JoVE Visualize What is visualize?
Stop Reading. Start Watching.
Advanced Search
Stop Reading. Start Watching.
Regular Search
Find video protocols related to scientific articles indexed in Pubmed.
KAPtain in charge of multiple missions: Emerging roles of KAP1.
World J Biol Chem
PUBLISHED: 11-28-2014
Show Abstract
Hide Abstract
KAP1/TRIM28/TIF1? was identified nearly twenty years ago as a universal transcriptional co-repressor because it interacts with a large KRAB-containing zinc finger protein (KRAB-ZFP) transcription factor family. Many studies demonstrate that KAP1 affects gene expression by regulating the transcription of KRAB-ZFP-specific loci, trans-repressing as a transcriptional co-repressor or epigenetically modulating chromatin structure. Emerging evidence suggests that KAP1 also functions independent of gene regulation by serving as a SUMO/ubiquitin E3 ligase or signaling scaffold protein to mediate signal transduction. KAP1 is subjected to multiple post-translational modifications (PTMs), including serine/tyrosine phosphorylation, SUMOylation, and acetylation, which coordinately regulate KAP1 function and its protein abundance. KAP1 is involved in multiple aspects of cellular activities, including DNA damage response, virus replication, cytokine production and stem cell pluripotency. Moreover, knockout of KAP1 results in embryonic lethality, indicating that KAP1 is crucial for embryonic development and possibly impacts a wide-range of (patho)physiological manifestations. Indeed, studies from conditional knockout mouse models reveal that KAP1-deficiency significantly impairs vital physiological processes, such as immune maturation, stress vulnerability, hepatic metabolism, gamete development and erythropoiesis. In this review, we summarize and evaluate current literatures involving the biochemical and physiological functions of KAP1. In addition, increasing studies on the clinical relevance of KAP1 in cancer will also be discussed.
Related JoVE Video
Production of Water-Soluble Few-Layer Graphene Mesosheets by Dry Milling with Hydrophobic Drug.
Langmuir
PUBLISHED: 11-20-2014
Show Abstract
Hide Abstract
A novel, fast and easy mechano-chemistry-based (dry milling) method has been developed to exfoliate graphene with hydrophobic drugs generating few layer graphene mesosheets (< 10 nm in thickness and ~ 1 µm in width). The electronic properties of the graphitic structure were partially preserved after the milling treatment compared to Graphene Oxide (GO) prepared by Hummers' method. Several characterization techniques such as thermogravimetric analysis (TGA), Raman spectroscopy, atomic force microscopy (AFM), Electron Microscopy (EM) and molecular dynamics simulation were used to characterize this material. The drug-exfoliated mesosheets were pharmacologically inactive offering a new approach for making water-soluble few-layer graphene mesosheets upon dry milling with hydrophobic drugs, mainly used as exfoliating agents.
Related JoVE Video
Phenethyl isothiocyanate inhibits in vivo growth of subcutaneous xenograft tumors of human malignant melanoma A375.S2 cells.
In Vivo
PUBLISHED: 09-06-2014
Show Abstract
Hide Abstract
Numerous studies have shown that phenethyl isothiocyanate (PEITC) induces apoptosis of different types of human cancer cell lines, however, there are no reports showing that PEITC inhibits tumor growth in a xenograft model of melanoma in nude mice. We investigated effects of PEITC on the growth of xenografted A375.S2 cell tumors in nude BALB/c mice. A375.S2 cancer cells were inoculated subcutaneously into the lower flanks of mice. Seven days post-inoculation, mice having one palpable tumor were randomly divided into three groups and injected intraperitoneally with PEITC (0, 20 and 40 mg/kg). PEITC reduced tumor weight but total body weight was unaffected. These in vivo results provide support for further investigations to determine the potential use of PEITC as an anticancer drug.
Related JoVE Video
Gene expression profiling of sesaminol triglucoside and its tetrahydrofuranoid metabolites in primary rat hepatocytes.
Int J Food Sci Nutr
PUBLISHED: 08-26-2014
Show Abstract
Hide Abstract
Abstract Sesaminol triglucoside is a major lignin in sesame meal and has a methylenedioxyphenyl group and multiple functions in vivo. As a tetrahydrofurofuran type lignan, sesaminol triglucoside is metabolized to mammalian lignans. This investigation studies the effect of sesaminol triglucoside and its tetrahydrofuranoid metabolites (sesaminol, 2-episesaminol, hydroxymethyl sesaminol-tetrahydrofuran, enterolactone, and enterodiol) on gene expression in primary rat hepatocytes using a DNA microarray. Sesame lignans significantly affected the expression of xenobiotic-induced transcripts of cytochrome P450, solute carrier (SLC), and ATP-binding cassette (ABC) transporters. Changes in gene expression were generally greater in response to metabolites with methylenedioxyphenyl moieties (sesaminol triglucoside, sesaminol, and 2-episesaminol) than to the tetrahydrofuranoid metabolites (hydroxymethyl sesaminol-tetrahydrofuran, enterolactone, and enterodiol). Tetrahydrofuran lignans, such as sesaminol triglucoside, sesamin, hydroxymethyl sesaminol-tetrahydrofuran, and sesaminol changed the expression of ABC transporters.
Related JoVE Video
Development and application of a comparative fatty acid analysis method to investigate voriconazole-induced hepatotoxicity.
Clin. Chim. Acta
PUBLISHED: 08-20-2014
Show Abstract
Hide Abstract
As fatty acids play an important role in biological regulation, the profiling of fatty acid expression has been used to discover various disease markers and to understand disease mechanisms. This study developed an effective and accurate comparative fatty acid analysis method using differential labeling to speed up the metabolic profiling of fatty acids.
Related JoVE Video
Pharmacokinetic profile and first preliminary clinical evaluation of bendamustine in Taiwanese patients with heavily pretreated indolent B-cell non-Hodgkin lymphoma and mantle cell lymphoma.
Hematol Oncol
PUBLISHED: 08-11-2014
Show Abstract
Hide Abstract
Prior studies found bendamustine is efficacious in patients with indolent B-cell non-Hodgkin lymphoma (NHL). To date, no studies have reported the efficacy of bendamustine in a Chinese population. This multicentre phase II trial evaluated the pharmacokinetics (PK), safety and efficacy of bendamustine monotherapy in Chinese patients in Taiwan with pretreated indolent B-cell NHL or mantle cell lymphoma (MCL). For PK assessments, patients were randomized (n?=?16; 11 with indolent B-cell NHL and five with MCL) to 90 or 120?mg/m(2) of bendamustine for the first cycle. Plasma levels of bendamustine and its two metabolites were analyzed. For efficacy and safety evaluations, bendamustine 120?mg/m(2) was given to all patients every 3?weeks starting at cycle 2 for a minimum of a total of six cycles. The median age of patients was 61.7?years, and the majority were men (75%). The median number of prior treatments was 4 (range, 1-9 regimens), and all patients were previously treated with rituximab. Bendamustine plasma concentration peaked near the end of infusion and was rapidly eliminated with a mean elimination half-life (t1/2 ) of 0.67-0.8?h. Of the evaluable patients (n?=?14), the overall response rate was 78.6%, including 7.2% of patients having a complete response. Mean progression-free survival was 27.5?weeks. The most common grade 3-4 adverse events were leucopenia (56.3%), neutropenia (56.3%) and thrombocytopenia (25%). In conclusion, bendamustine was efficacious and well tolerated in Taiwanese patients with indolent NHL and MCL with a similar PK profile to that of other populations. Copyright © 2014 John Wiley & Sons, Ltd.
Related JoVE Video
Computed tomography angiography provides limited benefit in the evaluation of patients with pelvic fractures.
Am J Emerg Med
PUBLISHED: 08-01-2014
Show Abstract
Hide Abstract
Computed tomography angiography (CTA) has been applied in imaging studies for the assessment of most abdominal and pelvic injuries in some trauma centers. However, in most institutions, CTA is not routinely performed as part of the computed tomography scan protocol. In this study, we aimed to assess the efficiency of CTA in the evaluation of patients with pelvic fractures.
Related JoVE Video
Long-Term Electrophysiological and Behavioral Analysis on the Improvement of Visual Working Memory Load, Training Gains, and Transfer Benefits.
J Behav Brain Sci
PUBLISHED: 07-22-2014
Show Abstract
Hide Abstract
Recent evidence demonstrates that with training, one can enhance visual working memory (VWM) capacity and attention over time in the near transfer tasks. Not only do these studies reveal the characteristics of VWM load and the influences of training, they may also provide insights into developing effective rehabilitation for patients with VWM deficiencies. However, few studies have investigated VWM over extended periods of time and evaluated transfer benefits on non-trained tasks. Here, we combined behavioral and electroencephalographical approaches to investigate VWM load, training gains, and transfer benefits. Our results reveal that VWM capacity is directly correlated to the difference of event-related potential waveforms. In particular, the "magic number 4" can be observed through the contralateral delay amplitude and the average capacity is 3.25-item over 15 participants. Furthermore, our findings indicate that VWM capacity can be improved through training; and after training exercises, participants from the training group are able to dramatically improve their performance. Likewise, the training effects on non-trained tasks can also be observed at the 12th week after training. Therefore, we conclude that participants can benefit from training gains, and augmented VWM capacity sustained over long periods of time on specific variety of tasks.
Related JoVE Video
Carbamazepine attenuates inducible nitric oxide synthase expression through Akt inhibition in activated microglial cells.
Pharm Biol
PUBLISHED: 07-15-2014
Show Abstract
Hide Abstract
Abstract Background: Carbamazepine, which was developed primarily for the treatment of epilepsy, is now also useful for the treatment of non-epileptic disorders and inflammatory hyperalgesia. However, the mechanism of its anti-neuroinflammatory action remains poorly understood.
Related JoVE Video
An arginine-rich motif of ring finger protein 4 (RNF4) oversees the recruitment and degradation of the phosphorylated and SUMOylated Krüppel-associated box domain-associated protein 1 (KAP1)/TRIM28 protein during genotoxic stress.
