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H Incubation (h + incubation)

Distribution by Scientific Domains
Life Sciences40%
Medical Sciences23%
Chemistry21%
Earth and Environmental Science9%
Polymers and Materials Science5%
Distribution within Life Sciences
Biotechnology (Life Sciences)12%

Terms modified by H Incubation

  • h incubation period
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    Selected Abstracts

    Physiological and biochemical analyses of microcystin-RR toxicity to the cyanobacterium Synechococcus elongatus

    ENVIRONMENTAL TOXICOLOGY, Issue 6 2004
    Zhi-quan Hu
    Abstract Freshwater Microcystis may form dense blooms in eutrophic lakes. It is known to produce a family of related cyclic hepatopeptides (microcystins, MC) that constitute a threat to aquatic ecosystems. Most toxicological studies of microcystins have focused on aquatic animals and plants, with few examining the possible effects of microcystins on phytoplankton. In this study we chose the unicellular Synechococcus elongatus (one of the most studied and geographically most widely distributed cyanobacteria in the picoplankton) as the test material and investigated the biological parameters: growth, pigment (chlorophyll-a, phycocyanin), photosynthetic activity, nitrate reductase activity, and protein and carbohydrate content. The results revealed that microcystin-RR concentrations above 100 ,g · L,1 significantly inhibited the growth of Synechococcus elongatus. In addition, a change in color of the toxin-treated algae (chlorosis) was observed in the experiments. Furthermore, MC-RR markedly inhibited the synthesis of the pigments chlorophyll-a and phycocyanin. A drastic reduction in photochemical efficiency of PSII (Fv/Fm) was found after a 96-h incubation. Changes in protein and carbohydrate concentrations and in nitrate reductase activity also were observed during the exposure period. This study aimed to evaluate the mechanisms of microcystin toxicity on a cyanobacterium, according to the physiological and biochemical responses of Synechococcus elongatus to different doses of microcystin-RR. The ecological role of microcystins as an allelopathic substance also is discussed in the article. © 2004 Wiley Periodicals, Inc. Environ Toxicol 19: 571,577, 2004. [source]

    Apoptotic effect of cyanobacterial extract on rat hepatocytes and human lymphocytes

    ENVIRONMENTAL TOXICOLOGY, Issue 3 2001
    Joanna Mankiewicz
    Abstract Toxic cyanobacterial blooms are an increasing problem in Poland. The production of cyanobacterial toxins and their presence in drinking and recreational waters represent a growing danger to human and animal health. This is connected with the increase of cyanobacterial biomass caused by excessive eutrophication of the water ecosystem. There is evidence that cyanobacterial hepatotoxins can act as a potent promoter of primary liver cancer. The apoptotic effect of microcystins in Polish cyanobacterial bloom samples on rat hepatocytes and human lymphocytes was observed using light and fluorescence microscopy, flow cytometry, and electrophoretic analysis. The incubation time needed to observe the first morphological apoptotic changes in hepatocytes was approximately 30 min; however, the characteristic biochemical changes in DNA were not observed even after 120 min. In lymphocyte cultures the morphological changes characteristic for apoptosis were observed after 24 h of incubation and a 48-h incubation was found to be optimal for analysis of internucleosomal DNA fragmentation, which is one of the main biochemical hallmarks of programmed cell death. These cells are an easily isolated and inexpensive material for medical diagnostics. Therefore the apoptotic changes, together with the clastogenic effect seen in lymphocyte cultures, are proposed as a future analytical method for these toxins. © 2001 John Wiley & Sons, Inc. Environ Toxicol 16: 225,233, 2001 [source]

    Ecdysteroid synthesis and imaginal disc development in the midge Chironomus riparius as biomarkers for endocrine effects of tributyltin

    ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 5 2002
    Torsten Hahn
    Abstract Acute effects of the endocrine disruptor bis (tri- n -butyltin) oxide (TBTO) on molting-hormone biosynthesis and imaginaldisc development were investigated in larvae of the midge Chironomus riparius (Meigen). Ecdysteroid synthesis was measured by 24-h incubation of molting-hormone-synthesizing tissues (prothoracic glands) in vitro with or without the addition of TBTO. The amount of ecdysteroids produced was analyzed by radioimmunoassay. Developmental effects in vivo were investigated by determining the developmental phase of the genital imaginal discs before and after a 48-h exposure to TBTO in water. Sex-specific effects were found with both endpoints. Ecdysteroid synthesis was significantly reduced (analysis of variance [ANOVA], p , 0.005) in female larvae at all concentrations (TBTO-Sn at 50, 500, and 5,000 ng/L), whereas a significant elevation of the biosynthesis rate occurred in male larvae in the 500-ng/L treatment (ANOVA, p , 0.05). In vivo experiments with development of the genital imaginal disc within a 48-h exposure period revealed a significantly slower development in female larvae and a significantly faster development in male larvae (contingency tables, p , 0.001) at all concentrations tested (TBTO-Sn at 10, 50, 200, and 1,000 ng/L). These results partly coincided with the in vitro effects on molting-hormone synthesis. The 48-h median lethal concentration (LC50) was 25 ,g/L (20,30 ,g/L 95% confidence intervals). The combination of in vitro and in vivo methods has proven to be a useful approach for the detection of endocrine effects of TBTO in C. riparius at levels 2,000-fold below the LC50 value. High sensitivity and short test duration suggest that chironomids may have potential as freshwater sentinel organisms for endocrine-disrupting chemicals. [source]

    Microglial glutamate uptake is coupled to glutathione synthesis and glutamate release

    EUROPEAN JOURNAL OF NEUROSCIENCE, Issue 4 2006
    Mikael Persson
    Abstract The physiological function of microglial glutamate uptake has been debated as it is about 10% of that measured for astrocytes. This study addresses how glutamate, taken up from the extracellular space, is utilized by microglia. It was found that purified rat microglia incubated for 60 min with 3H-glutamate had an increased intracellular accumulation of 3H-glutamate after 12 h incubation with tumour necrosis factor alpha (TNF-,) but not after incubation with lipopolysaccharide (LPS). Furthermore, LPS- but not TNF-,-treated cells showed an increased efflux of 3H-labelled compounds, presumably glutamate through the XC, system and treatment with LPS or TNF-, increased the microglial glutathione concentrations and led to an increased incorporation of 3H-glutamate into glutathione. Depending on the stimuli, 3,6% of the total labelled contents were found in the form of glutathione and 25,35% in the form of glutamate. These results show that microglial glutamate uptake is directly coupled to glutathione synthesis and release of glutamate and/or glutamate metabolites. Additionally, the increased glutathione contents after LPS or TNF-, treatment were able to reduce microglial cell death after H2O2 challenge, showing a potential (self)-protective function for microglial glutamate transporter expression and glutathione synthesis. [source]

