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Vitro Tests (vitro + test)
Selected AbstractsSteroidogenic gene expression in H295R cells and the human adrenal gland: adrenotoxic effects of lindane in vitroJOURNAL OF APPLIED TOXICOLOGY, Issue 6 2006Agneta Oskarsson Abstract The focus on the refinement, reduction and replacement of animal use in toxicity testing requires the development of cell-based systems that mimic the effects of xenobiotics in human tissues. The human adrenocortical carcinoma cell line, H295R, has been proposed as a model for studies on adrenal steroidogenesis and its disruption. In this study, expression profiles for nine adrenal steroidogenic genes were characterized in H295R cells using real-time RT-PCR. Treatment with forskolin increased cortisol secretion and stimulated transcription of all the steroidogenic genes except SULT2A1. The transcript profile from H295R cells in the presence and absence of forskolin was compared with the transcript profile from human adrenal glands. The gene expression pattern observed in the forskolin-treated H295R cells was more similar to that in the human adrenal gland, than the expression pattern in untreated cells. To examine H295R cells as a possible in vitro system for the assessment of adrenal disruption using molecular endpoints, the insecticide lindane (, -hexachlorocyclohexane) was used. In vivo, lindane has been shown to inhibit testicular, ovarian and adrenal steroidogenesis. It was demonstrated that lindane reduced cortisol secretion, downregulated the expression of a subset of the genes encoding steroidogenic enzymes and repressed transcriptional activation of the steroidogenic acute regulatory protein (StAR) gene promoter. Thus the H295R cell line provides a good in vitro system for the analysis of the human adrenal steroidogenic pathway at the level of hormone production and gene expression. This in vitro test can be used for the rapid detection of adrenal endocrine disruption and as a tool for mechanistic studies. Copyright © 2006 John Wiley & Sons, Ltd. [source] A New Stent Design for the Treatment of True Bifurcation Lesions: H-Side Branch StentsJOURNAL OF INTERVENTIONAL CARDIOLOGY, Issue 1 2010MYEONG-KI HONG M.D. Background:There has been much debate for the adequate treatment strategies for true bifurcation lesions. The purpose of this study is to introduce and test a novel stent design for the treatment of true bifurcation lesions. Methods:This side branch stent is composed of three parts: proximal, connecting, and distal parts. The distal part for the side branch vessel has a slope-side stent margin for circumferential coverage of the ostium and one radio-opaque marker for targeting the carina. The proximal part with two radio-opaque markers operates for safe stent delivery and useful guidance for a more precise placement of the distal part on the side branch ostium. Results of the in vitro test in the acrylic resin-made bifurcation phantom model were evaluated with microcomputer tomography. Animal experiments with this new stent platform were also performed in five pigs. Results:In vitro test and microcomputer tomography showed complete coverage of the side branch ostium circumferentially with stent struts, and the absence of stent struts in the main vessel above the side branch ostium level. This side branch stents were successfully deployed in all 5 pigs. The results of animal experiments were also similar to those of in vitro tests. Conclusions:In vivo and vitro tests demonstrated the effective modality of this side branch stent for the treatment of true bifurcation lesions. (J Interven Cardiol 2010;23:54,59) [source] EAACI/GA2LEN guideline: aspirin provocation tests for diagnosis of aspirin hypersensitivityALLERGY, Issue 10 2007E. Ni, ankowska-Mogilnicka Abstract:, Aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most common causes of adverse drug reactions. Majority of them are of the hypersensitivity type. The two frequent clinical presentations of aspirin hypersensitivity are: aspirin-induced bronchial asthma/rhinosinusitis (AIA/R) and aspirin-induced urticaria/angioedema (AIU). The decisive diagnosis is based on provocation tests with aspirin, as the in vitro test does not hold diagnostic value as yet. Detailed protocols of oral, bronchial and nasal aspirin provocation tests are presented. Indications, contraindications for the tests, the rules of drug withdrawal and equipment are reviewed. Patient supervision and interpretations of the tests are proposed. [source] Analysis of basophil activation by flow cytometry in pediatric house dust mite allergyPEDIATRIC ALLERGY AND IMMUNOLOGY, Issue 4 2008Miguel González-Muñoz Detection of allergen-induced basophil activation by flow cytometry has been shown to be a useful tool for allergy diagnosis. The aim of this study was to assess the potential of this technique for the diagnosis of pediatric house dust mite allergy. Quantification of total and specific IgE and basophil activation test were performed to evaluate mite allergic (n = 24), atopic (n = 23), and non-allergic children (n = 9). Allergen-induced basophil activation was detected as a CD63-upregulation. Receiver operating characteristics (ROC) curve analysis was performed to calculate the optimal cut-off value of activated basophils discriminating mite allergic and non-allergic children. ROC curve analysis yielded a threshold value of 18% activated basophils when mite-sensitized and atopic children were studied [area under the curve (AUC) = 0.99, 95% confidence interval (CI) = 0.97,1.01, p < 0.001] with a sensitivity and specificity of 96% for 16 ,g/ml mite extract. Analysis of the data obtained with 1.6 ,g/ml mite extract defined a cut-off value of 8% activated basophils (AUC = 0.96, 95% CI = 0.91,1.01; p < 0.001) with a sensitivity of 82% and specificity of 100%. Comparison between mite allergic and non-allergic children produced a cut-off of 8% activated basophils (AUC = 1.0) with 16 ,g/ml allergen extract and a sensitivity and specificity of 100%. The same threshold and