J. Biol. Chem.
PUBLISHED: 06-06-2014
Show Abstract
Hide Abstract
Krüppel-associated box domain-associated protein 1 (KAP1) is a universal transcriptional corepressor that undergoes multiple posttranslational modifications (PTMs), including SUMOylation and Ser-824 phosphorylation. However, the functional interplay of KAP1 PTMs in regulating KAP1 turnover during DNA damage response remains unclear. To decipher the role and cross-talk of multiple KAP1 PTMs, we show here that DNA double strand break-induced KAP1 Ser-824 phosphorylation promoted the recruitment of small ubiquitin-like modifier (SUMO)-targeted ubiquitin E3 ligase, ring finger protein 4 (RNF4), and subsequent RNF4-mediated, SUMO-dependent degradation. Besides the SUMO interacting motif (SIM), a previously unrecognized, but evolutionarily conserved, arginine-rich motif (ARM) in RNF4 acts as a novel recognition motif for selective target recruitment. Results from combined mutagenesis and computational modeling studies suggest that RNF4 utilizes concerted bimodular recognition, namely SIM for Lys-676 SUMOylation and ARM for Ser(P)-824 of simultaneously phosphorylated and SUMOylated KAP1 (Ser(P)-824-SUMO-KAP1). Furthermore, we proved that arginines 73 and 74 within the ARM of RNF4 are required for efficient recruitment to KAP1 or accelerated degradation of promyelocytic leukemia protein (PML) under stress. In parallel, results of bimolecular fluorescence complementation assays validated the role of the ARM in recognizing Ser(P)-824 in living cells. Taken together, we establish that the ARM is required for RNF4 to efficiently target Ser(P)-824-SUMO-KAP1, conferring ubiquitin Lys-48-mediated proteasomal degradation in the context of double strand breaks. The conservation of such a motif may possibly explain the requirement for timely substrate selectivity determination among a myriad of SUMOylated proteins under stress conditions. Thus, the ARM dynamically regulates the SIM-dependent recruitment of targets to RNF4, which could be critical to dynamically fine-tune the abundance of Ser(P)-824-SUMO-KAP1 and, potentially, other SUMOylated proteins during DNA damage response.
Related JoVE Video
Minimizing the total service time of discrete dynamic berth allocation problem by an iterated greedy heuristic.
ScientificWorldJournal
PUBLISHED: 05-26-2014
Show Abstract
Hide Abstract
Berth allocation is the forefront operation performed when ships arrive at a port and is a critical task in container port optimization. Minimizing the time ships spend at berths constitutes an important objective of berth allocation problems. This study focuses on the discrete dynamic berth allocation problem (discrete DBAP), which aims to minimize total service time, and proposes an iterated greedy (IG) algorithm to solve it. The proposed IG algorithm is tested on three benchmark problem sets. Experimental results show that the proposed IG algorithm can obtain optimal solutions for all test instances of the first and second problem sets and outperforms the best-known solutions for 35 out of 90 test instances of the third problem set.
Related JoVE Video
Concise syntheses of 7-anilino-indoline-N-benzenesulfonamides as antimitotic and vascular disrupting agents.
Bioorg. Med. Chem.
PUBLISHED: 05-19-2014
Show Abstract
Hide Abstract
Described herein is the development of a novel series of 7-anilino-indoline-N-benzenesulfonamides, derived from ABT751 (1), as potent anticancer agents. Amongst the synthesized series, compounds 6, 12, 13, and 14 have shown comparable to better anticancer activity on comparing with compound 1. 7-(4-Cyanophenylamino)-1-(4-methoxybenzenesulfonyl)indoline (13) was found to be the most potent one with up to 6 fold better activity against KB, HT29, and MKN45 cancer cell lines with IC50 values of 49.7, 149, and 92nM, respectively. Compound 13 was also found inhibiting multidrug resistant cancer cell lines, blocking cell cycle at G2/M phase, and inhibiting tubulin polymerization. Capillary disruption assay results revealed that compound 13 was able to disrupt formed capillaries in a concentration-dependent manner without affecting cell viability.
Related JoVE Video
Azaindolylsulfonamides, with a more selective inhibitory effect on histone deacetylase 6 activity, exhibit antitumor activity in colorectal cancer HCT116 cells.
J. Med. Chem.
PUBLISHED: 05-06-2014
Show Abstract
Hide Abstract
A series of indolylsulfonylcinnamic hydroxamates has been synthesized. Compound 12, (E)-3-(3-((1H-pyrrolo[2,3-b]pyridin-1-yl)sulfonyl)phenyl)-N-hydroxyacrylamide, which has a 7-azaindole core cap, was shown to have antiproliferative activity against KB, H460, PC3, HSC-3, HONE-1, A549, MCF-7, TSGH, MKN45, HT29, and HCT116 human cancer cell lines. Pharmacological studies indicated that 12 functions as a potent HDAC inhibitor with an IC50 value of 0.1 ?M. It is highly selective for histone deacetylase 6 (HDAC6) and is 60-fold more active than against HDAC1 and 223-fold more active than against HDAC2. It has a good pharmacokinetic profile with oral bioavailability of 33%. In in vivo efficacy evaluations in colorectal HCT116 xenografts, compound 12 suppresses tumor growth more effectively than SAHA (1, N-hydroxy-N'-phenyloctanediamide) and is therefore seen as a suitable candidate for further investigation.
Related JoVE Video
Hypotension does not always make computed tomography scans unfeasible in the management of blunt abdominal trauma patients.
Injury
PUBLISHED: 05-03-2014
Show Abstract
Hide Abstract
Computed tomography (CT) scans have been used worldwide to evaluate patients with blunt abdominal trauma (BAT). However, CT scans have traditionally been considered to be a part of a secondary survey that can only be performed after the patient's haemodynamics have stabilised. In this study, we attempted to evaluate the role of the CT scan in managing BAT patients with hypotension.
Related JoVE Video
Antimitotic and vascular disrupting agents: 2-hydroxy-3,4,5-trimethoxybenzophenones.
Eur J Med Chem
PUBLISHED: 02-26-2014
Show Abstract
Hide Abstract
2-Hydroxy-3,4,5-trimethoxybenzophenones (8-16) manifest pseudo-ring formation involving intramolecular hydrogen bonding of the 2-OH and the carbonyl group. Among the synthetic products described in this report, (3-hydroxy-4-methoxyphenyl)(2-hydroxy-3,4,5-trimethoxyphenyl)-methanone (14) and (3-amino-4-methoxyphenyl)(2-hydroxy-3,4,5-trimethoxy-phenyl)methanone (16) exhibit significant antiproliferative activity against KB cells with IC50 values of 11.1 and 11.3 nM, respectively. These two compounds also displayed tubulin affinity comparable to that of combretastatin A-4. In studies with human umbilical vein endothelial cells, compounds 14 and 16 revealed concentration-dependent vascular-disrupting properties. The results support the rationale of the pseudo-ring concept and suggest further investigation of A-ring modification in these benzophenones.
Related JoVE Video
Improvements in hand function after intensive bimanual training are not associated with corticospinal tract dysgenesis in children with unilateral cerebral palsy.
Exp Brain Res
PUBLISHED: 02-19-2014
Show Abstract
Hide Abstract
Unilateral cerebral palsy (CP) results from damage to the developing brain that occurs within the first 2 years of life. Previous studies found associations between asymmetry in the size of the corticospinal tract (CST) from the two hemispheres and severity of hand impairments in children with unilateral CP. The extent to which CST damage affects the capacity for hand function improvement is unknown. This study examines the association between an estimate of CST dysgenesis and (1) hand function and (2) the efficacy of intensive bimanual training in improving hand function. Children with unilateral CP, age 3.6-14.9 years, n = 35, received intensive bimanual training. Children engaged in bimanual functional/play activities (6 h/day, 15 days). Peduncle asymmetry, an estimate of CST dysgenesis, was measured on T1-weighted magnetic resonance imaging scans. Hand function was measured pre- and post-treatment using the assisting hand assessment (AHA) and Jebsen-Taylor test of hand function (JTTHF). AHA and JTTHF improved post-treatment (p < 0.001). Peduncle asymmetry was correlated with baseline AHA and JTTHF (p < 0.001) but not with AHA or JTTHF improvement post-training (R(2) < 0.1, p > 0.2). An estimate of CST dysgenesis is correlated with baseline hand function but is a poor predictor of training efficacy, possibly indicating a flexibility of developing motor systems to mediate recovery.
Related JoVE Video
Quantification of target analytes in various biofluids using a postcolumn infused-internal standard method combined with matrix normalization factors in liquid chromatography-electrospray ionization mass spectrometry.
J Chromatogr A
PUBLISHED: 01-26-2014
Show Abstract
Hide Abstract
Liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) has become one of the most widely used methods in pharmaceutical laboratories. Although LC-ESI-MS provides high sensitivity and high specificity for quantifying target analytes in complicated biofluids, the associated severe matrix effects (MEs) generally result in large quantification errors. Here, we propose a novel strategy for correcting MEs in various biofluids using a postcolumn infused-internal standard (PCI-IS) method in combination with matrix normalization factors (MNFs). We used the MNFs to normalize the encountered MEs in various biofluids to the MEs encountered in standard solutions. The use of a postcolumn infused-internal standard also corrects the MEs for individual samples. When using the PCI-IS method in combination with MNFs, the calibration curve generated from standard solutions can be applied to quantify the target analytes in various biofluids. We applied this new approach to quantify etoposide and etoposide catechol in plasma and CSF. The accuracy of the test results showed that over 93% of the data have quantification errors less than 20% and that 99% of the data have quantification errors less than 30%. The successful application of this method to evaluate real clinical samples revealed that our proposed MNFs in combination with the PCI-IS method largely simplifies the entire method development and validation processes, saves a great deal of time and cost without sacrificing quantification accuracy, and provides a simple means of quantifying target analytes in various biofluids. This method will be particularly useful in fields in which the same target analytes need to be quantified in various types of matrices, including bioanalysis, forensic toxicology, environmental studies, and food safety control.
Related JoVE Video
Intra-abdominal injury is easily overlooked in the patients with concomitant unstable hemodynamics and pelvic fractures.
