03-01

    Joshanda is a polyherbal product, commonly practicing in inflammation of upper respiratory tract as tea. The present study was conducted to find out its antimicrobial, phytotoxic, leishmanicidal and cytotoxic activities. The decoction of the product showed profound activity against Gram positive tested pathogens especially S. aureus 36.5 mm zone of inhibition at 8.0 ?g/ml. However, it was inactive against C. albicans. Closed correlation was observed between two methods in terms of results. It had potent phytotoxic activity (75%). However, it was devoid of any activity leishmanicidal and cytotoxic activity. Phytochemical studies of Joshanda showed the presence of various pharmacologically active groups. PMID:24577908

  7. In vitro antifungal activity of pneumocandin L-743,872 against a variety of clinically important molds.

    PubMed Central

    Del Poeta, M; Schell, W A; Perfect, J R

    1997-01-01

    The in vitro activity of the new antifungal drug pneumocandin L-743,872 against 55 isolates of clinically important molds was examined by an adapted macrobroth dilution method for yeasts. Pneumocandin L-743,872 exhibited in vitro antifungal activity against Alternaria sp., Aspergillus flavus, Aspergillus fumigatus, Curvularia lunata, Exophiala jeanselmei, Fonsecaea pedrosoi, Paecilomyces variotii, and Scedosporium apiospermum. The drug appeared to lack significant in vitro inhibitory activity against Fusarium oxysporum, Fusarium solani, Rhizopus arrhizus, Paecilomyces lilacinus, and Scedosporium prolificans. PMID:9257774

  8. Bromelain enzyme from pineapple: in vitro activity study under different micropropagation conditions.

    PubMed

    Vilanova Neta, Jaci Lima; da Silva Lédo, Ana; Lima, Aloisio André Bonfim; Santana, José Carlos Curvelo; Leite, Nadjma Souza; Ruzene, Denise Santos; Silva, Daniel Pereira; de Souza, Roberto Rodrigues

    2012-09-01

    The aim of this work was to evaluate the activity of bromelain in pineapple plants (Ananas comosus var. Comosus), Pérola cultivar, produced in vitro in different culture conditions. This enzyme, besides its pharmacological effects, is also employed in food industries, such as breweries and meat processing. In this work, the enzymatic activity was evaluated in the tissues of leaves and stems of plants grown in culture medium without plant growth regulator. The most significant levels of bromelain were observed in leaf tissue after 4 months of culture in vitro in medium with a filter paper bridge, followed by medium gelled by the agar. The results of this study, regarding the different structures of the pineapple (leaves and stems) in vitro showed that the activity of bromelain varied depending on the culture conditions, the time and structure of which was quantified, ensuring a viable strategy in the production of seedlings with high levels of bromelain in subsequent phases of micropropagation. PMID:22736274

  9. In vitro and in vivo biological activities of anthocyanins from Nitraria tangutorun Bobr. fruits.

    PubMed

    Ma, Tao; Hu, Na; Ding, Chenxi; Zhang, Qiulong; Li, Wencong; Suo, Yourui; Wang, Honglun; Bai, Bo; Ding, Chenxu

    2016-03-01

    Anthocyanins are the main compounds in Nitraria tangutorun Bobr. The enrichment and purification of anthocyanins on macroporous resins were investigated. Regarding anthocyanin purification, static adsorption and desorption were studied. The optimal experimental conditions were the following: resin type: X-5; static adsorption time: 6h; desorption solution: ethanol-water-HCl (80:19:1, V/V/V; pH 1); desorption time: 40min. Furthermore, the in vitro and in vivo biological activities of the anthocyanins were evaluated. The anthocyanins showed ideal scavenging effects on free radicals in vitro, especially on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl free radical (OH). In the animal experiment, blood lipid metabolism of hyperlipidemia rats was regulated by anthocyanin contents. The superoxide dismutase (SOD) activity and the total antioxidant capacity (TAC) of hyperlipidemia rats were also improved by anthocyanins. These results showed that anthocyanins from N. tangutorun Bobr. fruits had potential biological activities in vivo as well as in vitro. PMID:26471558

  10. Anticoagulant activities of piperlonguminine in vitro and in vivo

    PubMed Central

    Lee, Wonhwa; Yoo, Hayoung; Ku, Sae-Kwang; Kim, Jeong Ah; Bae, Jong-Sup

    2013-01-01

    Piperlonguminine (PL), an important component of Piper longum fruits, is known to exhibit anti-hyperlipidemic, antiplatelet and anti-melanogenic activities. Here, the anticoagulant activities of PL were examined by monitoring activatedpartial-thromboplastin-time (aPTT), prothrombin-time (PT), and the activities of thrombin and activated factor X (FXa). The effects of PL on the expressions of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were also tested in tumor necrosis factor-? (TNF-?) activated HUVECs. The results showed that PL prolonged aPTT and PT significantly and inhibited the activities of thrombin and FXa. PL inhibited the generation of thrombin and FXa in HUVECs. In accordance with these anticoagulant activities, PL prolonged in vivo bleeding time and inhibited TNF-? induced PAI-1 production. Furthermore, PAI-1/t-PA ratio was significantly decreased by PL. Collectively, our results suggest that PL possesses antithrombotic activities and that the current study could provide bases for the development of new anticoagulant agents. [BMB Reports 2013; 46(10): 484-489] PMID:24148768

  11. In-vitro anticoagulant activity of fucoidan derivatives from brown seaweed Laminaria japonica

    NASA Astrophysics Data System (ADS)

    Wang, Jing; Zhang, Quanbin; Zhang, Zhongshan; Hou, Yun; Zhang, Hong

    2011-05-01

    Fucoidan, a group of sulfated heteropolysaccharides, was extracted from Laminaria japonica, an important economic alga species in China. The anticoagulant activity of fucoidan and its derivatives (including sulfated, phosphorylated, and aminated fucoidan) was examined using in-vitro anticoagulant systems. The correlation between chemical variations within the fucoidan group and anticoagulant activity was determined. The in-vitro anticoagulant properties of fucoidan and its derivatives were determined by measuring activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). The results indicate anticoagulant activity in all samples using APTT and TT assays; however, only the fucoidan derivatives affected the PT assay. Thus, the fucoidan derivatives were able to inhibit both intrinsic and extrinsic blood coagulants. Fucoidan (FPS) and its derivatives presented better anticoagulant activity than low molecular weight fucoidan (DFPS) and its derivatives, suggesting that molecular weight and proper conformation are contributing factors for anticoagulant activity of polysaccharides. Amino groups have a positive charge and can thus change the charge density of fucoidan. Accordingly, among the tested samples, aminated fucoidan (NF) was the most active reflecting the importance of charge density for anticoagulant activity. Available data obtained using in-vitro models suggest that the sulfate content, sulfate/total-sugar ratio, molecular weight, and the substituted group of fucoidan are important factors for anticoagulant activity but that the influence of sulfate, phosphate and amino groups on anticoagulant activity was different.

  12. In vitro and in vivo activities of Peganum harmala extract against Leishmania major

    PubMed Central

    Rahimi-Moghaddam, Parvaneh; Ebrahimi, Soltan Ahmed; Ourmazdi, Hourmazd; Selseleh, Monawar; Karjalian, Maryam; Haj-Hassani, Giti; Alimohammadian, Mohammad Hossein; Mahmoudian, Massoud; Shafiei, Massoumeh

    2011-01-01

    BACKGROUND: In vitro and in vivo antileishmanial activities of crude hydroalcoholic extract of peganum harmala seeds were investigated against Leishmania major. METHODS: The extract of aerial parts of P harmala was obtained by maceration. The in vitro experiments were performed on promastigotes to assess antileishmanial activity of the extract using amphotericin B as a reference. The in vivo studies were carried out on cutaneous leishmaniasis in outbred mice to evaluate the effects of topical application of the ointment-based extract. RESULTS: The in vitro experiments showed a concentration-dependent decrease of parasites number caused by the extract with an IC50 value of 59.4 ?g/ml. In vivo studies demonstrated a significant post-treatment decrease in the lesion size and parasite count in infected animals, compared to placebo and control groups. High performance liquid chromatography (HPLC) of the crude extract demonstrated the existence of harmaline and harmine as beta-carboline alkaloids. CONCLUSIONS: P harmala seeds extract showed significant in vitro and in vivo antileishmanial activities. Most biological activity of the extract could be attributed to its beta-carboline content. However, another alkaloid of P harmala seeds extract, peganine, has also been reported to have antileishmanial activity. These beneficial effects can be attributed to the cumulative effects of various biologically active components present in it. PMID:22279479

  13. Pulsatile activity and hatching of in vitro produced cow blastocysts: effects of serum supplementation.

    PubMed

    van Heule, A; van Langendonckt, A; Donnay, I; Dessy, F; Massip, A

    2001-04-01

    The effect of serum added to a modified SOF medium on pulsatile activity and hatching of in vitro produced cow blastocysts was investigated by time-lapse cinematography. Embryos were generated from abattoir material and cultured in mSOF without serum or with 10% FCS added at 42h pi. Addition of serum significantly increases pulsatile activity before zona rupture and reduces the time of hatching. Pulsatile activity does not seem to be involved in the hatching process. PMID:11778742

  14. Absence of in vitro Procoagulant Activity in Immunoglobulin Preparations due to Activated Coagulation Factors

    PubMed Central

    Oviedo, Adriana E.; Bernardi, María E.; Guglielmone, Hugo A.; Vitali, María S.

    2015-01-01

    Summary Background Immunoglobulin (IG) products, including intravenous (IVIG) or subcutaneous (SCIG) immunoglobulins are considered safe and effective for medical therapy; however, a sudden and unexpected increase in thromboembolic events (TE) after administration of certain batches of IVIG products has been attributed to the presence of activated coagulation factors, mainly factor XIa. Our aims were to examine the presence of enduring procoagulant activity during the manufacturing process of IGs, with special focus on monitoring factor XIa, and to evaluate the presence of in vitro procoagulant activity attributed to coagulation factors in different lots of IVIG and SCIG. Methods Samples of different steps of IG purification, 19 lots of IVIG and 9 of SCIG were analyzed and compared with 1 commercial preparation of IVIG and 2 of SCIG, respectively. Factors II, VII, IX, XI and XIa and non-activated partial thromboplastin time (NAPTT) were assayed. Results The levels of factors II, VII, IX, X and XI were non-quantifiable once fraction II had been re-dissolved and in all analyzed lots of IVIG and SCIG. The level of factor XIa at that point was under the detection limits of the assay, and NAPTT yielded values greater than the control during the purification process. In SCIG, we detected higher concentrations of factor XIa in the commercial products, which reached values up to 5 times higher than the average amounts found in the 9 batches produced by UNC-Hemoderivados. Factor XIa in commercial IVIG reached levels slightly higher than those of the 19 batches produced by UNC-Hemoderivados. Conclusion IVIG and SCIG manufactured by UNC-Hemoderivados showed a lack of thrombogenic potential, as demonstrated not only by the laboratory data obtained in this study but also by the absence of any reports of TE registered by the post marketing pharmacovigilance department.

  15. Novel trisubstituted harmine derivatives with original in vitro anticancer activity.

    PubMed

    Frédérick, Raphaël; Bruyčre, Céline; Vancraeynest, Christelle; Reniers, Jérémy; Meinguet, Céline; Pochet, Lionel; Backlund, Anders; Masereel, Bernard; Kiss, Robert; Wouters, Johan

    2012-07-26

    To overcome the intrinsic resistance of cancer cells to apoptotic stimuli, we designed and synthesized approximately 50 novel ?-carbolines structurally related to harmine. Harmine is known for its anticancer properties and is a DYRK1A inhibitor. Of the synthesized compounds, the most active in terms of growth inhibition of five cancer cell lines are cytostatic and approximately 100 times more potent than harmine but demonstrated no DYRK1A inhibitory activity. These novel ?-carbolines display similar growth inhibitory activity in cancer cells that are sensitive and resistant to apoptotic stimuli. Using ChemGPS-NP, we found that the more active ?-carbolines are all more lipophilic and larger than the less active compounds. Lastly, on the basis of the NCI human tumor cell line anticancer drug screen and the NCI COMPARE algorithm, it appears that some of these compounds, including 5a and 5k, seem to act as protein synthesis inhibitors. PMID:22770529

  16. Antiviral activity of doxycycline against vesicular stomatitis virus in vitro.

    PubMed

    Wu, Zhuan-Chang; Wang, Xin; Wei, Jian-Chao; Li, Bei-Bei; Shao, Dong-Hua; Li, Yu-Ming; Liu, Ke; Shi, Yuan-Yuan; Zhou, Bin; Qiu, Ya-Feng; Ma, Zhi-Yong

    2015-11-01

    Doxycycline (Dox) is a tetracycline derivative with broad-spectrum antimicrobial activities that is used as an effector substance in inducible gene-expression systems. We investigated the antiviral activity of Dox against vesicular stomatitis virus (VSV) infection in cultured H1299 cells. Dox at concentrations of 1.0-2.0 ?g ml(-1) significantly inhibited VSV replication and the VSV-induced cytopathic effect in dose-dependent manners, suggesting that Dox may have broader activity in inhibiting viral replication, in addition to its well-defined bacteriostatic activity. Dox exerted its antiviral effect at the early-mid stage of VSV infection, suggesting that it did not interfere with VSV infectivity, adsorption, or entry into target cells. These results indicate that Dox can inhibit VSV infection and may therefore have potential applications for the treatment of viral infections. PMID:26459887

  17. Impact of antibacterial drugs on human serum paraoxonase-1 (hPON1) activity: an in vitro study

    PubMed Central

    Söyüt, Hakan; Kaya, Elif Duygu; Beydemir, ?ükrü

    2014-01-01

    Objective To investigate the in vitro effects of the antibacterial drugs, meropenem trihydrate, piperacillin sodium, and cefoperazone sodium, on the activity of human serum paraoxonase (hPON1). Methods hPON1 was purified from human serum using simple chromatographic methods, including DEAE-Sephadex anion exchange and Sephadex G-200 gel filtration chromatography. Results The three antibacterial drugs decreased in vitro hPON1 activity. Inhibition mechanisms meropenem trihydrate was noncompetitive while piperacillin sodium and cefoperazone sodium were competitive. Conclusions Our results showed that antibacterial drugs significantly inhibit hPON1 activity, both in vitro, with rank order meropenem trihydrate piperacillin sodium cefoperazone sodium in vitro. PMID:25183328

  18. In vitro determination of the spermicidal activity of plant saponins.

    PubMed

    Primorac, M; Sekulovi?, D; Antoni?, S

    1985-08-01

    The plant kingdom might yield an effective antifertility drug. A Mentha arvensis L. (Labiatae) fraction with uterotonic activity was isolated, and was found to be active on the nonpregnant as well as the pregnant rat uterus. According to folklore medicine, the Mexican plant Montanoa tomentosa Cerv. (zoapatle) possesses antifertility activity in women. The effect of various isolated preparations from this plant on early pregnancy were investigated in serveral rodent species including the mouse, rat hamster, and guinea pig. It was concluded that zoapatle plant extracts possess unique antifertility activity. Lin-Hsim and coworkers isolated fractions from Aristolochia molissima Hanceith contrceptive activity in female mice. Saponins of some plants were used in contraceptive formulations either as foaming agents or as spermicidal substances. Elbary and Nour investigated the spermicidal effects of saponins isolated from the following plants: Gypsophila paniculata L., Saponaria officinalis L., Enterolobium cyclocarpum, Griseb., Terminalia horrida Steud., Melilotus sicula Vitm., and Ruscus hypoglossum L. All of the saponins tested possessed spermicidal activity. Jain and coworkers isolated 2 new saponins in Pittosporum nilghrense with spermicidal effects. In this paper we have determined the spermicidal activity of saponins isolated from some Yugoslav plants, which in that aspect have not been investigated. The results are illustrated in the table. They show that all of the saponins tested were spermicidal in dependence on their nature. Saponins of Primula vulgaris Huds. and Cyclamen persicum Mill. immobilized human spermatozoa within a period of 20 s at a dilution 1:1000. Saponin of Gypsophila paniculata L. was spermicidal at dilution 1:20. These findings show that saponins isolated from some Yugoslav plants may be useful spermicides of natural origin. PMID:4080814

  19. In vitro activities of four xyloglucan endotransglycosylases from Arabidopsis

    NASA Technical Reports Server (NTRS)

    Campbell, P.; Braam, J.; McIntire, L. V. (Principal Investigator)

    1999-01-01

    Xyloglucan endotransglycosylases (XETs) are encoded by a gene family in Arabidopsis thaliana. These enzymes modify a major structural component of the plant cell wall, xyloglucan, and therefore may influence plant growth and development. We have produced four Arabidopsis XETs (TCH4, Meri-5, EXGT and XTR9) using the baculovirus/insect cell system and compared their biochemical activities. TCH4, as previously demonstrated, and the other three proteins are capable of carrying out transglycosylation of xyloglucans. The K(m) for XLLGol acceptor oligosaccharide is in the range of 20-40 microM for all the XETs except XTR9, which has a Km of 5 microM and is significantly inhibited by high levels of XLLGol. All four enzymes are most active between pH 6.0 and 6.5. TCH4 and XTR9 have temperature optima of 18 degrees C, whereas Meri-5 and EXGT are most active at 28 and 37 degrees C, respectively. Although the activity levels of three of the XETs are not influenced by the presence of fucose on the xyloglucan polymer, XTR9 has a clear preference for non-fucosylated xyloglucan polymer. The four XETs show a marked preference for XLLGol over either XXFGol or XXXGol as acceptor oligosaccharide. All four XETs are glycosylated; however, only the activities of TCH4 and Meri-5 are affected by the removal of the N-glycan with PNGase F. These four enzymes most likely function solely as transglycosylases because xyloglucan endoglucanase activity was not apparent. Subtle differences in biochemical activities may influence the physiological functions of the distinct XETs in vivo.

  20. In vitro activity of Amazon plant extracts against Enterococcus faecalis

    PubMed Central

    de Castilho, Adriana Lígia; da Silva, Juliana Paola Correa; Saraceni, Cintia Helena Coury; Díaz, Ingrit Elida Collantes; Paciencia, Mateus Luís Barradas; Varella, Antonio Drauzio; Suffredini, Ivana Barbosa

    2014-01-01

    Previous studies analyzing 2,200 plant extracts indicated anti-enterococcal activity in 25 extracts obtained from Brazilian forests’ plants. In the present study, these extracts were subjected to microdilution broth assay (MDBA) and disk diffusion assay (DDA) using planktonic Enterococcus faecalis ATCC® 29212™ and were submitted to phytochemical analysis in TLC and HPLC. Three extracts obtained from Ipomoea alba (MIC < 40 ?g/mL), Diclinanona calycina (MIC ? 40 ?g/mL) and Moronobea coccinea (40 < MIC < 80 ?g/mL; MBC = 80 ?g/mL) showed significant bactericidal activity in the MDBA and four extracts obtained from I. alba (14.04 ± 0.55 mm diameter) S. globulifera (14.43 ± 0.33 mm and 12.18 ± 0.28 mm diameter) and Connarus ruber var. ruber (13.13 ± 0.18 mm diameter) were active in DDA. Residues H2O obtained from Psidium densicomum (mean of 16.78 mm diameter) and from Stryphnodendron pulcherrimum (mean of 15.97 mm diameter) have shown an improved antibacterial activity after fractionation if compared to that obtained from the respective crude extracts. Antioxidant activity was observed in some residues of the active extracts. TLC analysis showed that phenolic compounds are likely to be found in active extracts. Three molecules were isolated from S. globulifera and were identified by 13C NMR lupeol, ?-amyrin and 3?-hydroxyglutin-5-ene. The present chemical and biological findings suggest that these extracts are a potential source of new anti-Enterococcus compounds to be introduced in endodontic therapy. PMID:25477906

  1. In vitro antibacterial activity of different pulp capping materials

    PubMed Central

    Beltrami, Riccardo; Colombo, Marco; Ceci, Matteo; Dagna, Alberto; Chiesa, Marco

    2015-01-01

    Background Direct pulp capping involves the application of a dental material to seal communications between the exposed pulp and the oral cavity (mechanical and carious pulp exposures) in an attempt to act as a barrier, protect the dental pulp complex and preserve its vitality. The aim of this study was to evaluate and compare, by the agar disc diffusion test, the antimicrobial activity of six different pulp-capping materials: Dycal (Dentsply), Calcicur (Voco), Calcimol LC (Voco), TheraCal LC (Bisco), MTA Angelus (Angelus), Biodentine (Septodont). Material and Methods Streptococcus salivarius, Streptococcus sanguis and Streptococcus mutans strains were selected to evaluate the antimicrobial activity by the agar disc diffusion test of different pulp capping materials. Paper disks were impregnated whit each pulp capping materials and placed onto culture agar-plates pre-adsorbed with bacterial cells and further incubated for 24 h at 37°C. The growth inhibition zones around each pulp capping materials were recorded and compared for each bacterial strain. Results For the investigation of the antibacterial properties the ANOVA showed the presence of significant differences among the various materials. Tukey test showed that MTA-based materials induced lower growth inhibition zones. Conclusions MTA-based products show a discrete antibacterial activity varying from calcium hydroxide-based materials which present an higher antibacterial activity. Key words:Agar disc diffusion test, antimicrobial activity, calcium hydroxide, MTA, pulp capping materials. PMID:26644833

  2. In vivo expression of in vitro anticoccidial activity.

    PubMed

    Ricketts, A P; Olson, J A; Rice, J R

    1992-10-01

    Large-scale screening has led to the identification of several experimental compounds that have very potent intrinsic activity against coccidia, but the lack of translation to in vivo efficacy has been a major hurdle in developing such leads into effective new drugs. We developed methods to explore the impact of oral availability and appropriate distribution in tissue, both of which are potentially important factors in the expression of activity in vivo. For the compounds that we examined, neither oral absorption nor distribution to the site of infection appeared to be the critical barrier to in vivo expression of intrinsic anticoccidial activity. Elucidation of the nature of additional factors that might be involved could assist greatly in the identification of useful new anticoccidial agents. PMID:1444315

  3. In vivo expression of in vitro anticoccidial activity.

    PubMed Central

    Ricketts, A P; Olson, J A; Rice, J R

    1992-01-01

    Large-scale screening has led to the identification of several experimental compounds that have very potent intrinsic activity against coccidia, but the lack of translation to in vivo efficacy has been a major hurdle in developing such leads into effective new drugs. We developed methods to explore the impact of oral availability and appropriate distribution in tissue, both of which are potentially important factors in the expression of activity in vivo. For the compounds that we examined, neither oral absorption nor distribution to the site of infection appeared to be the critical barrier to in vivo expression of intrinsic anticoccidial activity. Elucidation of the nature of additional factors that might be involved could assist greatly in the identification of useful new anticoccidial agents. PMID:1444315

  4. Alternatively activated dendritic cells regulate CD4+ T-cell polarization in vitro and in vivo

    E-print Network

    Allen, Judith

    Peter C. Cooka,1 , Lucy H. Jonesa,1 , Stephen J. Jenkinsa , Thomas A. Wynnb , Judith E. Allena and cumulative function of these IL-4­ induced products is not clear, aaMs are thought to play vital rolesAlternatively activated dendritic cells regulate CD4+ T-cell polarization in vitro and in vivo

  5. Lasting modulation of in-vitro oscillatory activity with weak direct current stimulation

    E-print Network

    Parra, Lucas C.

    University of New York, New York, USA December 3, 2014 Abstract Transcranial Direct Current Stimulation (t hypothesis. Introduction The number of studies on transcranial direct current stimulation (tDCS) has rapidlyLasting modulation of in-vitro oscillatory activity with weak direct current stimulation Davide

  6. Porcine Splenic Hydrolysate has Antioxidant Activity in vivo and in vitro

    PubMed Central

    Yoon, Taek Joon

    2014-01-01

    The antioxidant capacity of porcine splenic hydrolysate (PSH) was studied in vitro and in vivo. Peptide hydrolysates were prepared, using the proteolytic enzyme Alcalase®. The molecular weights of PSH were 37,666, 10,673, 6,029, and 2,918 g/mol. Rats were fed a 5% (w/v) PSH diet, instead of a casein diet, for 4 wk. The food intake, body weight gain, and liver weight of rats in the PSH group were similar to those in the control (CONT) group. There were no differences in the serum total cholesterol, triglyceride, total protein, or albumin levels between PSH and CONT groups. However, the level of in vivo hepatic lipid peroxidation in PSH group was significantly lower than that in CONT. In vivo hepatic catalase and glutathione peroxidase activities in the PSH group were significantly higher than those in the control group. The in vitro protein digestibility of PSH was lower than that of casein. The in vitro trolox equivalent antioxidant capacity of PSH was significantly higher than that of the peptide hydrolysate from casein. The in vitro radical scavenging activities of PSH were significantly higher than those of the peptide hydrolysate from casein. The present findings suggest that porcine splenic peptides improve the antioxidant status in rats by enhancing hepatic catalase and GSH-Px activities, and indicate a potential mechanism of radical scavenging activity during gastrointestinal passage.

  7. IN VITRO ANDROGENIC ACTIVITY OF KRAFT MILL EFFLUENT IS ASSOCIATED WITH MASCULINIZATION OF FEMALE FISH

    EPA Science Inventory

    In Vitro Androgenic Activity of Kraft Mill Effluent is Associated with Masculinization of Female Fish. Lambright, CS 1 , Parks, LG 1, Orlando, E 2, Guillette, LJ, Jr.2, Ankley, G 3, Gray, LE, Jr.1 , 1USEPA, NHEERL, RTP, NC, 2 University of Florida, Dept. of Zoology, Gainesville ...

  8. In Vitro and In Vivo Cytotoxicities and Antileishmanial Activities of Thymol and Hemisynthetic Derivatives

    PubMed Central

    Robledo, Sara; Osorio, Edison; Muńoz, Diana; Jaramillo, Luz Marina; Restrepo, Adriana; Arango, Gabriel; Vélez, Iván

    2005-01-01

    The in vitro and in vivo antileishmanial and cytotoxic activities of thymol and structural derivatives in comparison to those of Glucantime were studied. The results showed here suggest that thymol and hemisynthetic derivatives have promising antileishmanial potential and could be considered as new lead structures in the search for novel antileishmanial drugs. PMID:15793164

  9. “In vitro” antifungal activity of ozonized sunflower oil on yeasts from onychomycosis

    PubMed Central

    Guerrer, L.V.; Cunha, K. C.; Nogueira, M. C. L.; Cardoso, C. C.; Soares, M. M. C. N.; Almeida, M. T. G.

    2012-01-01

    The “in vitro” antifungal activity of ozonized sunflower oil (Bioperoxoil®) was tested on 101 samples of yeasts originating from onychomycosis using the disk diffusion method. The oil was efficacious against several clinical fungal strains: Candida parapsilosis, Candida albicans, Trichosporon asahii, Candida tropicalis and Candida guilliermondii. PMID:24031958

  10. EFFECT OF WATER POLLUTANTS AND OTHER CHEMICALS UPON THE ACTIVITY OF LIPASE 'IN VITRO'

    EPA Science Inventory

    Lipase preparations were treated in vitro with 100 chemicals of various classes, many of which are environmental pollutants, to determine their effect upon enzyme activity. The greatest inhibition was caused by mercuric ion and certain heavy metal cations; almost as inhibiting we...

  11. In Vitro Activities of Eight Antifungal Drugs against 104 Environmental and Clinical Isolates of Aureobasidium pullulans

    PubMed Central

    Najafzadeh, M. Javad; Sutton, Deanna A.; Keisari, M. Saradeghi; Zarrinfar, H.; de Hoog, G. Sybren; Chowdhary, Anuradha

    2014-01-01

    Aureobasidium pullulans is an unusual agent of phaeohyphomycosis. The in vitro activities of antifungals against 104 isolates of Aureobasidium pullulans var. pullulans and A. pullulans var. melanigenum revealed low MIC90s of amphotericin B, posaconazole, and itraconazole. However, they were resistant to fluconazole (?64 ?g/ml) and had high MICs of voriconazole, isavuconazole, caspofungin, and micafungin. PMID:25001309

  12. In vitro metabolism and bioavailability tests for the predictive toxicology of endocrine active substances

    EPA Science Inventory

    Legislation and prospective legislative proposals internationally (may) require that chemicals are tested for their ability to disrupt the hormonal systems of animals. Chemicals found to test positive in vitro are considered to be endocrine active substances (EAS) and may be puta...

  13. INFLUENCE OF CCL4 BIOTRANSFORMATION ON THE ACTIVATION OF RAT LIVER PHOSPHOLIPASE C IN VITRO

    EPA Science Inventory

    The Influence of CCl4 Biotransforrnation on the Activation of Rat Liver Phospholipase C in Vitro. Coleman, J.F., Condie, L.W. AND LAMB, R.G. (1988). Toxicol. Appl Pharmacol. 95, 200-207. Carbon tetrachloride (CCL4) biotransformation and covalent binding was measured in l000g live...

  14. In Vitro Anthelmintic Activity of Baliospermum montanum Muell. Arg roots.

    PubMed

    Mali, R G; Wadekar, R R

    2008-01-01

    Alcohol and aqueous extracts from the roots of Baliospermum montanum Muell. Arg were investigated for their anthelmintic activity against Pheretima posthuma and Ascardia galli. Various concentrations (10-100 mg/ml) of each extract were tested in the bioassay, which involved determination of time of paralysis and time of death of the worms. Both the extracts exhibited significant anthelmintic activity at highest concentration of 100 mg/ml. Piperazine citrate (10 mg/ml) was included as standard reference and distilled water as control. PMID:20390101

  15. In Vitro Anthelmintic Activity of Baliospermum montanum Muell. Arg roots

    PubMed Central

    Mali, R. G.; Wadekar, R. R.

    2008-01-01

    Alcohol and aqueous extracts from the roots of Baliospermum montanum Muell. Arg were investigated for their anthelmintic activity against Pheretima posthuma and Ascardia galli. Various concentrations (10-100 mg/ml) of each extract were tested in the bioassay, which involved determination of time of paralysis and time of death of the worms. Both the extracts exhibited significant anthelmintic activity at highest concentration of 100 mg/ml. Piperazine citrate (10 mg/ml) was included as standard reference and distilled water as control. PMID:20390101

  16. In vivo and in vitro antileishmanial activity of Bungarus caeruleus snake venom through alteration of immunomodulatory activity.

    PubMed

    Bhattacharya, Shamik; Ghosh, Prasanta; De, Tripti; Gomes, Antony; Gomes, Aparna; Dungdung, Sandhya Rekha

    2013-09-01

    Leishmaniasis threatens more than 350 million people worldwide specially in tropical and subtropical region. Antileishmanial drugs that are currently available have various limitations. The search of new drugs from natural products (plants, animals) possessing antileishmanial activity is ventured throughout the world. The present study deals with the antileishmanial activity of Bungarus caeruleus snake venom (BCV) on in vitro promastigotes and amastigotes of Leishmania donovani parasite and leishmania infected BALB/c mice. The effect of BCV on peritoneal macrophage, release of cytokines from the activated macrophages, production of nitric oxide, reactive oxygen species and cytokines were studied in vivo and in vitro. IC50 value of BCV on L. donovani promastigote was 14.5 ?g/ml and intracellular amastigote was 11.2 ?g/ml. It activated peritoneal macrophages, significantly increased cytokines and interleukin production. BCV (20 ?g/kg and 40 ?g/kg body weight of mice) decreased parasite count by 54.9% and 74.2% in spleen and 41.4% and 60.4% in liver of infected BALB/c mice. BCV treatment significantly increased production of TNF-?, IFN-?, ROS, NO in infected mice. Histological studies showed decreased granuloma formation in treated liver as compared with control. Liver and spleen structure was partially restored due to BCV treatment in infected mice. The present study revealed that BCV possessed antileishmanial activity against L. donovani parasite in vivo and in vitro and this activity was partly mediated through immunomodulatory activity involving macrophages. PMID:23830987

  17. Inhibition of catalase activity in vitro by diesel exhaust particles

    SciTech Connect

    Mori, Yoki; Murakami, Sumika; Sagae, Toshiyuki

    1996-02-09

    The effect of diesel exhaust particles (DEP) on the activity of catalase, an intracellular anti-oxidant, was investigated because H{sub 2}O{sub 2} is a cytotoxic oxidant, and catalase released from alveolar cells is an important antioxidant in the epithelial lining fluid in the lung. DEP inhibited the activity of bovine liver catalase dose-dependently, to 25-30% of its original value. The inhibition of catalase by DEP was observed only in the presence of anions such as Cl{sup {minus}}, Br{sup {minus}}, or thiocyanate. Other anions, such as CH{sub 3}COO{sup {minus}} or SO{sub 4}{sup {minus}}, and cations such as K{sup +}, Na{sup +}, Mg{sup 2+}, or Fe{sup 2+}, did not affect the activity of catalase, even in the presence of DEP extract. Catalase from guinea pig alveolar cells and catalase from red blood cells were also inhibited by DEP extracts, as was catalase from bovine liver. These results suggest that DEP taken up in the lung and located on alveolar spaces might cause cell injury by inhibiting the activity of catalase in epithelial lining fluid, enhancing the toxicity of H{sub 2}O{sub 2} generated from cells in addition to that of O{sub 2}{sup {minus}} generated by the chemical reaction of DEP with oxygen. 10 refs., 6 figs.

  18. In vitro antioxidant and antitumor activities of different sulfated polysaccharides isolated from three algae.

    PubMed

    Shao, Ping; Chen, Xiaoxiao; Sun, Peilong

    2013-11-01

    Three sulfated polysaccharides(Ulva fasciata (UFP), Gloiopeltis furcata (GFP), Sargassum henslouianum (SHP))were isolated from three algae including green alga Ulva fasciata, red alga Gloiopeltis furcata and brown alga Sargassum henslouianum by ultrasonic extraction and radial flow chromatography. Their in vitro antioxidant and antitumor activities were investigated and compared. Among these three polysaccharides, UFP, with relatively lower sulfate content, exhibited excellent antioxidant activities in superoxide radical assay, ABTS assay and DPPH assay; however, it demonstrated the minimal inhibitory effects on growth of MKN45 gastric cancer cells and DLD intestinal cancer cells. SHP with the lowest sulfate content gained relatively lower radical scavenging rates but showed significantly higher antitumor activities. These results indicated that the in vitro antitumor and antioxidant activities of the three polysaccharides may be related to combined effects of sulfate content and uronic acid content. PMID:23994786

  19. Effect of Low Dose Gamma Irradiation together with Lipid A on Human Leukocytes Activities In Vitro

    NASA Astrophysics Data System (ADS)

    Belyakova, E.; Dubnickova, M.; Boreyko, A.

    2010-01-01

    The influence of gamma irradiation and of Lipid A from Escherichia coli on phagocytosis, lyzosyme and peroxidase activities of human leukocytes, in vitro was investigated. Leukocytes samples were irradiated with 1 and 5 Gy, respectively. The number of irradiated leukocytes was decreased in the irradiated samples. Only samples with additive Lipid A were not damaged by irradiation. The Lipid A had positive influence on biological activities of the irradiated leukocytes.

  20. In vitro antibacterial activity of concentrated polyethylene glycol 400 solutions.

    PubMed Central

    Chirife, J; Herszage, L; Joseph, A; Bozzini, J P; Leardini, N; Kohn, E S

    1983-01-01

    It was found that concentrated polyethylene glycol 400 (PEG 400) solutions have significant antibacterial activity against various pathogenic bacteria, including Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus. This effect might be attributed to two effects: lowering of water activity and, superimposed on this, the specific action of PEG-400 molecules on bacterial cells. Phase-contrast microscopic observations of cells placed in contact with PEG 400 revealed clumping and morphological changes of bacterial cells. The larger changes in appearance were evidenced by the species which were more rapidly killed by PEG 400. The results obtained suggested that concentrated PEG 400 solutions may have a potential value in medicine as a topical antibacterial agent. The feasibility of this application is the subject of present investigation. Images PMID:6638996

  1. In Vitro Larvicidal and Antioxidant Activity of Dihydrophenanthroline-3-carbonitriles

    PubMed Central

    Bharathi, A.; Rahuman, Abdul Abdul; Rajakumar, Govindasamy

    2014-01-01

    Many naturally occurring and synthetic compounds containing dihydrocyanopyridine and cyanopyran moiety show pharmacological properties. The aim of this study is to investigate the larvicidal and antioxidant potential of dihydrophenanthroline-3-carbonitrile derivatives 4a–f. A novel series of 2-amino-10-chloro-4,12-diphenyl-1,4,5,6-tetrahydrobenzo[j][1,7]phenanthroline-3-carbonitrile derivatives were synthesized by reacting different substituted acridine chalcones through Michel addition. The compounds were synthesized in excellent yields and the structures were corroborated on the basis of FT-IR, 1H NMR, 13C NMR, and ESI Mass analysis data. All the synthesized compounds were evaluated for larvicidal activity against Aedes aegypti and Culex quinquefasciatus larvae. Furthermore, the antioxidant activity was studied by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay method. From the antioxidant assay, the compound 4c was reported with profound antioxidant potential. PMID:24868553

  2. In vitro Evaluation of Anthelmintic Activity of Nauclea orientalis Leaves.

    PubMed

    Raghavamma, S T V; Rao, N Rama

    2010-07-01

    Antianthelmintic activity of successive extracts (chloroform, acetone, ethanol and aqueous) of Nauclea orientalis leaves were evaluated separately on adult Indian earthworm (Pheretima posthuma) and compared with that of albendazole. It was found that the extracts exhibited, respectively dose-dependent action and inhibition of spontaneous motility (paralysis) and death of earthworms. The results indicated that the chloroform, ethyl acetate and ethanol extracts were more potent. PMID:21218070

  3. In vitro antimicrobial activity of Persian shallot (Allium hirtifolium).

    PubMed

    Soroush, Setareh; Taherikalani, Morovat; Asadollahi, Parisa; Asadollahi, Khairollah; Taran, Mojtaba; Emaneini, Mohammad; Alizadeh, Sajjad

    2012-01-01

    Allium hirtifolium is a Persian native plant grown in cool mountain slopes of Iran. It has been used as a spice in Iran for many years. According to the literature review, there are no considerable reports on the antimicrobial properties of this plant. In this study, the antimicrobial activity of Persian shallot hydroalcoholic extract and F1 fraction of the plant (containing amino acid derivatives and/or other cationic compounds) was investigated on some Gram positive cocci and bacilli, Gram negative bacilli, two protozoa, a yeast and a fungus. Excellent activity against Candida albicans (MIC = 64 microg/ml, MBC = 128 microg/ml), Leishmania infantum (MIC = 0.2 mg/ml on the first day of study) and Trichomonas vaginalis (MIC = 5 microg/ml in PSDE form) and a moderate activity against Bacillus spp and Pseudomonas aeroginosa (MIC = 128 microg/ml) was observed. The results showed that this plant contains some anti-trichomonas and anti-leishmania components. PMID:23210319

  4. Urokinase-type plasminogen activator does not affect in vitro bovine embryo development and quality.

    PubMed

    Krania, Fotini; Dovolou, Eleni; Rekkas, Constantinos A; Heras, Sonia; Pappas, Ioannis; Soom, Ann Van; Amiridis, Georgios S

    2015-06-01

    The effects of modification of the in vitro embryo culture media (IVC) with the addition of urokinase-type plasminogen activator (u-PA) on the yield and/or quality of bovine embryos were examined in two experiments. In Experiment 1, denuded embryos were cultured in semi-defined synthetic oviductal fluid (SOF) for seven days, while in Experiment 2 embryos were co-cultured with cumulus cell monolayer in a serum-containing SOF medium. Plasminogen activator activity (PAA) and plasminogen activator inhibition (PAI) were determined in all spent IVC media. At the activity used (5 IU/ml), u-PA had no effect either on in vitro embryo production rates or on embryo quality as revealed by gene expression analysis of 10 important mRNA transcripts related to apoptosis, oxidation, implantation and metabolism. PAA and PAI analysis indicated the need for wellbalanced plasminogen activators and inhibitors as a culture environment for embryo development. However, more research is needed to unveil the mechanism by which u-PA is involved in in vitro embryo production systems. PMID:26051263

  5. Nitroxoline impairs tumor progression in vitro and in vivo by regulating cathepsin B activity

    PubMed Central

    Mitrovi?, Ana; Sosi?, Izidor; Gobec, Stanislav; Vasiljeva, Olga; Turk, Boris; ?emažar, Maja; Serša, Gregor; Kos, Janko

    2015-01-01

    Cathepsin B is a ubiquitously expressed lysosomal cysteine protease that participates in protein turnover within lysosomes. However, its protein and activity levels have been shown to be increased in cancer. Cathepsin B endopeptidase activity is involved in the degradation of extracellular matrix, a process that promotes tumor invasion, metastasis and angiogenesis. Previously, we reported an established antibiotic nitroxoline as a potent and selective inhibitor of cathepsin B. In the present study, we elucidated its anti-tumor properties in in vitro and in vivo tumor models. Tumor and endothelial cell lines with high levels of active cathepsin B were selected for functional analysis of nitroxoline in vitro. Nitroxoline significantly reduced extracellular DQ-collagen IV degradation by all evaluated cancer cell lines using spectrofluorimetry. Nitroxoline also markedly decreased tumor cell invasion monitored in real time and reduced the invasive growth of multicellular tumor spheroids, used as a 3D in vitro model of tumor invasion. Additionally, endothelial tube formation was significantly reduced by nitroxoline in an in vitro angiogenesis assay. Finally, nitroxoline significantly abrogated tumor growth, angiogenesis and metastasis in vivo in LPB fibrosarcoma and MMTV-PyMT breast cancer mouse models. Overall, our results designate nitroxoline as a promising drug candidate for anti-cancer treatment. PMID:25848918

  6. Review on in vivo and in vitro methods evaluation of antioxidant activity

    PubMed Central

    Alam, Md. Nur; Bristi, Nusrat Jahan; Rafiquzzaman, Md.

    2012-01-01

    A good number of abstracts and research articles (in total 74) published, so far, for evaluating antioxidant activity of various samples of research interest were gone through where 407 methods were come across, which were repeated from 29 different methods. These were classified as in vitro and in vivo methods. And those are described and discussed below in this review article. In the later part of this review article, frequency of in vitro as well as in vivo methods is analyzed with a bar diagram. Solvents are important for extracting antioxidants from natural sources. Frequency of solvents used for extraction is also portrayed and the results are discussed in this article. As per this review there are 19 in vitro methods and 10 in vivo methods that are being used for the evaluation of antioxidant activity of the sample of interest. DPPH method was found to be used mostly for the in vitro antioxidant activity evaluation purpose while LPO was found as mostly used in vivo antioxidant assay. Ethanol was with the highest frequency as solvent for extraction purpose. PMID:24936134

  7. The lipid moiety of brincidofovir is required for in vitro antiviral activity against Ebola virus.

    PubMed

    McMullan, Laura K; Flint, Mike; Dyall, Julie; Albarińo, César; Olinger, Gene G; Foster, Scott; Sethna, Phiroze; Hensley, Lisa E; Nichol, Stuart T; Lanier, E Randall; Spiropoulou, Christina F

    2016-01-01

    Brincidofovir (BCV) is the 3-hexadecyloxy-1-propanol (HDP) lipid conjugate of the acyclic nucleoside phosphonate cidofovir (CDV). BCV has established broad-spectrum activity against double-stranded DNA (dsDNA) viruses; however, its activity against RNA viruses has been less thoroughly evaluated. Here, we report that BCV inhibited infection of Ebola virus in multiple human cell lines. Unlike the mechanism of action for BCV against cytomegalovirus and other dsDNA viruses, phosphorylation of CDV to the diphosphate form appeared unnecessary. Instead, antiviral activity required the lipid moiety and in vitro activity against EBOV was observed for several HDP-nucleotide conjugates. PMID:26526586

  8. Similar activity of mecamylamine stereoisomers in vitro and in vivo

    PubMed Central

    Papke, Roger L.; Stokes, Clare; Muldoon, Pretal; Damaj, M. Imad

    2014-01-01

    A previous characterization of mecamylamine stereoisomers using nicotinic acetylcholine receptors expressed in Xenopus oocytes revealed only small differences between the activity of the R and S forms of mecamylamine. However, that work was limited in the breadth of receptor subtypes tested, especially in regard to the discrimination of high and low sensitivity receptors, which differ in the ratios of alpha and beta subunits. We report new data using subunit concatamers, which produce uniform populations of high-sensitivity or low-sensitivity receptors, as well as alpha2, alpha5, and alpha6-containing receptors, which were not studied previously. Consistent with previous studies, we found that beta4-containing receptors were most sensitive to mecamylamine and that the IC50 values for the inhibition of net charge were lower than for inhibition of peak currents. No large differences were seen between the activities of the mecamylamine isomers. Additionally, a previously reported potentiation of high-sensitivity ?4?2 receptors by S-mecamylamine could not be reproduced in the oocyte system, even with mutants that had greatly reduced sensitivity to mecamylamine inhibition or when the selective agonist TC-2559 was used. In vivo studies suggested that the R-isomer might be somewhat more potent than the S isomer at blocking CNS effects of nicotine. Although the potency difference was no more than a factor of two, it is consistent with lower LD50 estimates previously reported for the R isomer. Our results significantly extend knowledge of the nicotinic acetylcholine receptor activity profile of mecamylamine and support the hypothesis that these effects are not strongly stereoisomer selective. PMID:24161916

  9. Aldosterone Activates Transcription Factor Nrf2 in Kidney Cells Both In Vitro and In Vivo

    PubMed Central

    Oteiza, Patricia I.; Link, Samuel; Hey, Valentin; Stopper, Helga; Schupp, Nicole

    2014-01-01

    Abstract Aims: An increased kidney cancer risk was found in hypertensive patients, who frequently exhibit hyperaldosteronism, known to contribute to kidney injury, with oxidative stress playing an important role. The capacity of kidney cells to up-regulate transcription factor nuclear factor-erythroid-2-related factor 2 (Nrf2), a key regulator of the cellular antioxidative defense, as a prevention of aldosterone-induced oxidative damage was investigated both in vitro and in vivo. Results: Aldosterone activated Nrf2 and increased the expression of enzymes involved in glutathione (GSH) synthesis and detoxification. This activation depended on the mineralocorticoid receptor (MR) and oxidative stress. In vitro, Nrf2 activation, GSH amounts, and target gene levels decreased after 24?h, while oxidant levels remained high. Nrf2 activation could not protect cells against oxidative DNA damage, as aldosterone-induced double-strand breaks and 7,8-dihydro-8-oxo-guanine (8-oxodG) lesions steadily rose. The Nrf2 activator sulforaphane enhanced the Nrf2 response both in vitro and in vivo, thereby preventing aldosterone-induced DNA damage. In vivo, Nrf2 activation further had beneficial effects on the aldosterone-caused blood pressure increase and loss of kidney function. Innovation: This is the first study showing the activation of Nrf2 by aldosterone. Moreover, the results identify sulforaphane as a substance that is capable of preventing aldosterone-induced damage both in vivo and in vitro. Conclusion: Aldosterone-induced Nrf2 adaptive response cannot neutralize oxidative actions of chronically increased aldosterone, which, therefore could be causally involved in the increased cancer incidence of hypertensive individuals. Enhancing the cellular antioxidative defense with sulforaphane might exhibit beneficial effects. Antioxid. Redox Signal. 21, 2126–2142. PMID:24512358

  10. In vitro and in vivo Bone-Forming Activity of Saururus chinensis Extract.

    PubMed

    Moon, Seong-Hee; Choi, Sik-Won; Park, Sang-Joon; Ryu, Shi-Yong; Hwang, Kyu-Seok; Kim, Cheol-Hee; Kim, Seong Hwan

    2015-07-01

    Bone is maintained by osteoclast-mediated resorption and osteoblast-mediated formation. Recently, anti-osteoporotic activity of Saururus chinensis extract (SCE) and anti-osteoclastogenic activity of its components have been reported, but the effect of SCE on bone formation has not been studied well. Therefore, in this study, we investigated whether Saururus chinensis SCE exhibits in vitro osteogenic and in vivo bone-forming activity. extract strongly enhanced the bone morphogenetic protein (BMP)-2-stimulated induction of alkaline phosphatase, an early phase biomarker of osteoblast differentiation, in bi-potential mesenchymal progenitor C2C12 cells. In vitro osteogenic activity of SCE was accompanied by enhanced expression of BMP-2, BMP-4, BMP-7 and BMP-9 mRNA. In addition, a pharmacological inhibition study suggested the involvement of p38 activation in the osteogenic action of SCE. Moreover, the BMP dependency and the involvement of p38 activation in the osteogenic action of SCE were confirmed by the treatment of noggin, an antagonist of BMP. Saururus chinensis extract also exhibited to induce runt-related transcription factor 2 activation at the high concentration. Furthermore, the in vivo osteogenic activity of SCE was confirmed in zebrafish and mouse calvarial bone formation models, suggesting the possibility of its use for bone formation. In conclusion, we suggested that in vivo anti-osteoporotic activity of SCE could be because of its dual action in bone, anti-osteoclastogenic and anabolic activity. PMID:25869918

  11. Antagonistic Activity of Lactobacillus Isolates against Salmonella typhi In Vitro

    PubMed Central

    Abdel-Daim, Amira; Hassouna, Nadia; Hafez, Mohamed; Ashor, Mohamed Seif Aldeen; Aboulwafa, Mohammad M.

    2013-01-01

    Background. Enteric fever is a global health problem, and rapidly developing resistance to various drugs makes the situation more alarming. The potential use of Lactobacillus to control typhoid fever represents a promising approach, as it may exert protective actions through various mechanisms. Methods. In this study, the probiotic potential and antagonistic activities of 32 Lactobacillus isolates against Salmonella typhi were evaluated. The antimicrobial activity of cell free supernatants of Lactobacillus isolates, interference of Lactobacillus isolates with the Salmonella adherence and invasion, cytoprotective effect of Lactobacillus isolates, and possibility of concurrent use of tested Lactobacillus isolates and antibiotics were evaluated by testing their susceptibilities to antimicrobial agents, and their oxygen tolerance was also examined. Results. The results revealed that twelve Lactobacillus isolates could protect against Salmonella typhi infection through interference with both its growth and its virulence properties, such as adherence, invasion, and cytotoxicity. These Lactobacillus isolates exhibited MIC values for ciprofloxacin higher than those of Salmonella typhi and oxygen tolerance and were identified as Lactobacillus plantarum. Conclusion. The tested Lactobacillus plantarum isolates can be introduced as potential novel candidates that have to be subjected for in vivo and application studies for treatment and control of typhoid fever. PMID:24191248

  12. Spontaneous Electrical Activity in the Human Fetal Cortex in vitro

    PubMed Central

    Moore, Anna R.; Zhou, Wen-Liang; Jakovcevski, Igor; Zecevic, Nada; Antic, Srdjan D.

    2011-01-01

    Our knowledge about the developing human cerebral cortex is based on the analysis of fixed postmortem material. Here we utilize electrical recordings from unfixed human postmortem tissue to characterize the synaptic physiology and spontaneous network activity of pioneer cortical neurons (“subplate neurons”). Our electrophysiological experiments show that functional glutamate or GABA ionotropic receptors are expressed on human subplate (SP) neurons as early as 20 gestational weeks. Extracellular (synaptic) stimulations evoked postsynaptic potentials in a very small fraction of SP neurons, suggesting that functional synaptic contacts are rare at mid-gestation. Although synaptic inputs were scarce, we regularly observed spontaneous (unprovoked) electrical activity among human SP neurons, comprised of sustained plateau depolarizations and bursts of action potential firing, which resembled cortical UP and DOWN states in the adult neocortex. Plateau depolarizations and bursts of AP firing are thought to depend on the mature morphology and physiology of adult cortical network. However, our current data reveal that similar cortical rhythm is generated by a very immature ensemble of human fetal neurons. In the relative absence of sensory inputs, as in development in utero, or in slow wave sleep (i.e. throughout the entire lifespan), the spontaneous slow oscillatory pattern (UP and DOWN states) is a fundamental aspect of human cortical physiology. PMID:21325506

  13. In vitro - in vivo correlations for endocrine activity of a mixture of currently used pesticides

    SciTech Connect

    Taxvig, Camilla; Hadrup, Niels; Boberg, Julie; Axelstad, Marta; Bossi, Rossana; Bonefeld-Jřrgensen, Eva Cecilie; Vinggaard, Anne Marie

    2013-11-01

    Two pesticide mixtures were investigated for potential endocrine activity. Mix 3 consisted of bitertanol, propiconazole, and cypermethrin, and Mix 5 included malathion and terbuthylazine in addition to the three pesticides in Mix 3. All five single pesticides and the two mixtures were investigated for their ability to affect steroidogenesis in vitro in H295R cells. The pesticides alone and both mixtures affected steroidogenesis with both mixtures causing increase in progesterone and decrease in testosterone. For Mix 5 an increase in estradiol was seen as well, indicating increased aromatase activity. The two mixtures were also investigated in pregnant rats dosed from gestational day 7 to 21, followed by examination of dams and fetuses. Decreased estradiol and reduced placental testosterone were seen in dams exposed to Mix 5. Also a significant increase in aromatase mRNA-levels in female adrenal glands was found for Mix5. However, either of the two mixtures showed any effects on fetal hormone levels in plasma or testis, or on anogenital distance. Overall, potential aromatase induction was found for Mix 5 both in vitro and in vivo, but not for Mix 3, an effect likely owed to terbuthylazine in Mix 5. However, the hormonal responses in vitro were only partly reflected in vivo, probably due to some toxicokinetic issues, as the pesticide levels in the amniotic fluid also were found to be negatively affected by the number of compounds present in the mixtures. Nonetheless, the H295R assay gives hints on conceivable interference with steroidogenesis, thus generating hypotheses on in vivo effects. - Highlights: • The study examines the endocrine disrupting potential of mixtures of pesticides. • All single pesticides and both mixtures affected steroidogenesis in vitro. • Potential aromatase induction was found for Mix 5 both in vitro and in vivo. • The hormonal responses in vitro were only partly reflected in vivo.

  14. In vitro activity of colistin sulfate against Enterobacteriaceae producing extended-spectrum ?-lactamases.

    PubMed

    Ku, Yee-Huang; Lee, Mei-Feng; Chuang, Yin-Ching; Chen, Chi-Chung; Yu, Wen-Liang

    2015-12-01

    The widespread multidrug-resistant Enterobacteriaceae pose a serious therapeutic challenge. Colistin and tigecycline are potential antimicrobial agents for treating infections caused by extended-spectrum ?-lactamase (ESBL)-producing Enterobacteriaceae. We evaluated the in-vitro activity of colistin sulfate against 253 ESBL producers isolated from patients admitted to a medical center in southern Taiwan (Escherichia coli, n = 82; Klebsiella pneumoniae, n = 102; Enterobacter cloacae, n = 34; and Serratia marcescens, n = 35). Colistin showed promising in-vitro activity against E. coli, K. pneumoniae, and E. cloacae, but not S. marcescens. One ESBL-producing K. pneumoniae strain with resistance to carbapenems (ertapenem, imipenem, and meropenem) was selected for time-killing studies. A combination of colistin and tigecycline showed synergism, but there was an inoculum effect. In conclusion, colistin was active against most ESBL-producing Enterobacteriaceae, and a combination of colistin with tigecycline was synergistic against some highly resistant strains, even those with carbapenem resistance. PMID:24388585

  15. A dual functional peptide carrying in vitro selected catalytic and binding activities.

    PubMed

    Zhu, Liping; Wang, Wei; Zhao, Haixu; Xu, Muye; Tada, Seiichi; Uzawa, Takanori; Liu, Mingzhe; Ito, Yoshihiro

    2015-10-14

    When minimal functional sequences are used, it is possible to integrate multiple functions on a single peptide chain, like a "single stroke drawing". Here a dual functional peptide was designed by combining in vitro selected catalytic and binding activities. For catalytic activity, we performed in vitro selection for a peptide aptamer binding to hemin by using ribosome display and isolated a peptide that had peroxidase activity in the presence of hemin. By combining the selected catalytic peptide with a peptide antigen, which can be recognized by an antibody, an enzyme-antibody conjugate-like peptide was obtained. This study demonstrates a successful strategy to create dual functionalized peptide chains for use in immunoassays. PMID:26272651

  16. Immunomodulatory activity in vitro and in vivo of verbascose from mung beans (Phaseolus aureus).

    PubMed

    Dai, Zhuqing; Su, Di; Zhang, Yun; Sun, Yi; Hu, Bing; Ye, Hong; Jabbar, Saqib; Zeng, Xiaoxiong

    2014-11-01

    In the present study, the immunostimulatory activity of verbascose from mung beans (Phaseolus aureus) was evaluated by using in vitro cell models and in vivo animal models. The results of in vitro experiments showed that verbascose could enhance the ability of devouring neutral red of peritoneal macrophages and promote the release of nitric oxide and immune reactive molecules such as interleukin (IL)-6, IL-1?, interferon (IFN)-?, and IFN-?. Treatment with verbascose at a dose of 200 ?g/mL exhibited the best effects. For assay in vivo, administration of verbascose at a medium dose of 90 mg/kg body weight could significantly increase the index of spleen, activity of lysozyme in spleen and serum, hemolysin level in serum, and swelling rate of earlap in the delayed type of hypersensitivity (DTH) of immunosuppressed mice. All of the results suggested that verbascose had potent immunostimulatory activity and could be explored as a potential natural immunomodulatory agent in functional foods. PMID:25317918

  17. In vitro and in vivo anti-plasmodial activity of essential oils, including hinokitiol.

    PubMed

    Fujisaki, Ryuichi; Kamei, Kiyoko; Yamamura, Mariko; Nishiya, Hajime; Inouye, Shigeharu; Takahashi, Miki; Abe, Shigeru

    2012-03-01

    Abstract. The anti-plasmodial activity of 47 essential oils and 10 of their constituents were screened for in vitro activity against Plasmodium falciparum. Five of these essential oils (sandalwood, caraway, monarda, nutmeg, and Thujopsis dolabrata var. hondai) and 2 constituents (thymoquinone and hinokitiol) were found to be active against P. falciparum in vitro, with 50% inhibitory concentration (IC50) values equal to or less than 1.0 microg/ml. Furthermore, in vivo analysis using a rodent model confirmed the anti-plasmodial potential of subcutaneously administered sandalwood oil, and percutaneously administered hinokitiol and caraway oil against rodent P. berghei. Notably, these oils showed no efficacy when administered orally, intraperitoneally or intravenously. Caraway oil and hinokitiol dissolved in carrier oil, applied to the skin of hairless mice caused high levels in the blood, with concentrations exceeding their IC50 values. PMID:23082579

  18. Gamma radiation effects on phenolics, antioxidants activity and in vitro digestion of pistachio ( Pistachia vera) hull

    NASA Astrophysics Data System (ADS)

    Behgar, M.; Ghasemi, S.; Naserian, A.; Borzoie, A.; Fatollahi, H.

    2011-09-01

    The effect of gamma radiation (10, 20, 30, 40, 50 and 60 kGy) on tannin, total phenolics, antioxidants activity and in vitro digestion of pistachio hulls has been investigated in this study. The possibility of using the radial diffusion method based on software measurement of the rings area has also been investigated in this study. The software based method in radial diffusion method showed a higher r2 (0.995) value when compared to the traditional method. Irradiation reduced the tannin content ( P<0.01) and activity of antioxidants ( P<0.05) of pistachio hull extracts but increased the total phenolic content ( P<0.05). There was no effect of gamma irradiation on the in vitro digestion of the pistachio hull. Irradiation decreased the digestion rate of the pistachio hull at the dose of 40 kGy when compared to the control. This study showed that gamma irradiation decreased tannin and antioxidants activity of pistachio hull.

  19. Two different interictal spike patterns anticipate ictal activity in vitro.

    PubMed

    Avoli, Massimo; Panuccio, Gabriella; Herrington, Rochelle; D'Antuono, Margherita; de Guzman, Philip; Lévesque, Maxime

    2013-04-01

    4-Aminopyridine (4AP, 50 ?M) induces interictal- and ictal-like discharges in brain slices including parahippocampal areas such as the entorhinal cortex (EC) but the relation between these two types of epileptiform activity remains undifined. Here, by employing field potential recordings in rat EC slices during 4AP application, we found that: (i) interictal events have a wide range of duration (0.4-3.3 s) and interval of occurrence (1.4-84 s); (ii) ictal discharges are either preceded by an isolated "slow" interictal discharge (ISID; duration=1.5 ± 0.1s, interval of occurrence=33.8 ± 1.8 s) or suddenly initiate from a pattern of frequent polispike interictal discharge (FPID; duration=0.8 ± 0.1 s; interval of occurrence=2.7 ± 0.2 s); and (iii) ISID-triggered ictal events have longer duration (116 ± 7.3s) and interval of occurrence (425.8 ± 42.3 s) than those initiating suddenly during FPID (58.3 ± 7.8 s and 202.1 ± 21.8 s, respectively). Glutamatergic receptor antagonists abolished ictal discharges in all experiments, markedly reduced FPIDs but did not influence ISIDs. We also discovered that high-frequency oscillations (HFOs, 80-500 Hz) occur more frequently during ISIDs as compared to FPIDs, and mainly coincide with the onset of ISID-triggered ictal discharges. These findings indicate that interictal events may define ictal onset features resembling those seen in vivo in low-voltage fast activity onset seizures. We propose a similar condition to occur in vivo in temporal lobe epileptic patients and animal models. PMID:23270790

  20. Differential in vitro activities of ionophore compounds against Plasmodium falciparum and mammalian cells.

    PubMed Central

    Gumila, C; Ancelin, M L; Jeminet, G; Delort, A M; Miquel, G; Vial, H J

    1996-01-01

    Twenty-two ionophore compounds were screened for their antimalarial activities. They consisted of true ionophores (mobile carriers) and channel-forming quasi-ionophores with different ionic specificities. Eleven of the compounds were found to be extremely efficient inhibitors of Plasmodium falciparum growth in vitro, with 50% inhibitory concentrations of less than 10 ng/ml. Gramicidin D was the most active compound tested, with 50% inhibitory concentration of 0.035 ng/ml. Compounds with identical ionic specificities generally had similar levels of antimalarial activity, and ionophores specific to monovalent cations were the most active. Compounds were further tested to determine their in vitro toxicities against mammalian lymphoblast and macrophage cell lines. Nine of the 22 compounds, i.e., alborixin, lonomycin, nigericin, narasin, monensin and its methylated derivative, lasalocid and its bromo derivative, and gramicidin D, most specific to monovalent cations, were at least 35-fold more active in vitro against P. falciparum than against the two other mammalian cell lines. The enhanced ability to penetrate the erythrocyte membrane after infection could be a factor that determines ionophore selectivity for infected erythrocytes. PMID:8851578

  1. G17-modified hammerhead ribozymes are active in vitro and in vivo.

    PubMed

    Kalweit, Anne; Hammann, Christian

    2013-12-01

    Natural hammerhead ribozymes (HHRz) feature tertiary interactions between hairpin loops or bulges in two of three helices that surround the catalytic core of conserved nucleotides. Their conservation was originally established on minimal versions lacking the tertiary interactions. While those sequence requirements in general also apply to natural versions, we show here differences for the HHRz cleavage site N17. A guanosine at this position strongly impairs cleavage activity in minimal versions, whereas we observe for the G17 variants of four tertiary stabilized HHRz significant cleavage and ligation activity in vitro. Kinetic analyses of these variants revealed a reduced rate and extent of cleavage, compared with wild-type sequences, while variants with distorted tertiary interactions cleaved at a reduced rate, but to the same extent. Contrary to this, G17 variants exhibit similar in vitro ligation activity as compared with the respective wild-type motif. To also address the catalytic performance of these motifs in vivo, we have inserted HHRz cassettes in the lacZ gene and tested this ?-galactosidase reporter in Dictyostelium discoideum. In colorimetric assays, we observe differences in the enzymatic activity of ?-galactosidase, which correlate well with the activity of the different HHRz variants in vitro and which can be unambiguously attributed to ribozyme cleavage by primer extension analysis. PMID:24145822

  2. In vitro activities of natural products against oral Candida isolates from denture wearers

    PubMed Central

    2011-01-01

    Background Candida-associated denture stomatitis is a frequent infectious disease. Treatment of this oral condition is difficult because failures and recurrences are common. The aim of this study was to test the in vitro antifungal activity of pure constituents of essentials oils. Methods Eight terpenic derivatives (carvacrol, farnesol, geraniol, linalool, menthol, menthone, terpinen-4-ol, and ?-terpineol), a phenylpropanoid (eugenol), a phenethyl alcohol (tyrosol) and fluconazole were evaluated against 38 Candida isolated from denture-wearers and 10 collection Candida strains by the CLSI M27-A3 broth microdilution method. Results Almost all the tested compounds showed antifungal activity with MIC ranges of 0.03-0.25% for eugenol and linalool, 0.03-0.12% for geraniol, 0.06-0.5% for menthol, ?-terpineol and terpinen-4-ol, 0.03-0.5% for carvacrol, and 0.06-4% for menthone. These compounds, with the exception of farnesol, menthone and tyrosol, showed important in vitro activities against the fluconazole-resistant and susceptible-dose dependent Candida isolates. Conclusions Carvacrol, eugenol, geraniol, linalool and terpinen-4-ol were very active in vitro against oral Candida isolates. Their fungistatic and fungicidal activities might convert them into promising alternatives for the topic treatment of oral candidiasis and denture stomatitis. PMID:22118215

  3. Synthesis and Biological Evaluation of 1,4-Naphthoquinones and Quinoline-5,8-diones as Antimalarial and Schistosomicidal Agents

    PubMed Central

    Lanfranchi, Don Antoine; Cesar-Rodo, Elena; Bertrand, Benoît; Huang, Hsin-Hung; Day, Latasha; Johann, Laure; Elhabiri, Mourad; Becker, Katja; Williams, David L.

    2012-01-01

    Improving the solubility of polysubstituted 1,4-naphthoquinone derivatives was achieved by introducing nitrogen in two different positions of the naphthoquinone core, at C-5 and at C-8 of menadione through a two-step, straightforward synthesis based on the regioselective hetero-Diels-Alder reaction. The antimalarial and the antischistosomal activities of these polysubstituted aza-1,4-naphthoquinone derivatives were evaluated and led to the selection of distinct compounds for antimalarial versus antischistosomal action. The AgII-assisted oxidative radical decarboxylation of the phenyl acetic acids using AgNO3 and ammonium peroxodisulfate was modified to generate the 3-picolinyl-menadione with improved pharmacokinetic parameters, high antimalarial effects and capacity to inhibit the formation of ?-hematin. PMID:22777178

  4. Screening of selected medicinal plants for in vitro antidermatophytic activity.

    PubMed

    Kalaivanan, C; Chandrasekaran, M; Venkatesalu, V

    2013-12-01

    Different solvent extracts of leaves of Achyranthes aspera, Aegle marmelos, Cleistanthus collinus, Curcuma aromatica and Strychnos nux-vomica were screened against dermatophytes viz., Trichophyton mentagrophytes, T. rubrum, Microsporum gypseum, M. canis and Epidermophyton floccossum var. nigricans. The mean zones of inhibition were between 7.1 and 26.5mm. Minimum inhibitory concentrations (MIC) and minimum fungicidal concentrations (MFC) were from 7.81 to 500 and from 15.62 to 1000?g/mL respectively. The highest mean zone of inhibition (26.5mm), the lowest MIC value (7.81?g/ml) and the lowest MFC (15.62?g/ml) were observed in ethyl acetate extract of A. aspera against T. rubrum. The standard antifungal drug ketoconazole (10?g/disc) was used as the positive control and mean zones of inhibition were from 23 to 29mm. Further separation of active principle from ethyl acetate extract of A. aspera is under progress. PMID:24135649

  5. Structural features of endocrine active chemicals--A comparison of in vivo and in vitro data.

    PubMed

    Lewin, Geertje; Escher, Sylvia E; van der Burg, Bart; Simetska, Nelly; Mangelsdorf, Inge

    2015-08-01

    Studies on reproductive toxicity need high numbers of test animals. Therefore, we investigated whether chemical structural features (SF) in combination with in vitro data on specific adverse outcome pathways (AOPs) may be used for predicting reproductive toxicity of untested chemicals. Using the OECD Toolbox and expert judgment, we identified 89 structure groups for 275 chemicals for which the results of prenatal developmental toxicity or multigeneration studies were present in the Fraunhofer database on Fertility and Developmental Toxicity in experimental animals (FeDTex) database. Likewise, we evaluated 220 chemicals which had been tested in reporter gene assays on endocrine ((anti)estrogenic and (anti)androgenic) properties in the CALUX(®) test battery. There was a large spread of effect levels for substances within the chemical structure groups for both, in vivo and in vitro results. The groups of highest concern (diphenyl derivatives, planar conjugated systems with fused rings, phenols and organophosphates) correlated quite well, however, between the in vivo and in vitro data on estrogenic activity. For the 56 chemicals represented in both databases, lowest effect doses in vivo correlated well with the estrogenic activity in vitro. These results suggest that a panel of assays covering relevant AOPs and data on metabolism and toxicokinetics may allow prediction of relative reproductive or development toxicity potency within the identified chemical structure groups. PMID:25461902

  6. Comparative studies on biological activity of generic and branded enoxaparin in vivo and vitro.

    PubMed

    Tan, Xiaoqing; Cui, Huifei

    2015-10-01

    As per US Food and Drug Administration (FDA) requirement, the study was designed to conduct the fourth and fifth criteria of Abbreviated New Drug Application to demonstrate equivalence of generic and branded Enoxaparin in vivo and vitro.Pharmacodynamic behavior of branded and generic Enoxaparin was compared in a parallel study in rats based upon measurement of anti-FXa and anti-FIIa profiles. Blood samples collected at baseline and at 0.5, 1, 2, 4, 6, 8, 12 and 24 h postsubcutaneous administration of six batches of Lovenox and nine batches of generic Enoxaparin were evaluated for anti-FXa and anti-FIIa using chromogenic substrate method. Anti-FXa, Anti-FIIa, activated partial thromboplastin time (APTT), and Heptest prolongation time were conducted in vitro as per the United States Pharmacopeia method. Pharmacodynamics parameters were obtained including peak effect (anti-FXamax, anti-FIIamax), area under the effect curve (AUEC0-T and AUEC0-?), Tmax, and T1/2.Pharmacokinetic differences were not observed using anti-FXa or anti-FIIa. No statistically significant differences were observed between branded and generic Enoxaparin either in vitro anti-FXa, anti-FIIa, APTT, or Heptest assay.It can be concluded that they are bioequivalent in anticoagulant activity tested in vivo and vitro. PMID:26270263

  7. Antiviral Activity of Glycyrrhizin against Hepatitis C Virus In Vitro

    PubMed Central

    Matsumoto, Yoshihiro; Matsuura, Tomokazu; Aoyagi, Haruyo; Matsuda, Mami; Hmwe, Su Su; Date, Tomoko; Watanabe, Noriyuki; Watashi, Koichi; Suzuki, Ryosuke; Ichinose, Shizuko; Wake, Kenjiro; Suzuki, Tetsuro; Miyamura, Tatsuo; Wakita, Takaji; Aizaki, Hideki

    2013-01-01

    Glycyrrhizin (GL) has been used in Japan to treat patients with chronic viral hepatitis, as an anti-inflammatory drug to reduce serum alanine aminotransferase levels. GL is also known to exhibit various biological activities, including anti-viral effects, but the anti-hepatitis C virus (HCV) effect of GL remains to be clarified. In this study, we demonstrated that GL treatment of HCV-infected Huh7 cells caused a reduction of infectious HCV production using cell culture-produced HCV (HCVcc). To determine the target step in the HCV lifecycle of GL, we used HCV pseudoparticles (HCVpp), replicon, and HCVcc systems. Significant suppressions of viral entry and replication steps were not observed. Interestingly, extracellular infectivity was decreased, and intracellular infectivity was increased. By immunofluorescence and electron microscopic analysis of GL treated cells, HCV core antigens and electron-dense particles had accumulated on endoplasmic reticulum attached to lipid droplet (LD), respectively, which is thought to act as platforms for HCV assembly. Furthermore, the amount of HCV core antigen in LD fraction increased. Taken together, these results suggest that GL inhibits release of infectious HCV particles. GL is known to have an inhibitory effect on phospholipase A2 (PLA2). We found that group 1B PLA2 (PLA2G1B) inhibitor also decreased HCV release, suggesting that suppression of virus release by GL treatment may be due to its inhibitory effect on PLA2G1B. Finally, we demonstrated that combination treatment with GL augmented IFN-induced reduction of virus in the HCVcc system. GL is identified as a novel anti-HCV agent that targets infectious virus particle release. PMID:23874843

  8. E.coli Fis protein activates ribosomal RNA transcription in vitro and in vivo.

    PubMed Central

    Ross, W; Thompson, J F; Newlands, J T; Gourse, R L

    1990-01-01

    An upstream activation region (UAR) contributes to the extremely high activity of the Escherichia coli ribosomal RNA promoter, rrnB P1, increasing its activity 20- to 30-fold over that of the same promoter lacking the UAR. We have used DNase footprinting to define three specific sites in the rrnB P1 UAR that bind Fis, a protein identified previously by its role in recombinational enhancer function in other systems. We find that purified Fis activates transcription from promoters containing these sites 10- to 20-fold in vitro at concentrations correlating with the filling of these sites. Three approaches indicate that Fis contributes to the function of the UAR in vivo. First, there is a progressive loss in the activity of rrnB P1-lacZ fusions as Fis binding sites are deleted. Second, an rrnB P1 promoter with a mutation in a Fis binding site has 5-fold reduced transcription activity in vivo, dramatically reduced Fis binding in vitro, and shows no Fis dependent transcription activation in vitro. Third, upstream activation is reduced 5-fold in a Fis- strain. We show that rRNA promoters derepress in response to the loss of Fis in vivo in accord with the predictions of the negative feedback model for rRNA regulation. We find that fis is not essential for the function of two control systems known to regulate rRNA, growth rate dependent control and stringent control. On the basis of these results, we propose roles for Fis and the upstream activation system in rRNA synthesis. Images Fig.1 Fig.4 Fig.5 Fig.6 PMID:2209559

  9. Oxidative stress biomarkers and acetylcholinesterase activity in human erythrocytes exposed to clomazone (in vitro)

    PubMed Central

    Santi, Adriana; Menezes, Charlene; Duarte, Marta Maria F.; Leitemperger, Jossiele; Lópes, Thais; Loro, Vania L.

