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Find video protocols related to scientific articles indexed in Pubmed.
The 5th Annual Shanghai Symposium on Clinical & Pharmaceutical Solutions through Analysis.
Bioanalysis
PUBLISHED: 11-11-2014
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The 5th Annual Shanghai Symposium on Clinical & Pharmaceutical Solutions through Analysis (CPSA Shanghai 2014) was held on 16-19 April 2014 in Shanghai, China. The meeting featured highly interactive events including diversified symposia, roundtable discussions, workshops, poster sessions and conference awards. There were over 220 participants from more than 10 countries, with 45 oral presentations and 42 posters presented. In addition, the meeting included one preconference workshop and a joint session with local bioanalytical and drug metabolism discussion groups.
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A retrospective and comparative study of inflammatory myofibroblastic tumor of the liver.
J. Gastroenterol. Hepatol.
PUBLISHED: 11-03-2014
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Inflammatory myofibroblastic tumor of the liver (IMTL) is a very rare benign disease with a good prognosis.
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One-step pyrolytic synthesis of nitrogen and sulfur dual-doped porous carbon with high catalytic activity and good accessibility to small biomolecules.
ACS Appl Mater Interfaces
PUBLISHED: 10-28-2014
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As one of promising catalysts that contain high density of active sites, N doped carbons have been extensively researched, while the reports for N, S dual-doped carbon materials are far less exhaustive. Herein, devoid of activation process and template, N, S dual-doped porous carbon (N-S-PC) was prepared for the first time via one-step pyrolysis of sodium citrate and cysteine. Possessing unique porous structure and large pore volume as well as good accessibility, N-S-PC demonstrates significantly improved electrocatalytic activity toward oxidation of ascorbic acid (AA), dopamine (DA), and uric acid (UA). In the coexisting system, the peak potential separation between AA and DA is up to 251 mV, which is much larger than for most of the other carbons. On the basis of large potential separation and high current response, selective and sensitive simultaneous determination of AA, DA, and UA was successfully accomplished by differential pulse voltammetry, displaying a linear response from 50 to 2000 ?M, from 0.1 to 50 ?M, and from 0.1 to 50 ?M with a detection limit (S/N = 3) of 0.78, 0.02, and 0.06 ?M. This work highlights the importance of N, S dual doping and hierarchical porous carbons for efficient catalysis.
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Overlapping and Distinct Molecular Determinants Dictating the Antiviral Activities of TRIM56 against Flaviviruses and Coronavirus.
J. Virol.
PUBLISHED: 09-24-2014
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The tripartite motif-containing (TRIM) proteins have emerged as a new class of host antiviral restriction factors, with several demonstrating roles in regulating innate antiviral responses. Of >70 known TRIMs, TRIM56 inhibits replication of bovine viral diarrhea virus, a ruminant pestivirus of the family Flaviviridae, but has no appreciable effect on vesicular stomatitis virus (VSV), a rhabdovirus. Yet the antiviral spectrum of TRIM56 remains undefined. In particular, how TRIM56 impacts human-pathogenic viruses is unknown. Also unclear are the molecular determinants governing the antiviral activities of TRIM56. Herein, we show that TRIM56 poses a barrier to infections by yellow fever virus (YFV), dengue virus serotype 2 (DENV2), and human coronavirus virus (HCoV) OC43 but not encephalomyocarditis virus (EMCV). Moreover, by engineering cell lines conditionally expressing various TRIM56 mutants, we demonstrated that TRIM56's antiflavivirus effects required both the E3 ligase activity that lies in the N-terminal RING domain and the integrity of its C-terminal portion, while the restriction of HCoV-OC43 relied upon the TRIM56 E3 ligase activity alone. Furthermore, TRIM56 was revealed to impair YFV and DENV2 propagation by suppressing intracellular viral RNA accumulation but to compromise HCoV-OC43 infection at a later step in the viral life cycle, suggesting that distinct TRIM56 domains accommodate differing antiviral mechanisms. Altogether, TRIM56 is a versatile antiviral host factor that confers resistance to YFV, DENV2, and HCoV-OC43 through overlapping and distinct molecular determinants.
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Phosphine-catalyzed [4+1] annulation of 1,3-(aza)dienes with maleimides: highly efficient construction of azaspiro[4.4]nonenes.
Chem. Commun. (Camb.)
PUBLISHED: 09-20-2014
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Phosphine-catalyzed [4+1] annulation of electron-deficient 1,3-dienes or 1,3-azadienes with maleimides has been successfully developed under very mild conditions, providing a convenient and highly efficient method for constructing 2-azaspiro[4.4]nonenes and 1,7-diazaspiro[4.4]nonenes. This reaction represents the first example of [4+1] cyclization between electron-deficient 4?-conjugated systems and non-allylic phosphorus ylides.
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Climate change impacts on crop yield: evidence from China.
Sci. Total Environ.
PUBLISHED: 08-30-2014
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When estimating climate change impact on crop yield, a typical assumption is constant elasticity of yield with respect to a climate variable even though the elasticity may be inconstant. After estimating both constant and inconstant elasticities with respect to temperature and precipitation based on provincial panel data in China 1980-2008, our results show that during that period, the temperature change contributes positively to total yield growth by 1.3% and 0.4% for wheat and rice, respectively, but negatively by 12% for maize. The impacts of precipitation change are marginal. We also compare our estimates with other studies and highlight the implications of the inconstant elasticities for crop yield, harvest and food security. We conclude that climate change impact on crop yield would not be an issue in China if positive impacts of other socio-economic factors continue in the future.
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Children's responses to mother-infant and father-infant interaction with a baby sibling: jealousy or joy?
J Fam Psychol
PUBLISHED: 08-25-2014
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Firstborn children's reactions to mother-infant and father-infant interaction after a sibling's birth were examined in an investigation of 224 families. Triadic observations of parent-infant-sibling interaction were conducted at 1 month after the birth. Parents reported on children's problem behaviors at 1 and 4 months after the birth and completed the Attachment Q-sort before the birth. Latent profile analysis (LPA) identified 4 latent classes (behavioral profiles) for mother-infant and father-infant interactions: regulated-exploration, disruptive-dysregulated, approach-avoidant, and anxious-clingy. A fifth class, attention-seeking, was found with fathers. The regulated-exploration class was the normative pattern (60%), with few children in the disruptive class (2.7%). Approach-avoidant children had more behavior problems at 4 months than any other class, with the exception of the disruptive children, who were higher on aggression and attention problems. Before the birth, anxious-clingy children had less secure attachments to their fathers than approach avoidant children but more secure attachments to their mothers. Results underscore individual differences in firstborns' behavioral responses to parent-infant interaction and the importance of a person-centered approach for understanding children's jealousy.
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Intake of dietary flavonoids and risk of epithelial ovarian cancer.
Am. J. Clin. Nutr.
PUBLISHED: 08-20-2014
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The impact of different dietary flavonoid subclasses on risk of epithelial ovarian cancer is unclear, with limited previous studies that have focused on only a few compounds.
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Dendritic Ionic Liquids Based on Imidazolium-Modified Poly(aryl ether) Dendrimers.
Chem Asian J
PUBLISHED: 08-16-2014
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A series of dendritic ionic liquids (DILs) based on imidazolium-modified poly(aryl ether) dendrimers IL-Br-Gn (n=0-3) were synthesized by a modified convergent approach and "click" chemistry. The resulting DILs exhibited high thermal resistance with decomposition temperatures up to 270?°C and low glass transition temperatures in the range of approximately -5-0?°C. All IL-Br-Gn were found to be miscible with water at any ratio and could encapsulate hydrophobic molecules. The reversible phase transfer of the DILs between the aqueous and organic phases was accomplished by simple anion exchange between the hydrophilic Br(-) anion and the hydrophobic bis(trifluoromethylsulfonyl)amide anion (NTf2 (-) ). IL-Br-Gn could be used as transporters to shuttle hydrophobic molecules between the organic and aqueous phases efficiently. The present work provides a new kind of transporting materials with potential applications in substance separation, drug delivery, and biomolecule transport.
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A Molecular Copper Catalyst for Electrochemical Water Reduction with a Large Hydrogen-Generation Rate Constant in Aqueous Solution.
Angew. Chem. Int. Ed. Engl.
PUBLISHED: 08-15-2014
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The copper complex [(bztpen)Cu](BF4 )2 (bztpen=N-benzyl-N,N',N'-tris(pyridin-2-ylmethyl)ethylenediamine) displays high catalytic activity for electrochemical proton reduction in acidic aqueous solutions, with a calculated hydrogen-generation rate constant (kobs ) of over 10000 s(-1) . A turnover frequency (TOF) of 7000 h(-1) ?cm(-2) and a Faradaic efficiency of 96?% were obtained from a controlled potential electrolysis (CPE) experiment with [(bztpen)Cu](2+) in pH 2.5 buffer solution at -0.90 V versus the standard hydrogen electrode (SHE) over two hours using a glassy carbon electrode. A mechanism involving two proton-coupled reduction steps was proposed for the dihydrogen generation reaction catalyzed by [(bztpen)Cu](2+) .
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Astrocytic metabolic and inflammatory changes as a function of age.