Am J Emerg Med
PUBLISHED: 01-25-2014
Show Abstract
Hide Abstract
Transcatheter arterial embolization (TAE) is usually necessary in the management of hemodynamically unstable patients with concomitant pelvic fractures. Given the critical conditions of such patients, TAE is at times performed only according to the results of a primary evaluation without computed tomographic (CT) imaging. Therefore, the evaluation of associated intra-abdominal injuries (IAIs) might be insufficient. Clinically, some patients have required post-TAE laparotomy due to further deterioration. In this study, we attempted to determine a feasible protocol for post-TAE observation.
Related JoVE Video
Using a postcolumn-infused internal standard for correcting the matrix effects of urine specimens in liquid chromatography-electrospray ionization mass spectrometry.
J Chromatogr A
PUBLISHED: 01-08-2014
Show Abstract
Hide Abstract
Matrix effects (MEs) are a major problem affecting the quantitative accuracy of liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) when analyzing complicated samples. While analyzing urine specimens, the wide diversity of endogenous materials and different urine concentrations may result in inaccurate quantification. In this study, we propose a postcolumn-infused internal standard (PCI-IS) strategy for universal correction of MEs in urine specimens. MEs can be effectively corrected by dividing the target analyte signal intensity by the PCI-IS intensity. To evaluate the performance of PCI-IS, we used 6 benzodiazepine (BZD) drugs in 5 different concentrations of urine matrixes as a test model. The divergence of the BZD drug signal responses in 5 different urine matrixes was expressed using their respective coefficients of variation (CV) to evaluate the efficiency of using PCI-IS in correcting matrix effects. The CV of the BZD drug signal intensities in these 5 different concentrations of the urine matrixes were reduced from 10 to 30% to less than 10% when the PCI-IS correction method was employed. When the PCI-IS method was used to correct the 6 BZDs in 25 real human urine samples, over 90% of the test results exhibited quantification errors of less than 20%, and all of the test results had quantification errors of less than 30%. These results demonstrate that the PCI-IS method can resolve the problem of inaccurate quantification that arises from the diversity of urine specimens. The PCI-IS method is particularly useful for clinical analysis or forensic toxicology to improve the quantification accuracy in an economical way.
Related JoVE Video
Screening and management of major bile leak after blunt liver trauma: a retrospective single center study.
Scand J Trauma Resusc Emerg Med
PUBLISHED: 01-07-2014
Show Abstract
Hide Abstract
Major bile leak after blunt liver trauma is rare but challenging. It usually requires endoscopic retrograde cholangiography (ERC) for management. However, there is still lack of specific indications. The aim of this study is to elucidate risk factors for major bile leak and indications for early ERC after blunt liver trauma.
Related JoVE Video
Analysis of changes in transcription start site distribution by a classification approach.
Gene
PUBLISHED: 01-07-2014
Show Abstract
Hide Abstract
Change in transcription start site (TSS) usage is an important mechanism for the control of transcription process, and has a significant effect on the isoforms being transcribed. One of the goals in the study of TSS is the understanding of how and why their usage differs in different tissues or under different conditions. In light of recent efforts in the mapping of transcription start site landscape using high-throughput sequencing approaches, a quantitative and automated method is needed to process all the data that are being produced. In this work we propose a statistical approach that will classify changes in TSS distribution between different samples into several categories of changes that may have biological significance. Genes selected by the classifiers can then be analyzed together with additional supporting data to determine their biological significance. We use a set of time-course TSS data from mouse dendritic cells stimulated with lipopolysaccharide (LPS) to demonstrate the usefulness of our method.
Related JoVE Video
The impact of spectral resolution on the mismatch response to Mandarin Chinese tones: an ERP study of cochlear implant simulations.
Clin Neurophysiol
PUBLISHED: 01-07-2014
Show Abstract
Hide Abstract
This study examined the impact of spectral resolution on the processing of lexical tones and the number of frequency channels required for a cochlear implant (CI) to transmit Chinese tonal information to the brain.
Related JoVE Video
Localizing movement-related primary sensorimotor cortices with multi-band EEG frequency changes and functional MRI.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Electroencephalographic (EEG) oscillations in multiple frequency bands can be observed during functional activity of the cerebral cortex. An important question is whether activity of focal areas of cortex, such as during finger movements, is tracked by focal oscillatory EEG changes. Although a number of studies have compared EEG changes to functional MRI hemodynamic responses, we can find no previous research that relates the fMRI hemodynamic activity to localization of the multiple EEG frequency changes observed in motor tasks. In the present study, five participants performed similar thumb and finger movement tasks in parallel EEG and functional MRI studies. We examined changes in five frequency bands (from 5-120 Hz) and localized them using 256 dense-array EEG (dEEG) recordings and high-resolution individual head models. These localizations were compared with fMRI localizations in the same participants. Results showed that beta-band (14-30 Hz) desynchronizations (power decreases) were the most robust effects, appearing in all individuals, consistently localized to the hand region of the primary motor cortex, and consistently aligned with fMRI localizations.
Related JoVE Video
Complicated intra-abdominal infections worldwide: the definitive data of the CIAOW Study.
World J Emerg Surg
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
The CIAOW study (Complicated intra-abdominal infections worldwide observational study) is a multicenter observational study underwent in 68 medical institutions worldwide during a six-month study period (October 2012-March 2013). The study included patients older than 18 years undergoing surgery or interventional drainage to address complicated intra-abdominal infections (IAIs). 1898 patients with a mean age of 51.6 years (range 18-99) were enrolled in the study. 777 patients (41%) were women and 1,121 (59%) were men. Among these patients, 1,645 (86.7%) were affected by community-acquired IAIs while the remaining 253 (13.3%) suffered from healthcare-associated infections. Intraperitoneal specimens were collected from 1,190 (62.7%) of the enrolled patients. 827 patients (43.6%) were affected by generalized peritonitis while 1071 (56.4%) suffered from localized peritonitis or abscesses. The overall mortality rate was 10.5% (199/1898). According to stepwise multivariate analysis (PR?=?0.005 and PE?=?0.001), several criteria were found to be independent variables predictive of mortality, including patient age (OR?=?1.1; 95%CI?=?1.0-1.1; p?
Related JoVE Video
TMPRSS6 rs855791 polymorphism influences the susceptibility to iron deficiency anemia in women at reproductive age.
Int J Med Sci
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Genome-wide-association studies have identified the TMPRSS6 polymorphism rs855791 has the strongest association with red blood cell indices or iron parameters in general population. Whether this genetic variant influences the susceptibility of iron deficiency anemia (IDA) in women with menstruation has not been well studied.
Related JoVE Video
Gastric metastasis of lung cancer mimicking an adrenal tumor.
Case Rep Gastroenterol
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Lung cancer is one of the leading causes of cancer deaths worldwide. Metastatic spreads of lung cancer are often found in the adrenal glands, bone, liver, brain and kidneys; the gastrointestinal tract is less commonly involved. However, according to some reports in the literature, the incidence of gastrointestinal metastases, most of them asymptomatic, might be as frequent as 11% in autopsy studies of lung cancer, which suggests that this condition is not as rare as it was previously considered. We report a very rare case of small cell lung cancer with a solitary gastric metastasis mimicking an adrenal tumor which was belatedly diagnosed due to its unusual presentation and treated actively with surgery and chemotherapy, achieving a relatively favorable outcome.
Related JoVE Video
Therapeutic drug monitoring and pharmacogenetic study of HIV-infected ethnic Chinese receiving efavirenz-containing antiretroviral therapy with or without rifampicin-based anti-tuberculous therapy.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Plasma efavirenz concentrations in HIV-infected patients with tuberculosis (TB) may be affected by cytochrome P450 (CYP) 2B6 single-nucleotide polymorphisms and concurrent rifampicin use. We aimed to investigate the effects of CYP2B6 G516T polymorphisms and concomitant rifampicin use on the plasma efavirenz concentrations in HIV-infected Taiwanese.
Related JoVE Video
Comparison of Structured Skill and Unstructured Practice During Intensive Bimanual Training in Children With Unilateral Spastic Cerebral Palsy.
Neurorehabil Neural Repair
PUBLISHED: 12-27-2013
Show Abstract
Hide Abstract
. High-intensity training aims to improve hand function in children with unilateral spastic cerebral palsy (USCP). However, the extent to which skill training is required is not known.
Related JoVE Video
Metabolomics of Ginger Essential Oil against Alcoholic Fatty Liver in Mice.
J. Agric. Food Chem.
PUBLISHED: 11-12-2013
Show Abstract
Hide Abstract
Fatty liver is significantly associated with hepatic cirrhosis and liver cancer. Excessive alcohol consumption causes alcoholic fatty liver disease (AFLD). Ginger has been reported to exhibit antioxidant potential and hepatoprotective activity. In the present study, a mouse model for AFLD was developed by employing male C57BL/6 mice that were fed an alcohol-containing liquid diet (Lieber-DeCarli diet) ad libitum. In the treatment groups, ginger essential oil (GEO) and citral were orally administered every day for 4 weeks. Serum biochemical analysis, antioxidant enzyme activity analysis, and histopathological evaluation revealed that GEO and citral exhibited hepatoprotective activity against AFLD. Metabolites in serum samples were profiled by high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS). Metabolomic data indicated the amounts of metabolites such as d-glucurono-6,3-lactone, glycerol-3-phosphate, pyruvic acid, lithocholic acid, 2-pyrocatechuic acid, and prostaglandin E1 were increased after alcohol administration, but the levels were recovered in treatment groups. The analysis indicated that ginger possesses hepatoprotective properties against AFLD. Furthermore, these metabolites can serve as early noninvasive candidate biomarkers in the clinical application of AFLD for health management.
Related JoVE Video
Linking transcriptional changes over time in stimulated dendritic cells to identify gene networks activated during the innate immune response.
PLoS Comput. Biol.
