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Find video protocols related to scientific articles indexed in Pubmed.
Healing Pathways: Longitudinal Effects of Religious Coping and Social support on PTSD Symptoms in African American Sexual Assault Survivors.
J Trauma Dissociation
PUBLISHED: 11-12-2014
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Abstract African American women are at slightly increased risk for sexual assault (Abbey, Jacques-Tiaura, & Parkhill, 2010). However, due to stigma, experiences of racism, and historical oppression, African American women are less likely to seek help from formal agencies when compared to White women (Ullman & Filipas, 2001; Lewis, Resnick, Smith, Best, & Saunders, 2005) and/or women of other ethnic backgrounds (Ahrens, Abeling, Ahmad, & Himman, 2010). Therefore, the provision of culturally appropriate services, such as the inclusion of religion and spiritual coping, may be necessary when working with African American women survivors of sexual assault. The current study, controlling for age and education, explores the impact of religious coping and social support over one year for 252 African American adult female sexual assault survivors recruited from the Chicago metropolitan area. Results from hierarchical linear regression analyses revealed high endorsement of religious coping and social support at Time 1 does not predict a reduction of PTSD symptoms at Time 2. However, high social support at Time 2 does predict lower PTSD at Time 2. Also it is significant to note, survivors with high PTSD at Time 1 and Time 2 endorse greater use of social support and religious coping. Clinical and research implications are explored.
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Statistical Considerations in Setting Product Specifications.
J Biopharm Stat
PUBLISHED: 10-31-2014
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ABSTRACT According to ICH Q6A (1999), a specification is defined as a list of tests, references to analytical procedures, and appropriate acceptance criteria, which are numerical limits, ranges, or other criteria for the tests described. For drug products, specifications usually consist of test methods and acceptance criteria for Assay, Impurities, pH, Dissolution, Moisture, and Microbial Limits, etc., depending on the dosage forms. They are usually proposed by the manufacturers, and subject to the regulatory approval for use. When the acceptance criteria in product specifications cannot be pre-defined based on prior knowledge, the conventional approach is to use data from a limited number of clinical batches during the clinical development phases. Often in time, such acceptance criteria is set as an interval bounded by the sample mean plus and minus two to four standard deviations. This interval may be revised with the accumulated data collected from released batches after drug approval. In this paper, we describe and discuss statistical issues of commonly used approaches in setting or revising specifications (usually tighten the limits), including reference interval, (Min, Max) method, tolerance interval, and confidence limit of percentiles. We also compare their performance in terms of interval width and the intended coverage. Based on our study results and review experiences, we make some recommendations on how to select appropriate statistical methods in setting product specifications to better ensure the product quality.
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Quality Assurance Test of Delivered Dose Uniformity of Multiple-Dose Inhaler and Dry Powder Inhaler Drug Products*
J Biopharm Stat
PUBLISHED: 10-31-2014
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Abstract The delivered dose uniformity is one of the most critical requirements for dry powder inhaler and metered dose inhaler products. In 1999, Food and Drug Administration (FDA) issued a Draft Guidance entitled Nasal Spray and Inhalation Solution, Suspension, and Spray Drug Products-Chemistry, Manufacturing and Controls Documentation and recommended a two-tier acceptance sampling plan that is a modification of the United States Pharmacopeia (USP) sampling plan of dose content uniformity (USP34). This sampling acceptance plan is also applied to Metered Dose Inhaler (MDI) and Dry Powder Inhaler (DPI) Drug Products in general. The FDA Draft Guidance method is shown to have a near-zero probability of acceptance at the second tier. In 2000, under the request of The International Pharmaceutical Aerosol Consortium (IPAC), FDA developed a two-tier sampling acceptance plan based on two one-sided tolerance intervals for small sample. The procedure was presented in the 2005 Advisory Committee Meeting of Pharmaceutical Science and later published in Journal of Biopharmaceutical Statistics (Tsong et al, 2008). This proposed procedure controls the probability of the product delivering below a pre-specified effective dose and the probability of the product delivering over a pre-specified safety dose. In this article, we further propose an extension of the two one-sided tolerance intervals procedure to single tier procedure with any number of canisters.
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Statistical evaluation of several methods for cut point determination of immunogenicity screening assay.
J Biopharm Stat
PUBLISHED: 10-31-2014
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ABSTRACT Cut point of the immunogenicity screening assay is the level of response of the immunogenicity screening assay at or above which a sample is defined to be positive and below which it is defined to be negative. Food and Drug Administration Guidance for Industry on Assay Development for Immunogenicity Testing of Therapeutic recommends the cut point to be an upper 95 percentile of the negative control patients. In this article, we assume that the assay data is a random sample from a normal distribution. The sample normal percentile is a point estimate with variability that decreases with the increase of sample size. Therefore the sample percentile does not assure at least 5% false positive rate with high confidence level (e.g., 90%) when the sample size is not sufficiently enough. With this concern, we propose to use a lower confidence limit for a percentile as the cut point instead. We have conducted an extensive literature review on the estimation of the statistical cut point and compare several selected methods for the immunogenicity screening assay cut point determination in terms of bias, the coverage probability, and false positive rate. The selected methods evaluated for the immunogenicity screening assay cut point determination are sample normal percentile, the exact lower confidence limit of a normal percentile (Charkraborti and Li, 2007), and the approximate lower confidence limit of a normal percentile. It is shown that the actual coverage probability for the lower confidence limit of a normal percentile using approximate normal method is much larger than required confidence level with a small number of assays conducted in practice. We recommend using the exact lower confidence limit of a normal percentile for cut point determination.
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Using Tolerance Intervals for Assessment of Pharmaceutical Quality.
J Biopharm Stat
PUBLISHED: 10-31-2014
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ABSTRACT In quality control of drug products, tolerance intervals are commonly used methods to assure a certain proportion of the products covered within a pre-specified acceptance interval. Depending on the nature of the quality attributes, the corresponding acceptance interval could be one-sided or two-sided. Thus, the tolerance intervals can also be one-sided or two-sided. To better utilize tolerance intervals for quality assurance, we reviewed the computation method and studied their statistical properties in terms of batch acceptance probability in this paper. We also illustrate the application of one-sided and two-sided tolerance, as well as two one-sided test through the examples of dose content uniformity test, delivered dose uniformity test, and dissolution test.
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Rumphellols A and B, new caryophyllene sesquiterpenoids from a Formosan gorgonian coral, Rumphella antipathies.
Int J Mol Sci
PUBLISHED: 09-04-2014
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Two new marine-derived caryophyllene-type sesquiterpenoids, rumphellols A and B (1 and 2), were obtained from the gorgonian coral, Rumphella antipathies, collected off the waters of Taiwan. Although caryophyllene-type sesquiterpenes are rarely found in marine organisms, compounds of this type could be principal components of R. antipathies. The structures of new Compounds 1 and 2 were determined by analysis of their spectroscopic data, including 1D and 2D NMR experiments. Caryophyllene 1 and 2 were evaluated in terms of their anti-inflammatory activity by examining their inhibitory effects on the generation of superoxide anions and the release of elastase by human neutrophils.
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New briarane diterpenoids from Taiwanese soft coral Briareum violacea.
Mar Drugs
PUBLISHED: 08-22-2014
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Ten new briarane diterpenoids, briaviolides A-J (1-10), together with six known briaranes, solenolides A and D, excavatolide A, briaexcavatolide I, 4?-acetoxy-9-deacetystylatulide lactone and 9-deacetylstylatulide lactone, were isolated from the Taiwanese soft coral, Briareum violacea. Their structures were determined on the basis of spectroscopic data ((1)H- and (13)C-NMR, (1)H-(1)H COSY, HSQC, HMBC and NOESY), HR-MS and chemical methods. The absolute configuration of briaviolide A (1) was determined by X-ray crystallographic analysis. Compounds 5, 9 and derivative 11 showed moderate inhibitory activities on superoxide-anion generation and elastase release by human neutrophils in response to N-formyl-methionyl-leucyl-phenylalanine/ Cytochalasin B (fMLP/CB).
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Rumphellaones B and C, new 4,5-seco-caryophyllane sesquiterpenoids from Rumphellan antipathies.
Molecules
PUBLISHED: 08-14-2014
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Two new 4,5-seco-caryophyllane sesquiterpenoids, rumphellaones B (1) and C (2), which were found to possess unprecedented ?-lactone moieties, were obtained from the gorgonian coral Rumphella antipathies. The structures of 1 and 2 were elucidated by spectroscopic methods and compound 2 was found to display modest inhibitory effects on the generation of superoxide anions and the release of elastase by human neutrophils at a concentration of 10 ?g/mL.
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Sample size determination for a three-arm equivalence trial of normally distributed responses.