    Effects of Endotoxin Exposure on Cationic Amino Acid Transporter Function in Ovine Peripheral Blood Mononuclear Cells

    EXPERIMENTAL PHYSIOLOGY, Issue 2 2003
    Megan F. Clark
    Rodent models of sepsis differ from clinical human disease in that humans make substantially less whole-body nitric oxide and have different cellular responses to endotoxin. Sheep, when exposed to endotoxin, behave in a manner more similar to humans. Many studies of rodent peripheral blood mononuclear cells (PBMCs) exposed to endotoxin demonstrate increased cationic amino acid transporter function (particularly through the y+ transporter) to supply arginine substrate to upregulated nitric oxide synthase. Whether this is true in sheep is not known. We have studied cationic amino acid transport in sheep PBMCs stimulated with endotoxin, using labelled lysine. PBMCs stimulated both in vitro and in vivo show an initial reduction in total and y+ lysine transport (after 1-2 h exposure to endotoxin): a previously undescribed effect of endotoxin. In in vitro activated cells, the reduction in y+ transport was prevented by the lipoxygenase inhibitor, nordihydroguaretic acid (NDGA), and the phospholipase inhibitor 4-bromophenacyl bromide (4-BPAB), but not cyclohexamide or a number of other inhibitors of intracellular second-messenger pathways. In contrast after 14 h incubation, the expected increase in total and y+ lysine transport was seen. The increase in y+ transport could be prevented by cyclohexamide, dexamethasone, ibuprofen, the protein kinase C inhibitor sphingosine, NDGA and 4-BPAB. These results suggest that in response to endotoxin exposure there is an initial decrease in y+ activity mediated by a lipoxygenase product, followed by a substantial increase in y+ activity mediated by the products of either cyclo-oxygenase or lipoxygenase. Cyclo-oxygenase and/or lipoxygenase inhibition might be useful in reducing arginine transport, and hence nitric oxide production, in these cells. [source]

    Survival and vitality of Gremmeniella abietina on Pinus sylvestris slash in northern Sweden

    FOREST PATHOLOGY, Issue 6 2006
    J. Witzell
    Summary Survival and vitality of Gremmeniella abietina on Pinus sylvestris slash was studied in northern Sweden during 2003 and 2004. Once a month between September 2003 and April 2004, two to three trees were cut down and debranched. Shoots with pycnidia were sampled at the felling date and then at every consecutive month. The percentage of germinated conidia from each shoot was calculated after 24, 48 and 72 h incubation. The vitality of G. abietina pycnidia in the slash remained high the whole period. Intact pycnidia were found on slash several months after the time of conidial sporulation, which indicates that new pycnidia may be produced on dead pine branches. Sampling of shoots from slash on 13- to 18-month-old clear-cuts showed conidial germination capacity as high as in pycnidia collected in fresh slash. Due to survival of G. abietina in slash it is recommended to postpone planting of P. sylvestris seedlings in northern boreal areas to the third vegetation period after sanitary clear-cuts. Résumé La survie et la vitalité de Gremmeniella abietina dans des rémanents de Pinus sylvestris ont étéétudiées dans le nord de la Suède pendant les années 2003 et 2004. Une fois par mois entre septembre 2003 et avril 2004, 2 ou 3 arbres ont été abattus et ébranchés. Des pousses avec pycnides ont étééchantillonnées à la date d'abattage et les mois suivants. Le pourcentage de conidies germées a été calculé pour chaque pousse après 24, 48 et 72 heures d'incubation. La vitalité des pycnides de G. abietina dans les rémanents est restée élevée tout au long de la période. Des pycnides intactes ont été trouvées dans les rémanents plusieurs mois après la période de sporulation conidienne, ce qui suggère que de nouvelles pycnides peuvent être produites sur des branches mortes de pin. Des échantillonnages de pousses dans des rémanents de coupes rases réalisées 13,18 mois plus tôt ont montré une capacité de germination des conidies aussi élevée que dans les pycnides collectées dans des rémanents fraîchement coupés. Du fait de la survie de G. abietina dans les rémanents, il est conseillé de reporter la plantation des semis de P. sylvestris dans les zones septentrionales boréales à la troisième saison de végétation après les coupes sanitaires. Zusammenfassung Das Überleben und die Vitalität von Gremmeniella abietina auf Schlagabraum von Pinus sylvestris wurde in den Jahren 2003 und 2004 untersucht. Zwischen September 2003 und April 2004 wurden in jedem Monat einmal 2,3 Bäume gefällt und entastet. Zum Zeitpunkt des Fällens und in jedem folgenden Monat wurden Triebe mit Pyknidien gesammelt. Von jedem Trieb wurde die Keimrate der Konidien nach 24, 48 und 72 Stunden Inkubation bestimmt. Während der gesamten Beobachtungsdauer blieb die Vitalität der Pyknidien im Schlagabraum hoch. Mehrere Monate nach der Sporulation wurden intakte Pyknidien gefunden, ein Hinweis darauf, dass möglicherweise neue Pyknidien auf den toten Kiefernzweigen gebildet wurden. Auf dem Schlagabraum von 13,18 Monate alten Kahlschlägen war die Keimfähigkeit der Konidien ähnlich hoch wie bei Pyknidien von frischem Schlagabraum. Aufgrund des langen Überlebens von G. abietina in Schlagabraum wird für die nördlichen borealen Gebiete empfohlen, nach phytosanitären Kahlschlägen P. sylvestris -Sämlinge erst in der dritten Vegetationsperiode zu pflanzen. [source]

    Biotransformation in vitro of the 22R and 22S epimers of budesonide by human liver, bronchus, colonic mucosa and skin

    FUNDAMENTAL & CLINICAL PHARMACOLOGY, Issue 1 2001
    Julio Cortijo
    The pharmacological effects of glucocorticoids are greatly influenced by their pharmacokinetic properties. In the present report, the in vitro biotransformation of the 22R and 22S epimers of the topical steroid budesonide was studied in the S-9 fraction of human liver, bronchus, skin and colonic mucosa. The disappearance of unchanged epimers of budesonide was measured during 90 min of incubation by high performance liquid chromatography. The rate of disappearance was high in human liver while little biotransformation occurred in bronchial tissue and colonic mucosa, and none was detected in the skin. A marked decay of the initial concentration of unchanged budesonide epimers was noticed after 2 h incubation in cultured human hepatocytes, while only a small decrease was observed after 24 h incubation in cultured human airway smooth muscle cells and BEAS-2B cells. The 22R epimer of budesonide suffered greater in vitro biotransformation than the 22S epimer in human hepatic, bronchial and colonic tissues. These findings extend those of other studies, and confirm that the high therapeutic ratio of budesonide is due to negligible local biotransformation combined with high level of liver metabolism for locally absorbed budesonide. [source]

    In situ rumen degradation and in vitro gas production of some selected grains from Turkey

