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.
TCRklass: A New K-String-Based Algorithm for Human and Mouse TCR Repertoire Characterization.
J. Immunol.
PUBLISHED: 11-19-2014
Show Abstract
Hide Abstract
The next-generation sequencing technology has promoted the study on human TCR repertoire, which is essential for the adaptive immunity. To decipher the complexity of TCR repertoire, we developed an integrated pipeline, TCRklass, using K-string-based algorithm that has significantly improved the accuracy and performance over existing tools. We tested TCRklass using manually curated short read datasets in comparison with in silico datasets; it showed higher precision and recall rates on CDR3 identification. We applied TCRklass on large datasets of two human and three mouse TCR repertoires; it demonstrated higher reliability on CDR3 identification and much less biased V/J profiling, which are the two components contributing the diversity of the repertoire. Because of the sequencing cost, short paired-end reads generated by next-generation sequencing technology are and will remain the main source of data, and we believe that the TCRklass is a useful and reliable toolkit for TCR repertoire analysis.
Related JoVE Video
Stable high-energy Q-switched resonantly diode-pumped Er:YAG laser at 1645??nm.
Appl Opt
PUBLISHED: 11-18-2014
Show Abstract
Hide Abstract
We demonstrate the generation of a stable high-energy Q-switched resonantly diode-pumped Er:YAG laser at 1645 nm in passive and active operation modes. For the passively Q-switched Er:YAG laser, the bi-layer graphene saturable absorber (SA), which was transferred onto the highly reflective cavity mirror by an improved method, was applied to deliver a stable pulse train with per-pulse energy of about 0.1 mJ. While in the active operation mode, the voltage-ON-type rubidium titanyl phosphate (RTP) Pockels cell as an electro-optic (EO) Q-switcher allows for the generation of a stable Q-switched pulse with per-pulse energy of up to 7.5 mJ. Our work may provide a basis for the development of a high-energy Er:YAG laser at 1645 nm to fulfill specific applications.
Related JoVE Video
Effects of Hyperbaric Oxygen on Symptoms and Quality of Life Among Service Members With Persistent Postconcussion Symptoms: A Randomized Clinical Trial.
JAMA Intern Med
PUBLISHED: 11-18-2014
Show Abstract
Hide Abstract
Improvement has been anecdotally observed in patients with persistent postconcussion symptoms (PCS) after mild traumatic brain injury following treatment with hyperbaric oxygen (HBO). The effectiveness of HBO as an adjunctive treatment for PCS is unknown to date.
Related JoVE Video
[The prevalence of risk factors and status of clinical practice patterns among hospitalized patients with acute coronary syndromes].
Zhonghua Nei Ke Za Zhi
PUBLISHED: 11-08-2014
Show Abstract
Hide Abstract
To analyze the distribution of multiple risk factors for hospitalized patients with acute coronary syndromes (ACS) and explore the status and determinants of drug usage recommended by the guideline.
Related JoVE Video
Pump-probe measurements of the thermal conductivity tensor for materials lacking in-plane symmetry.
Rev Sci Instrum
PUBLISHED: 11-03-2014
Show Abstract
Hide Abstract
We previously demonstrated an extension of time-domain thermoreflectance (TDTR) which utilizes offset pump and probe laser locations to measure in-plane thermal transport properties of multilayers. However, the technique was limited to systems of transversely isotropic materials studied using axisymmetric laser intensities. Here, we extend the mathematics so that data reduction can be performed on non-transversely isotropic systems. An analytic solution of the diffusion equation for an N-layer system is given, where each layer has a homogenous but otherwise arbitrary thermal conductivity tensor and the illuminating spots have arbitrary intensity profiles. As a demonstration, we use both TDTR and time-resolved magneto-optic Kerr effect measurements to obtain thermal conductivity tensor elements of <110> ?-SiO2. We show that the out-of-phase beam offset sweep has full-width half-maxima that contains nearly independent sensitivity to the in-plane thermal conductivity corresponding to the scanning direction. Also, we demonstrate a Nb-V alloy as a low thermal conductivity TDTR transducer layer that helps improve the accuracy of in-plane measurements.
Related JoVE Video
Modified Chimeric Radial Collateral Artery Perforator Flap for Repairing Hand Composite Defects.
J Reconstr Microsurg
PUBLISHED: 10-31-2014
Show Abstract
Hide Abstract
Background?The authors presented their clinical experience and demonstrated surgical methods for reconstructing complex bone and soft tissue defects of the hand by using modified chimeric radial collateral artery perforator flaps. Methods?Surgical procedures that employed 16 modified chimeric radial collateral artery perforator flaps and 3 dual paddle flaps were performed in 16 patients. Among the patients, eight had defects in the metacarpal bones and eight had defects in the phalanx bones. The flaps were created with a skin paddle and humeral bone segments by using independent perforators. The flaps ranged in size from 5.5?×?2.0 to 7.5?×?4.5 cm, whereas the humeral fragments ranged in size from 1.5?×?0.5 to 4.0?×?1.5 cm. The pedicle of the flaps was divided and ligated below the level at which the radial collateral artery separates into anterior and posterior branches. The recipient vessels were the proper digital artery, the palmar subcutaneous vein (n?=?12), the deep branch of the palmar ulnar artery (n?=?4), and the venae comitantes. The cosmetic appearance of both donor and recipient sites was evaluated during a follow-up visit. Results?Postoperative venous congestion occurred in two cases. The venous obstruction was reanastomosed after venous thrombectomy. The procedures were successful in both cases upon examination. All the flaps survived and all the donor sites were closed directly, leaving only a linear scar. Follow-up time ranged from 12 to 28 months. Bone components achieved union in all cases at an average of 5.4 months (ranging from 3 to 6 months). In two cases, the flap was defatted during the late postoperative period. Cosmetically acceptable results were achieved for the rest of the patients. Conclusion?The modified chimeric radial collateral artery perforator flap is a good alternative for reconstructing complex bone and soft tissue defects of the hands. Level of Evidence?This is a level IV, retrospective series.
Related JoVE Video
Robotic microsurgery: validating an assessment tool and plotting the learning curve.
Plast. Reconstr. Surg.
PUBLISHED: 10-31-2014
Show Abstract
Hide Abstract
The surgical robot has emerged as a potentially useful tool in microsurgery. The purpose of this study was to develop a validated assessment instrument and assess the learning curve for robotic microsurgery. The authors hypothesized that subjects would demonstrate improvement across multiple domains of performance with repetition of robotic microsurgical tasks.
Related JoVE Video
In Situ Transmission Electron Microscopy Probing of Native Oxide and Artificial Layers on Silicon Nanoparticles for Lithium Ion Batteries.
ACS Nano
PUBLISHED: 10-28-2014
Show Abstract
Hide Abstract
Surface modification of silicon nanoparticles via molecular layer deposition (MLD) has been recently proved to be an effective way for dramatically enhancing the cyclic performance in lithium ion batteries. However, the fundamental mechanism of how this thin layer of coating functions is not known, which is complicated by the inevitable presence of native oxide of several nanometers on the silicon nanoparticle. Using in situ TEM, we probed in detail the structural and chemical evolution of both uncoated and coated silicon particles upon cyclic lithiation/delithation. We discovered that upon initial lithiation, the native oxide layer converts to crystalline Li2O islands, which essentially increases the impedance on the particle, resulting in ineffective lithiation/delithiation and therefore low Coulombic efficiency. In contrast, the alucone MLD-coated particles show extremely fast, thorough, and highly reversible lithiation behaviors, which are clarified to be associated with the mechanical flexibility and fast Li(+)/e(-) conductivity of the alucone coating. Surprisingly, the alucone MLD coating process chemically changes the silicon surface, essentially removing the native oxide layer, and therefore mitigates side reactions and detrimental effects of the native oxide. This study provides a vivid picture of how the MLD coating works to enhance the Coulombic efficiency, preserves capacity, and clarifies the role of the native oxide on silicon nanoparticles during cyclic lithiation and delithiation. More broadly, this work also demonstrates that the effect of the subtle chemical modification of the surface during the coating process may be of equal importance to the coating layer itself.
Related JoVE Video
[Obstetric and neonatal outcomes of vanishing twin syndrome].
Nan Fang Yi Ke Da Xue Xue Bao
PUBLISHED: 10-28-2014
Show Abstract
Hide Abstract
To compare the obstetric and neonatal outcomes of singleton pregnancies originating from vanishing twin syndrome with those of singleton pregnancies originating from a single gestation following in vitro fertilization-embryo transfer (IVF-ET).
Related JoVE Video
Higher serum carotenoid concentrations associated with a lower prevalence of the metabolic syndrome in middle-aged and elderly Chinese adults.
Br. J. Nutr.