    2011-01-01

    The aim of this study was to investigate the effect of clomazone herbicide on oxidative stress biomarkers and acetylcholinesterase activity in human erythrocytes in in vitro conditions. The activity of catalase (CAT), superoxide dismutase (SOD) and acetylcholinesterase (AChE), as well as the levels of thiobarbituric acid reactive substances (TBARS) and reduced glutathione (GSH) were measured in human erythrocytes exposed (in vitro) to clomazone at varying concentrations in the range of 0, 100, 250 and 500 µg/L for 1 h at 37 °C.TBARS levels were significantly higher in erythrocytes incubated with clomazone at 100, 250 and 500 µg/L. However, erythrocyte CAT and AChE activities were decreased at all concentrations tested. SOD activity was increased only at 100 µg/L of clomazone. GSH levels did not change with clomazone exposure. These results clearly showed clomazone to induce oxidative stress and AChE inhibition in human erythrocytes (in vitro). We, thus, suggest a possible role of ROS on toxicity mechanism induced by clomazone in humans. PMID:22058656

  10. Cysticidal activity of extracts and isolated compounds from Teloxys graveolens: In vitro and in vivo studies.

    PubMed

    Palomares-Alonso, Francisca; Rojas-Tomé, Irma Susana; Juárez Rocha, Victorino; Palencia Hernández, Guadalupe; González-Maciel, Angélica; Ramos-Morales, Andrea; Santiago-Reyes, Rosalba; González-Hernández, Iliana Elvira; Jung-Cook, Helgi

    2015-09-01

    In the search of new alternatives for neurocysticercosis treatment, the cysticidal activity of organic extracts of Teloxys graveolens was evaluated. The in vitro activity of hexane, ethyl acetate and methanol extracts against Taenia crassiceps cysts was tested and the selectivity index relative to human fibroblasts was determined. Subsequently, the in vivo efficacy of the methanolic extract at doses of 200 and 500?mg/kg in the murine cysticercosis model was evaluated. The ultrastructural effects in vitro and in vivo of the methanolic extract were also investigated using scanning electron microscopy. Additionally, a bioassay-guided fractionation for the isolation of the cysticidal components was performed. Our in vitro findings revealed that all extracts exhibited good cysticidal activity with EC50 values from 44.8 to 67.1?µg/mL. Although the ethyl acetate and methanolic extracts displayed low cytotoxicity, the methanolic extract was the most selective. The methanolic extract also showed in vivo efficacy which was similar to that obtained with ABZ. Significant alterations were found on the germinal layer of the cysts, with a high accumulation of granules of glycogen and vacuoles. The bioguided fractionation of methanolic extract led to the isolation of three flavonoids: chrysin, pinocembrin and pinostrobin; among them, pinocembrin was the compound that displayed cysticidal activity. This is the first study which reveals that T. graveolens could be a potential source for cysticidal and non-toxic compounds. PMID:26072200

  11. Impact of human serum albumin on oritavancin in vitro activity against enterococci.

    PubMed

    McKay, Geoffrey A; Beaulieu, Sylvain; Sarmiento, Ingrid; Arhin, Francis F; Parr, Thomas R; Moeck, Gregory

    2009-06-01

    Oritavancin is a lipoglycopeptide with activity against gram-positive pathogens including vancomycin-resistant enterococci. The impact of human serum albumin (HSA) on oritavancin activity against enterococci was compared to those of vancomycin, daptomycin, teicoplanin, and linezolid in vitro using MIC and time-kill methods. Oritavancin MICs increased between 0- and 8-fold in the presence of HSA. In time-kill assays with HSA, oritavancin retained activity, killing or inhibiting enterococci more rapidly than did comparators when peak concentrations were simulated. PMID:19349514

  12. Impact of Human Serum Albumin on Oritavancin In Vitro Activity against Enterococci?

    PubMed Central

    McKay, Geoffrey A.; Beaulieu, Sylvain; Sarmiento, Ingrid; Arhin, Francis F.; Parr, Thomas R.; Moeck, Gregory

    2009-01-01

    Oritavancin is a lipoglycopeptide with activity against gram-positive pathogens including vancomycin-resistant enterococci. The impact of human serum albumin (HSA) on oritavancin activity against enterococci was compared to those of vancomycin, daptomycin, teicoplanin, and linezolid in vitro using MIC and time-kill methods. Oritavancin MICs increased between 0- and 8-fold in the presence of HSA. In time-kill assays with HSA, oritavancin retained activity, killing or inhibiting enterococci more rapidly than did comparators when peak concentrations were simulated. PMID:19349514

  13. In vitro activity of teichomycin and vancomycin alone and in combination with rifampin.

    PubMed

    Varaldo, P E; Debbia, E; Schito, G C

    1983-03-01

    The antibacterial activity of teichomycin, a glycopeptide antibiotic similar to vancomycin, has been evaluated in vitro and compared with that of vancomycin. Test strains included 130 staphylococci and 132 streptococci, with representatives of the major currently recognized species or groups, and lesser numbers of clostridia, propionibacteria, and group JK bacteria. Teichomycin was found to be more active than vancomycin. Its minimum inhibitory concentration (MIC) was two- to fourfold lower than that of vancomycin with staphylococci and anaerobic bacteria, and two- to eightfold lower with streptococci. No significant differences were observed with group JK bacteria. For most strains tested, minimum bactericidal concentrations (MBCs) of both teichomycin and vancomycin either equalled or exceeded by twofold the respective MICs. Higher MBC-to-MIC ratios were obtained for enterococci and pneumococci with both antibiotics. Both teichomycin and vancomycin showed similar in vitro interactions with rifampin in combination tests. Neither antagonism nor (with very few exceptions) synergism occurred. PMID:6221692

  14. Curcuminoid derivatives enhance telomerase activity in an in vitro TRAP assay.

    PubMed

    Taka, Thanachai; Changtam, Chatchawan; Thaichana, Pak; Kaewtunjai, Navakoon; Suksamrarn, Apichart; Lee, T Randall; Tuntiwechapikul, Wirote

    2014-11-15

    The length of telomeres controls the life span of eukaryotic cells. Telomerase maintains the length of telomeres in certain eukaryotic cells, such as germline cells and stem cells, and allows these cells to evade replicative senescence. Here, we report for the first time a number of curcuminoid derivatives that enhance telomerase activity in an in vitro TRAP assay. A preliminary analysis of structure-activity relationships found that the minimal requirement for this enhanced telomerase activity is a curcuminoid core with at least one n-pentylpyridine side chain, while curcuminoids with two such side chains exhibit even greater activity. The finding here might lead to a new class of telomerase activators that act directly or indirectly on telomerase, rather than through the reactivation of the telomerase reverse transcriptase (TERT) gene associated with other telomerase activators found in the literature. PMID:25305686

  15. Interlaboratory comparison of in vitro bioassays for screening of endocrine active chemicals in recycled water.

    PubMed

    Mehinto, Alvine C; Jia, Ai; Snyder, Shane A; Jayasinghe, B Sumith; Denslow, Nancy D; Crago, Jordan; Schlenk, Daniel; Menzie, Christopher; Westerheide, Sandy D; Leusch, Frederic D L; Maruya, Keith A

    2015-10-15

    In vitro bioassays have shown promise as water quality monitoring tools. In this study, four commercially available in vitro bioassays (GeneBLAzer(®) androgen receptor (AR), estrogen receptor-alpha (ER), glucocorticoid receptor (GR) and progesterone receptor (PR) assays) were adapted to screen for endocrine active chemicals in samples from two recycled water plants. The standardized protocols were used in an interlaboratory comparison exercise to evaluate the reproducibility of in vitro bioassay results. Key performance criteria were successfully achieved, including low background response, standardized calibration parameters and high intra-laboratory precision. Only two datasets were excluded due to poor calibration performance. Good interlaboratory reproducibility was observed for GR bioassay, with 16-26% variability among the laboratories. ER and PR bioactivity was measured near the bioassay limit of detection and showed more variability (21-54%), although interlaboratory agreement remained comparable to that of conventional analytical methods. AR bioassay showed no activity for any of the samples analyzed. Our results indicate that ER, GR and PR, were capable of screening for different water quality, i.e., the highest bioactivity was observed in the plant influent, which also contained the highest concentrations of endocrine active chemicals measured by LC-MS/MS. After advanced treatment (e.g., reverse osmosis), bioactivity and target chemical concentrations were both below limits of detection. Comparison of bioassay and chemical equivalent concentrations revealed that targeted chemicals accounted for ?5% of bioassay activity, suggesting that detection limits by LC-MS/MS for some chemicals were insufficient and/or other bioactive compounds were present in these samples. Our study demonstrated that in vitro bioassays responses were reproducible, and can provide information to complement conventional analytical methods for a more comprehensive water quality assessment. PMID:26177482

  16. In vitro propagation of the medicinal plant Ziziphora tenuior L. and evaluation of its antioxidant activity

    PubMed Central

    Dakah, Abdulkarim; Zaid, Salim; Suleiman, Mohamad; Abbas, Sami; Wink, Michael

    2014-01-01

    Ziziphora tenuior L. (Lamiaceae) is an aromatic herb used for its medicinal values against fungi, bacteria. Micropropagation can be used for large-scale multiplication of essential oil producing plants thus avoiding an overexploitation of natural resources. This work aims to develop a reliable protocol for the in vitro propagation of Z. tenuior, and to compare the antioxidant activity between in vitro propagated and wild plants. The explants were sterilized and cultured on MS medium containing different concentrations of growth regulators naphthalene acetic acid (NAA) or indole-3-butyric acid (IBA) with 0.5 mg/L of kinetin (Kin) callus formation was 70.2% after 45 days of incubation in dark on medium supplemented with 1.5 mg/L of NAA. After one month of callus culture on medium supplemented with 2 mg/L BA the shoot number was 5.12 and for the multiplication stage. The shoot number was 4.21 and length was 6.17 cm on medium supplemented with 1 mg/L Kin + 0.1 mg/L NAA. DPPH• reagent was used to test the antioxidant activity. The aqueous and methanol extracts of in vitro plants which were treated with 1.5 and 1 mg/L of kin plus 0.1 mg/L of NAA showed a strong DPPH• scavenging activity where IC50 was 0.307 and 0.369 mg/ml, respectively, while the IC50 of aqueous and methanol extracts of wild plants was 0.516 and 9.229 mg/ml, respectively. Our results suggested that plant growth regulators and in vitro culture conditions increased the antioxidant activity. PMID:25183942

  17. Activity of Imipenem against Klebsiella pneumoniae Biofilms In Vitro and In Vivo

    PubMed Central

    Chen, Ping; Seth, Akhil K.; Abercrombie, Johnathan J.; Mustoe, Thomas A.

    2014-01-01

    Encapsulated Klebsiella pneumoniae has emerged as one of the most clinically relevant and more frequently encountered opportunistic pathogens in combat wounds as the result of nosocomial infection. In this report, we show that imipenem displayed potent activity against established K. pneumoniae biofilms under both static and flow conditions in vitro. Using a rabbit ear model, we also demonstrated that imipenem was highly effective against preformed K. pneumoniae biofilms in wounds. PMID:24247132

  18. Novel Amino-pyrazole Ureas with Potent In Vitro and In Vivo Antileishmanial Activity.

    PubMed

    Mowbray, Charles E; Braillard, Stéphanie; Speed, William; Glossop, Paul A; Whitlock, Gavin A; Gibson, Karl R; Mills, James E J; Brown, Alan D; Gardner, J Mark F; Cao, Yafeng; Hua, Wen; Morgans, Garreth L; Feijens, Pim-Bart; Matheeussen, An; Maes, Louis J

    2015-12-24

    Visceral leishmaniasis is a severe parasitic disease that is one of the most neglected tropical diseases. Treatment options are limited, and there is an urgent need for new therapeutic agents. Following an HTS campaign and hit optimization, a novel series of amino-pyrazole ureas has been identified with potent in vitro antileishmanial activity. Furthermore, compound 26 shows high levels of in vivo efficacy (>90%) against Leishmania infantum, thus demonstrating proof of concept for this series. PMID:26571076

  19. In vitro propagation of the medicinal plant Ziziphora tenuior L. and evaluation of its antioxidant activity.

    PubMed

    Dakah, Abdulkarim; Zaid, Salim; Suleiman, Mohamad; Abbas, Sami; Wink, Michael

    2014-09-01

    Ziziphora tenuior L. (Lamiaceae) is an aromatic herb used for its medicinal values against fungi, bacteria. Micropropagation can be used for large-scale multiplication of essential oil producing plants thus avoiding an overexploitation of natural resources. This work aims to develop a reliable protocol for the in vitro propagation of Z. tenuior, and to compare the antioxidant activity between in vitro propagated and wild plants. The explants were sterilized and cultured on MS medium containing different concentrations of growth regulators naphthalene acetic acid (NAA) or indole-3-butyric acid (IBA) with 0.5 mg/L of kinetin (Kin) callus formation was 70.2% after 45 days of incubation in dark on medium supplemented with 1.5 mg/L of NAA. After one month of callus culture on medium supplemented with 2 mg/L BA the shoot number was 5.12 and for the multiplication stage. The shoot number was 4.21 and length was 6.17 cm on medium supplemented with 1 mg/L Kin + 0.1 mg/L NAA. DPPH• reagent was used to test the antioxidant activity. The aqueous and methanol extracts of in vitro plants which were treated with 1.5 and 1 mg/L of kin plus 0.1 mg/L of NAA showed a strong DPPH• scavenging activity where IC50 was 0.307 and 0.369 mg/ml, respectively, while the IC50 of aqueous and methanol extracts of wild plants was 0.516 and 9.229 mg/ml, respectively. Our results suggested that plant growth regulators and in vitro culture conditions increased the antioxidant activity. PMID:25183942

  20. Delayed cerebrospinal fluid sterilization, in vitro bactericidal activities, and side effects of selected beta-lactams.

    PubMed

    Dajani, A S; Pokowski, L H

    1990-01-01

    Ampicillin (or penicillin G) plus chloramphenicol, cefuroxime, ceftriaxone, and cefotaxime have been used in the treatment of bacterial meningitis beyond the neonatal period. Review of recent data from the USA and Europe indicates that delayed CSF sterilization occurs significantly more often with ampicillin/chloramphenicol and cefuroxime than with ceftriaxone and cefotaxime. Delayed CSF sterilization is associated with an increased morbidity and neurological complications. Previously reported in vitro interactions between chloramphenicol and various beta-lactam antibiotics indicate that for bacteria where chloramphenicol is only bacteriostatic, the combination of chloramphenicol with beta-lactams is antagonistic. Killing rates of various beta-lactams were compared against a number of gram-positive and gram-negative bacteria. Cidal activity of some beta-lactams was inoculum dependent. There was a good correlation between in vitro activity and ability to sterilize CSF. Ceftriaxone is highly protein bound and its use in newborns is discouraged. Diarrhea occurs significantly more often after cefriaxone use than after the use of other agents. Ceftriaxone is uniquely associated with a high frequency of biliary pseudolithiasis which may be symptomatic and can cause measureable morbidity. In selecting the "proper" antimicrobial agent for the treatment of bacterial meningitis considerations should be given to proven clinical efficacy, prompt CSF sterilization, rapid in vitro cidal activity, safety and cost. We recommend cefotaxime as the agent of choice in the treatment of bacterial meningitis. PMID:2091255

  1. Anti-glycated and antiradical activities in vitro of polysaccharides from Ganoderma capense

    PubMed Central

    Yan, Chunyan; Kong, Fansheng; Zhang, Dezhi; Cui, Jiangxia

    2013-01-01

    Background Ganoderma capense is a Ganoderma species and is widely used, especially in Asia, as a well-known medicinal mushroom for health-promoting effect and for treatment of chronic diseases, such as diabetes, aging, etc. G. capense is rich of polysaccharide. Objective: To isolate the polysaccharides from G. capense and evaluate their anti-glycated and antiradical activities in vitro. Materials and Methods The dried powder of submerged fermentation culturing mycelium of G. capense was defatted, extracted with water/alkaline water followed by ethanol precipitation and deproteinated. And four crude polysaccharides, named as GC50, GC70, GC90 and GCB, were obtained. For the first time, the in vitro anti-glycated activities of the four samples were studied by non-enzymatic glycation reaction. Then, the DPPH radical and hydroxyl radical assays were established to estimate the antiradical capacity of the four samples. Meanwhile the contents of polysaccharides were determined by phenol-sulphuric acid colorimetry. Results and Conclusion Preliminary antiradical in vitro studies indicated that the four crude polysaccharides showed concentration-dependent scavenging abilities on DPPH and hydroxyl radicals. The evaluation of anti-glycation activity suggested that GC70 had good potential for inhibiting the formation of advanced glycation end products. Time- and dose-dependent effects were also observed for all GC70 samples. PMID:23661989

  2. Quantitative studies on the in vitro metabolic activation of dimethylnitrosamine by rat liver postmitochondrial supernatant

    SciTech Connect

    Doolittle, D.J.; Goodman, J.I.

    1984-08-01

    The metabolic activation of dimethylnitrosamine (DMN) to mutagenic and/or cytotoxic intermediates in vitro has been characterized and the relationship between DMN demethylase and ethoxyresorufin-O-deethylase (EROD) or ethylmorphine-N-demethylase (EMND) has been evaluated. A mammalian assay system which uses the postmitochondrial supernatant (S-15 fraction) prepared from a rat liver homogenate as an enzyme source and V79 Chinese hamster cells as targets for chemically induced damage was used. The enzyme pattern of the S-15 fraction was altered by pretreatment of experimental animals in vivo and/or by the use of enzyme inhibitors in vitro. The results of these studies indicate that the concentration of S-15 fraction in the reaction mixture can markedly influence the degree of DMN-induced cytotoxicity when it is metabolized in vitro and that the degree of DMN-induced cytotoxicity and mutagenicity are linearly related. The degree of cytotoxicity and mutagenicity induced in V79 cells by DMN does not correlate with EROD activity (a measure of 3-methylcholanthrene-inducible mixed-function oxidases) nor with EMND activity (a measure of phenobarbital-inducible mixed function oxidases) in the S-15 fraction. 28 references, 4 figures.

  3. Quantitative studies on the in vitro metabolic activation of dimethylnitrosamine by rat liver postmitochondrial supernatant.

    PubMed Central

    Doolittle, D J; Goodman, J I

    1984-01-01

    The metabolic activation of dimethylnitrosamine (DMN) to mutagenic and/or cytotoxic intermediates in vitro has been characterized and the relationship between DMN demethylase and ethoxyresorufin-O-deethylase (EROD) or ethylmorphine-N-demethylase (EMND) has been evaluated. A mammalian assay system which uses the postmitochondrial supernatant (S-15 fraction) prepared from a rat liver homogenate as an enzyme source and V79 Chinese hamster cells as targets for chemically induced damage was used. The enzyme pattern of the S-15 fraction was altered by pretreatment of experimental animals in vivo and/or by the use of enzyme inhibitors in vitro. The results of these studies indicate that the concentration of S-15 fraction in the reaction mixture can markedly influence the degree of DMN-induced cytotoxicity when it is metabolized in vitro and that the degree of DMN-induced cytotoxicity and mutagenicity are linearly related. The degree of cytotoxicity and mutagenicity induced in V79 cells by DMN does not correlate with EROD activity (a measure of 3-methylcholanthrene-inducible mixed-function oxidases) nor with EMND activity (a measure of phenobarbital-inducible mixed function oxidases) in the S-15 fraction. PMID:6499815

  4. Potent Inhibitors of Plasmodium Phospholipid Metabolism with a Broad Spectrum of In Vitro Antimalarial Activities

    PubMed Central

    Ancelin, Marie L.; Calas, Michčle; Vidal-Sailhan, Valérie; Herbuté, Serge; Ringwald, Pascal; Vial, Henri J.

    2003-01-01

    We characterized the potent in vitro antimalarial activity and biologic assessment of 13 phospholipid polar head analogs on a comparative basis. There was a positive relationship between the abilities of the drugs to inhibit parasite growth in culture and their abilities to specifically inhibit phosphatidylcholine biosynthesis of Plasmodium falciparum-infected erythrocytes. Maximal activity of G25 was observed for the trophozoite stage of the 48-h erythrocytic cycle (50% inhibitory concentration, 0.75 nM), whereas the schizont and ring stages were 12- and 213-fold less susceptible. The compounds exerted a rapid nonreversible cytotoxic effect, with complete clearance of parasitemia after 5 h of contact with the mature stages. The compounds were highly specific against P. falciparum, with much lower toxicity against three other mammalian cell lines, and the in vitro therapeutic indices ranged from 300 to 2,500,000. Finally, the monoquaternary ammonium E10 and two bis-ammonium salts, G5 and G25, were similarly active against multiresistant strains and fresh isolates of P. falciparum. This impressive selective in vitro toxicity against P. falciparum strongly highlights the clinical potential of these quaternary ammonium salts for malarial chemotherapy. PMID:12878524

  5. Effect of centrifugation on early embryonic development and parthenogenetic activation of bovine oocytes matured in vitro.

    PubMed

    Chung, J T; Downey, B R; Casper, R F; Chian, R C

    2001-01-01

    This study examined the fertilization, early developmental competence and capacity for parthenogenetic activation of bovine oocytes matured in vitro after centrifugation. Immature oocytes were cultured in tissue culture medium 199 supplemented with 10% fetal bovine serum and 75 mIU mL(-1) FSH + LH at 5% CO2 to facilitate maturation. After culture for 24 or 30 h, the metaphase-II stage oocytes were centrifuged at 3000, 5000, 7000 or 10000g for 5 min before in vitro fertilization or parthenogenetic activation. Frozen-thawed bull semen was used for in vitro fertilization. For parthenogenetic activation, the oocytes were exposed to 20 microM calcium ionophore A23187 for 5 min at room temperature. Fertilization rates were not different between control and treatment groups (87.7% v. 74.6%, 73.4%, 75.9% and 76.4% respectively). Also, there were no differences in early embryonic development between control and treatment groups (rates of blastocyst formation were 21.1% v 20.2%, 28.8%, 31.2% and 24.1% respectively). When the oocytes were centrifuged at various speeds alone, the activation rate of oocytes was significantly higher (P < 0.05) in the 10000g treatment group compared with control (10.8% v 0.0%). There were no differences in the activation rates of oocytes between control and treatment groups at speeds up to 7000g (70.9% v. 71.9%, 78.3% and 77.2% respectively) after centrifugation and stimulation with Ca(2+)-ionophore. However, the activation rate of oocytes was significantly higher (P < 0.05) in the 10000g treatment group compared with control (70.9% v. 83.1%). In addition, the percentage of activated oocytes with diploid formation was significantly higher in the oocytes after centrifugation at 10000g and stimulation with calcium ionophore A23187 than in the control (18.4% v 7.1%). These results indicate that centrifugation of oocytes matured in vitro has no detrimental effect on fertilization and subsequent early embryonic development. They also indicate that the oocytes might be parthenogenetically activated after centrifugation and that high-speed centrifugation may induce activation of some oocytes. The results suggest that the optimal speed for centrifugation of bovine oocytes might be < or = 7000g to enhance the visibility of nuclear elements for further micromanipulation. PMID:11833934

  6. Hilar somatostatin interneurons contribute to synchronized GABA activity in an in vitro epilepsy model.

    PubMed

    Grosser, Sabine; Queenan, Bridget N; Lalchandani, Rupa R; Vicini, Stefano

    2014-01-01

    Epilepsy is a disorder characterized by excessive synchronized neural activity. The hippocampus and surrounding temporal lobe structures appear particularly sensitive to epileptiform activity. Somatostatin (SST)-positive interneurons within the hilar region have been suggested to gate hippocampal activity, and therefore may play a crucial role in the dysregulation of hippocampal activity. In this study, we examined SST interneuron activity in the in vitro 4-aminopyridine (4-AP) model of epilepsy. We employed a multi-disciplinary approach, combining extracellular multi-electrode array (MEA) recordings with patch-clamp recordings and optical imaging using a genetically encoded calcium sensor. We observed that hilar SST interneurons are strongly synchronized during 4-AP-induced local field potentials (LFPs), as assayed by Ca(2+) imaging as well as juxtacellular or intracellular recording. SST interneurons were particularly responsive to GABA-mediated LFPs that occurred in the absence of ionotropic glutamatergic transmission. Our results present evidence that the extensive synchronized activity of SST-expressing interneurons contribute to the generation of GABAergic LFPs in an in vitro model of temporal lobe seizures. PMID:24465989

  7. Linarin Inhibits the Acetylcholinesterase Activity In-vitro and Ex-vivo

    PubMed Central

    Feng, Xinchi; Wang, Xin; Liu, Youping; Di, Xin

    2015-01-01

    Linarin is a flavone glycoside in the plants Flos chrysanthemi indici, Buddleja officinalis, Cirsium setosum, Mentha arvensis and Buddleja davidii, and has been reported to possess analgesic, antipyretic, anti-inflammatory and neuroprotective activities. In this paper, linarin was investigated for its AChE inhibitory potential both in-vitro and ex-vivo. Ellman’s colorimetric method was used for the determination of AChE inhibitory activity in mouse brain. In-vitro assays revealed that linarin inhibited AChE activity with an IC50 of 3.801 ± 1.149 ?M. Ex-vivo study showed that the AChE activity was significantly reduced in both the cortex and hippocampus of mice treated intraperitoneally with various doses of linarin (35, 70 and 140 mg/Kg). The inhibition effects produced by high dose of linarin were the same as that obtained after huperzine A treatment (0.5 mg/Kg). Molecular docking study revealed that both 4’-methoxyl group and 7-O-sugar moiety of linarin played important roles in ligand-receptor binding and thus they are mainly responsible for AChE inhibitory activity. In view of its potent AChE inhibitory activity, linarin may be a promising therapeutic agent for the treatment of some diseases associated with AChE, such as glaucoma, myasthenia gravis, gastric motility and Alzheimer’s disease. PMID:26330885

  8. Linarin Inhibits the Acetylcholinesterase Activity In-vitro and Ex-vivo.

    PubMed

    Feng, Xinchi; Wang, Xin; Liu, Youping; Di, Xin

    2015-01-01

    Linarin is a flavone glycoside in the plants Flos chrysanthemi indici, Buddleja officinalis, Cirsium setosum, Mentha arvensis and Buddleja davidii, and has been reported to possess analgesic, antipyretic, anti-inflammatory and neuroprotective activities. In this paper, linarin was investigated for its AChE inhibitory potential both in-vitro and ex-vivo. Ellman's colorimetric method was used for the determination of AChE inhibitory activity in mouse brain. In-vitro assays revealed that linarin inhibited AChE activity with an IC50 of 3.801 ± 1.149 ?M. Ex-vivo study showed that the AChE activity was significantly reduced in both the cortex and hippocampus of mice treated intraperitoneally with various doses of linarin (35, 70 and 140 mg/Kg). The inhibition effects produced by high dose of linarin were the same as that obtained after huperzine A treatment (0.5 mg/Kg). Molecular docking study revealed that both 4'-methoxyl group and 7-O-sugar moiety of linarin played important roles in ligand-receptor binding and thus they are mainly responsible for AChE inhibitory activity. In view of its potent AChE inhibitory activity, linarin may be a promising therapeutic agent for the treatment of some diseases associated with AChE, such as glaucoma, myasthenia gravis, gastric motility and Alzheimer's disease. PMID:26330885

  9. In vitro killing of Entamoeba histolytica trophozoites by interferon-gamma-activated mouse macrophages.

    PubMed

    Ghadirian, E; Bout, D T

    1988-03-01

    The effect of murine interferon gamma (IFN-gamma) on macrophage activation for amoebicidal activity was examined. Peritoneal macrophages were harvested from C57BL/6 mice and preincubated with IFN-gamma and/or lipopolysaccharide (LPS). In vitro amoebicidal activity of these macrophages was determined by trypan blue exclusion test against a virulent strain of E. histolytica (IP:0682:1). It was found that in vitro amoebicidal activity was evident in macrophage monolayers treated with both IFN-gamma and LPS. Macrophages treated with IFN-gamma alone did not develop cytotoxic activity unless they were exposed to LPS as a second triggering signal. The ability of IFN-gamma to prime macrophages to respond to trigger signals of LPS and develop cytotoxicity increased with time of incubation, the highest response being observed after 24 h. There was a dose-dependent relationship between the concentrations of both IFN-gamma and LPS used to activate macrophages and the number of dead trophozoites. These data suggest that macrophages are important in host defense against amoebiasis. PMID:2899056

  10. In vitro and in vivo assessment of the anti-malarial activity of Caesalpinia pluviosa

    PubMed Central

    2011-01-01

    Background To overcome the problem of increasing drug resistance, traditional medicines are an important source for potential new anti-malarials. Caesalpinia pluviosa, commonly named "sibipiruna", originates from Brazil and possess multiple therapeutic properties, including anti-malarial activity. Methods Crude extract (CE) was obtained from stem bark by purification using different solvents, resulting in seven fractions. An MTT assay was performed to evaluate cytotoxicity in MCF-7 cells. The CE and its fractions were tested in vitro against chloroquine-sensitive (3D7) and -resistant (S20) strains of Plasmodium falciparum and in vivo in Plasmodium chabaudi-infected mice. In vitro interaction with artesunate and the active C. pluviosa fractions was assessed, and mass spectrometry analyses were conducted. Results At non-toxic concentrations, the 100% ethanolic (F4) and 50% methanolic (F5) fractions possessed significant anti-malarial activity against both 3D7 and S20 strains. Drug interaction assays with artesunate showed a synergistic interaction with the F4. Four days of treatment with this fraction significantly inhibited parasitaemia in mice in a dose-dependent manner. Mass spectrometry analyses revealed the presence of an ion corresponding to m/z 303.0450, suggesting the presence of quercetin. However, a second set of analyses, with a quercetin standard, showed distinct ions of m/z 137 and 153. Conclusions The findings show that the F4 fraction of C. pluviosa exhibits anti-malarial activity in vitro at non-toxic concentrations, which was potentiated in the presence of artesunate. Moreover, this anti-malarial activity was also sustained in vivo after treatment of infected mice. Finally, mass spectrometry analyses suggest that a new compound, most likely an isomer of quercetin, is responsible for the anti-malarial activity of the F4. PMID:21535894

  11. Anti-Helicobacter pylori activities of Chenopodium ambrosioides L. in vitro and in vivo

    PubMed Central

    Ye, Hui; Liu, Yu; Li, Ning; Yu, Jing; Cheng, Hong; Li, Jiang; Zhang, Xue-Zhi

    2015-01-01

    AIM: To investigate the bactericidal effects of Chenopodium ambrosioides L. (CAL) against Helicobacter pylori (H. pylori) both in vitro and in vivo. METHODS: For in vitro experiments, the inhibitory activity of CAL was tested using an agar dilution method; H. pylori strain NCTC11637 was incubated on Columbia blood agar plates containing serial concentrations of CAL. The minimal inhibitory concentration (MIC) was determined by the absence of H. pylori colonies on the agar plate. Time-kill curves were used to evaluate bactericidal activity; the average number of colonies was calculated at 0, 2, 8 and 24 h after liquid incubation with concentrations of CAL at 0.5, 1, and 2 × MIC. For in vivo experiments, H. pylori-infected mice were randomly divided into CAL, triple therapy (lansoprazole, metronidazole, and clarithromycin), blank control, or H. pylori control groups. The eradication ratios were determined by positive findings from rapid urease tests (RUTs) and by histopathology. RESULTS: In vitro, the MIC of CAL against H. pylori was 16 mg/L. The time-kill curves showed a stable and persistent decreasing tendency with increasing CAL concentration, and the intensity of the bactericidal effect was proportional to dose; the 1 and 2 × MIC completely inhibited the growth of H. pylori at 24 h. In vivo, the eradication ratios in the CAL group were 60% (6/10) by RUT and 50% (5/10) by histopathology. Ratios in the triple therapy group were both 70% (7/10), and there was no difference between the CAL and triple therapy groups. Histopathologic evaluation revealed massive bacterial colonization on the surface of gastric mucosa and slight infiltration of mononuclear cells after inoculation with H. pylori, but no obvious inflammation or other pathologic changes in gastric mucosa of mice from CAL and triple therapy groups. CONCLUSION: CAL demonstrates effective bactericidal activity against H. pylori both in vitro and in vivo. PMID:25892867

  12. In Vitro Antifungal Activities of the New Triazole UR-9825 against Clinically Important Filamentous Fungi

    PubMed Central

    Capilla, Javier; Ortoneda, Montserrat; Pastor, Francisco Javier; Guarro, Josep

    2001-01-01

    We used a modified reference microdilution method (the M-38P method) to evaluate the in vitro activities of the new triazole UR-9825 in comparison with those of amphotericin B against 77 strains of opportunistic filamentous fungi. UR-9825 was clearly more active than amphotericin B against all fungi except Fusarium solani and Scytalidium spp. Notably, UR-9825 had low MICs for Aspergillus fumigatus and Paecilomyces lilacinus (MICs at which 90% of isolates are inhibited, 0.125 ?g/ml for both species). PMID:11502542

  13. A humanized anti-M2 scFv shows protective in vitro activity against influenza

    SciTech Connect

    Bradbury, Andrew M; Velappan, Nileena; Schmidt, Jurgen G

    2008-01-01

    M2 is one of the most conserved influenza proteins, and has been widely prospected as a potential universal vaccine target, with protection predominantly mediated by antibodies. In this paper we describe the creation of a humanized single chain Fv from 14C2, a potent monoclonal antibody against M2. We show that the humanized scFv demonstrates similar activity to the parental mAb: it is able to recognize M2 in its native context on cell surfaces and is able to show protective in vitro activity against influenza, and so represents a potential lead antibody candidate for universal prophylactic or therapeutic intervention in influenza.

  14. In Vitro Selection for Small-Molecule-Triggered Strand Displacement and Riboswitch Activity.

    PubMed

    Martini, Laura; Meyer, Adam J; Ellefson, Jared W; Milligan, John N; Forlin, Michele; Ellington, Andrew D; Mansy, Sheref S

    2015-10-16

    An in vitro selection method for ligand-responsive RNA sensors was developed that exploited strand displacement reactions. The RNA library was based on the thiamine pyrophosphate (TPP) riboswitch, and RNA sequences capable of hybridizing to a target duplex DNA in a TPP regulated manner were identified. After three rounds of selection, RNA molecules that mediated a strand exchange reaction upon TPP binding were enriched. The enriched sequences also showed riboswitch activity. Our results demonstrated that small-molecule-responsive nucleic acid sensors can be selected to control the activity of target nucleic acid circuitry. PMID:25978303

  15. Role of paramagnetic polyconjugated clusters in lignin antioxidant activity (in vitro)

    NASA Astrophysics Data System (ADS)

    Dizhbite, T.; Ponomarenko, J.; Andersone, A.; Dobele, G.; Lauberts, M.; Krasilnikova, J.; Mironova-Ulmane, N.; Telysheva, G.

    2012-08-01

    Using physico-chemical methods (EPR, SEC, Py-GC/MS and UV/VIS spectroscopy) and wet chemical analysis, the characteristics of 6 hardwood lignins in terms of functionality, molecular weight and composition of lignin substructures were determined and considered together with the results of DPPH•, ABTS•+ and O2•- antioxidant assays with the aim to understand the relationships governing antioxidant properties of lignin. The strong positive linear correlation between lignin antioxidant capacity in the three assays used and the extent of conjugation of paramagnetic polyconjugated clusters in lignin macromolecules was found. The biological activity of the most active alkaline lignins was assessed by in vitro experiment with human blood.

  16. In Vitro Antifungal Activities of Isavuconazole and Comparators against Rare Yeast Pathogens ?

    PubMed Central

    Guinea, Jesús; Recio, Sandra; Escribano, Pilar; Peláez, Teresa; Gama, Beatriz; Bouza, Emilio

    2010-01-01

    We compared the in vitro activities of isavuconazole, posaconazole, voriconazole, and fluconazole against Dipodascus capitatus (n = 21), Saccharomyces cerevisiae (n = 20), Rhodotorula mucilaginosa (n = 18), and Trichosporon spp. (n = 15). The MIC50s, MIC90s, and MIC ranges (in ?g/ml) obtained using the CLSI M27-A3 procedure were as follows: isavuconazole, 0.125, 0.5, and ?0.015 to 2; posaconazole, 0.5, 2, and ?0.015 to >16; voriconazole, 0.125, 2, and ?0.015 to 8; and fluconazole, 4, >128, and ?0.125 to >128. Isavuconazole showed potent activity against the isolates studied. PMID:20566770

  17. Two activators of in vitro fertilization in mice from licorice.

    PubMed

    Tung, Nguyen Huu; Shoyama, Yukihiro; Wada, Morimasa; Tanaka, Hiromitsu

    2015-11-13

    Systems for artificial insemination have been established in some animals. However, due to limited availability of sperm and oocytes, more effective treatment methodologies are required. Recently, it was demonstrated that the rate of in vitro fertilization (IVF) in mice was improved by adding a water extract of licorice (Glycyrrhiza uralensis), but not glycyrrhizic acid, to the artificial insemination culture medium. In this study, we examined licorice extract for active compounds using bioassay-guided separation. The results indicated that isoliquiritigenin and formononetin were the active molecules in licorice that contributed to the improved rate of IVF. PMID:26392313

  18. Antitumor activity of photodynamic therapy with a chlorin derivative in vitro and in vivo.

    PubMed

    Wang, Lai-Xing; Li, Jian-Wei; Huang, Jian-Yue; Li, Jian-Hong; Zhang, Li-Jun; O'Shea, Donal; Chen, Zhi-Long

    2015-08-01

    Chlorin derivatives are promising photosensitive agents for photodynamic therapy (PDT) of tumors. The aim of the current study is to investigate the PDT therapeutic effects of a novel chlorin-based photosensitizer, meso-tetra[3-(N,N-diethyl)aminomethyl-4-methoxy]phenyl chlorin (TMPC) for gliomas in vitro and in vivo. Physicochemical characteristics of TMPC were recorded by ultraviolet visible spectrophotometer and fluorescence spectrometer. The rate of singlet oxygen generation of TMPC upon photo-excitation was detected by using 1,3-diphenylisobenzofuran (DPBF). The accumulation of TMPC in gliomas U87 MG cells was measured by fluorescence spectrometer. The efficiency of TMPC-PDT in vitro was analyzed by MTT assay and clonogenic assay. The biodistribution and clearance of TMPC were determined by fluorescence measuring. Human gliomas U87 MG tumor-bearing mice model was used to evaluate the antitumor effects of TMPC-PDT. TMPC shows a singlet oxygen generation rate of 0.05 and displays a characteristic long wavelength absorption peak at 653 nm (??=?15,400). The accumulation of TMPC increased with the increase of incubation time. In vitro, PDT using TMPC and laser showed laser dose- and concentration-dependent cytotoxicity to U87 MG cells. In U87 MG tumor-bearing mice, TMPC-PDT significantly reduced the growth of the tumors. Both in vitro and in vivo, TMPC showed little dark toxicity. In vitro and in vivo studies, it found that TMPC has excellent antitumor activities. It suggests that TMPC is a potential photosensitizer of photodynamic therapy for cancer. PMID:25846737

  19. Characterization of human lactoferricin as a potent protein kinase CK2 activator regulated by A-kinase in vitro.

    PubMed

    Maekawa, Toshiro; Fujihara, Michio; Ohtsuki, Kenzo

    2002-01-01

    Lactoferricin (LFcin) hydrolyzed from lactoferrin (LF), a major 80 kDa iron-binding protein in milk and other exocrine secretions, was characterized as a potent activator of protein kinase CK2 (CK2) in vitro. Human LFcin (hLFcin) at 0.5 microg stimulated approx. 5-fold CK2 activity [phosphorylation of 60S acidic ribosomal proteins (P0, P1, P2) and Hsp90 (p98)] in a manner similar to other functional proteins with oligo-Arg clusters, such as salmine A1, sperm histone H2B and HIV-1 Rev. Interestingly, this stimulatory effect of hLFcin was significantly reduced when it was phosphorylated by A-kinase in vitro. These results suggest that (i) hLFcin acts as a potent CK2 activator in vitro; and (ii) the stimulatory effect of hLFcin on CK2 activity is regulated by its phosphorylation by A-kinase in vitro. PMID:11824539

  20. In vitro assays for assessment of androgenic and estrogenic activity of defined mixtures and complex environment samples

    EPA Science Inventory

    Point sources of potentially endocrine active compounds to aquatic environments such as waste water treatment plants, pulp and paper mills, and animal feeding operations invariably contain complex mixtures of chemicals. The current study investigates the use of targeted in vitro ...

  1. Qushi Huayu Decoction Inhibits Hepatic Lipid Accumulation by Activating AMP-Activated Protein Kinase In Vivo and In Vitro

    PubMed Central

    Feng, Qin; Gou, Xiao-jun; Meng, Sheng-xi; Huang, Cheng; Zhang, Yu-quan; Tang, Ya-jun; Wang, Wen-jing; Xu, Lin; Peng, Jing-hua; Hu, Yi-yang

    2013-01-01

    Qushi Huayu Decoction (QHD), a Chinese herbal formula, has been proven effective on alleviating nonalcoholic fatty liver disease (NAFLD) in human and rats. The present study was conducted to investigate whether QHD could inhibit hepatic lipid accumulation by activating AMP-activated protein kinase (AMPK) in vivo and in vitro. Nonalcoholic fatty liver (NAFL) model was duplicated with high-fat diet in rats and with free fatty acid (FFA) in L02 cells. In in vivo experimental condition, QHD significantly decreased the accumulation of fatty droplets in livers, lowered low-density lipoprotein cholesterol (LDL-c), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels in serum. Moreover, QHD supplementation reversed the HFD-induced decrease in the phosphorylation levels of AMPK and acetyl-CoA carboxylase (ACC) and decreased hepatic nuclear protein expression of sterol regulatory element-binding protein-1 (SREBP-1) and carbohydrate-responsive element-binding protein (ChREBP) in the liver. In in vitro, QHD-containing serum decreased the cellular TG content and alleviated the accumulation of fatty droplets in L02 cells. QHD supplementation reversed the FFA-induced decrease in the phosphorylation levels of AMPK and ACC and decreased the hepatic nuclear protein expression of SREBP-1 and ChREBP. Overall results suggest that QHD has significant effect on inhibiting hepatic lipid accumulation via AMPK pathway in vivo and in vitro. PMID:23573117

  2. Specialization of the DNA-Cleaving Activity of a Group I Ribozyme Through In Vitro Evolution

    NASA Technical Reports Server (NTRS)

    Tsang, Joyce; Joyce, Gerald F.

    1996-01-01

    In an earlier study, an in vitro evolution procedure was applied to a large population of variants of the Tetrahymena group 1 ribozyme to obtain individuals with a 10(exp 5)-fold improved ability to cleave a target single-stranded DNA substrate under simulated physiological conditions. The evolved ribozymes also showed a twofold improvement, compared to the wild-type, in their ability to cleave a single-stranded RNA substrate. Here, we report continuation of the in vitro evolution process using a new selection strategy to achieve both enhanced DNA and diminished RNA-cleavage activity. Our strategy combines a positive selection for DNA cleavage with a negative selection against RNA binding. After 36 "generations" of in vitro evolution, the evolved population showed an approx. 100-fold increase in the ratio of DNA to RNA-cleavage activity. Site-directed mutagenesis experiment confirmed the selective advantage of two covarying mutations within the catalytic core of ribozyme that are largely responsible for this modified behavior. The population of ribozymes has now undergone a total of 63 successive generations of evolution, resulting in an average 28 mutations relative to the wild-type that are responsible for the altered phenotype.

  3. Anticoagulants influence the in vitro activity and composition of shock lymph but not its in vivo activity

    PubMed Central

    Deitch, Edwin A.; Qin, Xiaofa; Sheth, Sharvil U.; Tiesi, Gregory; Palange, David; Dong, Wei; Lu, Qi; Xu, DaZhong; Feketeova, Eleonora; Feinman, Rena

    2011-01-01

    Many models of trauma-hemorrhagic shock (T/HS) involve the reinfusion of anticoagulated shed blood. Our recent observation that the anticoagulant heparin induces increased mesenteric lymph lipase activity and consequent in vitro endothelial cell cytotoxicity prompted us to investigate the effect of heparin-induced lipase activity on organ injury in vivo as well as the effects of other anticoagulants on mesenteric lymph bioactivity in vitro and in vivo. To investigate this issue, rats subjected to trauma-hemorrhage had their shed blood anticoagulated with heparin, the synthetic anticoagulant arixtra or citrate. Arixtra, in contrast to heparin, did not increased lymph lipase activity or result in high levels of endothelial cytotoxicity. Yet, the arixtra-treated rats subjected to T/HS still manifested lung injury, neutrophil priming and RBC dysfunction, which was totally abrogated by lymph duct ligation. Furthermore, the injection of T/HS mesenteric lymph, but not sham-shock lymph, collected from the arixtra rats into control mice recreated the pattern of lung injury, PMN priming and RBC dysfunction observed after actual shock. Consistent with these observations, citrate anticoagulated rats subjected to T/HS developed lung injury and the injection of mesenteric lymph from the citrate-anticoagulated T/HS rats into control mice also resulted in lung injury. Based on these results, several conclusions can be drawn. First, heparin-induced increased mesenteric lymph lipase activity is not responsible for the in vivo effects of T/HS mesenteric lymph. Secondly, heparin should be avoided as an anticoagulant when studying the biology or composition of mesenteric lymph due to its ability to cause increases in lymph lipase activity that increase the in vitro cytotoxicity of these lymph samples. PMID:21558984

  4. [In vitro activity of doripenem against strains from pediatric diseases and strains causing purulent meningitis].

    PubMed

    Ohta, Merime; Toba, Shinsuke; Ito, Akinobu; Nakamura, Rio; Tsuji, Masakatsu

    2012-12-01

    This study evaluated the in vitro activity of doripenem (DRPM) against 200 Streptococcus pneumoniae and 197 Haemophilus influenzae from children and adults in 2007, 50 H. influenzae type b in 2006, 20 Listeria monocytogenes in 1990-2005, 23 Neisseria meningitidis in 2007-2009 and 83 Bordetella pertussis in 1989-2003. All strains were isolated from Japanese clinical facilities. We also investigated in vitro activity of other carbapenems (meropenem, imipenem, panipenem, biapenem), cephems (ceftriaxone, cefotaxime), ampicillin and clarithromycin. The all MICs were determined by a broth micro dilution method or an agar dilution method according to CLSI. The MIC90(s) of DRPM against S. pneumoniae and H. influenzae from children were 0.25 microg/mL, 1 microg/mL, respectively, which were similar to strains from adults. These results suggested that antibacterial activity of DRPM is not variable by patient's age. DRPM also showed excellent activities against H. influenzae type b, L. monocytogenes and N. meningitidis, which cause purulent meningitis, and B. pertussis causing whooping cough more than the other carbapenems. DRPM showed superior activities against serious strains of pediatric infection diseases. PMID:23593734

  5. Characterization and antioxidant activity in vitro and in vivo of polysaccharide purified from Rana chensinensis skin.

    PubMed

    Wang, Zhanyong; Zhao, Yuanyuan; Su, Tingting; Zhang, Jing; Wang, Fei

    2015-08-01

    Preliminary characterization and antioxidant activity in vitro and in vivo investigation of the polysaccharide fraction named as RCSP II, which was extracted from Rana chensinensis skin, were performed. Results indicated that RCSP II comprised glucose, galactose, and mannose in a molar ratio of 87.82:2.77:1.54 with a molecular weight of 12.8 kDa. Antioxidant activity assay in vitro showed that RCSP II exhibited 75.2% scavenging activity against 2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) radicals at the concentration of 2500 mg/L and 85.1% against chelated ferrous ion at 4000 mg/L. Antioxidant activity assay in vivo further showed that RCSP II increased the activities of antioxidant enzymes, decreased the levels of malondialodehyde, and enhanced total antioxidant capabilities in livers and sera of d-galactose induced mice. These results suggested that RCSP II could have potential antioxidant applications as medicine or functional food. PMID:25933517

  6. Neuronal medium that supports basic synaptic functions and activity of human neurons in vitro

    PubMed Central

    Bardy, Cedric; van den Hurk, Mark; Eames, Tameji; Marchand, Cynthia; Hernandez, Ruben V.; Kellogg, Mariko; Gorris, Mark; Galet, Ben; Palomares, Vanessa; Brown, Joshua; Bang, Anne G.; Mertens, Jerome; Böhnke, Lena; Boyer, Leah; Simon, Suzanne; Gage, Fred H.

    2015-01-01

    Human cell reprogramming technologies offer access to live human neurons from patients and provide a new alternative for modeling neurological disorders in vitro. Neural electrical activity is the essence of nervous system function in vivo. Therefore, we examined neuronal activity in media widely used to culture neurons. We found that classic basal media, as well as serum, impair action potential generation and synaptic communication. To overcome this problem, we designed a new neuronal medium (BrainPhys basal + serum-free supplements) in which we adjusted the concentrations of inorganic salts, neuroactive amino acids, and energetic substrates. We then tested that this medium adequately supports neuronal activity and survival of human neurons in culture. Long-term exposure to this physiological medium also improved the proportion of neurons that were synaptically active. The medium was designed to culture human neurons but also proved adequate for rodent neurons. The improvement in BrainPhys basal medium to support neurophysiological activity is an important step toward reducing the gap between brain physiological conditions in vivo and neuronal models in vitro. PMID:25870293

  7. Fluorescent and bioluminescent nanoprobes for in vitro and in vivo detection of matrix metalloproteinase activity

    PubMed Central

    Lee, Hawon; Kim, Young-Pil

    2015-01-01

    Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that degrade the extracellular matrix (ECM) and regulate the extracellular microenvironment. Despite the significant role that MMP activity plays in cell-cell and cell-ECM interactions, migration, and differentiation, analyses of MMPs in vitro and in vivo have relied upon their abundance using conventional immunoassays, rather than their enzymatic activities. To resolve this issue, diverse nanoprobes have emerged and proven useful as effective activity-based detection tools. Here, we review the recent advances in luminescent nanoprobes and their applications in in vitro diagnosis and in vivo imaging of MMP activity. Nanoprobes with the purpose of sensing MMP activity consist of recognition and detection units, which include MMP-specific substrates and luminescent (fluorescent or bioluminescent) nanoparticles, respectively. With further research into improvement of the optical performance, it is anticipated that luminescent nanoprobes will have great potential for the study of the functional roles of proteases in cancer biology and nanomedicine. [BMB Reports 2015; 48(6): 313-318] PMID:25817215

  8. In vitro ?-amylase and ?-glucosidase inhibitory effects and cytotoxic activity of Albizia antunesiana extracts

    PubMed Central

    Chipiti, Talent; Ibrahim, Mohammed Auwal; Singh, Moganavelli; Islam, Md. Shahidul

    2015-01-01

    Context: Diabetes mellitus is a chronic disease, and its incidence is tremendously increasing globally. Decreasing postprandial hyperglycemia by retarding glucose absorption through inhibiting carbohydrates digesting enzymes (?-amylase and ?-glucosidase) is one of many approaches used for the management of this disease. Objectives: The leaf and root aqueous and ethanol extracts of Albizia antunesiana were investigated for ?-amylase and ?-glucosidase inhibitory and cytotoxic activity in vitro. Materials and Methods: The ?-amylase and ?-glucosidase activities were measured in the presence of aqueous and ethanol extracts of the plant parts using starch and p-nitrophenyl-D-glucopyranoside as substrates respectively. Furthermore, cytotoxic effects of the extracts were investigated on HEK (human embryonic kidney) 293 cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay. Results: The results showed that ethanolic root extract of A. antunesiana had mild ?-amylase and strong ?-glucosidase inhibitory activity with half-maximal inhibitory concentration values of 30.68 and 4.35 µg/mL, respectively. The aqueous root extract showed mild ?-glucosidase but no ?-amylase inhibitory activity. Cytotoxicity studies on the extracts using the MTT assay revealed that the ethanolic (leaf and root) extracts were relatively nontoxic at tested concentrations on the HEK 293 cell lines. However, the aqueous extracts (leaf and root) were cytotoxic at concentrations above 50 µg/mL. Conclusion: Data from this study suggest that the ethanolic root extract of A. antunesiana possess in vitro ?-amylase and ?-glucosidase inhibitory activities and are not cytotoxic at least in an in vitro condition.

  9. In vitro antioxidant activities of low-molecular-weight polysaccharides with various functional groups.

    PubMed

    Chen, Szu Kai; Tsai, Min Lang; Huang, Jin Ru; Chen, Rong Huei

    2009-04-01

    The objectives of this study were to evaluate the effect of different functional groups of sulfate, amine, and hydroxyl and/or their ionized groups on in vitro antioxidant capacities of low-molecular-weight polysaccharides (LMPS) prepared from agar (LMAG), chitosan (LMCH), and starch (LMST), respectively, and to elucidate their structure-activity relationship. Ascorbic acid and ethylenediaminetetraacetic acid (EDTA) were used as positive controls. The in vitro antioxidant capacities of LMAG and LMCH were higher than that of LMST in the DPPH radical, superoxide radical, hydrogen peroxide, and nitric oxide radical scavenging and ferrous metal-chelating capacities. The different scavenging capacities may be due to the combined effects of the different sizes of the electron-cloud density and the different accessibility between free radical and LMPS, which, in turn, depends upon the different hydrophobicities of the constituent sugars. PMID:19256513

  10. Bio-active nanoemulsions enriched with gold nanoparticle, marigold extracts and lipoic acid: In vitro investigations.

    PubMed

    Guler, Emine; Barlas, F Baris; Yavuz, Murat; Demir, Bilal; Gumus, Z Pinar; Baspinar, Yucel; Coskunol, Hakan; Timur, Suna

    2014-09-01

    A novel and efficient approach for the preparation of enriched herbal formulations was described and their potential applications including wound healing and antioxidant activity (cell based and cell free) were investigated via in vitro cell culture studies. Nigella sativa oil was enriched with Calendula officinalis extract and lipoic acid capped gold nanoparticles (AuNP-LA) using nanoemulsion systems. The combination of these bio-active compounds was used to design oil in water (O/W) and water in oil (W/O) emulsions. The resulted emulsions were characterized by particle size measurements. The phenolic content of each nanoemulsion was examined by using both colorimetric assay and chromatographic analyses. Two different methods containing cell free chemical assay (1-diphenyl-2-picrylhydrazyl method) and cell based antioxidant activity test were used to evaluate the antioxidant capacities. In order to investigate the bio-activities of the herbal formulations, in vitro cell culture experiments, including cytotoxicity, scratch assay, antioxidant activity and cell proliferation were carried out using Vero cell line as a model cell line. Furthermore, to monitor localization of the nanoemulsions after application of the cell culture, the cell images were monitored via fluorescence microscope after FITC labeling. All data confirmed that the enriched N. sativa formulations exhibited better antioxidant and wound healing activity than N. sativa emulsion without any enrichment. In conclusion, the incorporation of AuNP-LA and C. officinalis extract into the N. sativa emulsions significantly increased the bio-activities. The present work may support further studies about using the other bio-active agents for the enrichment of herbal preparations to strengthen their activities. PMID:25009101

  11. Effects of Guanxinning injection on rat cytochrome P450 isoforms activities in vivo and in vitro.

    PubMed

    Yu, Yue; Liu, Yan; Li, Qian; Sun, Jiahui; Lin, Haiou; Liu, Gaofeng

    2015-01-01

    1. We aimed to investigate the regulatory effects of Guanxinning injection (GXNI) on activities of cytochrome P1A2 (CYP1A2), CYP2C11, CYP2D1 and CYP3A1/2 by probe drugs in rats in vivo and in vitro. 2. GXNI-treated and blank control groups were administered GXNI and physiological saline by caudal vein for 14 days consecutively, then they were given the probe drugs of caffeine (10?mg/kg), tolbutamide (10?mg/kg), metoprolol (20?mg/kg) and dapsone (10?mg/kg) by intraperitoneal injection. The blood samples were collected at different times for ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis. Changes of the pharmacokinetics parameters between the GXNI-treated and the blank control groups were used to evaluate the effects of GXNI on the four CYP450 isoforms in rats in vivo. After blood collection, the livers of rats were taken and made microsomes for in vitro tests. The relevant metabolites of phenacetin, tolbutamide, dextromethorphan and testosterone were analyzed quantitatively by high-performance liquid chromatography (HPLC) after microsome incubation. The statistical differences between the two groups were observed to detect the effects of GXNI on the four CYP450 isoforms in rats in vitro. 3. The in vivo and in vitro results demonstrated that GXNI could induce CYP1A2 activity in rats, but had no significant effects on CYP2C11, CYP2D1 and CYP3A1/2. PMID:25495039

  12. Antiplasmodial and cytotoxic activity of coumarin derivatives from dried roots of Angelica gigas Nakai in vitro.

    PubMed

    Moon, Hyung-In; Lee, Jai-Heon; Lee, Young-Choon; Kim, Kyoung-Sook

    2011-12-01

    The butanol-soluble fraction of the dried root of Angelica gigas exhibited significant protection against chloroquine-sensitive strains of Plasmodium falciparum using the parasite lactate dehydrogenase assay method. Using antiplasmodial activity-guided fractionation, five coumarins, marmesinin (1), nodakenin (2), skimmin (3), apiosylskimmin (4), and magnolioside (5), were isolated and evaluated for in vitro antiplasmodial activity, as well as for their cytotoxic potential on SK-OV-3 cancer cell lines. Compounds 1 and 5 showed notable growth inhibitory activity against chloroquine-sensitive strains of P. falciparum with IC(50) values of 5.3 and 8.2 ?M. The compounds showed no significant cytotoxicity (IC(50) > 100 ?M) toward the SK-OV-3 cancer cell line. This is the first report on the antiplasmodial activity of these coumarin derivatives from the dried root of A. gigas. PMID:21428713

  13. [In vitro antifungal activity of rilopirox, a new hydroxypyridone antimycotic agent].

    PubMed

    Harada, I; Mitsui, K; Uchida, K; Yamaguchi, H

    1997-02-01

    In vitro antifungal activities of rilopirox (RIL), a new topical hydroxypyridone antifungal agent, were studied against 7 species (31 strains) of yeast-like fungi and 15 species (17 strains) of mycerial fungi from stock cultures of a wide range of medically important fungi. Tests were carried out by the liquid dilution method using Neopeptone dextrose broth with ciclopirox olamine (CPO) and oxiconazole nitrate (OCZ) as reference drugs. RIL exhibited a broad spectrum of antifungal activities; the MIC of RIL against yeasts were about 1 microgram/ml, those against other fungi were 0.5-4 micrograms/ml. Antifungal activities were similar to CPO, and compare to OCZ, RIL showed characteristically little differences in its activities against different species or strains of target organisms. PMID:9100079

  14. In vitro antiprotozoal and cytotoxic activity of ethnopharmacologically selected guinean plants.

    PubMed

    Traore, Mohammed Sahar; Diane, Sere; Diallo, Mamadou Saliou Telly; Balde, Elhadj Saďdou; Balde, Mamadou Aliou; Camara, Aďssata; Diallo, Abdoulaye; Keita, Abdoulaye; Cos, Paul; Maes, Louis; Pieters, Luc; Balde, Aliou Mamadou

    2014-10-01

    Based on an ethnobotanical survey, 41 Guinean plant species widely used in the traditional treatment of fever and/or malaria were collected. From these, 74 polar and apolar extracts were prepared and tested for their in vitro antiprotozoal activity along with their cytotoxicity on MRC-5 cells. A potent activity (IC50 activity (IC50 8.1?µg/mL) against Leishmania infantum. The selectivity index of the active samples varied from 0.08 to >?100. These results may validate at least in part the traditional use of some of the plant species. PMID:25180493

  15. Conditions Required for the Rapid Activation In Vitro of the Chloroplast Fructose-1,6-bisphosphatase.

    PubMed

    Rosa, L; Whatley, F R

    1984-05-01

    Conditions required for the reductive activation of purified, spinach chloroplast fructose-1,6-bisphosphatase (EC 3.1.3.11) have been determined in vitro. Full reductive activation was observed only when fructose-1,6-bisphosphate and Mg(2+) were present at the same time as the reducing agent (dithiothreitol). Reduction in the absence either of fructose-1,6-bisphosphate or of Mg(2+) slowly and irreversibly inactivated the enzyme. The concentration of fructose-1,6-bisphosphate that must be present during reduction for maximum activation depends upon the divalent cation present: it is highest with Mg(2+), lower with Ca(2+), and lowest when both Mg(2+) and Ca(2+) are present. A scheme for the reductive activation and inactivation of the enzyme is presented. PMID:16663557

  16. Composition and in vitro anticancer activities of the leaf essential oil of Neolitsea variabillima from Taiwan.

    PubMed

    Su, Yu-Chang; Hsu, Kuan-Ping; Wang, Eugene I-Chen; Ho, Chen-Lung

    2013-04-01

    This study investigated the chemical composition and in vitro anticancer activities of the essential oil isolated from the leaf of Neolitsea variabillima. The essential oil was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. Sixty-seven compounds were identified, representing 100% of the oil. The main components identified were trans-beta-ocimene (13.4%), alpha-cadinol (10.5%), terpinen-4-ol (9.3%), tau-cadinol (9.2%), beta-caryophyllene (8.8%), and sabinene (6.7%). The anticancer activities of oil were evaluated. The results showed that the oil exhibited cytotoxic activity against human oral, liver, lung, colon, melanoma, and leukemic cancer cells. The presence of beta-caryophyllene, tau-cadinol, and alpha-cadinol significantly contributed to the anticancer activities of N. variabillima leaf oil. PMID:23738472

  17. Stimulating activity of Chinese medicinal herbs on human lymphocytes in vitro.

    PubMed

    Shan, B E; Yoshida, Y; Sugiura, T; Yamashita, U

    1999-03-01

    The effects of eight kinds of Chinese medicinal herbs (CMH) on human lymphocytes was studied in vitro. The extract of Cinnamomum cassia presl markedly stimulated human lymphocytes to proliferate. Codonopsis pilosula, Oldenlandia diffusa and Rhizoma typhonii weakly stimulated. These extracts enhanced cytotoxic T-lymphocyte (CTL) activity, but failed to enhance natural killer (NK)-cell activity. The extracts of these CMHs have stimulatory effect on immunoglobulin (Ig) production by B-cells and interleukin(IL)-1 production by monocytes. These activities of Cinnamomun cassia presl extract are associated with glycoproteins, whose molecular weight was about 100 KDa. These results suggest that CMH extracts have a stimulating activity on human lymphocytes and these abilities could be used clinically for the treatment of diseases such as cancer. PMID:10348365

  18. Medicinal activities of the leaves of Musa sapientum var. sylvesteris in vitro

    PubMed Central

    Sahaa, Repon Kumer; Acharyaa, Srijan; Shovon, Syed Sohidul Haque; Royb, Priyanka

    2013-01-01

    Objective This study is to investigate the medicinal value of methanolic extract of the leaves of Musa sapientum var. sylvesteris in Bangladesh. Methods Several biochemical assays, thin layer chormatogarphy and ultra-violet spectroscopy were used to detect the presence of various types of compounds in this extract. Antioxidant effects were measured by DPPH scavenging assay, total reducing assay and hydrogen peroxide scavenging assay. Receptor binding activities and hydrogen peroxide induced hemolysis assay were performed by hemagglutination assay and hemolysis assay using erythrocytes. Disk diffusion assay was performed to show the antibacterial effect of the extract. Results Methanolic extract of the leaves showed antioxidant and antibacterial activity in vitro. The extract showed hemaglutination inhibition activities and hydrogen peroxide induced hemolysis inhibition activity of human red blood cells. Conclusion Musa sapientum var. sylvesteris can be an useful medicinal plant. PMID:23730561

  19. Evaluation of the in vitro activity of flumequine against field isolates of Brachyspira hyodysenteriae.

    PubMed

    Aller-Morán, Luis Miguel; Martínez-Lobo, Francisco Javier; Rubio, Pedro; Carvajal, Ana

    2015-12-01

    Flumequine is a quinolone derivative used in veterinary medicine to treat enteric infections, mainly those caused by Gram negative bacteria and also some Gram positive. Some recent reports by field practitioners have suggested that its use in swine dysentery outbreaks can minimize the impact of this disease. This study aims to evaluate the in vitro anti-Brachyspira hyodysenteriae activity of flumequine. Forty eight field isolates of the bacterium were evaluated using a microdilution test. The lack of colon bioavailability studies of flumequine in pigs makes it difficult to establish the true efficacy of this antibiotic for swine dysentery control. Nonetheless, the relatively high values of MIC50 (50?g/mL) and MBC50 (50?g/mL) obtained suggest poor activity against B. hyodysenteriae. Flumequine activity in swine dysentery outbreaks could be related to its activity against other bacteria, different from B. hyodysenteriae, engaged in swine dysentery pathogenesis. PMID:26679795

  20. In vitro and in vivo activities of the nitroimidazole TBA-354 against Mycobacterium tuberculosis.

    PubMed

    Upton, A M; Cho, S; Yang, T J; Kim, Y; Wang, Y; Lu, Y; Wang, B; Xu, J; Mdluli, K; Ma, Z; Franzblau, S G

    2015-01-01

    Nitroimidazoles are a promising new class of antitubercular agents. The nitroimidazo-oxazole delamanid (OPC-67683, Deltyba) is in phase III trials for the treatment of multidrug-resistant tuberculosis, while the nitroimidazo-oxazine PA-824 is entering phase III for drug-sensitive and drug-resistant tuberculosis. TBA-354 (SN31354[(S)-2-nitro-6-((6-(4-trifluoromethoxy)phenyl)pyridine-3-yl)methoxy)-6,7-dihydro-5H-imidazo[2,1-b][1,3]oxazine]) is a pyridine-containing biaryl compound with exceptional efficacy against chronic murine tuberculosis and favorable bioavailability in preliminary rodent studies. It was selected as a potential next-generation antituberculosis nitroimidazole following an extensive medicinal chemistry effort. Here, we further evaluate the pharmacokinetic properties and activity of TBA-354 against Mycobacterium tuberculosis. TBA-354 is narrow spectrum and bactericidal in vitro against replicating and nonreplicating Mycobacterium tuberculosis, with potency similar to that of delamanid and greater than that of PA-824. The addition of serum protein or albumin does not significantly alter this activity. TBA-354 maintains activity against Mycobacterium tuberculosis H37Rv isogenic monoresistant strains and clinical drug-sensitive and drug-resistant isolates. Spontaneous resistant mutants appear at a frequency of 3 × 10(-7). In vitro studies and in vivo studies in mice confirm that TBA-354 has high bioavailability and a long elimination half-life. In vitro studies suggest a low risk of drug-drug interactions. Low-dose aerosol infection models of acute and chronic murine tuberculosis reveal time- and dose-dependent in vivo bactericidal activity that is at least as potent as that of delamanid and more potent than that of PA-824. Its superior potency and pharmacokinetic profile that predicts suitability for once-daily oral dosing suggest that TBA-354 be studied further for its potential as a next-generation nitroimidazole. PMID:25331696

  1. Characterization of the potent in vitro and in vivo antimalarial activities of ionophore compounds.

    PubMed Central

    Gumila, C; Ancelin, M L; Delort, A M; Jeminet, G; Vial, H J

    1997-01-01

    Large-scale in vitro screening of different types of ionophores previously pinpointed nine compounds that were very active and selective in vitro against Plasmodium falciparum; their in vitro and in vivo antimalarial effects were further studied. Addition of the ionophores to synchronized P. falciparum suspensions revealed that all P. falciparum stages were sensitive to the drugs. However, the schizont stages were three- to ninefold more sensitive, and 12 h was required for complete parasite clearance. Pretreatment of healthy erythrocytes with toxic doses of ionophores for 24 to 48 h showed that the activity was not due to an irreversible effect on the host erythrocyte. No preferential ionophore adsorption in infected or uninfected erythrocytes occurred. On the other hand, ionophore molecules strongly bound to serum proteins since increasing the serum concentration from 2 to 50% led to almost a 25-fold parallel increase in the ionophore 50% inhibitory concentration. Mice infected with the malaria parasites Plasmodium vinckei petteri or Plasmodium chabaudi were successfully treated with eight ionophores in a 4-day suppressive test. The 50% effective dose after intraperitoneal administration ranged from 0.4 to 4.1 mg/kg of body weight, and the therapeutic indices were about 5 for all ionophores except monensin A methyl ether, 5-bromo lasalocid A, and gramicidin D, whose therapeutic indices were 12, 18, and 344, respectively. These three compounds were found to be curative, with no recrudescence. Gramicidin D, which presented impressive antimalarial activity, requires parenteral administration, while 5-bromo lasalocid A has the major advantage of being active after oral administration. Overall, the acceptable levels of toxicity and the good in vivo therapeutic indices in the rodent model highlight the interesting potential of these ionophores for the treatment of malaria in higher animals. PMID:9055986

  2. In Vitro and In Vivo Activities of the Nitroimidazole TBA-354 against Mycobacterium tuberculosis

    PubMed Central

    Cho, S.; Yang, T. J.; Kim, Y.; Wang, Y.; Lu, Y.; Wang, B.; Xu, J.; Mdluli, K.; Ma, Z.; Franzblau, S. G.

    2014-01-01

    Nitroimidazoles are a promising new class of antitubercular agents. The nitroimidazo-oxazole delamanid (OPC-67683, Deltyba) is in phase III trials for the treatment of multidrug-resistant tuberculosis, while the nitroimidazo-oxazine PA-824 is entering phase III for drug-sensitive and drug-resistant tuberculosis. TBA-354 (SN31354[(S)-2-nitro-6-((6-(4-trifluoromethoxy)phenyl)pyridine-3-yl)methoxy)-6,7-dihydro-5H-imidazo[2,1-b][1,3]oxazine]) is a pyridine-containing biaryl compound with exceptional efficacy against chronic murine tuberculosis and favorable bioavailability in preliminary rodent studies. It was selected as a potential next-generation antituberculosis nitroimidazole following an extensive medicinal chemistry effort. Here, we further evaluate the pharmacokinetic properties and activity of TBA-354 against Mycobacterium tuberculosis. TBA-354 is narrow spectrum and bactericidal in vitro against replicating and nonreplicating Mycobacterium tuberculosis, with potency similar to that of delamanid and greater than that of PA-824. The addition of serum protein or albumin does not significantly alter this activity. TBA-354 maintains activity against Mycobacterium tuberculosis H37Rv isogenic monoresistant strains and clinical drug-sensitive and drug-resistant isolates. Spontaneous resistant mutants appear at a frequency of 3 × 10?7. In vitro studies and in vivo studies in mice confirm that TBA-354 has high bioavailability and a long elimination half-life. In vitro studies suggest a low risk of drug-drug interactions. Low-dose aerosol infection models of acute and chronic murine tuberculosis reveal time- and dose-dependent in vivo bactericidal activity that is at least as potent as that of delamanid and more potent than that of PA-824. Its superior potency and pharmacokinetic profile that predicts suitability for once-daily oral dosing suggest that TBA-354 be studied further for its potential as a next-generation nitroimidazole. PMID:25331696

  3. In vitro activities of amphotericin B deoxycholate and liposomal amphotericin B against 604 clinical yeast isolates.

    PubMed

    Montagna, Maria Teresa; Lovero, Grazia; Coretti, Caterina; De Giglio, Osvalda; Martinelli, Domenico; Bedini, Andrea; Delia, Mario; Rosato, Antonio; Codeluppi, Mauro; Caggiano, Giuseppina

    2014-12-01

    We determined the in vitro antifungal activity of liposomal amphotericin B (L-AmB) against 604 clinical yeast isolates. Amphotericin B deoxycholate (D-AmB) was tested in parallel against all the isolates. Susceptibility testing was performed according to the Clinical and Laboratory Standards Institute (CLSI) M27-A3 method. Overall, L-AmB was highly active against the isolates (mean MIC, 0.42 µg ml(-1); MIC90, 1 µg ml(-1); 97.2?% of MICs were ?1 µg ml(-1)) and comparable to D-AmB (mean MIC, 0.48 µg ml(-1); MIC90, 1 µg ml(-1); 97.3?% of MICs were ?1 µg ml(-1)). The in vitro activity of D-AmB and L-AmB was correlated (R(2)?=?0.61; exp(b), 2.3; 95?% CI, 2.19-2.44, P<0.001). Candida albicans (mean MICs of D-AmB and L-AmB, 0.39 µg ml(-1) and 0.31 µg ml(-1), respectively) and Candida parapsilosis (mean MICs of D-AmB and L-AmB, 0.38 µg ml(-1) and 0.35 µg ml(-1), respectively) were the species most susceptible to the agents tested, while Candida krusei (currently named Issatchenkia orientalis) (mean MICs of D-AmB and L-AmB, 1.27 µg ml(-1) and 1.13 µg ml(-1), respectively) was the least susceptible. The excellent in vitro activity of L-AmB may have important implications for empirical treatment approaches and support its role in treatment of a wide range of invasive infections due to yeasts. PMID:25210203

  4. In vitro activities of amphotericin B deoxycholate and liposomal amphotericin B against 604 clinical yeast isolates

    PubMed Central

    Lovero, Grazia; Coretti, Caterina; De Giglio, Osvalda; Martinelli, Domenico; Bedini, Andrea; Delia, Mario; Rosato, Antonio; Codeluppi, Mauro; Caggiano, Giuseppina

    2014-01-01

    We determined the in vitro antifungal activity of liposomal amphotericin B (L-AmB) against 604 clinical yeast isolates. Amphotericin B deoxycholate (D-AmB) was tested in parallel against all the isolates. Susceptibility testing was performed according to the Clinical and Laboratory Standards Institute (CLSI) M27-A3 method. Overall, L-AmB was highly active against the isolates (mean MIC, 0.42 µg ml?1; MIC90, 1 µg ml?1; 97.2?% of MICs were ?1 µg ml?1) and comparable to D-AmB (mean MIC, 0.48 µg ml?1; MIC90, 1 µg ml?1; 97.3?% of MICs were ?1 µg ml?1). The in vitro activity of D-AmB and L-AmB was correlated (R2?=?0.61; exp(b), 2.3; 95?% CI, 2.19–2.44, P<0.001). Candida albicans (mean MICs of D-AmB and L-AmB, 0.39 µg ml?1 and 0.31 µg ml?1, respectively) and Candida parapsilosis (mean MICs of D-AmB and L-AmB, 0.38 µg ml?1 and 0.35 µg ml?1, respectively) were the species most susceptible to the agents tested, while Candida krusei (currently named Issatchenkia orientalis) (mean MICs of D-AmB and L-AmB, 1.27 µg ml?1 and 1.13 µg ml?1, respectively) was the least susceptible. The excellent in vitro activity of L-AmB may have important implications for empirical treatment approaches and support its role in treatment of a wide range of invasive infections due to yeasts. PMID:25210203

  5. SPECT/CT localization of oral radioiodine activity: a retrospective study and in-vitro assessment

    PubMed Central

    Burlison, Jared S.; Hartshorne, Michael F.; Voda, Alan M.; Cocks, Franklin H.