Aging Cell
PUBLISHED: 08-09-2014
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This study examines age-dependent metabolic-inflammatory axis in primary astrocytes isolated from brain cortices of 7-, 13-, and 18-month-old Sprague-Dawley male rats. Astrocytes showed an age-dependent increase in mitochondrial oxidative metabolism respiring on glucose and/or pyruvate substrates; this increase in mitochondrial oxidative metabolism was accompanied by increases in COX3/18SrDNA values, thus suggesting an enhanced mitochondrial biogenesis. Enhanced mitochondrial respiration in astrocytes limits the substrate supply from astrocytes to neurons; this may be viewed as an adaptive mechanism to altered cellular inflammatory-redox environment with age. These metabolic changes were associated with an age-dependent increase in hydrogen peroxide generation (largely ascribed to an enhanced expression of NOX2) and NF?B signaling in the cytosol as well as its translocation to the nucleus. Astrocytes also displayed augmented responses with age to inflammatory cytokines, IL-1?, and TNF?. Activation of NF?B signaling resulted in increased expression of nitric oxide synthase 2 (inducible nitric oxide synthase), leading to elevated nitric oxide production. IL-1? and TNF? treatment stimulated mitochondrial oxidative metabolism and mitochondrial biogenesis in astrocytes. It may be surmised that increased mitochondrial aerobic metabolism and inflammatory responses are interconnected and support the functionality switch of astrocytes, from neurotrophic to neurotoxic with age.
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A low level of GPR37 is associated with human hepatocellular carcinoma progression and poor patient survival.
Pathol. Res. Pract.
PUBLISHED: 08-09-2014
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GPR37, also known as parkin-associated endothelin-like receptor (Pael-R), is an orphan G protein-coupled receptor (GPCR). It has been reported that GPCRs play vital roles in the development and progression of cancer. To investigate the potential roles of GPR37 in hepatocellular carcinoma (HCC), expression of GPR37 was examined in human HCC samples. Immunohistochemistry and Western blot analyses were performed for GPR37 in 57 hepatocellular carcinoma samples. GPR37 expression was low in hepatocellular carcinoma as compared with the adjacent non-tumorous tissues. Clinicopathological analysis showed that GPR37 expression was significantly correlated with histological grade and the level of alpha fetal protein (AFP) (P=0.000 and 0.002, respectively). The Kaplan-Meier survival curves revealed that decreasing GPR37 expression was associated with poor prognosis in HCC patients, while in vitro, following the release from serum starvation of HuH7 HCC cell, the expression of GPR37 was downregulated. In addition, the transient GPR37 knockdown by siRNA in HuH7 cells significantly decreased the apoptosis of hepatoma cells with activation of the phosphatidylinositol 3-kinase-Akt signaling pathway. Our data suggest that GPR37 may play an important role in the pathogenesis of hepatocellular carcinoma by affecting the proliferation of H CC cells, and it could be a novel potential molecular therapy target for HCC.
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UBR5-mediated ubiquitination of ATMIN is required for ionizing radiation-induced ATM signaling and function.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 08-04-2014
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The Mre11/Rad50/NBS1 (MRN) protein complex and ATMIN protein mediate ATM kinase signaling in response to ionizing radiation (IR) and chromatin changes, respectively. NBS1 and ATMIN directly compete for ATM binding, but the molecular mechanism favoring either NBS1 or ATMIN in response to specific stimuli is enigmatic. Here, we identify the E3 ubiquitin ligase UBR5 as a key component of ATM activation in response to IR. UBR5 interacts with ATMIN and catalyzes ubiquitination of ATMIN at lysine 238 in an IR-stimulated manner, which decreases ATMIN interaction with ATM and promotes MRN-mediated signaling. We show that UBR5 deficiency, or mutation of ATMIN lysine 238, prevents ATMIN dissociation from ATM and inhibits ATM and NBS1 foci formation after IR, thereby impairing checkpoint activation and increasing radiosensitivity. Thus, UBR5-mediated ATMIN ubiquitination is a vital event for ATM pathway selection and activation in response to DNA damage.
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Blocking the function of inflammatory cytokines and mediators by using IL-10 and TGF-?: a potential biological immunotherapy for intervertebral disc degeneration in a beagle model.
Int J Mol Sci
PUBLISHED: 07-29-2014
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The debilitating effects of lower back pain are a major health issue worldwide. A variety of factors contribute to this, and oftentimes intervertebral disk degeneration (IDD) is an underlying cause of this disorder. Inflammation contributes to IDD, and inflammatory cytokines such as tumor necrosis factor (TNF)-? and interleukin (IL)-1?, play key roles in the pathology of IDD. Therefore, the development of treatments that inhibit the expression and/or effects of TNF-? and IL-1? in IDD patients should be a promising therapeutic approach to consider. This study characterized the potential to suppress inflammatory cytokine production in degenerative intervertebral disc (NP) cells by treatment with IL-10 and TGF-? in a canine model of IDD. IDD was induced surgically in six male beagles, and degenerative NP cells were isolated and cultured for in vitro studies on cytokine production. Cultured degenerative NP cells were divided into four experimental treatment groups: untreated control, IL-10-treated, TGF-?-treated, and IL-10- plus TGF-?-treated cells. Cultured normal NP cells served as a control group. TNF-? expression was evaluated by fluorescence activated cell sorting (FACS) analysis and enzyme-linked immunosorbent assay (ELISA); moreover, ELISA and real-time PCR were also performed to evaluate the effect of IL-10 and TGF-? on NP cell cytokine expression in vitro. Our results demonstrated that IL-10 and TGF-? treatment suppressed the expression of IL-1? and TNF-? and inhibited the development of inflammatory responses. These data suggest that IL-10 and TGF-? should be evaluated as therapeutic approaches for the treatment of lower back pain mediated by IDD.
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A family-oriented psychosocial intervention reduces inflammation in low-SES African American youth.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 07-21-2014
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Children of low socioeconomic status (SES) are at elevated risk for health problems across the lifespan. Observational studies suggest that nurturant parenting might offset some of these health risks, but their design precludes inferences about causal direction and clinical utility. Here we ask whether a psychosocial intervention, focused improving parenting, strengthening family relationships, and building youth competencies, can reduce inflammation in low-SES, African Americans from the rural South. The trial involved 272 mothers and their 11-y-old children from rural Georgia, half of whose annual household incomes were below the federal poverty line. Families were randomly assigned to a 7-wk psychosocial intervention or to a control condition. When youth reached age 19, peripheral blood was collected to quantify six cytokines that orchestrate inflammation, the dysregulation of which contributes to many of the health problems known to pattern by SES. Youth who participated in the intervention had significantly less inflammation on all six indicators relative to controls (all P values < 0.001; effect sizes in Cohen's d units ranged from -0.69 to -0.91). Mediation analyses suggested that improved parenting was partially responsible for the intervention's benefits. Inflammation was lowest among youth who received more nurturant-involved parenting, and less harsh-inconsistent parenting, as a consequence of the intervention. These findings have theoretical implications for research on resilience to adversity and the early origins of disease. If substantiated, they may also highlight a strategy for practitioners and policymakers to use in ameliorating social and racial health disparities.
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TLR9 is critical for glioma stem cell maintenance and targeting.
Cancer Res.
PUBLISHED: 07-21-2014
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Understanding supports for cancer stem-like cells in malignant glioma may suggest therapeutic strategies for their elimination. Here, we show that the Toll-like receptor TLR9 is elevated in glioma stem-like cells (GSC) in which it contributes to glioma growth. TLR9 overexpression is regulated by STAT3, which is required for GSC maintenance. Stimulation of TLR9 with a CpG ligand (CpG ODN) promoted GSC growth, whereas silencing TLR9 expression abrogated GSC development. CpG-ODN treatment induced Frizzled4-dependent activation of JAK2, thereby activating STAT3. Targeted delivery of siRNA into GSC was achieved via TLR9 using CpG-siRNA conjugates. Through local or systemic treatment, administration of CpG-Stat3 siRNA to silence STAT3 in vivo reduced GSC along with glioma growth. Our findings identify TLR9 as a functional marker for GSC and a target for the delivery of efficacious therapeutics for glioma treatment. Cancer Res; 74(18); 5218-28. ©2014 AACR.
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[Comparison of total disc replacement versus fusion for lumbar degenerative disc disease: a Meta-analysis of randomized controlled trials].
Zhonghua Wai Ke Za Zhi
PUBLISHED: 07-19-2014
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To compare the related clinical outcomes of total disc replacement (TDR) versus fusion in management of lumbar degenerative disc disease (LDDD)and provid available basis for choice of surgical procedure.
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Racial Microstressors, Racial Self-Concept, and Depressive Symptoms Among Male African Americans During the Transition to Adulthood.
J Youth Adolesc
PUBLISHED: 07-17-2014
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Racial discrimination is a pervasive stressor that can undermine mental health among African American youth and young adults. Several studies identify links between racial discrimination and depressive symptoms; however, this research base does not focus on male African American youth who experience significant racism-related stress during the transition to young adulthood. Moreover, few prospective studies consider significant confounding variables that affect exposure to and perception of discriminatory treatment. In response to this need, we examined the effect of exposure to racial discrimination from ages 16 to 18 on depressive symptoms among male African Americans at age 20. Racial self-concept, one's sense of positivity about one's race, was examined as a mediator and self-control as a moderator. Hypotheses were tested with 222 participants, age 16 at baseline and age 20 at the endpoint. Participants provided self-report data at five time points. Exposure to racial discrimination from ages 16 to 18 predicted depressive symptoms at age 20, net of confounding influences. Racial self-concept mediated this effect. Self-control moderated the influence of discrimination on racial self-concept. This study underscores the salience of racial discrimination in the development of depressive symptoms among African American male youth and the clinical utility of interventions targeting racial pride and self-control.
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Zero-velocity magnetophoretic method for the determination of particle magnetic susceptibility.