PUBLISHED: 10-28-2014
Show Abstract
Hide Abstract
The association between serum carotenoids and the metabolic syndrome (MetS) remains uncertain, and little is known about this relationship in the Chinese population. The present study examined the association between serum carotenoid concentrations and the MetS in Chinese adults. We conducted a community-based cross-sectional study in which 2148 subjects (1547 women and 601 men) aged 50-75 years were recruited in urban Guangzhou, China. Dietary data and other covariates were collected during face-to-face interviews. Blood pressure, waist circumference, blood lipids, glucose and serum carotenoids (?-, ?-carotene, ?-cryptoxanthin, lycopene and lutein/zeaxanthin) were examined. We found dose-response inverse relationships between individual serum carotenoid concentrations and total carotenoids and the prevalence of the MetS after adjusting for potential confounders (P for trend < 0·001). The OR of the MetS for the highest (v. lowest) quartile were 0·31 (95 % CI 0·20, 0·47) for ?-carotene, 0·23 (95 % CI 0·15, 0·36) for ?-carotene, 0·44 (95 % CI 0·29, 0·67) for ?-cryptoxanthin, 0·39 (95 % CI 0·26, 0·58) for lycopene, 0·28 (95 % CI 0·18, 0·44) for lutein+zeaxanthin and 0·19 (95 % CI 0·12, 0·30) for total carotenoids. Higher concentrations of each individual carotenoid and total carotenoids were significantly associated with a decrease in the number of abnormal MetS components (P for trend < 0·001-0·023). Higher serum carotenoid levels were associated with a lower prevalence of the MetS and fewer abnormal MetS components in middle-aged and elderly Chinese adults.
Related JoVE Video
Multiple Intracranial Metastatic Tumor Case Report and Aquaporin Water Channel-Related Research.
Cell Biochem. Biophys.
PUBLISHED: 10-25-2014
Show Abstract
Hide Abstract
This case report deals with multiple intracranial metastatic tumors and studies of expression and regulation characteristics of aquaporins (AQPs) of cerebellar metastatic tumor and brain tissue surrounding tumor. In this work, we try to understand the role of abnormal expression of AQPs in formation and elimination of brain edema and provide new ideas for the treatment of brain edema induced by tumor. The work involves resection of intracranial occupying lesions to get cerebellar metastatic tumor organization. Total RNA was extracted, RT-PCR was done, and immunohistochemical staining was done to study the expression and regulation characteristics of AQPs. We found that AQP4 had a high expression in the peritumoral brain tissue and no expression in the center of brain metastasis tumor organization. Around the tumor tissue, the AQP4 staining was junior in the more distant region from tumor and it added significantly in close to the tumor tissue region. It demonstrated that the AQP4 expression was upregulated, obviously with the distance drawing near gradually to tumor tissue. In addition, stained AQP1 was not observed on cerebellar metastatic tumor and peritumoral brain microvascular endothelial cells. The phenomenon that AQP4 had an increased expression in the surrounding region of cerebellar metastatic tumor and, moreover, increased significantly in the region next to the cerebellar metastatic tumor tightly is probably related to the formation of peritumoral brain edema and plays an important role in cytotoxic brain edema mechanism. AQP1 was not expressed on cerebellar metastatic tumor and peritumoral brain tissue microvascular endothelial cells, and this may be an important factor that the peritumoral interstitial brain edema is removed ineffectively to cause 'small tumor, big edema.'
Related JoVE Video
[The relevance of leukotriene D4 synthase gene A(-444)C polymorphism to clinical responsiveness of leukotriene receptor antagonist in patients with allergic rhinitis].
Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za Zhi
PUBLISHED: 10-22-2014
Show Abstract
Hide Abstract
To investigate the leukotriene D4 synthase gene A (LTD4S A)-444 C polymorphism in persistent allergic rhinitis (AR) of Chinese Han nationality and to evaluate its relevance to clinical responsiveness of leukotriene receptor antagonist.
Related JoVE Video
Cellular microRNAs up-regulate transcription via interaction with promoter TATA-box motifs.
RNA
PUBLISHED: 10-21-2014
Show Abstract
Hide Abstract
The TATA box represents one of the most prevalent core promoters where the pre-initiation complexes (PICs) for gene transcription are assembled. This assembly is crucial for transcription initiation and well regulated. Here we show that some cellular microRNAs (miRNAs) are associated with RNA polymerase II (Pol II) and TATA box-binding protein (TBP) in human peripheral blood mononuclear cells (PBMCs). Among them, let-7i sequence specifically binds to the TATA-box motif of interleukin-2 (IL-2) gene and elevates IL-2 mRNA and protein production in CD4(+) T-lymphocytes in vitro and in vivo. Through direct interaction with the TATA-box motif, let-7i facilitates the PIC assembly and transcription initiation of IL-2 promoter. Several other cellular miRNAs, such as mir-138, mir-92a or mir-181d, also enhance the promoter activities via binding to the TATA-box motifs of insulin, calcitonin or c-myc, respectively. In agreement with the finding that an HIV-1-encoded miRNA could enhance viral replication through targeting the viral promoter TATA-box motif, our data demonstrate that the interaction with core transcription machinery is a novel mechanism for miRNAs to regulate gene expression.
Related JoVE Video
Sub-10 nm Monoclinic Gd2O3:Eu(3+) Nanoparticles as Dual-Modal Nanoprobes for Magnetic Resonance and Fluorescence Imaging.
Langmuir
PUBLISHED: 10-20-2014
Show Abstract
Hide Abstract
Monoclinic Gd2O3:Eu(3+) nanoparticles (NPs) possess favorable magnetic and optical properties for biomedical application. However, how to obtain small enough NPs still remains a challenge. Here we combined the standard solid-state reaction with the laser ablation in liquids (LAL) technique to fabricate sub-10 nm monoclinic Gd2O3:Eu(3+) NPs and explained their formation mechanism. The obtained Gd2O3:Eu(3+) NPs exhibit bright red fluorescence emission and can be successfully used as fluorescence probe for cells imaging. In vitro and in vivo magnetic resonance imaging (MRI) studies show that the product can also serve as MRI good contrast agent. Then, we systematically investigated the nanotoxicity including cell viability, apoptosis in vitro, as well as the immunotoxicity and pharmacokinetics assays in vivo. This investigation provides a platform for the fabrication of ultrafine monoclinic Gd2O3:Eu(3+) NPs and evaluation of their efficiency and safety in preclinical application.
Related JoVE Video
Polymer optical fiber twisted macro-bend coupling system for liquid level detection.
Opt Express
PUBLISHED: 10-17-2014
Show Abstract
Hide Abstract
The liquid level detection principle of cladding mode frustrated total internal reflection (CMFTIR) effect is proposed. The significant enhancement of CMFTIR effect is realized through macro-bend coupling system in which the dark-field coupling phenomenon between two multimode polymer optic fibers is observed through experiment. Especially twisted macro-bend coupling structure (TMBCS) is adopted to achieve stable coupling of two naked POF. The testing result showed that the dark-filed forward coupling efficiency reached 2‰ and the extinction ratio of the liquid level probe reached 4.18dB. Compared with existing optical fiber liquid level sensors, the TMBCS probe is simpler, robuster, and cheaper. In addition, the TMBCS has the potential for displacement or stress sensing.
Related JoVE Video
Regulation of FSP27 protein stability by AMPK and HSC70.
Am. J. Physiol. Endocrinol. Metab.
PUBLISHED: 10-16-2014
Show Abstract
Hide Abstract
Fat specific protein 27 (FSP27) plays a pivotal role in controlling the formation of large lipid droplet and energy metabolism. The cellular levels of FSP27 are tightly regulated through the proteasomal ubiquitin-mediated degradation. However, the upstream signals that trigger FSP27 degradation and the underlying mechanism(s) have yet to be identified. Here we show that AMP-activated protein kinase (AMPK) activation by AICAR (5-amino-1-?-D-ribofuranosyl-imidazole-4-carboxamide) or phenformin induced the ubiquitination of FSP27 and promoted its degradation in 3T3-L1 adipocytes. The levels of FSP27 protein could be maintained by either knocking down AMPK?1 or blocking proteasomal pathway. Moreover, AICAR treatment induced multilocularization of LDs in 3T3-L1 adipocytes, reminiscent of the morphological changes in cells depleted of FSP27. Furthermore, mass spectrometry-based proteomic analysis identified heat shock cognate 70 (HSC70) as a novel binding protein of FSP27. The specific interaction was confirmed by co-immunoprecipitation of both ectopically expressed and endogenous proteins. Importantly, knockdown of HSC70 by small interference RNA resulted in increased half-life of FSP27 in cells treated with a protein synthesis inhibitor cycloheximide (CHX) or AICAR. However, silencing of the E3 ubiquitin ligase CHIP (C terminus of HSC70-Interacting Protein) failed to alter the stability of FSP27 protein under both conditions. Taken together, our data indicate that AMPK is a negative regulator of FSP27 stability through the proteasomal ubiquitin-dependent protein catabolic process. Promotion of FSP27 degradation may be an important factor responsible for the beneficial effect of AMPK activators on energy metabolism.
Related JoVE Video
Detailed simulation of the role of functionalized polymer chains on the structural, dynamic and mechanical properties of polymer nanocomposites.