PUBLISHED: 11-01-2013
Show Abstract
Hide Abstract
The innate immune response is primarily mediated by the Toll-like receptors functioning through the MyD88-dependent and TRIF-dependent pathways. Despite being widely studied, it is not yet completely understood and systems-level analyses have been lacking. In this study, we identified a high-probability network of genes activated during the innate immune response using a novel approach to analyze time-course gene expression profiles of activated immune cells in combination with a large gene regulatory and protein-protein interaction network. We classified the immune response into three consecutive time-dependent stages and identified the most probable paths between genes showing a significant change in expression at each stage. The resultant network contained several novel and known regulators of the innate immune response, many of which did not show any observable change in expression at the sampled time points. The response network shows the dominance of genes from specific functional classes during different stages of the immune response. It also suggests a role for the protein phosphatase 2a catalytic subunit ? in the regulation of the immunoproteasome during the late phase of the response. In order to clarify the differences between the MyD88-dependent and TRIF-dependent pathways in the innate immune response, time-course gene expression profiles from MyD88-knockout and TRIF-knockout dendritic cells were analyzed. Their response networks suggest the dominance of the MyD88-dependent pathway in the innate immune response, and an association of the circadian regulators and immunoproteasomal degradation with the TRIF-dependent pathway. The response network presented here provides the most probable associations between genes expressed in the early and the late phases of the innate immune response, while taking into account the intermediate regulators. We propose that the method described here can also be used in the identification of time-dependent gene sub-networks in other biological systems.
Related JoVE Video
Furanylazaindoles: potent anticancer agents in vitro and in vivo.
J. Med. Chem.
PUBLISHED: 10-09-2013
Show Abstract
Hide Abstract
Preliminary biological data on 7-anilino-6-azaindoles (8-11) suggested that hydrophobic substituents at C7 contribute to enhancement of antiproliferative activity. A novel series of 7-aryl-6-azaindole-1-benzenesulfonamides (12-22) were developed and showed improved cytotoxicity compared to ABT751 (5). The conversion of C7 phenyl rings into C7 heterocycles led to a remarkable improvement of antiproliferative activity. Among all the synthetic products, 7-(2-furanyl)-1-(4-methoxybenzenesulfonyl)-6-azaindole (21) exhibited the most potent anticancer activity against KB, HT29, MKN45, and H460 cancer cell lines with IC50 values of 21.1, 32.0, 27.5, and 40.0 nM, respectively. Bioassays indicated that 21 not only inhibits tubulin polymerization by binding to tubulin at the colchicine binding site but also arrests the cell cycle at the G2/M phase with slight arrest at the sub-G1 phase. Compound 21 also functions as a vascular disrupting agent and dose-dependently inhibits tumor growth without significant change of body weight in an HT29 xenograft mouse model. Taken together, compound 21 has potential for further development as a novel class of anticancer agents.
Related JoVE Video
Screening and confirmation of 62 drugs of abuse and metabolites in urine by ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry.
J Anal Toxicol
PUBLISHED: 09-30-2013
Show Abstract
Hide Abstract
An ultra-high-performance liquid chromatography--quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) method for the screening and confirmation of 62 drugs of abuse and their metabolites in urine was developed in this study. The most commonly abused drugs, including amphetamines, opioids, cocaine, benzodiazepines (BZDs) and barbiturates, and many other new and emerging abused drugs, were selected as the analytes for this study. Urine samples were diluted 5-fold with deionized water before analysis. Using a superficially porous micro-particulate column and an acetic acid-based mobile phase, 54 basic and 8 acidic analytes could be detected within 15 and 12 min in positive and negative ionization modes, respectively. The MS collision energies for the 62 analytes were optimized, and their respective fragmentation patterns were constructed in the in-house library for confirmatory analysis. The coefficients of variation of the intra- and inter-day precision of the analyte responses all were <17.39%. All analytes, except barbital, showed matrix effects of 77-121%. The limits of detection of the 62 analytes were between 2.8 and 187.5 ng/mL, which were lower than their respective cut-off concentrations (20-500 ng/mL). Ten urine samples from patients undergoing methadone treatment were analyzed by the developed UHPLC-QTOF-MS method, and the results were compared with the immunoassay method.
Related JoVE Video
Tyrosol and its analogues inhibit alpha-melanocyte-stimulating hormone induced melanogenesis.
Int J Mol Sci
PUBLISHED: 09-18-2013
Show Abstract
Hide Abstract
Melanin is responsible for skin color and plays a major role in defending against harmful external factors such as ultraviolet (UV) irradiation. Tyrosinase is responsible for the critical steps of melanogenesis, including the rate-limiting step of tyrosine hydroxylation. The mechanisms of action of skin hypopigmenting agents are thought to be based on the ability of a given agent to inhibit the activity of tyrosinase and, hence, down regulate melanin synthesis. Tyrosol and its glycoside, salidroside, are active components of Rhodiola rosea, and in our preliminary study we found that Rhodiola rosea extract inhibited melanogenesis. In this study, we examined the effects of tyrosol and its analogues on melanin synthesis. We found that treatment of B16F0 cells to tyrosol (1), 4-hydroxyphenylacetic acid (5), 3-hydroxyphenylacetic acid (6), 2-hydroxyphenylacetic acid (7), or salidroside (11) resulted in a reduction in melanin content and inhibition of tyrosinase activity as well as its expression. Tyrosol (1), 4-hydroxyphenylacetic acid (5) and 2-hydroxyphenylacetic acid (7) suppressed MC1R expression. Tyrosol (1), 4-hydroxyphenylacetic acid (5), 3-hydroxyphenylacetic acid (6), and 2-hydroxyphenylacetic acid (7) inhibited ?-MSH induced TRP-1 expression, but salidroside (11) did not. All the compounds did not affect MITF and TRP-2 expression. Furthermore, we found that the cell viability of tyrosol (1), 4-hydroxyphenylacetic acid (5), 3-hydroxyphenylacetic acid (6), and 2-hydroxyphenylacetic acid (7) at concentrations below 4 mM and salidroside (11) at concentrations below 0.5 mM were higher than 90%. The compounds exhibited metal-coordinating interactions with copper ion in molecular docking with tyrosinase. Our results suggest that tyrosol, 4-hydroxyphenylacetic acid, 3-hydroxyphenylacetic acid, 2-hydroxyphenylacetic acid, and salidroside are potential hypopigmenting agents.
Related JoVE Video
Field assessment of Orientia tsutsugamushi infection in small mammals and its association with the occurrence of human scrub typhus in Taiwan.
Acta Trop.
PUBLISHED: 08-13-2013
Show Abstract
Hide Abstract
We conducted an extensive study in Taiwan of Orientia tsutsugamushi (OT) infection in small wild mammals. Field trapping was carried out at six districts in eastern and western Taiwan as well as various offshore islands during the period 2006-2010. A total of 1061 specimens representing 11 rodent species were captured. The presence of OT infection was assessed by indirect immunofluorescence assay and polymerase chain reaction assays of 56-kDa type-specific antigen gene. The chigger infestation rate among the animals was 35% (371/1061). Among these, OT was detected in 64% (238/371) of the chiggers from the infested animals and in the spleens from 273 (34.3%) of 797 animals. Excluding animals in the Suncus murinus group, the antibody positive rate of scrub typhus was 69.1% (477 of 690 of serum samples). The prevalence of OT infection in animals from areas with a low incidence of human cases of scrub typhus was significantly lower than that in rodents obtained from regions with a high incidence of human cases of the disease (44.4%±4.0% vs. 71.2%±9.7%, p<0.001). In Taiwan, the prevalence of OT infection in wild rodents is considerably high and appears to correlate positively with the occurrence of scrub typhus in humans.
Related JoVE Video
Hepatoprotective effects of Ixora parviflora extract against exhaustive exercise-induced oxidative stress in mice.
Molecules
PUBLISHED: 08-13-2013
Show Abstract
Hide Abstract
Ixora parviflora, a species of the Rubiaceae, is rich in polyphenols and flavonoids, and has been traditionally used as a folk medicine. An I. parviflora extract (IPE) has great antioxidant activity in vitro, including a scavenging effect on superoxide radicals, reducing power, and ferrous ion-chelating ability. However, whether IPE is efficacious against oxidative damage in vivo is not known. The purpose of this study was to determine the protective effects of IPE treatment on hepatic oxidative stress and antioxidant defenses after exhaustive exercise in mice. Fifty male C57BL/6 mice (6 week old) were randomly divided into five groups and designated a sedentary control with vehicle (C), and exhaustive exercise with vehicle (IPE0), low dosage (IPE10), medium dosage (IPE50) and high dosage (IPE100) of IPE at 0, 10, 50, and 100 mg/kg, respectively. After a single bout of exhaustive swimming exercise challenge, levels of blood ammonia and creatine kinase (CK), and hepatic superoxide dismutase (SOD) protein expression, thiobarbituric acid-reactive substance (TBARS), and gp91(phox), p22(phox), and p47(phox) subunits of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase expressions in the IPE0 group were significantly affected compared to those of the C group, but they were all significantly inhibited by the IPE treatments. Results of the present in vivo study in mice indicate that I. parviflora extract possesses antioxidative and hepatoprotective potential following exhaustive exercise.
Related JoVE Video
Intentional forgetting reduces the semantic processing of to-be-forgotten items: an ERP study of item-method directed forgetting.
Psychophysiology
PUBLISHED: 08-05-2013
Show Abstract
Hide Abstract
In two ERP experiments, we examined whether active inhibition is involved in intentional forgetting. Both experiments consisted of a nondirected-forgetting (nDF) and a directed-forgetting (DF) block. Participants were sequentially presented with a prime, an R/F (remember/forget) cue, and a target. Participants made lexical decisions to both the primes and targets (Experiment 1) or only to the targets (Experiment 2). They were also instructed to remember or to forget the primes in response to the R/F cues in the DF block but to ignore these cues in the nDF block. The N400 semantic priming effect was observed when comparing the ERPs elicited by semantically unrelated and related targets in the DF block. In comparison to the nDF block, the N400 effect was greatly reduced for targets preceded by F cues in the DF block. These findings suggest that semantic processing is reduced by the instruction to forget and active inhibition is involved in intentional forgetting.
Related JoVE Video
The diminishing role of pelvic x-rays in the management of patients with major torso injuries.