specificity values were obtained with 1.6 ,g/ml extract (AUC = 97%, 95% CI = 0.92,1.02; p < 0.001) but sensitivity decreased to 83%. Two atopic children showed negative skin prick and basophil activation tests and high specific IgE (>43 kU/l) values for Dermatophagoides pteronyssinus allergen. They also showed positive prick (wheal diameter >1.0 cm) and basophil activation (>87%) tests and high specific IgE (>100 kU/l) with shrimp allergen. Shrimp sensitization was demonstrated by high levels of Pen a 1-specific IgE (>100 kU/l). Cross-reactivity between mite and shrimp was confirmed by fluorescence enzyme immunoassay (FEIA-CAP) inhibition study in these two cases. This study demonstrated that the analysis of allergen-induced CD63 upregulation by flow cytometry is a reliable tool for diagnosis of mite allergy in pediatric patients, with sensitivity similar to routine diagnostic tests and a higher specificity. Furthermore, this method can provide additional information in case of disagreement between in vivo and in vitro test results. [source] Characterisation of QoI-resistant field isolates of Botrytis cinerea from citrus and strawberryPEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 8 2009Hideo Ishii Abstract BACKGROUND: In 2004, field isolates of Botrytis cinerea Pers. ex Fr., resistant to strobilurin fungicides (QoIs), were first found in commercial citrus orchards in Wakayama Prefecture, Japan. Subsequently, QoI-resistant isolates of this fungus were also detected in plastic strawberry greenhouses in Saga, Ibaraki and Chiba prefectures, Japan. Biological and molecular characterisation of resistant isolates was conducted in this study. RESULTS: QoI-resistant isolates of B. cinerea grew well on PDA plates containing kresoxim-methyl or azoxystrobin at 1 mg L,1, supplemented with 1 mM of n -propyl gallate, an inhibitor of alternative oxidase, whereas the growth of sensitive isolates was strongly suppressed. Results from this in vitro test were in good agreement with those of fungus inoculation tests in vivo. In resistant isolates, the mutation at amino acid position 143 of the cytochrome b gene, known to be the cause of high QoI resistance in various fungal pathogens, was found, but only occasionally. The heteroplasmy of cytochrome b gene was confirmed, and the wild-type sequence often present in the majority of resistant isolates, indicating that the proportion of mutated cytochrome b gene was very low. CONCLUSION: The conventional RFLP and sequence analyses of PCR-amplified cytochrome b gene are insufficient for molecular identification of QoI resistance in B. cinerea. Copyright © 2009 Society of Chemical Industry [source] Synthesis, Crystal Structure and in vitro Antitumor Activity of Di- n -butyltin p -[N, N -Bis(2-chloroethyl)amino]benzoatesCHINESE JOURNAL OF CHEMISTRY, Issue 12 2005Zhong-Wei Zhang Abstract Di- n -butyltin oxide reacted with p- [N,N -bis(2-chloroethyl)amino]benzoic acid to yield the compounds {{4-[(ClCH2CH2)2N]C6H4COOSnBu2}2O}2 (1) and {4-[(ClCH2CH2)2N]C6H4COO}2SnBu2 (2), which have been characterized by IR and 1H NMR spectra. The X-ray diffractional studies of 1 reveal the structure of the molecule to be a dimer, in which the two Bu2Sn groups were linked via two bridging oxygen atoms to form a central Bu4Sn2O2 unit. And the tin atom adopts two carbons from two n -butyl groups and three oxygen atoms from the acid and the bridging oxygen. In vitro test showed compound 1 to exhibit high cytotoxicity against P388 and HL-60 cell lines. [source] Evaluation of effects of rofecoxib on platelet function in an in vitro model of thrombosis with circulating human bloodEUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 4 2004M. R. Hernandez Abstract Background, Cyclooxygenase (COX)-2-selective non-steroidal anti-inflammatory drugs have been used for anti-inflammatory therapy. However, it has also been described that they may increase risk of cardiovascular events. Objectives, To study the effects of COX2 inhibitor rofecoxib on platelet function using in vitro tests. Results were compared with those obtained in a parallel experiment with acetyl salicylic acid (ASA). Methods, Studies of platelet aggregation, using different agonists, were performed by a turbidimetric method. Adhesive and cohesive function of platelets were analyzed by perfusion techniques, treated blood was exposed to thrombogenic surfaces and platelet interaction was morphometrically evaluated. Results, Twenty-five µM of rofecoxib induced a prolonged lag time and a reduction in the percentage of aggregation when arachidonic acid, ADP or collagen were used as agonists. In perfusion studies with parallel chamber rofecoxib 50 µM and ASA 500 µM reduced overall platelet interaction with the collagen surface (17·4 ± 3·7, P < 0·05; vs. 32·1 ± 2·6%P < 0·05 and 17·9 ± 2·4, vs. 31·9 ± 3·24, P < 0·05, respectively). In studies performed on annular chambers, 25 µM of rofecoxib reduced platelet interaction; values of the thrombus and covered surface were 17·4 ± 4·5%; P < 0·05 and 21·1 ± 4·1%; P < 0·05, respectively, vs. 30·4 ± 7·5% and 33·5 ± 6·5 in the control. ASA did also impair thrombus formation but differences did not reach the levels of statistical significance. Moreover, rofecoxib but not ASA reduced significantly thrombus height and thrombus area (7·4 ± 0·5 µM; P < 0·005 and 96·0 ± 21·2 µM2; P < 0·05 vs. control 11·2 ± 0·9 µM and 220·0 ± 47·7µM2, respectively). Conclusion, We conclude that under our experimental conditions, rofecoxib diminished platelet aggregation induced by different agonists and inhibited platelet-mediated thrombogenesis in an in vitro model of thrombosis. [source] Assessment of CD8 involvement in T,cell clone avidity by direct measurement of HLA-A2/Mage3 complex density using a high-affinity TCR like monoclonal antibodyEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 10 2005Karine Bernardeau Abstract Peptide affinity for MHC molecules determines the number of MHC/peptide complexes stabilized at the cell surface in in vitro tests or in vaccination protocols. We