    2013-01-01

    Purpose We sought to further localize radioiodine activity in the mouth on post-thyroid cancer therapy imaging using single-photon emission computed tomography/computed tomography (SPECT/CT). Materials and methods We retrospectively reviewed all patients (58) who underwent thyroid cancer therapy with iodine-131 (131I) at our institution from August 2009 to March 2011 whose post-therapy radioiodine imaging included neck SPECT/CT. A small group (six) of diagnostic 123I scans including SPECT/CT was also reviewed. Separately, we performed in-vitro 131I (sodium iodide) binding assays with amalgam and Argenco HP 77 (77% dental gold alloy) as proof of principle for these interactions. Results Of the 58 post-therapy patients, 45 (78%) had undergone metallic dental restorations, and of them 41 (91%) demonstrated oral 131I activity localizing preferentially to those restorations. It was observed that radioiodine also localized to other dental restorations and to orthodontic hardware. Gum-line activity in edentulous patients suggests radioiodine interaction with denture adhesive. In vitro, dental amalgam and Argenco HP 77 bound 131I in a time-dependent manner over 1–16 days of exposure. Despite subsequent washings with normal saline, significant 131I activity (maximally 12% for amalgam and 68% for Argenco HP 77) was retained by these metals. Subsequent soaking in a saturated solution of potassium iodide partially displaced 131I from amalgam, with near-total displacement of 131I from Argenco HP 77. Conclusion SPECT/CT shows that radioiodine in the oral cavity localizes to metallic dental restorations. Furthermore, in-vitro studies demonstrate partially reversible binding of 131I to common dental metals. PMID:24128897

  6. Potassium humate inhibits complement activation and the production of inflammatory cytokines in vitro

    SciTech Connect

    van Rensburg, C.E.J.; Naude, P.J.

    2009-08-15

    The effects of brown coal derived potassium humate on lymphocyte proliferation, cytokine production and complement activation were investigated in vitro. Potassium humate increased lymphocyte proliferation of phytohaemaglutinin A (PHA) and pokeweed mitogen (PWM) stimulated mononuclear lymphocytes (MNL) in vitro from concentrations of 20 to 80 {mu} g/ml, in a dose dependant manner. On the other hand potassium humate, at 40 {mu} g/ml, significantly inhibited the release of TNF-alpha, IL-1 beta, IL-6 and IL-10 by PHA stimulated MNL. Regarding complement activation it was found that potassium humate inhibits the activation of both the alternative and classical pathways without affecting the stability of the red blood cell membranes. These results indicate that the anti-inflammatory potential of potassium humate could be partially due to the inhibition of pro-inflammatory cytokines responsible for the initiation of these reactions as well as inhibition of complement activation. The increased lymphocyte proliferation observed, might be due to increased IL-2 production as previously been documented.

  7. Improving vagal activity ameliorates cardiac fibrosis induced by angiotensin II: in vivo and in vitro.

    PubMed

    Liu, Jin-Jun; Huang, Ning; Lu, Yi; Zhao, Mei; Yu, Xiao-Jiang; Yang, Yang; Yang, Yong-Hua; Zang, Wei-Jin

    2015-01-01

    Cardiac remodeling is characterized by overactivity of the renin-angiotensin system (RAS) and withdrawal of vagal activity. We hypothesized that improving vagal activity could attenuate cardiac fibrosis induced by angiotensin II (Ang II) in vivo and in vitro. Rats were subjected to abdominal aorta constriction (AAC) with or without pyridostigmine (PYR) (31?mg/kg/d). After 8 weeks, PYR significantly decreased Ang II level, AT1 protein expression, and collagen deposition in cardiac tissue and improved heart rate variability, baroreflex sensitivity and cardiac function, which were abolished by atropine. In vitro, treatment of cardiac fibroblasts (CFs) with Ang II (10(-7)?M) increased cell proliferation, migration, transformation, and secretory properties, which were significantly diminished by acetylcholine (ACh, 10(-6)?M). Subsequently, Ang II significantly increased collagen type I expression as well as metalloproteinase (MMP)-2 expression and activity. Transforming growth factor (TGF)-?1 expression and Smad3 phosphorylation presented a similar trend. Notably, the knockdown of the acetylcholine M2 receptor by siRNA could abolish ACh anti-fibrotic action. These data implicated cholinesterase inhibitor can increase vagal activity and reduce local Ang II level, and ACh inhibit Ang II pro-fibrotic effects. Our findings suggested that the parasympathetic nervous system can serve as a promising target for cardiac remodeling treatment. PMID:26596640

  8. In vitro biological activity of secondary metabolites from Seseli rigidum Waldst. et Kit. (Apiaceae).

    PubMed

    Jakovljevi?, Dragana; Vasi?, Sava; Stankovi?, Milan; ?omi?, Ljiljana; Topuzovi?, Marina

    2015-12-01

    The antioxidant, antimicrobial activity, total phenolic content and flavonoid concentration of Seseli rigidum Waldst. et Kit. were evaluated. Five different extracts of the aboveground plant parts were obtained by extraction with distilled water, methanol, acetone, ethyl acetate and petroleum ether. Total phenols were determined using the Folin-Ciocalteu's reagent, with the highest values obtained in the acetone extract (102.13 mg GAE/g). The concentration of flavonoids, determined by using a spectrophotometric method with aluminum chloride and expressed in terms of rutin equivalent, was also highest in the acetone extracts (291.58 mg RUE/g). The antioxidant activity was determined in vitro using DPPH reagent. The greatest antioxidant activity was expressed in the aqueous extract (46.15 ?g/ml). In vitro antimicrobial activities were determined using a microdilution analysis method; minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) were determined. Methanolic extract had the greatest influence on bacilli (MIC at 0.0391 mg/ml), but the best antimicrobial effect had acetone and ethyl acetate extracts considering their broad impact on bacteria. According to our research, S. rigidum can be regarded as promising candidate for natural plant source with high value of biological compounds. PMID:26616372

  9. An in vitro study into the effect of zinc substituted hydroxyapatite on osteoclast number and activity.

    PubMed

    Shepherd, David V; Kauppinen, Kyösti; Brooks, Roger A; Best, Serena M

    2014-11-01

    Zinc ions have been shown to inhibit osteoclast development and proliferation both in vitro and in vivo. The same inhibiting effect has been observed in vitro when zinc was substituted into tri-calcium phosphate (TCP). Because of the solubility of TCP it is not an ideal candidate for a material to inhibit osteoclast activity in the long term. Hydroxyapatite (HA) is less soluble and so potentially offers a more long-term, sustainable effect. Previous work has shown that zinc can successfully be substituted into HA and still retain phase purity after heat treatment. The study reported here presents the effects of zinc substituted HA on the development and activity of osteoclast-like cells. It was found that increasing zinc substitution levels led to a decrease in the number of these cells present after 21 days. When resorption activity was investigated it was found that an increase in the amount of zinc present in the discs led to a significant decrease in the amount of resorption taking place on the discs. These results provide evidence for the potential of zinc substituted HA as a material to reduce resorptive activity to provide long-term bonding of implant to bone. PMID:24443251

  10. In vitro activity of 23 tea extractions and epigallocatechin gallate against Candida species.

    PubMed

    Chen, Ming; Zhai, Lin; Arendrup, Maiken Cavling

    2015-02-01

    In this study, we investigate the susceptibility of Candida albicans, Candida glabrata, Candida krusei, Candida parapsilosis, Candida tropicalis, and Aspergillus fumigatus using the EUCAST microdilution minimum inhibitory concentration (MIC) method (final tea supernatant concentration range 5.0-0.005 mg/ml) to 23 different teas and tea catechins including epigallocatechin gallate (EGCG) isolated from green tea. All teas exhibited potent in vitro antifungal activity against C. glabrata. Six out of nine green teas and three of eight black teas had an MIC of 0.078 mg/ml, one white tea had an MIC of 0.156 mg/ml, and finally three of five oolong teas had an MIC of 0.156 mg/ml. Three teas exhibited activity against C. albicans (MIC 1.25 mg/ml), one green tea was active against C. parapsilosis (MIC 1.25 mg/ml), but none were effective against C. krusei, C. tropicalis or A. fumigatus at the concentrations tested. The MIC of EGCG was 0.3125 ?g/ml against C. glabrata and 5.0 ?g/ml against C. albicans and C. parapsilosis. The effect was fungicidal against C. glabrata at higher concentrations. In conclusion, EGCG and other yet undefined substances in tea have differential antifungal activity in vitro against C. glabrata, C. albicans and C. parapsilosis. These data indicate that components of tea and EGCG might be useful particularly for the treatment of C. glabrata infections and warrants further investigations. PMID:25605775

  11. In vitro Antioxidant and Antibacterial Activities of Methanol Extract of Kyllinga nemoralis.

    PubMed

    Sindhu, T; Rajamanikandan, S; Srinivasan, P

    2014-03-01

    The present study was designed to evaluate the antioxidant and antibacterial activity of methanol extract of Kyllinga nemoralis. Six different in vitro antioxidant assays including 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical, superoxide anion radical, hydrogen peroxide radical, ferric reducing antioxidant power assay and reducing power were carried out to ensure the scavenging effect of the plant on free radicals. In addition, total antioxidant capacity assay, total phenolic contents, tannins, flavonoids and flavonol contents of the plant were also analysed by the standard protocols. Kyllinga nemoralis exhibited high antioxidant activity on 2,2-diphenyl-1-picrylhydrazyl assay (IC50= 90 ?g/ml), superoxide radical scavenging assay (IC50= 180 ?g/ml) and hydrogen peroxide radical scavenging assay (IC50= 200 ?g/ml), compared with standards. These observations provide comprehensible supporting evidence for the antioxidant potential of the plant extract. Reducing power (IC50= 213.16 ?g/ml) and hydroxyl radical scavenging activity (IC50= 223 ?g/ml) of the plant extract was remarkable. The methanol extract of K. nemoralis exhibited significant antimicrobial activity against Gram-positive human pathogenic bacteria. Standard in vitro antioxidant assays assessed the electron donating ability of the plant extract in scavenging free radicals. The inhibitory effect of the plant extract against bacterial pathogens may be due to the presence of phytochemicals. Thus, the results suggest that Kyllinga nemoralis is a potential source of antioxidants and could serve as the base for drug development. PMID:24843192

  12. In vitro Antioxidant and Antibacterial Activities of Methanol Extract of Kyllinga nemoralis

    PubMed Central

    Sindhu, T.; Rajamanikandan, S.; Srinivasan, P.

    2014-01-01

    The present study was designed to evaluate the antioxidant and antibacterial activity of methanol extract of Kyllinga nemoralis. Six different in vitro antioxidant assays including 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical, superoxide anion radical, hydrogen peroxide radical, ferric reducing antioxidant power assay and reducing power were carried out to ensure the scavenging effect of the plant on free radicals. In addition, total antioxidant capacity assay, total phenolic contents, tannins, flavonoids and flavonol contents of the plant were also analysed by the standard protocols. Kyllinga nemoralis exhibited high antioxidant activity on 2,2-diphenyl-1-picrylhydrazyl assay (IC50= 90 ?g/ml), superoxide radical scavenging assay (IC50= 180 ?g/ml) and hydrogen peroxide radical scavenging assay (IC50= 200 ?g/ml), compared with standards. These observations provide comprehensible supporting evidence for the antioxidant potential of the plant extract. Reducing power (IC50= 213.16 ?g/ml) and hydroxyl radical scavenging activity (IC50= 223 ?g/ml) of the plant extract was remarkable. The methanol extract of K. nemoralis exhibited significant antimicrobial activity against Gram-positive human pathogenic bacteria. Standard in vitro antioxidant assays assessed the electron donating ability of the plant extract in scavenging free radicals. The inhibitory effect of the plant extract against bacterial pathogens may be due to the presence of phytochemicals. Thus, the results suggest that Kyllinga nemoralis is a potential source of antioxidants and could serve as the base for drug development. PMID:24843192

  13. Cholecalciferol synthesized after UV-activation of 7-dehydrocholesterol onto titanium implants inhibits osteoclastogenesis in vitro.

    PubMed

    Satué, María; Ramis, Joana M; Monjo, Marta

    2015-07-01

    UV-activated 7-dehydrocholesterol (7-DHC) has been successfully used as a biocompatible coating for titanium (Ti) implants producing active vitamin D with positive effect on osteoblast differentiation. Since an osseointegrating implant must promote bone formation while delay resorption, here we determine the effect of this coating on the pre-osteoclast cell line RAW 264.7. Moreover, D3 synthesis was optimized by (1) the supplementation with VitE of the 7-DHC coating to reduce 7-DHC oxidation and (2) the addition of an incubation step (48 h at 23°C) after UV-irradiation to favor isomerization. In vitro results with RAW264.7 cells showed no cytotoxic effect of the coatings and a significant decrease of osteoclastogenesis. Indeed, TRAP immunostaining suggested an inhibition of Trap-positive multinucleated cells and the mRNA levels of different phenotypic, fusion, and activity markers were reduced, particularly with 7-DHC:VitE. In conclusion, we demonstrate an improvement of the D3 synthesis from UV-activated 7-DHC when combined with VitE and show that these implants inhibit osteoclastogenesis in vitro. PMID:25369149

  14. Improving vagal activity ameliorates cardiac fibrosis induced by angiotensin II: in vivo and in vitro

    PubMed Central

    Liu, Jin-Jun; Huang, Ning; Lu, Yi; Zhao, Mei; Yu, Xiao-Jiang; Yang, Yang; Yang, Yong-hua; Zang, Wei-Jin

    2015-01-01

    Cardiac remodeling is characterized by overactivity of the renin–angiotensin system (RAS) and withdrawal of vagal activity. We hypothesized that improving vagal activity could attenuate cardiac fibrosis induced by angiotensin II (Ang II) in vivo and in vitro. Rats were subjected to abdominal aorta constriction (AAC) with or without pyridostigmine (PYR) (31?mg/kg/d). After 8 weeks, PYR significantly decreased Ang II level, AT1 protein expression, and collagen deposition in cardiac tissue and improved heart rate variability, baroreflex sensitivity and cardiac function, which were abolished by atropine. In vitro, treatment of cardiac fibroblasts (CFs) with Ang II (10?7?M) increased cell proliferation, migration, transformation, and secretory properties, which were significantly diminished by acetylcholine (ACh, 10?6?M). Subsequently, Ang II significantly increased collagen type I expression as well as metalloproteinase (MMP)-2 expression and activity. Transforming growth factor (TGF)-?1 expression and Smad3 phosphorylation presented a similar trend. Notably, the knockdown of the acetylcholine M2 receptor by siRNA could abolish ACh anti-fibrotic action. These data implicated cholinesterase inhibitor can increase vagal activity and reduce local Ang II level, and ACh inhibit Ang II pro-fibrotic effects. Our findings suggested that the parasympathetic nervous system can serve as a promising target for cardiac remodeling treatment. PMID:26596640

  15. The Arabidopsis thaliana SERK1 Kinase Domain Spontaneously Refolds to an Active State In Vitro

    PubMed Central

    aan den Toorn, Marije; Huijbers, Mieke M. E.; de Vries, Sacco C.; van Mierlo, Carlo P. M.

    2012-01-01

    Auto-phosphorylating kinase activity of plant leucine-rich-repeat receptor-like kinases (LRR-RLK's) needs to be under tight negative control to avoid unscheduled activation. One way to achieve this would be to keep these kinase domains as intrinsically disordered protein (IDP) during synthesis and transport to its final location. Subsequent folding, which may depend on chaperone activity or presence of interaction partners, is then required for full activation of the kinase domain. Bacterially produced SERK1 kinase domain was previously shown to be an active Ser/Thr kinase. SERK1 is predicted to contain a disordered region in kinase domains X and XI. Here, we show that loss of structure of the SERK1 kinase domain during unfolding is intimately linked to loss of activity. Phosphorylation of the SERK1 kinase domain neither changes its structure nor its stability. Unfolded SERK1 kinase has no autophosphorylation activity and upon removal of denaturant about one half of the protein population spontaneously refolds to an active protein in vitro. Thus, neither chaperones nor interaction partners are required during folding of this protein to its catalytically active state. PMID:23236403

  16. Pancreatic lipase inhibitory activity of taraxacum officinale in vitro and in vivo

    PubMed Central

    Zhang, Jian; Kang, Min-Jung; Kim, Myung-Jin; Kim, Mi-Eun; Song, Ji-Hyun; Lee, Young-Min

    2008-01-01

    Obesity has become a worldwide health problem. Orlistat, an inhibitor of pancreatic lipase, is currently approved as an anti-obesity drug. However, gastrointestinal side effects caused by Orlistat may limit its use. In this study the inhibitory activities of dandelion (Taraxacum officinale) against pancreatic lipase in vitro and in vivo were measured to determine its possible use as a natural anti-obesity agent. The inhibitory activities of the 95% ethanol extract of T. officinale and Orlistat were measured using 4-methylumbelliferyl oleate (4-MU oleate) as a substrate at concentrations of 250, 125, 100, 25, 12.5 and 4 µg/ml. To determine pancreatic lipase inhibitory activity in vivo, mice (n=16) were orally administered with corn oil emulsion (5 ml/kg) alone or with the 95% ethanol extract of T. officinale (400 mg/kg) following an overnight fast. Plasma triglyceride levels were measured at 0, 90, 180, and 240 min after treatment and incremental areas under the response curves (AUC) were calculated. The 95% ethanol extract of T. officinale and Orlistat, inhibited, porcine pancreatic lipase activity by 86.3% and 95.7% at a concentration of 250 µg/ml, respectively. T. officinale extract showed dose-dependent inhibition with the IC50 of 78.2 µg/ml. A single oral dose of the extract significantly inhibited increases in plasma triglyceride levels at 90 and 180 min and reduced AUC of plasma triglyceride response curve (p<0.05). The results indicate that T. officinale exhibits inhibitory activities against pancreatic lipase in vitro and in vivo. Further studies to elucidate anti-obesity effects of chronic consumption of T. officinale and to identify the active components responsible for inhibitory activity against pancreatic lipase are necessary. PMID:20016719

  17. In Vitro Activity of RX-P873 against Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter baumannii

    PubMed Central

    Rhomberg, Paul R.; Jones, Ronald N.; Farrell, David J.

    2015-01-01

    RX-P873 is a novel antibiotic from the pyrrolocytosine series which exhibits high binding affinity for the bacterial ribosome and broad-spectrum antibiotic properties. The pyrrolocytosines have shown in vitro activity against multidrug-resistant Gram-negative and Gram-positive strains of bacteria known to cause complicated urinary tract, skin, and lung infections, as well as sepsis. Enterobacteriaceae (657), Pseudomonas aeruginosa (200), and Acinetobacter baumannii (202) isolates from North America and Europe collected in 2012 as part of a worldwide surveillance program were tested in vitro by broth microdilution using Clinical and Laboratory Standards Institute (CLSI) methodology. RX-P873 (MIC90, 0.5 ?g/ml) was >32-fold more active than ceftazidime and inhibited 97.1% and 99.5% of Enterobacteriaceae isolates at MIC values of ?1 and ?4 ?g/ml, respectively. There were only three isolates with an MIC value of >4 ?g/ml (all were indole-positive Protea). RX-P873 (MIC50/90, 2/4 ?g/ml) was highly active against Pseudomonas aeruginosa isolates, including isolates which were nonsusceptible to ceftazidime or meropenem. RX-P873 was 2-fold less active against P. aeruginosa than tobramycin (MIC90, 2 ?g/ml; 91.0% susceptible) and colistin (MIC90, 2 ?g/ml; 99.5% susceptible) and 2-fold more potent than amikacin (MIC90, 8 ?g/ml; 93.5% susceptible) and meropenem (MIC90, 8 ?g/ml; 76.0% susceptible). RX-P873, the most active agent against Acinetobacter baumannii (MIC90, 1 ?g/ml), was 2-fold more active than colistin (MIC90, 2 ?g/ml; 97.0% susceptible) and 4-fold more active than tigecycline (MIC90, 4 ?g/ml). This novel agent merits further exploration of its potential against multidrug-resistant Gram-negative bacteria. PMID:25645834

  18. Activities of Psilostachyin A and Cynaropicrin against Trypanosoma cruzi In Vitro and In Vivo

    PubMed Central

    da Silva, Cristiane França; Batista, Denise da Gama Jaen; De Araújo, Julianna Siciliano; Batista, Marcos Meuser; Lionel, Jessica; de Souza, Elen Mello; Hammer, Erica Ripoll; da Silva, Patricia Bernardino; De Mieri, Maria; Adams, Michael; Zimmermann, Stefanie; Hamburger, Matthias; Brun, Reto; Schühly, Wolfgang

    2013-01-01

    In vitro and in vivo activities against Trypanosoma cruzi were evaluated for two sesquiterpene lactones: psilostachyin A and cynaropicrin. Cynaropicrin had previously been shown to potently inhibit African trypanosomes in vivo, and psilostachyin A had been reported to show in vivo effects against T. cruzi, albeit in another test design. In vitro data showed that cynaropicrin was more effective than psilostachyin A. Ultrastructural alterations induced by cynaropicrin included shedding events, detachment of large portions of the plasma membrane, and vesicular bodies and large vacuoles containing membranous structures, suggestive of parasite autophagy. Acute toxicity studies showed that one of two mice died at a cynaropicrin dose of 400 mg/kg of body weight given intraperitoneally (i.p.). Although no major plasma biochemical alterations could be detected, histopathology demonstrated that the liver was the most affected organ in cynaropicrin-treated animals. Although cynaropicrin was as effective as benznidazole against trypomastigotes in vitro, the treatment (once or twice a day) of T. cruzi-infected mice (up to 50 mg/kg/day cynaropicrin) did not suppress parasitemia or protect against mortality induced by the Y and Colombiana strains. Psilostachyin A (0.5 to 50 mg/kg/day given once a day) was not effective in the acute model of T. cruzi infection (Y strain), reaching 100% animal mortality. Our data demonstrate that although it is very promising against African trypanosomes, cynaropicrin does not show efficacy compared to benznidazole in acute mouse models of T. cruzi infection. PMID:23939901

  19. In Vitro Cestocidal Activity of Thymol on Mesocestoides corti Tetrathyridia and Adult Worms.

    PubMed

    Maggiore, M; Elissondo, M C

    2014-01-01

    Nothing is known about the possible effect of thymol or other compounds of essential oils against the adult worms of cestodes. The aim of the present work was to determine in vitro cestodicidal activity of thymol against Mesocestoides corti adult worms. Moreover, the in vitro effect on tetrathyridia was also demonstrated. Tetrathyridia exposed to different concentrations of thymol showed a concentration and time-dependent effect. At lower concentrations, the main change observed was mainly in morphology, with larvae exhibiting an elongation of the body. When tetrathyridia were exposed to higher concentrations, increased surface alterations and damage were detected. The body appeared elongated and flattened, and a complete loss of morphology and microtriches was observed. Thymol was able to kill M. corti tetrathyridia, since following inoculation of treated parasites in mice no parasites could be recovered. The effect on M. corti adult worms was dose and time-dependent. Changes in motility coincide with the tissue damage were observed at the structural and ultrastructural level. Thymol caused severe damages to both developmental stages analyzed. Damages were more significant in fully segmented worms. The data reported in this paper demonstrate a clear in vitro effect of thymol against M. corti tetrathyridia and adult worms. PMID:25258624

  20. In vitro testing of fungicidal activity of biocides against Aspergillus fumigatus.

    PubMed

    Tortorano, Anna Maria; Viviani, Maria Anna; Biraghi, Emanuela; Rigoni, Anna Lisa; Prigitano, Anna; Grillot, Renée

    2005-10-01

    The activity of biocides against Aspergillus fumigatus is unknown. In the European guidelines to evaluate the fungicidal activity of a biocide, the critical step concerning the preparation of conidial suspensions is cumbersome and time-consuming. The aims of this study were to evaluate a simplified procedure to prepare conidial suspensions to test a biocide in comparison with the recommended one and to investigate the in vitro activity of seven biocides by the suspension neutralization method against A. fumigatus clinical isolates. The proposed simplified procedure proved reproducible, gave the same results and was quicker than that described in the European guidelines. Benzalkonium chloride (0.25 %), glutaraldehyde (1.6 %), polyvinylpyrrolidone iodine (1 % available iodine) and polyester glycol iodine (0.18 % available iodine) showed biocidal activity in activity. In addition, a simplified and reproducible procedure may be used for testing the fungicidal activity of new compounds or combined formulations. In conclusion, the biocides tested, which are commonly used in hospital settings, were shown to display biocidal activity against A. fumigatus and a simplified procedure may be adopted for testing the fungicidal activity of new compounds. PMID:16157549

  1. Cytokine activation of murine macrophages for in vitro killing of Entamoeba histolytica trophozoites.

    PubMed

    Denis, M; Chadee, K

    1989-06-01

    Macrophage-mediated effector mechanisms against the protozoan parasite Entamoeba histolytica were studied. Unstimulated macrophages were inefficient at killing E. histolytica trophozoites in vitro and were killed by the trophozoites. Conversely, immature cells of the mononuclear phagocyte lineage (promonocytes) were shown to display a strong spontaneous amebicidal activity. The acquisition of macrophage amebicidal activity following cytokine treatment was investigated. Gamma interferon, tumor necrosis factor alpha, and macrophage colony-stimulating factor 1, or combinations thereof, were shown to endow murine bone marrow-derived macrophages with significant amebicidal activity. Low doses of gamma interferon and tumor necrosis factor alpha and of gamma interferon and colony-stimulating factor 1 were shown to act synergistically in this phenomenon. This enhancement of amebicidal activity was shown to operate on bone marrow-derived macrophages, elicited peritoneal macrophages, and, to a much lesser extent, spleen macrophages. Although acquisition of amebicidal activity was associated with a strong respiratory burst, the addition of oxygen-free radical scavengers showed that the killing activity was approximately 45% H2O2 dependent. In addition, amebicidal activity by macrophages was shown to be contact dependent and was inhibited by 61% with the protease inhibitor tosyl lysyl chloromethyl ketone. Our results indicate that immunologic production of gamma interferon, tumor necrosis factor alpha, and colony-stimulating factor 1 could be important in the activation of macrophages for host defense against amebiasis and that promonocytes are strong effector cells against virulent amebae. PMID:2542164

  2. In Vitro Activation of Feline Immunodeficiency Virus in Ramified Microglial Cells from Asymptomatically Infected Cats

    PubMed Central

    Hein, Andreas; Martin, Jean-Pierre; Dörries, Rüdiger

    2001-01-01

    Intravenous infection of cats with feline immunodeficiency virus was used as a model system to study activation of virus replication in brain-resident microglial cells in vitro. Virus release by ramified microglial cells isolated from subclinically infected animals was detectable in cell-free tissue culture supernatant only by reverse transcription and nested PCR of gag-specific RNA sequences and not by virion-associated reverse transcriptase activity. In contrast, cocultivation of in vivo-infected microglial cells with mitogen-activated peripheral blood mononuclear cells (PBMC) regularly allows detection of high virus yields in cell-free tissue culture fluid. Besides uptake and multiplication of microglia-derived virus in PBMC, release of virus from microglia is stimulated by cell contact with PBMC. The data suggest that T lymphocytes patrolling the central nervous system could reactivate the semilatent state of lentiviruses in microglial cells in the course of clinically silent central nervous system infection. PMID:11483754

  3. In vitro antifungal activity of dihydroxyacetone against causative agents of dermatomycosis.

    PubMed

    Stopiglia, Cheila Denise Ottonelli; Vieira, Fabiane Jamono; Mondadori, Andressa Grazziotin; Oppe, Tércio Paschke; Scroferneker, Maria Lúcia

    2011-04-01

    Dihydroxyacetone (DHA), a three-carbon sugar, is the browning ingredient in commercial sunless tanning formulations. DHA preparations have been used for more than 50 years and are currently highly popular for producing temporary pigmentation resembling an ultraviolet-induced tan. In this work, the in vitro antifungal activity of dihydroxyacetone was tested against causative agents of dermatomycosis, more specifically against dermatophytes and Candida spp. The antifungal activity was determined by the broth microdilution method according to the Clinical and Laboratory Standards Institute guidelines for yeasts and filamentous fungi. The data obtained show that the fungicidal activity varied from 1.6 to 50 mg ml(-1). DHA seems to be a promising substance for the treatment of dermatomycosis because it has antifungal properties at the same concentration used in artificial suntan lotions. Therefore, it is a potential low-toxicity antifungal agent that may be used topically because of its penetration into the corneal layers of the skin. PMID:20936361

  4. Synthesis of isatin thiosemicarbazones derivatives: In vitro anti-cancer, DNA binding and cleavage activities

    NASA Astrophysics Data System (ADS)

    Ali, Amna Qasem; Teoh, Siang Guan; Salhin, Abdussalam; Eltayeb, Naser Eltaher; Khadeer Ahamed, Mohamed B.; Majid, A. M. S. Abdul

    New derivatives of thiosemicarbazone Schiff base with isatin moiety were synthesized L1-L6. The structures of these compounds were characterized based on the spectroscopic techniques. Compound L6 was further characterized by XRD single crystal. The interaction of these compounds with calf thymus (CT-DNA) exhibited high intrinsic binding constant (kb = 5.03-33.00 × 105 M-1) for L1-L3 and L5 and (6.14-9.47 × 104 M-1) for L4 and L6 which reflect intercalative activity of these compounds toward CT-DNA. This result was also confirmed by the viscosity data. The electrophoresis studies reveal the higher cleavage activity of L1-L3 than L4-L6. The in vitro anti-proliferative activity of these compounds against human colon cancer cell line (HCT 116) revealed that the synthesized compounds (L3, L6 and L2) exhibited good anticancer potency.

  5. In vitro evaluation of synergistic activity between ciprofloxacin and broad snouted caiman serum against Escherichia coli.

    PubMed

    Siroski, P A; Russi, N B; Ortega, H H; Formentini, E A

    2015-02-01

    The in vitro synergistic activity between ciprofloxacin and serum of broad snouted caiman on Escherichia coli was studied. The estimated MIC value of ciprofloxacin was 0.0188?µg/ml, and two assays of kill curve during 5 hours were performed: the first one in a standard culture medium and the second one in the presence of caiman serum. Different concentrations of ciprofloxacin were tested. Ciprofloxacin showed higher values of bacterial elimination rate in the presence of caiman serum in all concentrations tested. The combined activity of sub-inhibitory concentrations of ciprofloxacin and the humoral immune factors present in caiman serum determined an increase in the bacterial elimination observed in this assay. We suggest that the antibacterial activity of complement and natural antibodies present in caiman serum, which can bind to both Gram-negative and Gram-positive bacteria and acting through the classical complement pathway, can inhibit bacterial growth of Escherichia coli by lysis. PMID:25468795

  6. Phytochemical constituents and in vitro radical scavenging activity of different Aloe species.

    PubMed

    Lucini, Luigi; Pellizzoni, Marco; Pellegrino, Roberto; Molinari, Gian Pietro; Colla, Giuseppe

    2015-03-01

    The phytochemical profile of Aloe barbadensis Mill. and Aloe arborescens Mill. was investigated using colorimetric assays, triple quadrupole and time-of-flight mass spectrometry, focusing on phenolic secondary metabolites in the different leaf portions. Hydroxycinnamic acids, several characteristic anthrones and chromones, the phenolic dimer feralolide and flavonoids such as flavones and isoflavones were identified. The stable radical DPPH test and the ORAC assay were then used to determine the in vitro radical scavenging. The outer green rind was the most active, while the inner parenchyma was much less effective. The 5-methylchromones aloesin, aloeresin A and aloesone were the most active among the pure secondary metabolites tested. The results suggest that several compounds are likely to contribute to the overall radical scavenging activity, and indicate that leaf portion must be taken into account when the plant is used for its antioxidant properties. PMID:25306376

  7. Synthesis of ribozyme against vascular endothelial growth factor165 and its biological activity in vitro

    PubMed Central

    Gu, Zhong-Ping; Wang, Yun-Jie; Wu, Yu; Li, Jin-Ge; Chen, Nong-An

    2004-01-01

    AIM: To investigate the designation, synthesis and biological activity of against vascular endothelial growth factor165 (VEGF165) ribozyme. METHODS: The ribozyme against VEGF165 was designed with computer. The transcriptional vector was constructed which included the anti-VEGF165 ribozyme and 5’, 3’ self-splicing ribozymes. The hammerhead ribozyme and substrate VEGF165 mRNA were synthesized through transcription in vitro. The cleavage activity of the ribozyme on target RNA was observed in a cell-free system. RESULTS: The anti-VEGF165 ribozyme was released properly from the transcription of pGEMRz212 cleaved by 5’ and 3’ self-splicing ribozymes which retained its catalytic activity, and the cleavage efficiency of ribozyme reached 90.7%. CONCLUSION: The anti-VEGF165 ribozyme designed with computer can cleave VEGF165 mRNA effectively. PMID:15133860

  8. In vitro inhibition activity of essential oils from some Lamiaceae species against phytopathogenic fungi.

    PubMed

    Kumar, Vinod; Mathela, C S; Tewari, A K; Bisht, K S

    2014-09-01

    Natural products have been in focus as alternative, effective and safe materials against the phytopathogens. Investigations show Nepeta oils as effective in controlling the food crops decay. The inhibitory effects of essential oils derived from Nepeta leucophylla, Nepeta ciliaris, Nepeta clarkei and Calamintha umbrosa against five phytopthogenic fungi have been determined. In vitro antifungal activity varied with their constituents and target species. More active being the oils containing oxygenated terpenoids. Helminthosporium maydis was sensitive to the all oils, IC50 values have 43.6-109.3 ?g mL(-1). The N. leucophylla oil possessing oxygenated iridoids was more effective against H. maydis (IC50 value of 43.6 ?g mL(-1)) while N. ciliaris was more active against Fusarium oxysporum (IC50 value of 219.2 ?g mL(-1)). The oils were effective against the spore germination of all the tested plant pathogens. PMID:25175652

  9. [In vitro activity of a liposomal nystatin formulation (Nyotran) against Cryptococcus neoformans].

    PubMed

    Alonso-Vargas, R; González-Alvarez, L; Ruesga, M T; Carrillo-Muńoz, A J; Martín-Mazuelos, E; Wallace, T L; Cossum, P A; Pontón, J; Quindós, G

    2000-09-01

    The in vitro antifungal activity of a new liposomal nystatin formulation (NISTL, Nyotran, Aronex Ltd., EE.UU.) was evaluated by a microdilution method with RPMI based on the M27A document of the National Committee for Clinical Laboratory Standards (NCCLS) against 22 isolates of Cryptococcus neoformans. This antifungal activity was compared with those of other seven antifungal agents, such as nystatin (NIST), amphotericin B deoxycholate, liposomal amphotericin B, amphotericin B lipid complex, amphotericin B colloidal dispersion, fluconazole, and itraconazole. NISTL was more active in vitrothan NIST, showing MIC values 2-3 fold smaller in 90% of the isolates. The results obtained suggest that this new formulation would be very helpful for the treatment of cryptococcosis. PMID:15762799

  10. New formula herbal pellets demonstrate a uniform and stable release of the active ingredients in vitro.

    PubMed

    Pranskuniene, Zivile; Bernatoniene, Jurga; Kalveniene, Zenona; Masteikova, Ruta; Mekas, Tauras; Velziene, Saule; Ivanauskas, Kostas; Suchockas, Vytautas; Mintauckiene, Inesa; Savickas, Arunas

    2013-01-01

    The aim of this study was to evaluate the effect of different capsule filling manufacturing techniques and storage conditions on the release of the active ingredients from herbal capsules during the dissolution test in vitro. Different techniques for the preparation of the original mixture of dry extracts were applied, and subsequently capsules with six different fillings were prepared. The stability of the capsules was evaluated in different long-term storage conditions, registering changes in the water content (loss of drying), capsule disintegration time, and phenolic compounds dissolution test in vitro. The baseline of phenolic compounds release in the control capsules (filled with the mixture of the powder of dry herbal extracts) was the highest, compared to other capsule groups, yet during long-term storage, these capsules accumulated too much moisture, which impeded capsule disintegration time and phenolic compounds release. The study showed that moisture and temperature changes occurring during the storage of the preparation had a negative effect on the release of phenolic compounds from herbal capsules. Capsules filled with pellets demonstrated a uniform and stable release of the active ingredients in different long-term storage conditions, which indicates that the manufacturing technology of dry herbal extracts affects the stability of the active ingredients. PMID:23923396

  11. Antitumor activity of Portulaca oleracea L. polysaccharides against cervical carcinoma in vitro and in vivo.

    PubMed

    Zhao, Rui; Gao, Xu; Cai, Yaping; Shao, Xingyue; Jia, Guiyan; Huang, Yulan; Qin, Xuegong; Wang, Jingwei; Zheng, Xiaoliang

    2013-07-25

    Portulaca oleracea L. has been used as folk medicine in different countries to treat different ailments in humans. P. oleracea L. polysaccharide (POL-P), extracted from P. oleracea L., is found to have bioactivities such as hypoglycemic and hypolipidemic activities, antioxidant and antitumor activities. In our study, a water-soluble polysaccharide (POL-P3b) was successfully purified from Galium verum L. by DEAE cellulose and Sephadex G-200 column chromatography. To evaluate the anticancer efficacy and associated mechanisms of POL-P3b on cervical cancer in vitro and in vivo, we showed that treatment of HeLa cell with POL-P3b inhibited cell proliferation. In addition, POL-P3b significantly inhibited tumor growth in U14-bearing mice. Further analysis indicated that POL-P3b possesses the activity of inhibiting cervical cancer cell growth in vitro and in vivo at a concentration- and time-dependent manner, and the mechanisms were associated with Sub-G1 phase cell cycle arrest, triggering DNA damage and inducing apoptosis. PMID:23768576

  12. In vitro antioxidant and antidiabetic activities of biomodified lignin from Acacia nilotica wood.

    PubMed

    Barapatre, Anand; Aadil, Keshaw Ram; Tiwary, Bhupendra Nath; Jha, Harit

    2015-04-01

    The antioxidant and antidiabetic activity of biomodified alkali lignin extracted from a deciduous plant Acacia nilotica, was evaluated in vitro. The extracted alkali lignin was subjected to microbial biotransformation by ligninolytic fungus Aspergillus flavus and Emericella nidulans. These modifications were done under varying concentration of carbon to nitrogen sources. The structural feature of the lignin samples were compared by FTIR, functional group analysis and (13)C solid state NMR. All lignin samples were tested for antioxidant efficiency, reducing power and H2O2 scavenging power. Modifications in all lignin samples showed correlation with their antioxidant scavenging activity and reducing power. Antidiabetic properties were evaluated in terms of in vitro glucose movement inhibition and ?-amylase inhibition assay. Modified samples exhibited increased glucose binding efficiency as demonstrated by the decreased glucose diffusion (55.5-76.3%) and 1.16-1.18-fold enhanced ?-amylase inhibition in comparison to their control samples. The results obtained demonstrate that the structure and functional modifications in lignin significantly affects its bioefficacy in term of antioxidant and antidiabetic activities. PMID:25600985

  13. In vitro cytotoxic activity of chitosan-bullfrog oil microemulsion against melanoma cells.

    PubMed

    Bonatto, Cínthia Caetano; Joanitti, Graziella Anselmo; Silva, Luciano Paulino

    2015-08-01

    Microemulsion-based animal oils, alone or associated with polymers have been extensively used in pharmacy, medicine and cosmetics, since the major lipid constituents of the oils show several biological activities. Despite showing antimicrobial activity, there are no reports in the literature regarding the effects of bullfrog oil on cytotoxic activity against tumor cells. The aim of the present study was to synthesize, characterise and evaluate the in vitro effects on melanoma cell line (B16F10) of bullfrog oil microemulsions associated or not with chitosan, surfactant and bullfrog oil (CSBO) and surfactant and bullfrog oil (SBO), respectively. The microemulsions were developed and their physical-chemical characteristics were evaluated by light microscopy, dynamic light scattering, atomic force microscopy and zeta potential. The microemulsions showed regular spherical shapes, high polydispersity and excellent (+82.2 ± 1.0 mV) to low (-16.0 ± 0.5 mV), colloidal stability. The systems significantly decreased the in vitro cell viability of melanoma skin cancer by up to 90.2% (CSBO) and 91.8% (SBO); while free bullfrog oil showed no effects. The results obtained from microemulsions of bullfrog oil indicate the potential of the microemulsions developed, alone or in combination with other chemotherapeutic agents, for future use in biomedical approaches aiming towards cancer therapy. PMID:26224345

  14. Bluetongue Virus VP1 Polymerase Activity In Vitro: Template Dependency, Dinucleotide Priming and Cap Dependency

    PubMed Central

    Matsuo, Eiko; Roy, Polly

    2011-01-01

    Background Bluetongue virus (BTV) protein, VP1, is known to possess an intrinsic polymerase function, unlike rotavirus VP1, which requires the capsid protein VP2 for its catalytic activity. However, compared with the polymerases of other members of the Reoviridae family, BTV VP1 has not been characterized in detail. Methods and Findings Using an in vitro polymerase assay system, we demonstrated that BTV VP1 could synthesize the ten dsRNAs simultaneously from BTV core-derived ssRNA templates in a single in vitro reaction as well as genomic dsRNA segments from rotavirus core-derived ssRNA templates that possess no sequence similarity with BTV. In contrast, dsRNAs were not synthesized from non-viral ssRNA templates by VP1, unless they were fused with specific BTV sequences. Further, we showed that synthesis of dsRNAs from capped ssRNA templates was significantly higher than that from uncapped ssRNA templates and the addition of dinucleotides enhanced activity as long as the last base of the dinucleotide complemented the 3? -terminal nucleotide of the ssRNA template. Conclusions We showed that the polymerase activity was stimulated by two different factors: cap structure, likely due to allosteric effect, and dinucleotides due to priming. Our results also suggested the possible presence of cis-acting elements shared by ssRNAs in the members of family Reoviridae. PMID:22110731

  15. In vitro and in vivo antioxidant activity of a water-soluble polysaccharide from dendrobium denneanum

    USGS Publications Warehouse

    Luo, A.; Ge, Z.; Fan, Y.; Chun, Z.; Jin, He X.

    2011-01-01

    The water-soluble crude polysaccharide (DDP) obtained from the aqueous extracts of the stem of Dendrobium denneanum through hot water extraction followed by ethanol precipitation, was found to have an average molecular weight (Mw) of about 484.7 kDa. Monosaccharide analysis revealed that DDP was composed of arabinose, xylose, mannose, glucose and galactose in a molar ratio of 1.00:2.66:8.92:34.20:10.16. The investigation of antioxidant activity both in vitro and in vivo showed that DDP is a potential antioxidant. ?? 2011.

  16. Polyoxygenated Steroids from the Octocoral Leptogorgia punicea and in Vitro Evaluation of Their Cytotoxic Activity

    PubMed Central

    Moritz, Maria Izabel G.; Marostica, Lucas Lourenço; Bianco, Éverson M.; Almeida, Maria Tereza R.; Carraro, Joăo L.; Cabrera, Gabriela M.; Palermo, Jorge A.; Simőes, Cláudia M. O.; Schenkel, Eloir P.

    2014-01-01

    Five new polyoxygenated marine steroids—punicinols A–E (1–5)—were isolated from the gorgonian Leptogorgia punicea and characterized by spectroscopic methods (IR, MS, 1H, 13C and 2-D NMR). The five compounds induced in vitro cytotoxic effects against lung cancer A549 cells, while punicinols A and B were the most active, with IC50 values of 9.7 ?M and 9.6 ?M, respectively. The synergistic effects of these compounds with paclitaxel, as well as their effects on cell cycle distribution and their performance in the clonogenic assay, were also evaluated. Both compounds demonstrated significant synergistic effects with paclitaxel. PMID:25486111

  17. Flavonol dimers from callus cultures of Dysosma versipellis and their in vitro neuraminidase inhibitory activities.

    PubMed

    Chen, Ridao; Duan, Ruigang; Wei, Yannan; Zou, Jianhua; Li, Junwei; Liu, Xiaoyue; Wang, Haiyan; Guo, Ying; Li, Qiuhong; Dai, Jungui

    2015-12-01

    A chemical investigation of callus cultures of Dysosma versipellis led to the isolation of five new flavonol dimers, dysoverines A-E (1-5), together with 12 known compounds (6-17). The structures of new compounds were determined by the extensive spectroscopic data analyses. The biosynthetic pathway of the new compounds was proposed to involve O-methylation, prenylation, and Diels-Alder cycloaddition, which successively occurred in cultured plant cells. Compounds 1-17 exhibited in vitro neuraminidase inhibitory activities with the IC50 values of 31.0-93.9?M. PMID:26481138

  18. Comparative in vitro activity profile of oritavancin against recent gram-positive clinical isolates.

    PubMed

    Arhin, Francis F; Draghi, Deborah C; Pillar, Chris M; Parr, Thomas R; Moeck, Gregory; Sahm, Daniel F

    2009-11-01

    Oritavancin activity was tested against 15,764 gram-positive isolates collected from 246 hospital centers in 25 countries between 2005 and 2008. Organisms were Staphylococcus aureus (n = 9,075), coagulase-negative staphylococci (n = 1,664), Enterococcus faecalis (n = 1,738), Enterococcus faecium (n = 819), Streptococcus pyogenes (n = 959), Streptococcus agalactiae (n = 415), group C, G, and F streptococci (n = 84), and Streptococcus pneumoniae (n = 1,010). Among the evaluated staphylococci, 56.7% were resistant to oxacillin. The vancomycin resistance rate among enterococci was 21.2%. Penicillin-resistant and -intermediate rates were 14.7% and 21.4%, respectively, among S. pneumoniae isolates. Among nonpneumococcal streptococci, 18.5% were nonsusceptible to erythromycin. Oritavancin showed substantial in vitro activity against all organisms tested, regardless of resistance profile. The maximum oritavancin MIC against all staphylococci tested (n = 10,739) was 4 microg/ml; the MIC(90) against S. aureus was 0.12 microg/ml. Against E. faecalis and E. faecium, oritavancin MIC(90)s were 0.06 and 0.12, respectively. Oritavancin was active against glycopeptide-resistant enterococci, including VanA strains (n = 486), with MIC(90)s of 0.25 and 1 microg/ml against VanA E. faecium and E. faecalis, respectively. Oritavancin showed potent activity against streptococci (n = 2,468); MIC(90)s for the different streptococcal species were between 0.008 and 1 microg/ml. These data are consistent with previous studies with respect to resistance rates of gram-positive isolates and demonstrate the spectrum and in vitro activity of oritavancin against a wide variety of contemporary gram-positive pathogens, regardless of resistance to currently used drugs. The data provide a foundation for interpreting oritavancin activity and potential changes in susceptibility over time once oritavancin enters into clinical use. PMID:19738026

  19. Comparative In Vitro Activity Profile of Oritavancin against Recent Gram-Positive Clinical Isolates?

    PubMed Central

    Arhin, Francis F.; Draghi, Deborah C.; Pillar, Chris M.; Parr, Thomas R.; Moeck, Gregory; Sahm, Daniel F.

    2009-01-01

    Oritavancin activity was tested against 15,764 gram-positive isolates collected from 246 hospital centers in 25 countries between 2005 and 2008. Organisms were Staphylococcus aureus (n = 9,075), coagulase-negative staphylococci (n = 1,664), Enterococcus faecalis (n = 1,738), Enterococcus faecium (n = 819), Streptococcus pyogenes (n = 959), Streptococcus agalactiae (n = 415), group C, G, and F streptococci (n = 84), and Streptococcus pneumoniae (n = 1,010). Among the evaluated staphylococci, 56.7% were resistant to oxacillin. The vancomycin resistance rate among enterococci was 21.2%. Penicillin-resistant and -intermediate rates were 14.7% and 21.4%, respectively, among S. pneumoniae isolates. Among nonpneumococcal streptococci, 18.5% were nonsusceptible to erythromycin. Oritavancin showed substantial in vitro activity against all organisms tested, regardless of resistance profile. The maximum oritavancin MIC against all staphylococci tested (n = 10,739) was 4 ?g/ml; the MIC90 against S. aureus was 0.12 ?g/ml. Against E. faecalis and E. faecium, oritavancin MIC90s were 0.06 and 0.12, respectively. Oritavancin was active against glycopeptide-resistant enterococci, including VanA strains (n = 486), with MIC90s of 0.25 and 1 ?g/ml against VanA E. faecium and E. faecalis, respectively. Oritavancin showed potent activity against streptococci (n = 2,468); MIC90s for the different streptococcal species were between 0.008 and 1 ?g/ml. These data are consistent with previous studies with respect to resistance rates of gram-positive isolates and demonstrate the spectrum and in vitro activity of oritavancin against a wide variety of contemporary gram-positive pathogens, regardless of resistance to currently used drugs. The data provide a foundation for interpreting oritavancin activity and potential changes in susceptibility over time once oritavancin enters into clinical use. PMID:19738026

  20. In vitro and in vivo activities of piritetrate (M-732), a new antidermatophytic thiocarbamate.

    PubMed Central

    Iwata, K; Yamashita, T; Uehara, H

    1989-01-01

    Piritetrate (M-732), a new topical antifungal agent belonging chemically to the thiocarbamates, was demonstrated to possess a potent selective antidermatophytic activity. In terms of its MICs in susceptibility testing, mainly done by using Sabouraud dextrose agar plates, piritetrate exhibited several- to 10-fold-stronger antidermatophytic activity than tolnaftate, a reference thiocarbamate. Furthermore, piritetrate was found to show a broader antifungal spectrum than tolnaftate; relatively many species and strains of dematiaceous fungi, dimorphic fungi, and some other filamentous fungi as well as a few strains of Cryptococcus neoformans were fairly susceptible to piritetrate, while almost all the tested species and strains were resistant to tolnaftate. All the tested species of the genus Candida were, however, resistant to both compounds. Variables which can influence antimicrobial activity caused few changes in the MICs of either compound against Trichophyton mentagrophytes; however, an increase in the inoculum size resulted in a significant increase in the MICs. The antidermatophytic activities of piritetrate and tolnaftate were fungistatic but not fungicidal. Piritetrate also exhibited a more potent in vitro anti-T. mentagrophytes activity than clotrimazole or tolciclate. Piritetrate and tolnaftate had no antibacterial activity. The in vivo activity of topically administered piritetrate against experimental dermal infection of guinea pigs with T. mentagrophytes was more effective than that of tolnaftate both mycologically and clinically. Piritetrate manifested no acute toxicity in laboratory animals when administered even in large quantities by the oral, intraperitoneal, and topical routes. PMID:2619275

  1. Instrument and technique for the in vitro screening of platelet activation from whole blood samples

    NASA Astrophysics Data System (ADS)

    Martin, Yves; Lépine, Mariette; Bannari, Abdelfettah; Vermette, Patrick

    2007-05-01

    The measurement of platelet activation is very difficult to accomplish clinically as platelets are readily activated by in vitro manipulations. Although techniques such as platelet aggregation and flow cytometry exist to estimate platelet function, important limitations prevent these techniques to be widely accepted. In this study, low-fouling surfaces used to limit ex vivo platelet activation were locally bioactivated to rapidly detect platelet activation from whole blood through the selective local adhesion and aggregation of artificially activated platelets. To achieve this result, a fabrication method was developed to create arrays of anti-CD62 and anti-CD61 proteins covalently immobilized on substrates covered by low-fouling graft layers. Moreover, to further limit ex vivo platelet activation and to obtain reproducible results, a custom-made flow chamber was designed and fabricated with the help of computer-assisted mathematical modeling to create defined shear environments. This diagnostic instrument has the potential to allow the rapid estimation of platelet activation levels in whole blood.

  2. In vitro activity of E1210, a novel antifungal, against clinically important yeasts and molds.

    PubMed

    Miyazaki, Mamiko; Horii, Takaaki; Hata, Katsura; Watanabe, Nao-Aki; Nakamoto, Kazutaka; Tanaka, Keigo; Shirotori, Syuji; Murai, Norio; Inoue, Satoshi; Matsukura, Masayuki; Abe, Shinya; Yoshimatsu, Kentaro; Asada, Makoto

    2011-10-01

    E1210 is a new antifungal compound with a novel mechanism of action and broad spectrum of antifungal activity. We investigated the in vitro antifungal activities of E1210 compared to those of fluconazole, itraconazole, voriconazole, amphotericin B, and micafungin against clinical fungal isolates. E1210 showed potent activities against most Candida spp. (MIC(90) of ?0.008 to 0.06 ?g/ml), except for Candida krusei (MICs of 2 to >32 ?g/ml). E1210 showed equally potent activities against fluconazole-resistant and fluconazole-susceptible Candida strains. E1210 also had potent activities against various filamentous fungi, including Aspergillus fumigatus (MIC(90) of 0.13 ?g/ml). E1210 was also active against Fusarium solani and some black molds. Of note, E1210 showed the greatest activities against Pseudallescheria boydii (MICs of 0.03 to 0.13 ?g/ml), Scedosporium prolificans (MIC of 0.03 ?g/ml), and Paecilomyces lilacinus (MICs of 0.06 ?g/ml) among the compounds tested. The antifungal action of E1210 was fungistatic, but E1210 showed no trailing growth of Candida albicans, which has often been observed with fluconazole. In a cytotoxicity assay using human HK-2 cells, E1210 showed toxicity as low as that of fluconazole. Based on these results, E1210 is likely to be a promising antifungal agent for the treatment of invasive fungal infections. PMID:21825291

  3. In Vitro Activity of Rifampicin Combined with Daptomycin or Tigecycline on Staphylococcus haemolyticus Biofilms.

    PubMed

    Szczuka, Ewa; Grabska, Katarzyna; Kaznowski, Adam

    2015-08-01

    Staphylococcus haemolyticus is of increasing concern as a cause of several biofilm-associated infections, and today, it represents the second most common organism among clinical isolates of coagulase-negative staphylococci. However, little is known regarding the treatment of infections caused by these bacteria. In this study, we characterize the biofilm formed by S. haemolyticus strains isolated from bloodstream infections and assess in vitro the activity of rifampicin combined with daptomycin or tigecycline against bacteria growing in a biofilm. The results of our studies indicated that the majority (78 %) of methicillin-resistant Staphylococcus haemolyticus strains have the ability to form a biofilm in vitro. None of these strains carried icaADBC genes indicating that they form biofilm via ica-independent mechanisms. The molecular characterization of the biofilm showed that proteins are the predominant matrix component and play a major role in biofilm structure. Extracellular DNA and polysaccharides, other than polysaccharide intercellular adhesin, are also present in the biofilm matrix, but they play a minor role. The images obtained by confocal laser scanning microscopy showed that most S. haemolyticus strains formed a dense biofilm with a low number of dead cells. In vitro study demonstrated excellent activity of tigecycline in combination with rifampicin against cell growth in the proteinous biofilm. The BIC (biofilm inhibitory concentration) value for tigecycline/rifampicin ranged from 0.062 to 1 µg/ml, whereas for daptomycin/rifampicin from 0.125 to 2 µg/ml. These results indicated that the tigecycline/rifampicin combination was more effective against ica-independent biofilm, formed by S. haemolyticus strains, than the daptomycin/rifampicin combination. PMID:25894996

  4. In vitro activity of origanum vulgare essential oil against candida species

    PubMed Central

    Cleff, Marlete Brum; Meinerz, Ana Raquel; Xavier, Melissa; Schuch, Luiz Filipe; Schuch, Luiz Filipe; Araújo Meireles, Mário Carlos; Alves Rodrigues, Maria Regina; de Mello, Joăo Roberto Braga

    2010-01-01

    The aim of this study was to evaluate the in vitro activity of the essential oil extracted from Origanum vulgare against sixteen Candida species isolates. Standard strains tested comprised C. albicans (ATCC strains 44858, 4053, 18804 and 3691), C. parapsilosis (ATCC 22019), C. krusei (ATCC 34135), C. lusitaniae (ATCC 34449) and C. dubliniensis (ATCC MY646). Six Candida albicans isolates from the vaginal mucous membrane of female dogs, one isolate from the cutaneous tegument of a dog and one isolate of a capuchin monkey were tested in parallel. A broth microdilution technique (CLSI) was used, and the inoculum concentration was adjusted to 5 x 106 CFU mL-1. The essential oil was obtained by hydrodistillation in a Clevenger apparatus and analyzed by gas chromatography. Susceptibility was expressed as Minimal Inhibitory Concentration (MIC) and Minimal Fungicidal Concentration (MFC). All isolates tested in vitro were sensitive to O. vulgare essential oil. The chromatographic analysis revealed that the main compounds present in the essential oil were 4-terpineol (47.95%), carvacrol (9.42%), thymol (8.42%) and ?-terpineol (7.57%). C. albicans isolates obtained from animal mucous membranes exhibited MIC and MFC values of 2.72 ?L mL-1 and 5 ?L mL-1, respectively. MIC and MFC values for C. albicans standard strains were 2.97 ?L mL-1 and 3.54 ?L mL-1, respectively. The MIC and MFC for non-albicans species were 2.10 ?L mL-1 and 2.97 ?L mL-1, respectively. The antifungal activity of O. vulgare essential oil against Candida spp. observed in vitro suggests its administration may represent an alternative treatment for candidiasis. PMID:24031471

  5. Estrous sheep serum enables in vitro capacitation of ram spermatozoa while preventing caspase activation.

    PubMed

    Del Olmo, E; García-Álvarez, O; Maroto-Morales, A; Ramón, M; Jiménez-Rabadán, P; Iniesta-Cuerda, M; Anel-Lopez, L; Martinez-Pastor, F; Soler, A J; Garde, J J; Fernández-Santos, M R

    2016-01-15

    Estrous sheep serum (ESS) is considered the most efficient agent for in vitro capacitation of ram spermatozoa. We have explored the relationship between caspase activation and capacitation in ram. Semen samples from 17 rams were cryopreserved. In vivo fertility was evaluated after intrauterine artificial insemination. Samples were submitted to four treatments: control, ESS (10%), caspase inhibitor (Z-VAD-FMK), and estrous ewe serum plus caspase inhibitor (I + E). Sperm samples were incubated for 30 minutes at 38.5 °C and 5% CO2 and analyzed with flow cytometry for mitochondrial membrane potential (MitoTracker deep red), sperm viability and apoptosis-like changes (YO-PRO-1/propidium iodide), acrosomal status (peanut agglutinin-fluorescein isothiocyanate), membrane fluidity (merocyanine 540), and caspase activity (Vybrant FAM kits for polycaspases, caspase-8, and caspases 3-7). Estrous sheep serum induced changes compatible with capacitation, doubling the proportion of viable spermatozoa with increased merocyanine 540 and increasing YO-PRO-1(+) and acrosome-reacted spermatozoa (P < 0.05). Incubation increased the proportion of spermatozoa with activated caspases (P < 0.05), which was abolished by the treatments. We detected a simultaneous decrease in the proportion of the viable and caspase(-) spermatozoa after the incubation, which was prevented by the presence of estrous ewe serum (P < 0.05). The analysis of caspases 3/7 and 8 resulted in less marked differences. Fertility was positively related to viability and inactivated caspases and negatively to viable-capacitated spermatozoa and active caspases. In vitro induction of capacitation in thawed ram spermatozoa by using ESS suggests a downregulation in apoptotic pathways. However, males with the lowest fertility showed parameters similar to high-fertility males, suggesting that other factors were involved apart from capacitation and/or caspase activation. PMID:26474680

  6. In vitro activity of almond skin polyphenols for scavenging free radicals and inducing quinone reductase.

    PubMed

    Chen, C-Y Oliver; Blumberg, Jeffrey B

    2008-06-25

    Observational studies and clinical trials suggest nut intake, including almonds, is associated with an enhancement in antioxidant defense and a reduction in the risk of cancer and cardiovascular disease. Almond skins are rich in polyphenols (ASP) that may contribute to these putative benefits. To assess their potential mechanisms of action, we tested the in vitro effect of ASP extracted with methanol (M) or a gastrointestinal juice mimic (GI) alone or in combination with vitamins C (VC) or E (VE) (1-10 micromol/L) on scavenging free radicals and inducing quinone reductase (QR). Flavonoid profiles from ASP-M and -GI extracts were different from one another. ASP-GI was more potent in scavenging HOCl and ONOO (-) radicals than ASP-M. In contrast, ASP-M increased and ASP-GI decreased QR activity in Hepa1c1c7 cells. Adding VC or VE to ASP produced a combination- and dose-dependent action on radical scavenging and QR induction. In comparison to their independent actions, ASP-M plus VC were less potent in scavenging DPPH, HOCl, ONOO (-), and O 2 (-) (*). However, the interaction between ASP-GI plus VC promoted their radical scavenging activity. Combining ASP-M plus VC resulted in a synergistic interaction, inducing QR activity, but ASP-GI plus VC had an antagonistic effect. On the basis of their total phenolic content, the measures of total antioxidant activity of ASP-M and -GI were comparable. Thus, in vitro, ASP act as antioxidants and induce QR activity, but these actions are dependent upon their dose, method of extraction, and interaction with antioxidant vitamins. PMID:18512942

  7. Leptin activates chicken growth hormone promoter without chicken STAT3 in vitro.

    PubMed

    Murase, Daisuke; Namekawa, Shoko; Ohkubo, Takeshi

    2016-01-01

    Leptin is an adipocyte-derived hormone that not only regulates food intake and energy homeostasis but also induces growth hormone (GH) mRNA expression and release, thereby controlling growth and metabolism in mammals. The molecular mechanism of leptin-induced regulation of GH gene transcription is unclear. The current study investigated the effects of leptin on the chicken GH (cGH) promoter and the molecular mechanism underlying leptin-induced cGH gene expression in vitro. Leptin activated the cGH promoter in the presence of chPit-1? in CHO cells stably expressing the chicken leptin receptor. Promoter activation did not require STAT-binding elements in the cGH promoter or STAT3 activity. However, JAK2 activation was required for leptin-dependent activity. JAK2-dependent pathways include p42/44 MAPK and PI3K, and inhibition of these pathways partially blocked leptin-induced cGH gene transcription. Although CK2 directly activates JAK2, a CK2 inhibitor blocked leptin-dependent activation of the cGH gene without affecting JAK2 phosphorylation. The CK2 inhibitor suppressed Erk1/2 and Akt phosphorylation. Additional data implicate Src family kinases in leptin-dependent cGH gene activation. These results suggest that leptin activates the cGH gene in the presence of chPit-1? via several leptin-activated kinases. Although further study is required, we suggest that the leptin-induced JAK2/p42/44 MAPK and JAK2/PI3K cascades are activated by Src-meditated CK2, leading to CBP phosphorylation and interaction with chPit-1?, resulting in transactivation of the cGH promoter. PMID:26403688

  8. Uridine 5?-Triphosphate Promotes In Vitro Schwannoma Cell Migration through Matrix Metalloproteinase-2 Activation

    PubMed Central

    Martiańez, Tania; Segura, Mňnica; Figueiro-Silva, Joana; Grijota-Martinez, Carmen; Trullas, Ramón; Casals, Núria

    2014-01-01

    In response to peripheral nerve injury, Schwann cells adopt a migratory phenotype and modify the extracellular matrix to make it permissive for cell migration and axonal re-growth. Uridine 5?-triphosphate (UTP) and other nucleotides are released during nerve injury and activate purinergic receptors expressed on the Schwann cell surface, but little is known about the involvement of purine signalling in wound healing. We studied the effect of UTP on Schwannoma cell migration and wound closure and the intracellular signaling pathways involved. We found that UTP treatment induced Schwannoma cell migration through activation of P2Y2 receptors and through the increase of extracellular matrix metalloproteinase-2 (MMP-2) activation and expression. Knockdown P2Y2 receptor or MMP-2 expression greatly reduced wound closure and MMP-2 activation induced by UTP. MMP-2 activation evoked by injury or UTP was also mediated by phosphorylation of all 3 major mitogen-activated protein kinases (MAPKs): JNK, ERK1/2, and p38. Inhibition of these MAPK pathways decreased both MMP-2 activation and cell migration. Interestingly, MAPK phosphorylation evoked by UTP exhibited a biphasic pattern, with an early transient phosphorylation 5 min after treatment, and a late and sustained phosphorylation that appeared at 6 h and lasted up to 24 h. Inhibition of MMP-2 activity selectively blocked the late, but not the transient, phase of MAPK activation. These results suggest that MMP-2 activation and late MAPK phosphorylation are part of a positive feedback mechanism to maintain the migratory phenotype for wound healing. In conclusion, our findings show that treatment with UTP stimulates in vitro Schwannoma cell migration and wound repair through a MMP-2-dependent mechanism via P2Y2 receptors and MAPK pathway activation. PMID:24905332

  9. Ethnopharmacological in vitro studies on Austria's folk medicine—An unexplored lore in vitro anti-inflammatory activities of 71 Austrian traditional herbal drugs?

    PubMed Central

    Vogl, Sylvia; Picker, Paolo; Mihaly-Bison, Judit; Fakhrudin, Nanang; Atanasov, Atanas G.; Heiss, Elke H.; Wawrosch, Christoph; Reznicek, Gottfried; Dirsch, Verena M.; Saukel, Johannes; Kopp, Brigitte

    2013-01-01

    Ethnopharmacological relevance In Austria, like in most Western countries, knowledge about traditional medicinal plants is becoming scarce. Searching the literature concerning Austria's ethnomedicine reveals its scant scientific exploration. Aiming to substantiate the potential of medicinal plants traditionally used in Austria, 63 plant species or genera with claimed anti-inflammatory properties listed in the VOLKSMED database were assessed for their in vitro anti-inflammatory activity. Material and methods 71 herbal drugs from 63 plant species or genera were extracted using solvents of varying polarities and subsequently depleted from the bulk constituents, chlorophylls and tannins to avoid possible interferences with the assays. The obtained 257 extracts were assessed for their in vitro anti-inflammatory activity. The expression of the inflammatory mediators E-selectin and interleukin-8 (IL-8), induced by the inflammatory stimuli tumor necrosis factor alpha (TNF-?) and the bacterial product lipopolysaccharide (LPS) was measured in endothelial cells. The potential of the extracts to activate the nuclear factors PPAR? and PPAR? and to inhibit TNF-?-induced activation of the nuclear factor-kappa B (NF-?B) in HEK293 cells was determined by luciferase reporter gene assays. Results In total, extracts from 67 of the 71 assessed herbal drugs revealed anti-inflammatory activity in the applied in vitro test systems. Thereby, 30 could downregulate E-selectin or IL-8 gene expression, 28 were strong activators of PPAR? or PPAR? (inducing activation of more than 2-fold at a concentration of 10 µg/mL) and 21 evoked a strong inhibition of NF-?B (inhibition of more than 80% at 10 µg/mL). Conclusion Our research supports the efficacy of herbal drugs reported in Austrian folk medicine used for ailments associated with inflammatory processes. Hence, an ethnopharmacological screening approach is a useful tool for the discovery of new drug leads. PMID:23770053

  10. In vitro synergistic activity of antibiotic combinations against Brucella melitensis using E-test methodology.

    PubMed

    Kilic, Selcuk; Dizbay, Murat; Hizel, Kenan; Arman, Dilek

    2008-04-01

    The treatment of brucellosis is still problematic, because of high rates of treatment failure or relapses. As the microorganism is an intracellular pathogen, treatment requires combined regimens. However, limited existing data on in vitro combinations are avaliable for Brucellae. The aim of this study was to investigate the in vitro efficacy of various traditional and new antibiotic combinations against 16 Brucella melitensis strains. The combination effect of antimicrobial agents was evaluated by E-test synergy method to obtain a fractional inhibitory concentration (FIC) index. Co-Trimoxazole (SXT) and moxifloxocin (MXF) exhibited the lowest MIC, while Rifampin (RIF) had the highest MIC in the study. Combinations with RIF showed the best synergistic activity (100% of RIF-tetracycline (TET), and 87.5% of RIF-SXT). Synergistic activity was also detected at seven (43.7%) of ciprofloxocin (CIP)-SXT, four (25%) of TET-MXF, and two (12.5%) of TET-SXT combinations. The combinations that demonstrated additivity were TET-SXT, CIP-SXT and TET-MXF. Antagonism was observed only with the TET-Streptomycin (STR) combination in three strains (18.8%). Further work including randomized controlled clinical trials is required to fully evaluate the usefulness of these data. PMID:24031207

  11. In Vitro Activity of Tea Tree Oil Vaginal Suppositories against Candida spp. and Probiotic Vaginal Microbiota.

    PubMed

    Di Vito, Maura; Mattarelli, Paola; Modesto, Monica; Girolamo, Antonietta; Ballardini, Milva; Tamburro, Annunziata; Meledandri, Marcello; Mondello, Francesca

    2015-10-01

    The aim of this work is to evaluate the in vitro microbicidal activity of vaginal suppositories (VS) containing tea tree oil (TTO-VS) towards Candida spp. and vaginal probiotics. A total of 20 Candida spp. strains, taken from patients with vaginitis and from an established type collection, including reference strains, were analysed by using the CLSI microdilution method. To study the action of VS towards the beneficial vaginal microbiota, the sensitivity of Bifidobacterium animalis subsp. lactis (DSM 10140) and Lactobacillus spp. (Lactobacillus casei R-215 and Lactobacillus acidophilus R-52) was tested. Both TTO-VS and TTO showed fungicidal activity against all strains of Candida spp. whereas placebo-VS or the Aloe gel used as controls were ineffective. The study of fractional fungicidal concentrations (FFC) showed synergistic interaction with the association between Amphotericin B and TTO (0.25 to 0.08?µg/ml, respectively) against Candida albicans. Instead, the probiotics were only affected by TTO concentration???4% v/v, while, at concentrations?vitro, a selective fungicidal action, slightly affecting only the Bifidobacteriun animalis strain growth belonging to the vaginal microbiota. In vivo studies are needed to confirm the efficacy to prevent acute or recurrent vaginal candidiasis. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26235937

  12. Antiangiogenic activity of low-temperature lysozyme from a marine bacterium in vivo and in vitro

    NASA Astrophysics Data System (ADS)

    Wang, Zhenhua; Liu, Jincheng; Su, Ai; Sun, Mi; Wang, Chunbo

    2009-11-01

    We extracted marine low-temperature lysozyme (MLTL), a novel lysozyme, from a marine microorganism through fermentation. Our previous study suggested that a low molecular weight (16 kDa) may exert anti-tumor activity through antiangiogenesis. In this study, we extracted a high weight (39 kDa) and investigated its antiangiogenic activity in vivo and in vitro. Using zebrafish embryos as an in vivo study model, we found that treatment with MLTL significantly inhibited the growth of subintestinal vessels (SIVs) in a dose-dependent manner and that 400 µg/ml MLTL was sufficient to block the growth of SIVs. An in vitro study conducted using human umbilical vein endothelial cells (HUVECs) revealed that MLTL suppressed the proliferation, migration and tube formation of HUVECs in a dose-dependent manner. Interestingly, assays by flow cytometry and DNA electrophoresis indicated that MLTL was able to induce apoptosis of HUVECs. Moreover, further study demonstrated that the disruption of intracellular Ca2+ homeostasis may play an important role in MLTL induced apoptosis of HUVECs. Taken together, the results of this study demonstrate for the first time that MLTL inhibits angiogenesis through its pleiotropic effects on vascular endothelial cells and induces apoptosis through regulation of cellular Ca2+ levels. The results of this study also revealed a possible mechanism underlying the antiangiogenic effect of MLTL and suggested that MLTL may be a promising new antiangiogenic agent for use in cancer therapy.