    JOURNAL OF ANIMAL PHYSIOLOGY AND NUTRITION, Issue 9-10 2002
    H. D. Umucalilar
    Summary An investigation of the dry matter degradability (DMD) and effective dry matter degradability (EDDM) was performed for barley, wheat, rye, corn, triticale and oat samples, using the Nylon-bag technique. Gas production (GP), metabolizable energy (ME) and in vitro organic matter digestibility (IVOMD) were also studied by using Hohenheim gas test. The DM from barley, wheat, rye and triticale was digested rapidly in the rumen, and, at the 48 h of incubation, degradability was found to be approximately about 80%. The higher degradability observed for these grains than for oats and corn was attributable to the structure of these grains. In contrast, DM of corn and oats was degraded very slowly and reached 66.7 and 66.5 at 48 h, respectively. Effective degradability values of barley, wheat, rye, corn, triticale and oats were determined to be 61.4, 69.0, 64.0, 41.7, 66.7 and 58.6% in 5% rumen outflow rate, respectively. At the end of the 48 h incubation, total gas productions in barley, wheat, rye, corn, triticale and oats were estimated to be 83.6, 87.2, 87.5, 83.5, 85.8 and 63.9 ml/200 mg DM, respectively. The mean ME values of these grains calculated from cumulative gas amount at 24 h incubation were 11.8, 12.1, 12.3, 10.9, 12.4 and 10.2 MJ/kg DM, respectively. In vitro digestible organic matter of barley, wheat, rye, corn, triticale and oats were estimated to be 85.0, 87.3, 88.2, 79.5, 89.0 and 72.6%. Percentage overall EDDM (k=5%) of barley, wheat, rye, triticale and oats was positively correlated with in vitro GP at 6 h, cumulative GP at 24 h and total GP at 48 h (p<0.05). As a result, in situ dry matter degradation of grains showed great differences depending on the chemical compositions. In situ EDDM of grains may be predicted from in vitro gas production parameters. [source]

    Prostaglandins in rainbow trout (Oncorhynchus mykiss Walbaum, 1792) sperm biology , searching for answers

    JOURNAL OF APPLIED ICHTHYOLOGY, Issue 4 2008
    R. K. Kowalski
    Summary The purpose of this study was to determine the concentrations of prostaglandins E2 and F2, (PGE2 and PGF2,) in the blood, testis and seminal plasma of mature male rainbow trout and in the ovarian fluid to assess the effects of these prostaglandins on sperm motility parameters when present in activation media. Also prolonged incubation with prostaglandins on sperm motility and calcium influx were studied. The profile of PGE2 and PGF2, differed in concentration between blood, testicular supernatant and seminal plasma. PGE2 was predominant in the blood sample (0.29 ng ml,1) and testicular supernatant (3.1 ng ml,1) whereas their level in seminal plasma was lower than PGF2, (0.23 ng ml,1). The concentrations of PGF2, in blood, testis and seminal plasma were 0.04, 0.99, 1.3 ng ml,1, respectively. In the ovarian fluid the concentrations of both prostaglandins were higher than in the male reproductive tract. Adding both prostaglandins to activation buffer (at concentrations 15 and 70 ng ml,1) had no effect on any CASA parameters. Calcium influx related to rainbow trout sperm incubations with PGE2, and PGF2, was not detected. After 24 h incubation of sperm in artificial seminal plasma solution without and with prostaglandins all sperm samples increased their motility potential and intracellular calcium concentration. Therefore, this effect was not related to the presence of prostaglandins. In summary PGE2, and PGF2, were present in the rainbow trout male reproductive tract, and their profile varies from that of blood, testis and seminal plasma. The specific role of both prostaglandins in salmonid sperm biology remains unclear. [source]

    Evaluation of novel fluorogenic substrates for the detection of glycosidases in Escherichia coli and enterococci

    JOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2006
    J.D. Perry
    Abstract Aims:, Enzyme substrates based on 4-methylumbelliferone are widely used for the detection of Escherichia coli and enterococci in water, by detection of , -glucuronidase and , -glucosidase activity respectively. This study aimed to synthesize and evaluate novel umbelliferone-based substrates with improved sensitivity for these two enzymes. Methods and Results:, A novel , -glucuronide derivative based on 6-chloro-4-methylumbelliferone (CMUG) was synthesized and compared with 4-methylumbelliferyl- , - d -glucuronide (MUG) using 42 strains of E. coli in a modified membrane lauryl sulfate broth. Over 7 h of incubation, the fluorescence generated from the hydrolysis of CMUG by E. coli was over twice that from MUG, and all of the 38 glucuronidase-positive strains generated a higher fluorescence with CMUG compared with MUG. Neither substrate caused inhibition of bacterial growth in any of the tested strains. Four , -glucosidase substrates were also synthesized and evaluated in comparison with 4-methylumbelliferyl- , - d -glucoside (MU-GLU) using 42 strains of enterococci in glucose azide broth. The four substrates comprised , -glucoside derivatives of umbelliferone-3-carboxylic acid and its methyl, ethyl and benzyl esters. Glucosides of the methyl, ethyl and benzyl esters of umbelliferone-3-carboxylic acid, were found to be superior to MU-GLU for the detection of enterococci, especially after 18 h of incubation, while umbelliferone-3-carboxylic acid- , - d -glucoside was inferior. However, the variability in detectable , -glucosidase activity among the different strains of enterococci in short-term assays using the three carboxylate esters (7 h incubation) may compromise their use for rapid detection and enumeration of these faecal indicator bacteria. Conclusions:, The , -glucuronidase substrate CMUG appears to be a more promising detection system than the various , -glucosidase substrates tested. Significance and Impact of the Study:, The novel substrate CMUG showed enhanced sensitivity for the detection of , -glucuronidase-producing bacteria such as E. coli, with a clear potential for application in rapid assays for the detection of this indicator organism in natural water and other environmental samples. [source]

    Comparison of the sensitivity of manual and automated immunomagnetic separation methods for detection of Shiga toxin-producing Escherichia coli O157:H7 in milk

    JOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2002
    R.D. Reinders
    Aim:,To determine the sensitivity of methods for detection of injured and uninjured Escherichia coli O157:H7 (E. coli O157) in raw and pasteurized milk. Methods and Results:,Raw milk, pasteurized milk with 1·5% fat content and pasteurized milk with 3·5% fat content were spiked with E. coli O157 at low levels. The samples were enriched in modified tryptone soya broth with novobiocin (mTSBn) at 37°C. Aliquots of the enriched culture were analysed either by manual immunomagnetic separation (MIMS) and culturing on sorbitol MacConkey agar with or without cefixime and potassium tellurite (SMACct or SMAC), or by automated immunomagnetic separation and integrated ELISA (EiaFossÔ). Uninjured E. coli O157 organisms were detected in milk by both methods at 1 cfu 10 ml,1 sample). Injured organisms were detected at levels of about 4 cfu 10 ml,1 sample. Direct enrichment in mTSBn (22 h incubation) showed better sensitivity for injured cells than enrichment in buffered peptone water (BPW, 22 h incubation), or in a two-step enrichment consisting of BPW (6 h, 37°C) and mTSBn (16 h, 37°C), successively. Conclusions:,The methods showed equal sensitivity in that they were both able to detect 1 cfu 10 ml,1 milk sample. Injured organisms can be detected and isolated at a level almost as low as this. A resuscitation step is not recommended for the detection and isolation of injured and non-injured E. coli O157 from milk. Significance and Impact of the Study:,Due to the dilution of contamination in the bulk tank, analysis of milk for the presence of E. coli O157 requires a very sensitive method. Both methods described here are useful for such analysis. [source]

    Comparison of ATP and in vivo bioluminescence for assessing the efficiency of immunomagnetic sorbents for live Escherichia coli O157:H7 cells

    JOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2002
    W. Sun
    Aims:,To develop methods to assess the efficiency of immunomagnetic separation (IMS). Methods and Results:,The capturing efficiency of biosorbents for Escherichia coli O157:H7, constructed using streptavidin-coated magnetic beads and biotinylated antibodies, was tested using both in vivo and ATP bioluminescence. Both methods were suitable for the enumeration of bacteria captured by the biosorbents. The level of both ATP and in vivo bioluminescence depended on the media used, but was unaffected by the magnetic beads. The capture efficiency depended on time and sample volume, but did not depend on the length of spacer arm of the biotinylation agent. For cell concentrations of , 105 cfu ml,1, in a 1-ml sample volume, nearly 80,85% recovery of the pathogen was observed after 0·5 h of incubation. For an 11-ml sample containing 104 cfu ml,1, maximum recovery (50% of cells) was achieved only after 2 h incubation. Conclusions:,The detection limit of an ATP-based bioluminescent assay for E. coli O157:H7 was reduced by 1 log cycle after optimization of IMS. The bioluminescent methods could be used for screening and testing the affinity of antibodies or other affinity elements of biosorbents towards live bacterial cells. Significance and Impact of the Study:,Bioluminescent assays provide an easy way to optimize conditions for the capture of bacteria by biosorbents in real time. [source]

    Methyl tert -butyl ether (MTBE)-induced cytotoxicity and oxidative stress in isolated rat spermatogenic cells

    JOURNAL OF APPLIED TOXICOLOGY, Issue 1 2007
    Dongmei Li
    Abstract Methyl tert -butyl ether (MTBE) is a class of synthetic organic chemical. In the USA, MTBE pollution is regarded as a serious environmental problem. The objective of the present study was to investigate the cytotoxic effects and oxidative stress induced by MTBE in isolated rat spermatogenic cells. In cytotoxic experiments, spermatogenic cells isolated from the testes of adult Sprague-Dawley rats by a mechanical procedure without the use of trypsin were incubated with medium alone (control), 0.5, 5, 50 mm MTBE, respectively, for 6, 12 and 18 h. MTT assay, staining with fluorescein diacetate (FDA) and propidium iodide (PI) and flow cytometric analyses were used. In oxidative stress experiments, the spermatogenic cells were incubated with medium alone (control) and with 0.5, 50 ,m, 5 mm MTBE. For 1, 2, 6, 12, 18 h incubation, ROS production was tested using a 2,,7,-dichlorofluorescein diacetate (DCHF-DA) probe; for 1, 3, 6, 12, 18 h incubation, cytosolic superoxide dismutase (SOD) and extracellular SOD (SODEX) activity was assessed; and for 18 h incubation, lipid peroxidation was assessed. The results showed that MTBE at high doses significantly decreased the spermatogenic cell viability and increased plasma membrane damage and the ratio of necrotic cells compared with the control. Assessment of the MTBE-induced oxidative stress revealed that MTBE increased the production of reactive oxygen species (ROS) and enhanced lipid peroxidation. In addition, although SODEX activity increased at a high dose level, cytosolic SOD activity decreased. These results suggest that an increase of MTBE-induced ROS production and an enhancement of membrane lipid peroxidation may play an important role in its cytotoxicity in isolated rat spermatogenic cells. Copyright © 2006 John Wiley & Sons, Ltd. [source]

    Antibacterial effect of silver-zeolite containing root-canal filling material

    JOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 2 2009

    Abstract The aim of this study was to determine the in vitro antibacterial effect of two experimental glass ionomer cements (GICs) on Streptococcus milleri, Staphylococcus aureus, and Enterococcus faecalis after 24 and 48 h incubation by using the agar diffusion inhibitory test. Silver zeolite (SZ) was added at 0.2 and 2% mass fraction concentration to GIC (Endion). The control group was Endion with no SZ. Each of them were prepared to uniform size using a custom-made Teflon mold, and the GIC materials were prepared to form disks (n = 5 per group). The effect of these materials on the growth of three bacteria associated with endodontic infections was determined using the agar diffusion inhibitory test. The amounts of silver ion release from these materials were measured with atomic absorption spectrophotometry at 10 min, 24- and 48-h periods. The pH of samples was measured with a pH-meter at 10 min, 24- and 48-h periods. After the incubation period, the agar plates were evaluated and the degrees of bacterial inhibition were measured in millimeters. A comparison of the mean of the test materials was statistically different in each group of specimens (p < 0.05). Between the two tested materials 2% SZ containing GIC showed the largest zone of inhibition on the agar plates of all the tested strains (p < 0.05). The most inhibition in bacterial growth occurred in E. faecalis. Adding 2% SZ to GIC resulted in a significant increase in the silver release into deionized water. This study demonstrated that GIC had an inhibitory affect on Streptococcus milleri, Staphylococcus aureus, and Enterococcus faecalis and that adding SZ increases that affect proportional to its concentration. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009 [source]

    Composite coating of bonelike apatite particles and collagen fibers on poly L-lactic acid formed through an accelerated biomimetic coprecipitation process

    JOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 2 2006
    Yun Chen
    Abstract Collagen and apatite were coprecipitated as a composite coating on poly L-lactic acid (PLLA) in an accelerated biomimetic process. The incubation solution contained collagen (1 g/L) and simulated body fluid with 5 times inorganic ionic concentrations as human blood plasma. The coating formed on PLLA films and scaffolds after a 24-h incubation was characterized by using energy-dispersive X-ray spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy (SEM). It was shown that the coating contained carbonated bonelike apatite and collagen, which was similar in composition to natural bone. SEM showed a complex composite coating of submicron bonelike apatite particulates combined with collagen fibrils. It is expected that such biocomposite coating may better facilitate cell interaction and osteoconductivity. This work provided an efficient process to obtain bonelike apatite/collagen composite coating, which is potentially useful in bone tissue engineering. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006 [source]