isolated a high affinity monoclonal antibody specific for the HLA-A2/Mage3 complex that enables an equilibrium binding assay to be performed on T2 cell line loaded with a range of Mage3 peptides. Binding of Mage3 to the HLA-A2 molecule can be modeled by a standard receptor-ligand interaction characterized by an affinity constant. This model enables the measurement of the affinity of other immunogenic peptides for HLA-A2 by a competition test and the calculation of the density of complexes stabilized at the T2 cell surface for all peptide concentrations. Quantification of the HLA-A2/Mage3 complexes at target cell surfaces was used to estimate the number of complexes required to reach cytotoxicity ED50 of human T,cell clones sorted from an unprimed repertoire. We confirm with this antibody the direct relationship between clone avidity and TCR affinity, and the moderate contribution of the CD8 co-receptor in the reinforcement of TCR-MHC/peptide contact. Nevertheless, CD8 plays a critical role in the amplification of the specific signal to establish an efficient T,cell response at low specific complex densities found in physiological situations. [source] The New Metabolite (S)-Cinnamoylphosphoramide from Streptomyces sp. and Its Total SynthesisEUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 30 2008Melanie Quitschau Abstract The tunicate-associated strain Streptomyces sp. JP90 produces the unprecedented metabolite cinnamoylphosphoramide (1) among several other compounds. Structure elucidation was accomplished by NMR spectroscopic studies and efficient total synthesis. The absolute configuration at phosphorus was determined by synthesis of both enantiomers of 1 performing a resolution of the corresponding diastereomeric phosphoramides of L -phenylalanine ethyl ester. Unusual cinnamic acid derivative 1 represents the first bacterial organophosphoramide. As it matches the Schrader's formula for insecticidal organophosphates, its biological activity was investigated. A weak inhibition of acetylcholinesterase was observed in in vitro tests, and water-soluble analogues of 1 were prepared. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008) [source] Study of the Reaction of 3-Formylrifamycin SV with Gaseous Ammonia and Acetone,EUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 18 2006Krzysztof Bujnowski Abstract In the second stage of our study concerning the search for new antibacterial rifamycin antibiotics, reaction of the aldehydes 3-formylrifamycin SV (1) and 25- O -deacetyl-3-formylrifamycin SV with ammonia and acetone has been investigated. A new synthetic method for the preparation of a new group of rifamycin derivatives with a cyclic substituent at C-3 having a 4-piperidone structure, represented by compounds 6a, 6b, and 7a, has been developed. The structures of the compounds and a reaction mechanism have been proposed on the basis of mass spectrometry results as well as 1D and 2D 1H and 13C NMR analysis. The results of the in vitro tests confirm the antituberculous activity of the synthesized compounds. Furthermore, 6a, which is isolated in good yield, is a promising substrate for a new class of rifamycin derivatives. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2006) [source] The mechanisms of resistance to antimalarial drugs in Plasmodium falciparumFUNDAMENTAL & CLINICAL PHARMACOLOGY, Issue 2 2003Jacques Le Bras Abstract Drug-resistant malaria is primarily caused by Plasmodium falciparum, a species highly prevalent in tropical Africa, the Amazon region and South-east Asia. It causes severe fever or anaemia that leads to more than a million deaths each year. The emergence of chloroquine resistance has been associated with a dramatic increase in malaria mortality among inhabitants of some endemic regions. The rationale for chemoprophylaxis is weakening as multiple-drug resistance develops against well-tolerated drugs. Plasmodium falciparum drug-resistant malaria originates from chromosome mutations. Analysis by molecular, genetic and biochemical approaches has shown that (i) impaired chloroquine uptake by the parasite vacuole is a common characteristic of resistant strains, and this phenotype is correlated with mutations of the Pfmdr1, Pfcg2 and Pfcrt genes; (ii) one to four point mutations of dihydrofolate reductase (DHFR), the enzyme target of antifolates (pyrimethamine and proguanil) produce a moderate to high level of resistance to these drugs; (iii) the mechanism of resistance to sulfonamides and sulfones involves mutations of dihydropteroate synthase (DHPS), their enzyme target; (iv) treatment with sulphadoxine,pyrimethamine selects for DHFR variants Ile(51), Arg(59), and Asn(108) and for DHPS variants Ser(436), Gly(437), and Glu(540); (v) clones that were resistant to some traditional antimalarial agents acquire resistance to new ones at a high frequency (accelerated resistance to multiple drugs, ARMD). The mechanisms of resistance for amino-alcohols (quinine, mefloquine and halofantrine) are still unclear. Epidemiological studies have established that the frequency of chloroquine resistant mutants varies among isolated parasite populations, while resistance to antifolates is highly prevalent in most malarial endemic countries. Established and strong drug pressure combined with low antiparasitic immunity probably explains the multidrug-resistance encountered in the forests of South-east Asia and South America. In Africa, frequent genetic recombinations in Plasmodium originate from a high level of malaria transmission, and falciparum chloroquine-resistant prevalence seems to stabilize at the same level as chloroquine-sensitive malaria. Nevertheless, resistance levels may differ according to place and time. In vivo and in vitro tests do not provide an adequate accurate map of resistance. Biochemical tools at a low cost are urgently needed for prospective monitoring of resistance. [source] Prediction of hemodialysis sorbent cartridge urea nitrogen capacity and sodium release from in vitro testsHEMODIALYSIS INTERNATIONAL, Issue 2 2008Benjamin