  13. The Immunological Enhancement Activity of Propolis Flavonoids Liposome In Vitro and In Vivo

    PubMed Central

    Tao, Yang; Wang, Deqing; Hu, Yuanliang; Huang, Yee; Yu, Yun; Wang, Deyun

    2014-01-01

    The aim of this study was to investigate and assess the effects of propolis flavonoids liposome imposed on the immune system by comparing it to propolis flavonoids and blank liposome. In vitro, the effects of the above drugs on macrophages were assessed by measuring the phagocytic function and cytokine production. In vivo, the immunological adjuvant activity of propolis flavonoids liposome was compared with those of propolis flavonoids and blank liposome. The results showed that in vitro propolis flavonoids liposome can significantly enhance the phagocytic function of macrophages and the release of IL-1?, IL-6, and IFN-?. In addition, subcutaneous administration of propolis flavonoids liposome with ovalbumin to mice could effectively activate the cellular and humoral immune response, including inducing higher level concentrations of IgG, IL-4, and IFN-? in serum and the proliferation rates of splenic lymphocytes. These findings provided valuable information regarding the immune modulatory function of propolis flavonoids liposome and indicated the possibility of use of propolis flavonoids liposome as a potential adjuvant. PMID:25383082

  14. Physapubescin B Exhibits Potent Activity against Human Prostate Cancer In Vitro and In Vivo.

    PubMed

    Ding, Wanjing; Hu, Zhijuan; Zhang, Zhewen; Ma, Qiaoqiao; Tang, Huifang; Ma, Zhongjun

    2015-11-01

    The present data showed that a natural compound isolated from the plant Physalis pubescens L. (Solanaceae), physapubescin B, exhibited antitumor activity against prostate cancer in vitro and in vivo. Treating prostate cancer cells with physapubescin B resulted in the accumulation of cells in the G2/M phase, which was associated with reduced Cdc25C levels and increased levels of CyclinB1, P21 as well as p-Cdk1 (Tyr15). Additionally, reactive oxygen species (ROS) generation was increased in physapubescin B-treated PC-3 cells. Furthermore, the physapubescin B-induced decrease of Cdc25C protein expression together with the G2/M phase cell cycle arrest were significantly abrogated by antioxidant NAC and GSH. Our data also demonstrated that physapubescin B exhibited strong in vivo antitumor efficacy in human prostate cancer PC3 xenograft. In conclusion, our results provide clear evidence that physapubescin B exhibits antitumor activity both in vitro and in vivo and deserves further development as an anticancer agent. PMID:26415552

  15. In Vitro and in Vivo Studies of the Metabolic Activation of 8-Epidiosbulbin E Acetate.

    PubMed

    Lin, Dongju; Guo, Xiucai; Gao, Huiyuan; Cheng, Li; Cheng, Maosheng; Song, Shaojiang; Peng, Ying; Zheng, Jiang

    2015-09-21

    Furanoid 8-epidiosbulbin E acetate (EEA) is a major constituent of herbal medicine Dioscorea bulbifera L. (DB), a traditional Chinese medicine herb. Our preliminary studies demonstrated that administration of EEA caused acute hepatotoxicity in mice, and the observed toxicity required cytochromes P450-mediated metabolism. Metabolic activation studies of EEA were performed in vitro and in vivo. Microsomal incubations of EEA supplemented with N-acetyl lysine (NAL) and glutathione (GSH) generated six metabolites (M1-M6). M1-M4 were characterized as pyrrole derivatives, and M5 and M6 were pyrrolinones. M2-M6 were detected in bile and/or urine of rats given EEA. Dimethyldioxirane-mediated oxidation of EEA in the presence of NAL and GSH produced M1-M6, all of which were generated in microsomal incubations. The structures of M3 and M6 were confirmed by (1)H and (13)C NMR. These findings provide evidence for the metabolic activation of EEA to the corresponding cis-enedial intermediate both in vitro and in vivo. Ketoconazole inhibited the microsomal production of the cis-enedial, and P450 3A4 was found to be the primary enzyme involved in the bioactivation of EEA. PMID:26286065

  16. Preliminary Phytochemical Screening and In Vitro Antioxidant Activities of Parkinsonia aculeata Linn.

    PubMed Central

    Sharma, Sonia; Vig, Adarsh Pal

    2014-01-01

    Butanol and hexane leaves extracts of Parkinsonia aculeata L. (Fabaceae) were assessed for its antioxidant potential by in vitro methods. Phytochemical analysis and antioxidant activity of plant extracts were studied using different in vitro assays. UPLC analysis of extracts was carried out for the identification of chemical constituents. The total phenolic contents of the butanol and hexane leaf extract were 42?mgGAE/g and 34?mgGAE/g whereas flavonoid contents of these extracts were found to be 0.044?mgRE/g and 0.005?mgRE/g, respectively. Among both extracts, butanol extract shows maximum inhibition (%) of 93.88%, 80.02%, 52.06%, 94.68%, and 69.37% in DPPH, non-site-specific and site-specific, FTC, and TBA assays and absorbance of 0.852 and 0.522 in reducing power and CUPRAC assay at the highest concentration tested. The FRAP and TAC values of butanol extract were found to be 678??M Fe(II)/g and 36?mgAAE/100?mg. UPLC analysis of extracts revealed the presence of various polyphenols. The tested plant extracts were found to possess potent antioxidant and free radical scavenging activity which may be due to the presence of flavonoids and polyphenols. PMID:24822217

  17. Preliminary phytochemical screening and in vitro antioxidant activities of Parkinsonia aculeata Linn.

    PubMed

    Sharma, Sonia; Vig, Adarsh Pal

    2014-01-01

    Butanol and hexane leaves extracts of Parkinsonia aculeata L. (Fabaceae) were assessed for its antioxidant potential by in vitro methods. Phytochemical analysis and antioxidant activity of plant extracts were studied using different in vitro assays. UPLC analysis of extracts was carried out for the identification of chemical constituents. The total phenolic contents of the butanol and hexane leaf extract were 42 mgGAE/g and 34 mgGAE/g whereas flavonoid contents of these extracts were found to be 0.044 mgRE/g and 0.005 mgRE/g, respectively. Among both extracts, butanol extract shows maximum inhibition (%) of 93.88%, 80.02%, 52.06%, 94.68%, and 69.37% in DPPH, non-site-specific and site-specific, FTC, and TBA assays and absorbance of 0.852 and 0.522 in reducing power and CUPRAC assay at the highest concentration tested. The FRAP and TAC values of butanol extract were found to be 678 ?M Fe(II)/g and 36 mgAAE/100 mg. UPLC analysis of extracts revealed the presence of various polyphenols. The tested plant extracts were found to possess potent antioxidant and free radical scavenging activity which may be due to the presence of flavonoids and polyphenols. PMID:24822217

  18. In vitro anticancer activity of stachydrine isolated from Capparis decidua on prostate cancer cell lines.

    PubMed

    Rathee, Permender; Rathee, Dharmender; Rathee, Deepti; Rathee, Sushila

    2012-01-01

    In this article we report our work on the isolation, characterisation and evaluation of in vitro anticancer activity of stachydrine on solid tumour cells. The in vitro activity was assessed by MTT assay and propidium iodide (PI) staining. Further, an attempt was also made to check the effect of stachydrine on the invasion and metastasis of cancer cells by inhibiting the expression of chemokine receptors (CXCR3 and CXCR4). The influence of stachydrine on the gene expression of CXCR3 and CXCR4 at mRNA and protein levels was examined. Studies revealed a dose dependent decrease in expression of mRNA, and protein levels were observed in stachydrine-treated human prostate cancer cells (PC-3 and LNcaP) as detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The data therefore provides direct evidence for the role of stachydrine as a potent anti-metastatic agent, which can markedly inhibit the malignancy and invasive capacity of malignant cancer cells. PMID:21988653

  19. Microparticles containing propolis and metronidazole: in vitro characterization, release study and antimicrobial activity against periodontal pathogens.

    PubMed

    de Souza Ferreira, Sabrina Barbosa; de Assis Dias, Bruno Rafael; Obregón, Clarissa Silva; Gomes, Carla Carolina; de Araújo Pereira, Raphaela Regina; Ribeiro Godoy, Janine Silva; Estivalet Svidzinski, Terezinha Inez; Bruschi, Marcos Luciano

    2014-03-01

    Ethylcellulose microparticles containing metronidazole and propolis extractive solution were prepared and evaluated in vitro against periodontal pathogens. Scanning electron microscopy, particle size analysis, drug entrapment efficiency and drug release of microparticles were determined. The antimicrobial activity of microparticles was evaluated against microorganisms of periodontal importance (Enterococcus faecalis, Streptococcus pyogenes, Streptococcus mutans, Staphylococcus aureus, Klebsiella pneumoniae and Escherichia coli). It was obtained particles with regular morphology, mean diameter of 1.23?µm, and entrapment efficiency for propolis and metronidazole were 91.41% and 22.23%, respectively. In vitro release studies of propolis and metronidazole from microparticles showed prolonged drug release and controlled by Fickian diffusion. Both propolis and metronidazole displayed activity against the tested strains. Moreover, the results showed that the strains of E. faecalis, S. pyogenes and S. mutans were more susceptible to the propolis and E. faecalis to the metronidazole. It was also observed that the amount of metronidazole to inhibit the microorganism strains in the physical mixture with propolis was smaller than in the metronidazole alone, suggesting potentiation effect between propolis and metronidazole. These microparticles would be useful for developing intermediary or eventual dosage form to be administered into the periodontal pocket more easily and safely. PMID:23356908

  20. Activation of 2,4-diaminotoluene to proximate carcinogens in vitro, and assay of DNA adducts.

    PubMed

    Citro, G; Galati, R; Verdina, A; Marini, S; Zito, R; Giardina, B

    1993-03-01

    1. 2,4-Diaminotoluene, which yields adducts with DNA in vivo, has been studied for its ability to form adducts in vitro. Metabolic activation with rat liver post-mitochondrial supernatant gave 300 adducts/10(6) nucleotides in calf thymus single-stranded DNA, under defined experimental conditions. 2. 2,4-Diaminotoluene-modified DNA and deoxyhomopolymers showed characteristic u.v. absorption spectra, exhibiting hyperchromic effects at 235 and 220 nm, and hypochromic effect at 260 nm. The difference spectra between diamine-modified and untreated DNA, or deoxyhomopolymer, were very similar to the spectrum of 2,4-diaminotoluene alone. 3. 2,4-Diaminotoluene-modified DNA was assayed by ELISA with specific monoclonal antibodies directed against diamine-DNA adducts. Reactions with poly-d(A) or poly-d(A-T) gave no spectral modification, and immunochemical analysis showed that the diamine did not bind to these polynucleotides. On the other hand, in the case of poly-d(G) or poly-d(C-G), strong immunoreactions were observed, demonstrating that the guanine base is involved in the binding of the diamine to DNA. 4. Monoclonal antibodies directed against different diamine-DNA adducts have shown that 80% of the in vitro metabolic activation involves the para amino group of the aromatic diamine. PMID:8498094

  1. Selection of a T7 promoter mutant with enhanced in vitro activity by a novel multi-copy bead display approach for in vitro evolution.

    PubMed

    Paul, Siddhartha; Stang, Alexander; Lennartz, Klaus; Tenbusch, Matthias; Überla, Klaus

    2013-01-01

    In vitro evolution of nucleic acids and proteins is a powerful strategy to optimize their biological and physical properties. To select proteins with the desired phenotype from large gene libraries, the proteins need to be linked to the gene they are encoded by. To facilitate selection of the desired phenotype and isolation of the encoding DNA, a novel bead display approach was developed, in which each member of a library of beads is first linked to multiple copies of a clonal gene variant by emulsion polymerase chain reaction. Beads are transferred to a second emulsion for an in vitro transcription-translation reaction, in which the protein encoded by each bead's amplicon covalently binds to the bead present in the same picoliter reactor. The beads then contain multiple copies of a clonal gene variant and multiple molecules of the protein encoded by the bead's gene variant and serve as the unit of selection. As a proof of concept, we screened a randomized library of the T7 promoter for high expression levels by flow cytometry and identified a T7 promoter variant with an ~10-fold higher in vitro transcriptional activity, confirming that the multi-copy bead display approach can be efficiently applied to in vitro evolution. PMID:23074193

  2. Anticancer activity of litchi fruit pericarp extract against human breast cancer in vitro and in vivo

    SciTech Connect

    Wang Xiujie . E-mail: xiujiewang@yahoo.com; Yuan Shulan; Wang Jing; Lin Ping; Liu Guanjian; Lu Yanrong; Zhang Jie; Wang, Wendong; Wei Yuquan . E-mail: yuquanwei@mail.sc.cninfo.net

    2006-09-01

    Litchi fruit pericarp (LFP) extract contains significant amounts of polyphenolic compounds and exhibits powerful antioxidative activity against fat oxidation in vitro. The purpose of this study is to confirm the anticancer activity of LFP extract on human breast cancer in vitro and in vivo, and to elucidate the mechanism of its activity. Human breast cancer cells were tested in vitro for cytotoxicity, colony formation inhibition, BrdU incorporation, and gene expression profiling after treatment with LFP extract. Seven nude mice bearing human breast infiltrating duct carcinoma orthotopically were tested for its anticancer activity and expression of caspase-3 in vivo by oral administration of 0.3% (0.3 mg/ml) of LFP water-soluble crude ethanolic extract (CEE) for 10 weeks. LFP extract demonstrated a dose- and time-dependent inhibitory effect on cell growth (IC{sub 5} = 80 {mu}g/ml), and it significantly inhibited colony formation and BrdU incorporation of human breast cancer cells. Oligonucleotide microarray analysis identified 41(1.22%) up-regulated and 129 (3.84%) down-regulated genes after LFP water-soluble CEE treatment; the predominantly up-regulated genes were involved in various biological functions including cell cycle regulation and cell proliferation, apoptosis, signal transduction and transcriptional regulation, and extracellular matrix/adhesion molecules; and down-regulated genes were mainly associated with adhesion, invasion, and malignancy of cancer cells. A 40.70% tumor mass volume reduction and significant increase of casepase-3 protein expression were observed in vivo experiment. The findings in this study suggested that LFP extract might have potential anticancer activity on both ER positive and negative breast cancers, which could be attributed, in part, to its DNA damage effect, proliferating inhibition and apoptosis induction of cancer cells through up-regulation and down-regulation of multiple genes involved in cell cycle regulation and cell proliferation, apoptosis, signal transduction and transcriptional regulation, motility and invasiveness of cancer cells; ADP-ribosyltransferase (NAD+; poly (ADP-ribose) polymerase)-like 1 (ADPRTL1), Cytochrome P450, subfamily I (CYP1A1) and Hyaluronan-mediated motility receptor (HMMR) might be the main molecular targets at which LFP water-soluble CEE acted.

  3. Anti-tumor activity and cytotoxicity in vitro of novel 4,5-dialkylimidazolium surfactants.

    PubMed

    Wang, Da; Richter, Christian; Rühling, Andreas; Hüwel, Sabine; Glorius, Frank; Galla, Hans-Joachim

    2015-11-27

    Natural monoalkylated imidazolium derivatives exhibit significant anti-tumor activity as well as general cytotoxicity. In the present study, we used a series of newly synthesized imidazolium derivatives bearing two alkyl chains in the backbone of the imidazolium core in 4- and 5-position and either dimethyl- or dibenzyl-substituents at 1- and 3-position. Their anti-tumor activity and cytotoxicity were determined in vitro using the lactate dehydrogenase (LDH) assay. The tumor cell line C6 from rat glioma, the non-tumor MDCK cell line (Madin-Darby canine kidney) as well as the mouse embryonic fibroblast cell line (NIH3T3) were used as cellular targets. Surface activity measurements were performed leading to the determination of their critical micelle concentration (CMC) of these new lipid analogues to evaluate the molecular mechanism of the observed cellular effects. We found that 4,5-dialkylation of the imidazole ring enhances the anti-tumor activity compared to simple 1-alkylated imidazoles. The corresponding C7 homologues are found to be the most potent compounds. Furthermore dibenzyl-substituted imidazolium surfactants exhibit higher surface activity and increased toxicity against tumor cells compared to dimethyl-substituted imidazolium surfactants. In summary the dibenzyl-derivative carrying the two C7 chains was found to exhibit a drastically increased anti-tumor activity especially compared to so far known monoalkylated species. PMID:26456641

  4. An in vitro screening with emerging contaminants reveals inhibition of carboxylesterase activity in aquatic organisms.

    PubMed

    Solé, Montserrat; Sanchez-Hernandez, Juan C

    2015-12-01

    Pharmaceuticals and personal care products (PPCPs) form part of the new generation of pollutants present in many freshwater and marine ecosystems. Although environmental concentrations of these bioactive substances are low, they cause sublethal effects (e.g., enzyme inhibition) in non-target organisms. However, little is known on metabolism of PPCPs by non-mammal species. Herein, an in vitro enzyme trial was performed to explore sensitivity of carboxylesterase (CE) activity of aquatic organisms to fourteen PPCPs. The esterase activity was determined in the liver of Mediterranean freshwater fish (Barbus meridionalis and Squalius laietanus), coastal marine fish (Dicentrarchus labrax and Solea solea), middle-slope fish (Trachyrhynchus scabrus), deep-sea fish (Alepocephalus rostratus and Cataetix laticeps), and in the digestive gland of a decapod crustacean (Aristeus antennatus). Results showed that 100?M of the lipid regulators simvastatin and fenofibrate significantly inhibited (30-80% of controls) the CE activity of all target species. Among the personal care products, nonylphenol and triclosan were strong esterase inhibitors in most species (36-68% of controls). Comparison with literature data suggests that fish CE activity is as sensitive to inhibition by some PPCPs as that of mammals, although their basal activity levels are lower than in mammals. Pending further studies on the interaction between PPCPs and CE activity, we postulate that this enzyme may act as a molecular sink for certain PPCPs in a comparable way than that described for the organophosphorus pesticides. PMID:26562051

  5. Xenobiotic metabolizing enzyme activities in cells used for testing skin sensitization in vitro.

    PubMed

    Fabian, E; Vogel, D; Blatz, V; Ramirez, T; Kolle, S; Eltze, T; van Ravenzwaay, B; Oesch, F; Landsiedel, R

    2013-09-01

    For ethical and regulatory reasons, in vitro tests for scoring potential toxicities of cosmetics are essential. A test strategy for investigating potential skin sensitization using two human keratinocytic and two human dendritic cell lines has been developed (Mehling et al. Arch Toxicol 86:1273–1295, 2012). Since prohaptens may be metabolically activated in the skin, information on xenobiotic metabolizing enzyme (XME) activities in these cell lines is of high interest. In this study, XME activity assays, monitoring metabolite or cofactor, showed the following: all three passages of keratinocytic (KeratinoSens® and LuSens) and dendritic (U937 und THP-1) cells displayed N-acetyltransferase 1 (NAT1) activities (about 6–60 nmol/min/mg S9-protein for acetylation of para-aminobenzoic acid). This is relevant since reactive species of many cosmetics are metabolically controlled by cutaneous NAT1. Esterase activities of about 1–4 nmol fluorescein diacetate/min/mg S9-protein were observed in all passages of investigated keratinocytic and about 1 nmol fluorescein diacetate/min/mg S9-protein in dendritic cell lines. This is also of practical relevance since many esters and amides are detoxified and others activated by cutaneous esterases. In both keratinocytic cell lines, activities of aldehyde dehydrogenase (ALDH) were observed (5–17 nmol product/min/mg cytosolic protein). ALDH is relevant for the detoxication of reactive aldehydes. Activities of several other XME were below detection, namely the investigated cytochrome P450-dependent alkylresorufin O-dealkylases 7-ethylresorufin O-deethylase, 7-benzylresorufin O-debenzylase and 7-pentylresorufin O-depentylase (while NADPH cytochrome c reductase activities were much above the limit of quantification), the flavin-containing monooxygenase, the alcohol dehydrogenase as well as the UDP glucuronosyl transferase activities. PMID:23958860

  6. In vitro Activation of heme oxygenase-2 by menadione and its analogs

    PubMed Central

    2014-01-01

    Background Previously, we reported that menadione activated rat, native heme oxygenase-2 (HO-2) and human recombinant heme oxygenase-2 selectively; it did not activate spleen, microsomal heme oxygenase-1. The purpose of this study was to explore some structure–activity relationships of this activation and the idea that redox properties may be an important aspect of menadione efficacy. Methods Heme oxygenase activity was determined in vitro using rat spleen and brain microsomes as the sources of heme oxygenase-1 and ?2, respectively, as well as recombinant, human heme oxygenase-2. Results Menadione analogs with bulky aliphatic groups at position-3, namely vitamins K1 and K2, were not able to activate HO-2. In contrast, several compounds with similar bulky but less lipophilic moieties at position-2 (and ?3) were able to activate HO-2 many fold; these compounds included polar, rigid, furan-containing naphthoquinones, furan-benzoxazine naphthoquinones, 2-(aminophenylphenyl)-3-piperidin-1-yl naphthoquinones. To explore the idea that redox properties might be involved in menadione efficacy, we tested analogs such as 1,4-dimethoxy-2-methylnaphthalene, pentafluoromenadione, monohalogenated naphthoquinones, ?-tetralone and 1,4-naphthoquinone. All of these compounds were inactive except for 1,4-naphthoquinone. Menadione activated full-length recombinant human heme oxygenase-2 (FL-hHO-2) as effectively as rat brain enzyme, but it did not activate rat spleen heme oxygenase. Conclusions These observations are consistent with the idea that naphthoquinones such as menadione bind to a receptor in HO-2 and activate the enzyme through a mechanism that may involve redox properties. PMID:24533775

  7. Inhibitory Activities of Cudrania tricuspidata Leaves on Pancreatic Lipase In Vitro and Lipolysis In Vivo

    PubMed Central

    Kim, Young Sook; Lee, Youngseop; Kim, Junghyun; Sohn, Eunjin; Kim, Chan Sik; Lee, Yun Mi; Jo, Kyuhyung; Shin, Sodam; Song, Yoojin; Kim, Joo Hwan; Kim, Jin Sook

    2012-01-01

    To identify effective herb to treat obesity, we screened 115 herbal extracts for inhibition of porcine pancreatic lipase (triacylg-ycerol acylhydrolase, EC 3.1.1.3) activity in vitro. Of the extracts tested, Cudrania tricuspidata leaves exhibited the most pronounced inhibitory effect on lipase activity with an IC50 value of 9.91??g/mL. Antilipid absorption effects of C. tricuspidata leaves were examined in rats after oral administration of lipid emulsions containing 50 or 250?mg??C. tricuspidata/kg body weight. Plasma triacylglycerol levels 2?h after the oral administration of emulsions containing C. tricuspidata were significantly reduced compared to the untreated group (P < 0.05). These results suggest that C. tricuspidata leaves may be useful for the treatment of obesity. PMID:23365603

  8. Partial in vitro analysis of toxic and antigenic activities of eleven Peruvian pitviper snake venoms.

    PubMed

    Guerra-Duarte, C; Lopes-Peixoto, J; Fonseca-de-Souza, B R; Stransky, S; Oliveira, D; Schneider, F S; Lopes-de-Souza, L; Bonilla, C; Silva, W; Tintaya, B; Yarleque, A; Chávez-Olórtegui, C

    2015-12-15

    This work used eleven Peruvian snake venoms (Bothrops andianus, Bothrops atrox, Bothrops barnetti, Bothrops castelnaudi, Bothriopsis chloromelas, Bothrocophias microphthalmus, Bothrops neuwiedi, Bothriopsis oligolepis, Bothriopsis peruviana, Bothrops pictus and Bothriopsis taeniata) to perform in vitro experimentation and determine its main characteristics. Hyaluronidase (HYAL), phospholipase A2 (PLA2), snake venom metalloproteinase (SVMP), snake venom serine protease (SVSP) and l-amino acid oxidase (LAAO) activities; toxicity by cell viability assays using MGSO3, VERO and HeLa cell lineages; and crossed immunoreactivity with Peruvian (PAV) and Brazilian (BAV) antibothropic polyvalent antivenoms, through ELISA and Western Blotting assays, were determined. Results show that the activities tested in this study were not similar amongst the venoms and each species present their own peculiarities, highlighting the diversity within Bothrops complex. All venoms were capable of reducing cell viability of all tested lineages. It was also demonstrated the crossed recognition of all tested venoms by both antivenoms. PMID:26365916

  9. In vitro activities of a wide panel of antifungal drugs against various Scopulariopsis and Microascus species.

    PubMed

    Skóra, Magdalena; Bulanda, Ma?gorzata; Jagielski, Tomasz

    2015-09-01

    The in vitro activities of 11 antifungal drugs against 68 Scopulariopsis and Microascus strains were investigated. Amphotericin B, 5-fluorocytosine, fluconazole, itraconazole, ketoconazole, miconazole, posaconazole, voriconazole, and ciclopirox showed no or poor antifungal effect. The best activities were exhibited by terbinafine and caspofungin, where the MIC and MEC (minimal effective concentration) ranges were 0.0313 to >16 ?g/ml and 0.125 to 16 ?g/ml, respectively. The MIC and MEC modes were both 1 µg/ml for terbinafine and caspofungin; the MIC50 and MEC50 were 1 µg/ml for both drugs, whereas the MIC90 and MEC90 were 4 µg/ml and 16 µg/ml, respectively. PMID:26100698

  10. An in vitro study of electrically active hydroxyapatite-barium titanate ceramics using Saos-2 cells.

    PubMed

    Baxter, Frances R; Turner, Irene G; Bowen, Christopher R; Gittings, Jonathan P; Chaudhuri, Julian B

    2009-08-01

    Electrically active ceramics are of interest as bone graft substitute materials. This study investigated the ferroelectric properties of hydroxyapatite-barium titanate (HABT) composites and the behaviour of osteoblast-like cells seeded on their surfaces. A piezoelectric coefficient (d(33)) of 57.8 pCN(-1) was observed in HABT discs prepared for cell culture. The attachment, proliferation, viability, morphology and metabolic activity of cells cultured on unpoled HABT were comparable to those observed on commercially available hydroxyapatite at all time points. No indication of the cytotoxicity of HABT was detected. At one day after seeding, cell attachment was modified on both the positive and negative surfaces of poled HABT. After longer incubations, all parameters observed were comparable on poled and unpoled ceramics. The results indicate that HABT ceramics are biocompatible in the short term in vitro and that further investigation of cell responses to these materials under mechanical load and at longer incubation times is warranted. PMID:19308338

  11. Optimization of ultrasonic-assisted extraction and in vitro antioxidant activities of polysaccharides from Trametes orientalis.

    PubMed

    Zheng, Yi; Li, Yong; Wang, Wei-dong

    2014-10-13

    A Box-Behnken design was employed to optimize ultrasonic-assisted extraction of Trametes orientalis polysaccharides (TOP). The crude polysaccharides were purified by DEAE cellulose-52 chromatography, giving a main fraction named as PTOP. The antioxidant properties of PTOP were evaluated by different in vitro antioxidant assays, such as 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities, reducing power, superoxide radical scavenging activities, and chelating ability of ferrous ions. The results showed that optimal extraction parameters were as follows: ratio of water to raw material 30.6 mL/g, ultrasonic power 109.8 W, extraction temperature 40.2 °C, and extraction time 42.2 min. Under these conditions, the experimental yield of polysaccharides was 7.49 ± 0.14%, which agreed closely with the predicted value (7.47%). Furthermore, PTOP exhibited antioxidant capacity in a concentration-dependent manner in all assays. PMID:25037357

  12. In vitro biological activities and fatty acid profiles of Pistacia terebinthus fruits and Pistacia khinjuk seeds.

    PubMed

    Hac?bekiro?lu, I?il; Y?lmaz, Pelin Köseo?lu; Ha?imi, Nesrin; K?l?nç, Ersin; Tolan, Veysel; Kolak, Ufuk

    2015-01-01

    This study reports in vitro anticholinesterase, antioxidant and antimicrobial effects of the n-hexane, dichloromethane, ethanol and ethanol-water extracts prepared from Pistacia terebinthus L. fruits and Pistacia khinjuk Stocks seeds as well as their total phenolic and flavonoid contents, and fatty acid compositions. Ethanol and ethanol-water extracts of both species exhibited higher anticholinesterase activity than galanthamine. Among ABTS, DPPH and CUPRAC assays, the highest antioxidant capacity of the extracts was found in the last one. P. terebinthus ethanol extract being rich in flavonoid content showed the best cupric reducing effect. All extracts possessed no antimicrobial activity. The main fatty acid in P. terebinthus fruits (52.52%) and P. khinjuk seeds (59.44%) was found to be oleic acid. Our results indicate that P. terebinthus fruits and P. khinjuk seeds could be a good source of anticholinesterase compounds, and could be phytochemically investigated. PMID:25115646

  13. In vitro and in vivo antioxidant activity of polyphenols extracted from black highland barley.

    PubMed

    Shen, Yingbin; Zhang, Hui; Cheng, Liling; Wang, Li; Qian, Haifeng; Qi, Xiguang

    2016-03-01

    The objective of this study was to determine the antioxidant capacity of polyphenols extracted from black highland barley (BHLPE) in vitro and in vivo. BHLPE was found to have strong superoxide radical, hydroxyl radical and 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity, ferric reducing antioxidant power and moderate metal ion-chelating activity. Compared with a high fat diet (HFD) group, mice that were administered 600mg BHLPE/kg body weight showed significant decreases in total cholesterol, low-density lipoprotein cholesterol and the atherosclerosis index, in addition to markedly increased high-density lipoprotein cholesterol levels. Furthermore, the antioxidant defense system and antioxidant gene expression were significantly improved in vivo in mice that were administered BHLPE compared with mice in the HFD group. These results suggest that BHLPE has significant potential as a natural antioxidant to promote health and to reduce the risk of disease. PMID:26471646

  14. Antibacterial activity of aqueous extracts of Indian chewing sticks on dental plaque: An in vitro study

    PubMed Central

    Rao, Dola Srinivasa; Penmatsa, Tanuja; Kumar, Alapati Kranthi; Reddy, M. Narendra; Gautam, Nalam Sai; Gautam, Nalam Radhika

    2014-01-01

    The anti-microbial efficacy of aqueous extracts of Indian chewing sticks against different kinds of plaque bacteria in vitro was investigated. Supra-gingival plaque is cultured and subjected to the antibacterial activity of the aqueous extracts of chewing sticks (Neem, Acacia, Pongamia glabra, Achyranthes aspera, Streblus asper) separately. The results of the study demonstrate that all the five chewing sticks under study possess inhibitory potential against bacteria present in dental plaque mainly on aerobes. The antibacterial efficacy of aqueous extracts has antibacterial effects and could be used as a therapeutic agent and therefore, it appears to be potent anti-microbial agents that could be considered as a medicinal plant. Results of this study showed chewing sticks contained antibacterial agents, but the concentration and composition of the active substances differed among the plants. PMID:25210357

  15. Chemical interaction of disulfiram with nitrosodimethylamine after in vitro enzymatic activation

    SciTech Connect

    Tacchi, A.M.; Bertram, B.; Wiessler, M.

    1984-02-01

    The in vitro reaction between disulfiram (DSF) and N-nitroso(/sup 14/C)dimethylamine ((/sup 14/C)NDMA) was studied. Incubations of DSF with (/sup 14/C)NDMA were carried out in the presence of rat liver microsomes, control 9000 g (S9) supernatant fraction and phenobarbital-induced S9 fraction. HPLC analysis and liquid scintillation measurement provided evidence for the formation of methyldiethyldithiocarbamate (MeDDTC) as a product of the reaction between diethyldithiocarbamate (DDTC), the main active metabolite of DSF and the 'methyl-cation' released by NDMA after enzymatic activation. The amount of MeDDTC found here was consistent with the rate of oxidation of NDMA to formaldehyde. Scintillation counting confirmed that other radioactive peaks, not due to MeDDTC, were unrelated to the methylation of L-cysteine by (/sup 14/C)NDMA.

  16. Inhibition of iron induced lipid peroxidation and antioxidant activity of Indian spices and Acacia in vitro.

    PubMed

    Yadav, Amit Singh; Bhatnagar, Deepak

    2010-03-01

    The spices used in the Indian foods such as Star anise (Illicium verum), Bay leaves (Cinnamomum zeylanicum) and Cobra's saffron (Mesua ferrea), and Acacia (Acacia catechu), which have medicinal value, were used as test samples, to find their effect on in vitro lipid peroxidation (LPO). Rat liver post mitochondrial supernatant (PMS) in Tris HCl buffer, pH 7.4 was incubated for 0 and 1 h, with various test extracts in three different oxidant systems. The results show that addition of test samples to FeCl(3) medium at 0 h significantly stop the initiation of the LPO. However, the propagation phase of LPO was inhibited by Cobra's saffron and Acacia and not by Star anise and Bay leaves. The test samples also showed strong reducing power and superoxide radical scavenging activity. Cobra's saffron and Acacia showed the highest antioxidant activity, probably due to the higher polyphenol content as compared to other test samples. PMID:20033297

  17. Synthesis, characterization, in vitro anti-proliferative and hemolytic activity of hydroxyapatite

    NASA Astrophysics Data System (ADS)

    Palanivelu, R.; Ruban Kumar, A.

    2014-06-01

    Hydroxyapatite (Ca10(PO4)6(OH)2, HAP) nanoparticles are widely used in several biomedical applications due to its compositional similarities to bone mineral, excellent biocompatibility and bioactivity, osteoconductivity. In this present investigation, HAP nanoparticles synthesized by precipitation technique using calcium nitrate and di-ammonium phosphate. The crystalline nature and the functional group analysis are confirmed using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and Fourier transform Raman spectroscopy (FT-Raman) respectively. The morphological observations are ascertained from field emission electron scanning electron microscope (FE-SEM) and transmission electron microscope (TEM). In vitro anti-proliferative and hemolytic activities are carried out on the synthesized HAP samples and the studies reveals that HAP have mild activity against erythrocytes.

  18. Evaluation of antimicrobial activity of extracts of in vivo and in vitro grown Vinca rosea L. (Catharanthus roseus) against pathogens.

    PubMed

    Naz, Shagufta; Haq, Rukhama; Aslam, Farah; Ilyas, Saiqa

    2015-05-01

    The antimicrobial activity of Vinca rosea was evaluated against pathogenic bacterial strains (Bacillus subtilis, B. licheniformis and Azotobacter sp.) and fungal strains (Asprgillus niger, Alternaria solani and Rhizopus oryzae) using agar well diffusion method. Methanolic extracts of in vivo leaf, in vitro leaf, in vitro calluses of leaf, nodal and fruit explants were used and exhibited antimicrobial activity as indicated by minimum inhibitory concentration (MIC). In vitro extracts showed better results as compared to the in vivo extracts for both the antibacterial as well as the antifungal activity. Among all the extracts, maximum zone of inhibition (30.3 mm ± 0.58(a)) was formed by in vitro leaf callus extract concentration of 2.0mg/ml against B. licheniformis. Similarly in case of antifungal activity, maximum zone of inhibition (34.6mm ± 0.57(a)) was formed by in vitro leaf callus extract and MIC value is 6.0mg/ml against A. niger. Hence these results clearly depicts that V. rosea possess a great strength to fight against the microbial activity and can be used against various infections. PMID:26004716

  19. Activation of VEGF and FGF induced angiogenesis under influence of low level laser radiation in vitro

    NASA Astrophysics Data System (ADS)

    Gasparyan, Levon; Brill, Grigory; Makela, Anu

    2006-02-01

    One of the feasible explanations for long-term treatment effects of laser therapy of diseases connected with tissue ischemia and altered blood circulation is activation of angiogenesis after low level laser irradiation. The aim of the current study was to investigate if laser irradiation can enhance vascular endothelial growth factor (VEGF) or basic fibroblast growth factor (FGF) induced angiogenesis in vitro. The study was conducted on rat thoracic aortal rings. Samples of group 1 served as control, samples of groups 2 and 3 were incubated with VEGF or FGF, group 4 samples were irradiated with laser (660 nm, 20 mW) during 10 min, samples of groups 5 and 6 were incubated with VEGF or FGF accordingly and received 10 min of laser irradiation. In the control group no noticeable angiogenesis occurred. The application of VEGF activated angiogenesis: the area covered by new vessels was 1,3+/-0,24 mm2 and the maximal length of vessels was 0,93+/-0,11 mm. Laser light irradiation (group 4) activated angiogenesis (1,9+/-0,29 mm2 and 0,75+/-0,10 mm). The combined influence of laser light and VEGF on angiogenesis (group 5) was significantly stronger (p <0,001), than each of the factors separately (6,98+/-0,88 mm2 and 1,7+/-0,23 mm). Application of FGF also activated angiogenesis: the area covered by new vessels was 2,76+/-0,22 mm2 and the maximal length of vessels was 1,19+/-0,12 mm. Combined influence of laser light and FGF on angiogenesis (group 6) was again significantly stronger (p <0,001), than each of the factors separately (5,43+/-0,28 mm2 and 1,99+/-0,10 mm). Studies show that laser irradiation can intensify effects of growth factors in vitro.

  20. In vitro and in vivo antidermatophytic activities of some Iranian medicinal plants.

    PubMed

    Ayatollahi Mousavi, Seyyed Amin; Kazemi, Abdolhasan

    2015-11-01

    In the last decades, the number of people suffering from dermatophytoses has seriously increased, which may be due to the development of resistant strains to a range of antifungal drugs. The present study was aimed to evaluate the antidermatophytic properties of eight extracts from the selected spices and herbs, which were ethno-medicinally used in Iran against Trichophyton mentagrophytes, Trichophyton interdigitale, Microsporum canis, and Microsporum gypseum (10 strain of each). The in vitro antifungal activities of the extracts from four spices and four plants were evaluated by the broth macro dilution method against four dermatophyte strains. In addition, the in vivo therapeutic effects of Myrtus communis L. and Cinnamomum zeylanicum Blume extracts (the most active extracts) on dermatophytosis induced by M. canis and T. mentagrophytes in guinea pigs were evaluated. Results of in vitro antifungal assay revealed that all the tested extracts demonstrated both fungistatic and fungicidal activities with the geometric mean (GM) MIC ranging from 0.058 to 3.73?mg/ml and GM (MFC) ranging from 0.058 to 7.46?mg/ml, respectively. Two extracts (M. communis and C. zeylanicum) significantly inhibited the growth of all the tested dermatophytes, while other extracts demonstrated weak (MICs of >0.625?mg/ml) to moderate (MICs ranging from 100 to 0.625?mg/ml) activities. In vivo antidermatophytic assay demonstrated that clotrimazole cured T. mentagrophytes and M. canis infection on days 21 and 17, respectively, whereas M. communis and C. zeylanicum extracts significantly (p < 0.05) cured T. mentagrophytes and M. canis infection on days 9 and 13 as well as 9, 11, respectively. Phytochemical screening showed the presence of flavonoids, tannins, phenols, and alkaloids in M. communis and alkaloids, flavonoids, and tannins in C. zeylanicum. Findings of the present study also provided the scientific evidence that natural plants could be used in traditional medicine for the prevention and treatment of dermatophytic infections. PMID:26092105

  1. Using an In Vitro Cell Line to Assess Source and Treated Drinking Water Extracts for Estrogenic Activity.

    EPA Science Inventory

    While in vitro assays have been used to determine presence of estrogenic activity in many types of water, few studies have evaluated the potential estrogenic activity in source and treated drinking water samples. In a collaborative research study the U.S. Environmental Protection...

  2. Escherichia coli RecX Inhibits RecA Recombinase and Coprotease Activities in Vitro and in Vivo*

    E-print Network

    Kowalczykowski, Stephen C.

    Escherichia coli RecX Inhibits RecA Recombinase and Coprotease Activities in Vitro and in Vivo and Development, University of California, Davis, California 95616 In Escherichia coli the RecA protein playsA recombinase and coprotease activities. The Escherichia coli RecA protein plays a central role in homologous

  3. Structure-activity relationships for in vitro diuretic activity of CAP2b in the housefly

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A series of truncated and Ala-replacement analogs of the peptide Manse-CAP2b (pELYAFPRV-NH2) were assayed for diuretic activity on Malpighian tubules of the housefly Musca domestica. The C-terminal hexapeptide proved to be the active core, the minimum sequence required to retain significant diureti...

  4. Bromelain treatment reduces CD25 expression on activated CD4+ T cells in vitro?

    PubMed Central

    Secor, Eric R.; Singh, Anurag; Guernsey, Linda A.; McNamara, Jeff T.; Zhan, Lijun; Maulik, Nilanjana; Thrall, Roger S.

    2009-01-01

    Bromelain (Br), an extract from pineapple stem with cysteine protease activity, exerts anti-inflammatory effects in a number of inflammatory models. We have previously shown that Br treatment decreased activated CD4+ T cells and has a therapeutic role in an ovalbumin-induced murine model of allergic airway disease. The current study was designed to determine the effect of Br on CD4+ T cell activation, specifically the expression of CD25 in vitro. CD25 is up regulated upon T cell activation, found as a soluble fraction (sCD25) and is a therapeutic target in inflammation, autoimmunity and allergy. Br treatment of anti-CD3 stimulated CD4+ T cells reduced CD25 expression in a dose and time dependent manner. This reduction of CD25 was dependent on the proteolytic action of Br as the addition of E64 (a cysteine protease inhibitor) abrogated this response. The concentration of sCD25 was increased in supernatants of Br treated activated CD4+ T cells as compared to control cells, suggesting that Br proteolytically cleaved cell-surface CD25. This novel mechanism of action identifies how Br may exert its therapeutic benefits in inflammatory conditions. PMID:19162239

  5. Screening for hemostatic activities of popular Chinese medicinal herbs in vitro

    PubMed Central

    Ohkura, Naoki; Yokouchi, Haruna; Mimura, Mariyo; Nakamura, Riki; Atsumi, Gen-ichi

    2015-01-01

    Aims: This study aimed to identify new hemostyptics by assessing the coagulation enhancing activity of 114 Chinese herbal extracts in vitro. Methods: Herbs were boiled in water for 30 min, filtered and then lyophilized filtrates (10 mg/mL) were dissolved in water. Coagulation was assayed as prothrombin time (PT). Plasma diluted in saline was incubated with each extract for 5 min and then PT reagent was added, followed by CaCl2 solution and the time taken to form clots was measured. Extracts that decreased coagulation time were regarded as containing active compounds. The abilities of extracts to activate Factor XII were assessed and the activated form of factor XII (XIIa) was resolved by SDS-PAGE and visualized by silver staining. Results: Coagulation time was obviously shortened by extracts of Alpinia Rhizome, Areca, Artemisia Leaf, Cassia Bark, Danshen Root, Ephedra Herb, Epimedium Herb, Forsythia Fruit, Great Burdock Achene, Moutan Bark, Perilla Herb, Red Paeony Root, Schizonepeta Spike, Senticosus Rhizome, Sweet Annie, Uncaria Thorn and Zanthoxylum Peel. Factor XII was obviously activated by extracts of Artemisia Leaf and Great Burdock Achene, and slightly by Perilla herb. Conclusion: Some popular Chinese medicinal herbs have potential as hemostatic agents and could thus be develope as new strategies for the treatment and prevention of bleeding. PMID:26401379

  6. Catalytic diesel particulate filters reduce the in vitro estrogenic activity of diesel exhaust.

    PubMed

    Wenger, Daniela; Gerecke, Andreas C; Heeb, Norbert V; Naegeli, Hanspeter; Zenobi, Renato

    2008-04-01

    An in vitro reporter gene assay based on human breast cancer T47D cells (ER-CALUX) was applied to examine the ability of diesel exhaust to induce or inhibit estrogen receptor (ER)-mediated gene expression. Exhaust from a heavy-duty diesel engine was either treated by iron- or copper/iron-catalyzed diesel particulate filters (DPFs) or studied as unfiltered exhaust. Collected samples included particle-bound and semivolatile constituents of diesel exhaust. Our findings show that all of the samples contained compounds that were able to induce ER-mediated gene expression as well as compounds that suppressed the activity of the endogenous hormone 17beta-estradiol (E2). Estrogenic activity prevailed over antiestrogenic activity. We found an overall ER-mediated activity of 1.63 +/- 0.31 ng E2 CALUX equivalents (E2-CEQs) per m(3) of unfiltered exhaust. In filtered exhaust, we measured 0.74 +/- 0.07 (iron-catalyzed DPF) and 0.55 +/- 0.09 ng E2-CEQ m(-3) (copper/iron-catalyzed DPF), corresponding to reductions in estrogenic activity of 55 and 66%, respectively. Our study demonstrates that both catalytic DPFs lowered the ER-mediated endocrine-disrupting potential of diesel exhaust. PMID:18264702

  7. In vitro antimicrobial activity and antagonistic effect of essential oils from plant species.

    PubMed

    Toroglu, Sevil

    2007-07-01

    Kahramanmaras, is a developing city located in the southern part of Turkey Thymus eigii (M. Zohary and RH. Davis) Jalas, Pinus nigraAm. sub sp pallasiana and Cupressus sempervirens L. are the useful plants of the Kahramanmaras province and have been understudy since 2004 for the traditional uses of plants empiric drug, spice, herbal tea industry herbal gum and fuel. The study was designed to examine the antimicrobial activities of essential oils of these plants by the disc diffusion and minimum inhibitory concentration (MIC) methods. In addition, antimicrobial activity of Thymus eigii was researched by effects when it was used together with antibiotics and even when it was combined with other essential oils. When the results of this study were compared with vancomycin (30 mcg) and erytromycin (15 mcg) standards, it was found that Thymus eigii essential oil was particularly found to possess strongerantimicrobial activity whereas other essential oils showed susceptible or moderate activity However, antimicrobial activity changed also by in vitro interactions between antibiotics and Thymus eigii essential oil, also between essential oils of these plants and that of Thymus eigii causing synergic, additive, antagonist effect. PMID:18380074

  8. In vitro antiplasmodial activity of benzophenones and xanthones from edible fruits of Garcinia species.

    PubMed

    Lyles, James T; Negrin, Adam; Khan, Shabana I; He, Kan; Kennelly, Edward J

    2014-06-01

    Species of Garcinia have been used to combat malaria in traditional African and Asian medicines, including Ayurveda. In the current study, we have identified antiplasmodial benzophenone and xanthone compounds from edible Garcinia species by testing for in vitro inhibitory activity against Plasmodium falciparum. Whole fruits of Garcinia xanthochymus, G. mangostana, G. spicata, and G. livingstonei were extracted and tested for antiplasmodial activity. Garcinia xanthochymus was subjected to bioactivity-guided fractionation to identify active partitions. Purified benzophenones (1-9) and xanthones (10-18) were then screened in the plasmodial lactate dehydrogenase assay and tested for cytotoxicity against mammalian (Vero) cells. The benzophenones guttiferone E (4), isoxanthochymol (5), and guttiferone H (6), isolated from G. xanthochymus, and the xanthones ?-mangostin (15), ?-mangostin (16), and 3-isomangostin (17), known from G. mangostana, showed antiplasmodial activity with IC50 values in the range of 4.71-11.40 µM. Artemisinin and chloroquine were used as positive controls and exhibited IC50 values in the range of 0.01-0.24 µM. The identification of antiplasmodial benzophenone and xanthone compounds from G. xanthochymus and G. mangostana provides evidence for the antiplasmodial activity of Garcinia species and warrants further investigation of these fruits as dietary sources of chemopreventive compounds. PMID:24963617

  9. Mitogenic activity for fibroblasts induced by silica and titanium dioxide particles in vitro and in vivo.

    PubMed Central

    Kumar, R. K.; O'Grady, R.; Li, W.; Velan, G. M.

    1992-01-01

    Experimental studies on particle-induced pulmonary fibrosis have not provided consistent evidence for the specific induction of fibroblast-regulating cytokines by pulmonary macrophages in response to fibrogenic as compared to non-fibrogenic particles. Using an optimized, wholly serum-free bioassay, we assessed mitogenic activity for pulmonary fibroblasts in supernatants of short-term cultures of alveolar macrophages exposed to either fibrogenic silica or non-fibrogenic titanium dioxide ducts. The responses to these supernatants were influenced by the replicative status of the target cells, in that samples which stimulated non-cycling fibroblasts caused inhibition of DNA synthesis by cycling cells when tested at the same concentration. However, both silica and titanium dioxide elicited comparable secretion of growth factor activity by macrophages, following either in-vitro or in-vivo administration of particles. In contrast, bronchoalveolar lavage fluids from animals that received intratracheal injections of silica, but not from those that received titanium dioxide, exhibited a sustained reduction in fibroblast-stimulating activity. We conclude that secretion of growth factor activity by alveolar macrophages in culture is induced by particles in a non-specific manner. However, alterations in mitogenic activity in bronchoalveolar lavage fluid may constitute a biological marker of the pattern of pulmonary injury which progresses to fibrosis. PMID:1329913

  10. Mutational analysis of the Tetrahymena telomerase RNA: identification of residues affecting telomerase activity in vitro.

    PubMed

    Autexier, C; Greider, C W

    1998-02-01

    Telomere-specific repeat sequences are essential for chromosome end stability. Telomerase maintains telomere length by adding sequences de novo onto chromosome ends. The template domain of the telomerase RNA component dictates synthesis of species-specific telomeric repeats and other regions of the RNA have been suggested to be important for enzyme structure and/or catalysis. Using enzyme reconstituted in vitro with RNAs containing deletions or substitutions we identified nucleotides in the RNA component that are important for telomerase activity. Although many changes to conserved features in the RNA secondary structure did not abolish enzyme activity, levels of activity were often greatly reduced, suggesting that regions other than the template play a role in telomerase function. The template boundary was only altered by changes in stem II that affected the conserved region upstream of the template, not by changes in other regions, such as stems I, III and IV, consistent with a role of the conserved region in defining the 5' boundary of the template. Surprisingly, telomerase RNAs with substitutions or deletion of residues potentially abolishing the conserved pseudoknot structure had wild-type levels of telomerase activity. This suggests that this base pairing interaction may not be required for telomerase activity per se but may be conserved as a regulatory site for the enzyme in vivo. PMID:9443971

  11. In vitro evaluation of antimicrobial activity of the freshwater sponge Ochridaspongia rotunda (Arndt, 1937).

    PubMed

    Pejin, Boris; Talevski, Aleksandra; Ciric, Ana; Glamoclija, Jasmina; Nikolic, Milos; Talevski, Trajce; Sokovic, Marina

    2014-01-01

    The antimicrobial activity of five crude extracts (aqueous, methanol, ethyl acetate, acetone and methylene chloride) of the freshwater sponge Ochridaspongia rotunda (Arndt, 1937) was evaluated in vitro by using microdilution method against eight bacterial and eight fungal strains for the first time. The extracts were proven to be active in varying degrees against all the bacteria and fungi tested. O. rotunda methanol extract exhibited the highest antibacterial activity (minimum inhibitory concentration (MIC) 7.5-15.0 ?g/mL and minimum bactericidal concentration 15-30 ?g/mL), while its acetone extract exhibited the most promising antifungal activity (MIC 7.5-45.0 ?g/mL and minimum fungicidal concentration 15-60 ?g/mL). The extracts were more effective against the bacteria and fungi screened compared with the positive controls (streptomycin and ampicillin for bacteria and bifonazole and ketoconazole for fungi, respectively). According to the experimental data obtained, this deepwater sponge species may be considered as a gold mine of new antimicrobial substances with significant and broad-range activity. PMID:24804931

  12. Lysis of fresh human solid tumors by autologous lymphocytes activated in vitro with lectins

    SciTech Connect

    Mazumder, A.; Grimm, E.A.; Zhang, H.Z.; Rosenberg, S.A.

    1982-03-01

    Human peripheral blood lymphocytes (PBL), obtained from patients with a variety of cancers, were incubated in vitro with phytohemagglutinin, concanavalin A, and crude or lectin-free T-cell growth factors. The lectin-activated PBL of nine patients were capable of lysing fresh autologous tumor during a 4-hr 51Cr release assay. Multiple metastases from the same patient were equivalently lysed by these activated autologous PBL. No lysis of fresh PBL or lectin-induced lymphoblast cell targets was seen, although tumor, PBL, and lymphoblast cells were shown to be equally lysable using allosensitized cells. The activated cells could be expanded without loss of cytotoxicity in crude or lectin-free T-cell growth factors. The generation of cells lytic to fresh autologous tumor was dependent on the presence of adherent cells, although the lytic cell itself was not adherent. Proliferation was not involved in the induction of lytic cells since equal lysis was induced in irradiated and nonirradiated lymphocytes. Lectin was not required in the lytic assay, and the addition of alpha-methyl-D-mannoside to concanavalin A-activated lymphoid cells did not increase the lysis of fresh tumor cells. Activation by lectin for 3 days appears to be an efficient and convenient method for generating human cells lytic to fresh autologous tumor. These lytic cells may be of value for studies of the cell-mediated lysis of human tumor and possibly for tumor immunotherapy as well.

  13. In vitro antidiabetic activity of various crude extracts of Boletus variipes

    NASA Astrophysics Data System (ADS)

    Muniandy, Sutha; Fazry, Shazrul; Daud, Fauzi; Senafi, Sahidan

    2015-09-01

    Diabetes mellitus is a complex metabolic disease that progressively spread worldwide and difficult to treat due to various physical and metabolic complications. Current treatment using synthetic drugs has lead to various undesirable side effects. Here we determined the effect of Boletus variipes extracts on diabetes related enzymes. In this study, hot water, cold water and methanol extracts of B. variipes were utilized in order to assess their in vitro antidiabetic activity by measuring the effect on ?-amylase and ?-glucosidase enzyme. Hot water extract possessed the highest inhibition activity of ?-amylase and ?-glucosidase in a concentration dependent manner with the IC50 value 87 mg/mL and 89 mg/mL respectively. The methanol extract also showed inhibition activity of ?-amylase and ?-glucosidase but significantly lower than the hot water extract. Whereas cold water extract did not show any inhibition activity towards both the enzymes. Therefore, it is hypothesized that the hot water extract of Boletus variipes contains bioactive compound that can inhibit alpha-amylase and alpha-glucosidase enzyme activity.

  14. Cadmium inhibits the ovary ?-aminolevulinate dehydratase activity in vitro and ex vivo: protective role of seleno-furanoside.

    PubMed

    Vargas, Laura Musacchio; Soares, Melina Bucco; Izaguirry, Aryele Pinto; Lüdtke, Diogo Seibert; Braga, Hugo C; Savegnago, Lucielli; Wollenhaupt, Suzi; Brum, Daniela dos Santos; Leivas, Fábio Gallas; Santos, Francielli Weber

    2013-07-01

    Cadmium (Cd) toxicity is a concern to the tobacco-smoking sub-population which includes millions of people worldwide. Although this metal may cause severe damage to embryos and the reproductive organs, the precise mechanisms underlying its toxicity remain unclear. In the present study, the Cd effect on ovary ?-aminolevulinate dehydratase (?-ALA-D) activity was investigated in vitro and ex vivo. We observed that low concentrations of Cd inhibited cow ovary ?-ALA-D activity in vitro and the IC50 value obtained was 19.17 ?M. Furthermore, the protective effect of a novel organic selenium compound (seleno-furanoside) in restoring enzyme activity was evaluated. Seleno-furanoside (10, 50, 100, 200, 400 and 1000 ?M) did not reverse the Cd toxicity in bovine ovarian tissue in vitro. According to the in vitro reults, acute Cd exposure (2.5 and 5 mg kg(-1)) caused a significant inhibition in ovary ?-ALA-D activity in mice (around 27% and 34%, respectively). Therapy with seleno-furanoside (100 µmol kg(-1)) was able to restore enzyme activity. Thus, we demonstrated for the first time that ?-ALA-D activity from ovary is inhibited by Cd both in vitro and ex vivo. Additionally, seleno-furanoside therapy was effective in restoring ovarian enzyme activity inhibited by Cd exposure in mice, but it did not reverse the in vitro metal effect. This study detected a new toxicity marker of Cd toxicity on ovarian tissue as well as the beneficial effect of a new compound to manage the metal effect after acute exposure. PMID:22760988

  15. Phosphorylated H2AX in parthenogenetically activated, in vitro fertilized and cloned bovine embryos.

    PubMed

    Pereira, A F; Melo, L M; Freitas, V J F; Salamone, D F

    2015-08-01

    In vitro embryo production methods induce DNA damage in the embryos. In response to these injuries, histone H2AX is phosphorylated (?H2AX) and forms foci at the sites of DNA breaks to recruit repair proteins. In this work, we quantified the DNA damage in bovine embryos undergoing parthenogenetic activation (PA), in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT) by measuring ?H2AX accumulation at different developmental stages: 1-cell, 2-cell and blastocyst. At the 1-cell stage, IVF embryos exhibited a greater number of ?H2AX foci (606.1 ± 103.2) and greater area of ?H2AX staining (12923.6 ± 3214.1) than did PA and SCNT embryos. No differences at the 2-cell stage were observed among embryo types. Although PA, IVF and SCNT were associated with different blastocyst formation rates (31.1%, 19.7% and 8.3%, P < 0.05), no differences in the number of ?H2AX foci or area were detected among the treatments. ?H2AX is detected in bovine preimplantation embryos produced by PA, IVF and SCNT; the amount of DNA damage was comparable among those embryos developing to the blastocyst stage among different methods for in vitro embryo production. While IVF resulted in increased damage at the 1-cell embryo stage, no difference was observed between PA and SCNT embryos at any developmental stage. The decrease in the number of double-stranded breaks at the blastocyst stage seems to indicate that DNA repair mechanisms are functional during embryo development. PMID:24735637

  16. Microbial metabolites of omeprazole activate murine aryl hydrocarbon receptor in vitro and in vivo.

    PubMed

    Shiizaki, Kazuhiro; Kawanishi, Masanobu; Yagi, Takashi

    2014-10-01

    Omeprazole (OME), a proton pump inhibitor used to treat gastritis, is also an aryl hydrocarbon receptor (AhR) activator. OME activates AhR in human hepatocytes and hepatoma cells, but not in mice in vivo or in vitro. We recently discovered that this species-specific difference results from a difference in a few amino acids in the ligand-binding domain of AhR. However, OME activates both mouse and human AhRs in the yeast reporter assay system. Nevertheless, the cause of this discrepancy in OME responses remains unknown. Here, we report that CYP1A1 mRNA expression in mouse cecum was elevated after OME administration, although the mouse is regarded as an OME-unresponsive animal. Using the yeast reporter assay system with human and murine AhRs, we found AhR agonist-like activity in the cecal extracts of OME-treated mice. We speculated that OME metabolites produced by cecal bacteria might activate murine AhRs in vivo. In high-performance liquid chromatography (HPLC) analysis, AhR agonist-like activity of cecal bacterial culture and cecal extracts were detected at the same retention time. AhR agonist-like activity was also detected in the HPLC fractions of yeast culture media containing OME. This unknown substance could induce reporter gene expression via mouse and human AhRs. The agonist-like activity of the OME metabolite was reduced by concomitant ?-naphthoflavone exposure. These results indicate that a yeast-generated OME metabolite elicited the response of mouse AhR to OME in the yeast system, and that bacterial OME metabolites may act as AhR ligands in human and mouse intestines. PMID:25061160

  17. In vitro anti-Mycobacterium avium activities of quinolones: predicted active structures and mechanistic considerations.

    PubMed Central

    Klopman, G; Li, J Y; Wang, S; Pearson, A J; Chang, K; Jacobs, M R; Bajaksouzian, S; Ellner, J J

    1994-01-01

    The relationship between the structures of quinolones and their anti-Mycobacterium avium activities has been previously derived by using the Multiple Computer-Automated Structure Evaluation program. A number of substructural constraints required to overcome the resistance of most of the strains have been identified. Nineteen new quinolones which qualify under these substructural requirements were identified by the program and subsequently tested. The results show that the substructural attributes identified by the program produced a successful a priori prediction of the anti-M. avium activities of the new quinolones. All 19 quinolones were found to be active, and 4 of them are as active or better than ciprofloxacin. With these new quinolones, the updated multiple computer-automated structure evaluation program structure-activity relationship analysis has helped to uncover additional information about the nature of the substituents at the C5 and C7 positions needed for optimal inhibitory activity. A possible explanation of drug resistance based on the observation of suicide inactivation of bacterial cytochrome P-450 by the cyclopropylamine moiety has also been proposed and is discussed in this report. Furthermore, we confirm the view that the amount of the uncharged form present in a neutral pH solution plays a crucial role in the drug's penetration ability. PMID:7986010

  18. In Vitro Culture of Functionally Active Buffalo Hepatocytes Isolated by Using a Simplified Manual Perfusion Method

    PubMed Central

    Panda, Santanu; Bisht, Sonu; Malakar, Dhruba; Mohanty, Ashok K.; Kaushik, Jai K.

    2015-01-01

    Background In farm animals, there is no suitable cell line available to understand liver-specific functions. This has limited our understanding of liver function and metabolism in farm animals. Culturing and maintenance of functionally active hepatocytes is difficult, since they survive no more than few days. Establishing primary culture of hepatocytes can help in studying cellular metabolism, drug toxicity, hepatocyte specific gene function and regulation. Here we provide a simple in vitro method for isolation and short-term culture of functionally active buffalo hepatocytes. Results Buffalo hepatocytes were isolated from caudate lobes by using manual enzymatic perfusion and mechanical disruption of liver tissue. Hepatocyte yield was (5.3±0.66)×107 cells per gram of liver tissue with a viability of 82.3±3.5%. Freshly isolated hepatocytes were spherical with well contrasted border. After 24 hours of seeding onto fibroblast feeder layer and different extracellular matrices like dry collagen, matrigel and sandwich collagen coated plates, hepatocytes formed confluent monolayer with frequent clusters. Cultured hepatocytes exhibited typical cuboidal and polygonal shape with restored cellular polarity. Cells expressed hepatocyte-specific marker genes or proteins like albumin, hepatocyte nuclear factor 4?, glucose-6-phosphatase, tyrosine aminotransferase, cytochromes, cytokeratin and ?1-antitrypsin. Hepatocytes could be immunostained with anti-cytokeratins, anti-albumin and anti ?1-antitrypsin antibodies. Abundant lipid droplets were detected in the cytosol of hepatocytes using oil red stain. In vitro cultured hepatocytes could be grown for five days and maintained for up to nine days on buffalo skin fibroblast feeder layer. Cultured hepatocytes were viable for functional studies. Conclusion We developed a convenient and cost effective technique for hepatocytes isolation for short-term culture that exhibited morphological and functional characteristics of active hepatocytes for studying gene expression, regulation, hepatic genomics and proteomics in farm animals. PMID:25790478

  19. Voriconazole Enhances the Osteogenic Activity of Human Osteoblasts In Vitro through a Fluoride-Independent Mechanism.

    PubMed

    Allen, Kahtonna C; Sanchez, Carlos J; Niece, Krista L; Wenke, Joseph C; Akers, Kevin S

    2015-12-01

    Periostitis, which is characterized by bony pain and diffuse periosteal ossification, has been increasingly reported with prolonged clinical use of voriconazole. While resolution of clinical symptoms following discontinuation of therapy suggests a causative role for voriconazole, the biological mechanisms contributing to voriconazole-induced periostitis are unknown. To elucidate potential mechanisms, we exposed human osteoblasts in vitro to voriconazole or fluconazole at 15 or 200 ?g/ml (reflecting systemic or local administration, respectively), under nonosteogenic or osteogenic conditions, for 1, 3, or 7 days and evaluated the effects on cell proliferation (reflected by total cellular DNA) and osteogenic differentiation (reflected by alkaline phosphatase activity, calcium accumulation, and expression of genes involved in osteogenic differentiation). Release of free fluoride, vascular endothelial growth factor (VEGF), and platelet-derived growth factor (PDGF) was also measured in cell supernatants of osteoblasts exposed to triazoles, with an ion-selective electrode (for free fluoride) and enzyme-linked immunosorbent assays (ELISAs) (for VEGF and PDGF). Voriconazole but not fluconazole significantly enhanced the proliferation and differentiation of osteoblasts. In contrast to clinical observations, no increases in free fluoride levels were detected following exposure to either voriconazole or fluconazole; however, significant increases in the expression of VEGF and PDGF by osteoblasts were observed following exposure to voriconazole. Our results demonstrate that voriconazole can induce osteoblast proliferation and enhance osteogenic activity in vitro. Importantly, and in contrast to the previously proposed mechanism of fluoride-stimulated osteogenesis, our findings suggest that voriconazole-induced periostitis may also occur through fluoride-independent mechanisms that enhance the expression of cytokines that can augment osteoblastic activity. PMID:26324277

  20. Virtual Screening Analysis and In-vitro Xanthine Oxidase Inhibitory Activity of Some Commercially Available Flavonoids

    PubMed Central

    Umamaheswari, Muthuswamy; Madeswaran, Arumugam; Asokkumar, Kuppusamy

    2013-01-01

    Allopurinol, the xanthine oxidase inhibitor, is the only drug available for the treatment of gout. We examined the xanthine oxidase inhibitory activity of some commercially available flavonoids such asepigallocatechin, acacatechin, myricetin, naringenin, daidzein and glycitein by virtual screening and in-vitro studies. The interacting residues within the complex model and their contact types were identified. The virtual screening analysis were carried out using AutoDock 4.2 and in-vitro xanthine oxidase inhibitory activity was carried out using xanthine as the substrate. In addition, enzyme kinetics was performed using LineweaverBurkplot analysis. Allopurinol, a known xanthine oxidase inhibitor was used as the standard. The docking energy ofglycitein was found to be -8.49 kcal/mol which was less than that of the standard (-4.47 kcal/ mol). All the selected flavonoids were found to exhibit lower binding energy (-8.08 to -6.03 kcal/ mol) than allopurinol. The docking results confirm that flavonoids showed greater inhibition of xanthine oxidase due to their active binding sites and lesser binding energies compared to allopurinol. This may be attributed to the presence of benzopyran ring in the flavonoids. In the xanthine oxidase assay, IC50 value of glycitein was found to be 12±0.86 ?g/mL, whereas that of allopurinol was 24±0.28 ?g/mL. All the remaining compounds exhibited IC50 values ranging between 22±0.64 to 62±1.18 ?g/mL. In the enzyme kinetic studies, flavonoids showed competitive type of enzyme inhibition. It can be concluded that flavonoids could be a promising remedy for the treatment of gout and related inflammatory disorders. Further in-vivo studies are required to develop potential compounds with lesser side effects. PMID:24250638

  1. Multiparametric characterisation of neuronal network activity for in vitro agrochemical neurotoxicity assessment.

    PubMed

    Alloisio, Susanna; Nobile, Mario; Novellino, Antonio

    2015-05-01

    The last few decades have seen the marketing of hundreds of new pesticide products with a forecasted expansion of the global agrochemical industry. As several pesticides directly target nervous tissue as their mechanism of toxicity, alternative methods to routine in vivo animal testing, such as the Multi Electrode Array (MEAs)-based approach, have been proposed as an in vitro tool to perform sensitive, quick and low cost neuro-toxicological screening. Here, we examined the effects of a training set of eleven active substances known to have neuronal or non-neuronal targets, contained in the most commonly used agrochemicals, on the spontaneous electrical activity of cortical neuronal networks grown on MEAs. A multiparametric characterisation of neuronal network firing and bursting was performed with the aim of investigating how this can contribute to the efficient evaluation of in vitro chemical-induced neurotoxicity. The analysis of MFR, MBR, MBD, MISI_B and % Spikes_B parameters identified four different groups of chemicals: one wherein only inhibition is observed (chlorpyrifos, deltamethrin, orysastrobin, dimoxystrobin); a second one in which all parameters, except the MISI_B, are inhibited (carbaryl, quinmerac); a third in which increases at low chemical concentration are followed by decreases at high concentration, with exception of MISI_B that only decreased (fipronil); a fourth in which no effects are observed (paraquat, glyphosate, imidacloprid, mepiquat). The overall results demonstrated that the multiparametric description of the neuronal networks activity makes MEA-based screening platform an accurate and consistent tool for the evaluation of the toxic potential of chemicals. In particular, among the bursting parameters the MISI_B was the best that correlates with potency and may help to better define chemical toxicity when MFR is affected only at relatively high concentration. PMID:25845298

  2. Evaluation of drug-induced neurotoxicity based on metabolomics, proteomics and electrical activity measurements in complementary CNS in vitro models.

    PubMed

    Schultz, Luise; Zurich, Marie-Gabrielle; Culot, Maxime; da Costa, Anaelle; Landry, Christophe; Bellwon, Patricia; Kristl, Theresa; Hörmann, Katrin; Ruzek, Silke; Aiche, Stephan; Reinert, Knut; Bielow, Chris; Gosselet, Fabien; Cecchelli, Romeo; Huber, Christian G; Schroeder, Olaf H-U; Gramowski-Voss, Alexandra; Weiss, Dieter G; Bal-Price, Anna

    2015-12-25

    The present study was performed in an attempt to develop an in vitro integrated testing strategy (ITS) to evaluate drug-induced neurotoxicity. A number of endpoints were analyzed using two complementary brain cell culture models and an in vitro blood-brain barrier (BBB) model after single and repeated exposure treatments with selected drugs that covered the major biological, pharmacological and neuro-toxicological responses. Furthermore, four drugs (diazepam, cyclosporine A, chlorpromazine and amiodarone) were tested more in depth as representatives of different classes of neurotoxicants, inducing toxicity through different pathways of toxicity. The developed in vitro BBB model allowed detection of toxic effects at the level of BBB and evaluation of drug transport through the barrier for predicting free brain concentrations of the studied drugs. The measurement of neuronal electrical activity was found to be a sensitive tool to predict the neuroactivity and neurotoxicity of drugs after acute exposure. The histotypic 3D re-aggregating brain cell cultures, containing all brain cell types, were found to be well suited for OMICs analyses after both acute and long term treatment. The obtained data suggest that an in vitro ITS based on the information obtained from BBB studies and combined with metabolomics, proteomics and neuronal electrical activity measurements performed in stable in vitro neuronal cell culture systems, has high potential to improve current in vitro drug-induced neurotoxicity evaluation. PMID:26026931

  3. Addition of glutamate to serum free culture promotes recovery of electrical activity in adult hippocampal neurons in vitro

    PubMed Central

    Edwards, Darin; Das, Mainak; Molnar, Peter; Hickman, James J.

    2010-01-01

    A long-term cell culture system utilizing normal adult hippocampal neurons would represent an important tool that could be useful in research on the mature brain, neurological disorders and age-related neurological diseases. Historically, in vitro neuronal systems are derived from embryonic rather than mature brain tissue, a practice predicated upon difficulties in supporting regeneration, functional recovery and long-term survival of adult neurons in vitro. A few studies have shown that neurons derived from the hippocampal tissue of adult rats can survive and regenerate in vitro under serum-free conditions. However, while the adult neurons regenerated morphologically under these conditions, both the electrical activity characteristic of in vivo neurons as well as long-term neuronal survival was not consistently recovered in vitro. In this study, we report on the development of a defined culture system with the ability to support functional recovery and long-term survival of adult rat hippocampal neurons. In this system, the cell-adhesive substrate, N-1 [3-(trimethoxysilyl) propyl]-diethylenetriamine, supported neuronal attachment, regeneration, and long-term survival of adult neurons for more than 80 days in vitro. Additionally, the excitatory neurotransmitter glutamate, applied at 25 ?M for 1 to 7 days after morphological neuronal regeneration in vitro, enabled full recovery of neuronal electrical activity. This low concentration of glutamate promoted the recovery of neuronal electrical activity but with minimal excitotoxicity. These improvements allowed electrically active adult neurons to survive in vitro for several months, providing a stable test-bed for the long-term study of regeneration in adult derived neuronal systems, especially for traumatic brain injury (TBI). PMID:20452373

  4. Contact Lenses Wettability In Vitro: Effect of Surface-Active Ingredients

    PubMed Central

    Lin, Meng C.; Svitova, Tatyana F.