J Biopharm Stat
PUBLISHED: 08-08-2014
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The equivalence assessment is often conducted through a three-arm clinical trial (namely, test, reference, and placebo) and it usually consists of three tests. The first two tests are to demonstrate the superiority of the test and the reference treatment to the placebo, and they are followed by an equivalence test between the test treatment and the reference treatment. When the response variable is continuous, equivalence is commonly defined in terms of mean difference, mean ratio, or ratio of mean differences, that is, the mean difference of the test and the placebo to the mean difference of the reference and the placebo. These equivalence tests can be performed with both a hypothesis-testing approach and a confidence-interval approach. The advantage of applying the equivalence test by ratio of mean differences is that it can test both superiority of the test treatment over placebo and equivalence between the test and the reference simultaneously through a single hypothesis. In this article, we derive the test statistics and the power function for the ratio of mean differences hypothesis and solve the required sample size for a three-arm clinical trial. Examples of required sample size are given in this article, and are compared with the required sample size by the traditional mean difference equivalence test. After a careful examination, we suggest increasing the power of the ratio of mean differences approach by appropriately adjusting the lower limit of the equivalence interval.
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Seven new sesquiterpenoids from the fruits of Schisandra sphenanthera.
Chem. Biodivers.
PUBLISHED: 07-22-2014
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Fractionation of the EtOH extract from the fruits of Schisandra sphenanthera resulted in the isolation of seven new sesquiterpenoids, 1-7, in addition to the known metabolites 8-23. Among them, schiscupatetralin A (1) possesses an unprecedented structure with a C?C bond between cuparenol and tetralin. The isolated new compounds were evaluated for their anti-HSV-1 and anti-inflammatory activities. The results revealed that compound 4 exhibited anti-HSV-1 activity, while compound 6 showed a significant anti-inflammatory activity.
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Anti-Inflammatory Spirostanol and Furostanol Saponins from Solanum macaonense.
J. Nat. Prod.
PUBLISHED: 07-19-2014
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Eight new spirostanol saponins, macaosides A-H (1-8), and 10 new furostanol saponins, macaosides I-R (9-18), together with six known spirostanol compounds (19-24) were isolated from Solanum macaonense. The structures of the new compounds were determined from their spectroscopic data, and the compounds were tested for in vitro antineutrophilic inflammatory activity. It was found that both immediate inflammation responses including superoxide anion generation and elastase release were significantly inhibited by treatment with compounds 20, 21, and 24 (superoxide anion generation: IC50 7.0, 7.6, 4.0 ?M; elastase release: IC50 3.7, 4.4, 1.0 ?M, respectively). However, compounds 1 and 4 exhibited effects on the inhibition of elastase release only, with IC50 values of 3.2 and 4.2 ?M, respectively, while 19 was active against superoxide anion generation only, with an IC50 value of 6.1 ?M. Accordingly, spirostanols may be promising lead compounds for further neutrophilic inflammatory disease studies.
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Equivalence tests for interchangeability based on two one-sided probabilities.
J Biopharm Stat
PUBLISHED: 07-18-2014
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A test treatment is considered to be interchangeable with its reference treatment if they are equivalent and expected to produce the same clinical result in any given patient. To assess interchangeability, FDA Draft Guidance (1999) and Guidance for Industry (2001, 2003) recommend using individual bioequivalence (IBE) and population bioequivalence (PBE) procedures. Chow (1999) and Chow and Liu (1999) gave a discussion on the limitation of the aggregate criteria of the IBE and PBE proposed therein. They mentioned that it is not clear whether IBE or PBE can imply average bioequivalence. Alternative approaches have been proposed to address the weakness of IBE and PBE. Dong et al. (2014) discuss the tolerance interval method and an approximate test for interchangeability defined by a two-sided probability. These tests may not be able to test for the two one-sided tests (TOST) with asymmetric margins around the true mean difference. In addition, the tests of two-sided probability provide no direction when failing the equivalence in interchangeability. Thus, we reexamine the statistical properties of the two one-sided tolerance interval approaches proposed by Tsong and Shen (2007, 2008). In this project, we extend their approach for parallel arms trials and paired/crossover data without the assumption of equal sample sizes and variances. We also develop the exact power function and assess the type I error rate of our proposed approach. In addition, we study the sample size determination based on the interchangeability testing utilizing the tolerance interval method.
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Equivalence assessment for interchangeability based on two-sided tests.
J Biopharm Stat
PUBLISHED: 07-18-2014
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Interchangeability was originally developed in order to assess drug bioequivalence beyond average bioequivalence. In 2003, the Food and Drug Administration (FDA) published a Guidance documenting the procedures on using in vivo bioequivalence crossover trial to assess interchangeability between test and reference products. In general, this FDA Guidance describes interchangeability in terms of population and individual bioequivalence. The Guidance procedures were criticized for their lack of sampling distribution of the test statistics. As a result, the critical points were generated from simulation studies without adjusting for sample size. Further more, they lack consistency with average bioequivalence required in the 1992 FDA Guidance. Alternative interchangeability or interchangeability procedures were proposed to measure the probability of individual response difference under two treatments within prespecified lower and upper limits. Interchangeability is claimed if this probability is greater than a prespecified threshold. Tse et al. (2006) proposed an approximate distribution of the estimated probability based on the second-order Taylor expansion. For the same interchangeability probability hypothesis, Liu and Chow (1997) and Tsong and Shen (2007) also proposed a tolerance interval-based approach that can be extended to clinical trials with parallel arm design under the normality assumption. In this article, we first generalized the two-sided tolerance interval based interchangeability without equal sample size and variance assumption. We also derived a power function for the proposed method, and performed simulation studies to compare the type I error rate, power, and sample size between the Tse approximated test and the generalized tolerance interval approach for interchangeability assessment.
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Sample size determination for equivalence assessment with multiple endpoints.
J Biopharm Stat
PUBLISHED: 07-18-2014
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Equivalence assessment between a reference and test treatment is often conducted by two one-sided tests (TOST). The corresponding power function and sample size determination can be derived from a joint distribution of the sample mean and sample variance. When an equivalence trial is designed with multiple endpoints, it often involves several sets of two one-sided tests. A naive approach for sample size determination in this case would select the largest sample size required for each endpoint. However, such a method ignores the correlation among endpoints. With the objective to reject all endpoints and when the endpoints are uncorrelated, the power function is the production of all power functions for individual endpoints. With correlated endpoints, the sample size and power should be adjusted for such a correlation. In this article, we propose the exact power function for the equivalence test with multiple endpoints adjusted for correlation under both crossover and parallel designs. We further discuss the differences in sample size for the naive method without and with correlation adjusted methods and illustrate with an in vivo bioequivalence crossover study with area under the curve (AUC) and maximum concentration (Cmax) as the two endpoints.
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The efficacy and safety of cilostazol in ischemic stroke patients with peripheral arterial disease (SPAD): protocol of a randomized, double-blind, placebo-controlled multicenter trial.
Int J Stroke
PUBLISHED: 06-18-2014
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It is not uncommon for patients with ischemic stroke to have peripheral arterial disease (PAD). Patients with polyvascular diseases carry greater burden of atherosclerosis and higher risks of developing vascular events and death. More effective regimens, such as dual antiplatelet agents, may be more effective for controlling progression of atherosclerosis in secondary prevention.
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Excimer Formation in a Confined Space: Photophysics of Ladderphanes with Tetraarylethylene Linkers.
Chemistry
PUBLISHED: 06-04-2014
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Communication between chromophores is vital for both natural and non-natural photophysical processes. Spatial confinements offer unique conditions to scrutinize such interactions. Polynorbornene- and polycyclobutene-based ladderphanes are ideal model compounds in which all tetraarylethylene (TAE) linkers are aligned coherently. The spans for each of the monomeric units in these ladderphanes are 4.5-5.5?Å. Monomers do not exhibit emission, because bond rotation in TAE can quench the excited-state energy. However, polymers emit at 493?nm (?=0.015) with large Stokes shift under ambient conditions and exhibit dual emission at 450 and 493?nm at 150?K. When the temperature is lowered, the emission intensity at 450?nm increases, whereas that at 493?nm decreases. At 100?K, both monomers and polymers emit only at 450?nm. This shorter-wavelength emission arises from the intrinsic emission of TAE chromophore, and the emission at 493?nm could be attributed to the excimer emission in the confined space of ladderphanes. The fast kinetics suggest diffusion-controlled formation of the excimer.
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Complementary Usage of Rhodiola crenulata (L.) in Chronic Obstructive Pulmonary Disease Patients: The Effects on Cytokines and T Cells.