P. ROSENBAUM Abstract In sorbent-based hemodialysis, factors limiting a treatment session are urea conversion capacity and sodium release from the cartridge. In vitro experiments were performed to model typical treatment scenarios using various dialyzers and 4 types of SORBÔ sorbent cartridges. The experiments were continued to the point of column saturation with ammonium. The urea nitrogen removed and amount of sodium released in each trial were analyzed in a multi-variable regression against several variables: amount of zirconium phosphate (ZrP), dialysate flow rate (DFR), simulated blood flow rate (BFR), simulated patient whole-body fluid volume (V), initial simulated patient urea concentration (BUNi), dialyzer area permeability (KoA) product, initial dialysate sodium and bicarbonate (HCO3i) concentrations, initial simulated patient sodium (Nai), pH of ZrP, creatinine, breakthrough time, and average urea nitrogen concentration in dialysate. The urea nitrogen capacity (UNC) of various new SORBÔ columns is positively related to ZrP, BFR, V, BUNi, and ZrP pH and negatively to DFR with an R2adjusted=0.990. Two models are described for sodium release. The first model is related positively to DFR and V and negatively to ZrP, KoA product, and dialysate HCO3i with an R2adjusted=0.584. The second model incorporates knowledge of initial simulated patient sodium (negative relationship) and urea levels (negative relationship) in addition to the parameters in the first model with an R2adjusted=0.786. These mathematical models should allow for prediction of patient sodium profiles and the time of column urea saturation based on simple inputs relating to patient chemistries and the dialysis treatment. [source] Protective effect of non-mitogenic human acidic fibroblast growth factor on hepatocyte injuryHEPATOLOGY RESEARCH, Issue 10 2007Hua Xu Aim:, To study whether non-mitogenic human acidic fibroblast growth factor (nm-haFGF) has protective effects on H2O2 -induced hepatocyte injury in vitro and CCl4 -induced hepatocyte injury in vivo. Methods:, (i) HL-7702 hepatocytes were incubated with different concentrations of nm-haFGF for 12 h, and then the activity of lactate dehydrogenase (LDH) in culture medium was detected, and genomic DNA electrophoresis analysis was observed after being exposed to H2O2 (8 mmol/L) for 4 h. Proximately, apoptotic rates and protein expressions of Bcl-2 and Bax of HL-7702 cell were detected after being exposed to H2O2 (0.2 mmol/L) for 20 h. (ii) Being injected intraperitoneally with nm-haFGF, mice were treated with CCl4 intraperitoneally to induce hepatic injury. Twenty-four hours later, serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured and histopathologic changes were evaluated. Results:, (i) In vitro tests: LDH activities and apoptotic rates decreased, the protein expression of Bcl-2 increased and Baxdecreased in nm-haFGF-treated groups at the concentrations of 100 150 and 200 ng/mL, compared with that in the model control group, which was treated with H2O2 alone. The genomic DNA remained nearly intact at the concentrations of 150 and 200 ng/mL. (ii) In vivo tests: serum ALT and AST in nm-haFGF-treated groups (10 ,g/kg and 20 ,g/kg) were much lower as compared to the model control group, which was treated with CCl4 alone. Histological examination showed that nm-haFGF markedly ameliorated hepatocytes vacuolation, cloudy swelling and inflammatory cells infiltration induced by CCl4. Conclusion:, nm-haFGF had protective effects against H2O2 -induced hepatocyte injury in vitro and CCl4 -induced acute liver injury in vivo. [source] Sperm function tests and fertilityINTERNATIONAL JOURNAL OF ANDROLOGY, Issue 1 2006R. J. Aitken Summary Traditionally, the diagnosis of male infertility has depended upon a descriptive evaluation of human semen with emphasis on the number of spermatozoa that are present in the ejaculate, their motility and their morphology. The fundamental tenet underlying this approach is that male fertility can be defined by reference to a threshold concentration of motile, morphologically normal spermatozoa that must be exceeded in order to achieve conception. Many independent studies have demonstrated that this fundamental concept is flawed and, in reality, it is not so much the absolute number of spermatozoa that determines fertility, but their functional competence. In the light of this conclusion, a range of in vitro tests have been developed to monitor various aspects of sperm function including their potential for movement, cervical mucus penetration, capacitation, zona recognition, the acrosome reaction and sperm,oocyte fusion. Such functional assays have been found to predict the fertilizing capacity of human spermatozoa in vitro and in vivo with some accuracy. Recent developments in this field include the introduction of tests to assess the degree to which human spermatozoa have suffered oxidative stress as well as the integrity of their nuclear and mitochondrial DNA. Such assessments not only yield information on the fertilizing capacity of human spermatozoa but also their ability to support normal embryonic development. [source] Cell Proliferation of Human Fibroblasts on Alumina and Hydroxyapatite-Based Ceramics with Different Surface Treatments,INTERNATIONAL JOURNAL OF APPLIED CERAMIC TECHNOLOGY, Issue 2 2010Juliana Marchi Biocompatibility is an important characteristic of dental implant material, and in vitro tests are required to elucidate the interaction between these materials and human tissues. Cell proliferation assays were done with fibroblasts plated on the surface of alumina and hydroxyapatite sintered samples, each with a different surface treatment (sintered, rectified, or polished). After 1, 2, and three days, the samples were prepared for scanning electron microscopy observations. The data were compared by analysis of variance followed by Tukey's test. It was concluded that neither the hydroxyapatite or alumina substrate is cytotoxic, and hydroxyapatite is more biocompatible than alumina. [source] Anti-microbial