Am J Emerg Med
PUBLISHED: 07-30-2013
Show Abstract
Hide Abstract
A pelvic x-ray (PXR) can be used as an effective screening tool to evaluate pelvic fractures and stability. However, associated intra-abdominal/retroperitoneal organ injuries and hemorrhage should also be considered and evaluated in patients with major torso injuries. An abdominal/pelvic computed tomographic (CT) scan may provide higher resolution and more information than a PXR. The role of conventional PXRs was delineated in the current study in the context of the development of the CT scan.
Related JoVE Video
Application of quantitative estimates of fecal hemoglobin concentration for risk prediction of colorectal neoplasia.
World J. Gastroenterol.
PUBLISHED: 06-30-2013
Show Abstract
Hide Abstract
To determine the role of the fecal immunochemical test (FIT), used to evaluate fecal hemoglobin concentration, in the prediction of histological grade and risk of colorectal tumors.
Related JoVE Video
Pelvic circumferential compression devices benefit patients with pelvic fractures who need transfers.
Am J Emerg Med
PUBLISHED: 06-19-2013
Show Abstract
Hide Abstract
Patients with pelvic fracture usually require transfers to trauma centers for additional advanced treatment. Patient safety during the transfer should always be a priority. The noninvasive pelvic circumferential compression device (PCCD) can reportedly provide a tamponade effect, which reduces hemorrhage. In the present study, we evaluated the feasibility and efficiency of PCCD in patients with pelvic fracture who required transfer to trauma centers.
Related JoVE Video
Propofol induces DNA damage in mouse leukemic monocyte macrophage RAW264.7 cells.
Oncol. Rep.
PUBLISHED: 05-28-2013
Show Abstract
Hide Abstract
Propofol is one of the most widely clinically used intravenous anesthetic, and it induces apoptosis in human and murine leukemia cell lines. Yet, whether propofol causes DNA damage and affects the mRNA expression of repair-associated genes in cancer cells remains undetermined. In the present study, we investigated the effects of propofol on DNA damage and associated mRNA gene expression in RAW264.7 cells. Comet assay and DNA gel electrophoresis were used to evaluate DNA damage in RAW264.7 cells and propofol-inhibited cell growth in vitro. The results revealed a longer DNA tail and DNA fragmentation. Real-time PCR assay was used to examine mRNA gene expression of DNA damage and DNA repair-associated genes. Following exposure to propofol for 48 h, a decrease in the mRNA expression of DNA-PK, BRCA1, MGMT and p53 was noted in the RAW264.7 cells. Results from the western blotting indicated that p53, MGMT, 14-3-3-?, BRCA1 and MDC1 proteins were decreased while p-p53 and p-H2A.X(S140) were increased in the RAW264.7 cells following exposure to propofol. In conclusion, exposure to propofol caused DNA damage and inhibited mRNA expression and protein levels of repair-associated genes in RAW264.7 cells.
Related JoVE Video
Association of blood lead and mercury with estimated GFR in herbalists after the ban of herbs containing aristolochic acids in Taiwan.
Occup Environ Med
PUBLISHED: 05-23-2013
Show Abstract
Hide Abstract
This study was undertaken to explore the association of estimated glomerular filtration rate (GFR) with exposure to aristolochic acids (ALAs) and nephrotoxic metals in herbalists after the ban of herbs containing ALAs in Taiwan.
Related JoVE Video
Analgesic and anti-inflammatory bioactivities of eburicoic acid and dehydroeburicoic acid isolated from Antrodia camphorata on the inflammatory mediator expression in mice.
J. Agric. Food Chem.
PUBLISHED: 05-16-2013
Show Abstract
Hide Abstract
Eburicoic acid (TR1) and dehydroeburicoic acid (TR2), an active ingredient from Antrodia camphorata (AC) solid-state culture, were evaluated for analgesic and anti-inflammatory effects. Treatment with TR1 and TR2 significantly inhibited a number of acetic acid-induced writhing responses and formalin-induced pain in the late phase. In the anti-inflammatory test, TR1 and TR2 decreased paw edema at the fourth and fifth hour after ?-carrageenan (Carr) administration and increased the activities of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) in the paw edema tissue. We also demonstrated that TR1 and TR2 significantly attenuated the malondialdehyde (MDA), nitric oxide (NO), tumor necrosis factor (TNF-?), and interleukin-1? (IL-1?) levels in either edema paw or serum at the fifth hour after Carr injection. Western blotting revealed that TR1 and TR2 decreased Carr-induced inducible nitric oxide synthase (iNOS) and cycloxyclase (COX-2) expressions at the fifth hour in paw edema. Treatment with TR1 and TR2 also diminished neutrophil infiltration into the paw edema at the fifth hour. The present study suggests that the anti-inflammatory mechanisms of TR1 and TR2 might be related to the decrease of inflammatory cytokines and an increase of antioxidant enzyme activity.
Related JoVE Video
Comparative Proteomic Analysis of Rodent Plasma and Mesenteric Lymph.
Chin J Physiol
PUBLISHED: 05-10-2013
Show Abstract
Hide Abstract
The lymph has long been considered as the plasma filtrate and the proteomes of the lymph have received scanty attention. Currently, mesenteric lymph is reported to play an important role in the pathogenesis of multiple organ dysfunction syndrome in some critical illnesses. A better understanding of the composition and proteomes of mesenteric lymph becomes imperative to disclose the mechanistic role of mesenteric lymph. Seven male Sprague-Dawley rats were fasted overnight, and anesthetized to collect plasma and mesenteric lymph. The specimens were subjected to proteomic analysis using twodimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). An average of 434 and 412 protein spots were found in the gels of the plasma and mesenteric lymph respectively. Peptide mass fingerprint analysis identified 77 proteins for 212 protein spots. The 2-DE proteomic pattern of mesenteric lymph was largely similar to that of the plasma. As in the plasma, large protein spots of albumin dominated the protein pattern in mesenteric lymph. Other major proteins identified in 2-DE gels included immunoglobulin heavy and light chains, fibrinogen ?-, ?- and ?-chains, serotransferrin, protease inhibitors, kininogens, macroglobulins, haptoglobin, and apolipoproteins. Meanwhile, mesenteric lymph contained an array of proteins that differentiated it from the plasma. The most differentially expressed proteins in mesenteric lymph were ?-fibrinogen, protease inhibitors, and proteins related to lipid transport/metabolism. The study presents a detailed description of mesenteric lymph proteomes of a common experimental animal in physiological status using a common proteomic approach. These results provide the basis for future research.
Related JoVE Video
MPT0B098, a novel microtubule inhibitor that destabilizes the hypoxia-inducible factor-1? mRNA through decreasing nuclear-cytoplasmic translocation of RNA-binding protein HuR.
Mol. Cancer Ther.
PUBLISHED: 04-25-2013
Show Abstract
Hide Abstract
Microtubule inhibitors have been shown to inhibit hypoxia-inducible factor-1? (HIF-1?) expression through inhibition translation or enhancing protein degradation. Little is known of the effect of microtubule inhibitors on the stability of HIF-1? mRNA. We recently discovered a novel indoline-sulfonamide compound, 7-aryl-indoline-1-benzene-sulfonamide (MPT0B098), as a potent microtubule inhibitor through binding to the colchicine-binding site of tubulin. MPT0B098 is active against the growth of various human cancer cells, including chemoresistant cells with IC50 values ranging from 70 to 150 nmol/L. However, normal cells, such as human umbilical vein endothelial cells (HUVEC), exhibit less susceptibility to the inhibitory effect of MPT0B098 with IC50 of 510 nmol/L. Similar to typical microtubule inhibitors, MPT0B098 arrests cells in the G2-M phase and subsequently induces cell apoptosis. In addition, MPT0B098 effectively suppresses VEGF-induced cell migration and capillary-like tube formation of HUVECs. Distinguished from other microtubule inhibitors, MPT0B098 not only inhibited the expression levels of HIF-1? protein but also destabilized HIF-1? mRNA. The mechanism of causing unstable of HIF-1? mRNA by MPT0B098 is through decreasing RNA-binding protein, HuR, translocation from the nucleus to the cytoplasm. Notably, MPT0B098 effectively suppresses tumor growth and microvessel density of tumor specimens in vivo. Taken together, our results provide a novel mechanism of inhibiting HIF-1? of a microtubule inhibitor MPT0B098. MPT0B098 is a promising anticancer drug candidate with potential for the treatment of human malignancies.
Related JoVE Video
Simultaneous quantification of antimicrobial agents for multidrug-resistant bacterial infections in human plasma by ultra-high-pressure liquid chromatography-tandem mass spectrometry.
Talanta
PUBLISHED: 04-23-2013
Show Abstract
Hide Abstract
Antibiotic-resistant bacterial infection is one of the most serious clinical problems worldwide. Vancomycin, teicoplanin, daptomycin, and colistin are glycopeptide and lipopeptide antibiotics that are frequently used to treat multidrug-resistant bacterial infections. Therapeutic drug monitoring is recommended to ensure both safety and efficacy and to improve clinical outcomes. This study developed a fast, simple, and sensitive ultra-high-pressure liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous determination of the concentrations of these four drugs in human plasma. The sample preparation process includes a simple protein denaturation step using acetonitrile, followed by an 11-fold dilution with 0.1% formic acid. Eight target peaks for the four drugs can be analyzed within 3 min using a Kinetex™ 2.6 ?m C18 column. The mass spectrometry parameters were optimized, and two transitions for each target peak were used for multiple reaction monitoring, which provided high sensitivity and specificity. The UHPLC-MS/MS method was validated over clinical concentration ranges. The intra-day and inter-day precisions for the ratio of the peak area of each analyte to the peak area of the internal standard were all below 12.7 and 14.7% relative standard deviations, respectively. The accuracy at low, medium, and high concentrations of the eight target peaks was between 89.3 and 110.7%. The standard curves for the analytes were linear and had coefficients of determination higher than 0.997. The limits of detection were all below 70 ng mL(-1). The use of this method to analyze patient plasma samples confirmed that it is effective for the therapeutic drug monitoring of these four drugs and can be used to improve the therapeutic efficacy and safety of treatment with antibiotics.
Related JoVE Video
Gene expression profiling for analysis acquired oxaliplatin resistant factors in human gastric carcinoma TSGH-S3 cells: the role of IL-6 signaling and Nrf2/AKR1C axis identification.
Biochem. Pharmacol.