Soft Matter
PUBLISHED: 10-09-2014
Show Abstract
Hide Abstract
To systematically study the effect of functionalized chain groups on polymer nanocomposites, we perform our simulation work in the following two ways. In the case of dilute loading of nanoparticles (NPs) with different geometries (spherical, sheet-like, rod-like NPs), we adopt coarse-grained molecular dynamics simulation to study the structural, dynamic and mechanical properties of polymer nanocomposites influenced by the terminal groups of linear polymer chains. We observe that the terminal groups have more probability to be adsorbed onto the surface of NPs with decreasing temperature, chain molecular weight and increasing chain stiffness. For all NPs with different geometries, more terminal groups segregate into the surface of NPs with increase in the interaction energy ?f-n between the terminal groups and the NPs. We also notice that the attractive interaction between the terminal groups and the sheet-like NPs induces the appearance of a gradient of translational dynamics of polymer chains, and the relaxation at the chain length scale is evidently different for various adsorbed layers, whereas the segmental relaxation only becomes slightly slower nearby the sheet-like NPs. For both pure and filled systems with spherical NPs, it is found that the stress-strain curves and bond orientations are significantly enhanced with increase in the interaction strength between the terminal groups as well as terminal groups and NPs. In the case of concentrated loading of NPs, we construct the atomistic models of C60, CNT and graphene to accurately account for the "many body effect." We explore the influence of the functionalization position along the chain backbone on the dispersion kinetics, realizing that the end-functionalization is more effective. The end-groups effect on the chain configuration, chain packing and graphene equilibrium dispersibility is examined. The translational and rotational (segmental and terminal relaxation) dynamics influenced by the interactions between the end groups and graphene are probed by tuning ?f-n and the volume fraction of graphene ?. Moreover, the shift in the glass transition temperature influenced by ?f-n and ? is quantitatively estimated by fitting the temperature dependence of the relaxation time using the Vogel-Fulcher-Tammann (VFT) equation. This work is hoped to provide a deep understanding of the polymer nanocomposites with functionalized polymer chains.
Related JoVE Video
[FLT3 mutation in acute promyelocytic leukemia patients with extramedullary relapse].
Sichuan Da Xue Xue Bao Yi Xue Ban
PUBLISHED: 10-08-2014
Show Abstract
Hide Abstract
To evaluate the role of FLT3 gene mutation in acute promyelocytic leukemia (APL) patients with extramedullary relapse.
Related JoVE Video
Facile synthesis of highly porous ni-sn intermetallic microcages with excellent electrochemical performance for lithium and sodium storage.
Nano Lett.
PUBLISHED: 10-08-2014
Show Abstract
Hide Abstract
Highly porous Ni3Sn2 microcages composed of tiny nanoparticles were synthesized by a facile template-free solvothermal method (based on Ostwald ripening and etching mechanism) for use as anode materials for high-capacity and high-rate-capability Li-ion and Na-ion batteries. The Ni3Sn2 porous microcages exhibit highly stable and substantial discharge capacities of the amount to 700 mA h g(-1) after 400 cycles at 0.2C and 530 mA h g(-1) after 1000 cycles at 1C for Li-ion battery anode. For Na-ions storage performance, a reversible capacity of approximate 270 mA h g(-1) is stably maintained at 1C during the first 300 cycles.
Related JoVE Video
Crazy paving pattern caused by pulmonary alveolar proteinosis: CT findings and the pathologic basis.
Zhong Nan Da Xue Xue Bao Yi Xue Ban
PUBLISHED: 10-02-2014
Show Abstract
Hide Abstract
To explore CT findings and pathologic basis of crazy paving pattern caused by pulmonary alveolar proteinosis.
Related JoVE Video
Disruption of the immune-checkpoint VISTA gene imparts a proinflammatory phenotype with predisposition to the development of autoimmunity.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 09-29-2014
Show Abstract
Hide Abstract
V domain-containing Ig suppressor of T-cell activation (VISTA) is a negative checkpoint regulator that suppresses T cell-mediated immune responses. Previous studies using a VISTA-neutralizing monoclonal antibody show that VISTA blockade enhances T-cell activation. The current study describes a comprehensive characterization of mice in which the gene for VISTA has been deleted. Despite the apparent normal hematopoietic development in young mice, VISTA genetic deficiency leads to a gradual accumulation of spontaneously activated T cells, accompanied by the production of a spectrum of inflammatory cytokines and chemokines. Enhanced T-cell responsiveness was also observed upon immunization with neoantigen. Despite the presence of multiorgan chronic inflammation, aged VISTA-deficient mice did not develop systemic or organ-specific autoimmune disease. Interbreeding of the VISTA-deficient mice with 2D2 T-cell receptor transgenic mice, which are predisposed to the development of experimental autoimmune encephalomyelitis, drastically enhanced disease incidence and intensity. Disease development is correlated with the increase in the activation of encephalitogenic T cells in the periphery and enhanced infiltration into the CNS. Taken together, our data suggest that VISTA is a negative checkpoint regulator whose loss of function lowers the threshold for T-cell activation, allowing for an enhanced proinflammatory phenotype and an increase in the frequency and intensity of autoimmunity under susceptible conditions.
Related JoVE Video
[A cone-beam CT study on morphology and position of condylar processes in skeletal Class III adult patients with mandibular deviation].
Zhonghua Kou Qiang Yi Xue Za Zhi
PUBLISHED: 09-27-2014
Show Abstract
Hide Abstract
To evaluate the morphology and position of condylar processes in skeletal Class III adult patients with mandibular deviation.
Related JoVE Video
[Reflection on the exfoliation of the tunic albuginea after the low-temperature plasma operation].
Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za Zhi
PUBLISHED: 09-25-2014
Show Abstract
Hide Abstract
To discuss the potential problems caused by the exfoliation of the tunic albuginea after the low-temperature plasma operation.
Related JoVE Video
Distinctive metabolite profiles in in-migrating sockeye salmon suggest sex-linked endocrine perturbation.
Environ. Sci. Technol.
PUBLISHED: 09-24-2014
Show Abstract
Hide Abstract
The health of Skeena River Sockeye salmon (Onchorhychus nerka) has been of increasing concern due to declining stock returns over the past decade. In the present work, in-migrating Sockeye from the 2008 run were evaluated using a mass spectrometry-based, targeted metabolomics platform. Our objectives were to (a) investigate natural changes in a subset of the hepatic metabolome arising from migration-associated changes in osmoregulation, locomotion, and gametogenesis, and (b) compare the resultant profiles with animals displaying altered hepatic vitellogenin A (vtg) expression at the spawning grounds, which was previously hypothesized as a marker of xenobiotic exposure. Of 203 metabolites monitored, 95 were consistently observed in Sockeye salmon livers and over half of these changed significantly during in-migration. Among the most dramatic changes in both sexes were a decrease in concentrations of taurine (a major organic osmolyte), carnitine (involved in fatty acid transport), and two major polyunsaturated fatty acids (eicosapentaenoic acid and docosahexaenoic acid). In females, an increase in amino acids was attributed to protein catabolism associated with vitellogenesis. Animals with atypical vtg mRNA expression demonstrated unusual hepatic amino acid, fatty acid, taurine, and carnitine profiles. The cause of these molecular perturbations remains unclear, but may include xenobiotic exposure, natural senescence, and/or interindividual variability. These data provide a benchmark for further investigation into the long-term health of migrating Skeena Sockeye.
Related JoVE Video
A comparative study of transulnar and transradial artery access for percutaneous coronary intervention in patients with acute coronary syndrome.
J Interv Cardiol
PUBLISHED: 09-22-2014
Show Abstract
Hide Abstract
Transradial access has become commonly used for elective evaluation of patients with coronary artery disease, but it has some disadvantages and has had limited use in the acute coronary syndrome (ACS). Because the diameter of the ulnar artery is usually larger than that of the radial artery, we hypothesized that the ulnar artery could be used as an access for percutaneous coronary intervention (PCI). The present study compares the feasibility, safety, and outcome of transulnar artery and transradial artery access for PCI in patients with ACS.
Related JoVE Video
[Effect of estradiol on cholesterol metabolism in J774a.1 mouse mononuclear/macrophage cells].
Yao Xue Xue Bao
PUBLISHED: 09-20-2014
Show Abstract
Hide Abstract
To explore the anti-atherosclerotic mechanism of estrogen and especially observe the effect of estradiol on the content of cholesterol in J774a.1 mouse mononuclear/macrophage-derived foam cells which were incubated with oxidized low-density lipoproteins (ox-LDL). J774a.1 mouse mononuclear/macrophages were incubated with ox-LDL or with both ox-LDL and estradiol (1, 0.1 or 0.01 micromol x L(-1)). Oil red O staining was used to observe the formation of foam cells, and cholesterol oxidase fluorometric was used to determine the content of cellular cholesterol content. Western blotting and RTFQ-PCR were used to observe the expressions of scavenger receptor class B type I (SR-B I ) in J774a.1 foam cells. Compared with the control cells, J774a.1 mouse mononuclear/macrophage-derived foam cells showed significantly increased contents of total cholesterol and cholesterol ester (P < 0.001) and decreased SR-B I mRNA expression (P < 0.01). Estradiol treatment significantly lowered the contents of total cholesterol and cholesterol ester (P < 0.05), and increased SR-B I protein and mRNA expression (P < 0.01) in the foam cells in a dose-dependent manner. Estradiol can inhibit the formation of mononuclear/macrophage-derived foam cells by decreasing the contents of total cholesterol and cholesterol ester and up-regulating the expression of SR-B I in the foam cells.
Related JoVE Video
Elevated expression of HMGA1 correlates with the malignant status and prognosis of non-small cell lung cancer.
Tumour Biol.