hand washes for domestic use , their effectiveness in vitro and in normal useINTERNATIONAL JOURNAL OF CONSUMER STUDIES, Issue 3 2001Kay Sharp Abstract The killing or removal of microbes from the hands is a critical factor in food safety as many studies have shown the hands to be both an important source of microbes and powerful agents of cross-contamination in hospital and domestic situations. In response to this concern, a number of novel hand-washing products have appeared on the market. These products contain anti-microbial agents and claim to be more effective at removing bacteria than soap bars and conventional liquid soaps. This study attempts to test these claims by comparing the effectiveness of a conventional soap bar, a conventional liquid soap and an anti-microbial liquid soap containing triclosan. In vitro tests demonstrate that the anti-microbial liquid soap is more effective than conventional liquid soaps in reducing the viability of six bacterial species and that this effect is both time and dose dependent. However, when the three soaps were compared for their ability to reduce microbial counts on the hands no differences were observed between the three products. For all three soaps, counts after washing sometimes went up and sometimes down when compared with pre-wash counts. This was the case both when the soaps were used ,normally', that is, with great variation in the time taken, water and soap volumes used and method of washing and after a standardized, rigorous wash recommended in clinical situations. Furthermore, reduction in microbial counts from hands contaminated by handling raw meat was no greater for the anti-microbial than for the conventional liquid soap. [source] Abstracts: The effects of licorice leaf extract on ceramide and hyaluronan synthesisINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 5 2010Akinori Kiso pp.267,273 Both water-holding and permeability barrier function in the stratum corneum (SC) are essential for keeping skin moisture. Intercellular lipids in SC, which are composed mainly of cholesterol, fatty acids, and ceramides, play a crucial role for maintaining the function in SC. The object of our study is to find active ingredients from plant extracts for enhancing the abilities of skin hydration and barrier repair by focusing on the synthesis of ceramides. As a result, we found that licorice leaf extract is a promising ingredient showing not only an increase of mRNA expression levels of serine palmytoyltransferase (SPT) and sphingomyelinase related to ceramide biosynthesis in keratinocytes but also syntheses of ceramides in a 3D skin model and in human skin. Furthermore, licorice leaf extract showed an increase of mRNA expression levels of HMG-CoA reductase (HMGCR) related to cholesterol biosynthesis and an increase of hyaluronan (HA) production in in vitro tests. One of the principles isolated from licorice leaf extract, 6-prenyl-naringenin, was thought to be one of the active components. These results suggested that licorice leaf extract may be a useful ingredient for skin care due to the synthesis of intercellular lipids and HA [source] Study and description of hydrogels and organogels as vehicles for cosmetic active ingredientsINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 4 2010M. E. Morales J. Cosmet. Sci., 60, 627,636 (November/December 2009) Synopsis Cellulite, a clinical syndrome mainly affecting women, involves specific changes in conjunctive dermic and subcutaneous tissue, leading to vascular and hypertrophic alterations in adipose tissues and the consequent alteration of tissue structure. This paper describes the design of hydrogels and pluronic-lecithin organogels elaborated as vehicles of Aloe vera (Aloe vera linné) and Hydrocotyle asiatica (Centella asiatica) for the treatment of cellulite. The objective of this work was to carry out a complete evaluation of the proposed formulae through the study of the organoleptic and rheological properties of the formulae. Our work revealed that, in appearance, hydrogels show better organoleptic characteristics than organogels. On the other hand, from a rheological point of view, both hydrogels and organogels display a plastic behavior. However, the main difference between the two is that the more complex internal structure of the organogel bestows it with more viscosity. Finally, in vitro tests with Franz-type diffusion cells revealed that the release of cosmetic active principle from the tested excipients was appropriate, both in terms of magnitude and velocity. [source] Minireview: does in-vitro testing of oximes help predict their in-vivo action after paraoxon exposure?JOURNAL OF APPLIED TOXICOLOGY, Issue 6 2009D. E. Lorke Abstract K-oximes have recently been developed in the search for efficacious broad-band reactivators of acetylcholinesterase (AChE) inhibited by organophosphorus compounds (OPC). Before clinical use, their toxicity and efficacy need to be assessed, and there is clear demand for simple in vitro tests that can predict in vivo performance. This article summarizes our in vitro data obtained for conventional and experimental oximes in human and rat blood exposed to the OPC paraoxon and correlates them with our in vivo results. The intrinsic AChE inhibitory activity of oximes, as reflected by their in vitro IC50, is strongly correlated with their LD50 (rat): oximes with a high IC50 (K-27, K-48, pralidoxime and obidoxime) also show a high LD50 and are thus relatively non-toxic, whereas oximes K-105, K-108 and K-113 have a low IC50, a low LD50 and are far more toxic. The IC50 is also correlated with the in vivo capacity to protect from paraoxon-induced mortality: oximes with a higher IC50 reduce the relative risk of death more. In contrast, the protective ability as assessed in vitro by the slope of the IC50 shift (tan,), is not correlated with in vivo protection from paraoxon-induced mortality: the best in vivo protectors (K-27 and K-48) show a much lower tan, value (around 2) than K-110 and K-113 (tan, around 10), which hardly reduce the