    2010-01-01

    Purpose To investigate the release of surface-active agents (surfactants) from unworn soft contact lenses and their influence on the lens surface wettability in vitro. Methods Surface tension (ST) of blister pack solutions was measured by pendant-drop technique. STs at the air-aqueous interface and contact angles (CAs) of four conventional and seven silicone hydrogel (SiH) soft contact lenses (SCLs) were evaluated in a dynamic-cycling regime using a modified captive-bubble tensiometer-goniometer. Measurements were performed immediately after removal from blister packs, and after soaking in a glass vial filled with a surfactant-free solution, which was replaced daily for one week. Lens surface wettability was expressed as adhesion energy (AE) according to Young’s equation. Results STs of all blister pack solutions were lower than the reference ST of pure water (72.5 mN/m), indicating the presence of surfactants. When lenses were depleted of surfactants by soaking, the STs of all studied lenses and advancing CAs of selected lenses increased (p < 0.001). Receding CAs of all studied lenses were 12° ± 5° and were not affected by the presence of surfactants. For most of the conventional lenses, the surface wettability was largely dependent on surfactants, and reduced significantly after surfactant depletion. In contrast, most SiH lenses exhibited stable and self-sustained surface wettability in vitro. Conclusions The manufacturer-added surfactants affected wetting properties of all studied SCLs, although to different degrees. PMID:20400924

  5. In Vitro Nematicidal Activity of Aryl Hydrazones and Comparative GC-MS Metabolomics Analysis.

    PubMed

    Eloh, Kodjo; Demurtas, Monica; Deplano, Alessandro; Ngoutane Mfopa, Alvine; Murgia, Antonio; Maxia, Andrea; Onnis, Valentina; Caboni, Pierluigi

    2015-11-18

    A series of aryl hydrazones were synthesized and in vitro assayed for their activity on the root-knot nematode Meloidogyne incognita. The phenylhydrazones of thiophene-2-carboxyaldehyde 5, 3-methyl-2-thiophenecarboxyaldehyde, 6, and salicylaldehyde, 2, were the most potent with EC50/48h values of 16.6 ± 2.2, 23.2 ± 2.7, and 24.3 ± 1.4 mg/L, respectively. A GC-MS metabolomics analysis, after in vitro nematode treatment with hydrazone 6 at 100 mg/L for 12 h, revealed elevated levels of fatty acids such as lauric acid, stearic acid, 2-octenoic acid, and palmitic acid. Whereas control samples showed the highest levels of monoacylglycerols such as monostearin and 2-monostearin, surprisingly, 2 h after treatment with hydrazone 6, nematodes excreted 3 times the levels of ammonia eliminated in the same conditions by controls. Thus, phenylhydrazones may represent a good scaffold in the discovery and synthesis of new nematicidal compounds, and a metabolomics approach may be helpful in understanding their mechanisms of toxicity and mode of action. PMID:26528945

  6. Synthesis and Antifungal Activity In Vitro of Isoniazid Derivatives against Histoplasma capsulatum var. capsulatum

    PubMed Central

    de Farias Marques, Francisca Jakelyne; de Aguiar Cordeiro, Rebecca; da Silva, Marcos Reinaldo; Donato Maia Malaquias, Angela; Silva de Melo, Charlline Vládia; Mafezoli, Jair; Ferreira de Oliveira, Maria da Conceiçăo; Nogueira Brilhante, Raimunda Sâmia; Gadelha Rocha, Marcos Fábio; Pinheiro Gomes Bandeira, Tereza de Jesus; Costa Sidrim, José Júlio

    2014-01-01

    Histoplasmosis is a severe infection that affects millions of patients worldwide and is endemic in the Americas. Amphotericin B (AMB) and itraconazole are highly effective for the treatment of severe and milder forms of the disease, but AMB is toxic, and the bioavailability of itraconazole is erratic. Therefore, it is important to investigate new classes of drugs for histoplasmosis treatment. In this study, a series of nine isoniazid hydrazone derivatives were synthesized and evaluated for their antifungal activities in vitro against the dimorphic fungus Histoplasma capsulatum var. capsulatum. The drugs were tested by microdilution in accordance with CLSI guidelines. The compound N?-(1-phenylethylidene)isonicotinohydrazide had the lowest MIC range of all the compounds for the yeast and filamentous forms of H. capsulatum. The in vitro synergy of this compound with AMB against the planktonic and biofilm forms of H. capsulatum cells was assessed by the checkerboard method. The effects of this hydrazone on cellular ergosterol content and membrane integrity were also investigated. The study showed that the compound alone is able to reduce the ergosterol content of planktonic cells and can alter the membrane permeability of the fungus. Furthermore, the compound alone or in combination with AMB showed inhibitory effects against mature biofilms of H. capsulatum. N?-(1-Phenylethylidene)isonicotinohydrazide alone or combined with AMB might be of interest in the management of histoplasmosis. PMID:24514090

  7. In vitro antiviral activity of Ficus carica latex against caprine herpesvirus-1.

    PubMed

    Camero, Michele; Marinaro, Mariarosaria; Lovero, Angela; Elia, Gabriella; Losurdo, Michele; Buonavoglia, Canio; Tempesta, Maria

    2014-01-01

    The latex of Ficus carica Linn. (Moraceae) has been shown to possess antiviral properties against some human viruses. To determine the ability of F. carica latex (F-latex) to interfere with the infection of caprine herpesvirus-1 (CpHV-1) in vitro, F-latex was resuspended in culture media containing 1% ethanol and was tested for potential antiviral effects against CpHV-1. Titration of CpHV-1 in the presence or in the absence of F-latex was performed on monolayers of Madin Darby Bovine Kidney (MDBK) cells. Simultaneous addition of F-latex and CpHV-1 to monolayers of MDBK cells resulted in a significant reduction of CpHV-1 titres 3 days post-infection and this effect was comparable to that induced by acyclovir. The study suggests that the F-latex is able to interfere with the replication of CpHV-1 in vitro on MDBK cells and future studies will determine the mechanisms responsible for the observed antiviral activity. PMID:24853920

  8. Immunomodulatory activity of Bengkoang (Pachyrhizus erosus) fiber extract in vitro and in vivo.

    PubMed

    Kumalasari, Ika Dyah; Nishi, Kosuke; Harmayani, Eni; Raharjo, Sri; Sugahara, Takuya

    2014-01-01

    Bengkoang (Pachyrhizus erosus (L.) Urban) is one of the most popular edible root vegetables in Indonesia. Bengkoang contains fairly large amounts of carbohydrates and crude fiber. The purpose of this research is to evaluate the immunomodulatory effect of the bengkoang fiber extract (BFE) in vitro and in vivo. BFE was prepared by heating the powder of bengkoang fiber suspended in distilled water at 121 °C for 20 min. BFE facilitated IgM production by the human hybridoma cell line HB4C5 cells. In addition, production of IgM, IgG, and IgA by mouse primary splenocytes was facilitated by BFE in a dose-dependent manner. BFE also significantly facilitated production of both interleukin-5 and interleukin-10 by splenocytes. Immunoglobulin production by lymphocytes from the spleen, Peyer's patch, and mesenteric lymph node were significantly activated by oral administration of BFE to mice for 14 days. The serum immunoglobulin levels of IgG, IgM, and IgA were also significantly enhanced. Furthermore, cytokine production by lymphocytes from the spleen, Peyer's patch, and mesenteric lymph node were also facilitated by oral administration of BFE. These results suggest that BFE has positive effects on the immune system in vitro and in vivo. PMID:23361525

  9. Everolimus is an active agent in medullary thyroid cancer: a clinical and in vitro study.

    PubMed

    Faggiano, A; Ramundo, V; Dicitore, A; Castiglioni, S; Borghi, M O; Severino, R; Ferolla, P; Crinň, L; Abbruzzese, A; Sperlongano, P; Caraglia, M; Ferone, D; Hofland, L; Colao, A; Vitale, G

    2012-07-01

    Everolimus, an mTOR inhibitor, which has been demonstrated to induce anti-tumour effects in different types of neuroendocrine tumours, has never been evaluated in patients with medullary thyroid cancer (MTC). The aim of this study was to evaluate the in vitro and in vivo effects of everolimus in combination with octreotide in MTC. Two patients with progressive metastatic MTC and high calcitonin levels were treated with everolimus 5-10 mg/day. Both patients were under treatment with octreotide LAR at the study entry. An in vitro study was also performed to assess everolimus effects on MTC cell lines (TT and MZ-CRC-1 cells). A tumour response was observed in both patients. Serum calcitonin decreased by 86% in patient 1 and by 42% in patient 2. In TT and MZ-CRC-1 cells, everolimus induced a significant dose-dependent inhibition in cell proliferation. This effect seems to be related to a cell cycle arrest in G(0) /G(1) phase in both cell lines and to the induction of cellular senescence in TT cells. Everolimus in combination with octreotide may be active as anti-tumour therapy in patients with progressive metastatic MTC, suggesting to further evaluate this agent in MTC patients in a large prospective study. PMID:21883896

  10. Everolimus is an active agent in medullary thyroid cancer: a clinical and in vitro study

    PubMed Central

    Faggiano, A; Ramundo, V; Dicitore, A; Castiglioni, S; Borghi, M O; Severino, R; Ferolla, P; Crinň, L; Abbruzzese, A; Sperlongano, P; Caraglia, M; Ferone, D; Hofland, L; Colao, A; Vitale, G

    2012-01-01

    Abstract Everolimus, an mTOR inhibitor, which has been demonstrated to induce anti-tumour effects in different types of neuroendocrine tumours, has never been evaluated in patients with medullary thyroid cancer (MTC). The aim of this study was to evaluate the in vitro and in vivo effects of everolimus in combination with octreotide in MTC. Two patients with progressive metastatic MTC and high calcitonin levels were treated with everolimus 5–10 mg/day. Both patients were under treatment with octreotide LAR at the study entry. An in vitro study was also performed to assess everolimus effects on MTC cell lines (TT and MZ-CRC-1 cells). A tumour response was observed in both patients. Serum calcitonin decreased by 86% in patient 1 and by 42% in patient 2. In TT and MZ-CRC-1 cells, everolimus induced a significant dose-dependent inhibition in cell proliferation. This effect seems to be related to a cell cycle arrest in G0/G1 phase in both cell lines and to the induction of cellular senescence in TT cells. Everolimus in combination with octreotide may be active as anti-tumour therapy in patients with progressive metastatic MTC, suggesting to further evaluate this agent in MTC patients in a large prospective study. PMID:21883896

  11. In vitro and in vivo antiplasmodial activity of three Rwandan medicinal plants and identification of their active compounds.

    PubMed

    Muganga, Raymond; Angenot, Luc; Tits, Monique; Frédérich, Michel

    2014-04-01

    In our previous study, we reported the interesting in vitro antiplasmodial activity of some Rwandan plant extracts. This gave rise to the need for these extracts to also be evaluated in vivo and to identify the compounds responsible for their antiplasmodial activity. The aim of our study was, on the one hand, to evaluate the antiplasmodial activity in vivo and the safety of the selected Rwandan medicinal plants used in the treatment of malaria, with the objective of promoting the development of improved traditional medicines and, on the other hand, to identify the active ingredients in the plants. Plant extracts were selected according to their selectivity index. The in vivo antiplasmodial activity of aqueous, methanolic, and dichloromethane extracts was then evaluated using the classical 4-day suppressive test on Plasmodium berghei infected mice. The activity of the plant extracts was estimated by measuring the percentage of parasitemia reduction, and the survival of the experimental animals was recorded. A bioguided fractionation was performed for the most promising plants, in terms of antiplasmodial activity, in order to isolate active compounds identified by means of spectroscopic and spectrometric methods. The highest level of antiplasmodial activity was observed with the methanolic extract of Fuerstia africana (>?70?%) on days 4 and 7 post-treatment after intraperitoneal injection and on day 7 using oral administration. After oral administration, the level of parasitemia reduction observed on day 4 post-infection was 44?% and 37?% with the aqueous extract of Terminalia mollis and Zanthoxylum chalybeum, respectively. However, the Z. chalybeum extract presented a high level of toxicity after intraperitoneal injection, with no animals surviving on day 1 post-treatment. F. africana, on the other hand, was safer with 40?% mouse survival on day 20 post-treatment. Ferruginol is already known as the active ingredient in F. Africana, and ellagic acid (IC50?=?175?ng/mL) and nitidine (IC50?=?77.5?ng/mL) were identified as the main active constituents of T. mollis and Z. chalybeum, respectively. F. africana presented very promising antiplasmodial activity in vivo. Although most of the plants tested showed some level of antiplasmodial activity, some of these plants may be toxic. This study revealed for the first time the role of ellagic acid and nitidine as the main antimalarial compounds in T. mollis and Z. chalybeum, respectively. PMID:24710900

  12. SJ23B, a jatrophane diterpene activates classical PKCs and displays strong activity against HIV in vitro.

    PubMed

    Bedoya, Luis M; Márquez, Nieves; Martínez, Natalia; Gutiérrez-Eisman, Silvia; Alvarez, Amparo; Calzado, Marco A; Rojas, José M; Appendino, Giovanni; Muńoz, Eduardo; Alcamí, José

    2009-03-15

    Existence of virus reservoirs makes the eradication of HIV infection extremely difficult. Current drug therapies neither eliminate these viral reservoirs nor prevent their formation. Consequently, new strategies are needed to target these reservoirs with the aim of decreasing their size. We analysed a series of jatrophane diterpenes isolated from Euphorbia hyberna and we found that one of them, SJ23B, induces the internalization of the HIV-1 receptors CD4, CXCR4 and CCR5 and prevents R5 and X4 viral infection in human primary T cells at the nanomolar range. Moreover, SJ23B is a potent antagonist of HIV-1 latency. Using Jurkat-LAT-GFP cells, a model for HIV-1 latency, we found that prostratin and SJ23B activate HIV-1 gene expression, with SJ23B being at least 10-fold more potent than prostratin. SJ23B did not elicit transforming foci activity in NIH 3T3 cells but is a potent activator of PKCalpha and delta as measured by in vitro kinase assays and by cellular translocation experiments. By using isoform-specific PKC inhibitors we found that cPKCs are critical for SJ23B-induced HIV-1 reactivation. We also showed that both SJ23B-induced IkappaBalpha degradation and NF-kappaB activation were inhibited by the classical PKC inhibitor, Gö6976. Accordingly, SJ23B synergizes with ionomycin to translocate PKCalpha to the plasma membrane and to activate the NF-kappaB pathway. Moreover, SJ23B activates both NF-kappaB and Sp1-dependent transcriptional activities in primary T cells. We have shown that diterpene jatrophanes represent a new member of anti-AIDS agents that could be developed for mitigating HIV reactivation. PMID:19100719

  13. In vitro evaluation of Portuguese propolis and floral sources for antiprotozoal, antibacterial and antifungal activity.

    PubMed

    Falcăo, Soraia I; Vale, Nuno; Cos, Paul; Gomes, Paula; Freire, Cristina; Maes, Louis; Vilas-Boas, Miguel

    2014-03-01

    Propolis is a beehive product with a very complex chemical composition, used since ancient times in several therapeutic treatments. As a contribution to the improvement of drugs against several tropical diseases caused by protozoa, we screened Portuguese propolis and its potential floral sources Populus x Canadensis and Cistus ladanifer against Plasmodium falciparum, Leishmania infantum, Trypanosoma brucei and Trypanosoma cruzi. The toxicity against MRC-5 fibroblast cells was evaluated to assess selectivity. The in vitro assays were performed following the recommendations of WHO Special Programme for Research and Training in Tropical Diseases (TDR) and revealed moderate activity, with the propolis extracts presenting the relatively highest inhibitory effect against T. brucei. Additionally, the antimicrobial activity against Staphylococcus aureus, Candida albicans, Trichophyton rubrum and Aspergillus fumigatus was also verified with the better results observed against T. rubrum. The quality of the extracts was controlled by evaluating the phenolic content and antioxidant activity. The observed biological activity variations are associated with the variable chemical composition of the propolis and the potential floral sources under study. PMID:23722631

  14. Influence of cryopreservation on the antioxidative activity of in vitro cultivated Hypericum species

    PubMed Central

    Georgieva, Elena; Petrova, Detelina; Yordanova, Zhenya; Kapchina-Toteva, Veneta; Cellarova, Eva; Chaneva, Ganka

    2014-01-01

    Antioxidative activity of two in vitro cultivated Hypericum species – H. rumeliacum Boiss. and H. tetrapterum Fr. – was estimated after cryopreservation. Both species were successfully regenerated after a cryopreservation procedure performed by the vitrification method. H. tetrapterum did not manifest any significant oxidative stress-induced changes caused by low-temperature treatment. Conversely, a decrease in green pigments' content of H. rumeliacum was measured, particularly pronounced in chlorophyll b, which was accompanied by an increase of carotenoids in the regenerated plants. A strong increase of malone dialdehyde and H2O2 levels in H. rumeliacum tissues was detected. Superoxide dismutase activity was enhanced by 170%, as well as the catalase activity, which was 220% above the control. The same trend was observed in H. tetrapterum, although less pronounced – 143% increase of superoxide dismutase and 112% of catalase. Cryopreservation did not influence the phenol content in the examined plants, but it led to an increase of flavonoid content, especially in H. tetrapterum, by 237%. Total antioxidant activity in regenerated H. tetrapterum varied around the control level, but it was increased in H. rumeliacum. The free proline content in H. tetrapterum remained almost unaffected after freezing, as opposed to H. rumeliacum, where a strong increase of proline content (208% above the control) occurred. An electrolyte leakage from the cells of H. rumeliacum regenerated after cryopreservation was also registered, albeit not significant.

  15. In vitro assessment of antioxidant activity of tyrosol, resveratrol and their acetylated derivatives.

    PubMed

    Vlachogianni, Ioanna C; Fragopoulou, Elizabeth; Kostakis, Ioannis K; Antonopoulou, Smaragdi

    2015-06-15

    Consumption of phenolic compounds is associated with beneficial effects in humans even though many of them are poorly absorbed. The aim of this study was to investigate the in vitro antioxidant activity of tyrosol (T), resveratrol (R) and their acetylated derivatives (AcD), as increased lipophilicity has been reported to improve absorption. The chemically synthesized AcDs were evaluated by their ability to scavenge DPPH radicals, inhibit non-enzymatic linoleic acid peroxidation, inhibit human serum oxidation in the presence of copper ions and inhibit lipoxygenase activity. T showed an inhibitory effect only in serum oxidation, where the T-acetylated at aromatic-OH was the most active. The T-acetylated at aliphatic-OH and 3,5-diacetyl-R exhibited the most powerful effect in non-enzymatic linoleic acid peroxidation with IC50 values 2.4 mM ± 0.21 and 0.055 mM ± 0.0018, respectively. In all other tests R was the most potent among all its AcD and T. Increasing lipophilicity by acetylation improves antioxidant activity of phenolic compounds in non-enzymatic lipid peroxidation assays. PMID:25660873

  16. A topological study of repetitive co-activation networks in in vitro cortical assemblies

    NASA Astrophysics Data System (ADS)

    Pirino, Virginia; Riccomagno, Eva; Martinoia, Sergio; Massobrio, Paolo

    2015-02-01

    To address the issue of extracting useful information from large data-set of large scale networks of neurons, we propose an algorithm that involves both algebraic-statistical and topological tools. We investigate the electrical behavior of in vitro cortical assemblies both during spontaneous and stimulus-evoked activity coupled to Micro-Electrode Arrays (MEAs). Our goal is to identify core sub-networks of repetitive and synchronous patterns of activity and to characterize them. The analysis is performed at different resolution levels using a clustering algorithm that reduces the network dimensionality. To better visualize the results, we provide a graphical representation of the detected sub-networks and characterize them with a topological invariant, i.e. the sequence of Betti numbers computed on the associated simplicial complexes. The results show that the extracted sub-populations of neurons have a more heterogeneous firing rate with respect to the entire network. Furthermore, the comparison of spontaneous and stimulus-evoked behavior reveals similarities in the identified clusters of neurons, indicating that in both conditions similar activation patterns drive the global network activity.

  17. In vitro activity of a Combretum micranthum extract against herpes simplex virus types 1 and 2.

    PubMed

    Ferrea, G; Canessa, A; Sampietro, F; Cruciani, M; Romussi, G; Bassetti, D

    1993-08-01

    The authors demonstrate in vitro antiviral activity of a methanolic extract of Combretum micranthum leaves against HSV-1 and HSV-2. This activity is present only in the extract dissolved 7 days before the assay, but not in the freshly prepared extract, thus indicating the presence of inactive precursors which undergo spontaneous transformations into active compounds. The alkaline autooxidation of the methanolic extract promotes this rapid transformation. The precursors have been identified as condensed catechinic tannins, which, under alkaline conditions, suffer rapid cleavage, intramolecular rearrangement to catechinic acid and autooxidation. The alkaline autooxidation products of the methanolic extract of C. micranthum and those of the synthetic catechinic acid show similar I.R. and U.V. absorption curves, as well as similar anti-HSV-1 and -HSV-2 activities. EC50s of catechinic acid autooxidation products against HSV-1 and HSV-2 replication were 2 micrograms/ml and 4 micrograms/ml, respectively, when cell cultures were treated with the compound during virus infection. PMID:8215303

  18. Ligand Binding and Activation of PPAR? by Firemaster® 550: Effects on Adipogenesis and Osteogenesis in Vitro

    PubMed Central

    Pillai, Hari K.; Fang, Mingliang; Beglov, Dmitri; Kozakov, Dima; Vajda, Sandor; Stapleton, Heather M.; Webster, Thomas F.

    2014-01-01

    Background: The use of alternative flame retardants has increased since the phase out of pentabromodiphenyl ethers (pentaBDEs). One alternative, Firemaster® 550 (FM550), induces obesity in rats. Triphenyl phosphate (TPP), a component of FM550, has a structure similar to that of organotins, which are obesogenic in rodents. Objectives: We tested the hypothesis that components of FM550 are biologically active peroxisome proliferator-activated receptor ? (PPAR?) ligands and estimated indoor exposure to TPP. Methods: FM550 and its components were assessed for ligand binding to and activation of human PPAR?. Solvent mapping was used to model TPP in the PPAR? binding site. Adipocyte and osteoblast differentiation were assessed in bone marrow multipotent mesenchymal stromal cell models. We estimated exposure of children to TPP using a screening-level indoor exposure model and house dust concentrations determined previously. Results: FM550 bound human PPAR?, and binding appeared to be driven primarily by TPP. Solvent mapping revealed that TPP interacted with binding hot spots within the PPAR? ligand binding domain. FM550 and its organophosphate components increased human PPAR?1 transcriptional activity in a Cos7 reporter assay and induced lipid accumulation and perilipin protein expression in BMS2 cells. FM550 and TPP diverted osteogenic differentiation toward adipogenesis in primary mouse bone marrow cultures. Our estimates suggest that dust ingestion is the major route of exposure of children to TPP. Conclusions: Our findings suggest that FM550 components bind and activate PPAR?. In addition, in vitro exposure initiated adipocyte differentiation and antagonized osteogenesis. TPP likely is a major contributor to these biological actions. Given that TPP is ubiquitous in house dust, further studies are warranted to investigate the health effects of FM550. Citation: Pillai HK, Fang M, Beglov D, Kozakov D, Vajda S, Stapleton HM, Webster TF, Schlezinger JJ. 2014. Ligand binding and activation of PPAR? by Firemaster® 550: effects on adipogenesis and osteogenesis in vitro. Environ Health Perspect 122:1225–1232;?http://dx.doi.org/10.1289/ehp.1408111 PMID:25062436

  19. In vitro Activation of Heat Shock Transcription Factor DNA-Binding by Calcium and Biochemical Conditions that Affect Protein Conformation

    NASA Astrophysics Data System (ADS)

    Mosser, Dick D.; Kotzbauer, Paul T.; Sarge, Kevin D.; Morimoto, Richard I.

    1990-05-01

    The transcription of heat shock genes in response to physiological stress requires activation of heat shock transcription factor (HSF). Although the transcriptional response is most commonly induced by temperature elevation, the biochemical events involved in HSF activation in vivo can also be triggered at normal physiological temperatures by chemicals that inhibit metabolic processes. We have used a HeLa cell-free system in which HSF DNA-binding is activated by conditions that affect protein conformation, including increasing concentrations of hydrogen ions, urea, or nonionic detergents. Treatment with calcium ions also results in a concentration- and time-dependent activation of HSF in vitro. Pretreatment with each of these biochemical conditions reduces the temperature dependence for HSF activation in vitro. These results suggest that HSF is activated either directly by under-going a conformational change or indirectly through interactions with unfolded proteins.

  20. Systemic immune responses in Alzheimer's disease: in vitro mononuclear cell activation and cytokine production.

    PubMed

    Pellicanň, Mariavaleria; Bulati, Matteo; Buffa, Silvio; Barbagallo, Mario; Di Prima, Anna; Misiano, Gabriella; Picone, Pasquale; Di Carlo, Marta; Nuzzo, Domenico; Candore, Giuseppina; Vasto, Sonya; Lio, Domenico; Caruso, Calogero; Colonna-Romano, Giuseppina

    2010-01-01

    To investigate the systemic signs of immune-inflammatory responses in Alzheimer's disease (AD), in the present study we have analyzed blood lymphocyte subsets and the expression of activation markers on peripheral blood mononuclear cells (PBMCs) from AD patients and age-matched healthy controls (HC) activated in vitro by recombinant amyloid-beta peptide (rAbeta42). Our study of AD lymphocyte subpopulations confirms the already described decrease of the absolute number and percentage of B cells when compared to HC lymphocytes, whereas the other subsets are not significantly different in patients and controls. We report the increased expression of the activation marker CD69 and of the chemokine receptors CCR2 and CCR5 on T cells but no changes of CD25 after activation. B cells are also activated by rAbeta42 as demonstrated by the enhanced expression of CCR5. Moreover, rAbeta42 induces an increased expression of the scavenger receptor CD36 on monocytes. Some activation markers and chemokine receptors are overexpressed in unstimulated AD cells when compared to controls. This is evidence of the pro-inflammatory status of AD. Stimulation by rAbeta42 also induces the production of the pro-inflammatory cytokines IL-1beta, IL-6, IFN-gamma, and TNF-alpha, and of the anti-inflammatory cytokines IL-10 and IL-1Ra. The chemokines RANTES, MIP-1beta, and eotaxin as well as some growth factors (GM-CSF, G-CSF) are also overproduced by AD-derived PBMC activated by rAbeta42. These results support the involvement of systemic immunity in AD patients. However, our study is an observational one so we cannot draw a conclusion about its contribution to the pathophysiology of the disease. PMID:20413861

  1. Activation of Neuropeptide Y Receptors Modulates Retinal Ganglion Cell Physiology and Exerts Neuroprotective Actions In Vitro

    PubMed Central

    Martins, Joăo; Elvas, Filipe; Brudzewsky, Dan; Martins, Tânia; Kolomiets, Bogdan; Tralhăo, Pedro; Gřtzsche, Casper R.; Cavadas, Cláudia; Castelo-Branco, Miguel; Woldbye, David P. D.; Picaud, Serge; Santiago, Ana R.

    2015-01-01

    Neuropeptide Y (NPY) is expressed in mammalian retina but the location and potential modulatory effects of NPY receptor activation remain largely unknown. Retinal ganglion cell (RGC) death is a hallmark of several retinal degenerative diseases, particularly glaucoma. Using purified RGCs and ex vivo rat retinal preparations, we have measured RGC intracellular free calcium concentration ([Ca2+]i) and RGC spiking activity, respectively. We found that NPY attenuated the increase in the [Ca2+]i triggered by glutamate mainly via Y1 receptor activation. Moreover, (Leu31, Pro34)?NPY, a Y1/Y5 receptor agonist, increased the initial burst response of OFF-type RGCs, although no effect was observed on RGC spontaneous spiking activity. The Y1 receptor activation was also able to directly modulate RGC responses by attenuating the NMDA-induced increase in RGC spiking activity. These results suggest that Y1 receptor activation, at the level of inner or outer plexiform layers, leads to modulation of RGC receptive field properties. Using in vitro cultures of rat retinal explants exposed to NMDA, we found that NPY pretreatment prevented NMDA-induced cell death. However, in an animal model of retinal ischemia-reperfusion injury, pretreatment with NPY or (Leu31, Pro34)?NPY was not able to prevent apoptosis or rescue RGCs. In conclusion, we found modulatory effects of NPY application that for the first time were detected at the level of RGCs. However, further studies are needed to evaluate whether NPY neuroprotective actions detected in retinal explants can be translated into animal models of retinal degenerative diseases. PMID:26311075

  2. In vitro antagonistic activity of monoterpenes and their mixtures against 'toe nail fungus' pathogens.

    PubMed

    Ramsewak, Russel S; Nair, Muraleedharan G; Stommel, Manfred; Selanders, Louise

    2003-04-01

    The antibiotic effect of the active ingredients in Meijer medicated chest rub (eucalyptus oil, camphor and menthol) as well as the inactive ingredients (thymol, oil of turpentine, oil of nutmeg and oil of cedar leaf) were studied in vitro using the fungal pathogens responsible for onychomycosis, such as the dermatophytes Tricophyton rubrum, Trichophyton mentagrophytes, Microsporum canis, Epidermophyton fl occosum and Epidermophyton stockdale. The zones of inhibition data revealed that camphor (1). menthol (2). thymol (3). and oil of Eucalyptus citriodora were the most efficacious components against the test organisms. The MIC(100) for mixtures of these four components in various carrier solvents revealed that formulations consisting of 5 mg/mL concentrations of each have a potential to be efffective in controlling onychomycosis. PMID:12722144

  3. In vitro activity of carvacrol and thymol combined with antifungals or antibacterials against Pythium insidiosum.

    PubMed

    Jesus, F P K; Ferreiro, L; Bizzi, K S; Loreto, É S; Pilotto, M B; Ludwig, A; Alves, S H; Zanette, R A; Santurio, J M

    2015-06-01

    We describe the in vitro activities of the combinations of carvacrol and thymol with antibiotics (azithromycin, clarithromycin, minocycline and tigecycline) and antifungal agents (amphotericin B, caspofungin, itraconazole and terbinafine) against 23 isolates of the oomycete Pythium insidiosum. The assays were based on the M38-A2 technique and checkerboard microdilution. Based on the mean FICI values, the main synergies observed were combinations of carvacrol+itraconazole and thymol+itraconazole (96%), thymol+clarithromycin (92%), carvacrol+clarithromycin (88%), thymol+minocycline (84%), carvacrol+minocycline (80%), carvacrol+azithromycin (76%), thymol+azithromycin (68%), carvacrol+tigecycline (64%) and thymol+tigecycline (60%). In conclusion, we found that combinations of carvacrol or thymol with these antimicrobial agents might provide effective alternative treatments for cutaneous pythiosis due to their synergistic interactions. Future in vivo experiments are needed to elucidate the safety and therapeutic potential of these combinations. PMID:25639921

  4. Synthesis, characterization, DNA interaction and in vitro cytotoxicity activities of ruthenium(II) Schiff base complexes

    NASA Astrophysics Data System (ADS)

    Sathiyaraj, Subbaiyan; Butcher, Ray J.; Jayabalakrishnan, Chinnasamy

    2012-12-01

    DNA binding, cleavage and cytotoxicity characteristics of a novel Schiff base ligand 3-(benzothiazol-2-yliminomethyl)-naphthalen-2-ol and ruthenium(II) complexes have been investigated. The DNA interaction properties of the complexes have been investigated using absorption spectra, as well as gel electrophoresis studies. Intrinsic binding constant (Kb) has been estimated under similar set of experimental conditions. Absorption spectral study indicate that the ligand and ruthenium(II) complexes has intrinsic binding constant in the range of 1.4-7.2 × 104 M-1. Ruthenium(II) complexes show more binding ability than the ligand. Further, in vitro cytotoxicity study of the ligand and the complexes exhibited antitumor activity against HeLa and HEp2 tumor cells.

  5. In Vitro Activities of Nine Antifungal Drugs and Their Combinations against Phialophora verrucosa

    PubMed Central

    Li, Yali; Wan, Zhe

    2014-01-01

    The in vitro activities of nine antifungal drugs and their combinations against 31 clinical and 15 environmental Phialophora verrucosa strains were tested. The MIC90/90% minimum effective concentration (MIC/MEC90) values (?g/ml) across all strains were as follows: for terbinafine, 0.25; for posaconazole, 0.5; for voriconazole, 1; for itraconazole, 2; for amphotericin B, 4; for caspofungin and micafungin, 16; and for fluconazole and flucytosine, 64. The highest synergy was shown by the combination of itraconazole plus caspofungin (with synergy against 100% of the 31 clinical strains), followed by amphotericin B plus flucytosine (45.2%) and itraconazole plus terbinafine or micafungin (25.8% or 12.9%, respectively). PMID:24982078

  6. In vitro antimicrobial activity of Romanian medicinal plants hydroalcoholic extracts on planktonic and adhered cells.

    PubMed

    Stanciuc, A M; Gaspar, A; Moldovan, L; Saviuc, C; Popa, M; M?ru?escu, L

    2011-01-01

    The aim of this study was to assess the antibacterial and antifungal potential of some Romanian medicinal plants, arnica--Arnica montana, wormwood--Artemisia absinthium and nettle--Urtica dioica. In order to perform this antimicrobial screening, we obtained the vegetal extracts and we tested them on a series of Gram-positive and Gram-negative bacteria, and also against two fungal strains. The vegetal extracts showed antimicrobial activity preferentially directed against the planktonic fungal and bacterial growth, while the effect against biofilm formation and development was demonstrated only against S. aureus and C. albicans. Our in vitro assays indicate that the studied plant extracts are a significant source of natural alternatives to antimicrobial therapy, thus avoiding antibiotic therapy, the use of which has become excessive in recent years. PMID:21717806

  7. Comparison of essential oil components and in vitro anticancer activity in wild and cultivated Salvia verbenaca.

    PubMed

    Russo, Alessandra; Cardile, Venera; Graziano, Adriana C E; Formisano, Carmen; Rigano, Daniela; Canzoneri, Marisa; Bruno, Maurizio; Senatore, Felice

    2015-01-01

    The objectives of our research were to study the chemical composition and the in vitro anticancer effect of the essential oil of Salvia verbenaca growing in natural sites in comparison with those of cultivated (Sc) plants. The oil from wild (Sw) S. verbenaca presented hexadecanoic acid (23.1%) as the main constituent, while the oil from Sc plants contained high quantities of hexahydrofarnesyl acetone (9.7%), scarce in the natural oil (0.7%). The growth-inhibitory and proapoptotic effects of the essential oils from Sw and Sc S. verbenaca were evaluated in the human melanoma cell line M14, testing cell vitality, cell membrane integrity, genomic DNA fragmentation and caspase-3 activity. Both the essential oils were able to inhibit the growth of the cancer cells examined inducing also apoptotic cell death, but the essential oil from cultivated samples exhibited the major effects. PMID:25537231

  8. In Vitro Activity of Nemonoxacin, a Novel Nonfluorinated Quinolone Antibiotic, against Chlamydia trachomatis and Chlamydia pneumoniae

    PubMed Central

    Chotikanatis, Kobkul; Kohlhoff, Stephan A.

    2014-01-01

    The in vitro activities of nemonoxacin, levofloxacin, azithromycin, and doxycycline were tested against 10 isolates each of Chlamydia trachomatis and Chlamydia pneumoniae. The MICs at which 90% of the isolates of both C. trachomatis and C. pneumoniae were inhibited (MIC90s) were 0.06 ?g/ml (range, 0.03 to 0.13 ?g/ml). The minimal bactericidal concentrations at which 90% of the isolates were killed by nemonoxacin (MBC90s) were 0.06 ?g/ml for C. trachomatis (range, 0.03 to 0.125 ?g/ml) and 0.25 for C. pneumoniae (range, 0.015 to 0.5 ?g/ml). PMID:24366753

  9. In Vitro Activity of AZD5847 against Geographically Diverse Clinical Isolates of Mycobacterium tuberculosis

    PubMed Central

    Wijkander, Maria; Perskvist, Nasrin; Balasubramanian, V.; Sambandamurthy, Vasan K.; Hoffner, Sven

    2014-01-01

    The MIC of the novel antituberculosis (anti-TB) drug AZD5847 was determined against 146 clinical isolates from diverse geographical regions, including eastern Europe, North America, Africa, and Asia, using the automated Bactec Mycobacterial Growth Indicator Tube (MGIT) 960 system. These isolates originated from specimen sources such as sputum, bronchial alveolar lavage fluid, pleural fluid, abscess material, lung biopsies, and feces. The overall MIC90 was 1.0 mg/liter (range, 0.125 to 4 mg/liter). The MICs of AZD5847 for isolates of Mycobacterium tuberculosis were similar among drug-sensitive strains, multidrug-resistant (MDR) strains, and extensively drug resistant (XDR) strains. The good in vitro activity of AZD5847 against M. tuberculosis and the lack of cross-resistance make this agent a promising anti-TB drug candidate. PMID:24777103

  10. Designed, synthetically accessible bryostatin analogues potently induce activation of latent HIV reservoirs in vitro

    NASA Astrophysics Data System (ADS)

    Dechristopher, Brian A.; Loy, Brian A.; Marsden, Matthew D.; Schrier, Adam J.; Zack, Jerome A.; Wender, Paul A.

    2012-09-01

    Bryostatin is a unique lead in the development of potentially transformative therapies for cancer, Alzheimer's disease and the eradication of HIV/AIDS. However, the clinical use of bryostatin has been hampered by its limited supply, difficulties in accessing clinically relevant derivatives, and side effects. Here, we address these problems through the step-economical syntheses of seven members of a new family of designed bryostatin analogues using a highly convergent Prins-macrocyclization strategy. We also demonstrate for the first time that such analogues effectively induce latent HIV activation in vitro with potencies similar to or better than bryostatin. Significantly, these analogues are up to 1,000-fold more potent in inducing latent HIV expression than prostratin, the current clinical candidate for latent virus induction. This study provides the first demonstration that designed, synthetically accessible bryostatin analogues could serve as superior candidates for the eradication of HIV/AIDS through induction of latent viral reservoirs in conjunction with current antiretroviral therapy.

  11. In vitro anticancer activity of extracts of Mentha Spp. against human cancer cells.

    PubMed

    Sharma, Vikas; Hussain, Shabir; Gupta, Moni; Saxena, Ajit Kumar

    2014-10-01

    In vitro anticancer potential of methanolic and aqueous extracts of whole plants of Mentha arvensis, M. longifolia, M. spicata and M. viridis at concentration of 100 ?g/ml was evaluated against eight human cancer cell lines--A-549, COLO-205, HCT-116, MCF-7, NCI-H322, PC-3, THP-1 and U-87MG from six different origins (breast, colon, glioblastoma, lung, leukemia and prostate) using sulphorhodamine blue (SRB) assay. Methanolic extracts of above-mentioned Mentha Spp. displayed anti-proliferative effect in the range of 70-97% against four human cancer cell lines, namely COLO-205, MCF-7, NCI-H322 and THP-1; however, aqueous extracts were found to be active against HCT-116 and PC-3. The results indicate that Mentha Spp. contain certain constituents with cytotoxic properties which may find use in developing anticancer agents. PMID:25630112

  12. Anti-diabetic activity of a mineraloid isolate, in vitro and in genetically diabetic mice.

    PubMed

    Deneau, Joel; Ahmed, Taufeeq; Blotsky, Roger; Bojanowski, Krzysztof

    2011-01-01

    Type II diabetes is a metabolic disease mediated through multiple molecular pathways. Here, we report anti-diabetic effect of a standardized isolate from a fossil material - a mineraloid leonardite - in in vitro tests and in genetically diabetic mice. The mineraloid isolate stimulated mitochondrial metabolism in human fibroblasts and this stimulation correlated with enhanced expression of genes coding for mitochondrial proteins such as ATP synthases and ribosomal protein precursors, as measured by DNA microarrays. In the diabetic animal model, consumption of the Totala isolate resulted in decreased weight gain, blood glucose, and glycated hemoglobin. To our best knowledge, this is the first description ever of a fossil material having anti-diabetic activity in pre-clinical models. PMID:22002216

  13. Evaluation of a shock wave induced cavitation activity both in vitro and in vivo.

    PubMed

    Tu, Juan; Matula, Thomas J; Bailey, Michael R; Crum, Lawrence A

    2007-10-01

    This study evaluated the cavitation activity induced by shock wave (SW) pulses, both in vitro and in vivo, based on the area measurements of echogenic regions observed in B-mode ultrasound images. Residual cavitation bubble clouds induced by SW pulses were detected as echogenic regions in B-mode images. The temporal evolution of residual bubble clouds, generated by SWs with varying lithotripter charging voltage and pulse repetition frequency (PRF), was analyzed by measuring the time-varying behaviors of the echogenic region areas recorded in B-mode images. The results showed that (1) the area of SW-induced echogenic regions enlarged with increased SW pulse number; (2) echogenic regions in the B-mode images dissipated gradually after ceasing the SWs, which indicated the dissolution of the cavitation bubbles; and (3) larger echogenic regions were generated with higher charging voltage or PRF. PMID:17881810

  14. In vitro and in vivo anti-angiogenic activity of girinimbine isolated from Murraya koenigii

    PubMed Central

    Iman, Venoos; Karimian, Hamed; Mohan, Syam; Hobani, Yahya Hasan; Noordin, Mohamed Ibrahim; Mustafa, Mohd Rais; Noor, Suzita Mohd

    2015-01-01

    Girinimbine is a carbazole alkaloid isolated from the stem bark and root of Murraya koenigii. Here we report that girinimbine is an inhibitor of angiogenic activity both in vitro and in vivo. MTT results showed that girinimbine inhibited proliferation of human umbilical vein endothelial cells, while results from endothelial cell invasion, migration, tube formation, and wound healing assays demonstrated significant time- and dose-dependent inhibition by girinimbine. A proteome profiler array done on girinimbine-treated human umbilical vein endothelial cells showed that girinimbine had mediated regulation of pro-angiogenic and anti-angiogenic proteins. The anti-angiogenic potential of girinimbine was also evidenced in vivo in the zebrafish embryo model wherein girinimbine inhibited neo vessel formation in zebrafish embryos following 24 hours of exposure. Together, these results showed that girinimbine could effectively suppress angiogenesis, suggestive of its therapeutic potential as a novel angiogenesis inhibitor. PMID:25767375

  15. In vitro antibacterial activity of Camellia sinensis extract against cariogenic microorganisms

    PubMed Central

    Anita, P.; Sivasamy, Shyam; Madan Kumar, P. D.; Balan, I. Nanda; Ethiraj, Sumathi

    2014-01-01

    Context: Dental caries, a ubiquitous multifactorial infectious disease, is primarily caused by microorganisms like Streptococcus mutans and Lactobacillus acidophilus. Use of antimicrobials is an important strategy to curb cariogenic microorganisms. Aim: The aim was to evaluate the in vitro antimicrobial activity of C. sinensis extract on S. mutans and L. acidophilus. Study Setting and Design: Experimental design, in vitro study, lab setting. Materials and Methods: Aqueous, acetone and ethanolic extracts of C. sinensis were subjected to antioxidant analysis. The ethanolic extract was used for assessment of antimicrobial properties. Ethanolic green tea extract at ten different concentrations and 0.2% chlorhexidine was used. Microbiological investigations were carried out to determine the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and zone of Inhibition of the test and control agents against S. mutans and L. acidophilus. Statistical Analysis: Kruskall–Wallis and Mann–Whitney U-test. Results: MIC of green tea extract on S. mutans and L. acidophilus was found to be 0.2% and 0.3% respectively, MBC was found to be 0.8% and 0.9%, respectively. The mean zone of inhibition for 30 ?l containing 300 ?g of ethanolic extract of green tea and control against S. mutans were 18.33 mm and 14.67 mm, respectively. The mean zone of inhibition for 30 ?l containing 300 ?g of ethanolic extract of green tea and control against L. acidophilus were 12.67 mm and 7.33 mm, respectively. Conclusion: Green tea has antibacterial activity against predominant cariogenic bacteria namely S. mutans and L. acidophilus. PMID:25538470

  16. In vitro assay for osteoinductive activity of different demineralized freeze-dried bone allograft

    PubMed Central

    Vaziri, Shahram; Vahabi, Surena; Torshabi, Maryam

    2012-01-01

    Purpose Various bone graft materials have been used for periodontal tissue regeneration. Demineralized freeze-dried bone allograft (DFDBA) is a widely used bone substitute. The current widespread use of DFDBA is based on its potential osteoinductive ability. Due to the lack of verifiable data, the purpose of this study was to assess the osteoinductive activity of different DFDBAs in vitro. Methods Sarcoma osteogenic (SaOS-2) cells (human osteoblast-like cells) were exposed to 8 mg/mL and 16 mg/mL concentrations of three commercial types of DFDBA: Osseo+, AlloOss, and Cenobone. The effect of these materials on cell proliferation was determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The osteoinductive ability was evaluated using alizarin red staining, and the results were confirmed by evaluating osteogenic gene expression using reverse transcription polymerase chain reaction (RT-PCR). Results In the SaOS-2 cells, an 8 mg/mL concentration of Osseo+ and Cenobone significantly increased cell proliferation in 48 hours after exposure (P<0.001); however, in these two bone materials, the proliferation of cells was significantly decreased after 48 hours of exposure with a 16 mg/mL concentration (P<0.001). The alizarin red staining results demonstrated that the 16 mg/mL concentration of all three tested DFDBA induced complete morphologic differentiation and mineralized nodule production of the SaOS-2 cells. The RT-PCR results revealed osteopontin gene expression at a 16 mg/mL concentration of all three test groups, but not at an 8 mg/mL concentration. Conclusions These commercial types of DFDBA are capable of decreasing proliferation and increasing osteogenic differentiation of the SaOS-2 cell line and have osteoinductive activity in vitro. PMID:23346466

  17. In vitro anthelmintic activity of Zanthoxylum simulans essential oil against Haemonchus contortus.

    PubMed

    Qi, H; Wang, W X; Dai, J L; Zhu, L

    2015-07-30

    The need for new anthelmintic agents with low impact on the environment is becoming urgent. Phytotherapy is an alternative method to control gastrointestinal nematodes in small ruminants. This study aims to determine the composition of Zanthoxylum simulans essential oil (ZSEO) and evaluate the in vitro ovicidal and larvicidal effects of ZSEO on Haemonchus contortus using egg hatch assay, larval development assay (LDA), and larval migration inhibition assay (LMIA). The chemical composition of ZSEO was determined through gas chromatography and mass spectrometry and 94 compounds were identified from the ZSEO. The major constituents of ZSEO were borneol (18.61%), ?-elemene (10.87%). ZSEO and borneol both at 40 mg/mL inhibited larval hatching by 100%, with LC50 values of 3.98 and 1.50mg/mL, respectively. The LC50 value of ?-elemene was not determined because of its insufficient activity. The results of LDA showed that ZSEO, borneol, and ?-elemene all at 40 mg/mL inhibited larval development by 99.8%, 100%, and 55.4%, respectively, and exhibited dose-dependent responses with LC50 values of 4.02, 1.99, and 32.17 mg/mL, respectively. The results of LMIA showed that ZSEO, borneol, and ?-elemene all at 40 mg/mL inhibited larval migration by 74.3%, 97.0%, and 53.2%, respectively. ZSEO presented ovicidal and larvicidal activities in vitro. Therefore, Zanthoxylum may be an alternative source of anthelmintic agents to control gastrointestinal nematodes in sheep. PMID:26073109

  18. Studies on the in vitro and in vivo antiurolithic activity of Holarrhena antidysenterica

    PubMed Central

    Khan, Aslam; Khan, Saeed R.; Gilani, Anwar H.

    2013-01-01

    Background Holarrhena antidysenterica has a traditional use in the treatment of urolithiasis, therefore, its crude extract has been investigated for possible antiurolithic effect. Materials and methods The crude aqueous-methanolic extract of Holarrhena antidysenterica (Ha.Cr) was studied using the in vitro and in vivo methods. Results In the in vitro experiments, Ha.Cr demonstrated a concentration-dependent (0.25–4 mg/ml) inhibitory effect on the slope of aggregation. It decreased the size of crystals and transformed the calcium oxalate monohydrate (COM) to calcium oxalate dehydrate (COD) crystals, in calcium oxalate metastable solutions. It also showed concentration-dependent antioxidant effect against 2,2-diphenyl-1-picrylhydrazyl free radical (DPPH) free radicals and lipid peroxidation induced in rat kidney tissue homogenate. Ha.Cr (0.3 mg/ml) reduced (p < 0.05) the cell toxicity and LDH release in renal epithelial cells (MDCK) exposed to oxalate (0.5 mM) and COM (66 ?g/cm2) crystals. In male Wistar rats, receiving 0.75% ethylene glycol (EG) for 21 days along with 1% ammonium chloride (AC) in drinking water, Ha.Cr treatment (30–100 mg/kg) prevented the toxic changes caused by lithogenic agents; EG and AC, like loss of body weight, polyurea, oxaluria, raised serum urea and creatinine levels and crystal deposition in kidneys compared to their respective controls. Conclusion These data indicate that Holarrhena antidysenterica possesses antiurolithic activity, possibly mediated through inhibition of CaOx crystal aggregation, antioxidant and renal epithelial cell protective activities and may provide base for designing future studies to establish its efficacy and safety for clinical use. PMID:22622371

  19. Antioxidant activity and hepatoprotective potential of agaro-oligosaccharides in vitro and in vivo

    PubMed Central

    Chen, Haimin; Yan, Xiaojun; Zhu, Peng; Lin, Jing

    2006-01-01

    Background Agaro-oligosaccharides derived from red seaweed polysaccharide have been reported to possess antioxidant activity. In order to assess the live protective effects of agar-oligosaccharides, we did both in vitro and in vivo studies based on own-made agaro-oligosaccharides, and the structural information of this oligosaccharide was also determined. Method Structure of agaro-oligosaccharides prepared with acid hydrolysis on agar was confirmed by matrix-assisted ultraviolet laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) and NMR. The antioxidant effect of agaro-oligosaccharides on intracellular reactive oxygen species (ROS) was assessed by 2', 7'-dichlorofluorescin diacetate. Carbon tetrachloride was used to induce liver injury, some index including SOD, GSH-Px, MDA, AST, ALT were examined to determine the hepatoprotective effect of agaro-oligosaccharides. Results Agaro-oligosaccharides we got were composed of odd polymerizations with molecular weights ranged from 500 to 2500. Results from intracellular test indicated that agaro-oligosaccharides could significantly scavenge the level of oxidants in the hepatocytes, more beneficially, also associated with the improvement of cell viability In vivo studies of the antioxidant effects on tissue peroxidative damage induced by carbon tetrachloride in rat model indicated that agaro-oligosaccharides could elevate the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and decrease the level of malondialdehyde (MDA), glutamate oxaloacetate transaminase (AST), glutamic pyruvic transaminase (ALT) significantly. At 400 mg/kg, MDA level reduced 44 % and 21 % in liver and heart, SOD and GSH-Px increased to highest in liver and serum, while ALT level decreased 22.16 % in serum. Conclusion Overall, the results of the present study indicate that agaro-oligosaccharides can exert their in vitro and in vivo hepatoprotective effect through scavenging oxidative damage induced by ROS. PMID:17140450

  20. Assessment of the in vitro antimicrobial activity of Lactobacillus species for identifying new potential antibiotics.

    PubMed

    Dubourg, Grégory; Elsawi, Ziena; Raoult, Didier

    2015-11-01

    The bacteriocin-mediated antimicrobial properties of Lactobacillus spp. have been widely studied, leading to the use of these micro-organisms in the food industry as preservative agents against foodborne pathogens. In an era in which antibiotic resistance is becoming a public health issue, the antimicrobial activity of Lactobacillus spp. could be used for the discovery of new potential antibiotics. Thus, it is essential to have an accurate method of screening the production of antimicrobial agents by prokaryotes. Many in vitro assays have been published to date, largely concerning but not limited to Lactobacillus spp. However, these methods mainly use the spot-on-the-lawn method, which is prone to contamination during the overlay stage, with protocols using methanol vapours or the reverse side agar technique being applied to avoid such contamination. In this study, a method combining the spot-on-the-lawn and well diffusion methods was tested, permitting clear identification of inhibition zones from eight Lactobacillus spp. towards clinical isolates of 12 species (11 bacteria and 1 yeast) commonly found in human pathology. Lactobacillus plantarum CIP 106786 and Lactobacillus rhamnosus CSUR P567 exhibited the widest antimicrobial activity, whereas Lactobacillus acidophilus strain DSM 20079 was relatively inactive. In addition, the putative MIC50 of L. rhamnosus against Proteus mirabilis was estimated at 1.1×10(9)CFU/mL using culture broth dilution. In conclusion, considering the increasing cultivable bacterial human repertoire, these findings open the way of an effective method to screen in vitro for the production of potential antimicrobial compounds. PMID:26163158

  1. Antioxidant and antiproliferative activities of Abrus precatorius leaf extracts - an in vitro study

    PubMed Central

    2013-01-01

    Background The use of traditional medicine at the primary health care level is widespread and plant-based treatments are being recommended for curing various diseases by traditional medical practitioners all over the world. The phytochemicals present in the fruits, vegetables and medicinal plants are getting attention day-by-day for their active role in the prevention of several human diseases. Abrus precatorius is a widely distributed tropical medicinal plant with several therapeutic properties. Therefore in the present study, A. precatorius leaf extracts were examined for their antioxidant and cytotoxic properties in vitro in order to discover resources for new lead structures or to improve the traditional medicine. Methods In this study, antioxidant and antiproliferative properties of the different leaf extracts (hexane, ethyl acetate, ethanol and water) from A. precatorius were investigated along with the quantification of the polyphenol and flavonoid contents. The ability of deactivating free radicals was extensively investigated with in vitro biochemical methods like DPPH•, •OH, NO, SO2- scavenging assays and inhibition capability of Fe(II)-induced lipid peroxidation. Furthermore, antiproliferative activities using different human cancer cell lines and primary cell line was carried out by MTT method. Results Total phenolic content and total flavonoid content of the extracts were found in the range of 1.65?±?0.22 to 25.48?±?0.62 GAE mg/g dw and 6.20?±?0.41 to 17.16?±?1.04 QE mg/g dw respectively. The experimental results further revealed that A. precatorius extracts showed strong antiradical properties, capable to chelate Fe2+ and possess good inhibition ability of lipid peroxidation. In addition, as a first step towards the identification of phytoconstituents endowed with potent chemopreventive activities, we evaluated the inhibitory effects of A. precatorius extracts on the proliferation of four different human tumour cell lines such as human colon adenocarcinoma cells (Colo-205), human retinoblastoma cancer cells (Y79), human hepatocellular carcinoma cells (HepG2) and Leukemia cells (SupT1). Ethanol extract (APA) and ethyl acetate extract (APE) of A. precatorius had apparent capabilities of inhibiting the survival of tested human cancer cell lines. Moreover, it was observed that the A. precatorius extracts did not inhibit the growth of mice peritoneal macrophages, thus confirming that plants extracts are selective against the cancer cell lines. Conclusion This work provides a scientific support for the high antioxidant and antiproliferative activity of this plant and thus it may find potential applications in the treatment of the diseases caused by ROS. Further studies are needed to confirm in vivo anti-tumorgenicity and subsequent chemical characterization of the active molecule(s). PMID:23452983

  2. In vitro biological screening of the anticholinesterase and antiproliferative activities of medicinal plants belonging to Annonaceae

    PubMed Central

    Formagio, A.S.N.; Vieira, M.C.; Volobuff, C.R.F.; Silva, M.S.; Matos, A.I.; Cardoso, C.A.L.; Foglio, M.A.; Carvalho, J.E.

    2015-01-01

    The aim of this research was to investigate the antiproliferative and anticholinesterase activities of 11 extracts from 5 Annonaceae species in vitro. Antiproliferative activity was assessed using 10 human cancer cell lines. Thin-layer chromatography and a microplate assay were used to screen the extracts for acetylcholinesterase (AchE) inhibitors using Ellman's reagent. The chemical compositions of the active extracts were investigated using high performance liquid chromatography. Eleven extracts obtained from five Annonaceae plant species were active and were particularly effective against the UA251, NCI-470 lung, HT-29, NCI/ADR, and K-562 cell lines with growth inhibition (GI50) values of 0.04-0.06, 0.02-0.50, 0.01-0.12, 0.10-0.27, and 0.02-0.04 µg/mL, respectively. In addition, the Annona crassiflora and A. coriacea seed extracts were the most active among the tested extracts and the most effective against the tumor cell lines, with GI50 values below 8.90 µg/mL. The A. cacans extract displayed the lowest activity. Based on the microplate assay, the percent AchE inhibition of the extracts ranged from 12 to 52%, and the A. coriacea seed extract resulted in the greatest inhibition (52%). Caffeic acid, sinapic acid, and rutin were present at higher concentrations in the A. crassiflora seed samples. The A. coriacea seeds contained ferulic and sinapic acid. Overall, the results indicated that A. crassiflora and A. coriacea extracts have antiproliferative and anticholinesterase properties, which opens up new possibilities for alternative pharmacotherapy drugs. PMID:25714885

  3. In vitro biological screening of the anticholinesterase and antiproliferative activities of medicinal plants belonging to Annonaceae.

    PubMed

    Formagio, A S N; Vieira, M C; Volobuff, C R F; Silva, M S; Matos, A I; Cardoso, C A L; Foglio, M A; Carvalho, J E

    2015-04-01

    The aim of this research was to investigate the antiproliferative and anticholinesterase activities of 11 extracts from 5 Annonaceae species in vitro. Antiproliferative activity was assessed using 10 human cancer cell lines. Thin-layer chromatography and a microplate assay were used to screen the extracts for acetylcholinesterase (AchE) inhibitors using Ellman's reagent. The chemical compositions of the active extracts were investigated using high performance liquid chromatography. Eleven extracts obtained from five Annonaceae plant species were active and were particularly effective against the UA251, NCI-470 lung, HT-29, NCI/ADR, and K-562 cell lines with growth inhibition (GI50) values of 0.04-0.06, 0.02-0.50, 0.01-0.12, 0.10-0.27, and 0.02-0.04 µg/mL, respectively. In addition, the Annona crassiflora and A. coriacea seed extracts were the most active among the tested extracts and the most effective against the tumor cell lines, with GI50 values below 8.90 µg/mL. The A. cacans extract displayed the lowest activity. Based on the microplate assay, the percent AchE inhibition of the extracts ranged from 12 to 52%, and the A. coriacea seed extract resulted in the greatest inhibition (52%). Caffeic acid, sinapic acid, and rutin were present at higher concentrations in the A. crassiflora seed samples. The A. coriacea seeds contained ferulic and sinapic acid. Overall, the results indicated that A. crassiflora and A. coriacea extracts have antiproliferative and anticholinesterase properties, which opens up new possibilities for alternative pharmacotherapy drugs. PMID:25714885

  4. Trans-cleaving hammerhead ribozymes with tertiary stabilizing motifs: in vitro and in vivo activity against a structured viroid RNA

    PubMed Central

    Carbonell, Alberto; Flores, Ricardo; Gago, Selma

    2011-01-01

    Trans-cleaving hammerheads with discontinuous or extended stem I and with tertiary stabilizing motifs (TSMs) have been tested previously against short RNA substrates in vitro at low Mg2+ concentration. However, the potential of these ribozymes for targeting longer and structured RNAs in vitro and in vivo has not been examined. Here, we report the in vitro cleavage of short RNAs and of a 464-nt highly structured RNA from potato spindle tuber viroid (PSTVd) by hammerheads with discontinuous and extended formats at submillimolar Mg2+. Under these conditions, hammerheads derived from eggplant latent viroid and peach latent mosaic viroid (PLMVd) with discontinuous and extended formats, respectively, where the most active. Furthermore, a PLMVd-derived hammerhead with natural TSMs showed activity in vivo against the same long substrate and interfered with systemic PSTVd infection, thus reinforcing the idea that this class of ribozymes has potential to control pathogenic RNA replicons. PMID:21097888

  5. In vitro and in vivo aldose reductase inhibitory activity of standardized extracts and the major constituent of Andrographis paniculata.

    PubMed

    Veeresham, Ciddi; Swetha, Ettireddy; Rao, Ajmeera Rama; Asres, Kaleab

    2013-03-01

    Aldose reductase is the first enzyme in the polyol pathway and catalyzes the reduction of glucose to sorbitol by coupling with the oxidation of NADPH to NADP(+) . This sorbitol accumulation leads to various diabetic complications, including neuropathy, nephropathy, cataracts, and retinopathy. In the present study, aldose reductase inhibitory (ARI) activity of the methanolic as well as standardized extracts of Andrographis paniculata (Burm. f.) Wall. ex Nees (Acanthaceae) and its chief constituent, andrographolide, were studied using in vitro and in vivo methods. In the in vitro method, rat lens as well as kidney homogenates were used for the preparation of enzyme, whereas the effect of these test samples on the galactitol level in the eye lens was studied in a galactosemic rat model in vivo. The results of the study revealed that both extracts of the plant and its major compound, andrographolide, possess ARI activity in vitro. They were also found to significantly decrease galactitol accumulation in vivo. PMID:22628202

  6. Oxiconazole, a new imidazole derivative. Evaluation of antifungal activity in vitro and in vivo.

    PubMed

    Polak, A

    1982-01-01

    The new imidazole derivative Z-[2,4-dichloro-2-imidazol-1-yl)acetophenone]-O-(2,4-dichlorobenzyl)-oxime nitrate (oxiconazole, Ro 13-8996) is characterized by a broad fungistatic spectrum against the agents of human mycoses in vitro. In addition, fungicidal activity of various degree was found in selected species (Aspergillus fumigatus, Cryptococcus neoformans, Candida albicans and Trichophyton mentagrophytes). Synthesis of DNA was inhibited by subinhibitory concentrations of oxiconazole in parallel to cell multiplication, whereas synthesis of RNA, protein and carbohydrate was decreased to a lesser extent. The most relevant findings was high topical activity in both trichophytosis in the guinea-pig and vaginal candidiasis in the rat. In these 2 models, oxiconazole proved to be more potent than several reference compounds from the group of imidazole antimycotics. Systemic oral activity of oxiconazole was also found in three mouse models, namely in dermal infection with T. mentagrophytes var. quinckeanum, systemic histoplasmosis and, just to a low degree, systemic candidiasis, but the compound proved to be inactive in cryptococcosis and aspergillosis in the mouse. Based on our findings, a clinical evaluation of oxiconazole as a topical antimycotic in human superficial mycosis, seems to be justified. PMID:7037014

  7. Hypocholesterolemic Effect and In Vitro Pancreatic Lipase Inhibitory Activity of an Opuntia ficus-indica Extract

    PubMed Central

    Flores-Fernandez, Jose Miguel; Fernandez-Flores, Ofelia; Gutierrez-Mercado, Yanet; Carmona-de la Luz, Joel; Sandoval-Salas, Fabiola; Mendez-Carreto, Carlos

    2015-01-01

    Cholesterol control is fundamental for prevention of cardiovascular disorders. In this work, the hypocholesterolemic activity of an aqueous Opuntia ficus-indica extract (AOE) was tested in triton-induced mice. The inhibitory activity on pancreatic lipase enzyme was evaluated in vitro by the same extract. Furthermore, polyphenol content of the extract was evaluated. Hypercholesterolemia was induced in three groups of mice by intraperitoneal administration of Triton WR-1339. After induction of hypercholesterolemia, the groups were treated with an AOE (500?mg/kg) and saline solution and the positive control group with orlistat, respectively. Cholesterol levels were measured 24?h later in peripheral blood. The levels of blood cholesterol after administration of AOE significantly decreased compared to negative control. The inhibitory activity of AOE on pancreatic lipase enzyme was evaluated at concentrations from 60 to 1000??g/mL. The AOE inhibited the pancreatic lipase with an IC50 = 588.5??g/mL. The AOE had a high content of polyphenolic compounds. These results show that AOE is able to prevent hypercholesterolemia by pancreatic lipase inhibition, in part due to its polyphenolic compounds. PMID:26078966

  8. Telavancin: mechanisms of action, in vitro activity, and mechanisms of resistance.

    PubMed

    Karlowsky, James A; Nichol, Kim; Zhanel, George G

    2015-09-15

    Telavancin is a semisynthetic lipoglycopeptide derivative of vancomycin. Telavancin has a dual mechanism of antibacterial action, disrupting peptidoglycan synthesis and cell membrane function. In 2014, the Clinical and Laboratory Standards Institute (CLSI) revised the antimicrobial susceptibility testing method for telavancin, resulting in minimum inhibitory concentration (MIC) determinations that are more accurate and reproducible and demonstrate greater in vitro potency than shown with the previous testing method. The CLSI testing method changes coincided with revised telavancin MIC interpretive break point criteria for susceptibility approved by the US Food and Drug Administration for Staphylococcus aureus (?0.12 µg/mL), Streptococcus pyogenes (?0.12 µg/mL), Streptococcus agalactiae (?0.12 µg/mL), Streptococcus anginosus group (?0.06 µg/mL), and Enterococcus faecalis (vancomycin susceptible, ?0.25 µg/mL). Telavancin is equally potent against methicillin-susceptible S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA). It demonstrates activity against isolates of heterogeneous vancomycin-intermediate S. aureus and vancomycin-intermediate S. aureus but is poorly active against vancomycin-resistant S. aureus. It also demonstrates potent activity against Staphylococcus epidermidis and Streptococcus spp. (MIC90 ?0.03 µg/mL). Thus far, it has not been possible to select for high-level telavancin resistance in the laboratory using serially passaged clinical isolates of MRSA and MSSA. PMID:26316559

  9. Optimization extraction, preliminary characterization and antioxidant activity in vitro of polysaccharides from Stachys sieboldii Miq. tubers.

    PubMed

    Feng, Kai; Chen, Wei; Sun, Liwei; Liu, Jianzeng; Zhao, Yangxin; Li, Luxi; Wang, Yuxing; Zhang, Wenjing

    2015-07-10

    Response surface methodology was used to optimize the extraction conditions of water-soluble polysaccharides from Stachys sieboldii Miq. tubers. A central composite design was used to optimize the extraction processing parameters. The optimum extraction conditions are as follows: extraction temperature, 95°C; extraction time, 2.5h; water to raw material ratio, 16; and extraction frequency, 3. Under the optimized conditions, an experimental yield of 9.21 ± 0.18%, which is in good agreement with the predicted yield, was obtained. Purified polysaccharide SSP II-a was successfully obtained using diethylaminoethanol-Sepharose and Sepharose CL-6B column chromatography. SSP II-a was found to be an acidic polysaccharide fraction with an average molecular weight of 168kDa and composed of rhamnose, glucuronic acid, galacturonic acid, glucose, galactose and arabinose. In vitro antioxidant activity assays suggested that SSP II-a presents high scavenging activity toward superoxide anion, hydroxyl, and 2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) radicals but relatively lower scavenging activity toward 1,1-diphenyl-2-picrylhydrazyl radicals. The results indicated that response surface methodology is an effective method for the extraction of polysaccharides from S. sieboldii Miq. tubers and the polysaccharides could be explored as a potential antioxidant agent for use in medicine or functional food. PMID:25857958

  10. Environmental gestagens activate fathead minnow (Pimephales promelas) nuclear progesterone and androgen receptors in vitro.