PUBLISHED: 09-18-2014
Show Abstract
Hide Abstract
High-mobility group A1 (HMGA1) has been suggested to play a significant role in tumor progression, but little is known about the accurate significance of HMGA1 in non-small cell lung cancer (NSCLC) patients. The aim of this study was to identify the role of HMGA1 in NSCLC. The expression status of HMGA1 was observed initially in NSCLC by Gene Expression Omnibus (GEO). The expression of HMGA1 messenger RNA (mRNA) and protein was examined in NSCLC and adjacent normal lung tissues through real-time PCR and immunohistochemistry. Meanwhile, the relationship of HMGA1 expression levels with clinical features and prognosis of NSCLC patients was analyzed. In our results, HMGA1 was overexpressed in NSCLC tissues compared with adjacent normal lung tissues in microarray data (GSE19804). HMGA1 mRNA and protein expressions were markedly higher in NSCLC tissues than in normal lung tissues (P?
Related JoVE Video
Concise synthesis of 2,4-disubstituted thiazoles from ?-azido disulfides and carboxylic acids or anhydrides: asymmetric synthesis of cystothiazole C.
Org. Biomol. Chem.
PUBLISHED: 09-17-2014
Show Abstract
Hide Abstract
A novel and efficient method for the one-pot synthesis of 2,4-disubstituted thiazoles from carboxylic acids or anhydrides is presented. Based on this new method, the total synthesis of the bis-2,4-disubstituted bis(thiazoles) natural product cystothiazole C is also presented.
Related JoVE Video
[Clinical literature based statistical analysis of common Chinese medical syndrome types].
Zhongguo Zhong Xi Yi Jie He Za Zhi
PUBLISHED: 09-17-2014
Show Abstract
Hide Abstract
According to the principle of evidence-based medicine (EBM), Chinese medical literatures based descriptive statistical analysis of common Chinese medical syndrome types were performed. By data extraction, standardization, and frequency calculation of disease names and syndrome types from 286 literatures in line with the inclusion criteria, the frequencies of diseases and syndromes were obtained to analyze common syndrome types in clinical practice, to analyze the distribution features of disease related syndromes and syndrome related diseases, to analyze the distribution of basic Chinese medical syndrome types in clinical common diseases as a whole, thus providing reference for clinical and basic researches.
Related JoVE Video
Knockdown of cancerous inhibitor of protein phosphatase 2A may sensitize metastatic castration-resistant prostate cancer cells to cabazitaxel chemotherapy.
Tumour Biol.
PUBLISHED: 09-12-2014
Show Abstract
Hide Abstract
Cancerous inhibitor of protein phosphatase 2A (CIP2A) is a recently identified human oncoprotein that can stabilize some proteins by inhibiting degradation mediated by protein phosphatase 2A (PP2A), and its level in cancer is associated with resistance to chemotherapy. However, whether CIP2A could increase chemoresistance of prostate cancer (PCa) cells to chemotherapeutic agent cabazitaxel remains unclear. To determine whether CIP2A serves as a potential therapeutic target of human PCa, we utilized small interference RNA (siRNA) to knock down CIP2A expression in human PCa cells and analyzed their phenotypic changes. The data demonstrated that CIP2A was significantly elevated in mCRPC cell lines C4-2 and ARCaPM at both the mRNA and protein levels. CIP2A silencing led to decreased proliferation and enhanced chemosensitivity and apoptosis to cabazitaxel in human PCa cells, as well as reduced Akt phosphorylation. Our data suggesting critical roles of CIP2A in PCa cells chemoresistance to cabazitaxel and raising the possibility of CIP2A inhibition as a promising approach for chemosensitization of metastatic castration-resistant prostate cancer (mCRPC).
Related JoVE Video
Wraparound Chimeric Radial Collateral Artery Perforator Flap for Reconstruction of Thumb Loss.
J Reconstr Microsurg
PUBLISHED: 09-11-2014
Show Abstract
Hide Abstract
Objective?We present our clinical experience and demonstrate surgical methods to reconstruct the thumb by using a wraparound chimeric radial collateral artery perforator flap. Methods?Surgical procedures were performed in 12 patients. Flaps with a skin paddle and humeral bone segment were created on the basis of independent perforators. The sizes of the flaps and humeral fragments ranged from 5.5 cm?×?2.0 cm to 7.5 cm?×?4.5 cm and from 1.5 cm?×?0.5 cm to 4.5 cm?×?1.5 cm, respectively. The flap pedicle was divided and ligated above the level at which the radial collateral artery was divided into anterior and posterior branches. The following recipient vessels were used: (1) the proper radial digital artery of the thumb and the palmar subcutaneous vein (n?=?8) and (2) the radial artery (n?=?4) and the venae comitantes. Nerve suture was conducted between the posterior cutaneous nerve of the arm and the proper ulnar digital nerve of the thumb. The cosmetic appearance of the donor and recipient sites and the static two-point discrimination of the operated finger were evaluated in a follow-up visit. Results?Postoperative venous congestion occurred in one case, but this complication was successfully treated after surgery. All of the flaps survived and all of the donor sites were closed directly, leaving a linear scar. Follow-up time ranged from 12 to 28 months. The union of bone components was observed in all of the cases at an average period of 4.5 months (range 3-6 months). Flap defatting was performed in two cases during the late postoperative period. Cosmetically acceptable results were achieved for the rest of the patients. The average of the static two-point discrimination scores was 9?mm (range 7-10 mm). Conclusions?The wraparound chimeric radial collateral artery perforator flap could be an effective option for thumb reconstruction because no major donor-site complications were found.
Related JoVE Video
A multicenter study of the clinical features of allergic rhinitis in central China.
Am J Rhinol Allergy
PUBLISHED: 09-09-2014
Show Abstract
Hide Abstract
Several epidemiological surveys of allergic rhinitis (AR) have been conducted in China. However, the clinical features of AR are still not clear enough. The aim of the current study was to perform a multicenter investigation to evaluate the clinical features of AR in China.
Related JoVE Video
Role of block copolymer morphology on particle percolation of polymer nanocomposites.
Soft Matter
PUBLISHED: 09-04-2014
Show Abstract
Hide Abstract
In this study, the effects of nanoparticle volume fraction, block stiffness, and diblock composition on the microstructure and electrical properties of composites are investigated using molecular dynamics simulation. It is shown that selective localization of conductive nanoparticles in a continuous block of diblock copolymer can dramatically reduce the percolation threshold. In the flexible-flexible copolymer systems with a relatively low particle loading, as the ratio of two blocks varies, one sees four kinds of phase structure: signal continuous, lamellar, co-continuous, and dispersed, corresponding to the order-disorder and continuity-dispersion transitions. In consideration of particle connectivity, the best electrical performance can be achieved with a special tri-continuous microstructure. While in the semi-flexible systems, the existence of rigid blocks can destroy the lamellar structure. If particles are located in the flexible block, a moderate stiffness of the rigid block can extensively enlarge the tri-continuous region, and high conductivity can be realized over a wide range of diblock compositions. If particles are located in the rigid block, however, high conductivity only emerges in a narrow composition range. In addition, the block should be prevented from becoming overstiff because this will cause direct particle aggregation.
Related JoVE Video
Characterization and plasmid elimination of NDM-1-producing Acinetobacter calcoaceticus from China.
PLoS ONE
PUBLISHED: 09-02-2014
Show Abstract
Hide Abstract
The presence of multidrug-resistant bacterial pathogens in the environment poses a serious threat to public health. The opportunistic Acinetobacter spp. are among the most prevalent causes of nosocomial infections. Here, we performed complete genome sequencing of the Acinetobacter calcoaceticus strain XM1570, which was originally cultivated from the sputum of a patient diagnosed with pneumonia in Xiamen in 2010. We identified carbapenem resistance associated gene bla(NDM-1) located on a 47.3-kb plasmid. Three methods--natural reproduction, sodium dodecyl sulfate treatment and nalidixic acid treatment--were used to eliminate the bla(NDM-1)-encoding plasmid, which achieved elimination rates of 3.32% (10/301), 83.78% (278/332), and 84.17% (298/354), respectively. Plasmid elimination dramatically increased antibiotic sensitivity, reducing the minimum bacteriostatic concentration of meropenem from 256 µg/ml in the clinical strain to 0.125 µg/ml in the plasmid-eliminated strain. Conjugation transfer assays showed that the bla(NDM-1)-containing plasmid could be transferred into Escherichia coli DH5?:pBR322 in vitro as well as in vivo in mice. The bla(NDM-1) genetic environment was in accordance with that of other bla(NDM-1) genes identified from India, Japan, and Hong-Kong. The multilocus sequence type of the isolate was identified as ST-70. Two novel genes encoding intrinsic OXA and ADC were identified and named as OXA-417 and ADC-72. The finding of bla(NDM-1) in species like A. calcoaceticus demonstrates the wide spread of this gene in gram-negative bacteria which is possible by conjugative plasmid transfer. The results of this study may help in the development of a treatment strategy for controlling NDM-1 bacterial infection and transmission.
Related JoVE Video
Clostridium beijerinckii mutant obtained atmospheric pressure glow discharge generates enhanced electricity in a microbial fuel cell.
Biotechnol. Lett.
PUBLISHED: 09-02-2014
Show Abstract
Hide Abstract
A Clostridium beijerinckii mutant M13 was derived from C. beijerinckii NCIMB 8052 by atmospheric pressure glow discharge. C. beijerinckii M13 generated a maximum output power density of 79.2 mW m(-2) and a maximum output voltage of 230 mV in a microbial fuel cell containing 1 g glucose l(-1) as carbon source and 0.15 g methyl viologen l(-1) as an electron carrier.