    Regulation of plasminogen activators in human thyroid follicular cells and their relationship to differentiated function

    JOURNAL OF CELLULAR PHYSIOLOGY, Issue 3 2007
    Radhika Susarla
    Human thyroid cells in culture take up and organify 125I when cultured in TSH (acting through cAMP) and insulin. They also secrete urokinase (uPA) and tissue-type (tPA) plasminogen activators (5,100 IU/106cells/day). TSH and insulin both decreased secreted PA activity (PAA), uPA and tPA protein and their mRNAs. Autocrine fibroblast growth factor increased secreted PAA and inhibited thyroid cell 125I uptake. Epidermal growth factor (EGF) and the protein kinase C (PKC) activator, TPA significantly increased PAA and inhibited thyroid differentiated function, (TPA,>,EGF). For TPA, effects were rapid, increased PAA secretion and decreased 125I uptake being seen at 4 h whereas for EGF, a 24 h incubation was required. qRT-PCR showed significantly increased mRNA expression of uPA with lesser effects on tPA. Aprotinin, which inhibits PAA, increased 125I uptake but did not abrogate the effects of TPA and EGF. The MEKK inhibitor, PD98059 partially reversed the effects of EGF and TPA on PAA, and largely reversed the effects of EGF but not TPA on differentiated function. PKC inhibitors bisindoylmaleimide 1, and the specific PKC, inhibitor, LY379196 completely reversed the effects of TPA on 125I uptake and PAA whereas EGF effects were unaffected. TPA inhibited follicle formation and this effect was blocked by LY379196 but not PD98059. We conclude that in thyroid cells, MAPK activation inversely correlates with 125I uptake and directly correlates with PA expression, in contrast to the effects of cAMP. TPA effects on iodide metabolism, dissolution of follicles and uPA synthesis are mediated predominantly through PKC, whereas EGF exerts its effects through MAPK but not PKC,. J. Cell. Physiol. 212:643,654, 2007. © 2007 Wiley-Liss, Inc. [source]

    ASSESSING ABSORBABILITY OF BIOACTIVE COMPONENTS IN ALOE USING IN VITRO DIGESTION MODEL WITH HUMAN INTESTINAL CELL

    JOURNAL OF FOOD BIOCHEMISTRY, Issue 2 2010
    SOON-MI SHIM
    ABSTRACT This study used a simulated in vitro digestion model coupled with caco-2 cell to assess the digestive stability and absorption of aloin, aloe-emodin and aloenin A. Aloenin A and aloe-emodin were stable and entirely recovered during simulated digestion, but 50% of aloin was lost. Approximately 53.2, 7.3 and 28.7% of aloe-emodin, aloenin A and aloin, respectively, was transported into both apical and basolateral compartments after 1 h incubation in caco-2 cell. The involvement of several transporter proteins for aloin and aloenin A was examined. An inhibitor of SGLT1 on apical surface (phloridzin) or that of GLUT2 on basolateral membrane (cytochalasin B) reduced the absorption of aloin by 40 or 60%, respectively, indicating that aloin is likely to be a partial substrate of SGLT1. In the presence of an efflux transporter inhibitor (verapamil), the transport of aloenin A through an intentinal apical membrane increased up to 2.1 times compared with the control (without verapamil). PRACTICAL APPLICATIONS Our results on both digestive stability and intestinal absorption characteristics of bioactive components in aloe could be of helpful information for promoting its bioavailability. The in vitro technique described in this study provides a rapid and cost-effective alternative for predicting bioavailability of biomarkers in aloe functional food. [source]

    WATER ACTIVITY AND THE INACTIVATION OF ENTEROBACTER CLOACAE INOCULATED IN CHOCOLATE LIQUOR AND A MODEL SYSTEM BY PULSED ELECTRIC FIELD TREATMENT

    JOURNAL OF FOOD PROCESSING AND PRESERVATION, Issue 5 2002
    S. MI
    Effects of water activity (aw) on the inactivation of Enterobacter cloacae inoculated in chocolate liquor and in a model system of 0.1% (w/v) peptone water and glycerol by pulsed electric field (PEF) treatment were investigated. An electric field strength of 24.5 kV/cm, a total treatment time of 320 ,s, a pulse duration time of 4 ,s, a pulse delay time of 15 ,s, and a pulse cycle time of 15 s were selected for PEF treatment. The inactivation ofE. cloacae by PEF increased significantly as aw increased (P < 0. 05). As aw of chocolate liquor increased from 0.48 to 0.89, the log reduction of E. cloacae increased from 0.1 to 1.3. The measured temperature change inside the PEF treatment chamber was 0.4C when the log reduction was 1. 3. Similarly, as aw increased from 0. 51 to 0.91 in the model system, the log reduction increased from 0.4 to 1.3. E. cloacae surviving a low aw environment had high resistance to PEF. PEF inactivated E. cloacae in the chocolate liquor with aw of 0.85 by 1 log at O h incubation. However, the log reduction was only 0.1 when PEF treatment was applied to E. cloacae which was incubated for 2 h in the chocolate liquor with aw of 0.85 before PEF treatment. E. cloacae surviving the low aw environment might have resistance not only to the low aw but also to PEF. The resistance to low aw environment may need to be considered when the inactivation of microorganisms by PEF is evaluated. [source]

    Effect of Combining Proteolysis and Lactic Acid Bacterial Fermentation on the Characteristics of Minced Mackerel

    JOURNAL OF FOOD SCIENCE, Issue 3 2005
    Li-Jung Yin
    ABSTRACT: To improve the quality of fish muscle, mackerel muscle protein was hydrolyzed by proteases from Aspergillus oryzae, and then fermented by lactic acid bacteria (LAB). The highest protease activities were obtained from A. oryzae after 72 h incubation at 25°C. Acidic protease activity was much higher than neutral and alkaline proteases. SDS-PAGE indicated the degradation of muscle proteins after 1 or 2 h hydrolysis by A. oryzae proteases at 50°C. During 48 h fermentation by Pediococcus pentosaceus L and S at 37°C, rapid growth of LAB, decline in pH, and suppression in the growth of microflora, Enterobacteriaceae, Staphylococcus, and Pseudomonas, occurred while increases in whiteness, nonprotein nitrogen, sensory quality, and free amino acids were observed. These data suggested that the acceptability of LAB -fermented mackerel hydrolysates could be substantially improved. [source]

    Conversion of Isoflavone Glycosides to Aglycones in SoyLife and Soymeal Using ,-glycosidase