Anal Sci
PUBLISHED: 07-11-2014
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A simple zero-velocity method to determine the particle magnetic susceptibility by measuring the magnetophoretic velocity was proposed. The principle is that the magnetophoretic velocity of a particle in a liquid medium must be zero when the magnetic susceptibilities of the medium and the particle are equal, or the gravity force and the magnetophoretic force are balanced. By changing the medium magnetic susceptibility and measuring the magnetophoretic velocity of a particle, the particle magnetic susceptibility was determined from the medium magnetic susceptibility under the zero-velocity condition. The feasibility of the method was demonstrated for polystyrene particles using a Dy(III) solution in the horizontal migration mode and different organic solvents in the vertical migration mode.
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Global bioanalytical support.
Bioanalysis
PUBLISHED: 06-25-2014
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With the globalization of drug development, there is an increasing need for global bioanalytical support. Bioanalysis provides pivotal data for toxicokinetic, pharmacokinetic, bioavailability and bioequivalence studies used for regional or global regulatory submission. There are many known complications in building a truly global bioanalytical operation, ranging from lack of global regulatory guidelines and global standard operating procedures to barriers in regional requirements on sample shipping, importation and exportation. The primary objective of this article is to discuss common experiences and challenges facing the biopharmaceutical industry when providing bioanalytical support in a global setting. The key components of global bioanalytical services include the supporting infrastructure, spanning project management, IT support of data management, best practices in bioanalytical method transfer and sample analysis, and comprehensive knowledge of the requirements of bioanalysis guidelines and differences in these guidelines. A case study will highlight best practices for successful management of a global project.
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Reconstruction of lactate utilization system in Pseudomonas putida KT2440: a novel biocatalyst for l-2-hydroxy-carboxylate production.
Sci Rep
PUBLISHED: 06-24-2014
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As an important method for building blocks synthesis, whole cell biocatalysis is hindered by some shortcomings such as unpredictability of reactions, utilization of opportunistic pathogen, and side reactions. Due to its biological and extensively studied genetic background, Pseudomonas putida KT2440 is viewed as a promising host for construction of efficient biocatalysts. After analysis and reconstruction of the lactate utilization system in the P. putida strain, a novel biocatalyst that only exhibited NAD-independent d-lactate dehydrogenase activity was prepared and used in l-2-hydroxy-carboxylates production. Since the side reaction catalyzed by the NAD-independent l-lactate dehydrogenase was eliminated in whole cells of recombinant P. putida KT2440, two important l-2-hydroxy-carboxylates (l-lactate and l-2-hydroxybutyrate) were produced in high yield and high optical purity by kinetic resolution of racemic 2-hydroxy carboxylic acids. The results highlight the promise in biocatalysis by the biotechnologically important organism P. putida KT2440 through genomic analysis and recombination.
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Using Dynamic (99m)T c-GSA SPECT/CT Fusion Images for Hepatectomy Planning and Postoperative Liver Failure Prediction.
Ann. Surg. Oncol.
PUBLISHED: 06-24-2014
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Available tools in liver surgery planning rely on the future remnant liver (FRL) volume. Inappropriate decision might be made since the same FRL volume might represent different liver functions depending on the severity of underlying liver damage. This study developed an alternative system to estimate FRL function and to predict the risk of postoperative liver failure.
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Pathways of Peer Relationships from Childhood to Young Adulthood.
J Appl Dev Psychol
PUBLISHED: 06-21-2014
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This study examined trajectories of peer social preference during childhood and personality assessed in early adolescence in relation to trajectories of friendship quality during early adulthood. Participants (N = 585) were followed from age 5 to age 23. At ages 5 to 8, peers provided sociometric nominations; at age 12 participants reported their own personality characteristics; from age 19 to 23 participants rated their friendship quality. Latent growth modeling revealed that trajectories characterized by high levels of childhood peer social preference were related to trajectories characterized by high levels of early adulthood friendship quality. Early adolescent personality characterized by extraversion and conscientiousness predicted higher friendship quality at age 19, and conscientiousness predicted change in friendship quality from age 19 to 23. This study demonstrates that peer relationships show continuity from childhood to early adulthood and that qualities of core personality are linked to the development of adult friendships.
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Inhibition of glycogen synthase kinase-3? attenuates acute kidney injury in sodium taurocholate?induced severe acute pancreatitis in rats.
Mol Med Rep
PUBLISHED: 06-11-2014
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The aim of the present study was to investigate the efficacy of 4?benzyl?2?methyl?1,2,4?thiadiazolidine?3,5?dione (TDZD?8), the selective inhibitor of glycogen synthase kinase?3? (GSK?3?), on the development of acute kidney injury in an experimental model of sodium taurocholate?induced severe acute pancreatitis (SAP) in rats. The serum amylase, lipase, interleukin?1? and interleukin?6 levels, and the pancreatic pathological score were examined to determine the magnitude of pancreatitis injury. The serum creatinine and blood urea nitrogen levels, myeloperoxidase (MPO) activity and renal histological grading were measured to assess the magnitude of SAP?induced acute kidney injury. The activation of nuclear factor??B (NF??B) was examined using an immunohistochemistry assay. The expression of GSK?3?, phospho?GSK?3? (Ser9), tumour necrosis factor?? (TNF??), intercellular adhesion molecule?1 (ICAM?1) and inducible nitric oxide synthase (iNOS) protein in the kidney was characterised using western blot analysis. TDZD?8 attenuated (i) serum amylase, lipase and renal dysfunction; (ii) the serum concentrations of proinflammatory cytokines; (iii) pancreatic and renal pathological injury; (iv) renal MPO activity and (v) NF??B activation and TNF??, ICAM?1 and iNOS protein expression in the kidney. The results obtained in the present study suggest that the inhibition of GSK?3? attenuates renal disorders associated with SAP through the inhibition of NF??B activation and the downregulation of the expression of proinflammatory cytokines, TNF??, ICAM?1 and iNOS in rats. Blocking GSK?3? protein kinase activity may be a novel approach to the treatment of this in?ammatory condition.
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The mechanism for increasing the oral bioavailability of poorly water-soluble drugs using uniform mesoporous carbon spheres as a carrier.
Drug Deliv
PUBLISHED: 05-30-2014
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Abstract Uniform mesoporous carbon spheres (UMCS) were used as a carrier to improve the bioavailability of the model drug, celecoxib (CEL). Furthermore, we investigated the mechanism responsible for the improved bioavailability of CEL. The association, adhesion and uptake of UMCS by intestinal epithelial cells were studied by transmission electron microscopy (TEM), fluorescence-activated cell sorting (FACS) and laser confocal scanning microscopy (LCSM). UMCS was found to promote cellular uptake of CEL. Drug transport in Caco-2 cell monolayers proved that UMCS can significantly reduce the rate of drug efflux and improve CEL permeability. The dissolution rate of CEL from drug-loaded samples was markedly improved compared with pure crystalline CEL; moreover, oral bioavailability of CEL loaded into UMCS was also markedly improved compared with that of commercially available capsules. UMCS indicates the advantages and potential of this method to achieve improved oral absorption by increasing the dissolution rate, cellular uptake and permeability of the drug.
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Hyperactivated Wnt signaling induces synthetic lethal interaction with Rb inactivation by elevating TORC1 activities.
PLoS Genet.
PUBLISHED: 05-01-2014
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Inactivation of the Rb tumor suppressor can lead to increased cell proliferation or cell death depending on specific cellular context. Therefore, identification of the interacting pathways that modulate the effect of Rb loss will provide novel insights into the roles of Rb in cancer development and promote new therapeutic strategies. Here, we identify a novel synthetic lethal interaction between Rb inactivation and deregulated Wg/Wnt signaling through unbiased genetic screens. We show that a weak allele of axin, which deregulates Wg signaling and increases cell proliferation without obvious effects on cell fate specification, significantly alters metabolic gene expression, causes hypersensitivity to metabolic stress induced by fasting, and induces synergistic apoptosis with mutation of fly Rb ortholog, rbf. Furthermore, hyperactivation of Wg signaling by other components of the Wg pathway also induces synergistic apoptosis with rbf. We show that hyperactivated Wg signaling significantly increases TORC1 activity and induces excessive energy stress with rbf mutation. Inhibition of TORC1 activity significantly suppressed synergistic cell death induced by hyperactivated Wg signaling and rbf inactivation, which is correlated with decreased energy stress and decreased induction of apoptotic regulator expression. Finally the synthetic lethality between Rb and deregulated Wnt signaling is conserved in mammalian cells and that inactivation of Rb and APC induces synergistic cell death through a similar mechanism. These results suggest that elevated TORC1 activity and metabolic stress underpin the evolutionarily conserved synthetic lethal interaction between hyperactivated Wnt signaling and inactivated Rb tumor suppressor.
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Counting of E. Coli by a micro-flow cytometer based on a photonic-microfluidic integrated device.
Electrophoresis
PUBLISHED: 04-21-2014
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Counting of E. coli DH5? cell suspensions in phosphate buffered saline is performed using a micro-flow cytometer based on a photonic-microfluidic integrated device. Side-scattered light signals are used to count the E. coli cells. A detection efficiency of 92% is achieved when compared with the expected count from a haemocytometer. The detection efficiency is correlated to the ratio of sample to sheath flow rates. It is demonstrated that E. coli can be easily distinguished from beads of similar sizes (2-4?m) as their scattering intensities are different. This article is protected by copyright. All rights reserved.
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Etanercept plus topical corticosteroids as initial therapy for grade one acute graft-versus-host disease after allogeneic hematopoietic cell transplantation.
Biol. Blood Marrow Transplant.