    PubMed

    Ellestad, Laura E; Cardon, Mary; Chambers, Ian G; Farmer, Jennifer L; Hartig, Phillip; Stevens, Kyle; Villeneuve, Daniel L; Wilson, Vickie; Orlando, Edward F

    2014-07-15

    Gestagen is a collective term for endogenous and synthetic progesterone receptor (PR) ligands. In teleost fishes, 17?,20?-dihydroxy-4-pregnen-3-one (DHP) and 17?,20?,21-trihydroxy-4-pregnen-3-one (20?-S) are the predominant progestogens, whereas in other vertebrates the major progestogen is progesterone (P4). Progestins are components of human contraceptives and hormone replacement pharmaceuticals and, with P4, can enter the environment and alter fish and amphibian reproductive health. In this study, our primary objectives were to clone the fathead minnow (FHM) nuclear PR (nPR), to develop an in vitro assay for FHM nPR transactivation, and to screen eight gestagens for their ability to transactivate FHM nPR. We also investigated the ability of these gestagens to transactivate FHM androgen receptor (AR). Fish progestogens activated FHM nPR, with DHP being more potent than 20?-S. The progestin drospirenone and P4 transactivated the FHM nPR, whereas five progestins and P4 transactivated FHM AR, all at environmentally relevant concentrations. Progestins are designed to activate human PR, but older generation progestins have unwanted androgenic side effects in humans. In FHMs, several progestins proved to be strong agonists of AR. Here, we present the first mechanistic evidence that environmental gestagens can activate FHM nPR and AR, suggesting that gestagens may affect phenotype through nPR- and AR-mediated pathways. PMID:24911891

  11. Guarana (Paullinia cupana): toxic behavioral effects in laboratory animals and antioxidants activity in vitro.

    PubMed

    Mattei, R; Dias, R F; Espínola, E B; Carlini, E A; Barros, S B

    1998-03-01

    The effects on toxic and behavioral levels of guarana (Paullinia cupana) were assessed in rats and mice subsequent to acute and chronic administrations and were compared to those produced by Ginseng (Panax ginseng). Experimental parameters included tests for antioxidant capacity in vitro and measured in vivo, toxicological screening, progress in weight, motor activity, death rate, and histopathological examination of the viscera. Guarana showed an antioxidant effect because, even at low concentrations (1.2 microg/ml), it inhibited the process of lipid peroxidation. In high doses of 1000-2000 mg/kg (i.p. and p.o.) it did not induce significant alterations in parameters for toxicological screening. No effects on motor activity were observed, neither did guarana alter the hypnotic effect of pentobarbital. Ginseng (250-1000 mg/kg i.p.), however, elicited reductions in motor activity, eyelid ptosis and bristling fur. Consumption of liquids containing guarana or ginseng and progress in weight of the animals remained at levels similar to the controls, even after prolonged administration. The percentage mortality was equivalent in control and in treated groups. The absence of toxicity of guarana was also demonstrated by histopathological examination, with no alteration being detected in heart, lungs, stomach, small and large intestine, liver, pancreas, kidneys, bladder and spleen. PMID:9582000

  12. In vitro evaluation of antimicrobial activity of sealers and pastes used in endodontics.

    PubMed

    Leonardo, M R; da Silva, L A; Tanomaru Filho, M; Bonifácio, K C; Ito, I Y

    2000-07-01

    The antimicrobial activity of four root canal sealers (AH Plus, Sealapex, Ketac Endo, and Fill Canal), two calcium hydroxide pastes (Calen and Calasept), and a zinc oxide paste was evaluated. Seven bacterial strains were used, six of them standard; Micrococcus luteus ATCC 9341, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Staphylococcus epidermidis ATCC 12228, Escherichia coli ATCC 25922, and Enterococcus faecalis ATCC 10541. There was a wild strain of Streptococcus mutans isolated from saliva obtained in an adult dental clinic. Activity was evaluated using the agar diffusion method with Brain Heart Infusion agar and Müller Hinton medium seeded by pour plate. Calcium hydroxide-based sealers and pastes were either placed directly into 4.0 x 4.0 mm wells or by using absorbent paper points. The plates were kept at room temperature for 2 hr for diffusion. After incubation at 37 degrees C for 24 hr, the medium was optimized with 0.05 g% TTC gel and inhibition haloes were measured. All bacterial strains were inhibited by all materials using the well method. However, when the materials were applied with absorbent paper points, Enterococcus faecalis was not inhibited by zinc oxide, and Pseudomonas aeruginosa was not inhibited by AH Plus, Fill Canal, and the zinc oxide-based paste. We conclude that sealers and pastes presented antimicrobial activity in vitro and culture medium optimization with 0.05 g% TTC gel facilitated observation of the inhibition haloes. PMID:11199762

  13. Comparative in vitro activity of tigecycline against bacteria recovered from clinical specimens in Latin America.

    PubMed

    Bantar, C; Curcio, D; Fernandez Canigia, L; García, P; Guzmán Blanco, M; Leal, A L

    2009-04-01

    The present study was performed to evaluate the in vitro activity of tigecycline in comparison to other agents against isolates recovered from patients hospitalized in latin American. Organisms were collected in 47 clinical laboratories from 4 countries of latin America between November 2005 and October 2006 and were tested by using disk diffusion method as described by the CLSI. A total of 7966 isolates were assessed. Tigecycline proved highly active against staphylococci and enterococci (>99% susceptibility). Imipenem was the most active agent against Escherichia coli (100% susceptibility), followed by tigecycline, 98.6% susceptibility. Resistance to cefotaxime in this species was 15.3%. Global tigecycline susceptibility of Klebsiella species was 90.2%, but the susceptibility rate was significantly slower in Venezuela (82%) than in Argentina, Colombia and Chile (93%) (p<0.01). Global cefotaxime resistance to Klebsiella spp. was 32.2% and carbapenem resistance was detected in all countries. By adopting a susceptible breakpoint >or =16mm, 91.3% of the Acinetobacter isolates proved susceptible to tigecycline. Results from the present study suggest that tigecycline may be a suitable option in latin America, a region where multidrug resistance seems to be a dramatic, increasing problem and new antimicrobial choices are urgently needed. PMID:19423466

  14. Monitoring Hippocampus Electrical Activity In Vitro on an Elastically Deformable Microelectrode Array

    PubMed Central

    Yu, Zhe; Graudejus, Oliver; Tsay, Candice; Lacour, Stéphanie P.; Wagner, Sigurd

    2009-01-01

    Abstract Interfacing electronics and recording electrophysiological activity in mechanically active biological tissues is challenging. This challenge extends to recording neural function of brain tissue in the setting of traumatic brain injury (TBI), which is caused by rapid (within hundreds of milliseconds) and large (greater than 5% strain) brain deformation. Interfacing electrodes must be biocompatible on multiple levels and should deform with the tissue to prevent additional mechanical damage. We describe an elastically stretchable microelectrode array (SMEA) that is capable of undergoing large, biaxial, 2-D stretch while remaining functional. The new SMEA consists of elastically stretchable thin metal films on a silicone membrane. It can stimulate and detect electrical activity from cultured brain tissue (hippocampal slices), before, during, and after large biaxial deformation. We have incorporated the SMEA into a well-characterized in vitro TBI research platform, which reproduces the biomechanics of TBI by stretching the SMEA and the adherent brain slice culture. Mechanical injury parameters, such as strain and strain rate, can be precisely controlled to generate specific levels of damage. The SMEA allowed for quantification of neuronal function both before and after injury, without breaking culture sterility or repositioning the electrodes for the injury event, thus enabling serial and long-term measurements. We report tests of the SMEA and an initial application to study the effect of mechanical stimuli on neuron function, which could be employed as a high-content, drug-screening platform for TBI. PMID:19594385

  15. In vitro antibacterial activity of Hibiscus rosa-sinensis flower extract against human pathogens

    PubMed Central

    Ruban, P; Gajalakshmi, K

    2012-01-01

    Objective To access the in vitro antibacterial activity of Hibiscus rosa-sinensis (H. rosa- sinensis) flower extract against human pathogens. Methods Antibacterial activity was evaluated by using disc and agar diffusion methods. The protein was run through poly acrylmide gel electrophoresis to view their protein profile. Results The results showed that the cold extraction illustrates a maximum zone of inhibition against Bacillus subtillis (B. subtillis), Escherichia coli (E. coli) viz., (17.00 ± 2.91), (14.50 ± 1.71) mm, followed by hot extraction against, E. coli, Salmonella sp. as (11.66 ± 3.14), (10.60 ± 3.09) mm. In methanol extraction showed a highest zone of inhibition recorded against B. subtillis, E. coli as (18.86 ± 0.18), (18.00 ± 1.63) mm pursued by ethanol extraction showed utmost zone of inhibition recorded against Salmonella sp. at (20.40 ± 1.54) mm. The crude protein from flower showed a maximum inhibitory zone observed against Salmonella sp., E. coli viz., (16.55 ± 1.16), (14.30 ± 2.86) mm. The flower material can be taken as an alternative source of antibacterial agent against the human pathogens. Conclusions The extracts of the H. rosa-sinensis are proved to have potential antibacterial activity, further studies are highly need for the drug development. PMID:23569938

  16. Acaricidal activity of extracts from Ligularia virgaurea against the Sarcoptes scabiei mite in vitro

    PubMed Central

    LUO, BIAO; LIAO, FEI; HU, YANCHUN; LIU, XI; HE, YAJUN; WU, LEI; TAN, HUI; LUO, LIJUAN; ZHOU, YANCHENG; MO, QUAN; DENG, JUNLIANG; WEI, YAHUI

    2015-01-01

    The aim of the present study was to investigate the acaricidal activity of Ligularia virgaurea. An extract was prepared by refluxing with ethanol and steam distillation, and its toxic effect was tested in vitro against Sarcoptes scabiei. The data from the toxicity tests were analyzed using a complementary log-log (CLL) model. The ethanol extract exhibited strong acaricidal activity against these mites; at a concentration of 2 g/ml it killed all S. scabiei within 2 h and at 1 g/ml it killed all S. scabiei within 6 h. Similarly, 2, 1, 0.5 and 0.25 g/ml concentrations of the extract had strong toxicity against S. scabiei, with median lethal time (LT50) values of 0.716, 1.741, 2.968 and 4.838 h, respectively. The median lethal concentration (LC50) values were 1.388, 0.624, 0.310 and 0.213 g/ml for Scabies mite in 1, 2, 4 and 6 h, respectively. The results indicate that the L. virgaurea extract has strong acaricidal activity and may be exploited as a novel treatment for the effective control of acariasis in animals. PMID:26170943

  17. In vitro antibacterial activity of seven Indian spices against high level gentamicin resistant strains of enterococci

    PubMed Central

    Bipin, Chapagain; Chitra, Pai (Bhat); Minakshi, Bhattacharjee

    2015-01-01

    Introduction The aim of the study was to explore the in vitro antibacterial activity of seven ethanolic extracts of spices against high level gentamicin resistant (HLGR) enterococci isolated from human clinical samples. Material and methods Two hundred and fifteen enterococcal strains were isolated from clinical samples. High level gentamicin resistance in ethanolic extracts of cumin (Cuminum cyminum), cinnamon (Cinnamomum zeylanicum), ginger (Zingiber officinale), fenugreek (Trigonella foenum-graecum), cloves (Syzygium aromaticum), cardamom (Elettaria cardamomum Maton) and black pepper (Piper nigrum) were prepared using Soxhlet apparatus. The antibacterial effect of the extracts was studied using the well diffusion method. Statistical analysis was carried out by ?2 test using SPSS 17 software. Results Only cinnamon and ginger were found to have activity against all the isolates, whereas cumin and cloves had a variable effect on the strains. Fenugreek, black pepper and cardamom did not show any effect on the isolates. The zone diameter of inhibition obtained for cinnamon, ginger, cloves and cumin was in the range 31–34 mm, 27–30 mm, 25–26 mm and 19–20 mm respectively. Conclusions Cinnamomum zeylanicum and Z. officinale showed the maximum antibacterial activity against the enterococcal isolates followed by S. aromaticum and C. cyminum. The findings of the study show that spices used in the study can contribute to the development of potential antimicrobial agents for inclusion in the anti-enterococcal treatment regimen. PMID:26322099

  18. In vitro activity of synthetic tetrahydroindeno[2,1-c]quinolines on Leishmania mexicana.

    PubMed

    Hernández-Chinea, Concepción; Carbajo, Erika; Sojo, Felipe; Arvelo, Francisco; Kouznetsov, Vladimir V; Romero-Bohórquez, Arnold R; Romero, Pedro J

    2015-12-01

    New synthetic compounds based on tetrahydroindenoquinoline structure were evaluated for their in vitro antileishmanial activities. The seven compounds assayed have antiproliferative activities against promastigotes of Leishmania mexicana. Compound 1 and 3 were the most active (IC50 1.0 ?g/ml) and showed high selectivity towards the parasite. These compounds were selected to evaluate their effect on promastigote morphology and mitochondrial transmembrane potential as well as on the amastigote capability to survive into macrophages J774 cell line. Whereas compound 1 affected the promastigote cell cycle, compound 3 induced morphological changes and the total collapse of the mitochondrial transmembrane potential, a hallmark of apoptosis. Both compounds also affected the amastigote form of the parasite, decreasing their survival rate in J774 macrophages. Due to the greatest selectivity index, the apparent effect as apoptotic inducer and its sustained inhibition on intracellular amastigote replication, compound 3 is the best candidate to be tested in vivo. This compound is worth considering for the development of new antileishmanial drugs. PMID:26148815

  19. In Vitro Antiprotozoal Activity of Abietane Diterpenoids Isolated from Plectranthus barbatus Andr.

    PubMed Central

    Mothana, Ramzi A.; Al-Said, Mansour S.; Al-Musayeib, Nawal M.; El Gamal, Ali A.; Al-Massarani, Shaza M.; Al-Rehaily, Adnan J.; Abdulkader, Majed; Maes, Louis

    2014-01-01

    Chromatographic separation of the n-hexane extract of the aerial part of Plectranthus barbatus led to the isolation of five abietane-type diterpenes: dehydroabietane (1); 5,6-didehydro-7-hydroxy-taxodone (2); taxodione (3); 20-deoxocarnosol (4) and 6?,11,12,-trihydroxy-7?,20-epoxy-8,11,13-abietatriene (5). The structures were determined using spectroscopic methods including one- and two-dimensional NMR methods. Compounds (1)–(3) and (5) are isolated here for the first time from the genus Plectranthus. The isolated abietane-type diterpenes tested in vitro for their antiprotozoal activity against erythrocytic schizonts of Plasmodium falciparum, intracellular amastigotes of Leishmania infantum and Trypanosoma cruzi and free trypomastigotes of T. brucei. Cytotoxicity was determined against fibroblast cell line MRC-5. Compound (2) 5,6-didehydro-7-hydroxy-taxodone showed remarkable activity with acceptable selectivity against P. falciparum (IC50 9.2 ?M, SI 10.4) and T. brucei (IC50 1.9 ?M, SI 50.5). Compounds (3)–(5) exhibited non-specific antiprotozoal activity due to high cytotoxicity. Compound (1) dehydroabietane showed no antiprotozoal potential. PMID:24823881

  20. Hypocholesterolemic Effect and In Vitro Pancreatic Lipase Inhibitory Activity of an Opuntia ficus-indica Extract.

    PubMed

    Padilla-Camberos, Eduardo; Flores-Fernandez, Jose Miguel; Fernandez-Flores, Ofelia; Gutierrez-Mercado, Yanet; Carmona-de la Luz, Joel; Sandoval-Salas, Fabiola; Mendez-Carreto, Carlos; Allen, Kirk

    2015-01-01

    Cholesterol control is fundamental for prevention of cardiovascular disorders. In this work, the hypocholesterolemic activity of an aqueous Opuntia ficus-indica extract (AOE) was tested in triton-induced mice. The inhibitory activity on pancreatic lipase enzyme was evaluated in vitro by the same extract. Furthermore, polyphenol content of the extract was evaluated. Hypercholesterolemia was induced in three groups of mice by intraperitoneal administration of Triton WR-1339. After induction of hypercholesterolemia, the groups were treated with an AOE (500?mg/kg) and saline solution and the positive control group with orlistat, respectively. Cholesterol levels were measured 24?h later in peripheral blood. The levels of blood cholesterol after administration of AOE significantly decreased compared to negative control. The inhibitory activity of AOE on pancreatic lipase enzyme was evaluated at concentrations from 60 to 1000??g/mL. The AOE inhibited the pancreatic lipase with an IC50 = 588.5??g/mL. The AOE had a high content of polyphenolic compounds. These results show that AOE is able to prevent hypercholesterolemia by pancreatic lipase inhibition, in part due to its polyphenolic compounds. PMID:26078966

  1. Free radical scavenging activity and comparative metabolic profiling of in vitro cultured and field grown Withania somnifera roots.

    PubMed

    Senthil, Kalaiselvi; Thirugnanasambantham, Pankajavalli; Oh, Taek Joo; Kim, So Hyun; Choi, Hyung Kyoon

    2015-01-01

    Free radical scavenging activity (FRSA), total phenolic content (TPC), and total flavonoid content (TFC) of in vitro cultured and field grown Withania somnifera (Ashwagandha) roots were investigated. Withanolides analysis and comprehensive metabolic profiling between 100% methanol extracts of in vitro and field grown root tissues was performed using high performance thin layer chromatography (HPTLC) and gas chromatography-mass spectrometry (GC-MS), respectively. Significantly higher levels of FRSA, TPC, and TFC were observed in in-vitro cultured roots compared with field grown samples. In addition, 30 day-cultured in vitro root samples (1 MIR) exhibited a significantly higher FRSA (IC50 81.01 ?g/mL), TPC (118.91 mg GAE/g), and TFC (32.68 mg CE/g) compared with those in 45 day-cultured samples (1.5 MIR). Total of 29 metabolites were identified in in vitro cultured and field grown roots by GC-MS analysis. The metabolites included alcohols, organic acids, purine, pyrimidine, sugars, and putrescine. Vanillic acid was only observed in the in vitro cultured root samples, and higher level of the vanillic acid was observed in 1 MIR when compared to 1.5 MIR. Therefore, it is suggested that 1 MIR might serve as an alternative to field grown roots for the development of medicinal and functional food products. PMID:25874568

  2. Free Radical Scavenging Activity and Comparative Metabolic Profiling of In Vitro Cultured and Field Grown Withania somnifera Roots

    PubMed Central

    Senthil, Kalaiselvi; Thirugnanasambantham, Pankajavalli; Oh, Taek Joo; Kim, So Hyun; Choi, Hyung Kyoon

    2015-01-01

    Free radical scavenging activity (FRSA), total phenolic content (TPC), and total flavonoid content (TFC) of in vitro cultured and field grown Withania somnifera (Ashwagandha) roots were investigated. Withanolides analysis and comprehensive metabolic profiling between 100% methanol extracts of in vitro and field grown root tissues was performed using high performance thin layer chromatography (HPTLC) and gas chromatography-mass spectrometry (GC-MS), respectively. Significantly higher levels of FRSA, TPC, and TFC were observed in in-vitro cultured roots compared with field grown samples. In addition, 30 day-cultured in vitro root samples (1MIR) exhibited a significantly higher FRSA (IC50 81.01 ?g/mL), TPC (118.91 mg GAE/g), and TFC (32.68 mg CE/g) compared with those in 45 day-cultured samples (1.5MIR). Total of 29 metabolites were identified in in vitro cultured and field grown roots by GC-MS analysis. The metabolites included alcohols, organic acids, purine, pyrimidine, sugars, and putrescine. Vanillic acid was only observed in the in vitro cultured root samples, and higher level of the vanillic acid was observed in 1MIR when compared to 1.5MIR. Therefore, it is suggested that 1MIR might serve as an alternative to field grown roots for the development of medicinal and functional food products. PMID:25874568

  3. In Vitro Activity of ASP2397 against Aspergillus Isolates with or without Acquired Azole Resistance Mechanisms.

    PubMed

    Arendrup, Maiken Cavling; Jensen, Rasmus Hare; Cuenca-Estrella, Manuel

    2015-01-01

    ASP2397 is a new compound with a novel and as-yet-unknown target different from that of licensed antifungal agents. It has activity against Aspergillus and Candida glabrata. We compared its in vitro activity against wild-type and azole-resistant A. fumigatus and A. terreus isolates with that of amphotericin B, itraconazole, posaconazole, and voriconazole. Thirty-four isolates, including 4 wild-type A. fumigatus isolates, 24 A. fumigatus isolates with alterations in CYP51A TR/L98H (5 isolates), M220 (9 isolates), G54 (9 isolates), and HapE (1 isolate), and A. terreus isolates (2 wild-type isolates and 1 isolate with an M217I CYP51A alteration), were analyzed. EUCAST E.Def 9.2 and CLSI M38-A2 MIC susceptibility testing was performed. ASP2397 MIC50 values (in milligrams per liter, with MIC ranges in parentheses) determined by EUCAST and CLSI were 0.5 (0.25 to 1) and 0.25 (0.06 to 0.25) against A. fumigatus CYP51A wild-type isolates and were similarly 0.5 (0.125 to >4) and 0.125 (0.06 to >4) against azole-resistant A. fumigatus isolates, respectively. These values were comparable to those for amphotericin B, which were 0.25 (0.125 to 0.5) and 0.25 (0.125 to 0.25) against wild-type isolates and 0.25 (0.125 to 1) and 0.25 (0.125 to 1) against isolates with azole resistance mechanisms, respectively. In contrast, MICs for the azole compounds were elevated and highest for itraconazole: >4 (1 to >4) and 4 (0.5 to >4) against isolates with azole resistance mechanisms compared to 0.125 (0.125 to 0.25) and 0.125 (0.06 to 0.25) against wild-type isolates, respectively. ASP2397 was active against A. terreus CYP51A wild-type isolates (MIC 0.5 to 1), whereas MICs of both azole and ASP2397 were elevated for the mutant isolate. ASP2397 displayed in vitro activity against A. fumigatus and A. terreus isolates which was independent of the presence or absence of azole target gene resistance mutations in A. fumigatus. The findings are promising at a time when azole-resistant A. fumigatus is emerging globally. PMID:26552973

  4. Synthesis, in vitro biological activities and in silico study of dihydropyrimidines derivatives.

    PubMed

    Barakat, Assem; Islam, Mohammad Shahidul; Al-Majid, Abdullah Mohammed; Ghabbour, Hazem A; Fun, Hoong-Kun; Javed, Kulsoom; Imad, Rehan; Yousuf, Sammer; Choudhary, M Iqbal; Wadood, Abdul

    2015-10-15

    We describe here the synthesis of dihydropyrimidines derivatives 3a-p, and evaluation of their ?-glucosidase enzyme inhibition activities. Compounds 3b (IC50=62.4±1.5?M), 3c (IC50=25.3±1.26?M), 3d (IC50=12.4±0.15?M), 3e (IC50=22.9±0.25?M), 3g (IC50=23.8±0.17?M), 3h (IC50=163.3±5.1?M), 3i (IC50=30.6±0.6?M), 3m (IC50=26.4±0.34?M), and 3o (IC50=136.1±6.63?M) were found to be potent ?-glucosidase inhibitors in comparison to the standard drug acarbose (IC50=840±1.73?M). The compounds were also evaluated for their in vitro cytotoxic activity against PC-3, HeLa, and MCF-3 cancer cell lines, and 3T3 mouse fibroblast cell line. All compounds were found to be non cytotoxic, except compounds 3f and 3m (IC50=17.79±0.66-20.44±0.30?M), which showed a weak cytotoxic activity against the HeLa, and 3T3 cell lines. In molecular docking simulation study, all the compounds were docked into the active site of the predicted homology model of ?-glucosidase enzyme. From the docking result, it was observed that most of the synthesized compounds showed interaction through carbonyl oxygen atom and polar phenyl ring with active site residues of the enzyme. PMID:26381063

  5. In vitro antiprotozoal activity of extract and compounds from the stem bark of Combretum molle.

    PubMed

    Asres, K; Bucar, F; Knauder, E; Yardley, V; Kendrick, H; Croft, S L

    2001-11-01

    The antiprotozoal activity of the Ethiopian medicinal plant Combretum molle (R. Br. ex G. Don.) Engl & Diels (Combretaceae) was evaluated by in vitro testing against Plasmodium falciparum, Trypanosoma brucei rhodesiense, Trypanosoma cruzi and Leishmania donovani. The acetone fraction of the stem bark of this plant prepared by soxhlet extraction was inactive against the intracellular amastigotes of L. donovani and T. cruzi in murine peritoneal macrophages but showed significant activity against extracellular T. b. rhodesiense blood stream form trypomastigotes and trophozoites of P. falciparum with IC(50) values of 2.19 and 8.17 microg/mL, respectively. Phytochemical examination of the bioactive fraction resulted in the isolation of two tannins and two oleanane-type pentacyclic triterpene glycosides. One of the tannins was identified as the ellagitannin, punicalagin, whilst the structure of the other (CM-A) has not yet been fully elucidated. The saponins that were characterized as arjunglucoside (also called 4-epi-sericoside) and sericoside displayed no activity against any of the four species of protozoa tested. On the other hand, punicalagin and CM-A had IC(50) values of 1.75 and 1.50 microM, respectively, against T. b. rhodesiense and were relatively less toxic to KB cells (cytotoxic/antiprotozoal ratios of 70 and 48, respectively). The tannins also showed intermediate activity against P. falciparum, although their selectivity against these parasites was less favourable than the above. It appears that our findings are the first report of hydrolysable tannins exhibiting antitrypanosomal and antiplasmodial activities. PMID:11746844

  6. [In vitro activities of levofloxacin and other antibiotics against fresh clinical isolates].

    PubMed

    Matsuzaki, K; Koyama, H; Chiba, A; Omika, K; Harada, S; Sato, Y; Hasegawa, M; Kobayashi, I; Kaneko, A; Sasaki, J

    1999-09-01

    In this study, the in vitro activity of levofloxacin (LVFX) against 1,020 fresh bacterial clinical isolates was compared with the activities of a range of ofloxacin, ciprofloxacin (CPFX), ampicillin (ABPC), cefaclor, cefpodoxime, methicillin and benzylpenicillin. The clinical isolates except Vibrio cholerae were collected in Japan during 1998 from patients with infectious diseases. MICs were determined using the agar dilution method according to the recommendations by the Japan Society of Chemotherapy. Some isolates of methicillin resistant Staphylococcus aureus (MRSA) and coagulase negative Staphylococcus were resistant to fluoroquinolones, but the MIC50 of LVFX against MRSA was 6.25 micrograms/ml. LVFX was the most active against MRSA among the antibiotics tested. Most of Staphylococcus epidermidis strains were susceptible to the fluoroquinolones. LVFX showed greater activity against all streptococci strains compared with fluoroquinolones tested. In particular, all Streptococcus pneumoniae strains including PRSP were susceptible to LVFX at < or = 1.56 micrograms/ml. Among Enterococcus, ABPC showed superior activity against Enterococcus faecalis but many isolates of Enterococcus species were resistant to ABPC. LVFX was more active against to these Enterococcus species compared with other fluoroquinolones. On the other hand, LVFX and CPFX showed similar activity against isolates of Enterobacteriaceae. CPFX had an MIC50/90 of 0.20, 0.39 microgram/ml and LVFX showed an MIC50/90 of 0.78, 1.56 micrograms/ml against Pseudomonas aeruginosa. LVFX (MIC50/90 0.10, 0.20 microgram/ml) was more active against Acinetobacter species than CPFX (MIC50/90 0.10, 0.39 microgram/ml). Haemophilus influenzae, Branhamella (Moraxella) catarrhalis and V. cholerae were inhibited by low concentration of the fluoroquinolones tested. The MIC90 of LVFX and CPFX were < or = 0.10 microgram/ml against above three species. Some isolates of Neisseria gonorrhoeae and Campylobacter species were moderately resistant to the fluoroquinolones tested but the MIC50 of LVFX and CPFX were < or = 0.39 microgram/ml. Among anaerobes, Propionibacterium acnes was more susceptible than Peptostreptococcus species, and the MIC90 of beta-lactams and fluoroquinolones tested were < or = 0.78 microgram/ml. In conclusion, this study, performed on large number of strains, confirmed an excellent and wide spectrum antibacterial activity of LVFX compared with the fluoroquinolones and beta-lactams tested. And our results suggest that LVFX may be useful in the treatment of various bacterial infections. PMID:10746192

  7. In vitro anticancer activity, toxicity and structure-activity relationships of phyllostictine A, a natural oxazatricycloalkenone produced by the fungus Phyllosticta cirsii

    SciTech Connect

    Le Calve, Benjamin; Lallemand, Benjamin; Perrone, Carmen; Lenglet, Gaelle; Depauw, Sabine; Van Goietsenoven, Gwendoline; Bury, Marina; Vurro, Maurizio; Herphelin, Francoise; Andolfi, Anna; Zonno, Maria Chiara; Mathieu, Veronique; Dufrasne, Francois; Van Antwerpen, Pierre; Poumay, Yves

    2011-07-01

    The in vitro anticancer activity and toxicity of phyllostictine A, a novel oxazatricycloalkenone recently isolated from a plant-pathogenic fungus (Phyllosticta cirsii) was characterized in six normal and five cancer cell lines. Phyllostictine A displays in vitro growth-inhibitory activity both in normal and cancer cells without actual bioselectivity, while proliferating cells appear significantly more sensitive to phyllostictine A than non-proliferating ones. The main mechanism of action by which phyllostictine displays cytotoxic effects in cancer cells does not seem to relate to a direct activation of apoptosis. In the same manner, phyllostictine A seems not to bind or bond with DNA as part of its mechanism of action. In contrast, phyllostictine A strongly reacts with GSH, which is a bionucleophile. The experimental data from the present study are in favor of a bonding process between GSH and phyllostictine A to form a complex though Michael attack at C=C bond at the acrylamide-like system. Considering the data obtained, two new hemisynthesized phyllostictine A derivatives together with three other natural phyllostictines (B, C and D) were also tested in vitro in five cancer cell lines. Compared to phyllostictine A, the two derivatives displayed a higher, phyllostictines B and D a lower, and phyllostictine C an almost equal, growth-inhibitory activity, respectively. These results led us to propose preliminary conclusions in terms of the structure-activity relationship (SAR) analyses for the anticancer activity of phyllostictine A and its related compounds, at least in vitro.

  8. In vitro metabolism and bioavailability tests for endocrine active substances: What is needed next for regulatory purposes?

    EPA Science Inventory

    Legistation and prospective legislative proposals internationally (may) require that chemicals be tested for their ability to disrupt the hormonal systems of mammals. Chemicals found to test positive in vitro are considered to be endocrine active substances (EAS) and may be puta...

  9. In Vitro Assays for Assessment of Androgenic and Estrogenic Activity of Defined Mixtures and Complex Environmental Samples

    EPA Science Inventory

    Point sources of endocrine active compounds to aquatic environments such as waste water treatment plants, pulp and paper mills, and animal feeding operations invariably contain complex mixtures of chemicals. The current study investigates the use of targeted in vitro assays des...

  10. THE ROLE OF CCL4 BIOTRANSFORMATION IN THE ACTIVATION OF HEPATOCYTE PHOSPHOLIPASE C IN VIVO AND IN VITRO

    EPA Science Inventory

    The role of CC14 Biotransformation in the Activation of Hepatocyte Phospholipase C in ivo and in Vitro. Coleman, J. B., Condie, L.W. AND LAMB, R.G. (1988). Toxicol. Appl. Pharmacol. 95,208-219. Rats treated with a single 0.5 ml/kg dose (ip) of CCl4 exhibited a threefold increase ...

  11. Activation of Mouse Cumulus-Oocyte Complex Maturation In Vitro Through EGF-Like Activity of Versican.

    PubMed

    Dunning, Kylie R; Watson, Laura N; Zhang, Voueleng J; Brown, Hannah M; Kaczmarek, Adrian K; Robker, Rebecca L; Russell, Darryl L

    2015-05-01

    In vitro maturation of oocytes is suboptimal to in vivo maturation with altered gene expression and compromised oocyte quality. The large proteoglycan versican is abundant in mouse cumulus-oocyte complexes (COCs) matured in vivo but is absent in cultured COCs. Versican is also positively correlated with human oocyte quality. Versican contains an epidermal growth factor (EGF) motif, and based on EGF-like activities in other systems we hypothesized that versican acts as an EGF-like signaling factor during COC maturation. Here, we purified recombinant versican and compared its function with that of EGF during in vitro maturation (IVM). Versican significantly increased cumulus expansion and induced cumulus-specific genes Ptgs2, Tnfaip6, and Has2, which was blocked by EGF receptor antagonist. Microarray analysis revealed that versican has overlapping function with EGF; however, a subset of genes was uniquely altered following 6 h of IVM with either treatment. Following 6 h of IVM, both Areg and Ereg were significantly increased by both treatments, whereas Egln3, Nr4a1, Nr4a2, Nr4a3, and Adamts5 were significantly higher following versican treatment compared with EGF. In contrast, Sprr1a and Aqp3 were increased after 6 h of EGF but not versican treatment. To determine whether there were temporal differences, COCs were cultured with EGF or versican for 0-12 h. Versican-induced expression occurred later but remained elevated for longer compared with EGF for Ptgs2, Ereg, and Nr4a3. The unique expression profiles of Aqp3 and Nr4a3 during IVM were similarly regulated in vivo. These data indicate that versican has EGF-like effects on COC gene expression, but with distinct temporal characteristics. PMID:25810476

  12. Empirical modeling of an in vitro activity of polychlorinated biphenyl congeners and mixtures.

    PubMed Central

    Svendsgaard, D J; Ward, T R; Tilson, H A; Kodavanti, P R

    1997-01-01

    The goal of this research is to predict an in vitro activity of polychlorinated biphenyl (PCB) congeners and their mixtures and to describe the relationship between this activity and chemical structure. The test system used multiple PCB concentrations on each cell culture plate in a repeated measures design, which improved precision for comparing between concentration levels. A weighted regression that accounted for this experimental design feature was used in fitting a nonlinear dose-response exponential model to the PCB concentration-activity data from an in vitro test system in which 3H-phorbol ester binding was measured in cerebellar granule cells exposed to different PCB congeners to test for their effects on protein kinase C translocation. The model allowed for the minimum level to be less than control, a common slope, and the estimation of the log of the concentration that produces an activity 50% above the control activity (E50) for 36 congeners and 3 commercial mixtures. Next, a weighted logistic regression using a second order response model in the variables Clortho, Clpara, and Clmeta was used to relate the estimated log E50s to indicators of chemical structure. This model was preferred over models that might seem more mechanistically based because in internal validation, it attained a smaller PRESS statistic (the sum of squares between all observed and predicted observations) than other models. Evidently, this second order model makes more efficient use of parameters than other models considered. Plots of the predictions of the logistic second order response model versus log Kow confirm the usual pattern that congeners with intermediate levels of log Kow are the more active. The data of three commercial mixtures were included in this regression by assuming a common combination index (ratio of observed E50 to predicted E50, assuming dose addition). The logistic model suggests that congeners with one, two, or three chlorine substitutions at the ortho position are more active than other congeners. Also, congeners with log Kow between 5.2 and 6.6 are generally more active. The estimated combination index indicated that the joint action of PCB congeners in the three commercial mixtures was less than dose additive. The error sum of squares was significantly large, which may indicate a lack of fit of the logistic model. Empirical Bayes estimates (EBE) are weighted averages of model predictions and observations of E50s and can be better estimates than the fitted model when there is a lack of fit. The PRESS statistic for the EBE indicated larger prediction error than for the logistic model, but the EBE provided better estimates of commercial mixture E50s based on dose addition. This may indicate that the logistic model is not incorporating all the information in the single congener data needed to predict mixtures. Images Figure 1. Figure 2. Figure 2. Figure 2. Figure 2. Figure 2. Figure 2. Figure 3. Figure 4. Figure 4. Figure 4. Figure 4. Figure 4. Figure 4. Figure 5. Figure 5. Figure 5. Figure 5. Figure 5. Figure 5. PMID:9349838

  13. In vitro effect of recombinant interferon gamma in combination with LPS on amoebicidal activity of murine Kupffer cells.

    PubMed

    Ghadirian, E; Salimi, A

    1993-06-01

    The present study examines the role of liver macrophages (Kupffer cells), of C57BL/6 mice, as effector cells responsible for the killing of Entamoeba histolytica trophozoites in vitro. It was shown that unstimulated Kupffer cells were inefficient in the killing of E. histolytica trophozoites in vitro. Interferon gamma (IFN-gamma) alone was not able to activate Kupffer cells to amoebicidal state. However, Interferon gamma and lipopolysaccharide (LPS) acted synergistically in this phenomenon. It seems that the acquisition of amoebicidal activity is associated with the involvement of hydrogen peroxide, because the addition of catalase partially decreases the killing of this parasite by Kupffer cells. In addition, it appears that the amoebicidal activity of IFN-gamma-treated Kupffer cells is contact-dependent. Our results indicate that the immunologic production of IFN-gamma is important in the activation of Kupffer cells for controlling this parasite and that Kupffer cells are strong effector cells against the amoebae. PMID:8406558

  14. Plant proteolytic enzyme papain abrogates angiogenic activation of human umbilical vein endothelial cells (HUVEC) in vitro

    PubMed Central

    2013-01-01

    Background Vascular endothelial growth factor (VEGF) is a key regulator of physiologic and pathogenic angiogenesis in diseases such as cancer and diabetic retinopathy. It is known that cysteine proteases from plants, like bromelain and papain are capable to suppress inflammatory activation. Recent studies have demonstrated that they may interfere with angiogenesis related pathways as well. The aim of this study was to investigate the anti-angiogenic effects of papain on human umbilical vein endothelial cells (HUVEC) in vitro. Methods Cell viability after prolonged treatment with papain was investigated by life cell staining and lactate dehydrogenase release assay. Angiogenic activation was assessed by ELISA against phosphorylated proteins AKT, MEK1/2, ERK1/2, SAPK/JNK and p38-MAPK. Growth inhibition was determined by means of an MTT-assay and cell migration by means of a scratch assay. Capability to form a capillary network was investigated using a tube formation assay. Results Papain did not induce proteolysis or cell detachment of HUVEC in a concentration range between 0 and 25 ?g/mL. Four hours treatment with 10 ?g/mL papain resulted in a reduced susceptibility of endothelial cells to activation by VEGF as determined by phosphorylation levels of Akt, MEK1/2, SAPK/JNK. Papain exerted a distinct inhibitory effect on cell growth, cell migration and tube formation with inhibition of tube formation detectable at concentrations as low as 1 ?g/mL. Bromelain and ficin displayed similar effects with regard to cell growth and tube formation. Conclusion Papain showed a strong anti-angiogenic effect in VEGF activated HUVEC. This effect may be due to interference with AKT, MEK1/2 and SAPK/JNK phosphorylation. Two other plant derived cysteine proteases displayed similar inhibition of HUVEC cell growth and tube formation. These findings indicate that plant proteolytic enzymes may have potential as preventive and therapeutic agents against angiogenesis related human diseases. PMID:24053149

  15. In Vitro Antibacterial Activity of Several Plant Extracts and Oils against Some Gram-Negative Bacteria

    PubMed Central

    Al-Mariri, Ayman; Safi, Mazen

    2014-01-01

    Background: Medicinal plants are considered new resources for producing agents that could act as alternatives to antibiotics in the treatment of antibiotic-resistant bacteria. The aim of this study was to evaluate the antibacterial activity of 28 plant extracts and oils against four Gram-negative bacterial species. Methods: Experimental, in vitro, evaluation of the activities of 28 plant extracts and oils as well as some antibiotics against E. coli O157:H7, Yersinia enterocolitica O9, Proteus spp., and Klebsiella pneumoniae was performed. The activity against 15 isolates of each bacterium was determined by disc diffusion method at a concentration of 5%. Microdilution susceptibility assay was used in order to determine the minimal inhibitory concentrations (MICs) of the plant extracts, oils, and antibiotics. Results: Among the evaluated herbs, only Origanum syriacum L., Thymus syriacus Boiss., Syzygium aromaticum L., Juniperus foetidissima Wild, Allium sativum L., Myristica fragrans Houtt, and Cinnamomum zeylanicum L. essential oils and Laurus nobilis L. plant extract showed anti-bacterial activity. The MIC50 values of these products against the Gram-negative organisms varied from 1.5 (Proteus spp. and K. pneumoniae( and 6.25 µl/ml (Yersinia enterocolitica O9 ) to 12.5 µl/ml (E. coli O:157). Conclusion: Among the studied essential oils, O. syriacum L., T. syriacus Boiss., C. zeylanicum L., and S. aromaticum L. essential oils were the most effective. Moreover, Cephalosporin and Ciprofloxacin were the most effective antibiotics against almost all the studied bacteria. Therefore, O. syriacum L., T. syriacus Boiss., C. zeylanicum L., and S. aromaticum L. could act as bactericidal agents against Gram-negative bacteria. PMID:24453392

  16. Widely Used Pesticides with Previously Unknown Endocrine Activity Revealed as in Vitro Antiandrogens

    PubMed Central

    Orton, Frances; Rosivatz, Erika; Scholze, Martin; Kortenkamp, Andreas

    2011-01-01

    Background Evidence suggests that there is widespread decline in male reproductive health and that antiandrogenic pollutants may play a significant role. There is also a clear disparity between pesticide exposure and data on endocrine disruption, with most of the published literature focused on pesticides that are no longer registered for use in developed countries. Objective We used estimated human exposure data to select pesticides to test for antiandrogenic activity, focusing on highest use pesticides. Methods We used European databases to select 134 candidate pesticides based on highest exposure, followed by a filtering step according to known or predicted receptor-mediated antiandrogenic potency, based on a previously published quantitative structure–activity relationship (QSAR) model. In total, 37 pesticides were tested for in vitro androgen receptor (AR) antagonism. Of these, 14 were previously reported to be AR antagonists (“active”), 4 were predicted AR antagonists using the QSAR, 6 were predicted to not be AR antagonists (“inactive”), and 13 had unknown activity, which were “out of domain” and therefore could not be classified with the QSAR (“unknown”). Results All 14 pesticides with previous evidence of AR antagonism were confirmed as antiandrogenic in our assay, and 9 previously untested pesticides were identified as antiandrogenic (dimethomorph, fenhexamid, quinoxyfen, cyprodinil, ?-cyhalothrin, pyrimethanil, fludioxonil, azinphos-methyl, pirimiphos-methyl). In addition, we classified 7 compounds as androgenic. Conclusions Due to estimated antiandrogenic potency, current use, estimated exposure, and lack of previous data, we strongly recommend that dimethomorph, fludioxonil, fenhexamid, imazalil, ortho-phenylphenol, and pirimiphos-methyl be tested for antiandrogenic effects in vivo. The lack of human biomonitoring data for environmentally relevant pesticides presents a barrier to current risk assessment of pesticides on humans. PMID:21310686

  17. Flavonoid profiles of three Bupleurum species and in vitro hepatoprotective of activity Bupleurum flavum Forsk.

    PubMed Central

    Gevrenova, Reneta; Kondeva-Burdina, Magdalena; Denkov, Nikolay; Zheleva-Dimitrova, Dimitrina

    2015-01-01

    Background: Bupleurum L. (Aspiaceae) species are used as herbal remedy in Chinese traditional medicine. Objective: The aim was to investigate the flavonoids in three annual European Bupleurum species, including B. baldense, B. affine and B. flavum, and to test their antioxidant and possible hepatoprotective effects. Materials and Methods: Flavonoids from the methanol-aqueous extracts were quantified by solid-phase extraction-high-performance liquid chromatography. Bupleurum extracts (1–220 mg/ml) were tested for their antioxidant activity in DPPH and ABTS assays, as well as on isolated liver rat microsomes. In vitro hepatoprotective activity of B. flavum flavonoid (BFF) mixture and rutin, and narcissin, isolated from the same mixture, were evaluated on carbon tetrachloride (CCl4) and tert-butyl hydroperoxide (t-BuOOH) toxicity models in isolated rat hepatocytes. Results: Narcissin was the dominant flavonol glycoside in B. flavum being present at 24.21 ± 0.19 mg/g, whilst the highest content of rutin (28.63 ± 1.57 mg/g) was found in B. baldense. B. flavum possessed the strongest DPPH (IC50 22.12 ?g/ml) and ABTS (IC50 118.15 ?g/ml) activity. At a concentration 1 mg/ml of BFF (rutin 197.58 mg/g, narcissin 75.74 mg/g), a stronger antioxidant effect in microsomes was evidenced in comparison with silymarin, rutin and narcissin. The hepatoprotective effect of BFF significantly reduced the elevated levels of lactate dehydrogenase and malondialdehyde, and ameliorated glutathione, being most active in t-BuOOH-induced injury model when compared with CCl4 toxicity (P < 0.001). Conclusion: In BFF, synergism of rutin and narcissin could be responsible for stronger protection against mitochondrial induced oxidative stress. PMID:25709205

  18. In vitro antiplasmodial activity of PDDS-coated metal oxide nanoparticles against Plasmodium falciparum

    NASA Astrophysics Data System (ADS)

    Jacob Inbaneson, Samuel; Ravikumar, Sundaram

    2013-06-01

    Malaria is the most important parasitic disease, leading to annual death of about one million people and the Plasmodium falciparum develops resistant to well-established antimalarial drugs. The newest antiplasmodial drug from metal oxide nanoparticles helps in addressing this problem. Commercial nanoparticles such as Fe3O4, MgO, ZrO2, Al2O3 and CeO2 coated with PDDS and all the coated and non-coated nanoparticles were screened for antiplasmodial activity against P. falciparum. The Al2O3 nanoparticles (71.42 ± 0.49 ?g ml-1) showed minimum level of IC50 value and followed by MgO (72.33 ± 0.37 ?g ml-1) and Fe3O4 nanoparticles (77.23 ± 0.42 ?g ml-1). The PDDS-Fe3O4 showed minimum level of IC50 value (48.66 ± 0.45 ?g ml-1), followed by PDDS-MgO (60.28 ± 0.42 ?g ml-1) and PDDS-CeO2 (67.06 ± 0.61 ?g ml-1). The PDDS-coated metal oxide nanoparticles showed superior antiplasmodial activity than the non-PDDS-coated metal oxide nanoparticles. Statistical analysis reveals that, significant in vitro antiplasmodial activity ( P < 0.05) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes showed no morphological changes in erythrocytes by the nanoparticles after 48 h of incubation. It is concluded from the present study that, the PDDS-Fe3O4 showed good antiplasmodial activity and it might be used for the development of antiplasmodial drugs.

  19. Dose dependence of endotoxin-induced activation of the plasma contact system: an in vitro study.

    PubMed

    Roeise, O; Bouma, B N; Stadaas, J O; Aasen, A O

    1988-12-01

    The dose and time dependence of endotoxin-induced activation of the plasma contact system have been studied. Citrated pool plasma was incubated at 37 degrees C with endotoxin doses of 2.10(5), 2.10(6), 2.10(7), and 2.10(9) ng/l (lipopolysaccharide B, E. coli 026: B6, Difco Laboratories, Detroit, MI) for 24 hr. Samples for determination of components of the contact system were obtained prior to incubation and at 1, 2, 4, 6, 12, and 24 hr. Plasma kallikrein (KK) activity markedly increased at 12 hr in test plasma containing the highest dose of endotoxin (2.10(9) ng/l). Coincident with the elevated KK activity, reductions of both plasma prekallikrein (PKK) and functional kallikrein inhibition (KKI) were seen as assayed by chromogenic peptide substrate analyses. Also, functionally determined alpha 2-macroglobulin (alpha 2-M) and C1 inhibitor (C1INH) values were decreased, confirming the reduction of KKI values. Changes of Hageman factor (FXII), PKK, and high molecular weight kininogen (HMWK) values were also found at the same time point when assayed by immunochemical techniques. The same pattern of changes was seen in test plasma containing 2.10(7) and 2.10(6) ng/l of endotoxin. These changes, however, were less pronounced and not seen until 24 hr after beginning incubation. In control plasma and in plasma containing the lowest dose of endotoxin (2.10(5) ng/l), no changes were seen in any factors of the contact system. Our study shows that in vitro endotoxin-induced activation of the contact system is a slow process that is both time and dose dependent. PMID:2463883

  20. Stability and biological activity of wild blueberry (Vaccinium angustifolium) polyphenols during simulated in vitro gastrointestinal digestion.

    PubMed

    Correa-Betanzo, J; Allen-Vercoe, E; McDonald, J; Schroeter, K; Corredig, M; Paliyath, G

    2014-12-15

    Wild blueberries are rich in polyphenols and have several potential health benefits. Understanding the factors that affect the bioaccessibility and bioavailability of polyphenols is important for evaluating their biological significance and efficacy as functional food ingredients. Since the bioavailability of polyphenols such as anthocyanins is generally low, it has been proposed that metabolites resulting during colonic fermentation may be the components that exert health benefits. In this study, an in vitro gastrointestinal model comprising sequential chemostat fermentation steps that simulate digestive conditions in the stomach, small intestine and colon was used to investigate the breakdown of blueberry polyphenols. The catabolic products were isolated and biological effects tested using a normal human colonic epithelial cell line (CRL 1790) and a human colorectal cancer cell line (HT 29). The results showed a high stability of total polyphenols and anthocyanins during simulated gastric digestion step with approximately 93% and 99% of recovery, respectively. Intestinal digestion decreased polyphenol- and anthocyanin- contents by 49% and 15%, respectively, by comparison to the non-digested samples. During chemostat fermentation that simulates colonic digestion, the complex polyphenol mixture was degraded to a limited number of phenolic compounds such as syringic, cinnamic, caffeic, and protocatechuic acids. Only acetylated anthocyanins were detected in low amounts after chemostat fermentation. The catabolites showed lowered antioxidant activity and cell growth inhibition potential. Results suggest that colonic fermentation may alter the biological activity of blueberry polyphenols. PMID:25038707

  1. Optimization for ultrasound extraction of polysaccharides from mulberry fruits with antioxidant and hyperglycemic activity in vitro.

    PubMed

    Chen, Chun; You, Li-Jun; Abbasi, Arshad Mehmood; Fu, Xiong; Liu, Rui Hai

    2015-10-01

    Single-factor experiment and Box-Behnken design (BBD) were applied to optimize the ultrasound-assisted extraction of mulberry fruits polysaccharides (MFP). Under optimum conditions: ratio of water to raw material 40.25, extraction temperature 69°C, ultrasonic power 190W and extraction time 75 min, the MFP yield was 3.13% (±0.07%), in accordance to the predicted value of 3.04%. The mulberry fruits polysaccharides fractions was obtained by deproteinization (MFP-1), followed by decolorization and deionization (MFP-2). Carbohydrate content in MFP, MFP-1 and MFP-2 was 58.61% (±1.47%), 69.98% (±0.91%), 81.18% (±1.29%), as well as proteins was estimated 16.50% (±0.86%), 1.57% (±0.63%), 1.02% (±0.18%), respectively. The FT-IR indicated that MFP, MFP-1 and MFP-2 were acidic polysaccharides. The MFP-1 exhibited the strongest antioxidant activity, while MFP-2 showed the strongest hyperglycemic activity in vitro. This may be caused by their different compositions and physical properties in the different mulberry fruit polysaccharides fractions. PMID:26076608

  2. Antibacterial Activity, in Vitro Cytotoxicity, and Cell Cycle Arrest of Gemini Quaternary Ammonium Surfactants.

    PubMed

    Zhang, Shanshan; Ding, Shiping; Yu, Jing; Chen, Xuerui; Lei, Qunfang; Fang, Wenjun

    2015-11-10

    Twelve gemini quaternary ammonium surfactants have been employed to evaluate the antibacterial activity and in vitro cytotoxicity. The antibacterial effects of the gemini surfactants are performed on Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) with minimum inhibitory concentrations (MIC) ranging from 2.8 to 167.7 ?M. Scanning electron microscopy (SEM) analysis results show that these surfactants interact with the bacterial cell membrane, disrupt the integrity of the membrane, and consequently kill the bacteria. The data recorded on C6 glioma and HEK293 human kidney cell lines using an MTT assay exhibit low half inhibitory concentrations (IC50). The influences of the gemini surfactants on the cell morphology, the cell migration ability, and the cell cycle are observed through hematoxylin-eosin (HE) staining, cell wound healing assay, and flow cytometric analyses, respectively. Both the values of MIC and IC50 decrease against the growth of the alkyl chain length of the gemini surfactants with the same spacer group. In the case of surfactants 12-s-12, the MICs and IC50s are found to decrease slightly with the spacer chain length changing from 2 to 8 and again to increase at higher spacer length (s = 10-12). All of the gemini surfactants show great antibacterial activity and cytotoxicity, and they might exhibit potential applications in medical fields. PMID:26474336

  3. In vitro detection and quantification of botulinum neurotoxin type E activity in avian blood

    USGS Publications Warehouse

    Piazza, Timothy M.; Blehert, David S.; Dunning, F. Mark; Berlowski-Zier, Brenda M.; Zeytin, Fusun N.; Samuel, Michael D.; Tucker, Ward C.

    2011-01-01

    Botulinum neurotoxin serotype E (BoNT/E) outbreaks in the Great Lakes region cause large annual avian mortality events, with an estimated 17,000 bird deaths reported in 2007 alone. During an outbreak investigation, blood collected from bird carcasses is tested for the presence of BoNT/E using the mouse lethality assay. While sensitive, this method is labor-intensive and low throughput and can take up to 7 days to complete. We developed a rapid and sensitive in vitro assay, the BoTest Matrix E assay, that combines immunoprecipitation with high-affinity endopeptidase activity detection by Förster resonance energy transfer (FRET) to rapidly quantify BoNT/E activity in avian blood with detection limits comparable to those of the mouse lethality assay. On the basis of the analysis of archived blood samples (n = 87) collected from bird carcasses during avian mortality investigations, BoTest Matrix E detected picomolar quantities of BoNT/E following a 2-h incubation and femtomolar quantities of BoNT/E following extended incubation (24 h) with 100% diagnostic specificity and 91% diagnostic sensitivity.

  4. Characterization and in vitro antioxidant activity of Albizia stipulata Boiv. gum exudates.

    PubMed

    Thanzami, K; Malsawmtluangi, C; Lalhlenmawia, H; Seelan, T Veenus; Palanisamy, Selvamani; Kandasamy, Ruckmani; Pachuau, Lalduhsanga

    2015-09-01

    The objective of the present study is to characterize the physicochemical properties and to determine the in vitro antioxidant activity of Albizia stipulata Boiv. gum exudates collected from Northeast India. The total carbohydrate, uronic acid and protein contents, monosaccharide composition and the molecular weight distribution of the purified gum was determined. The powder flow property and preliminary compressibility test were performed on the dried gum exudates. Fourier transform infrared spectroscopy (FTIR) study was performed to analyze the functional groups present in the structure. Differential scanning calorimetry (DSC) and thermogravimetry (TGA/DTA) analyses were performed to study the thermal stability of the gum. The antioxidant properties of the gum were evaluated by determining 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl scavenging activities and reducing power. The total carbohydrate and protein contents of the gum were found to be 75.17±3.21% and 2.60±1.05% respectively. The viscosity of 2% aqueous solution of the gum exhibited non-Newtonian type of flow showing pH dependent swelling. Arabinose and galactose were found to be the main monosaccharides present in the gum exudates and the molecular weight distribution of the gum was also found to be polydispersed. Results from DPPH, hydroxyl scavenging and reducing power studies showed the gum possesses antioxidant properties. PMID:26118486

  5. In vitro and in vivo activities of trybizine hydrochloride against various pathogenic trypanosome species.

    PubMed

    Kaminsky, R; Brun, R

    1998-11-01

    Trybizine hydrochloride [O,O'-bis(4,6-diamino-1,2-dihydro-2, 2-tetramethylene-s-triazine-1-yl)-1,6-hexanediol dihydrochloride] was active in vitro against the sleeping sickness-causing agents Trypanosoma brucei subsp. rhodesiense and T. brucei subsp. gambiense; against a multidrug-resistant organism, T. brucei subsp. brucei; and against animal-pathogenic organisms Trypanosoma evansi, Trypanosoma equiperdum, and Trypanosoma congolense; but not against the intracellular parasites Trypanosoma cruzi and Leishmania donovani. Cytotoxic effects against mammalian cells were observed at approximately 10(6)-fold higher concentrations than those necessary to inhibit T. brucei subsp. rhodesiense. Trybizine hydrochloride was able to eliminate T. brucei subsp. rhodesiense and T. brucei subsp. gambiense in an acute rodent model with four intraperitoneal doses of 0.25 mg kg of body weight-1 or four doses of 1 mg kg-1, respectively, or with four oral doses of 20 mg kg-1. The compound expressed activity against suramin-resistant T. evansi strains in mice. However, these concentrations were not sufficient to cure mice infected with multidrug-resistant T. brucei subsp. brucei. A late-stage rodent model with central nervous system involvement could not be cured, indicating that trybizine may not pass the blood-brain barrier in sufficient quantities. PMID:9797216

  6. Comparative in vitro fungicidal activity of echinocandins against Candida albicans in peritoneal dialysis fluids.

    PubMed

    Tobudic, Selma; Forstner, Christina; Schranz, Heidelinde; Poeppl, Wolfgang; Vychytil, Andreas; Burgmann, Heinz

    2013-11-01

    The peritoneal dialysis (PD)-associated peritonitis caused by fungi is a relatively rare, but very serious disease. PD fluids (PDFs) affect inhibitory efficacy on the microorganisms' growth, which may compromise the affectivity of some antimicrobials. The purpose of this study was to investigate in vitro the fungicidal effectiveness of echinocandins in diverse PDFs. The fungicidal efficacy of caspofungin (CAS), anidulafungin (ANA), micafungin (MYC) against five clinical isolates of Candida albicans was studied in the different PDFs using time-kill curves. As control substance amphotericin B was used. Echinocandins showed slower and reduced killing of C. albicans in PDFs when compared with the time-kill curves in control bouillon. At concentration of 8 × minimal inhibitory concentration (MIC) the greatest reduction in the growth of C. albicans was seen by ANA in lactate-buffered Nutrineal PD4(®) with 1.1% amino acid (2.33 ± 0.52 log10 CFU ml(-1) ), and by CAS and MYC in lactate-buffered Dianeal PD4(®) with 1.36% glucose (2.36 ± 0.89 log10 CFU ml(-1) and 2.36 ± 0.99 log10 CFU ml(-1) respectively). Using high concentration of 128 × MIC echinocandins achieved fungicidal effect in all PDFs. PDFs may significantly impair the activities of echinocandins, but fungicidal activity of drugs can be achieved at high concentration of 128 × MIC. PMID:23551835

  7. In vitro antitumoral activity of compounds isolated from Artemisia gorgonum Webb.

    PubMed

    Martins, Alice; Mignon, Rukmini; Bastos, Marina; Batista, Daniela; Neng, Nuno R; Nogueira, José M F; Vizetto-Duarte, Catarina; Custódio, Luísa; Varela, Joăo; Rauter, Amélia P

    2014-09-01

    Artemisia gorgonum (Asteraceae) is an endemic plant to the Cape Verde islands and plays an important role in traditional medicine. The chloroform extract of the plant aerial parts afforded six sesquiterpene lactones, two methoxylated flavonoids, two lignans, and one tetracyclic triterpene, which were isolated by chromatographic methods and their structure established by physical and spectroscopic techniques. The cytotoxic activity of the three major constituents, namely, arborescin, artemetin, and sesamin, was evaluated on neuroblastoma (SH-SY5Y), hepatocarcinoma (HepG2), and nontumoral bone marrow stromal (S17) cell lines. The application of different concentrations of the compounds significantly decreased tumor cells viability at different extents, especially at the highest concentrations tested. Arborescin is the most promising compound as it was able to reduce tumoral cell viability with an IC50 significantly lower (229-233??M; p?activity in vitro, more pronounced on the cancer cell lines, confirming A.?gorgonum as a source of potential antitumoral molecules. PMID:24633846

  8. Antioxidant and anticancer activities of Chenopodium quinoa leaves extracts - in vitro study.

    PubMed

    Gawlik-Dziki, Urszula; ?wieca, Micha?; Su?kowski, Maciej; Dziki, Dariusz; Baraniak, Barbara; Czy?, Jaros?aw

    2013-07-01

    The nutraceutical potential of Chenopodium quinoa Leaves (ChL) was assessed through analyses of their phenolic content, elucidation of the effect of ChL phenolic compounds on cancer cell properties and estimation of their antioxidative activity, bioaccessibility and bioavailability in vitro. Considerable amounts of ferulic, sinapinic and gallic acids, kaempferol, isorhamnetin and rutin were observed in the chemical ChL extract and were linked with its inhibitory effect on prostate cancer cell proliferation, motility and cellular competence for gap junctional communication. Both extracts, chemical and obtained after simulated digestion, exerted an inhibitory effect on lipoxygenase activity, paralleled by their considerable chelating, antioxidative, antiradical and reducing power. These observations indicate that phenolic ChL compounds may exert a chemopreventive and anticarcinogenic effect on oxidative stress and ROS-dependent intracellular signaling via synergic effects. The relatively high potential bioaccessibility and bioavailability of the compounds probably responsible for these effects demonstrates the suitability of ChL for dietary supplementation. PMID:23537598

  9. In vitro activity of heather [Calluna vulgaris (L.) Hull] extracts on selected urinary tract pathogens

    PubMed Central

    Vu?i?, Dragana M.; Petkovi?, Miroslav R.; Rodi?-Grabovac, Branka B.; Stefanovi?, Olgica D.; Vasi?, Sava M.; ?omi?, Ljiljana R.

    2014-01-01

    Calluna vulgaris L. Hull (Ericaceae) has been used for treatment of urinary tract infections in traditional medicine. In this study we analyzed in vitro antibacterial activity of the plant extracts on different strains of Escherichia coli, Enterococcus faecalis and Proteus vulgaris, as well as the concentrations of total phenols and flavonoids in the extracts. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined. The concentrations of total phenols were examined by using Folin-Ciocalteu reagent and ranged between 67.55 to 142.46 mg GAE/g. The concentrations of flavonoids in extracts were determined using spectrophotometric method with aluminum chloride and the values ranged from 42.11 to 63.68 mg RUE/g. The aqueous extract of C. vulgaris showed a significant antibacterial activity. The values of MIC were in the range from 2.5 mg/ml to 20 mg/ml for this extract. Proteus vulgaris strains were found to be the most sensitive. The results obtained suggest that all tested extracts of C. vulgaris inhibit the growth of human pathogens, especially the aqueous extract. PMID:25428676

  10. Development of an Antioxidant Phytoextract of Lantana grisebachii with Lymphoprotective Activity against In Vitro Arsenic Toxicity

    PubMed Central

    Soria, Elio A.; Quiroga, Patricia L.; Albrecht, Claudia; Ramos Elizagaray, Sabina I.; Cantero, Juan J.; Bongiovanni, Guillermina A.

    2014-01-01

    Phytochemicals have been presumed to possess prophylactic and curative properties in several pathologies, such as arsenic- (As-) induced immunosuppression. Our aim was to discover a lymphoprotective extract from Lantana grisebachii Stuck. (Verbenaceae) (LG). We assessed its bioactivity and chemical composition using cell-based assays. Fractions produced from a hexane extract acutely induced nitrite formation in T-activated cell cultures (P < 0.0001). Water extraction released a fraction lacking nitrite inducing activity in both lymphocyte types. Aqueous LG was found to be safe in proliferated and proliferating cells. The infusion-derived extract presented better antioxidant capacity in proportion to phenolic amount in lymphocytes (infusive LG-1i at 100??g/mL), which protected them against in vitro As-induced lymphotoxicity (P < 0.0001). This infusive LG phytoextract contained 10.23 ± 0.43?mg/g of phenolics, with 58.46% being flavonoids. Among the phenolics, the only predominant compound was 0.723?mg of chlorogenic acid per gram of dry plant, in addition to 10 unknown minor compounds. A fatty acid profile was assessed. It contained one-third of saturated fatty acids, one-third of ?9, followed by ?6 (~24%) and ?3 (~4%), and scarce ?7. Summing up, L. grisebachii was a source of bioactive and lymphoprotective compounds, which could counteract As-toxicity. This supports its phytomedical use and research in order to reduce As-related dysfunctions. PMID:25002868

  11. In vitro and in vivo anti-allergic activity of soy sauce.

    PubMed

    Kobayashi, Makio; Matsushita, Hiroaki; Yoshida, Kazutoshi; Tsukiyama, Ryo-Ichi; Sugimura, Takashi; Yamamoto, Kenji

    2004-11-01

    Soy sauce (Shoyu) is a traditional fermented seasoning of Japan and available throughout the world. Polysaccharides were obtained from dialysate of Shoyu, and these Shoyu polysaccharides (SPS) were examined for anti-allergic activity in vitro and in vivo. The SPS originated from partially-degraded polysaccharides of soybeans by mold enzymatic hydrolyses, and Shoyu contained about 1% (w/v) SPS. First, the inhibitory effects of SPS on hyaluronidase, which is known to be related to inflammation and allergic responses, were as potent as those of an anti-allergic medicine, disodium cromoglycate. Second, SPS significantly inhibited the release of histamine from rat basophilic leukemia (RBL-2H3) cells, which had been induced by the antigen. Third, orally administered SPS had a significant suppressive effect on passive cutaneous anaphylaxis induced in the ears of mice. These results suggest that SPS may have anti-allergic activities, and soy sauce is a potentially promising seasoning for the treatment of allergic diseases through food. PMID:15492860

  12. In vitro antimycotic activity of a Williopsis saturnus killer protein against food spoilage yeasts.

    PubMed

    Goretti, Marta; Turchetti, Benedetta; Buratta, Morena; Branda, Eva; Corazzi, Lanfranco; Vaughan-Martini, Ann; Buzzini, Pietro

    2009-05-31

    The in vitro antimycotic activity of a purified killer protein (KT4561) secreted by a strain of Williopsis saturnus was tested against 310 yeast strains belonging to 21 food spoilage species of 14 genera (Candida, Debaryomyces, Dekkera, Hanseniaspora, Issatchenkia, Kazachstania, Kluyveromyces, Pichia, Rhodotorula, Saccharomyces, Schizosaccharomyces, Torulaspora, Yarrowia and Zygosaccharomyces). Minimum inhibitory concentration (MIC) determinations showed that over 65% of the target strains were susceptible to concentrations < or = 32 microg/ml of KT4561. Three conventional food-grade antimicrobial agents were used as controls: 41, 33 and 40% of the target strains were sensitive to < or = 512 mg/ml of ethyl 3-hydroxybenzoate (E214), potassium sorbate (E202) or potassium metabisulphite (E224), respectively. The susceptibility of food spoilage yeasts towards KT4561, E214, E202 and E224 was species- and strain-dependent. In most cases KT4561 exhibited MIC values several orders of magnitude lower (100 to 100,000 times) than those observed for E214, E202 and E224. With only a few exceptions, the activity of KT4561 was pH-, ethanol-, glucose- and NaCl-independent. The present study demonstrates the potential of this yeast killer protein as a novel and natural control agent against food spoilage yeasts. PMID:19269052

  13. In vitro antimicrobial activity of ZnO based glass-ceramics against pathogenic bacteria.

    PubMed

    Riaz, Madeeha; Zia, Rehana; Saleemi, Farhat; Ikram, Hafeez; Bashir, Farooq

    2015-12-01

    The antibacterial activity of ZnO (0-15.53 mol%) based SiO2-CaO-P2O5-Na2O-CaF2 bioactive glass-ceramics synthesized by controlled crystallisation were studied against eight micro-organisms using modified Kirby Bauer method. The antibacterial activity of the specimens was statistically evaluated using one-way analysis of variance and P < 0.05 was used as the level of significance. In vitro dissolution tests were performed in stimulated body fluid for 48 h at 37 °C for different time intervals to correlate the dissolution behaviour of test samples with antibacterial effects. The results illustrate that specimen BZn15.53 having the highest concentration of ZnO (15.53 mol %) demonstrated the strongest effect against Staph.aureus, S. epidermidis, B. subtilis and K. pneumonia. The effectiveness of BZn15.53 in inhibiting bacteria was due to accumulation of Zn(+2) ions around the surface of the bacteria cell release that caused the death of the cell, besides the presence of hydroxyapatite phase was also responsible for damaging the cell membrane of bacteria. PMID:26507201

  14. In Vitro Activity of Twenty Commercially Available, Plant-Derived Essential Oils against Selected Dermatophyte Species.

    PubMed

    Nardoni, Simona; Giovanelli, Silvia; Pistelli, Luisa; Mugnaini, Linda; Profili, Greta; Pisseri, Francesca; Mancianti, Francesca

    2015-08-01

    The in vitro activity of twenty chemically defined essential oils (EOs) obtained from Boswellia sacra, Citrus bergamia, C. limon, C. medica, Cinnamomum zeylanicum, Eucalyptus globulus, Foeniculum vulgare, Helichrysum italicum, Illicium verum, Litsea cubeba, Mentha spicata, Myrtus communis, Ocimum basilicum, Origanum majorana, O. vulgare, Pelargonium graveolens, Rosmarinus officinalis, Santalum album, Satureja montana, and Thymus serpyllum was assayed against clinical animal isolates of Microsporum canis, Trichophyton mentagrophytes, T. erinacei, T. terrestre and Microsporum gypseum, main causative agents of zoonotic and/or environmental dermatophytoses in humans. Single main components present in high amounts in such EOs were also tested. Different dermatophyte species showed remarkable differences in sensitivity. In general, more effective EOs were T. serpyllum (MIC range 0.025%-0.25%), O. vulgare (MIC range 0.025%-0.5%) and L. cubeba (MIC range 0.025%-1.5%). F. vulgare showed a moderate efficacy against geophilic species such as M gypseum and T terrestre. Among single main components tested, neral was the most active (MIC and MFC values 5 0.25%). The results of the present study seem to be promising for an in vivo use of some assayed EOs. PMID:26434145

  15. Antioxidant, Anti-Nephrolithe Activities and in Vitro Digestibility Studies of Three Different Cyanobacterial Pigment Extracts

    PubMed Central

    Paliwal, Chetan; Ghosh, Tonmoy; Bhayani, Khushbu; Maurya, Rahulkumar; Mishra, Sandhya

    2015-01-01

    Phycobiliprotein-containing water and carotenoid-containing methanolic extracts of three different cyanobacteria, Pseudanabaena sp., Spirulina sp. and Lyngbya sp., were studied for their DPPH scavenging, iso-bolographic studies, and anti-nephrolithe activities. The best EC50 values for DPPH scavenging were in Lyngbya water (LW, 18.78 ± 1.57 mg·mg?1 DPPH) and Lyngbya methanol (LM, 59.56 ± 37.38 mg·mg?1 DPPH) extracts. Iso-bolographic analysis revealed most of the combinations of extracts were antagonistic to each other, although LM—Spirulina methanol (SM) 1:1 had the highest synergistic rate of 86.65%. In vitro digestion studies showed that DPPH scavenging activity was considerably decreased in all extracts except for Pseudanabaena methanol (PM) and LM after the simulated digestion. All of the extracts were effective in reducing the calcium oxalate crystal size by nearly 60%–65% compared to negative control, while PM and Spirulina water (SW) extracts could inhibit both nucleation and aggregation of calcium oxalate by nearly 60%–80%. PMID:26308007

  16. In vitro activity of heather [Calluna vulgaris (L.) Hull] extracts on selected urinary tract pathogens.

    PubMed

    Vu?i?, Dragana M; Petkovi?, Miroslav R; Rodi?-Grabovac, Branka B; Stefanovi?, Olgica D; Vasi?, Sava M; Comi?, Ljiljana R

    2014-01-01

    Calluna vulgaris L. Hull (Ericaceae) has been used for treatment of urinary tract infections in traditional medicine. In this study we analyzed in vitro antibacterial activity of the plant extracts on different strains of Escherichia coli, Enterococcus faecalis and Proteus vulgaris, as well as the concentrations of total phenols and flavonoids in the extracts. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined. The concentrations of total phenols were examined by using Folin-Ciocalteu reagent and ranged between 67.55 to 142.46 mg GAE/g. The concentrations of flavonoids in extracts were determined using spectrophotometric method with aluminum chloride and the values ranged from 42.11 to 63.68 mg RUE/g. The aqueous extract of C. vulgaris showed a significant antibacterial activity. The values of MIC were in the range from 2.5 mg/ml to 20 mg/ml for this extract. Proteus vulgaris strains were found to be the most sensitive. The results obtained suggest that all tested extracts of C. vulgaris inhibit the growth of human pathogens, especially the aqueous extract. PMID:25428676

  17. In Vitro and In Vivo Antibacterial Activity of Punica granatum Peel Ethanol Extract against Salmonella

    PubMed Central

    Choi, Jang-Gi; Kang, Ok-Hwa; Lee, Young-Seob; Chae, Hee-Sung; Oh, You-Chang; Brice, Obiang-Obounou; Kim, Min-San; Sohn, Dong-Hwan; Kim, Hun-Soo; Park, Hyun; Shin, Dong-Won; Rho, Jung-Rae; Kwon, Dong-Yeul

    2011-01-01

    Punica granatum is commonly used in Korea as a traditional medicine for the treatment of pathogenic bacteria. In this study, we investigated the in vitro and in vivo antimicrobial activity of P. granatum peel EtOH extract (PGPE) against 16 strains of Salmonella. The minimal inhibitory concentrations of PGPE were in the range of 62.5–1000?x03BCg?mL?1. In addition, the in vivo antibacterial activity of the PGPE extract was examined in a S. typhimurium infection mouse model. Mice were initially infected with S. typhimurium and then with PGPE. The extract was found to have significant effects on mortality and the numbers of viable S. typhimurium recovered from feces. Although clinical signs and histological damage were rarely observed in the treated mice, the untreated controls showed signs of lethargy and histological damage in the liver and spleen. Taken together, the results of this study indicate that PGPE has the potential to provide an effective treatment for salmonellosis. PMID:19687188

  18. Preparation, characterisation and study of in vitro biologically active azamacrocyclic Cu(II) dicarboxylate complexes

    NASA Astrophysics Data System (ADS)

    Antonijevi?-Nikoli?, Mirjana; Anti?-Stankovi?, Jelena; Tanaskovi?, Sladjana B.; Korabik, Maria J.; Gojgi?-Cvijovi?, Gordana; Vu?kovi?, Gordana

    2013-12-01

    New cationic Cu(II) complexes with N, N?, N?, N??-tetrakis(2-pyridylmethyl)-1,4,8,11-tetraazacyclotetradecane (tpmc) and aliphatic dicarboxylic acids: pentanedioic (glutaric acid = glutH2), hexanedioic acid (adipic acid = adipH2) and decanedioic acid (sebacic acid = sebH2) of general formula [Cu4(L)(tpmc)2](ClO4)6·xH2O, L = glut, x = 2; L = adip, x = 7; L = seb, x = 6 were isolated. Their composition and charges are proposed based on elemental analyses and molar conductivity measurements. By the comparison of their UV-Vis, reflectance, FTIR and EPR spectral data, CV and SQUID magnetic measurements, with those for the complex with butanedioic acid (succinic acid = succH2) of known molecular structure and analysis of LC/MS spectra, geometry with two [Cu2tpmc]4+ units bridged by dicarboxylate dianion engaging all oxygens is proposed. Within units, Cu(II) ions are also bridged with N portion of cyclam ring. All four complexes were screened to in vitro antimicrobial and cytotoxic activity along with free primary and secondary ligands, Cu(II) salt and solvent controls. Detected antibacterial and cytotoxic activity for the complexes was enhanced in most cases than the corresponding controls.

  19. A light-activated metal complex targets both DNA and RNA in a fluorescent in vitro transcription and translation assay.

    PubMed

    Heidary, David K; Glazer, Edith C

    2014-03-01

    A coupled in vitro transcription and translation (IVTT) assay that uses GFP as a fluorescent reporter allowed the potency of a light-activated cytotoxic ruthenium agent to be quantified. The compound inhibits the function of both DNA and mRNA only upon light activation. The IVTT functional assay provides estimates of potency that are consistent with cellular cytotoxicity values, in marked contrast to the values obtained from traditional DNA-damage assays. PMID:24482049

  20. Cytotoxic activity of Thai medicinal plants against human cholangiocarcinoma, laryngeal and hepatocarcinoma cells in vitro

    PubMed Central

    2010-01-01

    Background Cholangiocarcinoma is a serious public health in Thailand with increasing incidence and mortality rates. The present study aimed to investigate cytotoxic activities of crude ethanol extracts of a total of 28 plants and 5 recipes used in Thai folklore medicine against human cholangiocarcinoma (CL-6), human laryngeal (Hep-2), and human hepatocarcinoma (HepG2) cell lines in vitro. Methods Cytotoxic activity of the plant extracts against the cancerous cell lines compared with normal cell line (renal epithelial cell: HRE) were assessed using MTT assay. 5-fluorouracil was used as a positive control. The IC50 (concentration that inhibits cell growth by 50%) and the selectivity index (SI) were calculated. Results The extracts from seven plant species (Atractylodes lancea, Kaempferia galangal, Zingiber officinal, Piper chaba, Mesua ferrea, Ligusticum sinense, Mimusops elengi) and one folklore recipe (Pra-Sa-Prao-Yhai) exhibited promising activity against the cholangiocarcinoma CL-6 cell line with survival of less than 50% at the concentration of 50 ?g/ml. Among these, the extracts from the five plants and one recipe (Atractylodes lancea, Kaempferia galangal, Zingiber officinal, Piper chaba, Mesua ferrea, and Pra-Sa-Prao-Yhai recipe) showed potent cytotoxic activity with mean IC50 values of 24.09, 37.36, 34.26, 40.74, 48.23 and 44.12 ?g/ml, respectively. All possessed high activity against Hep-2 cell with mean IC50 ranging from 18.93 to 32.40 ?g/ml. In contrast, activity against the hepatoma cell HepG2 varied markedly; mean IC50 ranged from 9.67 to 115.47 ?g/ml. The only promising extract was from Zingiber officinal (IC50 = 9.67 ?g/ml). The sensitivity of all the four cells to 5-FU also varied according to cell types, particularly with CL-6 cell (IC50 = 757 micromolar). The extract from Atractylodes lancea appears to be both the most potent and most selective against cholangiocarcinoma (IC50 = 24.09 ?g/ml, SI = 8.6). Conclusions The ethanolic extracts from five plants and one folklore recipe showed potent cytotoxic activity against CL-6 cell. Sensitivity to other cancerous cell lines varied according to cell types and the hepatocarcinoma cell line. HepG2 appears to be the most resistant to the tested extracts. PMID:20920194

  1. Evaluation of Ceftaroline Activity versus Ceftriaxone against Clinical Isolates of Streptococcus pneumoniae with Various Susceptibilities to Cephalosporins in an In Vitro Pharmacokinetic/Pharmacodynamic Model

    E-print Network

    Steed, Molly E.; Vidaillac, Celine; Winterfield, Patricia; Biek, Donald; Rybak, Michael J.

    2012-05-01

    Drug resistance in Streptococcus pneumoniae, a frequent pathogen in community-acquired pneumonia, is increasing. Ceftaroline (active metabolite of ceftaroline fosamil) is a broad-spectrum intravenous cephalosporin with activity in vitro against drug...