    JOURNAL OF FOOD SCIENCE, Issue 2 2003
    L. Xie
    ABSTRACT: Conversion of isoflavones from glycosides to aglycones in SoyLife and soymeal using varying concentrations of ,-glycosidase, and different pH conditions and temperatures was investigated. The best conditions for the conversion of glycosides to aglycones were pH 5.0, 50 °C, and 5 h incubation with 5 units ,-glycosidase/g Soy Life and 1.5 units/g soymeal. Under these conditions, the amount of genistein, daidzein, and glycitein in Soy Life treated with ,-glycosidase were 4.22, 11.52, and 11.85 ,mol/g compared to untreated controls of 0.26, 0.97, and 4.43 ,mol/g, respectively. In soymeal, the amounts were 3.21, 2.02, and 2.12 ,mol/g compared to untreated controls of 1.23, 1.25, and 1.51 ,mol/g, respectively. Mole percent recovery of genistein was 87% in Soy Life and 80% in soymeal, respectively. [source]

    Hepatocyte dynamics in a three-dimensional rotating bioreactor

    JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, Issue 11 2007
    Mitsuo Miyazawa
    Abstract Background and Aims:, The use of an artificial liver system with extracorporeal circulation or a three-dimensional bioreactor perfused with liquid culture medium inevitably exposes hepatocytes to fluid mechanical stress (MS). The expression of liver-specific hepatocyte functions seems to be modulated by the magnitude of MS. Nonetheless, few studies have focused on the direct effects of MS on hepatocytes. We subjected hepatocytes to MS using an MS loading device and investigated the effects on the cytoskeleton and hepatocyte dynamics inside three-dimensional scaffolds by monitoring the changes in actin fiber, one of the components of the cytoskeleton. We also assessed the influence of MS on specific hepatocyte functions. Methods:, We subjected hepatocytes to MS by a rotating radial flow bioreactor (RRFB) and examined the effects by comparing the MS-loaded culture cells with cells cultured under stationary conditions without MS loading. The hepatocytes (1 × 106/cm3) were seeded on gauze without collagen coating and examined to determine morphological changes after 60 h incubation. Actin filaments in samples from the MS-loaded hepatocyte culture were stained by fluorescein isothiocyanate-labeled phalloidin. Results:, Hepatocyte aggregation was observed in the MS-loaded culture, but not in the unloaded stationary culture. Better albumin products were observed in the MS-loaded group than in the stationary culture group at all measurement points. Actin filaments extended toward the scaffold after the start of MS loading incubation and polymerized around the hepatocytes. The hepatocyte aggregation eventually advanced to the formation of spheroids. Conclusion:, These results suggest that MS-induced polymerization of actin filaments stimulate hepatocyte aggregation and thereby improve hepatocyte-specific function. [source]

    Hypoxia-like effect of Cobalt Chromium alloy micro particles on fibroblasts in vitro

    JOURNAL OF ORTHOPAEDIC RESEARCH, Issue 10 2010
    Bernadette K. Madathil
    Abstract Periprosthetic osteolysis leading to asceptic loosening remains the primary cause of failure of joint replacement. Although many inflammatory cell types have been implicated, the exact pathomechanisms of asceptic loosening have not been delineated. In the present study we have adopted a proteomic approach to elucidate the initial signals that are expressed to particulate material, using an in vitro cell culture system. Human lung fibroblasts MRC-5 were cultured with Cobalt Chromium (CoCr ASTM F-75, 1,7,µm) particles. Cells were harvested after 72,h incubation and total cellular proteins extracted for downstream analysis via 2D Gel Electrophoresis and tandem mass spectrometry using MALDI-TOF-TOF-MS. Thirteen protein spots showed greater than twofold increase, following 72,h incubation of fibroblast with CoCr particles. Four of these proteins were identified by tandem mass spectrometry. These were Annexin II, Pyruvate kinase, Triose phosphate isomerase, and N-myc downstream regulated gene 1 protein. Cobalt is a hypoxia mimicking agent and N-myc downstream regulated gene 1 protein, Triose phosphate isomerase, Pyruvate kinase, and Annexin II are important hypoxia regulated gene products that are found to be over expressed in cellular oxidative stress response. Our data indicates that exposure of fibroblast to CoCr alloy induces the transition of these cells into a hypoxia like state and oxidative stress even in normoxic culture conditions. The study reflects the possibility of the presence of a hypoxic environment in the periprosthetic tissue surrounding metallic implants. Published by Wiley Periodicals, Inc. J Orthop Res 28:1360,1367, 2010 [source]

    Differential apoptotic response of J774 macrophages to alumina and ultra-high-molecular-weight polyethylene particles

    JOURNAL OF ORTHOPAEDIC RESEARCH, Issue 1 2002
    Alain Petit
    We recently identified apoptosis in in vitro wear particle-stimulated macrophages. The recent explosion of interest in apoptosis lies in the fact that it is under positive and negative regulation through evolutionary conserved biochemical pathways. It may also be possible to modulate macrophage apoptosis in the treatment of periprosthetic osteolysis. The purpose of this study was to compare the macrophage response to identically sized particles of alumina ceramic (Al2O3) and ultra-high-molecular-weight polyethylene (UHMWPE) in terms of TNF-, release and induction of apoptosis. J774 mouse macrophages were incubated for 0,24 h in the presence of Al2O3 and UHMWPE particles. TNF-, release was measured by ELISA; Poly(ADP-ribose)polymerase (PARP) and caspase-3 expression was analyzed by Western blot; DNA fragmentation (DNA laddering) was visualized on agarose gel containing ethidium bromide. Al2O3 particles induced TNF-, release after 4 h incubation with concentrations reaching 483 and 800 pg/ml after 24 h with 125 and 250 particles/macrophage, respectively (control = 161 pg/ml) (P < 0.05 vs. control). The same concentrations of UHMWPE particles induced a much larger and significant TNF-, release after only 1 h incubation, increasing up to 6250 pg/ml after 24 h (P < 0.05 vs. control). Western blot analysis demonstrated that the active caspase-3 fragment (17 kDa) and the proteolytic PARP fragment (85 kDa) were expressed after 2 h incubation with 125 and 250 Al2O3 particles/macrophage. The active caspase-3 and the PARP fragment had lower expression and appeared after a longer incubation time (8 h) with 125 and 250 UHMWPE particles/macrophage. Finally, DNA fragmentation (DNA laddering) was observed after 16 h with 125 and 250 particles of Al2O3 per macrophage whereas no laddering was induced by UHMWPE particles even after 24 h incubation. This study shows that although both Al2O3 and UHMWPE particles induce TNF-, release, this stimulation was much greater (8,10 times higher) with UHMWPE than A12O3 (P < 0.05 vs. control). As well, the induction of apoptosis, as measured by activation of caspase-3, PARP cleavage and DNA laddering, is different for these two particles, being faster and more important with Al2O3 than UHMWPE. We hypothesize that the ability of Al2O3 to induce macrophage apoptosis may explain the lower TNF-, release observed with these particles and explain the differences seen in osteolysis patterns of ceramic,ceramic (CC) vs. metal,polyethylene (Mpe) articulations. In conclusion, apoptosis may be a major internal mechanism to decrease macrophage activity and may be a desired therapeutic endpoint. The identification of an apoptosis-related pathway in the macrophage response to ceramic particles provides crucial data for a rational approach in the treatment and/or prevention of periprosthetic osteolysis. © 2002 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved. [source]