Related JoVE Video
Influence of various nanoparticle shapes on the interfacial chain mobility: a molecular dynamics simulation.
Phys Chem Chem Phys
PUBLISHED: 09-02-2014
Show Abstract
Hide Abstract
To fully understand the polymer-filler interfacial interaction mechanism, we use a coarse-grained molecular dynamics simulation to mainly investigate the interfacial dynamic properties by tuning the polymer-filler interaction, temperature, chain length, volume fraction of filler, and size and shape of filler. The polymer beads around the filler exhibit an obvious layering behavior and a gradient of polymer dynamics is observed for systems filled with three kinds of fillers (spherical, rod-like and sheet-like). By analyzing the dynamics of the interfacial beads in the first adsorbed layer, we find that the mobility of interfacial beads becomes greater at lower polymer-filler interaction strength and higher temperature. The adsorption/desorption dynamics of interfacial beads decreases with the increase of the chain length, and then becomes nearly unchanged when the chain length exceeds the entanglement molecular weight. It is found that the influence of the different size of nanospheres on the mobility of interfacial beads is just induced by the curvature. However, for systems filled with nanorods and nanosheets, besides the curvature, the force exerted on the polymer beads also plays an important role. For systems filled with three kinds of fillers, the mobility of interfacial beads is the slowest for the nanosheet filled system and only in this case the "glassy layer" exists for strongly attractive interfacial interaction. By comparing the dynamics of the adsorbed polymer beads for three different shapes of filler, it is concluded that it is the total force exerted on the polymer beads by the filler that determines the mobility of the interfacial beads. In short, this work could provide valuable information on the fundamental understanding of polymer-filler interfacial behavior, especially for systems filled with fillers of different shapes.
Related JoVE Video
Synthesis and anticancer evaluation of benzyloxyurea derivatives.
Chem. Pharm. Bull.
PUBLISHED: 09-02-2014
Show Abstract
Hide Abstract
A series of novel benzyloxyurea derivatives was designed, synthesized by substituting different benzyls or phenyls on N,N'-positions of the hydroxyurea (HU). These target compounds were evaluated for their anticancer activity in vitro against human leukemia cell line K562 and murine leukemia cell line L1210 in comparison with HU by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Some of the compounds showed promising anticancer activity against the cells. Molecular docking experiments with Saccharomyces cerevisiae R1 domain indicated that 4a and 4f' have stronger affinity than 4m and 4n. Flow cytometry study showed that compound 4g exerted greater apoptotic activity against K562 cells line than HU.
Related JoVE Video
[Transient hepatic venous occlusion induced liver hemodynamic change and reperfusion injury in rats].
Zhongguo Yi Xue Ke Xue Yuan Xue Bao
PUBLISHED: 09-02-2014
Show Abstract
Hide Abstract
To observe the hemodynamic change and reperfusion injury cause by transient hepatic venous occlusion and transient hepatic inflow occlusion in rats.
Related JoVE Video
A three-gene panel that distinguishes benign from malignant thyroid nodules.
Int. J. Cancer
PUBLISHED: 08-30-2014
Show Abstract
Hide Abstract
Reliable preoperative diagnosis of malignant thyroid tumors remains challenging because of the inconclusive cytological examination of fine-needle aspiration biopsies. Although numerous studies have successfully demonstrated the use of high-throughput molecular diagnostics in cancer prediction, the application of microarrays in routine clinical use remains limited. Our aim was, therefore, to identify a small subset of genes to develop a practical and inexpensive diagnostic tool for clinical use. We developed a two-step feature selection method composed of a linear models for microarray data (LIMMA) linear model and an iterative Bayesian model averaging model to identify a suitable gene set signature. Using one public dataset for training, we discovered a three-gene signature dipeptidyl-peptidase 4 (DPP4), secretogranin V (SCG5) and carbonic anhydrase XII (CA12). We then evaluated the robustness of our gene set using three other independent public datasets. The gene signature accuracy was 85.7, 78.8 and 85.7%, respectively. For experimental validation, we collected 70 thyroid samples from surgery and our three-gene signature method achieved an accuracy of 94.3% by quantitative polymerase chain reaction (QPCR) experiment. Furthermore, immunohistochemistry in 29 samples showed proteins expressed by these three genes are also differentially expressed in thyroid samples. Our protocol discovered a robust three-gene signature that can distinguish benign from malignant thyroid tumors, which will have daily clinical application.
Related JoVE Video
Morphological Features of the Neonatal Brain Following Exposure to Regional Anesthesia During Labor and Delivery.
Magn Reson Imaging
PUBLISHED: 08-29-2014
Show Abstract
Hide Abstract
Recent animal and human epidemiological studies suggest that early childhood exposure to anesthesia may have adverse effects on brain development. As more than 50% of pregnant women in the United States and one-third in the United Kingdom receive regional anesthesia during labor and delivery, understanding the effects of perinatal anesthesia on postnatal brain development has important public health relevance.
Related JoVE Video
Effect of Individual Fc Methionine Oxidation on FcRn Binding: Met252 Oxidation Impairs FcRn Binding More Profoundly than Met428 Oxidation.
J Pharm Sci
PUBLISHED: 08-29-2014
Show Abstract
Hide Abstract
The long serum half-lives of mAbs are conferred by pH-dependent binding of IgG-Fc to the neonatal Fc receptor (FcRn). The Fc region of human IgG1 has three conserved methionine residues, Met252, Met358, and Met428. Recent studies showed oxidation of these Met residues impairs FcRn binding and consequently affects pharmacokinetics of therapeutic antibodies. However, the quantitative effect of individual Met oxidation on Fc-FcRn binding has not been addressed. This information is valuable for defining critical quality attributes. In the present study, two sets of homodimeric site-directed IgG1 mutations were generated to understand how individual Fc Met oxidation affects FcRn binding. The first approach used Met to Leu mutants to block site-specific Met oxidation. In the other approach, Met to Gln mutants were designed to mimic site-specific Met oxidation. Both mutagenesis approaches show that either Met252 or Met428 oxidation alone significantly impairs Fc-FcRn binding. Met252 oxidation has a more deleterious effect on FcRn binding than M428 oxidation, whereas Met428 oxidation has a bigger destabilization effect on the thermal stability. Our results also show that Met358 oxidation does not affect FcRn binding. In addition, our study suggests that Met to Gln mutation may serve as an important tool to understand Met oxidation. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci.
Related JoVE Video
?-aminobutyric acid receptors affect the progression and migration of tumor cells.
J. Recept. Signal Transduct. Res.
PUBLISHED: 08-28-2014
Show Abstract
Hide Abstract
Abstract ?-aminobutyric acid (GABA) is a multifunctional molecule found in the nervous system and non-neuronal tissues. GABA receptors combine with GABA molecules and transmit signal stimuli into cells. In addition to traditional neurotransmission and regulation of secretion, GABA and GABA receptors are involved in cell differentiation and proliferation throughout peripheral organs, as well as in tumorigenesis. The exact mechanism of the GABAergic system in regulating tumor development is unclear, but many studies have revealed that GABA receptors exert critical regulative effects on tumor cell proliferation and migration. In this review, the molecular structure, distribution and biological function of GABA receptors associated with tumorigenesis are described. Recent advances in the elucidation of mechanisms underlying GABAergic signaling control over tumor growth are also discussed.
Related JoVE Video
Airway mechanosensor behavior during application of positive end-expiratory pressure.
Respiration
PUBLISHED: 08-26-2014
Show Abstract
Hide Abstract
Positive end-expiratory pressure (PEEP) is commonly used in clinical settings. It is expected to affect the input from slowly adapting pulmonary stretch receptors (SARs), leading to altered cardiopulmonary functions. However, we know little about how SARs behave during PEEP application.
Related JoVE Video
Comparative Effectiveness of Di'ao Xin Xue Kang Capsule and Compound Danshen Tablet in Patients With Symptomatic Chronic Stable Angina.
Sci Rep
PUBLISHED: 08-18-2014
Show Abstract
Hide Abstract
A high proportion of patients with stable angina remains symptomatic despite multiple treatment options. Di'ao Xinxuekang (XXK) capsule and Compound Danshen (CDS) tablet have been approved for treating angina pectoris for more than 20 years in China. We compare the anti-anginal effectiveness of XXK capsule and CDS tablet in patients with symptomatic chronic stable angina. A randomized, multicenter, double-blind, parallel-group, superiority trial was conducted in 4 study sites. 733 patients with symptomatic chronic stable angina were included in the full analysis set. The primary outcomes were the proportion of patients who were angina-free and the proportion of patients with normal electrocardiogram (ECG) recordings during 20 weeks treatment. Compared with CDS, XXK significantly increased the proportion of angina-free patients, but no significant difference was noted in the proportion of patients with normal ECG recordings. Weekly angina frequency and nitroglycerin use were significantly reduced with XXK versus CDS at week 20. Moreover, XXK also improved the quality of life of angina patients as measured by the SAQ score and Xueyu Zheng (a type of TCM syndrome) score. We demonstrate that XXK capsule is more effective for attenuating anginal symptoms and improving quality of life in patients with symptomatic chronic stable angina, compared with CDS tablet.
Related JoVE Video
Energy Stress Regulates Hippo-YAP Signaling Involving AMPK-Mediated Regulation of Angiomotin-like 1 Protein.