Phytother Res
PUBLISHED: 05-29-2014
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Although chronic obstructive pulmonary disease (COPD) is an inflammatory disease predominantly involving T cells, no study of Rhodiola as an immunomodulator in COPD patients has been reported. In this study, COPD patients took Rhodiola crenulata 500?mg (n?=?38) or placebo (starch/phosphate buffered saline) (n?=?19) daily for 12?weeks and were compared with untreated, age-matched, and sex-matched non-COPD control subjects. Our results showed that serum levels of IL-2, IL-10, and IFN-? in COPD patients before treatment are significantly higher than levels in non-COPD controls (p??0.05). The results suggested that Rhodiola treatment had beneficial antiinflammation effects, lower COPD assessment test score and decreased high-sensitivity C-reactive protein, on COPD patients (p?
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Garcimultiflorone G, a novel benzoylphloroglucinol derivative from Garcinia multiflora with inhibitory activity on neutrophil pro-inflammatory responses.
Chem. Biodivers.
PUBLISHED: 05-16-2014
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A novel benzoylphloroglucinol derivative, garcimultiflorone G (1), was isolated from the fruits of Garcinia multiflora. The structure of 1 was determined through extensive 1D- and 2D-NMR, and MS analyses. Garcimultiflorone G (1) showed inhibitory effects against superoxide anion (O·2(-) generation and elastase release by human neutrophils in response to formyl-L-methionyl-L-leucyl-L-phenylalanine/cytochalasin B (fMLP/CB), with IC50 values of 6.97 ± 1.56 and 11.70 ± 1.58 ?M, respectively.
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Constituents of the roots of Clausena lansium and their potential anti-inflammatory activity.
J. Nat. Prod.
PUBLISHED: 05-05-2014
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Eight new carbazole alkaloids, claulamines C (1), D (2), and E (5) and clausenalines B-F (3, 4, 6-8), four new coumarins, clausemarins A-D (9-12), and 43 known compounds were isolated from the roots of Clausena lansium. The structures of the new compounds were established on the basis of 2D-NMR spectroscopic analysis, and their absolute configurations were established from their ECD spectra. The configuration of wampetin was revised as E using a NOESY experiment. Most of the isolated compounds were evaluated for their potential anti-inflammatory activity. The results showed that compounds 9, 13-18, and 20-22 exhibited strong inhibition of superoxide anion generation with IC50 values ranging from 1.9 to 8.4 ?M, while compounds 18, 19, and 21 inhibited elastase release with IC50 values in the range from 2.0 to 6.9 ?M.
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New cembranoids from the soft coral Sinularia arborea.
Nat Prod Commun
PUBLISHED: 04-03-2014
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From the ethyl acetate extract of the soft coral Sinularia arborea, a new cembrane-type diterpenoid, arbolide C (1), along with (+)-sarcophytol T (2), an enantiomer of the known cembrane, sarcophytol T, were isolated. The structures of compounds 1 and 2 were established by spectroscopic methods and 1 was found to display an inhibitory effect on the release of elastase by human neutrophils.
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Two diterpenoids and a cyclopenta[c]pyridine derivative from roots of Salvia digitaloids.
Int J Mol Sci
PUBLISHED: 04-02-2014
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Two new glucosides, salviadigitoside A (1) and salviatalin A-19-O-?-glucoside (2), belonging to the salviatalin type diterpenoids, and a new cyclopenta[c]pyridine, salviadiginine A (3), were isolated from the roots of Salvia digitaloids. Structures of these compounds were determined on the basis of spectroscopic analysis. In addition, compounds 1-3 were evaluated for their anti-inflammatory activity, but the results showed a weak anti-inflammatory activity.
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In vitro anti-inflammatory effects of diterpenoids and sesquiterpenoids from traditional Chinese medicine Siegesbeckia pubescens.
Bioorg. Med. Chem. Lett.
PUBLISHED: 03-31-2014
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Oxidative stress imposed by reactive oxygen species plays a crucial role in pathophysiology of inflammatory diseases. In the present study, sesquiterpenoids and diterpenoids isolated from Siegesbeckia pubescens, a Chinese traditional medicine used to treat arthritis, were evaluated for inhibition of NO production in activated RAW 264.7 macrophages and FMLP/CB induced O2(·-) generation and elastase release in human neutrophils. In the former assay, sesquiterpenoids were more potent than diterpenoids. The C-4 carbonyl group in the carabrane-type sesquiterpenoid 3 and the C-9 ether linkage in the germacrane sesquiterpene 7 were associated with the enhanced potency. Also, for the active ent-kaurane type diterpenoids, esterification of 17-OH with isobutyric acid and acetylation of 18-OH affected the inhibition of O2(·-) generation and elastase release. This report is the first to describe the inhibitory effects on oxidative stress of secondary metabolites from S. pubescens. Its findings suggest that active terpenoids from the herb could be used as lead anti-inflammatory agents.
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Selective apoptotic cell death effects of oral cancer cells treated with destruxin B.
BMC Complement Altern Med
PUBLISHED: 03-31-2014
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Recent studies have revealed that destruxins (Dtx) have potent cytotoxic activities on individual cancer cells, however, data on oral cancer cells especial human are absent.
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New coumarins and anti-inflammatory constituents from the fruits of Cnidium monnieri.
Int J Mol Sci
PUBLISHED: 03-03-2014
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The fruit of Cnidium monnieri is commercially used as healthcare products for the improvement of impotence and skin diseases. Three new coumarins, 3'-O-methylmurraol (1), rel-(1'S,2'S)-1'-O-methylphlojodicarpin (2), and (1'S,2'S)-1'-O-methylvaginol (3), have been isolated from the fruits of C. monnieri, together with 14 known compounds (4-17). The structures of these new compounds were determined through spectroscopic and MS analyses. Compounds 1, 4-12, and 14-17 exhibited inhibition (IC50 ? 7.31 µg/mL) of superoxide anion generation by human neutrophils in response to formyl-l-methionyl-l-leucyl-l-phenylalanine/cytochalasin B (fMLP/CB). Compounds 7, 9-11, 15, and 17 inhibited fMLP/CB-induced elastase release with IC50 values ?7.83 µg/mL. This investigation reveals that bioactive isolates (especially 6, 7, 14, and 17) could be further developed as potential candidates for the treatment or prevention of various inflammatory diseases.
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Anti-inflammatory triterpenoids from the stems of Microtropis fokienensis.
Molecules
PUBLISHED: 03-03-2014
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Three new ursane- and four new oleanane- type triterpenoids 1-7 were isolated, along with six known compounds 8-13, from the methanolic extract of Microtropis fokienensis. All structures were elucidated by mass and NMR spectroscopic methods. The isolates 4-10 and known compounds 14-17 that were previously isolated from this material were evaluated for anti-inflammatory activity based on effects against superoxide anion generation and elastase release by neutrophils in response to fMLP/CB. 11?,30-Dihydroxy-2,3-seco-olean-12-en-2,3-dioic anhydride (7) was the first triterpene anhydride from the genus of Microtropis to have the ring A expanded to a seven-membered ring; it showed significant anti-inflammatory activity against superoxide anion generation and elastase release. Unexpectedly, 30-hydroxy-2,3-seco-lup-20(29)-ene-2,3-dioic acid (17) showed the best effect against superoxide anion generation and elastase release with IC50 values of 0.06±0.01 and 1.03±0.35 µg/mL, respectively. Compound 17 had a dioic acid function, and compound 7 had an anhydride function modification in ring A; both showed promising activity in the target assays.
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Intracellular glutathione depletion by oridonin leads to apoptosis in hepatic stellate cells.
Molecules
PUBLISHED: 02-17-2014
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Proliferation of hepatic stellate cells (HSCs) plays a key role in the pathogenesis of liver fibrosis. Induction of HSC apoptosis by natural products is considered an effective strategy for treating liver fibrosis. Herein, the apoptotic effects of 7,20-epoxy-ent-kaurane (oridonin), a diterpenoid isolated from Rabdosia rubescens, and its underlying mechanisms were investigated in rat HSC cell line, HSC-T6. We found that oridonin inhibited cell viability of HSC-T6 in a concentration-dependent manner. Oridonin induced a reduction in mitochondrial membrane potential and increases in caspase 3 activation, subG1 phase, and DNA fragmentation. These apoptotic effects of oridonin were completely reversed by thiol antioxidants, N-acetylcysteine (NAC) and glutathione monoethyl ester. Moreover, oridonin increased production of reactive oxygen species (ROS), which was also inhibited by NAC. Significantly, oridonin reduced intracellular glutathione (GSH) level in a concentration- and time-dependent fashion. Additionally, oridonin induced phosphorylations of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK). NAC prevented the activation of MAPKs in oridonin-induced cells. However, selective inhibitors of MAPKs failed to alter oridonin-induced cell death. In summary, these results demonstrate that induction of apoptosis in HSC-T6 by oridonin is associated with a decrease in cellular GSH level and increase in ROS production.
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Eunicellin-based diterpenoids, hirsutalins N-R, from the formosan soft coral Cladiella hirsuta.