relative risk of death after paraoxon exposure. The partition coefficient logP of the individual oximes is inversely correlated with their IC50 and with their LD50 and is therefore an indicator of toxicity: strongly hydrophilic oximes tend to be less toxic than less hydrophilic ones. These data highlight the good predictive value of in vitro IC50 testing for in vivo toxicity and the limited practical significance of in vitro assessment of protective potency. Copyright © 2009 John Wiley & Sons, Ltd. [source] Rietveld structure and in vitro analysis on the influence of magnesium in biphasic (hydroxyapatite and ,-tricalcium phosphate) mixturesJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 1 2009S. Kannan Abstract The structure of two different Mg-substituted biphasic (HAP and ,-TCP) mixtures along with the biphasic mixtures without substituted Mg2+ was investigated using Rietveld refinement technique. The substituted Mg2+ was found in the ,-TCP phase and its influence on the composition has led to an increase in HAP content of Mg-containing biphasic mixtures when compared with the HAP content detected in pure biphasic mixtures. The refined structural parameters of Ca10(PO4)6(OH)2 and ,-Ca3(PO4)2 confirmed that all the investigated compositions have crystallized in the corresponding hexagonal (space group P63/m) and rhombohedral (space group R3c) structures. The substitution of lower sized magnesium was found preferentially incorporated at the sixfold-coordinated Ca (5) site of ,-TCP, which is due to the strong Ca (5)·O interaction among all the five different Ca sites of ,-Ca3(PO4)2. The in vitro tests using primary culture of osteoblasts showed that all the tested samples are biocompatible and promising materials for in vivo studies. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009 [source] Hydrothermal crystallization of carbonate-containing hydroxyapatite coatings prepared by radiofrequency-magnetron sputtering methodJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 1 2007Satoshi Nakamura Abstract Carbonate-containing hydroxyapatite (HA) films were prepared by low-temperature hydrothermal annealing from carbonate-containing calcium phosphate amorphous coatings on titanium substrates. The biocompatibility of the carbonate-containing HA layers was estimated by in vitro tests using simulated body fluid (SBF). Precursory amorphous coatings were deposited with rf-magnetron sputtering apparatus, using calcium phosphate glass target in Ar/CO2 atmosphere. The carbonate-containing HA coatings were successfully formed by the annealing at above 130°C for 20 h. On the basis of SEM observation, about 2-,m thickness films coated rigidly were durable enough for the hydrothermal treatment. The coating layer was revealed to consist of single phase of PO4, and OH, partially carbonated HA by XRD and IR analyses. Overgrowing of bone-like apatite layers on the carbonate-containing HA surfaces in the SBF implied that the obtained films acquired a sufficient osteoconductivity, while it was still unclear that activity was enhanced, compared to pure HA coatings. The low-temperature hydrothermal annealing method was effective for preparation of rigid HA coatings on titanium as well as modification of their chemical compositions. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2007 [source] Hypersensitivity reactions to penicillins: studies in a group of patients with negative benzylpenicillin G skin testJOURNAL OF CLINICAL PHARMACY & THERAPEUTICS, Issue 3 2009H.-L. Qiao MD PhD Summary Background:, Although skin tests are usually employed to evaluate current penicillin allergy status, a negative result does not exclude hypersensitivity. There is a need for accurate in vitro tests to exclude hypersensitivity. A radioallergosorbent test (RAST) is a potentially good supplementary approach, but there is little information on the suitability of this method to diagnose penicillin hypersensitivity in subjects with a negative skin test to benzylpenicillin. Methods:, A total of 133 patients with a negative skin test to benzylpenicillin G (PG) and all of whom developed allergic reactions to PG were studied. RAST was used to detect eight kinds of specific IgE antibodies to penicillins in serum, which included four kinds of major and minor antigenic determinants to four penicillin drugs. The combination sites for the specific IgE antibodies were studied by RAST inhibition test. Results:, The rate of positive reactions for the specific IgE antibodies was 59·40% (79/133). Of the eight kinds of antigenic determinants, the positive rates for specific IgE against the major and minor determinants were 39·10% (52) and 42·86% (57) respectively. Of the four drugs, positive cases only to PG were 10 (7·5%), were significantly fewer than the cross-reacting positive cases (36) to PG (P < 0·01). In the RAST inhibition studies all drugs exhibited good inhibitory potencies, and in some instances the side-chain of the penicillins could induce specific responses with a variable degree of cross-reactivity among the different penicillins. Conclusion:, Radioallergosorbent test is a good complementary test in persons who are skin-test negative with PG, and the sensitivity of RAST increaes with increasing specificity of IgE antibodies to be detected. 