PUBLISHED: 04-23-2013
Show Abstract
Hide Abstract
Oxaliplatin treatment is a mainstay of treatment for advanced gastrointestinal tract cancer, but the underlying mechanisms of acquired oxaliplatin resistance remain largely obscured. We previously demonstrated that increased DNA repair capacity and copper-transporting ATPase 1 (ATP7A) level contributed to oxaliplatin resistance in the human gastric carcinoma cell line TSGH-S3 (S3). In the present study, we applied gene array technology to identify additional resistance factors in S3 cells. We found that interleukin-6 (IL-6), aldo-keto reductase 1C1 (AKR1C1), and AKR1C3 are the top 3 upregulated genes in S3 cells when compared with parent TSGH cells. Despite a higher level of endogenous IL-6 in S3, IL-6 receptor (IR-6R, gp-80, and gp-130) levels were similar between TSGH and S3 cells. The addition of exogenous IL-6, IL-6 targeted siRNA, or neutralizing antibodies neither affected Stat3 activation, a downstream target of IL-6, nor changed oxaliplatin sensitivity in S3 cells. However, manipulation of AKR1C activity with siRNA or AKR1C inhibitors significantly reversed oxaliplatin resistance. AKR1Cs are classical antioxidant response element (ARE) genes that can be transcriptionally upregulated by nuclear factor erythroid 2-related factor 2 (Nrf2). Knockdown of Nrf2 not only decreased the levels of AKR1C1, AKR1C2, and AKR1C3 mRNA and protein but also reversed oxaliplatin resistance in S3 cells. Taken together, these results suggest that activation of the Nrf2/AKR1C axis may contribute to oxaliplatin resistance in S3 cells but that the IL-6 signaling pathway did not contribute to resistance. Manipulation of Nrf2/AKR1Cs activity may be useful for management of oxaliplatin-refractory gastric cancers.
Related JoVE Video
Comparing the outcomes of two strategies for colorectal tumor detection: policy-promoted screening program versus health promotion service.
J Chin Med Assoc
PUBLISHED: 04-18-2013
Show Abstract
Hide Abstract
The Taiwanese government has proposed a population-based colorectal tumor detection program for the average-risk population. This studys objectives were to understand the outcomes of these screening policies and to evaluate the effectiveness of the program.
Related JoVE Video
A novel aerosol-mediated drug delivery system for inner ear therapy: intratympanic aerosol methylprednisolone can attenuate acoustic trauma.
IEEE Trans Biomed Eng
PUBLISHED: 04-15-2013
Show Abstract
Hide Abstract
We developed a novel aerosol-mediated drug delivery system for inner ear therapy by using a silicon-based multiple-Fourier horn nozzle. Intratympanic aerosol (ITA) methylprednisolone (MP) delivery can protect hearing after acoustic trauma. The highest concentration of MP (38.9 ± 5.47 ppm) appeared at 2 h and declined rapidly within 10 h. The concentrations of MP remained at a relatively low level for more than 10 h. Compared to the baseline, the auditory brainstem response (ABR) thresholds shifted markedly at 1 h after noise exposure in all groups (p < 0.05). From the cochleograms, it can be noted that the main lesions encompassed the 2-20 kHz frequency range. Significant differences ( ) were observed for the range between 5 and 8 kHz in the cell loss of outer hair cells (OHCs). The losses for IHCs were lower than for OHCs. The MP movement in the middle ear was simulated by a convection diffusion equation with a relaxation time. The relaxation time was 0.5 h, and the concentration threshold of MP on the round window membrane (RWM) in the middle ear (C T) was 8900 ppm. Using the unit hydrograph (UH) method, we obtained a proper boundary concentration on the RWM at the cochlea, which resulted in a well-fit concentration. Finally, a linking mechanism between the middle ear and the cochlea was established by the RWM. The adjustable permeability and concentration threshold provide the flexibility to match the peak times and peak values of the concentration on the RWM in the middle ear and the cochlea.
Related JoVE Video
HP1 promotes tumor suppressor BRCA1 functions during the DNA damage response.
Nucleic Acids Res.
PUBLISHED: 04-15-2013
Show Abstract
Hide Abstract
The DNA damage response (DDR) involves both the control of DNA damage repair and signaling to cell cycle checkpoints. Therefore, unraveling the underlying mechanisms of the DDR is important for understanding tumor suppression and cellular resistance to clastogenic cancer therapeutics. Because the DDR is likely to be influenced by chromatin regulation at the sites of DNA damage, we investigated the role of heterochromatin protein 1 (HP1) during the DDR process. We monitored double-strand breaks (DSBs) using the ?H2AX foci marker and found that depleting cells of HP1 caused genotoxic stress, a delay in the repair of DSBs and elevated levels of apoptosis after irradiation. Furthermore, we found that these defects in repair were associated with impaired BRCA1 function. Depleting HP1 reduced recruitment of BRCA1 to DSBs and caused defects in two BRCA1-mediated DDR events: (i) the homologous recombination repair pathway and (ii) the arrest of cell cycle at the G2/M checkpoint. In contrast, depleting HP1 from cells did not affect the non-homologous end-joining (NHEJ) pathway: instead it elevated the recruitment of the 53BP1 NHEJ factor to DSBs. Notably, all three subtypes of HP1 seemed to be almost equally important for these DDR functions. We suggest that the dynamic interaction of HP1 with chromatin and other DDR factors could determine DNA repair choice and cell fate after DNA damage. We also suggest that compromising HP1 expression could promote tumorigenesis by impairing the function of the BRCA1 tumor suppressor.
Related JoVE Video
Metabolomic Dynamic Analysis of Hypoxia in MDA-MB-231 and the Comparison with Inferred Metabolites from Transcriptomics Data.
Cancers (Basel)
PUBLISHED: 04-12-2013
Show Abstract
Hide Abstract
Hypoxia affects the tumor microenvironment and is considered important to metastasis progression and therapy resistance. Thus far, the majority of global analyses of tumor hypoxia responses have been limited to just a single omics level. Combining multiple omics data can broaden our understanding of tumor hypoxia. Here, we investigate the temporal change of the metabolite composition with gene expression data from literature to provide a more comprehensive insight into the system level in response to hypoxia. Nuclear magnetic resonance spectroscopy was used to perform metabolomic profiling on the MDA-MB-231 breast cancer cell line under hypoxic conditions. Multivariate statistical analysis revealed that the metabolic difference between hypoxia and normoxia was similar over 24 h, but became distinct over 48 h. Time dependent microarray data from the same cell line in the literature displayed different gene expressions under hypoxic and normoxic conditions mostly at 12 h or earlier. The direct metabolomic profiles show a large overlap with theoretical metabolic profiles deduced from previous transcriptomic studies. Consistent pathways are glycolysis/gluconeogenesis, pyruvate, purine and arginine and proline metabolism. Ten metabolic pathways revealed by metabolomics were not covered by the downstream of the known transcriptomic profiles, suggesting new metabolic phenotypes. These results confirm previous transcriptomics understanding and expand the knowledge from existing models on correlation and co-regulation between transcriptomic and metabolomics profiles, which demonstrates the power of integrated omics analysis.
Related JoVE Video
Isoflavonoid-Rich Flemingia macrophylla Extract Attenuates UVB-Induced Skin Damage by Scavenging Reactive Oxygen Species and Inhibiting MAP Kinase and MMP Expression.
Evid Based Complement Alternat Med
PUBLISHED: 03-26-2013
Show Abstract
Hide Abstract
In this study, we investigated the antioxidant activity and anti-photoaging properties of an extract of Flemingia macrophylla, a plant rich in isoflavonoid content. Pretreatment of fibroblasts with Flemingia macrophylla extract (FME) inhibited elastase activity, promoted the protein expression of type I procollagen, and attenuated the phosphorylation of mitogen-activated protein (MAP) kinase and the protein expression of matrix-metalloproteinase- (MMP-) 1, 3, and 9. The IC50 values were 2.1? ? g/mL for DPPH radical scavenging ability, 366.8? ? g/mL for superoxide anion scavenging ability, 178.9? ? g/mL for hydrogen peroxide scavenging ability, and 230.9? ? g/mL for hydroxyl radical scavenging ability. Also, exposure of erythrocytes to various concentrations of FME (50-500? ? g/mL) resulted in a dose- and time-dependent inhibition of AAPH-induced hemolysis. In human fibroblasts, FME at 10? ? g/mL was shown to be a potent scavenger of UV-induced reactive oxygen species (ROS). The antioxidant and anti-photoaging properties of FME make it an ideal anti-intrinsic aging and anti-photoaging agent.
Related JoVE Video
Phyto-SERM constitutes from Flemingia macrophylla.
Int J Mol Sci
PUBLISHED: 03-15-2013
Show Abstract
Hide Abstract
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).
Related JoVE Video
Plasma metabolomic profiles predict near-term death among individuals with lower extremity peripheral arterial disease.
J. Vasc. Surg.
PUBLISHED: 02-19-2013
Show Abstract
Hide Abstract
Individuals with peripheral arterial disease (PAD) have a nearly two-fold increased risk of all-cause and cardiovascular disease mortality compared to those without PAD. This pilot study determined whether metabolomic profiling can accurately identify patients with PAD who are at increased risk of near-term mortality.
Related JoVE Video
Intruder-induced asymmetry in compartmentalized granular gases.
Phys Rev E Stat Nonlin Soft Matter Phys
PUBLISHED: 02-08-2013
Show Abstract
Hide Abstract
The clustering behavior of a compartmentalized monodisperse granular gas with the addition of one heavy intruding particle is investigated experimentally. Depending on the number of particles, the presence of a heavy intruder leads to three population states: a homogeneous state, an expelled clustering state, and a fully clustering state. These states are found to be consistent with the clustering of a purely monodisperse granular gas in an asymmetric compartmentalized structure. We obtain an exact relation between the size of an intruder and the elevation of the compartment bottom. This relation quantifies the particle-expelling ability of a heavy intruder, and suggests that the one-intruder system is a type of asymmetric system with an intruder-size-related asymmetrical index ?. Under the framework of the flux model, a ?-associated ? function is proposed to quantitatively reproduce the experimental results.