Mar Drugs
PUBLISHED: 02-13-2014
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New eunicellin-type hirsutalins N-R (1-5), along with two known eunicellins, (6 and 7) were isolated from the soft coral Cladiella hirsuta. The structures of the metabolites were determined by extensive spectroscopic analysis. Cytotoxic activity of compounds 1-7 against the proliferation of a limited panel of cancer cell lines was measured. The in vitro anti-inflammatory activity of compounds 1-7 was evaluated by measuring their ability in suppressing superoxide anion generation and elastase release in fMLP/CB-induced human neutrophils.
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Tortuosenes A and B, new diterpenoid metabolites from the Formosan soft coral Sarcophyton tortuosum.
Org. Lett.
PUBLISHED: 02-13-2014
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Tortuosenes A and B (1 and 2), possessing new diterpenoid molecular structures, were isolated from the Formosan soft coral Sarcophyton tortuosum along with a known cembrane, emblide (3). The structures of 1 and 2 were established by extensive spectroscopic analysis, and the absolute configuration of 1 was determined by TDDFT ECD calculations. Tortuosene A was found to display significant inhibitory effects on the generation of the superoxide anion in N-formyl-methionyl-leucyl-phenylalanine/cytochalasin B (fMLP/CB)-induced neutrophils.
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A novel immunomodulatory effect of ugonin U in human neutrophils via stimulation of phospholipase C.
Free Radic. Biol. Med.
PUBLISHED: 01-26-2014
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Neutrophils have a crucial role in the immune system and are the first line of defense against pathogenic invaders. Neutrophil activation is required for their defensive function and can be induced by diverse stimuli, through either binding to cell surface receptors or direct intracellular target molecule stimulation. In this study, we found that 4?a,5?,6?,7?,8?,8?a-hexahydro-5,3',4'-trihydroxy-5?,5?,8?a-trimethyl-4H-chromeno [2?,3?:7,6]flavone (ugonin U), a flavonoid isolated from Helminthostachys zeylanica (L) Hook, significantly induced superoxide production and release in a time- and concentration-dependent manner. A series of experiments was performed to dissect the mechanism of ugonin U-induced respiratory burst in human neutrophils. Our results demonstrated that ugonin U induced a slow increase in intracellular Ca(2+), which was necessary for ugonin U-stimulated superoxide release. Use of a formyl peptide receptor (FPR) blocker, G protein inhibitor, and protein tyrosine kinase (PTK) inhibitor proved that FPR, G proteins, and PTKs were not associated with ugonin U-induced respiratory burst. Additionally, immunoblotting results revealed that ugonin U did not affect the phosphorylation of mitogen-activated protein kinases and protein tyrosine. Nevertheless, a phospholipase C (PLC) inhibitor and an inositol triphosphate (IP3) receptor antagonist considerably suppressed ugonin U-stimulated Ca(2+) mobilization and subsequent superoxide release. Ugonin U also induced an increase in intracellular IP3 formation, which could be blocked using a PLC inhibitor. In conclusion, our study reveals that ugonin U represents the first identified natural flavonoid compound to directly stimulate PLC. Moreover, ugonin U induces respiratory burst via the PLC/IP3/Ca(2+) pathway in human neutrophils.
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Krempfielins N-P, New anti-inflammatory eunicellins from a Taiwanese soft coral Cladiella krempfi.
Mar Drugs
PUBLISHED: 01-22-2014
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Three new eunicellin-type diterpenoids, krempfielins N-P (1-3), were isolated from a Taiwanese soft coral Cladiella krempfi. The structures of the new metabolites were elucidated by extensive spectroscopic analysis and by comparison with spectroscopic data of related known compounds. Compound 3 exhibited activity to inhibit superoxide anion generation. Both 1 and 3, in particular 1, were shown to display significant anti-inflammatory activity by inhibiting the elastase release in FMLP/CB-induced human neutrophils.
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Trinorditerpenes from the roots of Flueggea virosa.
Bioorg. Med. Chem. Lett.
PUBLISHED: 01-07-2014
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Two trinorditerpenes, flueggrenes A and B (1 and 2), have been isolated from the roots of Flueggea virosa. Their structures were established by extensive analyses of spectroscopic data. The isolates were evaluated for anti-HCV activity, as well as the inhibition of superoxide anion generation and elastase release in response to FMLP/cytochalasin B.
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Discovery of novel diterpenoids from Sinularia arborea.
Mar Drugs
PUBLISHED: 01-03-2014
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Two novel diterpenoids, sinularbols A (1) and B (2), which were found to possess a new carbon skeleton were isolated from the soft coral Sinularia arborea. The structures of compounds 1 and 2 were elucidated by spectroscopic methods and 2 displayed a moderately inhibitory effect on the generation of superoxide anion by human neutrophils.
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Ursolic Acid inhibits superoxide production in activated neutrophils and attenuates trauma-hemorrhage shock-induced organ injury in rats.
PLoS ONE
PUBLISHED: 01-01-2014
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Neutrophil activation is associated with the development of organ injury after trauma-hemorrhagic shock. In the present study, ursolic acid inhibited the superoxide anion generation and elastase release in human neutrophils. Administration of ursolic acid attenuated trauma-hemorrhagic shock-induced hepatic and lung injuries in rats. In addition, administration of ursolic acid attenuated the hepatic malondialdehyde levels and reduced the plasma aspartate aminotransferase and alanine aminotransferase levels after trauma-hemorrhagic shock. In conclusion, ursolic acid, a bioactive natural compound, inhibits superoxide anion generation and elastase release in human neutrophils and ameliorates trauma-hemorrhagic shock-induced organ injury in rats.
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Natural clovanes from the gorgonian coral Rumphella antipathies.
Nat Prod Commun
PUBLISHED: 10-02-2013
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Three natural clovane-related sesquiterpenoids, 2beta-acetoxyclovan-9alpha-ol (1), 9alpha-acetoxyclovan-2beta-ol (2) and clovan-2beta,9beta-diol (3), were isolated from the gorgonian coral Rumphella antipathies. The structures of clovanes 1-3 were elucidated by spectroscopic methods and by comparison of the spectral data with those of known clovane analogues. This is the first time that clovanes 1-3 have been isolated from a natural source. Clovanes 1 and 2 displayed inhibitory effects on the generation of superoxide anions and the release of elastase by human neutrophils.
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Triterpenoids and steroids from Ganoderma mastoporum and their inhibitory effects on superoxide anion generation and elastase release.
Molecules
PUBLISHED: 09-27-2013
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The methanol extracts of the fruiting bodies of Ganoderma mastoporum collected in Vietnam was purified to afford eight compounds, including three triterpenoids and five steroids. The purified compounds were examined for their inhibitory effects against superoxide anion generation and elastase release. Among the tested compounds, ergosta-4,6,8(14),22-tetraen-3-one (3) exhibited the most significant inhibition towards superoxide anion generation and elastase release with IC50 values of 2.30 ± 0.38 and 1.94 ± 0.50 µg/mL, respectively.
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Eunicellin-based diterpenoids from the Formosan soft coral Klyxum molle with inhibitory activity on superoxide generation and elastase release by neutrophils.
J. Nat. Prod.
PUBLISHED: 09-10-2013
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Eleven new eunicellin-based diterpenoids possessing a cladiellane skeleton with a C-2, C-9 ether bridge, klymollins I-S (1-11), have been isolated from the EtOAc extract of the soft coral Klyxum molle from Taiwan waters. The structures of compounds 1-11 were elucidated by extensive spectroscopic analysis, including 2D NMR spectroscopy (COSY, HSQC, HMBC, and NOESY). Compound 5 exhibited cytotoxicity toward several cancer cell lines. Compound 5 is the first eunicellin-based metabolite bearing a phenyl group and displays significant inhibition of both superoxide anion generation and elastase release in N-formyl-methionyl-leucyl-phenylalanine/cytochalasin B (fMLP/CB)-induced human neutrophils.
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New benzo[c]phenanthridine and benzenoid derivatives, and other constituents from Zanthoxylum ailanthoides: Effects on neutrophil pro-inflammatory responses.
Int J Mol Sci
PUBLISHED: 08-06-2013
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A new benzo[c]phenanthridine, oxynorchelerythrine (1), and two new benzenoid derivatives, methyl 4-(2-hydroxy-4-methoxy-3-methyl-4-oxobutoxy)benzoate (2) and (E)-methyl 4-(4-((Z)-3-methoxy-3-oxoprop-1-enyl)phenoxy)-2-methylbut-2-enoate (3), have been isolated from the twigs of Zanthoxylum ailanthoides, together with 11 known compounds (4-14). The structures of these new compounds were determined through spectroscopic and MS analyses. Among the isolated compounds, decarine (4), (-)-syringaresinol (6), (+)-episesamin (8), glaberide I (9), (-)-dihydrocubebin (10), and xanthyletin (11) exhibited potent inhibition (IC50 values ? 4.79 µg/mL) of superoxide anion generation by human nutrophils in response to N-formyl-L-methionyl-L-leucyl-L-phenylalanine/cytochalasin B (fMLP/CB). Compounds 4, 8, and 11 also inhibited fMLP/CB-induced elastase release with IC50 values ? 5.48 µg/mL.