    Physico-enzymatic production of monoacylglycerols enriched with very-long-chain polyunsaturated fatty acids

    JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 2 2008
    Ratchapol Pawongrat
    Abstract BACKGROUND: Monoacylglycerols (MAG) containing polyunsaturated fatty acids (PUFA), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), have interesting applications. The enzymatic processing of such MAG directly from fish oils is highly interesting, integrating the processing of MAG and concentration of EPA and DHA. The aim of this study was to develop an efficient enzymatic glycerolysis system together with physical fractionation for the production of PUFA-MAG from tuna oil. RESULTS: Novozym 435 was eventually selected after evaluation together with immobilized lipase AK in a tertiary alcohol-based system. A further evaluation of solvent mixtures involving tertiary alcohols was made, taking ease of operation into consideration. It turned out that a number of mixtures gave a similar performance to that of tert -butanol (TB). Basic reaction parameters were thoroughly evaluated. In the batch reaction system with TB as solvent, the recommended conditions were: glycerol/tuna oil 4:1 (mol/mol), TB/tuna oil 2:1 (wt/wt), 15 wt% Novozym 435, and temperature 40 °C. Under these conditions, the yield of MAG was up to 90% after 3 h incubation. Crude MAG from the production was fractionated to produce MAG with higher EPA and DHA content. Using acetone as solvent at 0 °C led to ca 50% yield of MAG but contained EPA and DHA up to 71% in comparison with ca 30% in tuna oil. CONCLUSION: Potentially practical process steps have been developed for the production of MAG containing a high content of EPA and DHA from natural fish oils with high efficiency and simplicity. Copyright © 2007 Society of Chemical Industry [source]

    Polyphenol oxidase activity in grass and its effect on plant-mediated lipolysis and proteolysis of Dactylis glomerata (cocksfoot) in a simulated rumen environment

    JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 10 2006
    Michael RF Lee
    Abstract Little is known about the level or activity of polyphenol oxidase (PPO) in grasses and its potential impact on proteolysis and lipolysis. Six grass species were initially screened for PPO activity (740.6, 291.9, 213.6, 119.0, 16.3 and 6.5 U g,1 fresh weight (FW) for cocksfoot, hybrid ryegrass, Italian ryegrass, perennial ryegrass, timothy and tall fescue respectively). Cocksfoot, which expressed the highest activity, was then used to determine the effect of PPO on plant-mediated proteolysis and lipolysis in a simulated rumen environment. Sourced cocksfoot was macerated and incubated in an antibiotic-containing anaerobic medium with or without ascorbate to deactivate PPO in the dark at 39 °C over five time points. At each time point (0, 1, 2, 6 and 24 h), six replicate samples were destructively harvested; three of the replicates were used for lipid analysis and the other three for protein, free amino acid and bound phenol determination. Characterisation of the herbage showed PPO activities of 649.6 and 0 U g,1 FW, which were reflected in the extent of phenol (derived from quinones) binding to protein after 24 h of incubation, namely 65.1 and 29.6 mg bound phenol g,1 protein (P < 0.001) for cocksfoot and cocksfoot + ascorbate respectively. Proteolysis, measured as free amino acids released into the incubation buffer, was significantly reduced (P < 0.001) with increasing PPO activity, with values after the 24 h incubation of 0.03 and 0.07 mmol L,1 g,1 FW for cocksfoot and cocksfoot + ascorbate respectively. Lipolysis, measured as the proportional decline in the membrane lipid polar fraction, was likewise reduced (P < 0.001) with increasing PPO activity, with values after the 24 h incubation of 0.43 and 0.65 for cocksfoot and cocksfoot + ascorbate respectively. Changes that occurred in protein and the lipid fractions (polar fraction, monoacylglycerol + diacylglycerol, triacylglycerol and free fatty acids) during the incubations are also reported and discussed. These results support the selection of forages high in PPO activity to reduce protein and lipid losses in silo and potentially in the rumen. Copyright © 2006 Society of Chemical Industry [source]

    Metabolism of isometamidium in hepatocytes isolated from control and inducer-treated rats

    JOURNAL OF VETERINARY PHARMACOLOGY & THERAPEUTICS, Issue 6 2006
    I. BOIBESSOT
    Little is known about the metabolism and mechanism of action of the trypanocide, isometamidium (ISM), the major drug used for prophylaxis of trypanosomiasis. We have investigated its metabolism and distribution in isolated rat hepatocytes using liquid chromatography-mass spectrometry and confocal laser scanning microscopy (CLSM). Two putative metabolites were formed, which were proposed to be a mono-acetyl derivative and an oxidized metabolite (SII). This is the first demonstration of the hepatic metabolism of ISM, as previous in vivo studies were hampered by dose-limiting toxicity and insensitive analytical methods. The intrinsic fluorescence of the drug enabled its intracellular uptake to be followed by CLSM. It is taken up rapidly into the nucleolus, nuclear membrane and endoplasmic reticulum within 5 min, and retained in the nucleus for at least 24 h. Persistent binding of ISM to cellular macromolecules may contribute to its prophylactic effect in vivo. Pretreatment of rats with 3-methylcholanthrene, phenobarbitone (PB) or the widely used pyrethroid pesticide, deltamethrin, resulted in an increase in metabolism of ISM to the proposed SII after 1 h incubation with hepatocytes. 3-methylcholanthrene was the most potent inducer, causing a maximal 19.5-fold induction of SII formation after exposure of hepatocytes to ISM for 1 h compared with formation by control hepatocytes. In comparison, at the 1 h timepoint deltamethrin pre-treatment caused a 10.2-fold induction, and PB only 8.2 fold. [source]

    Moxidectin and ivermectin metabolic stability in sheep ruminal and abomasal contents

    JOURNAL OF VETERINARY PHARMACOLOGY & THERAPEUTICS, Issue 5 2005
    A. LIFSCHITZ
    The oral administration of macrocyclic lactones to sheep leads to poorer efficacy and shorter persistence of the antiparasitic activity compared to the subcutaneous treatment. Gastrointestinal biotransformation occurring after oral treatment to ruminant species has been considered as a possible cause of the differences observed between routes of administration. The current work was addressed to evaluate on a comparative basis the in vitro metabolism of moxidectin (MXD) and ivermectin (IVM) in sheep ruminal and abomasal contents. Both compounds were incubated under anaerobic conditions during 2, 6 and 24 h in ruminal and abomasal contents collected from untreated adult sheep. Drug concentrations were measured by high-performance liquid chromatography with fluorescence detection after sample clean up and solid phase extraction. Neither MXD nor IVM suffered metabolic conversion and/or chemical degradation after 24-h incubation in ruminal and abomasal contents collected from adult sheep. Unchanged MXD and IVM parent compounds represented between 95.5 and 100% of the total drug recovered in the ruminal and abomasal incubation mixtures compared with those measured in inactive control incubations. The partition of both molecules between the solid and fluid phases of both sheep digestive contents was assessed. MXD and IVM were extensively bound (>90%) to the solid material of both ruminal and abomasal contents collected from sheep fed on lucerne hay. The results reported here confirm the extensive degree of association to the solid digestive material and demonstrates a high chemical stability without evident metabolism and/or degradation for both MXD and IVM in ruminal and abomasal contents. [source]