PUBLISHED: 04-18-2014
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Clinical diagnosis of grade 1 acute graft-versus-host disease (GVHD) marks the beginning of a potentially progressive and fatal course of GVHD after hematopoietic stem cell transplantation (HSCT). However, interventional studies to treat early GVHD are lacking. We conducted a single-arm prospective phase II trial to test the hypothesis that treatment of newly diagnosed grade 1 acute GVHD with etanercept and topical corticosteroids would reduce progression to grade 2 to 4 within 28 days. Study patients (n = 34) had a median age of 51 years (range, 10 to 67 years) and had undergone unrelated (n = 22) or related (n = 12) donor HSCT. Study patients were treated with etanercept (.4 mg/kg, maximum 25 mg/dose) twice weekly for 4 to 8 weeks. Ten of 34 patients (29%) progressed to grade 2 to 4 acute GVHD within 28 days. The cumulative incidence of grade 2 to 4 and grade 3 to 4 acute GVHD at 1 year was 41% and 3%, respectively. Nonrelapse mortality was 19% and overall survival was 63% at 2 years. Among a contemporaneous control cohort of patients who were diagnosed with grade 1 acute GVHD and treated with topical corticosteroids but not etanercept during the study period, 12 of 28 patients (43%) progressed to grade 2 to 4 GVHD within 28 days, with a 1-year incidence of grade 2 to 4 GVHD and grade 3 to 4 GVHD of 61% (41% versus 61%, P = .08) and 18% (3% versus 18%, P = .05), respectively. Patients treated with etanercept also experienced less increase in GVHD plasma biomarkers suppression of tumorigenicity 2 (P = .06) and regenerating islet-derived 3-alpha (P = .01) 28 days after grade 1 acute GVHD diagnosis compared with contemporaneous control patients. This study was terminated early because of poor accrual. Future prospective studies are needed to identify patients with grade 1 acute GVHD at risk of swift progression to more severe GVHD and to establish consensus for the treatment of grade 1 acute GVHD. This trial is registered with ClinicalTrials.gov, number NCT00726375.
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Long non-coding RNA 91H contributes to the occurrence and progression of esophageal squamous cell carcinoma by inhibiting IGF2 expression.
Mol. Carcinog.
PUBLISHED: 04-08-2014
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Long non-coding RNAs (lncRNAs) have been recently recognized as a major class of regulators in mammalian systems. 91H, a novel long noncoding antisense transcripts located on the position of the H19/IGF2 locus has been suggested to play a potential tumor-suppressor role in tumor development. However, little study has proved the mechanism in esophageal squamous cell carcinoma (ESCC). We carried out this study to explore the role of lncRNA 91H in the regulation of H19 imprinting control regions (ICR) and IGF2 expression and the association between 91H and ESCC progression. The cell line TE-1, Eca-109, and 232 ESCC patients' matched sets of paraffin-embedded adjacent normal and tumor samples were obtained in this study. The results showed that 91H expression was significantly lower in patients with higher depth of invasion, neoplastic grading and TNM which usually leads to the overexpression of IGF2 in tumor progression. The expression of 91H usually decreased in TE-1 and Eca-109 when treated with demethylation agent. Further analysis revealed that, in 91H knockdown cell lines, IGF2 expression was also significantly higher than negative controls. Therefore, the results demonstrated that the lncRNA 91H was associated with H19 ICR methylation and inhibited IGF2 expression of ESCC patients which may optimize the mechanism of IGF2 regulation in tumor development. Patients with higher depth of invasion, neoplastic grading and TNM usually demonstrated lower 91H expression potentially represent a novel clinically relevant event to identify individuals at increased risk for the occurrence, progression and prognosis of ESCC. © 2014 Wiley Periodicals, Inc.
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Chondrogenic differentiation of adipose-derived stromal cells in combinatorial hydrogels containing cartilage matrix proteins with decoupled mechanical stiffness.
Tissue Eng Part A
PUBLISHED: 04-07-2014
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Adipose-derived stromal cells (ADSCs) are attractive autologous cell sources for cartilage repair given their relative abundance and ease of isolation. Previous studies have demonstrated the potential of extracellular matrix (ECM) molecules as three-dimensional (3D) scaffolds for promoting chondrogenesis. However, few studies have compared the effects of varying types or doses of ECM molecules on chondrogenesis of ADSCs in 3D. Furthermore, increasing ECM molecule concentrations often result in simultaneous changes in the matrix stiffness, which makes it difficult to elucidate the relative contribution of biochemical cues or matrix stiffness on stem cell fate. Here we report the development of an ECM-containing hydrogel platform with largely decoupled biochemical and mechanical cues by modulating the degree of methacrylation of ECM molecules. Specifically, we incorporated three types of ECM molecules that are commonly found in the cartilage matrix, including chondroitin sulfate (CS), hyaluronic acid (HA), and heparan sulfate (HS). To elucidate the effects of interactive biochemical and mechanical signaling on chondrogenesis, ADSCs were encapsulated in 39 combinatorial hydrogel compositions with independently tunable ECM types (CS, HA, and HS), concentrations (0.5%, 1.25%, 2.5%, and 5% [w/v]), and matrix stiffness (3, 30, and 90?kPa). Our results show that the effect of ECM composition on chondrogenesis is dependent on the matrix stiffness of hydrogels, suggesting that matrix stiffness and biochemical cues interact in a nonlinear manner to regulate chondrogenesis of ADSCs in 3D. In soft hydrogels (~3 kPa), increasing HA concentrations resulted in substantial upregulation of aggrecan and collagen type II expression in a dose-dependent manner. This trend was reversed in HA-containing hydrogels with higher stiffness (~90?kPa). The platform reported herein could provide a useful tool for elucidating how ECM biochemical cues and matrix stiffness interact together to regulate stem cell fate, and for rapidly optimizing ECM-containing scaffolds to support stem cell differentiation and tissue regeneration.
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Microbial lactate utilization: enzymes, pathogenesis, and regulation.
Trends Microbiol.
PUBLISHED: 03-23-2014
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Lactate utilization endows microbes with the ability to use lactate as a carbon source. Lactate oxidizing enzymes play key roles in the lactate utilization pathway. Various types of these enzymes have been characterized, but novel ones remain to be identified. Lactate determination techniques and biocatalysts have been developed based on these enzymes. Lactate utilization has also been found to induce pathogenicity of several microbes, and the mechanisms have been investigated. More recently, studies on the structure and organization of operons of lactate utilization have been carried out. This review focuses on the recent progress and future perspectives in understanding microbial lactate utilization.
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The diplotype Fas -1377A/-670G as a genetic marker to predict a lower risk of breast cancer in Chinese women.
Tumour Biol.
PUBLISHED: 03-11-2014
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This study was designed to reveal the effects of Fas and FasL polymorphisms of interest on breast cancer risk. A total of 439 patients with breast cancer and 439 controls were enrolled in this study. The genotypes Fas -1377G/A, Fas -670A/G, and FasL -844 T/C were detected by MassARRAY. The protein expressions of estrogen receptor, progesterone receptor, and CerbB-2 were determined by immunohistochemistry. Among the 439 patients, Fas mRNA levels in 22 samples of breast cancer and adjacent normal tissues were detected by real-time polymerase chain reaction, and the soluble Fas and Fas ligand concentrations of 180 patients were measured by enzyme-linked immunosorbent assay. The Fas -1377GA, Fas -1377AA, Fas -670AG, Fas -670GG, and FasL -844TC genotypes were associated with a reduced risk of breast cancer. Haplotype analysis indicated that Fas -1377G/-670A was associated with an increased risk of breast cancer, whereas Fas -1377A/-670A was associated with the opposite effect. Furthermore, gene-gene interaction analysis revealed that the Fas -1377GA/AA (-670AG/GG) and FasL -844CC or TC/TT genotypes were associated with a decreased risk of breast cancer. Meanwhile, -1377GG and -670AA genotypes were associated with higher soluble Fas concentrations than other genotypes. We conclude that Fas and FasL polymorphisms can affect breast cancer risk and that Fas polymorphisms are likely to affect breast cancer risk by regulating the soluble Fas concentration.
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Intraclass diversification of immunoglobulin heavy chain genes in the African lungfish.
Immunogenetics
PUBLISHED: 03-10-2014
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Lungfish (Dipnoi) are the closest living relatives to tetrapods, and they represent the transition from water to land during vertebrate evolution. Lungfish are armed with immunoglobulins (Igs), one of the hallmarks of the adaptive immune system of jawed vertebrates, but only three Ig forms have been characterized in Dipnoi to date. We report here a new diversity of Ig molecules in two African lungfish species (Protopterus dolloi and Protopterus annectens). The African lungfish Igs consist of three IgMs, two IgWs, three IgNs, and an IgQ, where both IgN and IgQ originated evidently from the IgW lineage. Our data also suggest that the IgH genes in the lungfish are organized in a transiting form from clusters (IgH loci in cartilaginous fish) to a translocon configuration (IgH locus in tetrapods). We propose that the intraclass diversification of the two primordial gnathostome Ig classes (IgM and IgW) as well as acquisition of new isotypes (IgN and IgQ) has allowed lungfish to acquire a complex and functionally diverse Ig repertoire to fight a variety of microorganisms. Furthermore, our results support the idea that "tetrapod-specific" Ig classes did not evolve until the vertebrate adaptation to land was completed ~360 million years ago.
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Prevention Effects Ameliorate the Prospective Association Between Nonsupportive Parenting and Diminished Telomere Length.