  2. Studies of the in vitro cytotoxic, antioxidant, lipase inhibitory and antimicrobial activities of selected Thai medicinal plants

    PubMed Central

    2012-01-01

    Background Traditional folk medicinal plants have recently become popular and are widely used for primary health care. Since Thailand has a great diversity of indigenous (medicinal) plant species, this research investigated 52 traditionally used species of Thai medicinal plants for their in vitro cytotoxic, antioxidant, lipase inhibitory and antimicrobial activities. Methods The 55 dried samples, derived from the medicinally used parts of the 52 plant species were sequentially extracted by hexane, dichloromethane, ethanol and water. These 220 extracts were then screened for in vitro (i) cytotoxicity against four cell lines, derived from human lung (A549), breast (MDA-MB-231), cervical (KB3-1) and colon (SW480) cancers, using the MTT cytotoxicity assay; (ii) antioxidant activity, analyzed by measuring the scavenging activity of DPPH radicals; (iii) lipase inhibitory activity, determined from the hydrolytic reaction of p-nitrophenyllaurate with pancreatic lipase; and (iv) antimicrobial activity against three Gram-positive and two Gram-negative bacteria species plus one strain of yeast using the disc-diffusion method and determination of the minimum inhibitory concentration by the broth micro-dilution assay. Results The crude dichloromethane and/or ethanol extracts from four plant species showed an effective in vitro cytotoxic activity against the human cancer cell lines that was broadly similar to that of the specific chemotherapy drugs (etoposide, doxorubicin, vinblastine and oxaliplatin). In particular, this is the first report of the strong in vitro cytotoxic activity of Bauhinia strychnifolia vines. The tested tissue parts of only six plant species (Allium sativum, Cocoloba uvifera, Dolichandrone spathacea, Lumnitzera littorea, Sonneratia alba and Sonneratia caseolaris) showed promising potential antioxidant activity, whereas lipase inhibitory activity was only found in the ethanol extract from Coscinum fenestratum and this was weak at 17-fold lower than Orlistat, a known lipase inhibitor. The highest antimicrobial activity was observed in the extracts from S. alba and S. caseolaris against Pseudomonas aeruginosa and Candida albicans, respectively. Conclusion The Thai medicinal plant B. strychnifolia is first reported to exert strong in vitro cytotoxic activities against human cancer cell lines and warrants further enrichment and characterization. The broad spectrum of the biological activities from the studied plant extracts can be applied as the guideline for the selection of Thai medicinal plant species for further pharmacological and phytochemical investigations. PMID:23145786

  3. In vitro antifungal activity of antipsychotic drugs and their combinations with conventional antifungals against Scedosporium and Pseudallescheria isolates.

    PubMed

    Homa, Mónika; Galgóczy, László; Tóth, Eszter; Tóth, Liliána; Papp, Tamás; Chandrasekaran, Muthusamy; Kadaikunnan, Shine; Alharbi, Naiyf S; Vágvölgyi, Csaba

    2015-11-01

    In the present study, in vitro antifungal activities of five antipsychotic drugs (i.e., chlorpromazine hydrochloride, CPZ; trifluoperazine hydrochloride, TPZ; amantadine hydrochloride; R-(-)-deprenyl hydrochloride, and valproic acid sodium salt) and five conventional antifungal drugs (i.e., amphotericin B, AMB; caspofungin, CSP; itraconazole; terbinafine, TRB and voriconazole, VRC) were investigated in broth microdilution tests against four clinical and five environmental Scedosporium and Pseudallescheria isolates. When used alone, phenothiazines CPZ and TPZ exerted remarkable antifungal effects. Thus, their in vitro combinations with AMB, CSP, VRC, and TRB were also examined against the clinical isolates. In combination with antifungal agents, CPZ was able to act synergistically with AMB and TRB in cases of one and two isolates, respectively. In all other cases, indifferent interactions were revealed. Antagonism was not observed between the tested agents. These combinations may establish a more effective and less toxic therapy after further in vitro and in vivo studies for Scedosporium and Pseudallescheria infections. PMID:26316212

  4. Activity of oritavancin and comparators in vitro against standard and high inocula of Staphylococcus aureus.

    PubMed

    Arhin, Francis F; Sarmiento, Ingrid; Parr, Thomas R; Moeck, Gregory

    2012-02-01

    In this study, the impact of inoculum density on the growth inhibitory and killing activities of oritavancin and comparators (vancomycin, daptomycin and linezolid) in vitro against four Staphylococcus aureus strains at clinically relevant drug concentrations was studied. Broth microdilution and time-kill assays were performed using a standard inoculum [ca. 10(5)colony-forming units (CFU)/mL as per Clinical and Laboratory Standards Institute (CLSI) guidelines] and a high inoculum (ca. 10(7)CFU/mL). Whereas minimal inhibitory concentrations (MICs) of comparators were 2-8-fold higher when tested at high inoculum, oritavancin MICs were 16-fold higher for all strains at the high inoculum relative to the standard inoculum. However, in time-kill assays, when tested at its fC(min) [trough concentration of free (non-protein-bound) drug] and fC(max) (peak concentration of non-protein-bound drug), oritavancin retained its bactericidal activity against a vancomycin-susceptible, meticillin-susceptible S. aureus (VS-MSSA) strain and a vancomycin-susceptible, meticillin-resistant S. aureus (VS-MRSA) strain both at standard and high inocula. At its fC(max), oritavancin was bactericidal at standard inoculum but not at high inoculum against two vancomycin-intermediate S. aureus (VISA) strains. Against both VISA strains at standard inoculum, oritavancin at its fC(min) reduced cell density by between 2 and 3 log (bacteriostatic), predicting that it will retain activity against certain VISA infections. However, oritavancin had no substantial growth inhibitory effect against either VISA strain at high inoculum, suggesting that in rare VISA infections with an anticipated high bacterial burden such as endocarditis, alternative oritavancin dosing strategies, including combinations with other agents, may be explored. PMID:22070858

  5. Contributions of intrinsic and synaptic activities to the generation of neuronal discharges in in vitro hippocampus

    PubMed Central

    Cohen, Ivan; Miles, Richard

    2000-01-01

    Extracellular and intracellular records were made from guinea-pig hippocampal slices to examine the contributions of intrinsic cellular properties and synaptic events to the generation of neuronal activity. Extracellular signals were filtered to pass action potentials, which could be detected within a distance of about 80 ?m from a discharging cell.Spontaneous action potentials were invariably detected in records from the stratum pyramidale of CA3 region. Blocking excitatory synaptic transmission with NBQX and APV reduced their frequency by 23 ± 35 %. Suppressing synaptic inhibition, while excitation was already blocked, increased the rate of spike discharge to 177 ± 71 % of its control value.Most action potentials recorded intracellularly from CA3 pyramidal cells were initiated in the absence of a detectable synaptic event. In contrast, most action potentials generated by inhibitory cells located close to stratum pyramidale were preceded by an EPSP.In 31 simultaneous recordings, intracellular pyramidal cell action potentials appeared consistently to initiate extracellular spikes with a mean latency of 2·2 ± 1·0 ms. Single inhibitory cell action potentials could initiate a reduction in the frequency of extracellular spikes of duration 10–30 ms.Some identified extracellular spikes (n = 9) consistently preceded intracellularly recorded IPSPs. IPSPs were initiated monosynaptically with latencies of 0·9–1·5 ms. In reciprocal interactions, single pyramidal cell action potentials could trigger the discharge of an identified unit that in turn appeared to initiate an IPSP in the same pyramidal cell.These data suggest that intrinsic cellular mechanisms underly much of the spontaneous activity of pyramidal cells of the CA3 region of the hippocampus in vitro. Both synaptic inhibition and a strong excitation of inhibitory cells by pyramidal cells act to reduce population activity. PMID:10766928

  6. Free radical scavenging activity in in vitro-derived tissues of Eruca sativa.

    PubMed

    Abbasi, Bilal Haider; Ali, Jawad; Ali, Mohammad; Zia, Muhammad; Bokhari, Saleem A; Khan, Mubarak Ali

    2016-01-01

    Feasible regeneration protocol for economically important plant Eruca sativa was established and 1, 1-diphenyl-2-picrylhydrazyl scavenging activity of regenerated tissues was evaluated and compared with plant material collected from the wild. Leaf portions inoculated onto Murashige and Skoog (MS) medium responded to all plant growth regulators exploited. Optimum callus production was achieved on a combination of 2.0 mg l(-1) 6-benzyladenine (BA) + 1.0 mg l(-1) ?-naphthalene acetic acid (NAA) and the lowest response was recorded for 0.5 mg l(-1) gibberellic acid (GA3) + 1.0 mg l(-1) NAA. The callus was subcultured on similar composition/concentrations of plant growth regulators after 4 weeks of culture time. A 5.0 mg l(-1) 6-BA + 1.0 mg l(-1) NAA produced optimum percentage shoot organogenesis after 4 weeks of subculturing. However, optimum number of shoots per explant was recorded for moderate concentrations (1.0 and 2.0 mg l(-1)) of kinetin. Incorporation of NAA into MS medium-containing GA3 also produced a feasible number of shoots/explant. Similar mean shoot length was recorded for 2.0 mg l(-1) kinetin + 1.0 mg l(-1) NAA and optimum concentrations (2.0, 5.0, and 10.0 mg l(-1)) of GA3 + 1.0 mg l(-1) NAA. In vitro generated shoots were shifted to MS medium augmented with indole acetic acid (IAA) for rooting after 4 weeks of subculturing. Moderate concentrations (5.0 mg l(-1)) of IAA produced feasible rooting. Investigation of radical scavenging activity showed that callus possesses higher levels of radical scavengers than other plant tissues tested. Phenolics and glucosides are reported to be active components of Eruca sativa phytochemistry. PMID:24021434

  7. Glyphosate affects the spontaneous motoric activity of intestine at very low doses - in vitro study.

    PubMed

    Ch?opecka, Magdalena; Mendel, Marta; Dziekan, Natalia; Karlik, Wojciech

    2014-07-01

    Glyphosate is an active substance of the most popular herbicides worldwide. Its common use results from the belief that it affects exclusively plants. However, studies on glyphosate and its trade formulations reveal that it causes numerous morphological, physiological and biochemical disturbances in cells and organisms of animals, including mammals. Due to the fact that shortly after oral exposure glyphosate is detected in the highest amount in small intestine, the aim of this study was to evaluate the effect of this compound on the spontaneous motoric activity of intestine under in vitro conditions. The experiments were conducted on rat jejunum strips under isotonic conditions. The strips were incubated in buffered (pH 7.35) and non-buffered (pH 5.2) glyphosate solutions ranged from 0.003 to 1.7 g/L. The results indicate that glyphosate applied in buffered solution affects significantly the spontaneous motoric activity of rat isolated jejunum strips. The muscle response is biphasic (miorelaxation accompanied by contraction). The contraction is observed already at a dose of 0.003 g/L and the first significant biphasic reaction at a dose of 0.014 g/L. The incubation of jejunum strips with glyphosate in non-buffered solution (pH 5.2) results in a different reaction. The smooth muscle undergoes only persistent relaxation, which is stronger than the response to glyphosate solution in pH 7.35. Motility disturbances are also observed after glyphosate removal from the incubation solution. The gathered data suggests that glyphosate impairs gastrointestinal strips' motility at concentration that are noticed in human exposed to non-toxic doses of glyphosate. PMID:25052523

  8. Short communication: an in vitro assessment of the antibacterial activity of plant-derived oils.

    PubMed

    Mullen, K A E; Lee, A R; Lyman, R L; Mason, S E; Washburn, S P; Anderson, K L

    2014-09-01

    Nonantibiotic treatments for mastitis are needed in organic dairy herds. Plant-derived oils may be useful but efficacy and potential mechanisms of action of such oils in mastitis therapy have not been well documented. The objective of the current study was to evaluate the antibacterial activity of the plant-derived oil components of Phyto-Mast (Bovinity Health LLC, Narvon, PA), an herbal intramammary product, against 3 mastitis-causing pathogens: Staphylococcus aureus, Staphylococcus chromogenes, and Streptococcus uberis. Plant-derived oils evaluated were Thymus vulgaris (thyme), Gaultheria procumbens (wintergreen), Glycyrrhiza uralensis (Chinese licorice), Angelica sinensis, and Angelica dahurica. Broth dilution testing according to standard protocol was performed using ultrapasteurized whole milk instead of broth. Controls included milk only (negative control), milk + bacteria (positive control), and milk + bacteria + penicillin-streptomycin (antibiotic control, at 1 and 5% concentrations). Essential oil of thyme was tested by itself and not in combination with other oils because of its known antibacterial activity. The other plant-derived oils were tested alone and in combination for a total of 15 treatments, each replicated 3 times and tested at 0.5, 1, 2, and 4% to simulate concentrations potentially achievable in the milk within the pre-dry-off udder quarter. Thyme oil at concentrations ?2% completely inhibited bacterial growth in all replications. Other plant-derived oils tested alone or in various combinations were not consistently antibacterial and did not show typical dose-response effects. Only thyme essential oil had consistent antibacterial activity against the 3 mastitis-causing organisms tested in vitro. Further evaluation of physiological effects of thyme oil in various preparations on mammary tissue is recommended to determine potential suitability for mastitis therapy. PMID:25022682

  9. The In vitro anti-acne activity of two unani drugs

    PubMed Central

    Chaudhary, Shahid Shah; Tariq, Mohd.; Zaman, Roohi; Imtiyaz, Shaikh

    2013-01-01

    Background: Acne is the most common disorder treated by dermatologists. As many as 80-90% of all adolescents have some type of acne and 30% of them require medical treatment. It is an inflammatory disease of the pilosebaceous unit characterized by the formation of open and closed comedones, papules, pustules, nodules, and cysts. Aims: The present study was conducted to investigate the in vitro anti-acne activity of two Unani single drugs Darchini (Cinnamomum zeylanicum Bl.) and Tukhm Khashkhash (Papaver somniferum L. seeds). Materials and Methods: The antibacterial activity of aqueous, ethanolic and hydroalcoholic extracts of both drugs were investigated against two acne causing bacteria, i.e., Propionibacterium acne and Staphylococcus epidermidis using well diffusion method. Results: The result showed that both drugs were active against the two bacteria. Against P. acne aqueous and ethanolic extract of Darchini and Tukhm Khashkhash showed the zone of inhibition of 18 ± 1.02 mm and 18 ± 1.6 mm and 13 ± 1.04 mm and 14 ± 1.8 mm, respectively. Against S. epidermidis aqueous, hydroalcoholic and ethanolic extracts of Darchini showed 22 ± 1.7 mm, 22 ± 1.2 mm and 15 ± 1.8 mm zone of inhibition respectively. Hydroalcoholic and ethanolic extracts of Tukhm Khashkhash showed 15 ± 1.09 mm and 13 ± 1.6 mm zone of inhibition respectively. Conclusion: This suggests that C. zeylanicum and P. somniferum have potential against acne causing bacteria and hence they can be used in topical anti-acne preparations and may address the antibiotic resistance of the bacteria. PMID:25161328

  10. 2-Amino-thiophene derivatives present antileishmanial activity mediated by apoptosis and immunomodulation in vitro.

    PubMed

    Rodrigues, Klinger Antonio da Franca; Dias, Cínthia Nóbrega de Sousa; Néris, Patrícia Lima do Nascimento; Rocha, Juliana da Câmara; Scotti, Marcus Tullius; Scotti, Luciana; Mascarenhas, Sandra Rodrigues; Veras, Robson Cavalcante; Medeiros, Isac Almeida de; Keesen, Tatjana de Souza Lima; Oliveira, Tiago Bento de; Lima, Maria do Carmo Alves de; Balliano, Tatiane Luciano; Aquino, Thiago Mendonça de; Moura, Ricardo Olímpio de; Mendonça Junior, Francisco Jaime Bezerra; Oliveira, Márcia Rosa de

    2015-12-01

    This study evaluated the effects of 2-amino-thiophene derivatives on the promastigote and amastigote forms of Leishmania (Leishmania) amazonensis and their possible mechanisms of action. Initially, we evaluated the antileishmanial activity of ten 2-amino-thiophene derivatives on promastigote and axenic amastigote forms of Leishmania amazonensis and their cytotoxicity against murine macrophages and human red blood cells. Three promising compounds were selected for studies of the cell death process using flow cytometry analysis and a DNA fragmentation assay. The effects of the compounds were assessed on intramacrophagic amastigotes, and the modulation of cytokine and NO production was investigated. All thiophene derivatives showed antileishmanial activity against promastigotes and axenic amastigotes with less toxicity for murine macrophages and human red blood cells. The best values were obtained for compounds containing a lateral indole ring. Docking studies suggested that these compounds played an important role in inhibiting trypanothione reductase (TryR) activity. The selected compounds SB-200, SB-44, and SB-83 induced apoptosis in promastigotes involving phosphatidylserine externalization and DNA fragmentation in a pattern similar to that observed for the positive control. Additionally, SB-200, SB-44, and SB-83 significantly reduced the infection index of macrophages by the parasites; for compounds SB-200 and SB-83 this reduction was associated with increased TNF-?, IL-12, and NO levels. This study demonstrated the effective and selective action of 2-amino-thiophene derivatives against L. amazonensis, resulting in apoptosis-like cell death and immunomodulation in vitro. The results suggest that they are promising compounds for the development of new leishmanicidal drugs. PMID:26513640

  11. In Vitro Antiplasmodial Activities and Synergistic Combinations of Differential Solvent Extracts of the Polyherbal Product, Nefang

    PubMed Central

    Arrey Tarkang, Protus; Franzoi, Kathrin Diehl; Lee, Eunyoung; Freitas-Junior, Lucio; Liuzzi, Michel; Nolé, Tsabang; Ayong, Lawrence S.; Agbor, Gabriel A.; Okalebo, Faith A.; Guantai, Anastasia N.

    2014-01-01

    Nefang, a polyherbal product composed of Mangifera indica (bark and leaf), Psidium guajava, Carica papaya, Cymbopogon citratus, Citrus sinensis, and Ocimum gratissimum (leaves), is a potential therapy against P. falciparum malaria. In vitro antiplasmodial activities of its constituent solvent extracts were analyzed on CQ-sensitive (3D7) and multidrug resistant (Dd2) P. falciparum strains. The interactions involving the differential solvent extracts were further analyzed using a variable potency ratio drug combination approach. Effective concentration 50 (EC50) values were determined by nonlinear regression curve-fitting of the dose-response data and used in calculating the fractional inhibitory concentration 50 (FIC50) and combination indices (CI) for each pair. The derived EC50 values (3D7/Dd2, ?g/mL) are Nefang-96.96/55.08, MiB-65.33/34.58, MiL-82.56/40.04, Pg-47.02/25.79, Cp-1188/317.5, Cc-723.3/141, Cs-184.4/105.1, and Og-778.5/118.9. Synergism was obtained with MiB/Pg (CI = 0.351), MiL/Pg (0.358), MiB/Cs (0.366), MiL/Cs (0.482), Pg/Cs (0.483), and Cs/Og (0.414) when analyzed at equipotency ratios. Cytotoxicity testing of Nefang and the solvent extracts on two human cell lines (Hep G2 and U2OS) revealed no significant toxicity relative to their antiplasmodial activities (SI > 20). Taken together, our data confirm the antimalarial activities of Nefang and its constituent plant extracts and identified extract pairs with promising synergistic interactions for exploitation towards a rational phytotherapeutic and evidence-based antimalarial drug discovery. PMID:24877138

  12. In vitro activity of nitazoxanide and related compounds against isolates of Giardia intestinalis, Entamoeba histolytica and Trichomonas vaginalis.

    PubMed

    Adagu, I Sullayman; Nolder, Deborah; Warhurst, David C; Rossignol, Jean-François

    2002-01-01

    The activities of the N-(nitrothiazolyl) salicylamide nitazoxanide and its metabolite tizoxanide were compared with metronidazole in vitro in microplates against six axenic isolates of Giardia intestinalis. Tizoxanide was eight times more active than metronidazole against metronidazole-susceptible isolates and twice as active against a resistant isolate. In 10 axenic isolates of Entamoeba histolytica, while tizoxanide was almost twice as active as metronidazole against more susceptible isolates, it was more than twice as active against less susceptible isolates. Fourteen metronidazole-susceptible isolates of Trichomonas vaginalis were 1.5 times more susceptible to tizoxanide, which was nearly five times as active against resistant isolates. Two highly metronidazole-resistant isolates retained complete susceptibility to tizoxanide, and one moderately resistant isolate showed reduced susceptibility. In all three organisms, nitazoxanide results paralleled those of tizoxanide. Analogues lacking the reducible nitro-group had similar low activities against susceptible G. intestinalis, E. histolytica and T. vaginalis, indicating that nitro-reduction and free radical production was a probable mode of action. Nitazoxanide and its metabolite tizoxanide are more active in vitro than metronidazole against G. intestinalis, E. histolytica and T. vaginalis. Although, like metronidazole, they depend on the presence of a nitro-group for activity, they retain some activity against metronidazole-resistant strains, particularly of T. vaginalis. The results suggest that resistance mechanisms for metronidazole can be bypassed by nitazoxanide and tizoxanide. PMID:11751773

  13. Vaticaffinol, a resveratrol tetramer, exerts more preferable immunosuppressive activity than its precursor in vitro and in vivo through multiple aspects against activated T lymphocytes

    SciTech Connect

    Feng, Li-Li; Wu, Xue-Feng; Liu, Hai-Liang; Guo, Wen-Jie; Luo, Qiong; Tao, Fei-Fei; Ge, Hui-Ming; Shen, Yan; Tan, Ren-Xiang; Xu, Qiang Sun, Yang

    2013-03-01

    In the present study, we aimed to investigate the immunosuppressive activity of vaticaffinol, a resveratrol tetramer isolated from Vatica mangachapoi, on T lymphocytes both in vitro and in vivo, and further explored its potential molecular mechanism. Resveratrol had a wide spectrum of healthy beneficial effects with multiple targets. Interestingly, its tetramer, vaticaffinol, exerted more intensive immunosuppressive activity than resveratrol. Vaticaffinol significantly inhibited T cells proliferation activated by concanavalin A (Con A) or anti-CD3 plus anti-CD28 in a dose- and time-dependent manner. It also induced Con A-activated T cells undergoing apoptosis through mitochondrial pathway. Moreover, this compound prevented cells from entering S phase and G2/M phase during T cells activation. In addition, vaticaffinol inhibited ERK and AKT signaling pathways in Con A-activated T cells. Furthermore, vaticaffinol significantly ameliorated ear swelling in a mouse model of picryl chloride-induced ear contact dermatitis in vivo. In most of the aforementioned experiments, however, resveratrol had only slight effects on the inhibition of T lymphocytes compared with vaticaffinol. Taken together, our findings suggest that vaticaffinol exerts more preferable immunosuppressive activity than its precursor resveratrol both in vitro and in vivo by affecting multiple targets against activated T cells. - Graphical abstract: Vaticaffinol, a resveratrol tetramer isolated from Vatica mangachapoi, exerts more intensive immunosuppressive activity than its precursor resveratrol does in vitro and in vivo. Its mechanism may involve multiple effects against activated T cells: regulation of signalings involved in cell proliferation, G0/G1 arrest of T cells, as well as an apoptosis induction in activated effector T cells. Highlights: ? Vaticaffinol, a resveratrol tetramer, exerts more potent activity than its precursor. ? It inhibited T cells proliferation and prevented them from entering cell cycles. ? It led to apoptosis of activated T cells through mitochondrial pathway. ? It down-regulated ERK and AKT signaling pathways in Con A-activated T cells. ? It significantly ameliorated picryl chloride-induced ear swelling.

  14. Phosphorylation of Spo0A activates its stimulation of in vitro transcription from the Bacillus subtilis spoIIG operon.

    PubMed

    Bird, T H; Grimsley, J K; Hoch, J A; Spiegelman, G B

    1993-08-01

    The spoIIG operon of Bacillus subtilis codes for a sporulation-specific sigma factor, sigma E. In vivo expression of the spoIIG promoter is activated shortly after the onset of sporulation and is dependent on kinA, spo0F, spo0B and spo0A genes. The products of these genes have been shown to participate in a phosphorelay reaction in vitro, culminating in phosphorylation of the transcription factor, Spo0A. The effect of Spo0A phosphorylation on in vitro transcription from the spoIIG promoter was determined. Aliquots from phosphorelay reactions enhanced spoIIG promoter activity 10-fold in transcription assays and stimulation of transcription was dependent on Spo0A phosphorylation. Our results provide biochemical evidence that Spo0A and the phosphorelay form a signal transduction pathway which activates spoII gene expression in development. PMID:8231806

  15. Artefactual nanoparticle activation of the inflammasome platform: in vitro evidence with a nano-formed calcium phosphate

    PubMed Central

    Pele, Laetitia; Haas, Carolin T; Hewitt, Rachel; Faria, Nuno; Brown, Andy; Powell, Jonathan

    2015-01-01

    Aim To determine whether in vitro experimental conditions dictate cellular activation of the inflammasome by apatitic calcium phosphate nanoparticles. Material & methods The responses of blood-derived primary human cells to in situ-formed apatite were investigated under different experimental conditions to assess the effect of aseptic culture, cell rest and duration of particle exposure. Cell death and particle uptake were assessed, while IL-1? and caspase 1 responses, with and without lipopolysaccharide prestimulation, were evaluated as markers of inflammasome activation. Results Under carefully addressed experimental conditions, apatitic nanoparticles did not induce cell death or engage the inflammasome platform, although both could be triggered through artefacts of experimentation. Conclusion In vitro studies often predict that engineered nanoparticles, such as synthetic apatite, are candidates for inflammasome activation and, hence, are toxic. However, the experimental setting must be very carefully considered as it may promote false-positive outcomes. PMID:24991724

  16. Critical analysis of current methods for assessing the in vitro antioxidant and antibacterial activity of plant extracts.

    PubMed

    Tan, Joash Ban Lee; Lim, Yau Yan

    2015-04-01

    Natural product research is an active branch of science, driven by the increased value placed on individual health and well-being. Many naturally-occurring phytochemicals in plants, fruits and vegetables have been reported to exhibit antioxidant and antibacterial activity; often touted as being beneficial for human health. In vitro screening is a common practice in many research laboratories as a means of rapidly assessing these properties. However, the methods used by many are not necessarily optimal; a result of poor standardization, redundant assays and/or outdated methodology. This review primarily aims to give a better understanding in the selection of in vitro assays, with emphasis placed on some common assays such as the total phenolic content assay, free radical scavenging activity, disc-diffusion and broth microdilution. This includes a discussion on the reasons for choosing a particular assay, its strengths and weaknesses, ways to improve the accuracy of results and alternative assays. PMID:25442625

  17. Antidiabetic Activity of Ruellia tuberosa L., Role of ?-Amylase Inhibitor: In Silico, In Vitro, and In Vivo Approaches

    PubMed Central

    Ratna Wulan, Dyah; Priyo Utomo, Edi; Mahdi, Chanif

    2015-01-01

    Ruellia tuberosa L. is a folk remedy in the treatment of diabetes mellitus. However, its hypoglycemic activity has not been investigated so far. In the present study, the antidiabetic mechanism of the n-hexane fraction of methanolic extract (HFME) of this plant was investigated in silico, in vitro, and in vivo. In silico study was performed using AutoDock4.2 software. In vitro???-amylase inhibitory activity was investigated by starch-iodine method. A single dose of 450?mg/kg HFME for 14 days was subjected to an antidiabetic screening in vivo by a multiple low dose streptozotocin (MLD-STZ) induced rats. Molecular modeling results show that Betulin exhibited noncompetitive ?-amylase inhibitory activities. The effect of HFME elicited significant reductions of diabetic rat blood glucose. A single dose administration of HFME inhibited ?-amylase activity in vivo (P < 0.01) compared to a diabetic control group. Moreover, this extract strongly inhibited the ?-amylase activity in vitro (IC50 0.14 ± 0.005?mg/mL). It is concluded that HFME exerted an antidiabetic effect via ?-amylase inhibitor. Our findings provide a possible hypoglycemic action of R. tuberosa L. as an alternative therapy in the management of diabetes. PMID:26576302

  18. In Vitro Inhibition Of Three Different Drugs Used In Rheumatoid Arthritis Treatment On Human Serum Paraoxanase 1 Enzyme Activity.

    PubMed

    Dilek, Esra; Polat, M Fatih

    2016-01-01

    We studied in vitro effects of three different drugs (ibuprofen, meloxicam and methotrexate) which are often used in rheumatoid arthritis (RA) treatment on human serum paraoxanase1 (PON1) enzyme activity. The drugs used in RA treatment decreased the in vitro PON1 activity. The inhibition mechanism of ibuprofen and methotrexate were noncompetitive whereas meloxicam was a competitive inhibitor. The IC50 values for ibuprofen, meloxicam and methotrexate were calculated to be 0.35 mM, 0.10 mM, and 0.18 mM, respectively, and the Ki constants were calculated to be 0.890 mM, 0.125 mM, and 0.260 mM, respectively. The IC50 and Ki values showed the maximum inhibition of meloxicam drugs. We propose a prediction scheme for the interaction of meloxicam with the PON1 active site because we thought that meloxicam interacts with the amino acids which are in the PON1 enzyme active site. The results we found showed that these drugs which are often used in RA treatment in vitro inhibit the activity of the enzyme with different inhibition mechanisms at low doses. PMID:26458405

  19. Carbon-activated gas filtration during in vitro culture increased pregnancy rate following transfer of in vitro-produced bovine embryos.

    PubMed

    Merton, J S; Vermeulen, Z L; Otter, T; Mullaart, E; de Ruigh, L; Hasler, J F

    2007-04-15

    Many environmental conditions for in vitro embryo production (IVP) systems for cattle have been relatively standardised, e.g. media composition, temperature, pH, water quality, and atmospheric composition. However, little attention has been paid to the quality of ambient laboratory air and the gas environment in incubators. Although a few studies have examined the effects of chemical air contamination on IVP of human embryos, there are no published accounts for domestic animal embryos. Therefore, this study investigated the effects of an intra-incubator carbon-activated air filtration system (CODA) during in vitro culture (IVC) on embryonic development and subsequent pregnancy rate of bovine embryos. Immature cumulus-oocyte-complexes (COCs) were obtained twice-weekly by ultrasonic-guided transvaginal oocyte aspiration. The COCs were matured in TCM199/FCS/LH/FSH, fertilized with frozen-thawed Percoll-separated semen, and subsequently cultured for 7 day in SOFaaBSA. Day 7 embryos were transferred either fresh or frozen/thawed. The experimental design was a 2 x 2 factorial; presumptive zygotes were placed either in a conventional CO(2)-O(2)-N(2) incubator (Control group) or in an identical CO(2)-O(2)-N(2) incubator with a CODA intra-incubator air purification unit (CODA group) for IVC. The embryo production rate at Day 7 was not affected by the CODA air purification unit (23.4 and 24.7% morulae and blastocysts per oocyte for control and CODA, respectively) nor was there any significant effect on embryo stage or quality. However, the pregnancy rate was improved (P=0.043) for both fresh (46.3% versus 41.0%) and frozen/thawed embryos (40.8% versus 35.6%). In conclusion, atmospheric purification by the CODA intra-incubator air purification unit significantly increased pregnancy rate following transfer of in vitro-produced bovine embryos. PMID:17331571

  20. Pentoxifylline inhibits Vgamma9/Vdelta2 T lymphocyte activation of patients with active Behçets disease in vitro.

    PubMed

    Accardo-Palumbo, A; Ferrante, A; Ciccia, F; Cadelo, M; Giardina, A R; Impastato, R; Triolo, G

    2007-01-01

    The aim of this study is to evaluate the in vitro effect of pentoxifylline (PTX) on T Vgamma9/Vdelta2 lymphocyte function in Behçets disease (BD). We investigated the effect of PTX on Vgamma9/Vdelta2 T cell expansion and expression of TNFRII receptor and perforin content before and after PTX addition by means of FACS analysis lymphocyte cultures from patients with active and inactive BD and healthy subjects. The addition of PTX at a concentration of 1 mg/ml determined a significant inhibition of cell expansion, a down regulation of TNF receptor expression and inhibited the PMA-induced degranulation of perforin. Taken together these data indicate that PTX is capable of interfering with Vgamma9/Vdelta2 T cell function in BD, and although cell culture models cannot reliably predict all of the potential effects of the drug in vivo, our results encourage the possibility that this drug may find use in a range of immunological disorder characterized by dysregulated cell-mediated immunity. PMID:17880773

  1. Applicator for in-vitro ultrasound-activated targeted drug delivery

    NASA Astrophysics Data System (ADS)

    Gerold, B.; Gourevich, D.; Volovick, A.; Xu, D.; Arditti, F.; Prentice, P.; Cochran, S.; Gnaim, J.; Medan, Y.; Wang, L.; Melzer, A.

    2012-10-01

    Reducing toxicity and improving uptake of cancer drugs in tumors are important goals of targeted drug delivery (TDD). Ultrasonic drug release from various encapsulants has been a focus of many research groups. However, a single standard ultrasonic device, viable for use by biologists, is not currently present in the market. The device reported here is designed to allow investigation of the impact of ultrasound on cellular uptake and cell viability in-vitro. In it, single-element transducers with different operating frequencies are mounted below a standard 96-well plate. The plate is moved above the transducers, such that each line of wells can be sonicated at a different frequency. To assess the device, 96-well plates were seeded with cells and sonicated using different ultrasonic parameters, with and without doxorubicin. Cell viability was measured by colorimetric MTT assay and the uptake of doxorubicin by cells was also determined. The device proved to be highly viable in preliminary tests; it demonstrated that change in ultrasonic parameters produces different effect on cells. For example, increase in uptake of doxorubicin was demonstrated following ultrasound application. The growing interest in ultrasound-activated TDD emphasizes the need for standardization of the ultrasound device and the one reported here may offer some indications of how that may be achieved. It is planned to further improve the prototype by increasing the number of ultrasonic frequencies and degrees of freedom for each transducer.

  2. A Class of 5-Nitro-2-furancarboxylamides with Potent Trypanocidal Activity against Trypanosoma brucei in Vitro

    PubMed Central

    2013-01-01

    Recently, the World Health Organization approved the nifurtimox–eflornithine combination therapy for the treatment of human African trypanosomiasis, renewing interest in nitroheterocycle therapies for this and associated diseases. In this study, we have synthesized a series of novel 5-nitro-2-furancarboxylamides that show potent trypanocidal activity, ?1000-fold more potent than nifurtimox against in vitro Trypanosoma brucei with very low cytotoxicity against human HeLa cells. More importantly, the most potent analogue showed very limited cross-resistance to nifurtimox-resistant cells and vice versa. This implies that our novel, relatively easy to synthesize and therefore cheap, 5-nitro-2-furancarboxylamides are targeting a different, but still essential, biochemical process to those targeted by nifurtimox or its metabolites in the parasites. The significant increase in potency (smaller dose probably required) has the potential for greatly reducing unwanted side effects and also reducing the likelihood of drug resistance. Collectively, these findings have important implications for the future therapeutic treatment of African sleeping sickness. PMID:23281892

  3. Activity of Porphyridium sp. polysaccharide against herpes simplex viruses in vitro and in vivo.

    PubMed

    Huheihel, Mahmoud; Ishanu, Vladimir; Tal, Jacov; Arad, Shoshana Malis

    2002-01-01

    The cell wall sulfated polysaccharide of the red microalga Porphyridium sp. exhibited impressive antiviral activity against herpes simplex virus types 1 and 2 (HSV-1 and -2) both in vitro (cell culture) and in vivo (rats and rabbits). Depending on the concentration, this polysaccharide completely inhibited or slowed down the development of the cytopathic effect in HSV-infected cells, but did not show any cytotoxic effects on vero cells even when a concentration as high as 250 microg/ml was used. There was indirect evidence for a strong interaction between the polysaccharide and HSV and a weak interaction with the cell surface. When tested in vivo, Porphyridium sp. polysaccharide conferred significant and efficient protection against HSV-1 infection: at a concentration as low as 100 microg/ml, it prevented the appearance and development of symptoms of HSV-1 infection in rats and rabbits. The polysaccharide did not exhibit any cytotoxic effects at a concentration of 2 mg/ml in vivo. PMID:11741707

  4. Ultrasonic-assisted extraction and in-vitro antioxidant activity of polysaccharide from Hibiscus leaf.

    PubMed

    Afshari, Kasra; Samavati, Vahid; Shahidi, Seyed-Ahmad

    2015-03-01

    The effects of ultrasonic power, extraction time, extraction temperature, and the water-to-raw material ratio on extraction yield of crude polysaccharide from the leaf of Hibiscus rosa-sinensis (HRLP) were optimized by statistical analysis using response surface methodology. The response surface methodology (RSM) was used to optimize HRLP extraction yield by implementing the Box-Behnken design (BBD). The experimental data obtained were fitted to a second-order polynomial equation using multiple regression analysis and also analyzed by appropriate statistical methods (ANOVA). Analysis of the results showed that the linear and quadratic terms of these four variables had significant effects. The optimal conditions for the highest extraction yield of HRLP were: ultrasonic power, 93.59 W; extraction time, 25.71 min; extraction temperature, 93.18°C; and the water to raw material ratio, 24.3 mL/g. Under these conditions, the experimental yield was 9.66±0.18%, which is well in close agreement with the value predicted by the model 9.526%. The results demonstrated that HRLP had strong scavenging activities in vitro on DPPH and hydroxyl radicals. PMID:25150112

  5. In vitro assessment of plant lectins with anti-pinwood nematode activity.

    PubMed

    Gaofu, Qi; Shiqing, Mao; Fayin, Zhu; Zhiniu, Yu; Xiuyun, Zhao

    2008-05-01

    Two lectin proteins were purified from the corms of Pinellia ternata and Lycoris radiata. Both P. ternata agglutinin (PTA) protein and L. radiata agglutinin (LRA) protein formed polymers and coagulated both rabbit red blood cells and yeast cells. The two proteins were each diluted to different concentration and then mixed with pinewood nematodes, and nematode survival was measured. Results showed that the two lectin proteins showed significant levels of resistance against nematodes and the nematode population was significantly reduced, compared to PBS buffer without protein control group. The mean number of nematodes of two lectin proteins group was significantly lower than that of control group constantly throughout the assay period with differences being very significant at P<0.01 after 24 h. After 96 h, when 500 microg/ml proteins were used, nematode number significantly declined to an average of 26 (approximately 43% of the controls) and 32.2 (approximately 53.3% of the controls) nematodes at LRA and PTA protein, respectively, compared to the control group. Results also indicated that higher concentrations of protein were more toxic to the pinewood nematode. Even when the concentration was as low as 30 microg/ml, the toxic proteins retained their anti-nematode activity. Furthermore, pinewood nematode was exposed to the proteins for longer, more pinewood nematodes were killed. Our results indicated the two lectin proteins both apparently have a toxic effect on the pinewood nematode that affects its survival in vitro. PMID:18158158

  6. Fabrication and In Vitro Deployment of a Laser-Activated Shape Memory Polymer Vascular Stent

    SciTech Connect

    Baer, G M; Small IV, W; Wilson, T S; Benett, W J; Matthews, D L; Hartman, J; Maitland, D J

    2007-04-25

    Vascular stents are small tubular scaffolds used in the treatment of arterial stenosis (narrowing of the vessel). Most vascular stents are metallic and are deployed either by balloon expansion or by self-expansion. A shape memory polymer (SMP) stent may enhance flexibility, compliance, and drug elution compared to its current metallic counterparts. The purpose of this study was to describe the fabrication of a laser-activated SMP stent and demonstrate photothermal expansion of the stent in an in vitro artery model. A novel SMP stent was fabricated from thermoplastic polyurethane. A solid SMP tube formed by dip coating a stainless steel pin was laser-etched to create the mesh pattern of the finished stent. The stent was crimped over a fiber-optic cylindrical light diffuser coupled to an infrared diode laser. Photothermal actuation of the stent was performed in a water-filled mock artery. At a physiological flow rate, the stent did not fully expand at the maximum laser power (8.6 W) due to convective cooling. However, under zero flow, simulating the technique of endovascular flow occlusion, complete laser actuation was achieved in the mock artery at a laser power of {approx}8 W. We have shown the design and fabrication of an SMP stent and a means of light delivery for photothermal actuation. Though further studies are required to optimize the device and assess thermal tissue damage, photothermal actuation of the SMP stent was demonstrated.

  7. In vitro evaluation of the antimicrobial activity of chlorhexidine and sodium hypochlorite.

    PubMed

    Vianna, Morgana Eli; Gomes, Brenda P F A; Berber, Vanessa Bellocchio; Zaia, Alexandre Augusto; Ferraz, Caio Cezar Randi; de Souza-Filho, Francisco José

    2004-01-01

    The aim of this study was to investigate in vitro the antimicrobial activity of 0.2%, 1%, and 2% chlorhexidine gluconate (CHX gel and CHX liquid), against endodontic pathogens and compare the results with the ones achieved by 0.5%, 1%, 2.5%, 4%, and 5.25% sodium hypochlorite (NaOCl). A broth dilution test was performed, and the timing for irrigants to kill microbial cells was recorded and statistically analyzed. Both 2.0% gel and liquid formulations eliminated Staphylococcus aureus and Candida albicans in 15 seconds, whereas the gel formulation killed Enterococcus faecalis in 1 minute. All tested irrigants eliminated Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella intermedia in 15 seconds. The timing required for 1.0% and 2.0% CHX liquid to eliminate all microorganisms was the same required for 5.25% NaOCl. The antimicrobial action is related to type, concentration, and presentation form of the irrigants as well as the microbial susceptibility. PMID:14716261

  8. In Vitro and In Vivo Antitumor Activity of a Novel Semisynthetic Derivative of Cucurbitacin B

    PubMed Central

    Silva, Izabella T.; Carvalho, Annelise; Lang, Karen L.; Dudek, Sabine E.; Masemann, Dörthe; Durán, Fernando J.; Caro, Miguel S. B.; Rapp, Ulf R.; Wixler, Viktor; Schenkel, Eloir P.; Simőes, Cláudia M. O.; Ludwig, Stephan

    2015-01-01

    Lung cancer is the most deadly type of cancer in humans, with non-small-cell lung cancer (NSCLC) being the most frequent and aggressive type of lung cancer showing high resistance to radiation and chemotherapy. Despite the outstanding progress made in anti-tumor therapy, discovering effective anti-tumor drugs is still a challenging task. Here we describe a new semisynthetic derivative of cucurbitacin B (DACE) as a potent inhibitor of NSCLC cell proliferation. DACE arrested the cell cycle of lung epithelial cells at the G2/M phase and induced cell apoptosis by interfering with EGFR activation and its downstream signaling, including AKT, ERK, and STAT3. Consistent with our in vitro studies, intraperitoneal application of DACE significantly suppressed the growth of mouse NSCLC that arises from type II alveolar pneumocytes due to constitutive expression of a human oncogenic c-RAF kinase (c-RAF-1-BxB) transgene in these cells. Taken together, these findings suggest that DACE is a promising lead compound for the development of an anti-lung-cancer drug. PMID:25674792

  9. In Vitro and In Vivo Trypanocidal Activity of H2bdtc-Loaded Solid Lipid Nanoparticles

    PubMed Central

    Carneiro, Zumira A.; da S. Maia, Pedro I.; Sesti-Costa, Renata; Lopes, Carla D.; Pereira, Tatiana A.; Milanezi, Cristiane M.; da Silva, Marcelo A. Pereira.; Lopez, Renata F. V.; Silva, Joăo S.; Deflon, Victor M.

    2014-01-01

    The parasite Trypanosoma cruzi causes Chagas disease, which remains a serious public health concern and continues to victimize thousands of people, primarily in the poorest regions of Latin America. In the search for new therapeutic drugs against T. cruzi, here we have evaluated both the in vitro and the in vivo activity of 5-hydroxy-3-methyl-5-phenyl-pyrazoline-1-(S-benzyl dithiocarbazate) (H2bdtc) as a free compound or encapsulated into solid lipid nanoparticles (SLN); we compared the results with those achieved by using the currently employed drug, benznidazole. H2bdtc encapsulated into solid lipid nanoparticles (a) effectively reduced parasitemia in mice at concentrations 100 times lower than that normally employed for benznidazole (clinically applied at a concentration of 400 µmol kg?1 day?1); (b) diminished inflammation and lesions of the liver and heart; and (c) resulted in 100% survival of mice infected with T. cruzi. Therefore, H2bdtc is a potent trypanocidal agent. PMID:24810753

  10. In vitro and in vivo trypanocidal activity of H2bdtc-loaded solid lipid nanoparticles.

    PubMed

    Carneiro, Zumira A; Maia, Pedro I da S; Sesti-Costa, Renata; Lopes, Carla D; Pereira, Tatiana A; Milanezi, Cristiane M; da Silva, Marcelo A Pereira; Lopez, Renata F V; Silva, Joăo S; Deflon, Victor M

    2014-05-01

    The parasite Trypanosoma cruzi causes Chagas disease, which remains a serious public health concern and continues to victimize thousands of people, primarily in the poorest regions of Latin America. In the search for new therapeutic drugs against T. cruzi, here we have evaluated both the in vitro and the in vivo activity of 5-hydroxy-3-methyl-5-phenyl-pyrazoline-1-(S-benzyl dithiocarbazate) (H2bdtc) as a free compound or encapsulated into solid lipid nanoparticles (SLN); we compared the results with those achieved by using the currently employed drug, benznidazole. H2bdtc encapsulated into solid lipid nanoparticles (a) effectively reduced parasitemia in mice at concentrations 100 times lower than that normally employed for benznidazole (clinically applied at a concentration of 400 µmol kg(-1) day(-1)); (b) diminished inflammation and lesions of the liver and heart; and (c) resulted in 100% survival of mice infected with T. cruzi. Therefore, H2bdtc is a potent trypanocidal agent. PMID:24810753

  11. DMSO containing ruthenium(ii) hydrazone complexes: in vitro evaluation of biomolecular interaction and anticancer activity.

    PubMed

    Alagesan, M; Sathyadevi, P; Krishnamoorthy, P; Bhuvanesh, N S P; Dharmaraj, N

    2014-11-14

    Synthesis, spectral, electrochemical and single crystal X-ray diffraction data of a new series of DMSO containing bivalent ruthenium hydrazone complexes are presented. XRD data of two of the new complexes revealed an octahedral coordination around the ruthenium ion satisfied by NOS2Cl2 atoms. Electrochemical studies showed the metal centred, quasi-reversible, one-electron redox behaviour of the new complexes. The binding of these complexes with biomolecules such as calf thymus DNA (CT-DNA) and bovine serum albumin (BSA) protein investigated by different spectrophotometric methods revealed an intercalative mode of interaction. The in vitro cytotoxicity of these complexes evaluated by the MTT assay on a panel of cancer and normal cell lines indicated that the above complexes are more toxic to cancer cells with a few micromolar concentrations as the IC50 value, but are significantly less toxic to normal cell lines. The observed variations in the binding interactions and cytotoxicity of the complexes were attributed to the nature of the hydrazide moiety of the hydrazones that influences their biological activities. PMID:25223849

  12. Luteolin nanoparticle in chemoprevention – in vitro and in vivo anticancer activity

    PubMed Central

    Majumdar, Debatosh; Jung, Kyung-Ho; Zhang, Hongzheng; Nannapaneni, Sreenivas; Wang, Xu; Amin, A.R.M Ruhul; Chen, Zhengjia; Chen, Zhuo (G).; Shin, Dong M.

    2013-01-01

    Cancer prevention (chemoprevention) by using naturally occurring dietary agents has gained immense interest due to the broad safety window of these compounds. However, many of these compounds are hydrophobic and poorly soluble in water. They frequently display low bioavailability, poor systemic delivery, and low efficacy. To circumvent this problem, we explored a novel approach towards chemoprevention using nanotechnology to deliver luteolin, a natural compound present in green vegetables. We formulated water soluble polymer-encapsulated Nano-Luteolin from hydrophobic luteolin, and studied its anticancer activity against lung cancer and head and neck cancer. In vitro studies demonstrated that, like luteolin, Nano-Luteolin inhibited the growth of lung cancer cells (H292 cell line) and squamous cell carcinoma of head and neck (SCCHN) cells (Tu212 cell line). In Tu212 cells, the IC50 value of Nano-Luteolin was 4.13?M, and that of luteolin was 6.96?M. In H292 cells, the IC50 of luteolin was 15.56?M, and Nano-Luteolin was 14.96?M. In vivo studies using a tumor xenograft mouse model demonstrated that Nano-Luteolin has a significant inhibitory effect on the tumor growth of SCCHN in comparison to luteolin. Our results suggest that nanoparticle delivery of naturally occurring dietary agents like luteolin has many advantages and may have potential application in chemoprevention in clinical settings. PMID:24403290

  13. Comparative in vitro study of the antimicrobial activities of different commercial antibiotic products for intravenous administration

    PubMed Central

    2010-01-01

    Background The antimicrobial resistance is a global problem, probably due to the indiscriminate and irrational use of antibiotics, prescriptions for incorrect medicines or incorrect determinations of dose, route and/or duration. Another consideration is the uncertainty of patients receiving antibiotics about whether the quality of a generic medicine is equal to, greater than or less than its equivalent brand-name drug. The antibiotics behaviors must be evaluated in vitro and in vivo in order to confirm their suitability for therapeutic use. Methods The antimicrobial activities of Meropenem and Piperacillin/Tazobactam were studied by microbiological assays to determine their potencies (content), minimal inhibitory concentrations (MICs), critical concentrations and capacity to produce spontaneous drug-resistant mutants. Results With respect to potency (content) all the products fulfill USP requirements, so they should all be considered pharmaceutical equivalents. The MIC values of the samples evaluated (trade marks and generics) were the same for each strain tested, indicating that all products behaved similarly. The critical concentration values were very similar for all samples, and the ratios between the critical concentration of the standard and those of each sample were similar to the ratios of their specific antibiotic contents. Overall, therefore, the results showed no significant differences among samples. Finally, the production of spontaneous mutants did not differ significantly among the samples evaluated. Conclusions All the samples are pharmaceutical equivalents and the products can be used in antimicrobial therapy. PMID:20113478

  14. In vitro assessment of the antimicrobial activity of new N-acyl-thiourea derivatives.

    PubMed

    Di?u, Lia Mara; Mih?escu, Gr; Chifiriuc, Carmen; Bleotu, Coralia; Morusciag, Lauren?iu; Ni?ulescu, George Mihai; Missir, Alexandru

    2010-01-01

    The qualitative screening of the susceptibility spectra of different microbial strains to the newly synthesized substances complexes was performed by adapted disk diffusion techniques, while the quantitative assay of the minimal inhibitory concentration (M.I.C., microg/cm3) value was based on liquid medium serial microdilutions. The compounds were solubilized in dimethylsulfoxide (DMSO). The in vitro biological screening effects were tested against a microbial inoculum of approximately 1.5 x 10(8) UFC/cm3, corresponding to 0.5 McFarland standard density, obtained from Gram positive (Staphylococcus aureus, Bacillus subtilis), Gram negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa) and fungal strains (Candida albicans). In order to investigate the influence of the subinhibitory concentration of the tested substances on the expression of different virulence features, the strains were incubated overnight in the presence of the newly synthesized thiourea derivatives (vol:vol) and different virulence features were investigated, i.e: adherence capacity to the cellular substrate represented by HeLa cells and to inert substrata quantified by slime test and soluble enzymatic virulence factors (haemolysins and other pore-forming toxins, proteases activity, DN-ase and siderophores production). The cytotoxicity was assessed microscopically, by observing the effect of the tested compounds on the cellular substratum integrity. PMID:21053783

  15. In vitro anticancer activities of Schiff base and its lanthanum complex.

    PubMed

    Neelima; Poonia, Kavita; Siddiqui, Sahabjada; Arshad, Md; Kumar, Dinesh

    2016-02-15

    Schiff base metal complexes are well-known to intercalate DNA. The La(III) complexes have been synthesized such that they hinder with the role of the topoisomerases, which control the topology of DNA during the cell-division cycle. Although several promising chemotherapeutics have been developed, on the basis of Schiff base metal complex DNA intercalating system they did not proceed past clinical trials due to their dose-limiting toxicity. Herein, we discuss an alternative compound, the La(III) complex, [La(L(1))2Cl3]·7H2O based on a Schiff base ligand 2,3-dihydro-1H-indolo-[2,3-b]-phenazin-4(5H)-ylidene)benzothiazole-2-amine (L(1)), and report in vitro cell studies. Results of antitumor activity using cell viability assay, reactive oxygen species (ROS) generation and nuclear condensation in PC-3 (Human, prostate carcinoma) cells show that the metal complex is more potent than ligand. La(III) complexes have been synthesized by reaction of lanthanum(III) salt in 1:2M ratio with ligands L(1) and 3-(ethoxymethylene)-2,3-dihydro-1H-indolo[2,3-b]-phenazin-4(5H)-ylidene)benzathiazole-2-amine (L(2)) in methanol. The ligands and their La(III) complexes were characterized by molar conductance, magnetic susceptibility, elemental analyses, FT-IR, UV-Vis, (1)H/(13)C NMR, thermogravimetric, XRD, and SEM analysis. PMID:26619196

  16. In vitro characterization and mosquito (Aedes aegypti) repellent activity of essential-oils-loaded nanoemulsions.

    PubMed

    Nuchuchua, Onanong; Sakulku, Usawadee; Uawongyart, Napaporn; Puttipipatkhachorn, Satit; Soottitantawat, Apinan; Ruktanonchai, Uracha

    2009-01-01

    The nanoemulsions composed of citronella oil, hairy basil oil, and vetiver oil with mean droplet sizes ranging from 150 to 220 nm were prepared and investigated both in vitro and in vivo. Larger emulsion droplets (195-220 nm) shifted toward a smaller size (150-160 nm) after high-pressure homogenization and resulted in higher release rate. We proposed that thin films obtained from the nanoemulsions with smaller droplet size would have higher integrity, thus increasing the vaporization of essential oils and subsequently prolonging the mosquito repellant activity. The release rates were fitted with Avrami's equations and n values were in the same range of 0.6 to 1.0, implying that the release of encapsulated limonene was controlled by the diffusion mechanism from the emulsion droplet. By using high-pressure homogenization together with optimum concentrations of 5% (w/w) hairy basil oil, 5% (w/w) vetiver oil (5%), and 10% (w/w) citronella oil could improve physical stability and prolong mosquito protection time to 4.7 h due to the combination of these three essential oils as well as small droplet size of nanoemulsion. PMID:19862624

  17. In vivo and in vitro studies on the anticancer activity of Copaifera multijuga hayne and its fractions.

    PubMed

    Lima, Sylvia R M; Junior, Valdir F Veiga; Christo, Herick B; Pinto, Angelo C; Fernandes, Patricia D

    2003-11-01

    Copaiba oil resin (COR) obtained from Copaifera multijuga Hayne has been used in popular medicine as an antinflammatory and for the treatment of bronchitis, ulcers and cancer. The aim of this study was to evaluate the action of COR and its fractions on the inhibition of lung metastasis and tumour growth induced by B16F10 melanoma cells in mice and cytotoxicity in vitro using Trypan Blue exclusion method and MTT conversion. Mice which have received subcutaneously B16F10 cells developed a solid tumour that reached a peak at 17 days. Together with the increase in tumour growth we also observed an increase in the number of lung nodules. There was a positive correlation between the in vitro cytotoxic assay and in vivo antitumour activity. The oral administration of COR (at 2 g/Kg in the days 3, 5, 7, 10, 12 and 14 after inoculation of tumoral cells) reduced tumour growth by 58% and tumour weight by 76%. At the same dose COR reduced the number of lung nodules by 47.1%. In vitro experiments showed that COR incubated with the melanoma cell line reduced cell viability in a concentration and time-dependent manner. Diterpenic and sesquiterpenic fractions or reconstituted oil induced cytotoxicity. Our results shows that COR and its fractions have tumouricidal activity in the melanoma cell line in both models in vivo and in vitro. PMID:14595585

  18. Ttyh1 protein is expressed in glia in vitro and shows elevated expression in activated astrocytes following status epilepticus.

    PubMed

    Wiernasz, Elzbieta; Kaliszewska, Aleksandra; Brutkowski, Wojciech; Bednarczyk, Joanna; Gorniak, Malgorzata; Kaza, Beata; Lukasiuk, Katarzyna

    2014-12-01

    In a previous study, we showed that Ttyh1 protein is expressed in neurons in vitro and in vivo in the form of punctuate structures, which are localized to neuropil and neuronal somata. Herein, we provide the first description of Ttyh1 protein expression in astrocytes, oligodendrocytes and microglia in vitro. Moreover, using double immunofluorescence, we show Ttyh1 protein expression in activated astrocytes in the hippocampus following amygdala stimulation-induced status epilepticus. We demonstrate that in migrating astrocytes in in vitro wound model Ttyh1 concentrates at the edges of extending processes. These data suggest that Ttyh1 not only participates in shaping neuronal morphology, as previously described, but may also play a role in the function of activated glia in brain pathology. To localize Ttyh1 expression in the cellular compartments of neurons and astrocytes, we performed in vitro double immunofluorescent staining using markers for the following subcellular structures: endoplasmic reticulum (GRP78), Golgi apparatus (GM130), clathrin-coated vehicles (clathrin), early endosomes (Rab5 and APPL2), recycling endosomes (Rab11), trans-Golgi network (TGN46), endoplasmic reticulum membrane (calnexin), late endosomes and lysosomes (LAMP1) and synaptic vesicles (synaptoporin and synaptotagmin 1). We found that Ttyh1 is present in the endoplasmic reticulum, Golgi apparatus and clathrin-coated vesicles (clathrin) in both neurons and astrocytes and also in late endosomes or lysosomes in astrocytes. The presence of Ttyh1 was negligible in early endosomes, recycling endosomes, trans-Golgi network, endoplasmic reticulum membrane and synaptic vesicles. PMID:25316497

  19. A highly sensitive fluorescent indicator dye for calcium imaging of neural activity in vitro and in vivo

    PubMed Central

    Tada, Mayumi; Takeuchi, Atsuya; Hashizume, Miki; Kitamura, Kazuo; Kano, Masanobu

    2014-01-01

    Calcium imaging of individual neurons is widely used for monitoring their activity in vitro and in vivo. Synthetic fluorescent calcium indicator dyes are commonly used, but the resulting calcium signals sometimes suffer from a low signal-to-noise ratio (SNR). Therefore, it is difficult to detect signals caused by single action potentials (APs) particularly from neurons in vivo. Here we showed that a recently developed calcium indicator dye, Cal-520, is sufficiently sensitive to reliably detect single APs both in vitro and in vivo. In neocortical neurons, calcium signals were linearly correlated with the number of APs, and the SNR was > 6 for in vitro slice preparations and > 1.6 for in vivo anesthetised mice. In cerebellar Purkinje cells, dendritic calcium transients evoked by climbing fiber inputs were clearly observed in anesthetised mice with a high SNR and fast decay time. These characteristics of Cal-520 are a great advantage over those of Oregon Green BAPTA-1, the most commonly used calcium indicator dye, for monitoring the activity of individual neurons both in vitro and in vivo. PMID:24405482

  20. 1950 MHz IMT-2000 field does not activate microglial cells in vitro.

    PubMed

    Hirose, Hideki; Sasaki, Atsushi; Ishii, Nana; Sekijima, Masaru; Iyama, Takahiro; Nojima, Toshio; Ugawa, Yoshikazu

    2010-02-01

    Given the widespread use of the cellular phone today, investigation of potential biological effects of radiofrequency (RF) fields has become increasingly important. In particular, much research has been conducted on RF effects on brain function. To examine any biological effects on the central nervous system (CNS) induced by 1950 MHz modulation signals, which are controlled by the International Mobile Telecommunication-2000 (IMT-2000) cellular system, we investigated the effect of RF fields on microglial cells in the brain. We assessed functional changes in microglial cells by examining changes in immune reaction-related molecule expression and cytokine production after exposure to a 1950 MHz Wideband Code Division Multiple Access (W-CDMA) RF field, at specific absorption rates (SARs) of 0.2, 0.8, and 2.0 W/kg. Primary microglial cell cultures prepared from neonatal rats were subjected to an RF or sham field for 2 h. Assay samples obtained 24 and 72 h after exposure were processed in a blind manner. Results showed that the percentage of cells positive for major histocompatibility complex (MHC) class II, which is the most common marker for activated microglial cells, was similar between cells exposed to W-CDMA radiation and sham-exposed controls. No statistically significant differences were observed between any of the RF field exposure groups and the sham-exposed controls in percentage of MHC class II positive cells. Further, no remarkable differences in the production of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6) were observed between the test groups exposed to W-CDMA signal and the sham-exposed negative controls. These findings suggest that exposure to RF fields up to 2 W/kg does not activate microglial cells in vitro. PMID:19650078

  1. Antibacterial, antifungal and in vitro antileukaemia activity of metal complexes with thiosemicarbazones.

    PubMed

    Pahontu, Elena; Julea, Felicia; Rosu, Tudor; Purcarea, Victor; Chumakov, Yurie; Petrenco, Petru; Gulea, Aurelian

    2015-04-01

    1-phenyl-3-methyl-4-benzoyl-5-pyrazolone 4-ethyl-thiosemicarbazone (HL) and its copper(II), vanadium(V) and nickel(II) complexes: [Cu(L)(Cl)]·C?H?OH·(1), [Cu(L)?]·H?O (2), [Cu(L)(Br)]·H?O·CH?OH (3), [Cu(L)(NO?)]·2C?H?OH (4), [VO?(L)]·2H?O (5), [Ni(L)?]·H?O (6), were synthesized and characterized. The ligand has been characterized by elemental analyses, IR, (1) H NMR and (13) C NMR spectroscopy. The tridentate nature of the ligand is evident from the IR spectra. The copper(II), vanadium(V) and nickel(II) complexes have been characterized by different physico-chemical techniques such as molar conductivity, magnetic susceptibility measurements and electronic, infrared and electron paramagnetic resonance spectral studies. The structures of the ligand and its copper(II) (2, 4), and vanadium(V) (5) complexes have been determined by single-crystal X-ray diffraction. The composition of the coordination polyhedron of the central atom in 2, 4 and 5 is different. The tetrahedral coordination geometry of Cu was found in complex 2 while in complex 4, it is square planar, in complex 5 the coordination polyhedron of the central ion is distorted square pyramid. The in vitro antibacterial activity of the complexes against Escherichia coli, Salmonella abony, Staphylococcus aureus, Bacillus cereus and the antifungal activity against Candida albicans strains was higher for the metal complexes than for free ligand. The effect of the free ligand and its metal complexes on the proliferation of HL-60 cells was tested. PMID:25708540

  2. SPME determination of volatile aldehydes for evaluation of in-vitro antioxidant activity.

    PubMed

    Stashenko, Elena E; Puertas, Miguel A; Martínez, Jairo R

    2002-05-01

    The in-vitro antioxidant activity of natural (essential oils, vitamin E) or synthetic substances ( tert-butyl hydroxy anisole (BHA), Trolox) has been evaluated by monitoring volatile carbonyl compounds released in model lipid systems subjected to peroxidation. The procedure employed methodology previously developed for the determination of carbonyl compounds as their pentafluorophenylhydrazine derivatives which were quantified, with high sensitivity, by means of capillary gas chromatography with electron-capture detection. Linoleic acid and sunflower oil were used as model lipid systems. Lipid peroxidation was induced in linoleic acid by the Fe2+ ion (1 mmol L-1, 37 degrees C, 12 h) and in sunflower oil by heating in the presence of O2 (220 degrees C, 2 h). The change in hexanal (the main lipoxidation product) concentration found in the lipid matrix subjected to oxidation with and without the substance being tested was used to calculate the antioxidant protection effect. These procedures were employed to evaluate the antioxidant activity of the essential oils of cilantro ( Coriander sativum L.), fennel ( Foeniculum vulgare Mill.), rosemary ( Rosmarinus officinalis L.), "salvia negra" ( Lepechinia schiedeana), and oregano ( Origanum vulgare L.), and the well-known antioxidants BHA, vitamin E, and Trolox, its water-soluble analog. In the sunflower oil system, the essential oils had a stronger protective effect against lipid peroxidation than BHA, vitamin E, and Trolox within the range of concentrations examined (1-20 g L-1). The highest protecting effect, corresponding to a 90% drop in hexanal release, was observed for cilantro oil at 10 g L-1. PMID:12012174

  3. Antifungal activity of organotin compounds with functionalized carboxylates evaluated by the microdilution bioassay in vitro.

    PubMed

    Dylag, Mariusz; Pruchnik, Hanna; Pruchnik, Florian; Majkowska-Skrobek, Grazyna; U?aszewski, Stanis?aw

    2010-03-01

    We investigated the susceptibility of 96 well-characterized strains of yeast-like and filamentous fungi towards new organotin compounds: (1) [Sn(C4H9)3(OOCC6H4SO3H-2)], (2) Sn(C4H9)3{OOC(CH2)3P(C6H5)3}]Br, and (3) [Sn(C6H5)3[OOC(CH2)3N(CH3)3}]Cl. In the case of yeast-like fungi, the in vitro susceptibility tests were carried out according to the Clinical Laboratory Standards Institute (CLSI, formerly NCCLS) reference method M27-A2, while for filamentous fungi the investigations were conducted according to the M38-A and M38-P methods. The organotin complexes 1, 2 and 3 are active antifungal agents. Minimal inhibitory concentrations (MIC) were in the range of 0.25-4.68 microg/ml for all tested fungal strains. Considerably larger differences were found for minimal fungicidal concentrations (MFC). In the case of yeast-like fungi, the fungicidal effect was generally observed at organotin compounds concentrations of 2.34-9.37 microg/ml. The MFC values for filamentous fungi were considerably higher and were in the range of 18.74-50 microg/ml. In conclusion, organotin compounds 1, 2 and 3 showed high fungistatic and fungicidal activities against different species of pathogenic and nonpathogenic fungi. However, they were also highly cytotoxic towards two mammalian cell lines. PMID:19688632

  4. Antibacterial, antifungal and in vitro antileukaemia activity of metal complexes with thiosemicarbazones

    PubMed Central

    Pahontu, Elena; Julea, Felicia; Rosu, Tudor; Purcarea, Victor; Chumakov, Yurie; Petrenco, Petru; Gulea, Aurelian

    2015-01-01

    1-phenyl-3-methyl-4-benzoyl-5-pyrazolone 4-ethyl-thiosemicarbazone (HL) and its copper(II), vanadium(V) and nickel(II) complexes: [Cu(L)(Cl)]·C2H5OH·(1), [Cu(L)2]·H2O (2), [Cu(L)(Br)]·H2O·CH3OH (3), [Cu(L)(NO3)]·2C2H5OH (4), [VO2(L)]·2H2O (5), [Ni(L)2]·H2O (6), were synthesized and characterized. The ligand has been characterized by elemental analyses, IR, 1H NMR and 13C NMR spectroscopy. The tridentate nature of the ligand is evident from the IR spectra. The copper(II), vanadium(V) and nickel(II) complexes have been characterized by different physico-chemical techniques such as molar conductivity, magnetic susceptibility measurements and electronic, infrared and electron paramagnetic resonance spectral studies. The structures of the ligand and its copper(II) (2, 4), and vanadium(V) (5) complexes have been determined by single-crystal X-ray diffraction. The composition of the coordination polyhedron of the central atom in 2, 4 and 5 is different. The tetrahedral coordination geometry of Cu was found in complex 2 while in complex 4, it is square planar, in complex 5 the coordination polyhedron of the central ion is distorted square pyramid. The in vitro antibacterial activity of the complexes against Escherichia coli, Salmonella abony, Staphylococcus aureus, Bacillus cereus and the antifungal activity against Candida albicans strains was higher for the metal complexes than for free ligand. The effect of the free ligand and its metal complexes on the proliferation of HL-60 cells was tested. PMID:25708540

  5. Artificial oocyte activation in intracytoplasmic sperm injection cycles using testicular sperm in human in vitro fertilization

    PubMed Central

    Kang, Hee Jung; Lee, Sun-Hee; Park, Yong-Seog; Lim, Chun Kyu; Ko, Duck Sung; Yang, Kwang Moon

    2015-01-01

    Objective Artificial oocyte activation (AOA) is an effective method to avoid total fertilization failure in human in vitro fertilization-embryo transfer (IVF-ET) cycles. AOA performed using a calcium ionophore can induce calcium oscillation in oocytes and initiate the fertilization process. We evaluated the usefulness of AOA with a calcium ionophore in cases of total fertilization failure in previous cycles and in cases of severe male factor infertility patients with non-motile spermatozoa after pentoxifylline (PF) treatment. Methods The present study describes 29 intracytoplasmic sperm injection (ICSI)-AOA cycles involving male factor infertility at Cheil General Hospital from January 2006 to June 2013. Patients were divided into two groups (control, n=480; AOA, n=29) depending on whether or not AOA using a calcium ionophore (A23187) was performed after testicular sperm extraction-ICSI (TESE-ICSI). The AOA group was further split into subgroups according to sperm motility after PF treatment: i.e., motile sperm-injected (n=12) and non-motile sperm-injected (n=17) groups (total n=29 cycles). Results The good embryo rate (52.3% vs. 66.9%), pregnancy rate (20.7% vs. 52.1%), and delivery rate (10.3% vs. 40.8%) were lower in the PF/AOA group than in the control group. When evaluating the effects of restoration of sperm motility after PF treatment on clinical outcomes there was no difference in fertilization rate (66.6% vs. 64.7% in non-motile and motile sperm, respectively), pregnancy rate (17.6% vs. 33.3%), or delivery rate (5.9% vs. 16.7%) between the two groups. Conclusion We suggest that oocyte activation is a useful method to ensure fertilization in TESE-ICSI cycles regardless of restoration of sperm motility after PF treatment. AOA may be useful in selected patients who have a low fertilization rate or total fertilization failure. PMID:26161332

  6. Enhancement in intramolecular interactions and in vitro biological activity of a tripodal tetradentate system upon complexation.

    PubMed

    Tyagi, Nidhi; Viji, Mambattakkara; Karunakaran, Suneesh C; Varughese, Sunil; Ganesan, Shilpa; Priya, Sulochana; Saneesh Babu, P S; Nair, Asha S; Ramaiah, Danaboyina

    2015-09-21

    Novel biomimetic mononuclear complexes, [Fe()Cl2](+) () and [Cu()(H2O)](2+) () based on naphthalimide appended tripodal tetradentate ligand ( = 2,2',2''-(3,3',3''-(2,2',2''-nitrilotris(methylene)tris(1H-benzo[d]imidazole-2,1-diyl))tris(propane-3,1-diyl))tris(1H-benzo-[de]isoquinoline-1,3(2H)-dione)) have been synthesized and characterized by various analytical and spectral techniques. In addition, the structures of the ligand () and complex were established unambiguously through X-ray crystal structure analysis. Uniquely, the coordination with a metal ion modified the ligand scaffold to interact efficiently with ct-DNA (groove binding) as well as protein (hydrophobic and/or electrostatic interactions). We have determined the affinity of these complexes for DNA/protein and the values are found to be in the range, KDNA = 0.34-1.01 × 10(4) M(-1) and KBSA = 4.1-5.0 × 10(5) M(-1). Furthermore, the fluorescence quenching of BSA with complexes and occurs through a static mechanism and affects the conformation of BSA around the tryptophan residues. The in vitro biological studies of these systems employing HeLa cell lines indicated that both these complexes exhibited enhanced cytotoxicity (IC50 = 32 ± 0.19 and 10 ± 0.21 ?M for complexes and , respectively), when compared to the ligand () (IC50 = 150 ?M). Interestingly, both the complexes ( and ) were found to be non-toxic to normal H9C2 cell lines. The mechanism of in vitro biological activity of these complexes has been evaluated through a variety of techniques: acridine orange/ethidium bromide, DAPI staining studies, annexin V-FITC/PI and poly(ADP-ribose)-polymerase (PARP) cleavage, which confirmed the apoptotic mediated cell death. Our results demonstrate the importance of complexation of the naphthalimide ligand () as well as the potential of these biomimetic metal complexes as cytotoxic and anticancer agents. PMID:26244289

  7. The in vitro and ex vivo antioxidant properties, and hypolipidemic activity of CGP 2881.

    PubMed

    Feldman, D L; Mogelesky, T C; Sharif, R; Sawyer, W K; Jeune, M; Hu, C W; Leonards, K S; Prescott, M F

    1999-06-01

    This report describes the in vitro and ex vivo antioxidant properties of a new antioxidant, CGP 2881. This compound is structurally similar to probucol, in that both compounds contain bis-tertiary butyl phenyl groups. However, CGP 2881 consistently inhibited CuSO4 (Cu2+)- and macrophage (MO)-induced oxidation of human low density lipoproteins (LDL) more potently than equimolar concentrations of probucol. CGP 2881 (1 mumol/l) prolonged the lag phase of diene formation during Cu(2+)-induced LDL oxidation by 3.4 versus 1.5-fold prolongation with 1 mumol/l probucol (P < 0.05 vs CGP 2881). The IC50 for inhibiting the formation of Cu(2+)-induced thiobarbituric acid-reactive substances (TBARS) was 0.15 mumol/l for CGP 2881, versus approximately 10 mumol/l for probucol. The IC50 for MO-induced oxidation of LDL (TBARS) was 0.64 mumol/l. In contrast, 1 mumol/l probucol failed to inhibit MO-induced oxidation of LDL. Treatment of cholic acid/cholesterol-fed rats with CGP 2881 (50 mg/kg per day, orally for 5 days) inhibited ex vivo Cu(2+)-induced oxidation (TBARS) of the very low density lipoproteins (VLDL) + LDL lipoprotein fraction by 93% versus vehicle controls (P < 0.0001), and prolonged the lag phase for Cu(2+)-induced diene formation by 3.4-fold over vehicle-treated controls. Five days of orally administered CGP 2881 reduced plasma total cholesterol and LDL cholesterol levels to 55 and 54% of vehicle-treated controls, respectively (P < 0.05). In contrast, probucol had no appreciable effect on plasma total cholesterol or LDL cholesterol levels, unless administered for longer than 5 days. Treatment of hypercholesterolemic rabbits with 50 mg/kg per day orally for 5-12 days delayed the lag phase of diene formation during LDL oxidation by 4.3-fold over controls. However, the relative antioxidant potencies of CGP 2881 and probucol seen with oral administration to hypercholesterolemic rabbits were reversed when the compounds were given intravenously. In addition, the effects of these antioxidants were potentiated when given to normocholesterolemic rabbits compared to hypercholesterolemic animals. These data establish that CGP 2881 demonstrates hypolipidemic activity and is a substantially more potent antioxidant than probucol (in vitro and ex vivo). CGP 2881 may be useful as a new antioxidant tool in the effort to better understand the atherogenicity of oxidized LDL (oxLDL). PMID:10407495

  8. In vitro inhibition effect and structure-activity relationships of some saccharin derivatives on erythrocyte carbonic anhydrase I and II.