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Reevesioside F induces potent and efficient anti-proliferative and apoptotic activities through Na?/K?-ATPase ?3 subunit-involved mitochondrial stress and amplification of caspase cascades.
Biochem. Pharmacol.
PUBLISHED: 08-05-2013
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Reevesioside F, isolated from Reevesia formosana, induced anti-proliferative activity that was highly correlated with the expression of Na?/K?-ATPase ?? subunit in several cell lines, including human leukemia HL-60 and Jurkat cells, and some other cell lines. Knockdown of ?? subunit significantly inhibited cell apoptosis suggesting a crucial role of the ?? subunit. Reevesioside F induced a rapid down-regulation of survivin protein, followed by release of cytochrome c from mitochondria and loss of mitochondrial membrane potential (??m). Further examination demonstrated the mitochondrial damage in leukemic cells through Mcl-1 down-regulation, Noxa up-regulation and an increase of the formation of truncated Bid, tBim and a 23-kDa cleaved Bcl-2 fragment. Furthermore, reevesioside F induced an increase of mitochondria-associated acetyl ?-tubulin that may also contribute to apoptosis. The caspase cascade was profoundly activated by reevesioside F. Notably, the specific caspase-3 inhibitor z-DEVD-fmk significantly blunted reevesioside F-induced loss of ??m and apoptosis, suggesting that caspase-3 activation may further amplify mitochondrial damage and apoptotic signaling cascade. In spite of being a cardiac glycoside, reevesioside F did not increase the intracellular Ca²? levels. Moreover, CGP-37157 which blocked Na?/Ca²? exchanger on plasma membrane and mitochondria did not modify reevesioside F-mediated effect. In summary, the data suggest that reevesioside F induces apoptosis through the down-regulation of survivin and Mcl-1, and the formation of pro-apoptotic fragments from Bcl-2 family members. The loss of ??m and mitochondrial damage are responsible for the activation of caspases. Moreover, the amplification of caspase-3-mediated signaling pathway contributes largely to the execution of apoptosis in leukemic cells.
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Cationic surfactants in the form of nanoparticles and micelles elicit different human neutrophil responses: A toxicological study.
Colloids Surf B Biointerfaces
PUBLISHED: 07-07-2013
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Cationic surfactants are an ingredient commonly incorporated into nanoparticles for clinical practicability; however, the toxicity of cationic surfactants in nanoparticles is not fully elucidated. We aimed to evaluate the inflammatory responses of cationic nanobubbles and micelles in human neutrophils. Soyaethyl morpholinium ethosulfate (SME) and hexadecyltrimethyl-ammonium bromide (CTAB) are the two cationic surfactants employed in this study. The zeta potential of CTAB nanobubbles was 80mV, which was the highest among all formulations. Nanobubbles, without cationic surfactants, showed no cytotoxic effects on neutrophils in terms of inflammatory responses. Cationic nanobubbles caused a concentration-dependent cytotoxicity of degranulation (elastase release) and membrane damage (release of lactate dehydrogenase, LDH). Among all nanoparticles and micelles, CTAB-containing nanosystems showed the greatest inflammatory responses. A CTAB nanobubble diluent (1/150) increased the LDH release 80-fold. Propidium iodide staining and scanning electron microscopy (SEM) verified cell death and morphological change of neutrophils treated by CTAB nanobubbles. SME, in a micelle form, strengthened the inflammatory response more than SME-loaded nanobubbles. Membrane interaction and subsequent Ca(2+) influx were the mechanisms that triggered inflammation. The information obtained from this work is beneficial in designing nanoparticulate formulations for balancing clinical activity and toxicity.
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Suberoylanilide hydroxamic acid, a histone deacetylase inhibitor, induces the production of anti-inflammatory cyclodepsipeptides from Beauveria felina.
J. Nat. Prod.
PUBLISHED: 07-03-2013
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The addition of the histone deacetylase inhibitor suberoylanilide hydroxamic acid to a culture of the filamentous fungus Beauveria felina significantly changed its secondary metabolite profile and led to the isolation of eight compounds, including three new cyclodepsipeptides, desmethylisaridin E (1), desmethylisaridin C2 (2), and isaridin F (3), along with five known cyclodepsipeptide compounds. Isaridin F (3) possesses a cyclodepsipeptide ring with N-methylbutyric acid, which is rare in natural peptides. Absolute configurations of the new cyclodepsipeptides were achieved by Marfeys method. The anti-inflammatory activity of the isolated compounds was investigated through evaluating their effect on superoxide anion production and elastase release by FMLP-induced human neutrophils. Among the tested compounds, desmethylisaridin E (1) inhibited superoxide anion production and desmethylisaridin C2 (2) inhibited elastase release, with IC50 values of 10.00 ± 0.80 and 10.01 ± 0.46 ?M, respectively.
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Two anti-inflammatory steroidal saponins from Dracaena angustifolia Roxb.
Molecules
PUBLISHED: 06-18-2013
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Two new steroidal saponins, named drangustosides A-B (1-2), together with eight known compounds were isolated and characterized from the MeOH extract of Dracaena angustifolia Roxb. The structures of compounds were assigned based on 1D and 2D NMR spectroscopic analyses, including HMQC, HMBC, and NOESY. Compounds 1 and 2 showed anti-inflammatory activity by superoxide generation and elastase release by human neutrophils in response to fMLP/CB.
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Krempfielins J-M, new eunicellin-based diterpenoids from the soft coral Cladiella krempfi.
Mar Drugs
PUBLISHED: 06-17-2013
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New four eunicellin-based diterpenoids, krempfielins J-M (1-4) were isolated from the organic extract of a Taiwanese soft coral Cladiella krempfi. The structures of the new metabolites were elucidated on the basis of extensive spectroscopic analysis. The structure of compound 2 is rare due to the presence of the highly oxygenated pattern. Anti-inflammatory activity of 1-6 to inhibit the superoxide anion generation and elastase release in FMLP/CB-induced human neutrophils was also evaluated, and 2 and 4 were shown to possess the ability to inhibit the elastase release.
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Propofol inhibits superoxide production, elastase release, and chemotaxis in formyl peptide-activated human neutrophils by blocking formyl peptide receptor 1.
J. Immunol.
PUBLISHED: 05-13-2013
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Neutrophils play a critical role in acute and chronic inflammatory processes, including myocardial ischemia/reperfusion injury, sepsis, and adult respiratory distress syndrome. Binding of formyl peptide receptor 1 (FPR1) by N-formyl peptides can activate neutrophils and may represent a new therapeutic target in either sterile or septic inflammation. Propofol, a widely used i.v. anesthetic, has been shown to modulate immunoinflammatory responses. However, the mechanism of propofol remains to be established. In this study, we showed that propofol significantly reduced superoxide generation, elastase release, and chemotaxis in human neutrophils activated by fMLF. Propofol did not alter superoxide generation or elastase release in a cell-free system. Neither inhibitors of ?-aminobutyric acid receptors nor an inhibitor of protein kinase A reversed the inhibitory effects of propofol. In addition, propofol showed less inhibitory effects in non-FPR1-induced cell responses. The signaling pathways downstream from FPR1, involving calcium, AKT, and ERK1/2, were also competitively inhibited by propofol. These results show that propofol selectively and competitively inhibits the FPR1-induced human neutrophil activation. Consistent with the hypothesis, propofol inhibited the binding of N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein, a fluorescent analog of fMLF, to FPR1 in human neutrophils, differentiated THP-1 cells, and FPR1-transfected human embryonic kidney-293 cells. To our knowledge, our results identify, for the first time, a novel anti-inflammatory mechanism of propofol by competitively blocking FPR1 in human neutrophils. Considering the importance of N-formyl peptides in inflammatory processes, our data indicate that propofol may have therapeutic potential to attenuate neutrophil-mediated inflammatory diseases by blocking FPR1.
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Design and synthesis of tryptophan containing dipeptide derivatives as formyl peptide receptor 1 antagonist.
Org. Biomol. Chem.