Prev Sci
PUBLISHED: 03-07-2014
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Telomere length (TL) is an indicator of general systemic aging, with diminished TL associated with several chronic diseases of aging and with heightened mortality risk. Research has begun to focus on the ways in which stress contributes to telomere attrition. The purposes of this study were (a) to establish whether exposure to nonsupportive parenting, defined as high levels of conflict and rancor with low levels of warmth and emotional support, at age 17 would forecast TL 5 years later; and (b) to determine whether participation in an efficacious family-centered prevention program could ameliorate any associations that emerged. Rural African American adolescents participated in the Adults in the Making (AIM) program or a control condition. Primary caregivers provided data on nonsupportive parenting during a pretest when adolescents were age 17. Adolescents provided data on anger at the pretest and at a posttest administered 7 months later. When the youths were age 22, TL was assayed from a blood draw. The results indicated that heightened nonsupportive parenting forecast diminished TL among young adults in the control condition but not among those who participated in AIM; socioeconomic status risk, life stress, and the use of alcohol and cigarettes at age 17, and blood pressure and body mass index at age 22, were controlled. Subsequent exploratory analyses suggested that AIM-induced reductions in adolescents' anger served as a mediator connecting group assignment to TL. The results suggest that the cellular-level sequelae of nonsupportive parenting and stress are not immutable.
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Prevention moderates associations between family risks and youth catecholamine levels.
Health Psychol
PUBLISHED: 03-03-2014
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The purpose of this study was to establish, using a quasi-experimental design, whether 2 family risk factors, parental psychological dysfunction and nonsupportive parenting, during preadolescence could longitudinally predict elevated sympathetic nervous system (SNS) activity 9 years later, and to determine whether participation in an efficacious family centered prevention program could moderate these associations if they emerged.
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RVG-peptide-linked trimethylated chitosan for delivery of siRNA to the brain.
Biomacromolecules
PUBLISHED: 02-27-2014
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In this work, a peptide derived from the rabies virus glycoprotein (RVG) was linked to siRNA/trimethylated chitosan (TMC) complexes through bifunctional PEG for efficient brain-targeted delivery of siRNA. The physiochemical properties of the complexes, such as siRNA complexing ability, size and ? potential, morphology, serum stability, and cytotoxicity, were investigated prior to studying the cellular uptake, in vitro gene silencing efficiency, and in vivo biodistribution. The RVG-peptide-linked siRNA/TMC-PEG complexes showed increased serum stability, negligible cytotoxicity, and higher cellular uptake than the unmodified siRNA/TMC-mPEG complexes in acetylcholine receptor positive Neuro2a cells. The potent knockdown of BACE1, a therapeutic target in Alzheimer's disease, demonstrated the gene silencing efficiency. In vivo imaging analysis showed significant accumulation of Cy5-siRNA in the isolated brain of mice injected with RVG-peptide-linked complexes. Therefore, the RVG-peptide-linked TMC-PEG developed in this study can be used as a potential carrier for delivery of siRNA to the brain.
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Astrocyte transplantation for spinal cord injury: current status and perspective.
Brain Res. Bull.
PUBLISHED: 02-22-2014
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Spinal cord injury (SCI) often causes incurable neurological dysfunction because axonal regeneration in adult spinal cord is rare. Astrocytes are gradually recognized as being necessary for the regeneration after SCI as they promote axonal growth under both physiological and pathophysiological conditions. Heterogeneous populations of astrocytes have been explored for structural and functional restoration. The results range from the early variable and modest effects of immature astrocyte transplantation to the later significant, but controversial, outcomes of glial-restricted precursor (GRP)-derived astrocyte (GDA) transplantation. However, the traditional neuron-centric view and the concerns about the inhibitory roles of astrocytes after SCI, along with the sporadic studies and the lack of a comprehensive review, have led to some confusion over the usefulness of astrocytes in SCI. It is the purpose of the review to discuss the current status of astrocyte transplantation for SCI based on a dialectical view of the context-dependent manner of astrocyte behavior and the time-associated characteristics of glial scarring. Critical issues are then analyzed to reveal the potential direction of future research.
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Perceived discrimination among African American adolescents and allostatic load: a longitudinal analysis with buffering effects.
Child Dev
PUBLISHED: 02-05-2014
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This study was designed to examine the prospective relations of perceived racial discrimination with allostatic load (AL), along with a possible buffer of the association. A sample of 331 African Americans in the rural South provided assessments of perceived discrimination from ages 16 to 18 years. When youth were 18 years, caregivers reported parental emotional support and youth assessed peer emotional support. AL and potential confounder variables were assessed when youth were 20. Latent growth mixture modeling identified two perceived discrimination classes: high and stable, and low and increasing. Adolescents in the high and stable class evinced heightened AL even with confounder variables controlled. The racial discrimination to AL link was not significant for young adults who received high emotional support.
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Gender and racial/ethnic differences in the associations of urinary phthalate metabolites with markers of diabetes risk: National Health and Nutrition Examination Survey 2001-2008.
Environ Health
PUBLISHED: 02-05-2014
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Phthalates are ubiquitous endocrine disrupting chemicals associated with diabetes. Although women and minorities are more likely to be exposed to phthalates, no prior studies have examined phthalate exposure and markers of diabetes risk evaluating effect modification by gender and race/ethnicity.
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Catecholamine levels and delay discounting forecast drug use among African American youths.
Addiction
PUBLISHED: 02-03-2014
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To test hypotheses about the contributions of the catecholamines epinephrine and norepinephrine [which serve as biological markers of life stress through sympathetic nervous system (SNS) activation], delay discounting and their interaction to the prediction of drug use among young African American adults.
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Enantioselective organocatalytic oxaziridination of N-tosyl aldimine catalyzed by Cinchona alkaloid-ester derivatives.
Chirality
PUBLISHED: 01-22-2014
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A series of cinchona alkaloid-ester derivatives was synthesized and applied to catalyze the enantioselective oxaziridination of aryl aldimines with m-CPBA. The (R,R)-oxaziridines were obtained in good yields with high enantiomeric excess (ee) values (up to 98%).
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Theoretical studies on a new high energy density compound 6-amino-7-nitropyrazino[2,3-e][1,2,3,4]tetrazine 1,3,5-trioxide (ANPTTO).
J Mol Model
PUBLISHED: 01-17-2014
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The derivatives of 1,2,3,4-tetrazine may be promising candidates for high-energy density compounds and are receiving more and more attentions. In this study, a new derivative 6-amino-7-nitropyrazino[2,3-e][1,2,3,4]tetrazine 1,3,5-trioxide (ANPTTO) has been designed. The geometrical structure and IR spectrum in the gas phase were studied at the B3LYP/6-31G* level of density functional theory (DFT). The crystal structure was predicted by molecular mechanics method and refined by the GGA/BOP function of periodic DFT with the basis set of TNP. The gas phase enthalpy of formation was calculated by the homodesmotic reaction method. The enthalpy of sublimation and solid phase enthalpy of formation were also predicted. The detonation properties were estimated with the Kamlet-Jacobs equations based on the predicted density and enthalpy of formation in solid state. The available free space in the lattice and resonance energy were calculated to evaluate its stability. ANPTTO has a high stability and is a promising high energetic component with the density?>2 g?·?cm(-3), detonation velocity?>9000 m?·?s(-1), and detonation pressure?>40 GPa. A synthetic route was proposed to provide a consideration for further study.
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Computational investigations into the substituent effects of -N?, -NF?, -NO?, and -NH? on the structure, sensitivity and detonation properties of N, N'-azobis(1,2,4-triazole).
J Mol Model
PUBLISHED: 01-15-2014
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A series of derivatives of N, N'-azobis(1,2,4-triazole) substituted by -N?, -NF?, -NO?, and -NH? groups was studied using the density functional theory method. To reveal the orbital interactions clearly and interpret the stability of the title compounds, natural bonding orbital (NBO) analysis was carried out. Strong p-? and ?-? conjugation interactions exist in molecules. Substituent effects on the geometrical and electronic structures, aromaticity of the triazole ring, electronic sensitivity, impact sensitivity, thermal stability, density, solid state heat of formation [?H(f)(s)], detonation velocity (D), detonation pressure (P), and specific impulse (I(s)) were investigated. Substituent groups have significant and differing effects on performance. -N?, -NF?, and -NO? groups are very helpful for enhancing D and P, but the case is different for the -NH? group. The order of the contribution of various groups to P and D is -NF?>?-NO??>?-N??>?-NH?. -NF? brings the highest D and P, but the lowest I(s). -NO? results in the secondary highest D and P and the best electronic stability.-N? gives relatively low D, P and stability, but the highest ?H(f)(s) and I(s). -NH? leads to the lowest D and P, while giving the best impact and thermal stabilities. Therefore, it is necessary to consider various aspects comprehensively according to the practical requirements for each compound designed. Taking both detonation performance and sensitivity into consideration, introducing -NH? and -N? into N, N'-azobis(1, 2, 4-triazole) may be a good choice for designing high-energy density materials.
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Specific growth inhibition of ErbB2?expressing human breast cancer cells by genetically modified NK?92 cells.
Oncol. Rep.
PUBLISHED: 01-13-2014
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The natural killer cell line NK?92 shows great cytotoxicity against various types of cancer. Several types of solid tumor cells, however, can effectively resist NK-mediated lysis by interaction of major histocompatibility complex (MHC) molecules with NK cell inhibitory receptors. To generate a eukaryotic expression vector encoding chimeric antigen receptor scFv anti-erbB2-CD28-? and to investigate the expression and action of this chimeric antigen receptor in cancer cells both in vitro and in vivo, NK?92 cells were genetically modified with an scFv anti-erbB2-CD28-? chimeric recep-tor by optimized electro-poration using the Amaxa Nucleofector system. The expression of the chimeric receptor was evaluated by RT-PCR and immunofluorescence. The ability of the genetically modified NK?92 cells to induce cell death in tumor targets was assessed in vitro and in vivo. The transduced NK?92-anti-erbB2 scFv-CD28-? cells expressing high levels of the fusion protein on the cell surface were analyzed by fluorescence-activated cell-sorting (FACS) analysis. These cells specifically enhanced the cell death of the erbB2?expressing human breast cancer cell lines MDA-MB-453 and SKBr3. Furthermore, adoptive transfer of genetically modified NK?92 cells specifically reduced tumor size and lung metastasis of nude mice bearing established MDA-MB-453 cells, and significantly enhanced the survival period of these mice. The genetically modified NK?92 cells significantly enhanced the killing of erbB2?expressing cancer and may be a novel therapeutic strategy for erbB2?expressing cancer cells.