Related JoVE Video
Histone deacetylase inhibition improved cardiac functions with direct antifibrotic activity in heart failure.
Int. J. Cardiol.
PUBLISHED: 02-08-2013
Show Abstract
Hide Abstract
Histone deacetylases (HDACs), important epigenetic regulatory enzymes, can reduce cardiac hypertrophy and cardiac fibrosis. However, the mechanisms underlying the antifibrotic activity of HDAC inhibitors remain unclear. The purposes of this study were to evaluate the effects of an HDAC inhibitor on systolic heart failure (HF) and investigate the potential mechanisms.
Related JoVE Video
A novel synthetic microtubule inhibitor, MPT0B214 exhibits antitumor activity in human tumor cells through mitochondria-dependent intrinsic pathway.
PLoS ONE
PUBLISHED: 02-08-2013
Show Abstract
Hide Abstract
Agents that interfere with mitotic progression by disturbing microtubule dynamics are commonly used for cancer treatment. Previously, a series of aroylquinolone regioisomers as novel microtubule inhibitors were discovered. One of these new compounds, MPT0B214 inhibited tubulin polymerization through strongly binding to the tubulins colchicine-binding site and had cytotoxic activity in a variety of human tumor cell lines. After treatment with MPT0B214, KB cells were arrested in the G2-M phase before cell death occurred, which were associated with upregulation of cyclin B1, dephosphorylation of Cdc2, phosphorylation of Cdc25C and elevated expression of the mitotic marker MPM-2. Furthermore, the compound induced apoptotic cell death through mitochondria/caspase 9-dependent pathway. Notably, several KB-derived multidrug-resistant cancer cell lines were also sensitive to MPT0B214 treatment. These findings showed that MPT0B214 is a potential compound in the treatment of various malignancies.
Related JoVE Video
Active Constituents from Liriope platyphylla Root against Cancer Growth In Vitro.
Evid Based Complement Alternat Med
PUBLISHED: 02-06-2013
Show Abstract
Hide Abstract
Liriope spicata is a well-known herb in traditional Chinese medicine, and its root has been clinically demonstrated to be effective in the treatment of metabolic and neural disorders. The constituents isolated from Liriope have also recently been shown to possess anticancer activity, although the mechanism of which remains largely unknown. Here, we illustrate the anticancer activity of LPRP-9, one of the active fractions we fractionated from the Liriope platyphylla root part (LPRP) extract. Treatment with LPRP-9 significantly inhibited proliferation of cancer cell lines MCF-7 and Huh-7 and down-regulated the phosphorylation of AKT. LPRP-9 also activates the stress-activated MPAK, JNK, p38 pathways, the p53 cell-cycle checkpoint pathway, and a series of caspase cascades while downregulating expression of antiapoptotic factors Bcl-2, Bcl-XL, and survivin. Such activities strongly suggest a role for LPRP-9 in apoptosis and autophagy. We further purified and identified the compound (-)-Liriopein B from LPRP-9, which is capable of inhibiting AKT phosphorylation at low concentration. The overall result highlights the anticancer property of LPRP-9, suggests its mechanism for inhibition of proliferation and promotion of cell death for cancer cells via regulation of multitarget pathways, and denotes the importance of purifying components of fraction LPRP-9 to aid cancer therapy.
Related JoVE Video
True ion pick (TIPick): a denoising and peak picking algorithm to extract ion signals from liquid chromatography/mass spectrometry data.
J Mass Spectrom
PUBLISHED: 02-05-2013
Show Abstract
Hide Abstract
Liquid Chromatography-Time of Flight Mass Spectrometry has become an important technique for toxicological screening and metabolomics. We describe TIPick a novel algorithm that accurately and sensitively detects target compounds in biological samples. TIPick comprises two main steps: background subtraction and peak picking. By subtracting a blank chromatogram, TIPick eliminates chemical signals of blank injections and reduces false positive results. TIPick detects peaks by calculating the S(CC(INI)) values of extracted ion chromatograms (EICs) without considering peak shapes, and it is able to detect tailing and fronting peaks. TIPick also uses duplicate injections to enhance the signals of the peaks and thus improve the peak detection power. Commonly seen split peaks caused by either saturation of the mass spectrometer detector or a mathematical background subtraction algorithm can be resolved by adjusting the mass error tolerance of the EICs and by comparing the EICs before and after background subtraction. The performance of TIPick was tested in a data set containing 297 standard mixtures; the recall, precision and F-score were 0.99, 0.97 and 0.98, respectively. TIPick was successfully used to construct and analyze the NTU MetaCore metabolomics chemical standards library, and it was applied for toxicological screening and metabolomics studies.
Related JoVE Video
Hydroalcoholic extract of Rhodiola rosea L. (Crassulaceae) and its hydrolysate inhibit melanogenesis in B16F0 cells by regulating the CREB/MITF/tyrosinase pathway.
Food Chem. Toxicol.
PUBLISHED: 02-04-2013
Show Abstract
Hide Abstract
We investigated the effects of an aqueous alcohol extract of Rhodiola rosea (R. rosea) and its hydrolysate on melanin synthesis and the mechanisms mediating the activity. The ratio of tyrosol to salidroside was 2.3 in hydroalcoholic extract, and 51.0 in hydrolysate. We found that R. rosea extract and its hydrolysate inhibited melanin synthesis and tyrosinase activity in mouse melanoma cells (B16F0 cells). R. rosea extract also inhibited gene and protein expression of melanocortin 1 receptor (MC1R) and inhibited c-AMP response element binding protein (CREB) phosphorylation, suppressed the activation of AKT and glycogen synthase kinase-3 beta (GSK3?), and inhibited the expression of microphthalmia-associated transcription factor (MITF) and tyrosinase-related protein 1 (TRP-1). R. rosea hydrolysate inhibited the phosphorylation of CREB, the activation of AKT and GSK3?, and the expression of MITF and tyrosinase. Our results suggest that R. rosea extract is a novel tyrosinase inhibitor and that it exerts its effects by regulating the CREB/MITF/tyrosinase pathway in B16F0. Further in vivo studies are needed to determine the effectiveness of R. rosea extract as a skinwhitening agent.
Related JoVE Video
Heart failure and angiotensin II modulate atrial Pitx2c promotor methylation.
Clin. Exp. Pharmacol. Physiol.
PUBLISHED: 01-21-2013
Show Abstract
Hide Abstract
Heart failure (HF) can increase atrial fibrillation and induce cardiac hypermethylation. The homeobox gene Pitx2c plays important roles in the genesis of atrial fibrillation and the promoter region of Pitx2c contains cytosine-phosphate-guanine islands. Therefore, epigenetic modification by hypermethylation may reduce Pitx2c expression in atrial myocytes. The aim of the present study were to evaluate whether HF can modulate DNA methylation of Pitx2c and the potential mechanisms involved. We used real-time polymerase chain reaction, immunoblotting and pyrosequencing to investigate RNA and protein expression, as well as the methylation of Pitx2c, in isoproterenol-induced HF, healthy rat left atria and in HL-1 cells with and without (control) exposure to angiotensin (Ang) II (0.1 and 1 ?mol/L) or isoproterenol (1 or 10 ?mol/L) for 24 h. The HF atrium exhibited increased Pitx2c promoter methylation with increased DNA methyltransferase (DNMT) 1 and decreased Pitx2c protein levels compared with the normal atrium. Angiotensin II (0.1 and 1 ?mol/L), increased Pitx2c promoter methylation in HL-1 cells with increased DNMT1 and decreased Pitx2c and Kir2.1 protein levels compared with control cells. These effects were attenuated by the methylation inhibitor 5-aza-2-deoxycytidine (0.1 ?mol/L) and by the AngII receptor blocker losartan (10 ?mol/L). However, isoproterenol (1 and 10 ?mol/L) did not change the expression of the Pitx2c, DNMT1 and Kir2.1 proteins. In conclusion, HF induces Pitx2c promoter hypermethylation and AngII may contribute to the hypermethylation in HF.
Related JoVE Video
Metabolomic characterization of rhubarb species by capillary electrophoresis and ultra-high-pressure liquid chromatography.
Electrophoresis
PUBLISHED: 01-20-2013
Show Abstract
Hide Abstract
This study developed CE and ultra-high-pressure LC (UHPLC) methods coupled with UV detectors to characterize the metabolomic profiles of different rhubarb species. The optimal CE conditions used a BGE with 15 mM sodium tetraborate, 15 mM sodium dihydrogen phosphate monohydrate, 30 mM sodium deoxycholate, and 30% ACN v/v at pH 8.3. The optimal UHPLC conditions used a mobile phase composed of 0.05% phosphate buffer and ACN with gradient elution. The gradient profile increased linearly from 10 to 21% ACN within the first 25 min, then increased to 33% ACN for the next 10 min. It took another 5 min to reach the 65% ACN, then for the next 5 min, it stayed unchanged. Sixteen samples of Rheum officinale and Rheum tanguticum collected from various locations were analyzed by CE and UHPLC methods. The metabolite profiles of CE were aligned and baseline corrected before chemometric analysis. Metabolomic signatures of rhubarb species from CE and UHPLC were clustered using principle component analysis and distance-based redundancy analysis; the clusters were not only able to discriminate different species but also different cultivation regions. Similarity measurements were performed by calculating the correlation coefficient of each sample with the authentic samples. Hybrid rhizome was clearly identified through similarity measurement of UHPLC metabolite profile and later confirmed by gene sequencing. The present study demonstrated that CE and UHPLC are efficient and effective tools to identify and authenticate herbs even coupled with simple detectors.
Related JoVE Video
Metabolomic analysis of complex chinese remedies: examples of induced nephrotoxicity in the mouse from a series of remedies containing aristolochic Acid.