PUBLISHED: 05-01-2013
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Our previous studies identified an Fmoc-(S,R)-tryptophan-containing dipeptide derivative, 1, which selectively inhibited neutrophil elastase release induced by formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) in human neutrophils. In an attempt to improve pharmacological activity, a series of tryptophan-containing dipeptides were synthesized and their pharmacological activities were investigated in human neutrophils. Of these, five compounds 3, 6, 19a, 24a, and 24b exhibited potent and dual inhibitory effects on FMLP-induced superoxide anion (O2?(-)) generation and neutrophil elastase release in neutrophils with IC50 values of 0.23/0.60, 1.88/2.47, 1.87/3.60, 0.12/0.37, and 1.32/1.03 ?M, respectively. Further studies indicated that inhibition of superoxide production in human neutrophils by these dipeptides was associated with the selective inhibition of formyl peptide receptor 1 (FPR1). Furthermore, the results of structure-activity relationship studies concluded that the fragment N-benzoyl-Trp-Phe-OMe (3) was most suitable as a core structure for interaction with FPR1, and may be approved as a lead for the development of new drugs in the treatment of neutrophilic inflammatory diseases. As some of the synthesized compounds exhibited separable conformational isomers, and showed diverse bioactivities, the conformation analysis of these compounds is also discussed herein.
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Bioactive secondary metabolites of a marine Bacillus sp. inhibit superoxide generation and elastase release in human neutrophils by blocking formyl peptide receptor 1.
Molecules
PUBLISHED: 04-25-2013
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It is well known that overwhelming neutrophil activation is closely related to acute and chronic inflammatory injuries. Formyl peptide receptor 1 (FPR1) plays an important role in activation of neutrophils and may represent a potent therapeutic target in inflammatory diseases. In the present study, we demonstrated that IA-LBI07-1 (IA), an extract of bioactive secondary metabolites from a marine Bacillus sp., has anti-inflammatory effects in human neutrophils. IA significantly inhibited superoxide generation and elastase release in formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP)-activated neutrophils, but failed to suppress the cell responses activated by non-FPR1 agonists. IA did not alter superoxide production and elastase activity in cell-free systems. IA also attenuated the downstream signaling from FPR1, such as the Ca2+, MAP kinases and AKT pathways. In addition, IA inhibited the binding of N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein, a fluorescent analogue of FMLP, to FPR1 in human neutrophils and FPR1-transfected HEK293 cells. Taken together, these results show that the anti-inflammatory effects of IA in human neutrophils are through the inhibition of FPR1. Also, our data suggest that IA may have therapeutic potential to decrease tissue damage induced by human neutrophils.
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New Labdane-Type Diterpenoids and Anti-Inflammatory Constituents from Hedychium coronarium.
Int J Mol Sci
PUBLISHED: 04-16-2013
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Four new labdane-type diterpenoids: hedychicoronarin (1), peroxycoronarin D (2), 7?-hydroxycalcaratarin A (3), and (E)-7?-hydroxy-6-oxo-labda-8(17),12-diene-15,16-dial (4), have been isolated from the rhizomes of Hedychium coronarium, together with 13 known compounds (5-17). The structures of these new compounds were determined through spectroscopic and MS analyses. Compounds 3, 5, 6, and 10 exhibited inhibition (IC50 values ?4.52 ?g/mL) of superoxide anion generation by human neutrophils in response to formyl-L-methionyl-L-leucyl-L-phenylalanine/cytochalasin B (fMLP/CB). Compounds 3-6, 10, and 11 inhibited fMLP/CB-induced elastase release with IC50 values ?6.17 ?g/mL.
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Four new briarane diterpenoids from Taiwanese gorgonian Junceella fragilis.
Mar Drugs
PUBLISHED: 04-15-2013
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Four new 8-hydroxybriarane diterpenoids, frajunolides P-S (1-4), together with umbraculolide A, juncenolide C, junceellonoid A and juncin R, were isolated from the acetone extract of the gorgonian Junceella fragilis, collected from the southeast coast of Taiwan. Compound 1 contains an unusual pivaloyloxy group at C-2, while 3 is a rare compound having a chlorine atom on the olefinic carbon (C-6). The structures of the isolated compounds were established by extensive spectroscopic analysis, including 1D- and 2D-NMR, as well as HRMS data. Compound 1 was further confirmed by X-ray crystallographic analysis. In the anti-inflammatory test, compounds 1 and 2 exhibited moderate inhibition on superoxide anion generation and elastase release by human neutrophils in response to formylmethionylleucyl-phenylalanine/dihydrocytochalasin B (fMLP/CB).
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A home-based training program improves caregivers skills and dementia patients aggressive behaviors: a randomized controlled trial.
Am J Geriatr Psychiatry
PUBLISHED: 04-06-2013
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To investigate the effects of an individualized, home-based caregiver-training program for caregivers of elderly patients with dementia and behavioral problems.
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Anti-neutrophilic inflammatory steroidal glycosides from Solanum torvum.
Phytochemistry
PUBLISHED: 03-29-2013
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Torvpregnanosides A and B, two pregnane glycosides, and torvoside Q, a 23-keto-spirostanol glycoside, along with twelve known steroidal saponins were isolated from aerial parts of Solanum torvum. Of the latter, four of the 23-hydroxy-spirostanol glycosides, and, a yamogenin glycoside, were in this plant discovered. All structures were identified from spectroscopic data, and all the compounds were tested for in vitro anti-neutrophilic inflammatory activity. Two compounds showed selective inhibition against elastase release and superoxide anion generation, respectively, by human neutrophils with IC50 values of 5.66 and 3.59 ?M, while two others inhibited both inflammatory mediators with IC50 values of 0.66-3.49 ?M. Structure-activity relationships are discussed.
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Phyto-SERM constitutes from Flemingia macrophylla.
Int J Mol Sci
PUBLISHED: 03-15-2013
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The methanolic extract of Flemingia macrophylla roots exhibited significant estrogenic activity in the transgenic plant assay system which was comparable to the activity of soybean extract. Utilizing estrogenic activity-guided fractionation, one new compound, fleminigin, together with 23 known compounds were isolated from F. macrophylla roots methanolic extract. The structure of the new compound was identified based on intensive spectroscopic analysis and the full spectral data for one of the isolated compounds, flemichin E, was introduced for the first time in the current investigation. The estrogenic and anti-estrogenic activities of the isolated compounds were evaluated revealing that the isolated isoflavonoids may act as partial estrogen agonists, as well as antagonists. Additionally, the anti-inflammatory and the cytotoxic activities of the isolated compounds were studied. These results suggested the potential applications of F. macrophylla extract and its isolated compounds as selective estrogen receptor modulators (SERMs).
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An epigenetic modifier enhances the production of anti-diabetic and anti-inflammatory sesquiterpenoids from Aspergillus sydowii.
Bioorg. Med. Chem.
PUBLISHED: 03-14-2013
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The addition of a DNA methyltransferase inhibitor, 5-azacytidine, to Aspergillus sydowii fungus culture broth changed its secondary metabolites profile. Analysis of the culture broth extract led to the isolation of three new bisabolane-type sesquiterpenoids: (7S)-(+)-7-O-methylsydonol (1), (7S,11S)-(+)-12-hydroxysydonic acid (2) and 7-deoxy-7,14-didehydrosydonol (3), along with eight known compounds. The isolated compounds were evaluated for their anti-diabetic and anti-inflammatory activities. Among the isolates, (S)-(+)-sydonol (4) did not only potentiate insulin-stimulated glucose consumption but also prevented lipid accumulation in 3T3-L1 adipocytes. Additionally, (S)-(+)-sydonol (4) exhibited significant anti-inflammatory activity through inhibiting superoxide anion generation and elastase release by fMLP/CB-induced human neutrophils. This is the first report on isolating a secondary metabolite with anti-diabetic and anti-inflammatory activities from microorganisms.
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Non-invasive synergistic treatment of brain tumors by targeted chemotherapeutic delivery and amplified focused ultrasound-hyperthermia using magnetic nanographene oxide.
Adv. Mater. Weinheim
PUBLISHED: 03-07-2013
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The combination of chemo-thermal therapy is the best strategy to ablate tumors, but how to heat deep tumor tissues effectively without side-damage is a challenge. Here, a systemically delivered nanocarrier is designed with multiple advantages, including superior heat absorption, highly efficient hyperthermia, high drug capacity, specific targeting ability, and molecular imaging, to achieve both high antitumor efficacy and effective amplification of hyperthermia with minimal side effects.
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Flexibilins A-C, new cembrane-type diterpenoids from the Formosan soft coral, Sinularia flexibilis.
Mar Drugs
PUBLISHED: 03-05-2013
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Three new cembrane-type diterpenoids, flexibilins A-C (1-3), along with a known cembrane, (-)-sandensolide (4), were isolated from the soft coral, Sinularia flexibilis. The structures of cembranes 1-4 were elucidated by spectroscopic methods. The structure of 4, including its absolute stereochemistry, was further confirmed by single-crystal X-ray diffraction analysis. Cembrane 2 displayed a moderate inhibitory effect on the release of elastase by human neutrophils.
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Hepatitis B virus X protein disrupts stress fiber formation and triggers apoptosis.