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Systematic review and meta-analysis on vitamin D receptor polymorphisms and cancer risk.
Tumour Biol.
PUBLISHED: 01-10-2014
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The vitamin D receptor (VDR) can influence cancer susceptibility through binding to vitamin D. However, the previous studies were contradictory. Therefore this meta-analysis was conducted to clarify the association between VDR polymorphisms (BsmI, TaqI, FokI, and ApaI) and cancer risk. One hundred twenty-six studies were enrolled through PubMed. For VDR BsmI polymorphism, significantly increased cancer risks were observed in the overall analysis. In the further stratified analysis, increased risks were observed in colorectal and skin cancer, especially in Caucasian population. However, no significant associations were observed in other VDR polymorphisms in the overall analysis. In the further subgroup analysis, increased risks were found in oral, breast, and basal cell cancer while decreased risk was found in prostate cancer in t allele carriers of TaqI polymorphism. For VDR FokI polymorphism, increased risks were found in ovarian and skin cancer while decreased risk in glioma in f allele carriers. For VDR ApaI polymorphism, increased risk was observed in basal cell cancer, especially in Asian population in a allele carriers. In conclusion, these results indicated that b allele of BamI polymorphism was a risk factor for cancer susceptibility. Meanwhile, t allele of TaqI polymorphism was a risk factor for oral, breast, and basal cell cancer and a protective factor for prostate cancer. Moreover, f allele of FokI polymorphism was a risk factor for ovarian and skin cancer and a protective factor for glioma. Finally, a allele of ApaI polymorphism was a risk factor for basal cell cancer in Asian population.
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Chaperonin containing TCP1, subunit 8 (CCT8) is upregulated in hepatocellular carcinoma and promotes HCC proliferation.
APMIS
PUBLISHED: 01-02-2014
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The development of molecular pathogenesis of hepatocellular carcinoma (HCC) is complex and involves alterations in the expression and conformation of assorted oncoproteins and tumor suppressors. Chaperonin containing TCP1 (CCT) is a cytolic molecular chaperone complex that is required for the correct folding of numerous proteins. In this study, we investigated a possible involvement of CCT subunit 8 (CCT8) in HCC development. Immunohistochemical analysis was performed in 102 human HCC samples. High CCT8 expression was detected in clinical HCC samples compared with adjacent noncancerous tissues. The univariate and multivariate survival analyses were also performed to determine their prognostic significance. Western blot confirmed the high expression of CCT8 in HCC compared with adjacent normal tissue. Moreover, the biological significance of the aberrant expression of CCT8 was investigated in HCC cell lines. Expression of CCT8 was correlated directly with the histologic grades and tumor size of HCC and high expression of CCT8 was associated with a poor prognosis. CCT8 depletion by siRNA inhibited cell proliferation and blocked S-phase entry in HuH7 cells. These results suggested that CCT8 might be an oncogene and participate in HCC cell proliferation. These findings provide a potential therapeutic strategy for the treatment of HCC.
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Three dimensional simulation of transport and fate of oil spill under wave induced circulation.
Mar. Pollut. Bull.
PUBLISHED: 01-02-2014
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An oil spill model is developed and coupled to a current-wave model to simulate oil spill transport in aquatic environments where waves are present. The oil spill model incorporates physical-chemical processes of oil spill, and simulates oil slick transport by a circulation-driven Lagrangian Parcel model. Using the coupled oil spill model and the current-wave model CH3D-SWAN, a laboratory observed wave induced circulation and oil slick evolution are successfully simulated, while different current-wave coupling schemes generate different flow patterns and oil slick evolution. The modeling system is also shown to simulate Langmuir circulation and resulting oil slicks. Hypothetical scenarios of oil spill near Virginia coast during Hurricane Isabel and Irene are simulated using the oil spill model and the CH3D-Storm Surge Modeling System to assess the role of storm waves during oil spill. The spill area is significantly larger when storm waves are considered, implying waves significantly increase oil spill dispersion.
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Enhanced anticancer activity of DM1-loaded star-shaped folate-core PLA-TPGS nanoparticles.
Nanoscale Res Lett
PUBLISHED: 01-01-2014
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The efficient delivery of therapeutic drugs into interested cells is a critical challenge to broad application of nonviral vector systems. In this research, emtansine (DM1)-loaded star-shaped folate-core polylactide-d-?-tocopheryl polyethylene glycol 1000 succinate (FA-PLA-TPGS-DM1) copolymer which demonstrated superior anticancer activity in vitro/vivo in comparison with linear FA-PLA-TPGS nanoparticles was applied to be a vector of DM1 for FR(+) breast cancer therapy. The DM1- or coumarin 6-loaded nanoparticles were fabricated, and then characterized in terms of size, morphology, drug encapsulation efficiency, and in vitro drug release. And the viability of MCF-7/HER2 cells treated with FA-DM1-nanoparticles (NPs) was assessed. Severe combined immunodeficient mice carrying MCF-7/HER2 tumor xenografts were treated in several groups including phosphate-buffered saline control, DM1, DM1-NPs, and FA-DM1-NPs. The antitumor activity was then assessed by survival time and solid tumor volume. All the specimens were prepared for formalin-fixed and paraffin-embedded tissue sections for hematoxylin-eosin staining. The data showed that the FA-DM1-NPs could efficiently deliver DM1 into MCF-7/HER2 cells. The cytotoxicity of DM1 to MCF-7/HER2 cells was significantly increased by FA-DM1-NPs when compared with the control groups. In conclusion, the FA-DM1-NPs offered a considerable potential formulation for FR(+) tumor-targeting biotherapy.
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Prognostic Value of Long Non-Coding RNA HOTAIR in Various Cancers.
PLoS ONE
PUBLISHED: 01-01-2014
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Long non-coding RNA has been involved in cancer progression, and high HOX transcript antisense intergenic RNA (HOTAIR) is thought to be a poor prognostic indicator in tumorigenesis of multiple types of cancer. Hence, the present study further reveals its prognostic value in tumor malignancy. A systematic review of PubMed and Web of Science was carried out to select literatures relevant to the correlation between HOTAIR expression levels and clinical outcome of various tumors. Overall survival (OS), metastasis-free survival (MFS), recurrence-free survival (RFS), and disease-free survival (DFS) were subsequently analyzed. Data from studies directly reporting a hazard ratio (HR) and the corresponding 95% confidence interval (CI) or a P value as well as survival curves were pooled in the current meta-analysis. A total of 2255 patients from 19 literatures almost published in 2011 or later were included in the analysis. The results suggest that HOTAIR was highly associated with HR for OS of 2.33 (95%CI?=?1.77-3.09, Pheterogeneity?=?0.016). Stratified analyses indicate that elevated levels of HOTAIR appears to be a powerful prognostic biomarker for patients with colorectal cancer (HR?=?3.02, 95CI%?=?1.84-4.95, Pheterogeneity?=?0.699) and esophageal squamous cell carcinomas (HR?=?2.24, 95CI%?=?1.67-3.01, Pheterogeneity?=?0.711), a similar effect was also observed in analysis method and specimen, except for ethnicity. In addition, Hazard ratios for up-regulation of HOTAIR for MFS, RFS, and DFS were 2.32 (P<0.001), 1.98 (P?=?0.369), and 3.29 (P?=?0.001), respectively. In summary, the high level of HOTAIR is intimately associated with an adverse OS in numerous cancers, suggesting that HOTAIR may act as a potential biomarker for the development of malignancies.
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Up-regulation of 91H promotes tumor metastasis and predicts poor prognosis for patients with colorectal cancer.
PLoS ONE
PUBLISHED: 01-01-2014
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Long noncoding RNAs (lncRNAs) play widespread roles in gene regulation and cellular processes. However, the functional roles of lncRNAs in colorectal cancer (CRC) are not yet well elucidated. The aim of the present study was to measure the levels of lncRNA 91H expression in CRC and evaluate its clinical significance and biological roles in the development and progression of CRC.
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Association of the polymorphisms in the Fas/FasL promoter regions with cancer susceptibility: a systematic review and meta-analysis of 52 studies.
PLoS ONE
PUBLISHED: 01-01-2014
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Fas and its ligand (FasL) play an important role in apoptosis and carcinogenesis. Therefore, the potential association of polymorphisms in the Fas (-670A>G, rs1800682; -1377G>A, rs2234767) and FasL (-844C>T, rs763110) with cancer risk has been widely investigated. However, all the currently available results are not always consistent. In this work, we performed a meta-analysis to further determine whether carriers of the polymorphisms in Fas and FasL of interest could confer an altered susceptibility to cancer. All relevant data were retrieved by PubMed and Web of Science, and 52 eligible studies were chosen for this meta-analysis. There was no association of the Fas -670A>G polymorphism with cancer risk in the pooled data. For the Fas -1377G>A and FasL -844C>T polymorphisms, results revealed that the homozygotes of -1377A and -844C were associated with elevated risk of cancer as a whole. Further stratified analysis indicated markedly increased risk for developing breast cancer, gastric cancer, and esophageal cancer, in particular in Asian population. We conclude that carriers of the Fas-1377A and the FasL -844C are more susceptible to the majority of cancers than non-carriers.