    PubMed

    Sonmez, Fatih; Bilen, Cigdem; Sumersan, Sinem; Gencer, Nahit; Isik, Semra; Arslan, Oktay; Kucukislamoglu, Mustafa

    2014-02-01

    In this study, in vitro inhibitory effects of some saccharin derivatives on purified carbonic anhydrase I and II were investigated using CO2 as a substrate. The results showed that all compounds inhibited the hCA I and hCA II enzyme activities. Among the compounds, 6-(p-tolylthiourenyl) saccharin (6m) was found to be the most active one for hCA I activity (IC50=13.67 ?M) and 6-(m-methoxyphenylurenyl) saccharin (6b) was found to be the most active one for hCA II activity (IC50=6.54 ?M). Structure-activity relationships (SARs) study showed that, generally, thiourea derivatives (6l--v) inhibited more hCA I and hCA II than urea derivatives (6a-k). All compounds (excluding 6c and 6r) have higher inhibitory activity on hCA II than on hCA I. PMID:23339426

  9. In Vitro inhibitory activity of Alpinia katsumadai extracts against influenza virus infection and hemagglutination

    PubMed Central

    2010-01-01

    Background Alpinia katsumadai (AK) extracts and fractions were tested for in vitro antiviral activities against influenza virus type A, specially human A/PR/8/34 (H1N1) and avian A/Chicken/Korea/MS96/96 (H9N2), by means of time-of-addition experiments; pre-treatment, simultaneous treatment, and post treatment. Results In pre-treatment assay, the AK extracts and AK fractions did not show significant antiviral activity. During the simultaneous treatment assay, one AK extract and five AK fractions designated as AK-1 to AK-3, AK-5, AK-10, and AK-11 showed complete inhibition of virus infectivity against A/PR/8/34 (H1N1) and A/Chicken/Korea/MS96/96 (H9N2). The 50% effective inhibitory concentrations (EC50) of these one AK extracts and five AK fractions with exception of the AK-9 were from 0.8 ± 1.4 to 16.4 ± 4.5 ?g/mL against A/PR/8/34 (H1N1). The two AK extracts and three AK fractions had EC50 values ranging from <0.39 ± 0.4 to 2.3 ± 3.6 ?g/mL against A/Chicken/Korea/MS96/96 (H9N2). By the hemagglutination inhibition (HI) assay, the two AK extracts and five AK fractions completely inhibited viral adsorption onto chicken RBCs at less than 100 ?g/mL against both A/PR/8/34 (H1N1) and A/Chicken/Korea/MS96/96 (H9N2). Interestingly, only AK-3 was found with inhibition for both viral attachment and viral replication after showing extended antiviral activity during the post treatment assay and quantitative real-time PCR. Conclusions These results suggest that AK extracts and fractions had strong anti-influenza virus activity that can inhibit viral attachment and/or viral replication, and may be used as viral prophylaxis. PMID:21062499

  10. Using in vitro structural alerts for chromosome damage to predict in vivo activity and direct future testing.

    PubMed

    Canipa, Steven; Cayley, Alex; Drewe, William C; Williams, Richard V; Hamada, Shuichi; Hirose, Akihiko; Honma, Masamitsu; Morita, Takeshi

    2016-01-01

    While the in vivo genotoxicity of a compound may not always correlate well with its activity in in vitro test systems, for certain compound classes a good overlap may exist between the two endpoints. The difficulty, however, lies in establishing the cases where this relationship holds true and selecting the most appropriate protocol to highlight any potential in vivo hazard. With this in mind, a project was initiated in which existing structural alerts for in vitro chromosome damage in the expert system Derek Nexus were assessed for their relevance to in vivo activity by assessing their predictivity against an in vivo chromosome damage data set. An expert assessment was then made of selected alerts. Information regarding the findings from specific in vivo tests was added to the alert along with any significant correlations between activity and test protocol or mechanism. A total of 32 in vitro alerts were updated using this method resulting in a significant improvement in the coverage of in vivo chromosome damage in Derek Nexus against a data set compiled by the mammalian mutagenicity study group of Japan. The detailed information relating to in vivo activity and protocol added to the alerts in combination with the mechanistic information provided will prove useful in directing the further testing of compounds of interest. PMID:26142242

  11. In vitro antiplasmodial, antileishmanial and antitrypanosomal activities of selected medicinal plants used in the traditional Arabian Peninsular region

    PubMed Central

    2012-01-01

    Background Worldwide particularly in developing countries, a large proportion of the population is at risk for tropical parasitic diseases. Several medicinal plants are still used traditionally against protozoal infections in Yemen and Saudi Arabia. Thus the present study investigated the in vitro antiprotozoal activity of twenty-five plants collected from the Arabian Peninsula. Methods Plant materials were extracted with methanol and screened in vitro against erythrocytic schizonts of Plasmodium falciparum, intracellular amastigotes of Leishmania infantum and Trypanosoma cruzi and free trypomastigotes of T. brucei. Cytotoxic activity was determined against MRC-5 cells to assess selectivity. The criterion for activity was an IC50?4. Results Antiplasmodial activity was found in the extracts of Chrozophora oblongifolia, Ficus ingens, Lavandula dentata and Plectranthus barbatus. Amastigotes of T. cruzi were affected by Grewia erythraea, L. dentata, Tagetes minuta and Vernonia leopoldii. Activity against T. brucei was obtained in G. erythraea, L. dentata, P. barbatus and T. minuta. No relevant activity was found against L. infantum. High levels of cytotoxicity (MRC-5 IC50?activities were noted in Cupressus sempervirens, Kanahia laniflora and Kniphofia sumarae. Conclusion The results endorse that medicinal plants can be promising sources of natural products with antiprotozoal activity potential. The results support to some extent the traditional uses of some plants for the treatment of parasitic protozoal diseases. PMID:22520595

  12. In vitro antioxidant and inhibitory activity of water decoctions of carob tree (Ceratonia siliqua L.) on cholinesterases, ?-amylase and ?-glucosidase.

    PubMed

    Custódio, Luísa; Patarra, Joăo; Alberício, Fernando; Neng, Nuno Rosa; Nogueira, José Manuel Floręncio; Romano, Anabela

    2015-01-01

    This work reports the in vitro inhibitory activity of water decoctions of leaves, germ flour, pulp, locust bean gum and stem bark of carob tree on ?-amylase, ?-glucosidase, acetylcholinesterase and butyrylcholinesterase. The antioxidant activity and the chemical characterisation of the extracts made by spectrophotometric assays and by high-performance liquid chromatography are also reported. Leaves and stem bark decoctions strongly inhibited all the enzymes tested, had significant antioxidant activity and the highest total phenolics content. The major compounds were identified as gallic acid in the leaves and gentisic acid in the stem bark. PMID:25582851

  13. In vitro germination characteristics of maize pollen to detect biological activity of environmental pollutants

    SciTech Connect

    Pfahler, P.L.

    1981-01-01

    In vitro pollen germination in maize was evaluated as a method of assessing the mutagenic and physiological effects of environmental pollutants on higher organisms. The extent of mutations was effectively determined by testing sporophytes homozygous for the mutated allele. Physiological effects were effectively determined when chemical agents were added directly into the in vitro medium. Exposure of pollen grains during in vitro germination to ultraviolet radiation in the range 280-320 nm produced little or no change in the germination or rupture percentage but a sharp decrease in pollen tube growth after 1 hr.

  14. In Vitro Activity of Isavuconazole and Comparators against Clinical Isolates of the Mucorales Order

    PubMed Central

    Jensen, Rasmus Hare; Meletiadis, Joseph

    2015-01-01

    The in vitro activity of isavuconazole against Mucorales isolates measured by EUCAST E.Def 9.2 and CLSI M38-A2 methodologies was investigated in comparison with those of amphotericin B, posaconazole, and voriconazole. Seventy-two isolates were included: 12 of Lichtheimia corymbifera, 5 of Lichtheimia ramosa, 5 of group I and 9 of group II of Mucor circinelloides, 9 of Rhizomucor pusillus, 26 of Rhizopus microsporus, and 6 of Rhizopus oryzae. Species identification was confirmed by internal transcribed spacer (ITS) sequencing. EUCAST MICs were read on day 1 (EUCAST-d1) and day 2 (EUCAST-d2), and CLSI MICs were read on day 2 (CLSI-d2). Isavuconazole MIC50s (range) (mg/liter) by EUCAST-d1, CLSI-d2, and EUCAST-d2 were 1 (0.125 to 16), 1 (0.125 to 2), and 4 (0.5 to >16), respectively, across all isolates. The similar values for comparator drugs were as follows: posaconazole, 0.25 (?0.03 to >16), 0.25 (0.06 to >16), and 1 (0.06 to >16); amphotericin, 0.06 (?0.03 to 0.5), 0.06 (?0.03 to 0.25), and 0.125 (?0.03 to 1); voriconazole, 16 (2 to >16), 8 (1 to >16), and >16 (8 to >16), respectively. Isavuconazole activity varied by species: Lichtheimia corymbifera, 1 (0.5 to 2), 1 (1 to 2), and 2 (1 to 4); Lichtheimia ramosa, 0.25 (0.125 to 0.5), 1 (0.5 to 2), and 2 (0.5 to 4); Rhizomucor pusillus, 0.5 (0.5 to 1), 1 (0.125 to 1), and 2 (1 to 2); Rhizopus microsporus, 1 (0.5 to 4), 0.5 (0.125 to 1), and 4 (1 to 8); and Rhizopus oryzae, 1 (0.5 to 4), 1 (0.125 to 2), and 4 (0.5 to 8), respectively, were more susceptible than Mucor circinelloides: group I, 8 (4 to 8), 4 (2 to 4), and 16 (2 to 16), respectively, and group II, 8 (1 to 16), 8 (1 to 8), and 16 (4 to >16), respectively. This was also observed for posaconazole. The essential agreement was best between EUCAST-d1 and CLSI-d2 (75% to 83%). Isavuconazole displayed in vitro activity against Mucorales isolates with the exception of Mucor circinelloides. The MICs were in general 1 to 3 steps higher than those for posaconazole. However, in the clinical setting this may be compensated for by the higher exposure at standard dosing. PMID:26438494

  15. In Vitro Activity of Isavuconazole and Comparators against Clinical Isolates of the Mucorales Order.

    PubMed

    Arendrup, Maiken Cavling; Jensen, Rasmus Hare; Meletiadis, Joseph

    2015-12-01

    The in vitro activity of isavuconazole against Mucorales isolates measured by EUCAST E.Def 9.2 and CLSI M38-A2 methodologies was investigated in comparison with those of amphotericin B, posaconazole, and voriconazole. Seventy-two isolates were included: 12 of Lichtheimia corymbifera, 5 of Lichtheimia ramosa, 5 of group I and 9 of group II of Mucor circinelloides, 9 of Rhizomucor pusillus, 26 of Rhizopus microsporus, and 6 of Rhizopus oryzae. Species identification was confirmed by internal transcribed spacer (ITS) sequencing. EUCAST MICs were read on day 1 (EUCAST-d1) and day 2 (EUCAST-d2), and CLSI MICs were read on day 2 (CLSI-d2). Isavuconazole MIC50s (range) (mg/liter) by EUCAST-d1, CLSI-d2, and EUCAST-d2 were 1 (0.125 to 16), 1 (0.125 to 2), and 4 (0.5 to >16), respectively, across all isolates. The similar values for comparator drugs were as follows: posaconazole, 0.25 (?0.03 to >16), 0.25 (0.06 to >16), and 1 (0.06 to >16); amphotericin, 0.06 (?0.03 to 0.5), 0.06 (?0.03 to 0.25), and 0.125 (?0.03 to 1); voriconazole, 16 (2 to >16), 8 (1 to >16), and >16 (8 to >16), respectively. Isavuconazole activity varied by species: Lichtheimia corymbifera, 1 (0.5 to 2), 1 (1 to 2), and 2 (1 to 4); Lichtheimia ramosa, 0.25 (0.125 to 0.5), 1 (0.5 to 2), and 2 (0.5 to 4); Rhizomucor pusillus, 0.5 (0.5 to 1), 1 (0.125 to 1), and 2 (1 to 2); Rhizopus microsporus, 1 (0.5 to 4), 0.5 (0.125 to 1), and 4 (1 to 8); and Rhizopus oryzae, 1 (0.5 to 4), 1 (0.125 to 2), and 4 (0.5 to 8), respectively, were more susceptible than Mucor circinelloides: group I, 8 (4 to 8), 4 (2 to 4), and 16 (2 to 16), respectively, and group II, 8 (1 to 16), 8 (1 to 8), and 16 (4 to >16), respectively. This was also observed for posaconazole. The essential agreement was best between EUCAST-d1 and CLSI-d2 (75% to 83%). Isavuconazole displayed in vitro activity against Mucorales isolates with the exception of Mucor circinelloides. The MICs were in general 1 to 3 steps higher than those for posaconazole. However, in the clinical setting this may be compensated for by the higher exposure at standard dosing. PMID:26438494

  16. Variations in cercarial production and the level of in vitro activation of metacercariae of two different isolates of Fasciola hepatica.

    PubMed

    Januszkiewicz, Kamil; Norbury, Luke J; Wilkowski, Przemys?aw; Zawistowska-Deniziak, Anna; Weso?owska, Agnieszka; Wedrychowicz, Halina

    2015-09-01

    Fasciola hepatica infections cause large economic losses and are a serious veterinary medicine problem in many regions of the world. Recent studies examining fascioliasis in the bison population from Bialowieza National Park have shown that the prevalence of infection with this parasite is up to 100%. Liver flukes isolated from bison from Bialowieza National Park in Poland were compared with a fluke strain originally obtained from the Central Veterinary Laboratory, Weybridge, UK, to determine variations in cercarial production and establish the ability of their metacercariae to activate in vitro. Some small differences in cercarial production between the two isolates are shown, while significant differences in the ability of their metacercariae to activate in vitro were observed. PMID:26204191

  17. Tauroursodeoxycholic acid prevents stress induced aggregation of proteins in vitro and promotes PERK activation in HepG2 cells.

    PubMed

    Gani, Amina R; Uppala, Jagadeesh Kumar; Ramaiah, Kolluru V A

    2015-02-15

    Tauroursodeoxycholic acid (TUDCA) a bile salt and chemical chaperone reduces stress-induced aggregation of proteins; activates PERK [PKR (RNA-dependent protein kinase)-like ER (endoplasmic reticulum) kinase] or EIF2AK3, one of the hall marks of ER stress induced unfolded protein response (UPR) in human hepatoblastoma HepG2 cells; prevents heat and dithiothreitol (DTT) induced aggregation of BSA (bovine serum albumin), and reduces ANS (1-anilino-naphthalene-8-sulfonate) bound BSA fluorescence in vitro. TUDCA inactivates heat treated, but not the native EcoR1 enzyme, and reduces heat-induced aggregation and activity of COX-1 (cyclooxygenase enzyme-1) in vitro. These findings suggest that TUDCA binds to the hydrophobic regions of proteins and prevents their subsequent aggregation. This may stabilize unfolded proteins that can mount UPR or facilitate their degradation through cellular degradation pathways. PMID:25579883

  18. Alternatively activated macrophages actively inhibit proliferation of peripheral blood lymphocytes and CD4+ T cells in vitro.

    PubMed Central

    Schebesch, C; Kodelja, V; Müller, C; Hakij, N; Bisson, S; Orfanos, C E; Goerdt, S

    1997-01-01

    We compared the immunological functions of interferon-gamma (IFN-gamma)-induced, classically activated macrophages (caM phi) and of interleukin-4 (IL-4)- and glucocorticoid-induced, alternatively activated macrophages (aaM phi) in a human co-culture system in vitro. Proliferation of peripheral blood leucocytes (PBL) or CD4+ T cells mediated by optimal doses of phytohaemagglutinin (PHA) or concanavalin A (Con A) was only marginally influenced by caM phi, but was strongly inhibited by aaM phi. The degree of lymphocyte proliferation sustained in the presence of caM phi was gradually reduced in a dose-dependent fashion by the addition of aaM phi. Flow cytometric analysis revealed that expression of costimulatory molecules such as CD11a, CD40, CD54, CD58, CD80 and CD86 did not vary significantly between caM phi and aaM phi and was low for CD58, CD80 and CD86. As shown by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, IL-10 was expressed in caM phi, aaM phi and control macrophages; the level of expression of IL-10 was slightly enhanced in aaM phi. Neither neutralizing anti-IL-10 antibodies, indomethacin nor NG-monomethyl-L-arginine (NMMLA) was able to reverse aaM phi-mediated inhibition of lymphocyte proliferation. Of several agents interfering with various second messenger pathways, cAMP and the Ca(2+)-ionophore A23187 inhibited differentiation of cultured human monocytes into phenotypically mature aaM phi expressing MS-1 high molecular weight protein (MS-1-HMWP) and RM 3/1 antigen, and prevented the suppressive action of aaM phi on lymphocyte proliferation. In conclusion, these results who that aaM phi actively inhibit mitogen-mediated proliferation of PBL and CD4+ T cells independently of the expression of costimulatory molecules and of IL-10, NO or prostaglandin synthesis, and that inhibition of phenotypic differentiation of aaM phi is paralleled by a lack of functional maturation. Thus, fully matured aaM phi may be functional in down-regulating CD4+ T-cell-mediated immune reactions by an as yet unknown mechanism. PMID:9497489

  19. EFFECT OF WATER POLLUTANTS AND OTHER CHEMICALS UPON RIBONUCLEASE ACTIVITY IN VITRO

    EPA Science Inventory

    Ribonuclease was treated in vitro with 73 chemicals, many of which are environmental pollutants, including inorganic, organic, and metal-organic chemicals, pesticides, and other biocides, alkyl and aryl industrial pollutants, and certain additional chemicals, to determine their e...

  20. Anticancer activity of peach and plum extracts against human breast cancer in vitro and in vivo 

    E-print Network

    Noratto Dongo, Giuliana Doris

    2009-05-15

    "Black Splendor" (BS) on tumor breast cells in vitro and in vivo, to elucidate the molecular mechanisms behind the cancer growth-suppression of the phenolics identified in peach and plum extracts for their chemopreventive potential and to evaluate...

  1. Comparative in vitro and in vivo antimalarial activity of the indole alkaloids ellipticine, olivacine, cryptolepine and a synthetic cryptolepine analog.

    PubMed

    Rocha e Silva, L F; Montoia, A; Amorim, R C N; Melo, M R; Henrique, M C; Nunomura, S M; Costa, M R F; Andrade Neto, V F; Costa, D S; Dantas, G; Lavrado, J; Moreira, R; Paulo, A; Pinto, A C; Tadei, W P; Zacardi, R S; Eberlin, M N; Pohlit, A M

    2012-12-15

    Indole alkaloids ellipticine (1), cryptolepine triflate (2a), rationally designed 11-(4-piperidinamino)cryptolepine hydrogen dichloride (2b) and olivacine (3) (an isomer of 1) were evaluated in vitro against Plasmodium falciparum and in vivo in Plasmodium berghei-infected mice. 1-3 inhibited P. falciparum (IC???1.4 ?M, order of activity: 2b>1>2a>3). In vitro toxicity to murine macrophages was evaluated and revealed selectivity indices (SI) of 10-12 for 2a and SI>2.8×10˛ for 1, 2b and 3. 1 administered orally at 50mg/kg/day was highly active against P. berghei (in vivo inhibition compared to untreated control (IVI)=100%, mean survival time (MST)>40 days, comparable activity to chloroquine control). 1 administered orally and subcutaneously was active at 10 mg/kg/day (IVI=70-77%; MST=27-29 days). 3 exhibited high oral activity at ?50 mg/kg/day (IVI=90-97%, MST=23-27 days). Cryptolepine (2a) administered orally and subcutaneously exhibited moderate activity at 50mg/kg/day (IVI=43-63%, MST=24-25 days). At 50 mg/kg/day, 2b administered subcutaneously was lethal to infected mice (MST=3 days) and moderately active when administered orally (IVI=45-55%, MST=25 days). 1 and 3 are promising compounds for development of antimalarials. PMID:23092722

  2. A Novel Antifungal Is Active against Candida albicans Biofilms and Inhibits Mutagenic Acetaldehyde Production In Vitro

    PubMed Central

    Nieminen, Mikko T.; Novak-Frazer, Lily; Rautemaa, Vilma; Rajendran, Ranjith; Sorsa, Timo; Ramage, Gordon; Bowyer, Paul; Rautemaa, Riina

    2014-01-01

    The ability of C. albicans to form biofilms is a major virulence factor and a challenge for management. This is evident in biofilm-associated chronic oral-oesophageal candidosis, which has been shown to be potentially carcinogenic in vivo. We have previously shown that most Candida spp. can produce significant levels of mutagenic acetaldehyde (ACH). ACH is also an important mediator of candidal biofilm formation. We have also reported that D,L-2-hydroxyisocaproic acid (HICA) significantly inhibits planktonic growth of C. albicans. The aim of the present study was to investigate the effect of HICA on C. albicans biofilm formation and ACH production in vitro. Inhibition of biofilm formation by HICA, analogous control compounds or caspofungin was measured using XTT to measure biofilm metabolic activity and PicoGreen as a marker of biomass. Biofilms were visualised by scanning electron microscopy (SEM). ACH levels were measured by gas chromatography. Transcriptional changes in the genes involved in ACH metabolism were measured using RT-qPCR. The mean metabolic activity and biomass of all pre-grown (4, 24, 48 h) biofilms were significantly reduced after exposure to HICA (p<0.05) with the largest reductions seen at acidic pH. Caspofungin was mainly active against biofilms pre-grown for 4 h at neutral pH. Mutagenic levels (>40 µM) of ACH were detected in 24 and 48 h biofilms at both pHs. Interestingly, no ACH production was detected from D-glucose in the presence of HICA at acidic pH (p<0.05). Expression of genes responsible for ACH catabolism was up-regulated by HICA but down-regulated by caspofungin. SEM showed aberrant hyphae and collapsed hyphal structures during incubation with HICA at acidic pH. We conclude that HICA has potential as an antifungal agent with ability to inhibit C. albicans cell growth and biofilm formation. HICA also significantly reduces the mutagenic potential of C. albicans biofilms, which may be important when treating bacterial-fungal biofilm infections. PMID:24867320

  3. A Select Combination of Clinically Relevant Phytoestrogens Enhances Estrogen Receptor ?-Binding Selectivity and Neuroprotective Activities in Vitro and in Vivo

    E-print Network

    Zhao, Liqin; Mao, Zisu; Brinton, Roberta Diaz

    2009-01-01

    of Clinically Relevant Phytoestrogens Enhances Estrogen Receptor ?-Binding Selectivity and Neuroprotective Activities in Vitro and in Vivo Liqin Zhao, Zisu Mao, and Roberta Diaz Brinton Department of Pharmacology and Pharmaceutical Sciences (L.Z., Z.M., R.D.B....), School of Pharmacy, and Neuroscience Program (R.D.B.), University of Southern California, Los Angeles, California 90089 We have previously shown that a number of naturally occurring phytoestrogens and derivatives were effective to induce some measures...

  4. In vitro antimycotic activity and nail permeation models of a piroctone olamine (octopirox) containing transungual water soluble technology.

    PubMed

    Dubini, Francesco; Bellotti, Maria Grazia; Frangi, Alessandra; Monti, Daniela; Saccomani, Luigi

    2005-01-01

    Several in vitro studies with a new medical device (Myfungar) containing 0.5% of piroctone olamine (CAS 68890-66-4, octopirox) in a hydroxypropyl chitosan hydroalcoholic solution were performed. The chemical name of the active ingredient is 1-hydroxy-4-methyl-6 (2,4,4-trimethylpentyl)-2(1H)-pyridone; combination with 2-amino-ethanol (1:1). The antimycotic activity was determined in the most common fungi responsible of nail infections such as Candida parapsilosis, Scopulariopsis brevicaulis or Trichophyton rubrum. The minimum inhibitory concentration (MIC), found by means of the broth dilution susceptibility method, ranged between 0.0003% and 0.006% for all pathogens considered. The in vitro permeation study was performed by using bovine hoof membranes inserted in a modified Gummer vertical permeation cell. The experiment showed, at 30 h, a retention of the product in the nail membranes by 11.1% of the applied dose. No piroctone permeation through bovine hoof membranes could be detected by HPLC due to the limit of quantitation of this method. On the other hand, permeation of nail membranes has been demonstrated by a biological assay: the study of in vitro permeation through bovine hoof membranes, performed by biological assay, showed dose-dependent inhibition rings of T. rubrum growth by the tested device, placed either on disks for antibiogram or on nail fragments. The placebo did not show any inhibition. In vitro experimental infection by T. rubrum showed a preventive activity of the tested device on fungal growth as well as a curative activity, as the pathogen was eradicated by the tested solution in previously infected nails. PMID:16149717

  5. Influence of age on in vitro effect of cadmium on rat liver cytochrome P-450 concentration and monooxygenases activity.

    PubMed

    Jahn, F; Klinger, W

    1982-02-01

    Cadmium++ added in vitro destroys rat liver cytochrome P-450 (cyt. P-450) with increasing age by 25-50%. Ethylmorphine N-demethylation is inhibited only in rats 30-days old and thereafter. Ethoxycoumarin 0-deethylation is inhibited even in newborn rats, and the maximal inhibition appears to increase with age. It is concluded that in all age groups cadmium resistant cyt. P-450 subspecies are present. Ethoxycoumarin 0-deethylase activity possibly indicates the cadmium sensitive P-450 fraction. PMID:6978593

  6. The in vivo and in vitro effects of caffeine on rat immune cells activities: B, T and NK cells.

    PubMed

    Kantamala, D; Vongsakul, M; Satayavivad, J

    1990-12-01

    The effect of caffeine (naturally occurring plant methylxanthine) on immunological cell activities in Sprague-Dawley rat both in vivo and in vitro was studied. A cytotoxic assay was done to study natural killer (NK) cells and a proliferation assay was performed for T and B cell activities. Three different doses of caffeine i.e., 2, 6 and 18 mg/kg/day were administered chronically to Sprague-Dawley rats to assess the effects in vivo. Both NK cell cytotoxicity and B cell proliferative response to pokeweed mitogen (PWM) showed significant decreases (P less than 0.05) in rats treated with 6 mg/kg/day, whereas the T cell proliferative response to phytohemagglutinin-P (PHA-P) was significantly increased (P less than 0.05) in the rats treated with 18 mg/kg/day. In vitro, caffeine significantly decreases (P less than 0.05) B and T cell proliferative responses to PWM and PHA-P at added caffeine concentrations of 10, 20 and 40 micrograms/ml. However, no effect was observed on NK cells activity. Furthermore, in vitro, a broader dose range of caffeine (1, 10, 100 and 1,000 micrograms/ml) exhibited dose-dependent inhibition of both B and T cell proliferative responses. PMID:2091662

  7. Toxocara canis: potential activity of natural products against second-stage larvae in vitro and in vivo.

    PubMed

    Reis, Mariana; Trinca, Alcione; Ferreira, Maria José U; Monsalve-Puello, Ana R; Grácio, Maria Amélia A

    2010-10-01

    The anthelmintic activity of extracts from Chenopodiumambrosioides, Pycnanthusangolensis and Nutridesintox was in vitro and in vivo investigated, against Toxocaracanis larvae. The in vitro assays results showed that the aqueous extract of Nutridesintox was the most effective, followed by C. ambrosioides extracts, hexane, dichloromethane and the infusion. P. angolensis extracts showed a lower anthelmintic activity compared to the other natural products. For the in vivo assays, Nutridesintox, the hexane extract and the infusion of C. ambrosioides were administered orally to T. canis-infected mice, in single doses, during three consecutive days. The efficacy was evaluated on the 17th day post-infection, not only by counting T. canis larvae in the tissues but also by ELISA detection of IgM and IgG antibodies and histological analysis of liver and lungs. The different treatments did not reduce the larvae burden and had no influence on the antibodies dynamic. Interestingly, a reduction on the inflammatory infiltrates was observed in the liver and lung sections of the group treated with the hexane extract of C. ambrosioides. In conclusion, the hexane extract of C. ambrosioides is of further research interest, as it showed an anthelmintic activity in vitro and a reduction on the inflammatory reaction produced by the infection of T. canis larvae in vivo. PMID:20447397

  8. Buparvaquone is active against Neospora caninum in vitro and in experimentally infected mice

    PubMed Central

    Müller, Joachim; Aguado-Martinez, Adriana; Manser, Vera; Balmer, Vreni; Winzer, Pablo; Ritler, Dominic; Hostettler, Isabel; Arranz-Solís, David; Ortega-Mora, Luis; Hemphill, Andrew

    2015-01-01

    The naphthoquinone buparvaquone is currently the only drug used against theileriosis. Here, the effects of buparvaquone were investigated in vitro and in an experimental mouse model for Neospora caninum infection. In 4-day proliferation assays, buparvaquone efficiently inhibited N.?caninum tachyzoite replication (IC50?=?4.9?nM; IC100?=?100?nM). However, in the long term tachyzoites adapted and resumed proliferation in the presence of 100?nM buparvaquone after 20 days of cultivation. Parasiticidal activity was noted after 9 days of culture in 0.5?µM or 6 days in 1?µM buparvaquone. TEM of N.?caninum infected fibroblasts treated with 1?µM buparvaquone showed that the drug acted rather slowly, and ultrastructural changes were evident only after 3–5 days of treatment, including severe alterations in the parasite cytoplasm, changes in the composition of the parasitophorous vacuole matrix and a diminished integrity of the vacuole membrane. Treatment of N.?caninum infected mice with buparvaquone (100?mg/kg) either by intraperitoneal injection or gavage prevented neosporosis symptoms in 4 out of 6 mice in the intraperitoneally treated group, and in 6 out of 7 mice in the group receiving oral treatment. In the corresponding controls, all 6 mice injected intraperitoneally with corn oil alone died of acute neosporosis, and 4 out of 6 mice died in the orally treated control group. Assessment of infection intensities in the treatment groups showed that, compared to the drug treated groups, the controls showed a significantly higher parasite load in the lungs while cerebral parasite load was higher in the buparvaquone-treated groups. Thus, although buparvaquone did not eliminate the parasites infecting the CNS, the drug represents an interesting lead with the potential to eliminate, or at least diminish, fetal infection during pregnancy. PMID:25941626

  9. In Vitro Activities of Combinations of Rifampin with Other Antimicrobials against Multidrug-Resistant Acinetobacter baumannii

    PubMed Central

    Bai, Yan; Liu, Bin; Wang, Tianlin; Cai, Yun; Liang, Beibei; Liu, Youning; Wang, Jin

    2014-01-01

    The antimicrobial treatment of multidrug-resistant (MDR) Acinetobacter baumannii infections has become a great challenge for medical staff all over the world. Increasing numbers of MDR A. baumannii infections have been identified and reported, but effective clinical treatments for them are decreasing. The objective of this study was to investigate the in vitro activities of combinations of rifampin (an established antimicrobial) and other antimicrobials, including biapenem, colistin, and tigecycline, against 73 clinical isolates of MDR A. baumannii. In total, 73 clinical isolates of MDR A. baumannii were collected from two A-level general hospitals in Beijing, and the MICs of rifampin, biapenem, colistin, and tigecycline were determined. The checkerboard method was used to determine the fractional inhibitory concentration indices (FICIs), that is, whether the combinations acted synergistically against these isolates. The MIC50, MIC90, and MICrange of rifampin combined with biapenem, colistin, and tigecycline against the isolates were clearly lower than those for four antimicrobials (rifampin, biapenem, colistin, and tigecycline) that were used alone. Combinations of rifampin with biapenem, colistin, and tigecycline individually demonstrated the following interactions: synergistic interactions (FICI ? 0.5) for 31.51%, 34.25%, and 31.51% of the isolates, partially synergistic interactions (0.5 < FICI < 1) for 49.31%, 43.83%, and 47.94% of the isolates, and additive interactions (FICI = 1) for 19.18%, 21.92%, and 20.55% of the isolates, respectively. There were no indifferent (1 < FICI < 4) or antagonistic (FICI ? 4) interactions. Therefore, combinations of rifampin with biapenem, colistin, or tigecycline may be future therapeutic alternatives for the treatment of MDR A. baumannii infections. PMID:25534730

  10. Antibiotic Activity against Naive and Induced Streptococcus pneumoniae Biofilms in an In Vitro Pharmacodynamic Model

    PubMed Central

    Vandevelde, Nathalie M.; Tulkens, Paul M.

    2014-01-01

    Biofilms play a role in the pathogenicity of pneumococcal infections. A pharmacodynamic in vitro model of biofilm was developed that allows characterization of the activity of antibiotics against viability and biomass by using in parallel capsulated (ATCC 49619) and noncapsulated (R6) reference strains. Naive biofilms were obtained by incubating fresh planktonic cultures for 2 to 11 days in 96-well polystyrene plates. Induced biofilms were obtained using planktonic bacteria collected from the supernatant of 6-day-old naive biofilms. Biomass production was more rapid and intense in the induced model, but the levels were similar for both strains. Full concentration responses fitting sigmoidal regressions allowed calculation of maximal efficacies and relative potencies of drugs. All antibiotics tested (amoxicillin, clarithromycin, solithromycin, levofloxacin, and moxifloxacin) were more effective against young naive biofilms than against old or induced biofilms, except macrolides/ketolides, which were as effective at reducing viability in 2-day-old naive biofilms and in 11-day-old induced biofilms of R6. Macrolides/ketolides, however, were less potent than fluoroquinolones against R6 (approximately 5- to 20-fold-higher concentrations needed to reduction viability of 20%). However, at concentrations obtainable in epithelial lining fluid, the viabilities of mature or induced biofilms were reduced 15 to 45% (amoxicillin), 17 to 44% (macrolides/ketolides), and 12 to 64% (fluoroquinolones), and biomasses were reduced 5 to 45% (amoxicillin), 5 to 60% (macrolides/ketolides), and 10 to 76% (fluoroquinolones), with solithromycin and moxifloxacin being the most effective and the most potent agents (due to lower MICs) in their respective classes. This study allowed the ranking of antibiotics with respect to their potential effectiveness in biofilm-related infections, underlining the need to search for still more effective options. PMID:24342635

  11. Glufosinate binds N-methyl-D-aspartate receptors and increases neuronal network activity in vitro.

    PubMed

    Lantz, Stephen R; Mack, Cina M; Wallace, Kathleen; Key, Ellen F; Shafer, Timothy J; Casida, John E

    2014-12-01

    Glufosinate (GLF) at high levels in mammals causes convulsions and amnesia through a mechanism that is not completely understood. The structural similarity of GLF to glutamate (GLU) implicates the glutamatergic system as a target for GLF neurotoxicity. The current work examined in vitro GLF interaction with N-methyl-D-aspartate subtype GLU receptors (NMDARs) and GLT-1 transporters via [(3)H]CGP 39653 binding experiments and [(3)H]GLU uptake assays, respectively. GLF effects on neuronal network activity were assessed using microelectrode array (MEA) recordings in primary cultures of cortical neurons. GLF and its primary metabolite N-acetylglufosinate (NAcGLF) bind to the NMDAR; the IC50 value for GLF was 668 ?M and for NAcGLF was about 100 ?M. Concentrations of GLF greater than 1000 ?M were needed to decrease GLU uptake through GLT-1. In MEA recordings from networks of rat primary cortical neurons, the concentration-responses for NMDA, GLF and NAcGLF on network mean firing rates (MFR) were biphasic, increasing at lower concentrations and decreasing below control levels at higher concentrations. Increases in MFR occurred between 3-10 ?M NMDA (290% control, maximum), 100-300 ?M NAcGLF (190% control, maximum) and 10-1000 ?M GLF (340% control, maximum). The NMDAR antagonist MK801 attenuated both NMDA and GLF increases in MFR. The GLF concentration required to alter GLU transport through GLT-1 is not likely to be attained in vivo, and therefore not relevant to the neurotoxic mode of action. However, toxicokinetic data from reports of intentional human poisonings indicate that GLF concentrations in the CNS after acute exposure could reach levels high enough to lead to effects mediated via NMDARs. Furthermore, the newly characterized action of NAcGLF at the NMDAR suggests that both the parent compound and metabolite could contribute to neurotoxicity via this pathway. PMID:25268653

  12. Comparison of in vitro activity of metronidazole and garlic-based product (Tomex®) on Trichomonas vaginalis.

    PubMed

    Ibrahim, Ayman Nabil

    2013-05-01

    Trichomonas vaginalis is a common sexually transmitted parasite in humans. Metronidazole has been the gold standard for treatment of trichomoniasis. The prevalence of metronidazole resistance and its unpleasant adverse effects drew the attention to the investigation of other lines of treatment, as that of herbal medicine. Garlic has been proven to have antibacterial, antiprotozoal, and antihelminthic activity. The current study was carried out to evaluate the efficacy of commercially available garlic (Tomex®) on T. vaginalis in vitro. The effect of different concentrations of garlic (12.5, 25, 50, and 100 ?g/ml) was determined on multiplication and motility of trophozoites at different time points (after 24, 48, 72, and 96 h) in comparison to the same concentrations of metronidazole at the same different time points. The results showed that parasite multiplication inhibition was noticed in proportion of concentration of Tomex and incubation time. The minimal lethal concentration of Tomex was 100 ?g/ml after 24 h, 50 ?g/ml after 48 h, 25 ?g/ml after 72 h, and 12.5 ?g/ml after 96 h. These results were similar to that of metronidazole as its minimal lethal concentration was 50 ?g/ml after 24 and 48 h and 12.5 ?g/ml after 72 and 96 h. Garlic had completely inhibited the motility of trophozoites with concentration of 100 ?g/ml after 24 h, 50 ?g/ml after 48 h, 25 ?g/ml after 72 h, and 12.5 ?g/ml after 96 h while metronidazole had completely inhibited the motility of trophozoites with concentration of 50 ?g/ml after 24 h, 25 ?g/ml after 48 h, and 12.5 ?g/ml after 72 and 96 h. This suggests that commercially available garlic (Tomex®) may be a promising phytotherapeutic agent for trichomoniasis. PMID:23455944

  13. Reliability analysis of two methods for measuring active enamel demineralization: An in vitro study

    PubMed Central

    Nassur, Camila; Pomarico, Luciana; Maia, Lucianne Cople

    2013-01-01

    Objective: The aim was to evaluate the intra- and inter-examiner reliability of two methods, namely, digital caliper [DC] and computerized image analysis software (Image Tool [IT] version 4.1) for measuring the area of an active enamel demineralization lesion. The effect on that measurement of a window that delimits the lesion was also assessed. Materials and Methods: Thirty-eight circular artificial enamel demineralization lesions (r = 2 mm) were produced on bovine teeth in vitro, which were then measured three times by three examiners, with and without the presence of a window delimiting the area to be measured. The data were analyzed by the intraclass correlation coefficient (ICC) with 95% confidence interval (CI) and by the Altman and Bland analysis. Results: The intra-examiner correlation using the IT method showed excellent reproducibility (mean ICC values 0.922-0.970 with the delimiting window and 0.915-0.990 without the window). However, the intra-examiner correlation using the DC method showed comparatively less reproducibility in the measurements (mean ICC values 0.458-0.648 with the delimiting window and 0.378-0.665 without the window). The inter-examiner correlations showed very good reproducibility of the measurements regardless of the presence or absence of the window for both the DC method (0.811-0.846) and IT method (0.953-0.994). However, the latter method showed less variability within the measurements. Conclusion: However, statistically no significant difference was found between both methodologies. Nevertheless, computerized image analysis with the Image Tool software demonstrated higher intra- and inter-examiner reliability than the digital caliper method, in estimating the area of the enamel demineralization lesion, regardless of the delimiting window. PMID:24883020

  14. Anti-hepatoma activity of a novel compound glaucocalyxin H in vivo and in vitro.

    PubMed

    Hai, Guangfan; Zhang, Chong; Jia, Yanlong; Bai, Suping; Han, Jinfen; Guo, Lanqing; Cui, Taizhen; Niu, Bingxuan; Huang, Feng; Song, Yu

    2015-06-01

    Glaucocalyxin H (GLH) is a new compound isolated from a traditional Chinese medical herb Isodon japonica var. glaucocalyx which has been used for folk medicine. This study was carried out for the first time to investigate the potential role of GLH in anti-hepatoma activity and underlying mechanisms in it. GLH could inhibit the growth of tumor in mice and induce HepG2 cells to death as assessed by the tumor reduction assay, toxic assay, morphological change, and survival rate assay. Many antitumor drugs originated from plants could inhibit the growth of tumor by inducing cells to apoptosis. The morphological changes of HepG2 cells treated with different concentrations of GLH under fluorescence and electron microscope and apoptotic rates were detected to verify its effect on apoptosis. As shown in the study, GLH could induce HepG2 cells to apoptosis in a dose-dependent manner. Bcl2 and Bax proteins played important roles in apoptosis and the disequilibrium between Bcl2 and Bax might result in apoptosis. The expression of Bax protein was upregulated and Bcl2 protein was downregulated in HepG2 cells treated with GLH assessed by Western blotting, and they were in a dose-dependent manner. Taken together, GLH can inhibit the growth of hepatoma cells in vivo and in vitro by inducing cell apoptosis due to the decreased Bcl2 and increased Bax proteins suggesting that GLH could be a potential candidate as an anti-hepatoma agent for the therapeutic treatment of hepatoma. PMID:25374342

  15. Nuclear activity of sperm cells during Hyacinthus orientalis L. in vitro pollen tube growth

    PubMed Central

    Zienkiewicz, Krzysztof; Suwi?ska, Anna; Niedojad?o, Katarzyna; Zienkiewicz, Agnieszka; Bednarska, El?bieta

    2011-01-01

    In this study, the transcriptional state and distribution of RNA polymerase II, pre-mRNA splicing machinery elements, and rRNA transcripts were investigated in the sperm cells of Hyacinthus orientalis L. during in vitro pollen tube growth. During the second pollen mitosis, no nascent transcripts were observed in the area of the dividing generative cell, whereas the splicing factors were present and their pools were divided between newly formed sperm cells. Just after their origin, the sperm cells were shown to synthesize new RNA, although at a markedly lower level than the vegetative nucleus. The occurrence of RNA synthesis was accompanied by the presence of RNA polymerase II and a rich pool of splicing machinery elements. Differences in the spatial pattern of pre-mRNA splicing factors localization reflect different levels of RNA synthesis in the vegetative nucleus and sperm nuclei. In the vegetative nucleus, they were localized homogenously, whereas in the sperm nuclei a mainly speckled pattern of small nuclear RNA with a trimethylguanosine cap (TMG snRNA) and SC35 protein distribution was observed. As pollen tube growth proceeded, inhibition of RNA synthesis in the sperm nuclei was observed, which was accompanied by a gradual elimination of the splicing factors. In addition, analysis of rRNA localization indicated that the sperm nuclei are likely to synthesize some pool of rRNA at the later steps of pollen tube. It is proposed that the described changes in the nuclear activity of H. orientalis sperm cells reflect their maturation process during pollen tube growth, and that mature sperm cells do not carry into the zygote the nascent transcripts or the splicing machinery elements. PMID:21081664

  16. Short-Term PTEN Inhibition Improves In Vitro Activation of Primordial Follicles, Preserves Follicular Viability, and Restores AMH Levels in Cryopreserved Ovarian Tissue From Cancer Patients

    PubMed Central

    Novella-Maestre, Edurne; Herraiz, Sonia; Rodríguez-Iglesias, Beatriz; Díaz-García, César; Pellicer, Antonio

    2015-01-01

    Introduction In vitro activation and growth of primordial dormant follicles to produce fertilizable oocytes would provide a useful instrument for fertility preservation. The employment of Phosphatase and TENsin homolog (PTEN) inhibitors, in combination with Protein kinase B (Akt) stimulating molecules, has been previously employed to increase follicular activation through the stimulation of the PTEN-Akt pathway. Methods We aim to establish improved in vitro activation also for cancer patients whose ovarian tissue has already been cryopreserved. Fresh and previously cryopreserved human ovarian cortex were exposed to short-term, low-concentration and ovary-specific treatment with only a PTEN inhibitor. Results Our in vitro activation protocol enhances the activation mechanisms of primordial follicles in both fresh and cryopreserved samples, and enlarges growing populations without inducing apoptosis in either follicles or the surrounding stroma. Treatment augments estradiol secretion and restores the expression levels of the previously diminished Anti-Müllerian hormone by means of cryopreservation procedures. Genomic modulation of the relative expression of PTEN pathway genes was found in treated samples. Conclusion The in vitro activation protocol offers new alternatives for patients with cryopreserved tissue as it increases the pool of viable activated follicles available for in vitro growth procedures. The combination of ovarian tissue cryopreservation and in vitro activation of primordial follicles, the main ovarian reserve component, will be a major advancement in fertility preservation. PMID:26024525

  17. In vitro chemopreventive activity of Camellia sinensis, Ilex paraguariensis and Ardisia compressa tea extracts and selected polyphenols.

    PubMed

    Ramirez-Mares, Marco Vinicio; Chandra, Sonia; de Mejia, Elvira Gonzalez

    2004-10-01

    Several herbal teas contain bioactive compounds that have been associated with a lower risk of chronic diseases including cancer. The aim of this study was to evaluate the chemopreventive activity of tea aqueous extracts and selected constituent pure polyphenols using a battery of in vitro marker systems relevant for the prevention of cancer. The effects of (-) epigallocatechin gallate (EGCG), quercetin (Q), gallic acid (GA), green tea (GT, Camellia sinensis), ardisia tea (AT, Ardisia compressa) and mate tea (MT, Ilex paraguariensis) extracts were tested. Cytotoxicity, TPA-induced ornithine decarboxylase (ODC) and quinone reductase (QR) activities were evaluated in vitro using HepG2 cells. The topoisomerase inhibitory activity was also tested, using the Saccharomyces cerevisiae yeast system. Results suggest that MT, AT and GT are cytotoxic to the HepG2 cells, with MT demonstrating dominant cytotoxicity. EGCG showed greater cytotoxicity than Q and GA against HepG2 cells. The greatest inhibition (82%) of TPA-induced ODC activity was shown by Q, with 25 microM (IC50 = 11.90 microM). Topoisomerase II, but not topoisomerase I, was the cellular target of MT, AT, EGCG, Q and GA, which acted mainly as true catalytic inhibitors. The cytotoxic activity and the inhibition of topoisomerase II may contribute to the overall chemopreventive activity of AT and MT extracts. Ardisia and mate teas may thus share a public health potential as chemopreventive agents. PMID:15450404

  18. Synthesis, DFT and antimicrobial activity assays in vitro for novel cis/trans-but-2-enedioic acid esters

    NASA Astrophysics Data System (ADS)

    Ma, Yan-Long; Zhou, Ru-Jin; Zeng, Xing-Ye; An, Ya-Xiong; Qiu, Song-Shan; Nie, Li-Jun

    2014-04-01

    Six novel cis/trans-but-2-enedioic acid esters had been synthesized to discover the new bioactive molecules that could kill food-related bacteria and fungi. Their structures were analyzed by melting point, LC-MS, 1H NMR and 13C NMR. 4-(Methoxycarbonyl) phenyl ethyl fumarate (6b) was also characterized by single-crystal X-ray diffraction. Their antimicrobial activities were evaluated in vitro by measuring the minimal inhibitory concentration (MIC). Compared with the single monomethyl fumarate and methyl 4-hydroxybenzoate, these compounds had stronger antimicrobial activity against all the eight microorganisms. Among the antibacterial and antifungal compounds, 4-(methoxycarbonyl) phenyl methyl fumarate (6a) showed the best antimicrobial activity. The electronic properties of these compounds were calculated by the density functional theory (DFT) method with 6-31G (d, p) basis set. DFT studies indicated that molecular electrostatic potential (MEP) map, ELUMO, energy gap, electronegativity and electrophilicity index could be helpful to understand the various antimicrobial activities among these compounds. The antimicrobial activity of compound 6a was evaluated in vitro against Salmonellacholeraesuis subsp. choleraesuis, Lactococcus lactis subsp. lactis and Saccharomyces cerevisiae by time-kill, and it was found that compound 6a exhibited significant microbiocidal activity against the three microorganisms.

  19. Activation by zinc of the human gastrin gene promoter in colon cancer cells in vitro and in vivo.

    PubMed

    Marshall, Kathryn M; Laval, Marie; Estacio, Ortis; Hudson, Damien F; Kalitsis, Paul; Shulkes, Arthur; Baldwin, Graham S; Patel, Oneel

    2015-10-01

    Over-expression of growth factors can contribute to the development and progression of cancer, and gastrins in particular have been implicated in accelerating the development of gastrointestinal cancers. Previously our group showed that hypoxia, cobalt chloride (a hypoxia mimetic) and zinc chloride could activate the expression of the gastrin gene in vitro. To characterise activation of the gastrin promoter by zinc ions further in vivo, TALEN technology was used to engineer a luciferase reporter construct into the endogenous human gastrin gene promoter in SW480 colon cancer cells. Gastrin promoter activity in the resultant Gast(luc) SW480 colon cancer cells was then measured by bioluminescence in cell culture and in tumour xenografts in SCID mice. Activation of intracellular signalling pathways was assessed by Western blotting. Activation of the gastrin promoter by zinc ions was concentration dependent in vitro and in vivo. Zinc ions significantly stimulated phosphorylation of ERK1/2 (MAPK pathway) but not of Akt (PI3K pathway). We conclude that the endogenous gastrin promoter is responsive to zinc ions, likely via activation of the MAPK pathway. PMID:26404630

  20. Blood shizonticidal activities of phenazines and naphthoquinoidal compounds against Plasmodium falciparum in vitro and in mice malaria studies

    PubMed Central

    de Souza, Nicolli Bellotti; de Andrade, Isabel M; Carneiro, Paula F; Jardim, Guilherme AM; de Melo, Isadora MM; da Silva, Eufrânio N; Krettli, Antoniana Ursine

    2014-01-01

    Due to the recent advances of atovaquone, a naphthoquinone, through clinical trials as treatment for malarial infection, 19 quinone derivatives with previously reported structures were also evaluated for blood schizonticide activity against the malaria parasite Plasmodium falciparum. These compounds include 2-hydroxy-3-methylamino naphthoquinones (2-9), lapachol (10), nor-lapachol (11), iso-lapachol (12), phthiocol (13) and phenazines (12-20). Their cytotoxicities were also evaluated against human hepatoma and normal monkey kidney cell lines. Compounds 2 and 5 showed the highest activity against P. falciparum chloroquine-resistant blood-stage parasites (clone W2), indicated by their low inhibitory concentration for 50% (IC50) of parasite growth. The therapeutic potential of the active compounds was evaluated according to the selectivity index, which is a ratio of the cytotoxicity minimum lethal dose which eliminates 50% of cells and the in vitro IC50. Naphthoquinones 2 and 5, with activities similar to the reference antimalarial chloroquine, were also active against malaria in mice and suppressed parasitaemia by more than 60% in contrast to compound 11 which was inactive. Based on their in vitro and in vivo activities, compounds 2 and 5 are considered promising molecules for antimalarial treatment and warrant further study. PMID:25099332

  1. Blood shizonticidal activities of phenazines and naphthoquinoidal compounds against Plasmodium falciparum in vitro and in mice malaria studies.

    PubMed

    de Souza, Nicolli Bellotti; de Andrade, Isabel M; Carneiro, Paula F; Jardim, Guilherme A M; de Melo, Isadora M M; da Silva Júnior, Eufrânio N; Krettli, Antoniana Ursine

    2014-08-01

    Due to the recent advances of atovaquone, a naphthoquinone, through clinical trials as treatment for malarial infection, 19 quinone derivatives with previously reported structures were also evaluated for blood schizonticide activity against the malaria parasite Plasmodium falciparum. These compounds include 2-hydroxy-3-methylamino naphthoquinones (2-9), lapachol (10), nor-lapachol (11), iso-lapachol (12), phthiocol (13) and phenazines (12-20). Their cytotoxicities were also evaluated against human hepatoma and normal monkey kidney cell lines. Compounds 2 and 5 showed the highest activity against P. falciparum chloroquine-resistant blood-stage parasites (clone W2), indicated by their low inhibitory concentration for 50% (IC50) of parasite growth. The therapeutic potential of the active compounds was evaluated according to the selectivity index, which is a ratio of the cytotoxicity minimum lethal dose which eliminates 50% of cells and the in vitro IC50. Naphthoquinones 2 and 5, with activities similar to the reference antimalarial chloroquine, were also active against malaria in mice and suppressed parasitaemia by more than 60% in contrast to compound 11 which was inactive. Based on their in vitro and in vivo activities, compounds 2 and 5 are considered promising molecules for antimalarial treatment and warrant further study. PMID:25099332

  2. Studies on the In Vitro Antiproliferative, Antimicrobial, Antioxidant, and Acetylcholinesterase Inhibition Activities Associated with Chrysanthemum coronarium Essential Oil

    PubMed Central

    Bardaweel, Sanaa K.; Hudaib, Mohammad M.; Tawaha, Khaled A.; Bashatwah, Rasha M.

    2015-01-01

    The essential oil of the Jordanian Chrysanthemum coronarium L. (garland) was isolated by hydrodistillation from dried flowerheads material. The oil was essayed for its in vitro scavenging activity using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. The results demonstrate that the oil exhibits moderate radical scavenging activity relative to the strong antioxidant ascorbic acid. In addition, cholinesterase inhibitory activity of C. coronarium essential oil was evaluated for the first time. Applying Ellman's colorimetric method, interesting cholinesterase inhibitory activity, which is not dose dependent, was evident for the oil. Furthermore, antimicrobial activities of the oil against both Gram-negative and Gram-positive bacteria were evaluated. While it fails to inhibit Gram-negative bacteria growth, the antibacterial effects demonstrated by the oil were more pronounced against the Gram-positive strains. Moreover, the examined oil was assessed for its in vitro antiproliferative properties where it demonstrated variable activities towards different human cancer cell lines, of which the colon cancer was the most sensitive to the oil treatment. PMID:26290675

  3. In Vitro Glycoengineering of IgG1 and Its Effect on Fc Receptor Binding and ADCC Activity

    PubMed Central

    Thomann, Marco; Schlothauer, Tilman; Dashivets, Tetyana; Malik, Sebastian; Avenal, Cecile; Bulau, Patrick; Rüger, Petra; Reusch, Dietmar

    2015-01-01

    The importance and effect of Fc glycosylation of monoclonal antibodies with regard to biological activity is widely discussed and has been investigated in numerous studies. Fc glycosylation of monoclonal antibodies from current production systems is subject to batch-to-batch variability. If there are glycosylation changes between different batches, these changes are observed not only for one but multiple glycan species. Therefore, studying the effect of distinct Fc glycan species such as galactosylated and sialylated structures is challenging due to the lack of well-defined differences in glycan patterns of samples used. In this study, the influence of IgG1 Fc galactosylation and sialylation on its effector functions has been investigated using five different samples which were produced from one single drug substance batch by in vitro glycoengineering. This sample set comprises preparations with minimal and maximal galactosylation and different levels of sialylation of fully galactosylated Fc glycans. Among others, Roche developed the glycosyltransferase enzyme sialyltransferase which was used for the in vitro glycoengineering activities at medium scale. A variety of analytical assays, including Surface Plasmon Resonance and recently developed Fc?R affinity chromatography, as well as an optimized cell-based ADCC assay were applied to investigate the effect of Fc galactosylation and sialylation on the in vitro Fc?RI, IIa, and IIIa receptor binding and ADCC activity of IgG1. The results of our studies do not show an impact, neither positive nor negative, of sialic acid- containing Fc glycans of IgG1 on ADCC activity, Fc?RI, and RIIIa receptors, but a slightly improved binding to Fc?RIIa. Furthermore, we demonstrate a galactosylation-induced positive impact on the binding activity of the IgG1 to Fc?RIIa and Fc?RIIIa receptors and ADCC activity. PMID:26266936

  4. In vitro activation of retinal cells: estimating location of stimulated cell by using a mathematical model

    NASA Astrophysics Data System (ADS)

    Ziv, Ofer R.; Rizzo, Joseph F., III; Jensen, Ralph J.

    2005-03-01

    Activation of neurons at different depths within the retina and at various eccentricities from the stimulating electrode will presumably influence the visual percepts created by a retinal prosthesis. With an electrical prosthesis, neurons will be activated in relation to the stimulating charge that impacts their cell membranes. The common model used to predict charge density is Coulomb's law, also known as the square law. We propose a modified model that can be used to predict neuronal depth that takes into account: (1) finite dimensions related to the position and size of the stimulating and return electrodes and (2) two-dimensional displacements of neurons with respect to the electrodes, two factors that are not considered in the square law model. We tested our model by using in vitro physiological threshold data that we had obtained previously for eight OFF-center brisk-transient rabbit retinal ganglion cells. For our most spatially dense threshold data (25 µm increments up to 100 µm from the cell body), our model estimated the depth of one RGC to be 76 ± 76 µm versus 87 ± 62 µm (median: SD) for the square law model, respectively. This difference was not statistically significant. For the seven other RGCs for which we had obtained threshold data up to 800 µm from the cell body, the estimate of the RGC depth (using data obtained along the X axis) was 96 ± 74 versus 20 ± 20 µm for the square law and our modified model, respectively. Although this difference was not statistically significant (Student t-test: p = 0.12), our model provided median values much closer to the estimated depth of these RGCs (Gt25 µm). This more realistic estimate of cell depth predicted by our model is not unexpected in this latter data set because of the more spatially distributed threshold data points that were evaluated. Our model has theoretical advantages over the traditional square law model under certain conditions, especially when considering neurons that are horizontally displaced from the stimulating electrode. Our model would have to be tested with a larger threshold data pool to permit more conclusive statements about the relative value of our model versus the traditional square law model under special circumstances. This work was supported by the Department of Veterans Affairs.

  5. Glucocorticoid activity detected by in vivo zebrafish assay and in vitro glucocorticoid receptor bioassay at environmental relevant concentrations.

    PubMed

    Chen, Qiyu; Jia, Ai; Snyder, Shane A; Gong, Zhiyuan; Lam, Siew Hong

    2016-02-01

    Glucocorticoids are pharmaceutical contaminants of emerging concern due to their incomplete removal during wastewater treatment, increased presence in aquatic environment and their biological potency. The zebrafish is a popular model for aquatic toxicology and environmental risk assessment. This study aimed to determine if glucocorticoids at environmental concentrations would perturb expression of selected glucocorticoid-responsive genes in zebrafish and to investigate their potentials as an in vivo zebrafish assay in complementing in vitro glucocorticoid receptor bioassay. The relative expression of eleven glucocorticoid-responsive genes in zebrafish larvae and liver of adult male zebrafish exposed to three representative glucocorticoids (dexamethasone, prednisolone and triamcinolone) was determined. The expression of pepck, baiap2 and pxr was up-regulated in zebrafish larvae and the expression of baiap2, pxr and mmp-2 was up-regulated in adult zebrafish exposed to glucocorticoids at concentrations equivalent to total glucocorticoids reported in environmental samples. The responsiveness of the specific genes were sufficiently robust in zebrafish larvae exposed to a complex environmental sample detected with in vitro glucocorticoid activity equivalent to 478 pM dexamethasone (DEX-EQ) and confirmed to contain low concentration (0.2 ng/L or less) of the targeted glucocorticoids, and possibly other glucocorticoid-active compounds. The findings provided in vivo relevance to the in vitro glucocorticoid activity and suggested that the environmental sample can perturb glucocorticoid-responsive genes in its original, or half the diluted, concentration as may be found in the environment. The study demonstrated the important complementary roles of in vivo zebrafish and in vitro bioassays coupled with analytical chemistry in monitoring environmental glucocorticoid contaminants. PMID:26461441

  6. Curcumin regulates peroxisome proliferator-activated receptor-? coactivator-1? expression by AMPK pathway in hepatic stellate cells in vitro.

    PubMed

    Zhai, Xuguang; Qiao, Haowen; Guan, Wei; Li, Ziqiang; Cheng, Yuanyuan; Jia, Xin; Zhou, Yajun

    2015-01-01

    Curcumin exerts an inhibitory effect on hepatic stellate cell (HSC) activation, a key step for liver fibrogenesis, and on liver fibrosis by up-regulation of peroxisome proliferator-activated receptor-? (PPAR?) expression. PPAR? plays a crucial role in suppression of HSC activation. Peroxisome proliferator-activated receptor-? coactivator-1? (PGC-1?) functions as a co-activator for PPAR?. Therefore, researches on the effect of curcumin on PGC-1? might contribute to understanding of the mechanisms underlying curcumin inhibition of HSC activation and liver fibrosis through PPAR?. The present study aimed to investigate the effect of curcumin on PGC-1? expression in HSCs in vitro and examine the underlying molecular mechanisms by western blot, reat-time PCR, and transfection. Our results showed that curcumin stimulation increased PGC-1? expression and the effects of curcumin on PGC-1? expression were correlated with the activation of adenosine monophosphate-activated protein kinase (AMPK). Curcumin increased superoxide dimutase-2 (SOD2) transcription and activity by AMPK/PGC-1? axis. Moreover, PGC-1? was demonstrated to inhibit ?1(I) collagen (a marker for liver fibrosis) transcription in cultured HSCs. These results demonstrated the promotion effect of curcumin on PGC-1? expression through AMPK pathway, which led to the increases in PPAR? activity and in SOD-2 transcription and activity. These data might suggest a possible new explanation for the inhibitory effect of curcumin on HSC activation and on liver fibrogenesis. PMID:25445048

  7. In vitro antioxidant activity and HPTLC determination of n-hexane extract of Emilia sonchifolia (L.)DC.

    PubMed Central

    Sophia, D.; Ragavendran, P.; Arulraj, C.; Gopalakrishnan, V. K.

    2011-01-01

    The free radical scavenging activities of n-hexane extract of the whole plant of Emilia sonchifolia was evaluated by employing various in vitro assay systems like DPPH radical scavenging activity, superoxide radical scavenging activity and hydrogen peroxide scavenging activity with IC50 values 180, 160 and 160 ?g/ml respectively. The results of the study indicate that the n-hexane extract of the whole plant of Emilia sonchifolia possess a significant scavenging effect with increasing concentrations probably due to its antioxidant potential. High performance thin layer chromatography (HPTLC) analysis in the n-hexane extract of Emilia sonchifolia showed the presence of terpenoids which probably may be responsible for the antioxidant activity. Thus, n-hexane extract of Emilia sonchifolia can be used potentially as a bioactive source of natural antioxidants due to the presence of terpenoids in it PMID:24826021

  8. Comparative in vitro activity of the new oral cephalosporin Bay v 3522 against aerobic and anaerobic bacteria.

    PubMed

    Rylander, M; Nord, C E; Norrby, S R

    1990-10-01

    The in vitro activity of the new oral cephalosporin Bay v 3522 against 229 aerobic and 330 anaerobic clinical isolates was determined using the agar dilution technique. For comparison, amoxicillin, amoxicillin/clavulanate, cefaclor, cefadroxil, cefuroxime, cephalexin, ciprofloxacin, clindamycin, co-trimoxazole, doxycycline, erythromycin and metronidazole (only anaerobic bacteria) were tested. Bay v 3522 was found to have high activity against Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Streptococcus pneumoniae, Streptococcus pyogenes, Branhamella catarrhalis, Haemophilus influenzae, anaerobic cocci, Propionibacterium acnes, Clostridium perfringens and fusobacteria. When tested against a higher inoculum or using the broth dilution technique, the activity of Bay v 3522 showed little dependence on inoculum size and the bactericidal activity was similar to inhibitory activity in most bacterial groups. Bay v 3522 may be useful in the treatment of skin, soft tissue and respiratory tract infections. Clinical studies are thus warranted. PMID:2261923

  9. Assessment of Anti-nutritive Activity of Tannins in Tea By-products Based on In vitro Rumen Fermentation

    PubMed Central

    Kondo, Makoto; Hirano, Yoshiaki; Ikai, Noriyuki; Kita, Kazumi; Jayanegara, Anuraga; Yokota, Hiro-omi

    2014-01-01

    Nutritive values of green and black tea by-products and anti-nutritive activity of their tannins were evaluated in an in vitro rumen fermentation using various molecular weights of polyethylene glycols (PEG), polyvinyl pyrrolidone (PVP) and polyvinyl polypyrrolidone as tannin-binding agents. Significant improvement in gas production by addition of PEG4000, 6000 and 20000 and PVP was observed only from black tea by-product, but not from green tea by-product. All tannin binding agents increased NH3-N concentration from both green and black tea by-products in the fermentation medium, and the PEG6000 and 20000 showed relatively higher improvement in the NH3-N concentration. The PEG6000 and 20000 also improved in vitro organic matter digestibility and metabolizable energy contents of both tea by-products. It was concluded that high molecular PEG would be suitable to assess the suppressive activity of tannins in tea by-products by in vitro fermentation. Higher responses to gas production and NH3-N concentration from black tea by-product than green tea by-product due to PEG indicate that tannins in black tea by-product could suppress rumen fermentation more strongly than that in green tea by-product. PMID:25358316

  10. In vitro studies of acute toxicity mechanisms and structure-activity relationships of nonionic surfactants in fish

    SciTech Connect

    Bodishbauah, D.F.

    1994-12-31

    In fish, gills are believed to be a primary target for a number of toxicants. Gills perform the essential systemic functions of gas exchange, waste elimination, and ion/pH balance, and are exposed to ambient environmental toxicant levels. Qualitative gill morphology changes are easily observed, but quantitative measures of impaired function are difficult. This in vitro technique utilizes the opercular epithelium of the mummichog, Fundulus heteroclitus, as a surrogate for gill epithelium in mechanistic toxicity and structure-activity studies. This model has long been used by electrophysiologists studying osmoregulation in marine fish. Effects on trans-epithelial potential (TEP) and/or short-circuit current (I{sub sc}) across the opercular epithelium can be made for any pollutant of interest, using an epithelial voltage clamp and Ussing chamber. The nonionic synthetic surfactant class, alkylphenol ethoxylates, were chosen as a model toxicant class to test this experimental model. Synthetic surfactants are ubiquitous waterborne pollutants, with annual North American usage approaching eight billion pounds. Surfactants are recognized as potent, acute gill toxicants in fish. The exact mechanism of toxicity has yet to be elucidated. These compounds proved to be potent inhibitors of both TEP and I{sub sc} in vitro, at dose levels comparable to those causing lethality, suggesting that impaired osmoregulation plays a role in their acute toxicity. Similar structure-activity relationships were found for the endpoints of acute lethality to F. heteroclitus and impaired in vitro epithelial transport.

  11. Prebiotic Potential of Xylooligosaccharides Derived from Corn Cobs and Their In Vitro Antioxidant Activity When Combined with Lactobacillus.

    PubMed

    Yu, Xiuhua; Yin, Jianyuan; Li, Lin; Luan, Chang; Zhang, Jian; Zhao, Chunfang; Li, Shengyu

    2015-07-01

    In the present work, the in vitro prebiotic activity of xylooligosaccharides (XOS) derived from corn cobs combined with Lactobacillus plantarum, a probiotic microorganism, was determined. These probiotics exhibited different growth characteristics depending on strain specificity. L. plantarum S2 cells were denser and their growth rates were higher when cultured on XOS. Acetate was found to be the major short-chain fatty acid produced as the end-product of fermentation, and its amount varied from 1.50 to 1.78 mg/ml. The antimicrobial activity of XOS combined with L. plantarum S2 was determined against gastrointestinal pathogens. The results showed that XOS proved to be an effective substrate, enhancing antimicrobial activity for L. plantarum S2. In vivo evaluation of the influence of XOS and L. plantarum S2, used both alone and together, on the intestinal microbiota in a mouse model showed that XOS combined with L. plantarum S2 could increase the viable lactobacilli and bifidobacteria in mice feces and decrease the viable Enterococcus, Enterobacter, and Clostridia spp. Furthermore, in the in vitro antioxidant assay, XOS combined with L. plantarum S2 possessed significant 2,2-diphenyl-1- picrylhydrazyl, 2,2'-azino-bis, and superoxide anion radical-scavenging activities, and the combinations showed better antioxidant activity than either XOS or L. plantarum S2 alone. PMID:25791856

  12. Antitumor Activities of Ethyl Acetate Extracts from Selaginella doederleinii Hieron In Vitro and In Vivo and Its Possible Mechanism

    PubMed Central

    Li, Juan; Zhao, Ping; Ma, Wen-tao; Feng, Xie-he; Chen, Ke-li

    2015-01-01

    The antitumor activities of ethyl acetate extracts from Selaginella doederleinii Hieron (SD extracts) in vitro and in vivo and its possible mechanism were investigated. HPLC method was developed for chemical analysis. SD extracts were submitted to 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay on different cells, flow cytometry, and RT-PCR analysis using HepG2 cell and antitumor activity in vivo using H-22 xenograft tumor mice. Six biflavonoids from SD extracts were submitted to molecular docking assay. The results showed that SD extracts had considerable antitumor activity in vitro and in vivo without obvious toxicity on normal cells and could induce cell apoptosis. The mechanisms of tumorigenesis and cell apoptosis induced by SD extracts may be associated with decreasing the ratio of bcl-2 and bax mRNA level, activating caspase-3, suppressing survivin, and decreasing the gene expression of COX-2, 5-LOX, FLAP, and 12-LOX mRNA. The main active component in SD extracts is biflavonoids and some exhibited strong interactions with COX-2, 5-LOX, 12-LOX, and 15-LOX. These results offering evidence of possible mechanisms of SD extracts suppress cell proliferation and promote apoptosis and provide the molecular theoretical basis of clinical application of S. doederleinii for cancer therapy. PMID:25866543

  13. Temperature-dependent in vitro antimicrobial activity of four 4-quinolones and oxytetracycline against bacteria pathogenic to fish.

    PubMed Central

    Martinsen, B; Oppegaard, H; Wichstrřm, R; Myhr, E

    1992-01-01

    The in vitro antimicrobial activities of oxolinic acid, flumequine, sarafloxacin, enrofloxacin, and oxytetracycline against strains of bacteria pathogenic to fish (Aeromonas salmonicida subsp. salmonicida, atypical A. salmonicida, Vibrio salmonicida, Vibrio anguillarum, and Yersinia ruckeri) were determined at two different incubation temperatures, 4 and 15 degrees C, by a drug microdilution method. The main objective of the study was to examine the effect of incubation temperature on the in vitro activities of 4-quinolones and oxytetracycline against these bacteria. When tested against A. salmonicida subsp. salmonicida, all of the quinolones examined had MICs two- to threefold higher at 4 degrees C than at 15 degrees C. Similarly, 1.5- to 2-fold higher MICs were recorded for all of the quinolones except sarafloxacin at 4 degrees C than at 15 degrees C when the drugs were tested against V. salmonicida. In contrast to those of the quinolones, the MICs of oxytetracycline were two- to eightfold lower at 4 degrees C than at 15 degrees C against all of the bacterial species tested. Of the antimicrobial agents tested against the bacterial species included in the study, enrofloxacin was the most active and oxytetracycline was the least active. Sarafloxacin was slightly more active than flumequine and oxolinic acid, especially against oxolinic acid-resistant A. salmonicida subsp. salmonicida strains. PMID:1416857

  14. International Regulations of Propolis Quality: Required Assays do not Necessarily Reflect their Polyphenolic-Related In Vitro Activities.

    PubMed

    Bridi, Raquel; Montenegro, Gloria; Nuńez-Quijada, Gabriel; Giordano, Ady; Fernanda Morán-Romero, Maria; Jara-Pezoa, Isaac; Speisky, Hernán; Atala, Elias; López-Alarcón, Camilo

    2015-06-01

    Propolis has been proposed as a polyphenolic-rich natural product potentially able to be used for human consumption or even for medicinal proposes. To guarantee a minimum phenolic and flavonoid content and as consequence of their related-biological activities, international requirements of propolis quality are commonly applied. In this work we assessed phenolic and flavonoid contents of propolis; the antioxidant capacity (toward peroxyl radicals and hypochlorous acid); the ability to generate nitric oxide (NO); and, finally the antimicrobial activity of 6 propolis samples from the VI region of Chile. Our results show that the total phenolic and flavonoid content of propolis samples are not always in agreement with their polyphenolic-associated in vitro activities. For example, P03 and P06 samples showed the lowest (25 ± 4 GAE/g propolis) and the highest (105 ± 3 GAE/g propolis) total phenolic content, respectively. This was in agreement with flavonoid content and their Oxygen Radical Absorbance Capacity (ORAC) activity. However, this dependence was not observed toward HOCl, NO release and antimicrobial activity. Based on our results, we consider that, in order to guarantee the antioxidant or antimicrobial in vitro effects, the international regulations of propolis quality should contemplate the convenience of incorporating other simple analytical test such as ORAC or antimicrobial tests. PMID:25944094