Cell Rep
PUBLISHED: 08-18-2014
Show Abstract
Hide Abstract
Hippo signaling is a tumor-suppressor pathway involved in organ size control and tumorigenesis through the inhibition of YAP and TAZ. Here, we show that energy stress induces YAP cytoplasmic retention and S127 phosphorylation and inhibits YAP transcriptional activity and YAP-dependent transformation. These effects require the central metabolic sensor AMP-activated protein kinase (AMPK) and the upstream Hippo pathway components Lats1/Lats2 and angiomotin-like 1 (AMOTL1). Furthermore, we show that AMPK directly phosphorylates S793 of AMOTL1. AMPK activation stabilizes and increases AMOTL1 steady-state protein levels, contributing to YAP inhibition. The phosphorylation-deficient S793Ala mutant of AMOTL1 showed a shorter half-life and conferred resistance to energy-stress-induced YAP inhibition. Our findings link energy sensing to the Hippo-YAP pathway and suggest that YAP may integrate spatial (contact inhibition), mechanical, and metabolic signals to control cellular proliferation and survival.
Related JoVE Video
Prognostic significance of Ki-67 antigen expression in extranodal natural killer/T-cell lymphoma, nasal type.
Med. Oncol.
PUBLISHED: 08-18-2014
Show Abstract
Hide Abstract
Extranodal natural killer (NK)/T-cell lymphoma, nasal-type (ENKTL) is a clinically heterogeneous disease with poor prognosis and requires risk stratification in affected patients. Recent studies have shown that Ki-67 may serve as a prognostic marker in certain types of lymphoma. We analyzed Ki-67 expression and its correlation with prognosis in 182 patients with ENKTL from January 2002 to June 2013. The patients were classified into two groups through a median value: low (<60%) versus high Ki-67 (?60%). High Ki-67 expression was more common in patients with B symptoms (p=0.02), bulky disease (p=0.001), and extraupper aerodigestive tract NK/T-cell lymphoma (p=0.001). High Ki-67 expression was significantly associated with poor overall survival (p<0.0001) and progression-free survival (p<0.0001). For patients with low-risk IPI or KPI, Ki-67 at diagnosis could contribute to distinguish patients with favorable outcomes from those with poor outcomes. The results of multivariate analysis showed that the high Ki-67 expression is an independent prognostic factor for overall survival and progression-free survival. (OS, p=0.001; PFS, p=0.003). Our data showed that Ki-67 is an effective prognostic indicator of survival in ENKTL patients. This prognostic index may be helpful in identifying high-risk patients with ENKTL.
Related JoVE Video
Experimental study of needle-tissue interaction forces: effect of needle geometries, insertion methods and tissue characteristics.
J Biomech
PUBLISHED: 08-15-2014
Show Abstract
Hide Abstract
A thorough understanding of needle-tissue interaction mechanics is necessary to optimize needle design, achieve robotically needle steering, and establish surgical simulation system. It is obvious that the interaction is influenced by numerous variable parameters, which are divided into three categories: needle geometries, insertion methods, and tissue characteristics. A series of experiments are performed to explore the effect of influence factors (material samples n=5 for each factor) on the insertion force. Data were collected from different biological tissues and a special tissue-equivalent phantom with similar mechanical properties, using a 1-DOF mechanical testing system instrumented with a 6-DOF force/torque (F/T) sensor. The experimental results indicate that three basic phases (deformation, insertion, and extraction phase) are existent during needle penetration. Needle diameter (0.7-3.2mm), needle tip (blunt, diamond, conical, and beveled) and bevel angle (10-85°) are turned out to have a great influence on insertion force, so do the insertion velocity (0.5-10mm/s), drive mode (robot-assisted and hand-held), and the insertion process (interrupted and continuous). Different tissues such as skin, muscle, fat, liver capsule and vessel are proved to generate various force cures, which can contribute to the judgement of the needle position and provide efficient insertion strategy.
Related JoVE Video
Antioxidant and protective effect of inulin and catechin grafted inulin against CCl4-induced liver injury.
Int. J. Biol. Macromol.
PUBLISHED: 08-14-2014
Show Abstract
Hide Abstract
In this study, the antioxidant activity and hepatoprotective effect of inulin and catechin grafted inulin (catechin-g-inulin) against carbon tetrachloride (CCl4)-induced acute liver injury were investigated. Results showed that both inulin and catechin-g-inulin had moderate scavenging activity on superoxide radical, hydroxyl radical and H2O2, as well as lipid peroxidation inhibition effect. The antioxidant activity decreased in the order of Vc>catechin>catechin-g-inulin>inulin. Administration of inulin and catechin-g-inulin could significantly reduce the elevated levels of serum aspartate transaminase, alanine transaminase and alkaline phosphatase as compared to CCl4 treatment group. Moreover, inulin and catechin-g-inulin significantly increased the levels of hepatic superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione and total antioxidant capacity, whereas markedly decreased the malondialdehyde level when compared with CCl4 treatment group. Notably, catechin-g-inulin showed higher hepatoprotective effect than inulin. In addition, the hepatoprotective effect of catechin-g-inulin was comparable to positive standard of silymarin. Our results suggested that catechin-g-inulin had potent antioxidant activity and potential protective effect against CCl4-induced acute liver injury.
Related JoVE Video
The 18?kDa Translocator Protein, Microglia and Neuroinflammation.
Brain Pathol.
PUBLISHED: 08-13-2014
Show Abstract
Hide Abstract
The 18?kDa translocator protein (TSPO), previously known as the peripheral benzodiazepine receptor, is expressed in the injured brain. It has become known as an imaging marker of "neuroinflammation" indicating active disease, and is best interpreted as a nondiagnostic biomarker and disease staging tool that refers to histopathology rather than disease etiology. The therapeutic potential of TSPO as a drug target is mostly based on the understanding that it is an outer mitochondrial membrane protein required for the translocation of cholesterol, which thus regulates the rate of steroid synthesis. This pivotal role together with the evolutionary conservation of TSPO has underpinned the belief that any loss or mutation of TSPO should be associated with significant physiological deficits or be outright incompatible with life. However, against prediction, full Tspo knockout mice are viable and across their lifespan do not show the phenotype expected if cholesterol transport and steroid synthesis were significantly impaired. Thus, the "translocation" function of TSPO remains to be better substantiated. Here, we discuss the literature before and after the introduction of the new nomenclature for TSPO and review some of the newer findings. In light of the controversy surrounding the function of TSPO, we emphasize the continued importance of identifying compounds with confirmed selectivity and suggest that TSPO expression is analyzed within specific disease contexts rather than merely equated with the reified concept of "neuroinflammation."
Related JoVE Video
Farnesoid X receptor inhibits LNcaP cell proliferation via the upregulation of PTEN.
Exp Ther Med
PUBLISHED: 08-11-2014
Show Abstract
Hide Abstract
Prostate cancer is a form of cancer that develops in the prostate, a gland in the male reproductive system. In the present study, the activation of the farnesoid X receptor (FXR), a member of the nuclear receptor superfamily, was demonstrated to inhibit cell proliferation in LNcaP cells. Using clinical samples, mRNA and protein levels of FXR were found to be significantly decreased by quantitative PCR and western blot analysis in prostate cancer tissues. In vitro studies identified further that activation or overexpression of FXR suppressed prostate cancer cell proliferation as measured by BrdU incorporation assays. At the molecular level, the results further revealed that the expression of the tumor suppressor gene, PTEN, was upregulated by FXR activation. Therefore, the observations indicated that FXR functions as a tumor suppressor in prostate cancer, which may provide a novel method for molecular targeting cancer treatment.
Related JoVE Video
Restoration and protection of brachial plexus injury: hot topics in the last decade.
Neural Regen Res
PUBLISHED: 08-10-2014
Show Abstract
Hide Abstract
Brachial plexus injury is frequently induced by injuries, accidents or birth trauma. Upper limb function may be partially or totally lost after injury, or left permanently disabled. With the development of various medical technologies, different types of interventions are used, but their effectiveness is wide ranging. Many repair methods have phasic characteristics, i.e., repairs are done in different phases. This study explored research progress and hot topic methods for protection after brachial plexus injury, by analyzing 1,797 articles concerning the repair of brachial plexus injuries, published between 2004 and 2013 and indexed by the Science Citation Index database. Results revealed that there are many methods used to repair brachial plexus injury, and their effects are varied. Intervention methods include nerve transfer surgery, electrical stimulation, cell transplantation, neurotrophic factor therapy and drug treatment. Therapeutic methods in this field change according to the hot topic of research.
Related JoVE Video
Non-intubated complete thoracoscopic bronchial sleeve resection for central lung cancer.
J Thorac Dis
PUBLISHED: 08-09-2014
Show Abstract
Hide Abstract
Bronchial sleeve resection has emerged as an effective thoracoscopic approach for central lung cancer with reduced operation mortality rates, optimal lung function and long-term survival. Endobronchial intubation is a commonly used method of anesthesia for such thoracoscopic procedures, but is associated with increased intubation-related and lung complications. Non-intubated epidural anesthesia represents an alternative approach which may avoid such difficulties, particularly in complicated sleeve resection situations. Here we have described a case of complete endoscopic bronchial sleeve resection of right lower lung cancer under non-intubated epidural anesthesia.
Related JoVE Video
Expression of monellin in a food-grade delivery system in Saccharomyces cerevisiae.