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MCPIP1 restricts HIV infection and is rapidly degraded in activated CD4+ T cells.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 11-04-2013
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HIV-1 primarily infects activated CD4+ T cells and macrophages. Quiescent CD4+ T cells, however, possess cellular factors that limit HIV-1 infection at different postentry steps of the viral life cycle. Here, we show that the previously reported immune regulator monocyte chemotactic protein-induced protein 1 (MCPIP1) restricts HIV-1 production in CD4+ T cells. While the ectopic expression of MCPIP1 in cell lines abolished the production of HIV-1, silencing of MCPIP1 enhanced HIV-1 production. Subsequent analysis indicated that MCPIP1 imposes its restriction by decreasing the steady levels of viral mRNA species through its RNase domain. Remarkably, common T-cell stimuli induced the rapid degradation of MCPIP1 in both T-cell lines and quiescent human CD4+ T cells. Lastly, blocking the proteosomal degradation of MCPIP1 by MG132 abrogated HIV-1 production in phorbol 12-myristate 13-acetate/ionomycin-stimulated human CD4+ T cells isolated from healthy donors. Overall, MCPIP1 poses a potent barrier against HIV-1 infection at a posttranscriptional stage. Although the observed HIV restriction conferred by MCPIP1 does not seem to be overcome by any viral protein, it is removed during cellular stimulation. These findings provide insights into the mechanisms of cellular activation-mediated HIV-1 production in CD4+ T cells.
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Conference report: reviving pharmaceutical R&D with translational science, regulatory efficiency and innovative models.
Bioanalysis
PUBLISHED: 10-22-2013
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The 4th Annual Shanghai Symposium on Clinical & Pharmaceutical Solutions through Analysis (CPSA Shanghai 2013) was held on 24-27 April 2013 in Shanghai, China. The meeting provided an educational forum for scientists from pharmaceutical industry, academia, CROs and instrument vendors to share experience and ideas, and discuss current challenges, issues and innovative solutions associated with pharmaceutical R&D. The meeting featured highly interactive events, including diversified symposia, roundtable discussions, workshops, poster sessions and conference awards. Education and specialized training are the foundation of CPSA events. The CPSA Shanghai 2013 meeting also featured an inaugural satellite workshop event in Beijing, as well as joint sessions traditionally held with local bioanalytical and drug metabolism discussion groups.
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Rejuvenation surgery through blepharoplasty incision for mild to moderate upper eyelid sagging in older Asian patients.
J Craniofac Surg
PUBLISHED: 09-17-2013
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The objective of this study was to evaluate the clinical outcome of rejuvenation surgery through double eyelid incision instead of face-lifting for the correction of different types of the upper eyelid sagging in older East Asian patients.
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Poly dimethyl diallyl ammonium coated CMK-5 for sustained oral drug release.
Int J Pharm
PUBLISHED: 08-25-2013
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A new oral sustained drug delivery system (DDS) involving a combination of inorganic mesoporous material (CMK-5) and organic polymer poly dimethyl diallyl ammonium (PDDA) was established to determine its general suitability for use with poorly water soluble drugs. Nimodipine, carvedilol and fenofibrate, three different drugs with acidic or alkaline properties, were selected as model drugs and loaded into carriers. The physicochemical properties of the drug carriers were systematically studied using scanning electron microscopy (SEM), transmission electron microscopy (TEM), and nitrogen adsorption. The structural body changes of the composites in release medium, with or without additional salts, were also studied using particle sizing systems, nitrogen adsorption and zeta potential measurement in order to investigate the sustained release mechanism of the drugs. The results obtained showed that sustained release of drug from the designed DDS was mainly due to the blockage effect arising from the strong swelling of the coated polymers when in contact with release medium. Additional salts, when they reached a certain level, allowed a dramatic burst release. We believe that our designed sustained DDS provide a new option for water insoluble drugs and can be considered as fundamental for those more sophisticated DDS increasingly required in modern medical treatments.
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All-trans retinoic acid prevents epidural fibrosis through NF-?B signaling pathway in post-laminectomy rats.
Neuropharmacology
PUBLISHED: 08-23-2013
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Laminectomy is a widely accepted treatment for lumbar disorders, and epidural fibrosis (EF) is a common complication. EF is thought to cause post-operative pain recurrence after laminectomy or discectomy. All-trans retinoic acid (ATRA) has shown anti-fibrotic, anti-inflammatory, and anti-proliferative functions. The object of this study was to investigate the effects of ATRA on the prevention of EF in post-laminectomy rats. In vitro, the anti-fibrotic effect of ATRA was demonstrated with cultured fibroblasts count, which comprised of those that were cultured with/without ATRA. In vivo, rats underwent laminectomy at the L1-L2 levels. We first demonstrated the beneficial effects using 0.05% ATRA compared to vehicle (control group). We found that a higher concentration of ATRA (0.1%) achieved dose-dependent results. Hydroxyproline content, Rydell score, vimentin-positive cell density, fibroblast density, inflammatory cell density and inflammatory factor expression levels all suggested better outcomes in the 0.1% ATRA rats compared to the other three groups. Presumably, these effects involved ATRAs ability to suppress transforming growth factor (TGF-?1) and interleukin (IL)-6 which was confirmed with reverse-transcriptase polymerase chain reaction (RT-PCR). Finally we demonstrated that ATRA down-regulated nuclear factor (NF)-?B by immunohistochemistry and western blotting for p65 and inhibition of ?B (I?B?), respectively. Our findings indicate that topical application of ATRA can inhibit fibroblast proliferation, decrease TGF-?1 and IL-6 expression level, and prevent epidural scar adhesion in rats. The highest concentration employed in this study (0.1%) was the most effective. ATRA suppressed EF through down-regulating NF-?B signaling, whose specific mechanism is suppression of I?B phosphorylation and proteolytic degradation.
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Onset of atonal expression in Drosophila retinal progenitors involves redundant and synergistic contributions of Ey/Pax6 and So binding sites within two distant enhancers.
Dev. Biol.
PUBLISHED: 08-12-2013
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Proneural transcription factors drive the generation of specialized neurons during nervous system development, and their dynamic expression pattern is critical to their function. The activation of the proneural gene atonal (ato) in the Drosophila eye disc epithelium represents a critical step in the transition from retinal progenitor cell to developing photoreceptor neuron. We show here that the onset of ato transcription depends on two distant enhancers that function differently in subsets of retinal progenitor cells. A detailed analysis of the crosstalk between these enhancers identifies a critical role for three binding sites for the Retinal Determination factors Eyeless (Ey) and Sine oculis (So). We show how these sites interact to induce ato expression in distinct regions of the eye field and confirm them to be occupied by endogenous Ey and So proteins in vivo. Our study suggests that Ey and So operate differently through the same 3 cis-regulatory sites in distinct populations of retinal progenitors.
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[Effect of aneurysm clipping on hemorrhage volume in the subarachnoid space].
Nan Fang Yi Ke Da Xue Xue Bao
PUBLISHED: 07-31-2013
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To evaluate the effect of aneurysm clipping and partial blood clot removal in the subarachnoid space on hemorrhage volume in the subarachnoid space and cerebral vasospasm in patients with different Fisher grades.
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Downregulation of CD147 expression by RNA interference inhibits HT29 cell proliferation, invasion and tumorigenicity in vitro and in vivo.
Int. J. Oncol.
PUBLISHED: 07-18-2013
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We investigated the effect of CD147 silencing on HT29 cell proliferation and invasion. We constructed a novel short hairpin RNA (shRNA) expression vector pYr-mir30-shRNA. The plasmid was transferred to HT29 cells. The expression of CD147, MCT1 (lactate transporters monocarboxylate transporter 1) and MCT4 (lactate transporters monocarboxylate transporter 4) were monitored by quantitative PCR and western blotting, respectively. The MMP-2 (matrix metalloproteinase-2) and MMP-9 (matrix metalloproteinase-9) activities were determined by gelatin zymography assay, while the intracellular lactate concentration was determined by the lactic acid assay kit. WST-8 assay was used to determine the HT29 cell proliferation and the chemosensitivity. Invasion assay was used to determine the invasion of HT29 cells. In addition, we established a colorectal cancer model, and detected CD147 expression in vivo. The results showed that the expression of CD147 and MCT1 was significantly reduced at both mRNA and protein levels, and also the activity of MMP-2 and MMP-9 was reduced. The proliferation and invasion were decreased, but chemosensitivity to cisplatin was increased. In vivo, the CD147 expression was also significantly decreased, and reduced the tumor growth after CD147 gene silencing. The results demonstrated that silencing of CD147 expression inhibited the proliferation and invasion, suggesting CD147 silencing might be an adjuvant gene therapy strategy to chemotherapy.
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ERK1/2 signalling pathway is involved in CD147-mediated gastric cancer cell line SGC7901 proliferation and invasion.
Exp. Biol. Med. (Maywood)
PUBLISHED: 07-04-2013
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This study aimed to investigate the role of CD147 in the progression of gastric cancer and the signalling pathway involved in CD147-mediated gastric cancer cell line SGC7901 proliferation and invasion. Short hairpin RNA (shRNA) expression vectors targeting CD147 were constructed to silence CD147, and the expression of CD147 was monitored by quantitative realtime reverse transcriptase polymerase chain reaction and Western blot and further confirmed by immunohistochemistry in vivo. Cell proliferation was determined by Cell Counting Kit-8 assay, the activities of matrix metalloproteinase (MMP)-2 and MMP-9 were determined by gelatin zymography, and the invasion of SGC7901 was determined by invasion assay. The phosphorylation and non-phosphorylation of the mitogen-activated protein kinases, extracellular signal-regulated kinase1/2 (ERK1/2), P38 and c-Jun NH2-terminal kinase were examined by Western blot. Additionally, the ERK1/2 inhibitor U0126 were used to confirm the signalling pathway involved in CD147-mediated SGC7901 progression. The BALB/c nude mice were used to study tumour progression in vivo. The results revealed that CD147 silencing inhibited the proliferation and invasion of SGC7901 cells, and down-regulated the activities of MMP-2 and MMP-9 and the phosphorylation of the ERK1/2 in SGC7901 cells. ERK1/2 inhibitor U0126 decreased the proliferation, and invasion of SGC7901 cells, and down-regulated the MMP-2 and MMP-9 activities. In a nude mouse model of subcutaneous xenografts, the tumour volume was significantly smaller in the SGC7901/shRNA group compared to the SGC7901 and SGC7901/snc-RNA group. Immunohistochemistry analysis showed that CD147 and p-ERK1/2 protein expressions were down-regulated in the SGC7901/shRNA2 group compared to the SGC7901 and SGC7901/snc-RNA group. These results suggest that ERK1/2 pathway involves in CD147-mediated gastric cancer growth and invasion. These findings further highlight the importance of CD147 in cancer progression, indicating that CD147 would be an attractive therapeutic target for gastric cancer.