    Pharmacokinetics and pharmacokinetic/pharmacodynamic integration of marbofloxacin in calf serum, exudate and transudate

    JOURNAL OF VETERINARY PHARMACOLOGY & THERAPEUTICS, Issue 3 2002
    F. SHOJAEE ALIABADI
    Aliabadi, F. S., Lees, P. Pharmacokinetics and pharmacokinetic/pharmacodynamic integration of marbofloxacin in calf serum, exudate and transudate. J. vet. Pharmacol. Therap.25, 161,174. Marbofloxacin is a fluoroquinolone antimicrobial drug used in cattle for the treatment of respiratory infections. In this investigation the pharmacokinetics (PK) of marbofloxacin were determined after intravenous and intramuscular dosing at a dosage of 2 mg/kg. In addition the ex vivo pharmacodynamics (PD) of the drug were determined in serum and three types of tissue cage fluid (transudate, inflammatory exudate generated by carrageenan and exudate generated by lipopolysaccharide). Marbofloxacin PK was characterized by a high volume of distribution after dosing by both routes (1.28 L/kg intravenous and 1.25 L/kg intramuscular). Corresponding area under the concentration,time curve (AUC) and elimination half-life (t½el) values were 9.99 and 10.11 ,g h/mL and 4.23 and 4.33 h, respectively. Values of AUC for carrageenan-induced exudate, lipopolysaccharide-induced exudate and transudate were, respectively, 8.28, 7.83 and 7.75 ,g h/mL after intravenous and 8.84, 8.53 and 8.52 ,g h/mL after intramuscular dosing. Maximum concentration (Cmax) values were similar for the three tissue cage fluids after intravenous and intramuscular dosing. For in vivo PK data values of AUC: minimum inhibitory concentration (MIC) (AUIC) ratio for serum were 250 and 253, respectively, after intravenous and intramuscular dosing of marbofloxacin against a pathogenic strain of Mannheimia haemolytica (MIC=0.04 ,g/mL). For all tissue cage fluids AUIC values were >194 and >213 after intravenous and intramuscular dosing, and Cmax/MIC ratios were 9 or greater, indicating a likely high level of effectiveness in clinical infections caused by M. haemolytica of MIC 0.04 ,g/mL or less. This was confirmed by both in vitro (serum) and ex vivo (serum, exudate and transudate) measurements, which demonstrated a concentration-dependent killing profile for marbofloxacin against M. haemolytica. Ex vivo, after 24-h incubation, virtually all bacteria were killed (<10 cfu/mL) in all samples collected up to 9 h (serum), 24 h (carrageenan-induced exudate and transudate) and 36 h (lipopolysaccharide-induced exudate). Application of the sigmoid Emax equation to the ex vivo antibacterial data provided, for serum, AUIC24 h values of 37.1 for bacteriostasis, 46.3 for bactericidal activity and 119.6 for elimination of bacteria. These data may be used as a rational basis for setting dosing schedules which optimize clinical efficacy and minimize the opportunities for emergence of resistant organisms. [source]

    A Novel Micellar PEGylated Hyperbranched Polyester as a Prospective Drug Delivery System for Paclitaxel

    MACROMOLECULAR BIOSCIENCE, Issue 9 2008
    Christina Kontoyianni
    Abstract A hyperbranched aliphatic polyester has been functionalized with PEG chains to afford a novel water-soluble BH40-PEG polymer which exhibits unimolecular micellar properties, and is therefore appropriate for application as a drug-delivery system. The solubility of the anticancer drug paclitaxel was enhanced by a factor of 35, 110, 230, and 355 in aqueous solutions of BH40-PEG of 10, 30, 60, and 90 mg,·,mL,1, respectively. More than 50% of the drug is released at a steady rate and release is almost complete within 10 h. The toxicity of BH40-PEG was assessed in vitro with A549 human lung carcinoma cells and found to be nontoxic for 3 h incubation up to a 1.75 mg,·,mL,1 concentration while LD50 was 3.5 mg,·,mL,1. Finally, it was efficiently internalized in cells, primarily in the absence of foetal bovine serum, while confocal microscopy revealed the preferential localization of the compound in cell nuclei. [source]

    Nematicidal activity of anion transport blockers against Meloidogyne incognita, Caenorhabditis elegans and Heterorhabditis bacteriophora

    PEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 6 2008
    Dhana Raj Boina
    Abstract BACKGROUND: Because methyl bromide has been phased out as a soil sterilant, new nematicides are urgently needed. Four different chemical classes of organic acids acting as anion transport (AT) blockers were tested against a free-living nematode, Caenorhabditis elegans Maupas, a plant-parasitic nematode, Meloidogyne incognita (Kofoid and White) Chitwood, and an entomopathogenic nematode, Heterorhabditis bacteriophora Poinar, in toxicity bioassays. The materials tested were DIDS (4,4,-diisothiocyanatostilbene-2,2,-disulfonic acid), 9-AC (anthracene-9-carboxylic acid), NPPB [5-nitro-2-(3-phenylpropylamino)benzoic acid] and IAA-94 (indanyloxyacetic acid). RESULTS: All the compounds showed slowly developing nematicidal activity against second-stage juveniles of M. incognita and adults of C. elegans, but not against H. bacteriophora infective-stage juveniles. The LC50 values of these compounds were < 50 mg L,1 after 48 and 72 h incubation, while at 168 h incubation the LC50 values were < 10 mg L,1 for both sensitive species. Across both species and time, the LC50 values generally differed no more than twofold among the four compounds tested in this study. In contrast, none of the compounds (200 mg L,1) caused more than control mortality to H. bacteriophora, even after 168 h of incubation. CONCLUSION: These compounds are potential leads for commercial nematicides. The insensitivity to H. bacteriophora is consistent with the natural exposure of this nematode to DST (3,5-dihydroxy-4-isopropylstilbene), a stilbene produced by its symbiotic bacterium. Based on the known activity of the compounds used in this study, it is suggested that anion transporters form the probable target sites for DIDS, 9-AC, NPPB and IAA-94 in nematodes. Copyright © 2008 Society of Chemical Industry [source]