Virus Res.
PUBLISHED: 02-19-2013
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Cytoskeletal proteins are key participants in the cellular progression to apoptosis. In a previous study we injected nude mice with CCL13-HBx cells and identified in contrast to non-HBx transfected cells a differentially phosphorylated myosin light chain (p-MLC) by two-dimensional PAGE and mass spectrometry of the tumor material. To investigate the role of HBx in myosin light chain kinase (MLCK) signaling pathways, we analyzed the key molecules, p-MLC and MLCK, by western blotting. Immunofluorescence staining analysis showed that HBx disrupted stress fiber formation and that focal adhesion kinase (FAK) and integrin-linked kinase (ILK) were regulated by HBx-mediated phosphatase and tensin homolog (PTEN). We also used pharmacological inhibitors to explore the correlation between cytoskeletal rearrangements and HBx-mediated cell apoptosis via an MLCK and a PTEN-dependent pathway. The results showed that both ML9 and bvp restored the effects caused by HBx induction. Our findings suggest that HBx disrupts stress fiber formation and triggers apoptosis via an MLCK and a PTEN-dependent pathway.
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A new hydroxychavicol dimer from the roots of Piper betle.
Molecules
PUBLISHED: 02-08-2013
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A new hydroxychavicol dimer, 2-(g-hydroxychavicol)-hydroxychavicol (1), was isolated from the roots of Piper betle Linn. along with five known compounds, hydroxychavicol (2), aristololactam A II (3), aristololactam B II (4), piperolactam A (5) and cepharadione A (6). The structures of these isolated compounds were elucidated by spectroscopic methods. Compounds 1 and 2 exhibited inhibitory effects on the generation of superoxide anion and the release of elastase by human neutrophils.
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Maximizing dermal targeting and minimizing transdermal penetration by magnolol/honokiol methoxylation.
Int J Pharm
PUBLISHED: 01-24-2013
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Magnolol and honokiol, predominant active compounds in the family Magnoliaceae, are known to exhibit strong anti-inflammatory activities against dermal disorders. We attempted to modify the structures of magnolol and honokiol by methoxylation to optimize the skin delivery ability. Absorption of these permeants into and through the skin was performed at both an infinite dose and saturated solubility. Superoxide anion and elastase released from human neutrophils were the biomarkers used to examine anti-inflammatory potencies of these permeants. The safety of the permeants was evaluated by keratinocyte viability and in vivo bioengineering techniques. Topical magnolol and honokiol at an infinite dose (7.5 mM) showed skin accumulations of 0.22 and 0.16 nmol/mg, respectively. Methoxylation significantly enhanced their skin absorption. Deposition amounts of dimethylmagnolol and dimethylhonokiol were respectively 15- and 7-fold greater than those of magnolol and honokiol. Contrary to the skin accumulation results, the transdermal penetration across skin decreased following methoxylation. No transdermal delivery occurred for dimethylhonokiol. Skin uptake of 4-O-methylhonokiol was 2-fold higher than that of 2-O-methylhonokiol, although they are isomers. Methoxylated permeants demonstrated selective absorption into follicles, which showed 3-5-fold higher follicular amounts compared to magnolol and honokiol. The relative order of anti-inflammatory activities was honokiol>2-O-methylmagnolol>dimethylhonokiol>magnolol. The other compounds exhibited negligible or negative responses in activated neutrophils. Magnolol and honokiol induced slight but significant keratinocyte cytotoxicity and stratum corneum disruption. Daily administration of methoxylated permeants, especially dimethylhonokiol, produced no skin irritation for up to 7 days. Methoxylated magnolol and honokiol can be efficient and safe candidates for treating inflammatory skin disorders.
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Anti-inflammatory diterpenoids from Croton tonkinensis.
J. Nat. Prod.
PUBLISHED: 01-24-2013
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Phytochemical investigation of the methanolic extract of Croton tonkinensis afforded two known kauranes (1, 2), eight new ent-kauranes (3-10), and 16 known ent-kaurane-type diterpenoids (12-27). In addition, 30 known compounds were identified by comparison of their physical and spectroscopic data with reported data. Among the isolated compounds, ent-18-acetoxykaur-16-en-15-one (20) displayed the most significant inhibition of superoxide anion generation and elastase release.
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1,5-Diphenylpent-3-en-1-ynes and methyl naphthalene carboxylates from Lawsonia inermis and their anti-inflammatory activity.
Phytochemistry
PUBLISHED: 01-23-2013
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Lawsonia inermis (Lythraceae) known as henna is one of the most popular and ancient plants used in cosmetics and hair dying. It is cultivated for its leaves but other parts such as seeds, flowers, stem bark and roots are also used in traditional medicine for millennia. Henna tattoo paste also proved to be beneficial for wound healing and in several skin diseases suggesting potent anti-inflammatory activity. To evaluate henna anti-inflammatory activity, 31 compounds, including three 1,5-diphenylpent-3-en-1-yne derivatives, lawsochylin A-C and three methyl naphthalene carboxylates, lawsonaphthoate A-C, were isolated from the stems and leaves of henna utilizing a bioassay-guided fractionation. The structures of the compounds were elucidated by spectroscopic data. Two compounds, lawsochylin A and lawsonaphthoate A showed potent anti-inflammatory activity by inhibition of superoxide anion generation (IC(50)=1.80 and 1.90 ?g/ml) and elastase release (IC(50)=1.58 and 3.17 ?g/ml) of human neutrophils in response to fMLP or cytochalasin B. Moreover, the known compounds, luteolin, apigenin, 4S-4-hydroxy-?-tetralone, and 2-butoxysuccinic acid, also showed potent inhibition of superoxide anion generation (IC(50)=0.75-1.78 ?g/ml) and elastase release (IC(50)=1.62-3.61 ?g/ml).
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Isolation and synthesis of melodamide A, a new anti-inflammatory phenolic amide from the leaves of Melodorum fruticosum.
Planta Med.
PUBLISHED: 01-23-2013
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Together with twelve known compounds (2-13), melodamide A (1), a new phenolic amide possessing p-quinol moiety, was purified and characterized from the methanolic extracts of the leaves of Melodorum fruticosum. The structure of melodamide A (1) was established with a combination of 2D NMR experiments, HR-ESI-MS and X-ray analyses. The other known compounds were identified by comparison of their spectroscopic and physical data with those reported in the literature. Moreover, some isolated compounds were examined for their inhibitory activity towards superoxide anion generation and elastase release in human neutrophils. Among the tested compounds, 1, 3, and 5 exhibited strong inhibition of superoxide anion generation with IC50 values ranging from 5.25 to 8.65 µM. Furthermore, synthesis and biological evaluation of melodamide A (1) and its analogs (14a-p) were described.
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Data mining for signal detection of adverse event safety data.
J Biopharm Stat
PUBLISHED: 01-22-2013
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The Adverse Event Reporting System (AERS) is the primary database designed to support the Food and Drug Administration (FDA) postmarketing safety surveillance program for all approved drugs and therapeutic biologic products. Most current disproportionality analysis focuses on the detection of potential adverse events (AE) involving a single drug and a single AE only. In this paper, we present a data mining biclustering technique based on the singular value decomposition to extract local regions of association for a safety study. The analysis consists of collection of biclusters, each representing an association between a set of drugs with the corresponding set of adverse events. Significance of each bicluster can be tested using disproportionality analysis. Individual drug-event combination can be further tested. A safety data set consisting of 193 drugs with 8453 adverse events is analyzed as an illustration.
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Comparing the response rates for superiority, noninferiority and equivalence testing with multiple-to-one matched binary data.
J Biopharm Stat
PUBLISHED: 01-22-2013
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Paired and multiple-to-one matched data have often been collected in clinical trials and epidemiological safety studies. When the response is binary, the sample size and power are determined by the discordant pairs or matched sets. Fixed sample size determination in assessing response rate difference of superiority, noninferiority, and equivalence tests of paired binary data has been discussed by Lu and Bean (1995)Nam (1997), and Liu et al. (2002). We extend the results of Lu and Bean (1995)Nam (1997), and Liu et al. (2002) to more general cases of multiple-to-one matched binary data. We further examine two issues regarding such tests. First, we examine the issue of simultaneous test and two-stage test for both superiority and noninferiority/equivalence hypotheses. Although, as discussed in Nam (1997) and Liu et al. (2002), the standard errors restricted to null hypothesis are different between superiority and noninferiority test, the monotonic property of the two tests makes the simultaneous testing and switching between the hypotheses valid. Second, in practice, the joint distribution of matched responses is often unknown, and thus determining the sample size using only the background information of the control group could be inefficient. Furthermore, for noninferiority or equivalence tests, the sample sizes are often determined using the unrealistic alternative hypothesis that the response rates of both treatments are identical. We propose to use a two-stage adaptive design strategy for sample size reestimation that uses the interim information to improve the efficiency.