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Human placental trophoblasts confer viral resistance to recipient cells.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 07-01-2013
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Placental trophoblasts form the interface between the fetal and maternal environments and serve to limit the maternal-fetal spread of viruses. Here we show that cultured primary human placental trophoblasts are highly resistant to infection by a number of viruses and, importantly, confer this resistance to nonplacental recipient cells by exosome-mediated delivery of specific microRNAs (miRNAs). We show that miRNA members of the chromosome 19 miRNA cluster, which are almost exclusively expressed in the human placenta, are packaged within trophoblast-derived exosomes and attenuate viral replication in recipient cells by the induction of autophagy. Together, our findings identify an unprecedented paracrine and/or systemic function of placental trophoblasts that uses exosome-mediated transfer of a unique set of placental-specific effector miRNAs to directly communicate with placental or maternal target cells and regulate their immunity to viral infections.
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Clinical diagnosis and treatment of alpha-fetoprotein-negative small hepatic lesions.
Chin. J. Cancer Res.
PUBLISHED: 06-27-2013
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We examined 103 cases over the last five years and discussed diagnosis and treatment of alpha-fetoprotein (AFP)-negative small hepatic lesions.
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Nanog, a novel prognostic marker for lung cancer.
Surg Oncol
PUBLISHED: 06-26-2013
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To investigate the expression of the stem cell marker Nanog in lung cancer tissues and the correlations between Nanog expression and clinic-pathologic characteristics as well as prognosis of lung cancer.
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Structural basis for interaction between Mycobacterium smegmatis Ms6564, a TetR family master regulator, and its target DNA.
J. Biol. Chem.
PUBLISHED: 06-26-2013
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Master regulators, which broadly affect expression of diverse genes, play critical roles in bacterial growth and environmental adaptation. However, the underlying mechanism by which such regulators interact with their cognate DNA remains to be elucidated. In this study, we solved the crystal structure of a broad regulator Ms6564 in Mycobacterium smegmatis and its protein-operator complex at resolutions of 1.9 and 2.5 ?, respectively. Similar to other typical TetR family regulators, two dimeric Ms6564 molecules were found to bind to opposite sides of target DNA. However, the recognition helix of Ms6564 inserted only slightly into the DNA major groove. Unexpectedly, 11 disordered water molecules bridged the interface of TetR family regulator DNA. Although the DNA was deformed upon Ms6564 binding, it still retained the conformation of B-form DNA. Within the DNA-binding domain of Ms6564, only two amino acids residues directly interacted with the bases of cognate DNA. Lys-47 was found to be essential for the specific DNA binding ability of Ms6564. These data indicate that Ms6564 can bind DNA with strong affinity but makes flexible contacts with DNA. Our study suggests that Ms6564 might slide more easily along the genomic DNA and extensively regulate the expression of diverse genes in M. smegmatis.
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Lipoic acid restores age-associated impairment of brain energy metabolism through the modulation of Akt/JNK signaling and PGC1? transcriptional pathway.
Aging Cell
PUBLISHED: 06-23-2013
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This study examines the progress of a hypometabolic state inherent in brain aging with an animal model consisting of Fischer 344 rats of young, middle, and old ages. Dynamic microPET scanning demonstrated a significant decline in brain glucose uptake at old ages, which was associated with a decrease in the expression of insulin-sensitive neuronal glucose transporters GLUT3/4 and of microvascular endothelium GLUT1. Brain aging was associated with an imbalance between the PI3K/Akt pathway of insulin signaling and c-Jun N-terminal kinase (JNK) signaling and a downregulation of the PGC1?-mediated transcriptional pathway of mitochondrial biogenesis that impinged on multiple aspects of energy homeostasis. R-(+)-lipoic acid treatment increased glucose uptake, restored the balance of Akt/JNK signaling, and enhanced mitochondrial bioenergetics and the PGC1?-driven mitochondrial biogenesis. It may be surmised that impairment of a mitochondria-cytosol-nucleus communication is underlying the progression of the age-related hypometabolic state in brain; the effects of lipoic acid are not organelle-limited, but reside on the functional and effective coordination of this communication that results in improved energy metabolism.
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Is resilience only skin deep?: rural african americans socioeconomic status-related risk and competence in preadolescence and psychological adjustment and allostatic load at age 19.
Psychol Sci
PUBLISHED: 05-30-2013
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Many African American youth may develop high levels of allostatic load, a measure of physiological wear and tear on the body, by developing psychosocial competence under conditions of high risk related to socioeconomic status (SES). The current study was designed to test this hypothesis, which is based on John Henryism theory. In a representative sample of 489 African American youth living in the rural South, cumulative SES-related risks and teacher-reported competence were assessed at ages 11 to 13; depressive symptoms, externalizing behavior, and allostatic load were assessed at age 19. The data revealed that rural African American preadolescents who evinced high psychosocial competence under conditions of high cumulative SES-related risk displayed low levels of adjustment problems along with high allostatic load at age 19. These results suggest that, for many rural African Americans, resilience may indeed be only "skin deep."
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Suppression of KIF3B Expression Inhibits Human Hepatocellular Carcinoma Proliferation.
Dig. Dis. Sci.
PUBLISHED: 05-21-2013
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Human hepatocellular carcinoma (HCC) is one of the most common fatal cancers and an important health problem worldwide, but its mechanism is still unclear. Microtubule (MT) kinesin motor proteins orchestrate a variety of cellular processes (e.g. mitosis, motility and organelle transportation) and have been involved in human carcinogenesis. KIF3B, the kinesin superfamily of proteins (KIFs), plays an important role in the regulation of mitotic progression.
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Harsh Parenting and Adolescent Health: A Longitudinal Analysis With Genetic Moderation.
Health Psychol
PUBLISHED: 05-13-2013
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Objective: This study was designed to examine the prospective relations of harsh parenting during preadolescence, anger across adolescence, and a health phenotype at late adolescence among African American youths living in the rural South. A second purpose was to determine whether, for genetic reasons, some youths will be more sensitive than others to a harsh parenting to anger to poor health pathway. Method: Participants were 368 youths (age 11.2 at the first assessment) who provided data on receipt of harsh parenting during preadolescence (ages 11 to 13), anger across adolescence (ages 16 to 18), and a health phenotype consisting of C Reactive Protein, depressive symptoms, and health problems at age 19. Youths were genotyped at the 5-HTTLPR at age 16. Results: The data analysis revealed that (a) harsher parenting was associated positively across time with anger and poor health, (b) anger across adolescence also was associated positively across time with poor health, (c) anger served as a mediator connecting harsh parenting and poor health, and (d) the harsh parenting to anger to poor health pathway was significant only for youths carrying one or two copies of a short allele at the 5-HTTLPR. Conclusions: These findings are consistent with the hypothesis that harsh parent-child interactions presage health through effects on emotion regulation, particularly anger. This mediational pathway pertained only to youths carrying a gene that confers sensitivity and reactivity to harsh family processes and the negative emotional states they occasion. (PsycINFO Database Record (c) 2013 APA, all rights reserved).
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Toll-like receptor 4 increases intestinal permeability through up-regulation of membrane PKC activity in alcoholic steatohepatitis.
Alcohol
PUBLISHED: 05-09-2013
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Intestinal hyperpermeability is a causal factor for the development of alcoholic endotoxemia and steatohepatitis. However, the mechanisms governing this link remain unknown. The purpose of this study was to determine whether toll-like receptor 4 (TLR4) is involved in ethanols deleterious effects on the intestinal barrier. Caco-2 cells were incubated in vitro with 1-10% ethanol. The results indicated that ethanol had a dose-dependent effect in increasing TLR4 expression and intercellular permeability. Then the effects of TLR4 on protein kinase C (PKC) and the intercellular junction protein occludin were assessed with and without pretreatment with a TLR4 inhibitor. The results indicated that TLR4 increased nonspecific PKC activity and reduced the expression of phosphorylated occludin in the membrane, which increased intercellular permeability. These effects were prevented by pretreatment with TLR4 mAb. Wild-type C57BL/6 mice were fed an ethanol or isocaloric liquid diet for 6 weeks. Hepatitis was diagnosed by the presence of an associated elevated blood endotoxin level. Chronic ethanol treatment significantly elevated blood endotoxin levels, intestinal permeability, and the expression of TLR4 in the ileum and colon. Moreover, ethanol exposure reduced the distribution of phosphorylated occludin in the intestinal epithelium because of PKC activation. In conclusion, chronic ethanol exposure induces a high response of TLR4 to lipopolysaccharide (LPS), and TLR4 increases intestinal permeability through down-regulation of phosphorylated occludin expression in the intestinal epithelial barrier, accompanied by membrane PKC hyperactivity.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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