J. Sci. Food Agric.
PUBLISHED: 08-06-2014
Show Abstract
Hide Abstract
Genetically modified (GM) foods have caused many controversies. Construction of a food-grade delivery system is desirable technique with presumptive impact on industrial applications from the perspective of bio-safety. The aim of this study was to construct a food-grade delivery system for Saccharomyces cerevisiae. Meanwhile monellin from the berries of the West African forest plant Dioscoreophyllum cumminsii was expressed in this system.
Related JoVE Video
Polycomb- and REST-associated histone deacetylases are independent pathways toward a mature neuronal phenotype.
Elife
PUBLISHED: 08-02-2014
Show Abstract
Hide Abstract
The bivalent hypothesis posits that genes encoding developmental regulators required for early lineage decisions are poised in stem/progenitor cells by the balance between a repressor histone modification (H3K27me3), mediated by the Polycomb Repressor Complex 2 (PRC2), and an activator modification (H3K4me3). In this study, we test whether this mechanism applies equally to genes that are not required until terminal differentiation. We focus on the RE1 Silencing Transcription Factor (REST) because it is expressed highly in stem cells and is an established global repressor of terminal neuronal genes. Elucidation of the REST complex, and comparison of chromatin marks and gene expression levels in control and REST-deficient stem cells, shows that REST target genes are poised by a mechanism independent of Polycomb, even at promoters which bear the H3K27me3 mark. Specifically, genes under REST control are actively repressed in stem cells by a balance of the H3K4me3 mark and a repressor complex that relies on histone deacetylase activity. Thus, chromatin distinctions between pro-neural and terminal neuronal genes are established at the embryonic stem cell stage by two parallel, but distinct, repressor pathways.
Related JoVE Video
Increased expressions of SATB1 and S100A4 are associated with poor prognosis in human colorectal carcinoma.
APMIS
PUBLISHED: 07-22-2014
Show Abstract
Hide Abstract
This study was designed to explore the correlation between expressions of SATB1 and S100A4 and their relationships to the clinicopathologic parameters of colorectal carcinoma (CRC). Expressions of SATB1 and S100A4 were evaluated by immunohistochemistry in a cohort of 131 primary CRC patients undergone surgical resection from 2005 to 2007. SATB1 and S100A4 were positively expressed in 48.9% and 54.2% of CRC cases, respectively. SATB1 and S100A4 expressions in tumor tissues were significantly higher than those in the corresponding normal tissues. A positive correlation was observed between SATB1 and S100A4. Moreover, the levels of SATB1 and S100A4 were both significantly associated with invasion, lymph node status, and TNM stage of CRC, whereas S100A4 expression was also correlated with distant metastasis. Multivariate analysis revealed that SATB1 expression was an independent prognostic indicator for poor survival of CRC. Further survival analysis indicated that co-expression of SATB1 and S100A4 suggested a worse 5-year overall survival rate in CRC patients. Thus, combined analysis of SATB1 and S100A4 expressions may be valuable in determining the development and progression of CRC. Co-expression of SATB1 and S100A4 is an unfavorable prognostic indicator and may be useful in the follow-up of patients with CRC.
Related JoVE Video
OsCERK1 and OsRLCK176 play important roles in peptidoglycan and chitin signaling in rice innate immunity.
Plant J.
PUBLISHED: 07-18-2014
Show Abstract
Hide Abstract
Microbe-associated molecular pattern (MAMP)-triggered immunity plays critical roles in the basal resistance defense response in plants. Chitin and peptidoglycan (PGN) are major molecular patterns for fungi and bacteria, respectively. Two rice (Oryza sativa) lysin motif-containing proteins, OsLYP4 and OsLYP6, function as receptors that sense bacterial PGN and fungal chitin. These membrane receptors, which lack intracellular kinase domains, likely contain another component for transmembrane immune signal transduction. Here, we demonstrate that the rice LysM receptor-like kinase OsCERK1, a key component of the chitin elicitor signaling pathway, also plays an important role in PGN-triggered immunity in rice. Silencing of OsCERK1 suppressed PGN-induced (and chitin-induced) immunity responses, including reactive oxygen species generation, defense gene expression, and callose deposition, indicating that OsCERK1 is essential for both PGN and chitin signaling initiated by OsLYP4 and OsLYP6. OsLYP4 associated with OsLYP6 and the rice chitin receptor CEBiP in the absence of PGN or chitin, and treatment with PGN or chitin led to their disassociation in vivo. OsCERK1 associated with OsLYP4 or OsLYP6 when induced by PGN but it associated with OsLYP4, OsLYP6, or CEBiP under chitin treatment, suggesting the presence of different patterns of ligand-induced heterooligomeric receptor complexes. Furthermore, the receptor-like cytoplasmic kinase OsRLCK176 functions downstream of OsCERK1 in the PGN and chitin signaling pathways, suggesting that these MAMPs share overlapping intracellular signaling components. Therefore, OsCERK1 plays dual roles in PGN and chitin signaling in rice innate immunity and as an adaptor involved in signal transduction at the plasma membrane in conjunction with OsLYP4 and OsLYP6. This article is protected by copyright. All rights reserved.
Related JoVE Video
microRNA-181 promotes prostate cancer cell proliferation by regulating DAX-1 expression.
Exp Ther Med
PUBLISHED: 07-16-2014
Show Abstract
Hide Abstract
microRNAs (miRNAs) are a class of short noncoding RNA molecules that have a critical role in the initiation and progression of types of human cancer, including prostate cancer. In the present study, the expression of miR-181 in prostate cancer tissues was evaluated and was demonstrated to be significantly upregulated in prostate cancer tissues compared with that in adjacent normal tissues. The results of in vitro MTT and BrdU incorporation assays, as well as cell-cycle analysis, indicated that miR-181 overexpression markedly promoted the proliferation of LNCaP cells. Furthermore, miR-181 overexpression was found to promote the progression of LNCaP tumor growth in nude mice. Mechanistic studies demonstrated that dosage-sensitive sex reversal, adrenal hypoplasia critical region, on chromosome X, gene 1 (DAX-1), a negative regulator of androgen receptor in prostate cancer, was inhibited by miR-181 overexpression. Therefore, the results from the present study suggest that miR-181 functions as a growth-suppressive miRNA during prostate cancer development.
Related JoVE Video
Next generation sequencing unravels the biosynthetic ability of Spearmint ( Mentha spicata ) peltate glandular trichomes through comparative transcriptomics.
BMC Plant Biol.
PUBLISHED: 07-09-2014
Show Abstract
Hide Abstract
BackgroundPlant glandular trichomes are chemical factories with specialized metabolic capabilities to produce diverse compounds. Aromatic mint plants produce valuable essential oil in specialised glandular trichomes known as peltate glandular trichomes (PGT). Here, we performed next generation transcriptome sequencing of different tissues of Mentha spicata (spearmint) to identify differentially expressed transcripts specific to PGT. Our results provide a comprehensive overview of PGT¿s dynamic metabolic activities which will help towards pathway engineering.ResultsSpearmint RNAs from 3 different tissues: PGT, leaf and leaf stripped of PGTs (leaf-PGT) were sequenced by Illumina paired end sequencing. The sequences were assembled de novo into 40,587 non-redundant unigenes; spanning a total of 101 Mb. Functions could be assigned to 27,025 (67%) unigenes and among these 3,919 unigenes were differentially expressed in PGT relative to leaf - PGT. Lack of photosynthetic transcripts in PGT transcriptome indicated the high levels of purity of isolated PGT, as mint PGT are non-photosynthetic. A significant number of these unigenes remained unannotated or encoded hypothetical proteins. We found 16 terpene synthases (TPS), 18 cytochrome P450s, 5 lipid transfer proteins and several transcription factors that were preferentially expressed in PGT. Among the 16 TPSs, two were characterized biochemically and found to be sesquiterpene synthases.ConclusionsThe extensive transcriptome data set renders a complete description of genes differentially expressed in spearmint PGT. This will facilitate the metabolic engineering of mint terpene pathway to increase yield and also enable the development of strategies for sustainable production of novel or altered valuable compounds in mint.
Related JoVE Video
Safety and feasibility of video-assisted thoracoscopic surgery for stage IIIA lung cancer.
Chin. J. Cancer Res.
PUBLISHED: 07-05-2014
Show Abstract
Hide Abstract
The current study was prospectively designed to explore the application of video-assisted thoracoscopic surgery (VATS) radical treatment for patients with stage IIIA lung cancer, with the primary endpoints being the safety and feasibility of this operation and the second endpoints being the survival and complications after the surgery.
Related JoVE Video
[Effects of salt stress on Nitraria roborowskii growth and physiological characteristics of stress resistance].
Ying Yong Sheng Tai Xue Bao
PUBLISHED: 07-03-2014
Show Abstract
Hide Abstract
A pot experiment was conducted to study the influences of different NaCl concentrations (0, 50, 100, 200, 400 mmol x L(-1)) on the growth, leaf hydrogen peroxide (H2O2), malonaldehyde (MDA) content, superoxide dismutases (SOD), catalases (CAT), peroxidase (POD), ascorbate peroxidase (APX) enzymes activity, water potential, soluble sugar and proline contents of Nitraria roborowskii. The results showed that the growth of N. roborowskii was not affected at lower NaCl concentrations (< or = 50 mmol L(-1)), while the SOD, POD, CAT and APX activities in leaves of N. roborowskii were increased. However, higher NaCl concentrations (> 50 mmol x L(-1)) restrained the growth parameters of crown area, number of branches, dry mass of leaf, branch and lateral root, and remarkably reduced the SOD, POD and CAT activities, soluble sugar and proline contents in leaves of N. roborowskii. H2O2 and MDA contents in leaves were increased and water potential was reduced with increasing NaCl concentrations.