Evid Based Complement Alternat Med
PUBLISHED: 01-08-2013
Show Abstract
Hide Abstract
Aristolochic acid nephropathy is caused by aristolochic acid (AA) and AA-containing herbs. In traditional Chinese medicine, a principle called "Jun-Chen-Zou-Shi" may be utilized to construct a remedial herbal formula that attempts to mitigate the toxicity of the main ingredient. This study used Bu-Fei-A-Jiao-Tang (BFAJT) to test if the compound remedy based on a principle of "Jun-Chen-Zou-Shi" can decrease the toxicity of AA-containing herbs. We compared the three toxicities of AA standard, Madouling (an Aristolochia herb), and a herbal formula BFAJT. AA standard was given for BALB/c mice at a dose of 5?mg/kg?bw/day or 7.5?mg/kg?bw/day for 10 days. Madouling and BFAJT were given at an equivalence of AA 0.5?mg/kg bw/day for 21 days. Nephrotoxicity was evaluated by metabolomics and histopathology. The urinary metabolomics profiles were characterized by (1)H NMR spectroscopy. The spectral data was analyzed with partial least squares discriminant analysis, and the significant differential metabolites between groups were identified. The result showed different degrees of acute renal tubular injuries, and metabolomics analysis found that the kidney injuries were focused in proximal renal tubules. Both metabolomics and pathological studies revealed that AA standard, Madouling, and BFAJT were all nephrotoxicants. The compositions of the compound remedy did not diminish the nephrotoxicity caused by AA.
Related JoVE Video
Neonauclea reticulata (Havil.) Merr Stimulates Skin Regeneration after UVB Exposure via ROS Scavenging and Modulation of the MAPK/MMPs/Collagen Pathway.
Evid Based Complement Alternat Med
PUBLISHED: 01-03-2013
Show Abstract
Hide Abstract
In this study, we investigated whether the protective effects of Neonauclea reticulata water extract against ultraviolet B (UVB) irradiation in human skin fibroblast cell cultures (Hs68) are governed by its ability to protect against oxidative stress and expression of matrix metalloproteinases (MMPs). We found that Neonauclea reticulata extract exhibited DPPH scavenging activity and inhibited AAPH-induced haemolysis of erythrocytes in a dose- and time-dependent manner. We also found that pretreatment of fibroblasts with Neonauclea reticulata water extract resulted in markedly lower levels of MMP-1, -3, and -9 expressions. Furthermore, our results indicate that Neonauclea reticulata extract inhibits the expression of MMPs by inhibiting ERK, JNK, and p38 phosphorylation. Our results also demonstrate that treatment with Neonauclea reticulata extract protects against UVB-induced depletion of collagen. In addition, Neonauclea reticulata extract did not have a cytotoxic effect. These findings indicate that the antioxidant activity of Neonauclea reticulata extract resulted in inhibition of MMP-1, -3, and -9 expressions and in increased levels of collagen activity. Our results suggest that Neonauclea reticulata extract can protect against photoaging.
Related JoVE Video
Complicated intra-abdominal infections in a worldwide context: an observational prospective study (CIAOW Study).
World J Emerg Surg
PUBLISHED: 01-02-2013
Show Abstract
Hide Abstract
Despite advances in diagnosis, surgery, and antimicrobial therapy, mortality rates associated with complicated intra-abdominal infections remain exceedingly high. The World Society of Emergency Surgery (WSES) has designed the CIAOW study in order to describe the clinical, microbiological, and management-related profiles of both community- and healthcare-acquired complicated intra-abdominal infections in a worldwide context. The CIAOW study (Complicated Intra-Abdominal infection Observational Worldwide Study) is a multicenter observational study currently underway in 57 medical institutions worldwide. The study includes patients undergoing surgery or interventional drainage to address complicated intra-abdominal infections. This preliminary report includes all data from almost the first two months of the six-month study period. Patients who met inclusion criteria with either community-acquired or healthcare-associated complicated intra-abdominal infections (IAIs) were included in the study. 702 patients with a mean age of 49.2 years (range 18-98) were enrolled in the study. 272 patients (38.7%) were women and 430 (62.3%) were men. Among these patients, 615 (87.6%) were affected by community-acquired IAIs while the remaining 87 (12.4%) suffered from healthcare-associated infections. Generalized peritonitis was observed in 304 patients (43.3%), whereas localized peritonitis or abscesses was registered in 398 (57.7%) patients.The overall mortality rate was 10.1% (71/702). The final results of the CIAOW Study will be published following the conclusion of the study period in March 2013.
Related JoVE Video
Surface ?-enolase promotes extracellular matrix degradation and tumor metastasis and represents a new therapeutic target.
PLoS ONE
PUBLISHED: 01-01-2013
Show Abstract
Hide Abstract
In previous research, we found ?-enolase to be inversely correlated with progression-free and overall survival in lung cancer patients and detected ?-enolase on the surface of lung cancer cells. Based on these findings, we hypothesized that surface ?-enolase has a significant role in cancer metastasis and tested this hypothesis in the current study. We found that ?-enolase was co-immunoprecipitated with urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor, and plasminogen in lung cancer cells and interacted with these proteins in a cell-free dot blotting assay, which can be interrupted by ?-enolase-specific antibody. ?-Enolase in lung cancer cells co-localized with these proteins and was present at the site of pericellular degradation of extracellular matrix components. Treatment with antibody against ?-enolase in vitro suppressed cell-associated plasminogen and matrix metalloproteinase activation, collagen and gelatin degradation, and cell invasion. Examination of the effect of treatment with shRNA plasmids revealed that down regulation of ?-enolase decreases extracellular matrix degradation by and the invasion capacity of lung cancer cells. Adoptive transfer of ?-enolase-specific antibody to mice resulted in accumulation of antibody in subcutaneous tumor and inhibited the formation of tumor metastasis in lung and bone. This study demonstrated that surface ?-enolase promotes extracellular matrix degradation and invasion of cancer cells and that targeting surface ?-enolase is a promising approach to suppress tumor metastasis.
Related JoVE Video
Coordinate to guard: crosstalk of phosphorylation, sumoylation, and ubiquitylation in DNA damage response.
Front Oncol
PUBLISHED: 11-30-2011
Show Abstract
Hide Abstract
Small ubiquitin-like modifier-1/2/3 (SUMO-1/2/3) and ubiquitin share similar structure and utilize analogous machinery for protein lysine conjugation. Although sumoylation and ubiquitylation have distinct functions, they are often tightly associated with each other to fine-tune protein fate in transducing signals to regulate a wide variety of cellular functions, including DNA damage response, cell proliferation, DNA replication, embryonic development, and cell differentiation. In this Perspective, we specifically highlight the role of sumoylation and ubiquitylation in ataxia-telangiectasia mutated (ATM) signaling in response to DNA double-strand breaks and hypothesize that ATM-induced phosphorylation is a unique node in regulating SUMO-targeted ubiquitylation in mammalian cells to combat DNA damage and to maintain genome integrity. A potential role for the coordination of three types of post-translational modification in dictating the tempo and extent of cellular response to genotoxic stress is speculated.
Related JoVE Video
Knockdown of HURP inhibits the proliferation of hepacellular carcinoma cells via downregulation of gankyrin and accumulation of p53.
Biochem. Pharmacol.
PUBLISHED: 11-24-2011
Show Abstract
Hide Abstract
We determined earlier that the hepatoma upregulated protein (HURP) is overexpressed in hepatocellular carcinoma (HCC), but the role of this protein during cancer development and progression remains unknown. Here, we observed that the overexpression of HURP in HEK293 cells promoted the ubiquitination of p53 and its degradation by the proteasome. In contrast, HURP knockdown using short-hairpin RNA reversed these effects. Knockdown of HURP promoted the accumulation of p53 in SK-Hep-1 cells (p53+/-), and these cells showed reduced proliferation, while the p53-mutant Mahlavu cells were not affected. HURP knockdown did not affect the proliferation of H1299 lung carcinoma cells and Hep3B HCC cells which lack p53. Knockdown of HURP also sensitized SK-Hep-1 cells to cisplatin. On the other hand, the expression of exogenous p53 in H1299 and Hep3B cells was decreased following overexpression of HURP, and these cells showed decreased sensitivity to cisplatin-induced apoptosis. Importantly, overexpression of HURP promoted the proliferation of HEK293 cells in an anchorage-independent manner, and inoculation of SK-Hep-1 cancer cells that expressed short-hairpin RNA to knockdown HURP resulted in smaller tumors in nude mice. Gankyrin, a positive regulator of the E3 ubiquitin ligase MDM2, was found to be upregulated following HURP expression, and gankyrin knockdown decreased the HURP-mediated downregulation of p53. Notably, we detected a positive correlation between elevated HURP and gankyrin protein levels in HCC patients (r(2) = 0.778; N = 9). Taken together, these results indicate that HURP represents an oncogene that may play a role in HCC progression and chemoresistance.
Related JoVE Video
5-Amino-2-aroylquinolines as highly potent tubulin polymerization inhibitors. Part 2. The impact of bridging groups at position C-2.
J. Med. Chem.
PUBLISHED: 11-23-2011
Show Abstract
Hide Abstract
A variety of studies on the modification of combretastatin A-4 triggered our interest in the impact of the linkers between the 3,4,5-trimethoxyphenyl ring and 5-amino-6-methoxyquinoline on biological activity. The replacement of the carbonyl group with bond, amine, ether, sulfide, and sulfone groups was evaluated in this study. The results showed that compounds 14 and 15 containing sulfide and sulfone groups between the 3,4,5-trimethoxyphenyl ring (A-ring) and 5-amino-6-methoxyquinoline exhibited substantial antiproliferative activity against KB, HT29, and MKN45 cells with mean IC50 values of 42 and 12 nM, respectively. 15 inhibited the tubulin polymerization with an IC50 value of 2.0 ?M, similar to that with CA4. The continued work on the C-5 substituents of 3,4,5-trimethoxybenzoyl-6-methoxyquinoline derivatives demonstrated that compound 7 possessing OH at C-5 exhibited excellent antiproliferative activity with mean IC50 values of 3.4 nM and microtubule destabilizing potency with an IC50 of 1.5 ?M, comparable to that of CA4 (IC50=1.9 ?M). It also exhibited substantial vascular disrupting effects. Compounds 7 and 15 exhibited significant efficacy against MDR/MRP-related drug-resistant cell lines (KB-vin10, KB-S15, and KB-7D) with mean IC50 values of 6.7 and 2.6 nM, respectively.
Related JoVE Video

What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

How does it work?

We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

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.