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Bayesian approach to assay sensitivity analysis of thorough QT trials.
J Biopharm Stat
PUBLISHED: 01-22-2013
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One of the analyses recommended in ICH E14 Guidance (International Conference on Harmonisation, 2005) after the test drug is shown to be negative in QT interval prolongation after subjects treated with the test drug is an assay sensitivity analysis of a positive control drug with known effect on QT prolongation. The assay sensitivity is validated if the response profile is shown to be expected and the QT interval after administration of the positive control drug is shown to be at least 5 ms more than placebo. The negative result of the test treatment is validated if the prolongation of the positive control is verified among the study subjects. One of the most frequently used positive control drugs in thorough QT (TQT) trials is moxifloxacin. In order to improve the efficiency of the study and to reduce the number of subjects exposed to moxifloxacin, we explore the potential sample size reduction with a Bayesian approach to the assay sensitivity utilizing the data of historical TQT trials. We derived the distribution of moxifloxacin-induced QT prolongation based on 14 crossover trials and six parallel trials. The estimated distribution is used as a prior distribution to assess the posterior probability that the moxifloxacin-induced QT prolongation is larger than 5 ms. Sample size based on such Bayesian approach will be compared with the conventional frequentist approach for efficiency assessment.
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Sample size of thorough QTc clinical trial adjusted for multiple comparisons.
J Biopharm Stat
PUBLISHED: 01-22-2013
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A thorough QT trial is typically designed to test for two sets of hypotheses. The primary set of hypotheses is for demonstrating that the test treatment will not prolong QT interval. The second set of hypotheses is to demonstrate the assay sensitivity of the positive control treatment in the study population. Both analyses require multiple comparisons by testing the treatment difference measured repeatedly at multiple selected time points. Tsong and Zhong (2010) indicated that for prolongation testing, this involves an intersection-union test that leads to the reduction of study power. It requires type II error rate adjustment in order to maintain proper sample size and power of the test. Tsong et al. (2010) indicated also that the assay sensitivity analysis is carried out using a union-intersection test that leads to the inflation of the family-wise type I error rate. Type I error rate adjustment is required to control the family-wise type I error rate. Zhang and Machado (2008) proposed the sample size calculation of test-placebo QT response difference based on simulation with a multivariate normal distribution model. Even though the results are generally used as guidance for sample size determination for balanced arm TQT trials, they are limited in generalization to various advanced and adaptive designs of TQT trials (Zhang, 2011 ; Tsong, 2013). In this article, we propose a power equation based on multivariate normal distribution of TQT trials. Sample sizes of various TQT designs can be obtained through numerical iteration of the equation.
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On the designs of thorough QT/QTc clinical trials.
J Biopharm Stat
PUBLISHED: 01-22-2013
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A thorough QT trial is typically designed to test for two set of hypotheses. The primary set of hypotheses is for demonstrating that the test treatment will not prolong QT interval. The second set of hypotheses is to demonstrate the assay sensitivity of the positive control treatment in the study population. The conventional designs of thorough QT studies are parallel arms and crossover trials. Although the sample size requirements, the time points of analysis, and the durations of drug administration to reach a steady stage are all different between the test and positive control treatments, the trials are often designed with equal study duration and sample size for the different treatment arms or periods. In order to reduce the number of subjects needed for the parallel-arm thorough QT clinical trial, unconventional designs were proposed and used in the recent years. In this article, we review and discuss a few of the designs and propose some additional designs in the form of unbalanced, group sequential, and adaptive designs with the objective to improve trial efficiency or to reduce the risk of unnecessarily exposure of subjects to the potentially harmful positive control treatment.
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A sensitivity analysis of the modified chi-square ratio statistic for equivalence testing of aerodynamic particle size distribution.
AAPS J
PUBLISHED: 01-04-2013
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Demonstration of equivalence in aerodynamic particle size distribution (APSD) is one key component for establishing bioequivalence of orally inhaled drug products. We previously proposed a modified version of the Chi-square ratio statistic (mCSRS) for APSD equivalence testing and demonstrated that the median of the distribution of the mCSRS (MmCSRS) is a robust metric when test (T) and reference (R) cascade impactor (CI) profiles are identical. Here, we systematically evaluate the behavior of the MmCSRS when T and R CI profiles differ from each other in their mean deposition and variability on a single and multiple sites. All CI profiles were generated by Monte-Carlo simulations based upon modified actual CI data. Twenty thousand sets of 30 T and 30 R CI profiles were simulated for each scenario, and the behavior of the MmCSRS was correlated to metrics that characterize the difference between T and R product in mean deposition and variability. The two key findings were, first, that the MmCSRS is more sensitive to difference between T and R CI profiles on high deposition sites, and second, that a cut-off value for APSD equivalence testing based on the MmCSRS needs to be scaled on the variability of the R product. The former is considered as beneficial for equivalence testing of CI profiles as it decreases the likelihood of failing identical CI profiles by chance, in part, due to increasing analytical variability associated with lower deposition sites. The latter is expected to be important for consistently being able to discriminate equivalent from inequivalent CI profiles.
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Osthole attenuates hepatic injury in a rodent model of trauma-hemorrhage.
PLoS ONE
PUBLISHED: 01-01-2013
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Recent evidences show that osthole possesses anti-inflammatory properties and protective effects following shock-like states, but the mechanism of these effects remains unknown. The p38 mitogen-activated protein kinase (p38 MAPK) pathway exerts anti-inflammatory effects in injury. The aim of this study was to investigate whether p38 MAPK plays any role in the osthole-mediated attenuation of hepatic injury after trauma-hemorrhage. Male Sprague-Dawley rats underwent trauma-hemorrhage (mean blood pressure maintained at approximately 35-40 mmHg for 90 minutes), followed by fluid resuscitation. During resuscitation, a single dose of osthole (3 mg/kg, intravenously) with and without a p38 MAPK inhibitor SB-203580 (2 mg/kg, intravenously), SB-203580 or vehicle was administered. Plasma alanine aminotransferase (ALT) with aspartate aminotransferase (AST) concentrations and various hepatic parameters were measured (n?=?8 rats/group) at 24 hours after resuscitation. The results showed that trauma-hemorrhage increased hepatic myeloperoxidase activity, intercellular adhesion molecule-1 and interleukin-6 levels, and plasma ALT and AST concentrations. These parameters were significantly improved in the osthole-treated rats subjected to trauma-hemorrhage. Osthole treatment also increased hepatic phospho-p38 MAPK expression compared with vehicle-treated trauma-hemorrhaged rats. Co-administration of SB-203580 with osthole abolished the osthole-induced beneficial effects on the above parameters and hepatic injury. These results suggest that the protective effect of osthole administration on alleviation of hepatic injury after trauma-hemorrhage, which is, at least in part, through p38 MAPK-dependent pathway.
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Honokiol dimers and magnolol derivatives with new carbon skeletons from the roots of Magnolia officinalis and their inhibitory effects on superoxide anion generation and elastase release.
PLoS ONE
PUBLISHED: 01-01-2013
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Two honokiol dimers, houpulins A and B (1 and 2), and two magnolol derivatives, houpulins C and D (3 and 4), were isolated and characterized from an ethanol extract obtained from the roots of Magnolia officinalis. The chemical structures were determined based on spectroscopic and physicochemical analyses, which included 1D and 2D NMR, as well as mass spectrometry data. These four oligomers possess new carbon skeletons postulated to be biosynthesized from the coupling of three or four C6-C3 subunits. In addition, the new oligomers were evaluated for inhibition of superoxide anion generation and elastase release, and houpulin B (2) was identified as a new anti-inflammatory lead compound.
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Assessment of reasons for not intensifying antihypertensive treatment in the Taiwanese population.
J. Formos. Med. Assoc.
PUBLISHED: 12-23-2011
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Despite availability of effective antihypertensives, blood pressure (BP) control is usually inadequate. The Reasons for not Intensifying Antihypertensive Treatment (RIAT) registry evaluated the reasons behind not modifying treatment in an international, cross-sectional study in 16 countries.
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EMR2 receptor ligation modulates cytokine secretion profiles and cell survival of lipopolysaccharide-treated neutrophils.
Chang Gung Med J
PUBLISHED: 11-01-2011
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Epidermal growth factor (EGF)-like module-containing mucin-like hormone receptor-like 2 (EMR2) is an adhesion G protein-coupled receptor previously shown to potentiate neutrophil responses to a number of inflammatory stimuli. EMR2 activation promotes neutrophil adhesion and migration, and augments production of reactive oxygen species and degranulation. In this study, we examined the effect of EMR2 ligation by its specific antibody on the cytokine expression profile and cell survival of lipopolysaccharide (LPS)-treated neutrophils.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.