Related JoVE Video
Derivation and expansion of PAX7-positive muscle progenitors from human and mouse embryonic stem cells.
Stem Cell Reports
PUBLISHED: 06-30-2014
Show Abstract
Hide Abstract
Cell therapies treating pathological muscle atrophy or damage requires an adequate quantity of muscle progenitor cells (MPCs) not currently attainable from adult donors. Here, we generate cultures of approximately 90% skeletal myogenic cells by treating human embryonic stem cells (ESCs) with the GSK3 inhibitor CHIR99021 followed by FGF2 and N2 supplements. Gene expression analysis identified progressive expression of mesoderm, somite, dermomyotome, and myotome markers, following patterns of embryonic myogenesis. CHIR99021 enhanced transcript levels of the pan-mesoderm gene T and paraxial-mesoderm genes MSGN1 and TBX6; immunofluorescence confirmed that 91% ± 6% of cells expressed T immediately following treatment. By 7 weeks, 47% ± 3% of cells were MYH(+ve) myocytes/myotubes surrounded by a 43% ± 4% population of PAX7(+ve) MPCs, indicating 90% of cells had achieved myogenic identity without any cell sorting. Treatment of mouse ESCs with these factors resulted in similar enhancements of myogenesis. These studies establish a foundation for serum-free and chemically defined monolayer skeletal myogenesis of ESCs.
Related JoVE Video
Dipeptidyl peptidase-4 inhibitors do not increase the risk of cardiovascular events in type 2 diabetes: a cohort study.
Acta Diabetol
PUBLISHED: 06-28-2014
Show Abstract
Hide Abstract
Two recent randomized controlled trials of type 2 diabetes mellitus (T2DM) patients with history of, or at high risk of, cardiovascular disease (CVD) showed no risk of ischemic cardiovascular events associated with dipeptidyl peptidase-4 inhibitors (DPP4i), but an increased risk of heart failure (HF) with saxagliptin. We evaluated the risk of CVD including myocardial infarction (MI), stroke, coronary revascularization, and HF associated with DPP4i in T2DM patients with and without baseline CVD as used in the community.
Related JoVE Video
Correlation Between Trainee Candidate Selection Criteria and Subsequent Performance.
J. Am. Coll. Surg.
PUBLISHED: 06-24-2014
Show Abstract
Hide Abstract
The objective of trainee recruitment is to identify candidates likely to perform well as trainees and subsequent faculty. The effectiveness of this process has not been established. The goal of this study was to identify trainee selection criteria predictive of excellent performance.
Related JoVE Video
Extraction, characterization and in vitro antioxidant activity of polysaccharides from black soybean.
Int. J. Biol. Macromol.
PUBLISHED: 06-23-2014
Show Abstract
Hide Abstract
Optimization of extraction conditions, preliminary characterization and in vitro antioxidant activity of polysaccharides from black soybean (BSPS) were investigated. The results of Box-Behnken design showed that the optimal extraction conditions for BSPS were as follows: ratio of water to material of 20ml/g, extraction time of 6.4h and extraction temperature of 92°C, with a corresponding yield of 2.56%. The crude BSPS were further fractionated on DEAE-52 and Sepharose CL-4B chromatography to afford three purified fractions (BSPS-1, BSPS-2 and BSPS-3). Chemical analysis showed that the three purified fractions were mainly composed of carbohydrate and uronic acid. In addition, BSPS-1 was composed of arabinose, rhamnose, galactose, glucose and mannose in the molar ratio of 1.79:1.00:2.59:26.54:1.01. BSPS-2 was composed of arabinose, rhamnose, xylose, galactose and mannose in the molar ratio of 8.10:4.80:9.15:13.38:1.00. BSPS-3 was composed of arabinose, rhamnose, galactose and mannose in the molar ratio of 16.80:3.60:33.66:1.00. The results of Fourier transform-infrared spectroscopy further confirmed the characteristic polysaccharide structures of the three purified fractions. Moreover, antioxidant assays showed crude BSPS and its purified fractions had potential superoxide anion and DPPH radical scavenging activities, and their antioxidant activity decreased in the order of crude BSPS>BSPS-3>BSPS-2>BSPS-1.
Related JoVE Video
Effects of Treg cells and IDO on human epithelial ovarian cancer cells under hypoxic conditions.
Mol Med Rep
PUBLISHED: 06-18-2014
Show Abstract
Hide Abstract
The aim of the present study was to explore the effect of hypoxia on ovarian cancer. A total of 6 samples were analyzed: SKOV3?IP cells (ovarian cancer cell line); SKOV3?IP and regulatory T (Treg) cells; SKOV3?IP and cytotoxic T lymphocytes (CTLs); SKOV3?IP and natural killer (NK) cells; SKOV3?IP co-cultured with CTLs and Treg cells; and SKOV3?IP co-cultured with Treg cells and NK cells. The expression of indoleamine 2,3?dioxygenase (IDO) was detected by reverse transcription-polymerase chain reaction (RT?PCR) and western blot analysis. An enzyme?linked immunosorbent assay (ELISA) was used to detect the concentration of transforming growth factor?? (TGF??), interferon?? (IFN??), interleukin?2 (IL?2), interleukin?10 (IL?10), and perforin. Moreover, ovarian cancer cell apoptosis and invasive ability were examined using flow cytometry and a Transwell chamber assay. IDO expression was significantly reduced in hypoxia and enhanced by Treg cells. Treg cells inhibited the IL?2, IFN?? and perforin secretion, and significantly (P<0.05) increased the IL?10 and TGF?? levels. The effects of Treg cells were enhanced with prolongation of the cell exposure to hypoxic conditions. In addition, Treg cells attenuated the promotive effect of CTLs and NK cells on cancer cell apoptosis. In addition, Treg cells significantly increased the SKOV3?IP invasive ability (P=0.00109) under hypoxic conditions. Our results suggest that IDO and Treg cells may serve as important therapeutic targets for patients with ovarian cancer.
Related JoVE Video
Long-term survival outcomes of video-assisted thoracic surgery for patients with non-small cell lung cancer.
Chin. J. Cancer Res.
PUBLISHED: 06-10-2014
Show Abstract
Hide Abstract
Video-assisted thoracic surgery (VATS) has been shown to be a safe alternative to conventional thoracotomy for patients with non-small cell lung cancer (NSCLC). However, popularization of this relatively novel technique has been slow, partly due to concerns about its long-term outcomes. The present study aimed to evaluate the long-term survival outcomes of patients with NSCLC after VATS, and to determine the significant prognostic factors on overall survival.
Related JoVE Video
Reduced oxidative stress contributes to the lipid lowering effects of isoquercitrin in free Fatty acids induced hepatocytes.
Oxid Med Cell Longev
PUBLISHED: 06-01-2014
Show Abstract
Hide Abstract
Oxidative stress interferes with hepatic lipid metabolism at various levels ranging from benign lipid storage to so-called second hit of inflammation activation. Isoquercitrin (IQ) is widely present flavonoid but its effects on hepatic lipid metabolism remain unknown. We used free fatty acids (FFA) induced lipid overload and oxidative stress model in two types of liver cells and measured cell viability, intracellular lipids, and reactive oxygen species (ROS) within hepatocytes. In addition, Intracellular triglycerides (TG), superoxide dismutase (SOD), and malondialdehyde (MDA) were examined. A novel in vitro model was used to evaluate correlation between lipid lowering and antioxidative activities. Furthermore, 34 major cytokines and corresponding ROS levels were analyzed in FFA/LPS induced coculture model between hepatocytes and Kupffer cells. At molecular level AMPK pathway was elucidated. We showed that IQ attenuated FFA induced lipid overload and ROS within hepatocytes. Further, IQ reversed FFA induced increase in intracellular TG SOD and MDA. It was shown that antioxidative activity of IQ correlates with its lipid lowering potentials. IQ reversed major proinflammatory cytokines and oxidative stress in FFA/LPS induced coculture model. Finally, AMPK pathway was found responsible for metabolic benefits at molecular level. IQ strikingly manifests antioxidative and related lipid lowering activities in hepatocytes.
Related JoVE Video
A double-center randomized head-to-head comparison of the frequency-altering AKE-1 capsule and Pillcam SB2.
J Interv Gastroenterol
PUBLISHED: 05-16-2014
Show Abstract
Hide Abstract
Capsule endoscopy is the first-line examination for small bowel disease. This study was aimed to compare the performance between the frequency-altering AKE-1 capsule (AKE) and the Pillcam SB2 (PSB) capsule.
Related JoVE Video
Long-term risk of late and very late stent thrombosis in patients treated with everolimus against paclitaxel-eluting stents: an updated meta-analysis.
Coron. Artery Dis.
PUBLISHED: 05-14-2014
Show Abstract
Hide Abstract
Everolimus-eluting stent (EES) reduces the risk of late and very late stent thrombosis (ST) in a number of randomized controlled trials (RCTs). However, the benefits have been variable.
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.