6-APA and the groups, making part of the different side-chains on penicillins, all contributed to the cross-reactivity. [source] A New Stent Design for the Treatment of True Bifurcation Lesions: H-Side Branch StentsJOURNAL OF INTERVENTIONAL CARDIOLOGY, Issue 1 2010MYEONG-KI HONG M.D. Background:There has been much debate for the adequate treatment strategies for true bifurcation lesions. The purpose of this study is to introduce and test a novel stent design for the treatment of true bifurcation lesions. Methods:This side branch stent is composed of three parts: proximal, connecting, and distal parts. The distal part for the side branch vessel has a slope-side stent margin for circumferential coverage of the ostium and one radio-opaque marker for targeting the carina. The proximal part with two radio-opaque markers operates for safe stent delivery and useful guidance for a more precise placement of the distal part on the side branch ostium. Results of the in vitro test in the acrylic resin-made bifurcation phantom model were evaluated with microcomputer tomography. Animal experiments with this new stent platform were also performed in five pigs. Results:In vitro test and microcomputer tomography showed complete coverage of the side branch ostium circumferentially with stent struts, and the absence of stent struts in the main vessel above the side branch ostium level. This side branch stents were successfully deployed in all 5 pigs. The results of animal experiments were also similar to those of in vitro tests. Conclusions:In vivo and vitro tests demonstrated the effective modality of this side branch stent for the treatment of true bifurcation lesions. (J Interven Cardiol 2010;23:54,59) [source] Diagnosis of nonimmediate reactions to ,-lactam antibioticsALLERGY, Issue 11 2004A. Romano Nonimmediate manifestations (i.e. occurring more than 1 h after drug administration), particularly maculopapular and urticarial eruptions, are common during , -lactam treatment. The mechanisms involved in most nonimmediate reactions seem to be heterogeneous and are not yet completely understood. However, clinical and immunohistological studies, as well as analysis of drug-specific T-cell clones obtained from the circulating blood and the skin, suggest that a type-IV (cell-mediated) pathogenic mechanism may be involved in some nonimmediate reactions such as maculopapular or bullous rashes and acute generalized exanthematous pustulosis. In the diagnostic work-up, the patient's history is fundamental; patch testing is useful, together with delayed-reading intradermal testing. The latter appears to be somewhat more sensitive than patch testing, but also less specific. In case of negative allergologic tests, consideration should be given to provocation tests, and the careful administration of the suspect agents. With regard to in vitro tests, the lymphocyte transformation test may contribute to the identification of the responsible drug. Under the aegis of the European Academy of Allergology and Clinical Immunology (EAACI) interest group on drug hypersensitivity and the European Network for Drug Allergy (ENDA), in this review we describe the general guidelines for evaluating subjects with nonimmediate reactions to , -lactams. [source] Management of allergic rhinitis: a common part of practice in primary care clinicsALLERGY, Issue 3 2004D.-Y. Wang Background:, Allergic rhinitis (AR) is an extremely common disease worldwide and one of the top-ten reasons for a visit to primary care clinics. This study aimed to investigate the understanding of current guidelines and concepts of management for AR among general practitioners (GPs) in Singapore. Methods:, A postal questionnaire was designed to survey the dispensing practice and understanding of current guidelines in the management of AR among Singapore GPs. Results:, Two hundred GPs completed the questionnaire. AR was estimated to be 10,40% of total patient visits in 50% of the primary care clinics surveyed. There was no significant difference in diagnosis and management of AR among GPs practicing solo, as a group or in polyclinics. The use of allergy tests (skin or in vitro tests) was <50%. Most physicians understood correctly the efficacy, side-effects, and cost effectiveness of first and newer generation H1-antihistamines and nasal glucocorticosteroids. However, first generation H1-antihistamines and oral/nasal decongestants are commonly used to reduce the cost of medication and to achieve quick relief from nasal obstruction. Conclusion:, Management of allergic rhinitis is common in primary care clinics. Quick symptomatic relief with low-cost medications is a major concern for GPs in the management of AR, as it will affect a patient's compliance and perception of efficacy. However, inappropriate use of decongestants and other nonevidence-based therapies should not be recommended. Implementing the current evidence-based international guidelines for AR needs to be improved. [source] Immediate allergic reactions to cephalosporins and penicillins and their cross-reactivity in childrenPEDIATRIC ALLERGY AND IMMUNOLOGY, Issue 4 2005-Markovi, Marina Atanaskovi Penicillins and cephalosporins are the most important betalactams inducing IgE-mediated reactions. The safety of administering cephalosporins to penicillin-allergic children is a particular problem, because cephalosporin allergenic determinants have not been properly identified. A study was undertaken to evaluate the frequency of anaphylactic reactions to cephalosporins and penicillins and their cross-reactivity in a pediatric population. A prospective survey was conducted in a group of 1170 children with suspected immediate allergic reactions to cephalosporins and/or penicillins, which were examined during a period of 8 yr. In vivo (skin tests and challenges) and in vitro tests (for specific IgE) were performed with standard concentration of penicillins and cephalosporins. When 1170 children with a clinical history of allergy to penicillins and/or cephalosporins were tested in vivo for immediate hypersensitivity to betalactams, 58.3% cases overall were found to be skin or challenge test positive. Among them, 94.4% patients were positive to penicillins and 35.3% to cephalosporins. The frequency of positive reactions in the in vivo testing was in the range from 36.4% to 88.1% for penicillins and from 0.3% to 29.2% for cephalosporins. However, 31.5% of the penicillin allergic children cross-reacted to some cephalosporin. If a child was allergic to a cephalosporin, the frequency of positive reactions to penicillin was 84.2%. The cross-reactivity between cephalosporins and penicillins varied between 0.3% and 23.9%. The cross-reactivity among different generations of cephalosporins varied between 0% and 68.8%, being the highest for first and second-generation cephalosporins and 0% for third generation cephalosporins. The frequency of immediate allergic reactions to cephalosporins is considerably lower compared to penicillins, and the degree of cross-reactivity between cephalosporins and penicillins depends on the generation of cephalosporins, being higher with earlier generation cephalosporins. The cross-reactivity among cephalosporins is lower compared to cross-reactivity between penicillins and cephalosporins. [source] Polymeric Photosensitizer Prodrugs for Photodynamic TherapyPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 4 2007Marino A. Campo ABSTRACT A targeting strategy based on the selective enzyme-mediated activation of polymeric photosensitizer prodrugs (PPP) within pathological tissue has led to the development of agents with the dual ability to detect and treat cancer. Herein, a detailed study of a simple model system for these prodrugs is described. We prepared "first-generation" PPP by directly tethering the photosensitizer (PS) pheophorbide a to poly-(l)-lysine via epsilon amide links and observed that by increasing the number of PS on a polymer chain, energy transfer between PS units improved leading to better quenching efficiency. Fragmentation of the PPP backbone by trypsin digestion gave rise to a pronounced fluorescence increase and to more efficient generation of reactive oxygen species upon light irradiation. In vitro tests using the T-24 bladder carcinoma cell line and ex vivo experiments using mouse intestines illustrated the remarkable and selective ability of these PPP to fluoresce and induce phototoxicity upon enzymatic activation. This work elucidated the basic physicochemical parameters, such as water solubility and quenching/activation behavior, required for the future elaboration of more adaptable "second-generation" PPP, in which the PS is tethered to a proteolytically stable polymer backbone via enzyme-specific peptide linkers. This polymer architecture offers great flexibility to tailor make the PPP to target any pathological tissue known to over-express a specific enzyme. [source] In Vitro Phototoxic Properties of Triamcinolone 16,17-acetonide and Its Main Photoproducts,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 5 2003Giorgia Miolo ABSTRACT The phototoxicity of triamcinolone 16,17-acetonide has been estimated through a panel of,in vitro tests. The main target involved in phototoxicity induced by triamcinolone appeared to be the cell membrane. Oxygen-independent photohemolysis was observed. A photochemical study in water and buffered solutions supported the conclusion that this is related to the action of radicals formed upon UV irradiation (in particular UV-B) by Norrish Type-I fragmentation of the C-20 ketone group. Peroxy radicals were formed in the presence of oxygen and were the active species in that case. Three photoproducts, isolated from the photodegradation of the drug, were submitted to the same toxicity tests. Two of them were proved to possess toxic or phototoxic properties on erythrocytes, primarily induced by UV-B light, and may participate in the photosensitizing activity of triamcinolone 16,17-acetonide. Our in vitro results suggest that the drug can elicit weak photosensitizing properties in vivo. [source] Effects of temperature, sorbitol, alanine and diapause hormone on the embryonic development in Bombyx mori: in vitro tests of old hypothesesPHYSIOLOGICAL ENTOMOLOGY, Issue 4 2005Ken-Ichi Iwata Abstract., An in vitro culture method is described in which embryonic development in Bombyx mori is traced at various temperatures and treatments. The results show that the induction, intensification and termination of diapause are distinct processes. Prediapause embryos, explanted from 40-h-old diapause-destined eggs and cultured in Grace's medium, continue to develop to the appendage-formation stage without arrest, which indicates that the isolated embryos have not entered diapause, whereas the development of embryos from diapausing eggs (15 days after being laid) is significantly slower. The rate of development of embryos dissected from diapause eggs increases during chilling (5 °C) and incubation (at 25 °C) gradually during chilling and dramatically at 25 °C. The in vitro experiments also reveal that sorbitol directly inhibits the development of embryos explanted from diapausing eggs but has no affect on the development of embryos from prediapause eggs. Neither alanine nor diapause hormone prevent isolated embryos from developing. [source] Bioactivity studies on ,-sitosterol and its glucosidePHYTOTHERAPY RESEARCH, Issue 5 2002Irene M. Villaseñor Abstract ,-Sitosterol and ,-sitosteryl-,- D -glucoside were isolated as analgesic constituents from the leaves of Mentha cordifolia Opiz. The acetic acid-induced writhing test showed that ,-sitosterol and ,-sitosteryl-,- D -glucoside decreased the number of squirms induced by acetic acid by 70.0% and 73.0%, respectively, at a dose of 100,mg /,kg mouse. Statistical analysis using the Kruskall Wallis one-way analysis of variance by ranks showed that these isolates approximate the analgesic activity of mefenamic acid at a 0.001 level of significance. The hot plate method confirmed their analgesic activities, as ,-sitosterol and ,-sitosteryl-,- D -glucoside exhibited a 300% and 157% increase in pain tolerance, respectively, while mefenamic acid, a known analgesic, showed a 171% increase. Neither isolate exhibited antiinflammatory activity using the carrageenan-induced mouse paw oedema assay. ,-Sitosterol also exhibited anthelminthic and antimutagenic activities. In vitro tests using live Ascaris suum as test animals showed that the behaviour of worms treated with ,-sitosterol approximated that of the positive controls, Combantrin and Antiox. An in vivo micronucleus test showed that ,-sitosterol inhibited the mutagenicity of tetracycline by 65.3% at a dose of 0.5,mg /kg mouse. At the same dose, it did not exhibit chromosome-breaking activity. Copyright © 2002 John Wiley & Sons, Ltd. [source] | ||||||||||||||||||||||||||||||||||