JoVE Visualize What is visualize?
Stop Reading. Start Watching.
Advanced Search
Stop Reading. Start Watching.
Regular Search
Find video protocols related to scientific articles indexed in Pubmed.
Developing sustainable global health technologies: Insight from an initiative to address neonatal hypothermia.
J Public Health Policy
PUBLISHED: 10-31-2014
Show Abstract
Hide Abstract
Relative to drugs, diagnostics, and vaccines, efforts to develop other global health technologies, such as medical devices, are limited and often focus on the short-term goal of prototype development instead of the long-term goal of a sustainable business model. To develop a medical device to address neonatal hypothermia for use in resource-limited settings, we turned to principles of design theory: (1) define the problem with consideration of appropriate integration into relevant health policies, (2) identify the users of the technology and the scenarios in which the technology would be used, and (3) use a highly iterative product design and development process that incorporates the perspective of the user of the technology at the outset and addresses scalability. In contrast to our initial idea, to create a single device, the process guided us to create two separate devices, both strikingly different from current solutions. We offer insights from our initial experience that may be helpful to others engaging in global health technology development.Journal of Public Health Policy advance online publication, 13 November 2014; doi:10.1057/jphp.2014.44.
Related JoVE Video
Mechanoregulation of osteoblast-like MG-63 cell activities by cyclic stretching.
J. Formos. Med. Assoc.
PUBLISHED: 06-26-2014
Show Abstract
Hide Abstract
Mechanical loading plays an important role in regulating bone formation and remodeling. Relevant mechanical stretching can increase the proliferation and differentiation of osteoblastic cells in vitro. However, little is known about the effects of supraphysiological high-level mechanical stretching on the growth and cell cycle progression of osteoblastic cells.
Related JoVE Video
Reduced structural integrity and functional lateralization of the dorsal language pathway correlate with hallucinations in schizophrenia: A combined diffusion spectrum imaging and functional magnetic resonance imaging study.
Psychiatry Res
PUBLISHED: 06-15-2014
Show Abstract
Hide Abstract
Recent studies suggest that structural and functional alterations of the language network are associated with auditory verbal hallucinations (AVHs) in schizophrenia. However, the ways in which the underlying structure and function of the network are altered and how these alterations are related to each other remain unclear. To elucidate this, we used diffusion spectrum imaging (DSI) to reconstruct the dorsal and ventral pathways and employed functional magnetic resonance imaging (fMRI) in a semantic task to obtain information about the functional activation in the corresponding regions in 18 patients with schizophrenia and 18 matched controls. The results demonstrated decreased structural integrity in the left ventral, right ventral and right dorsal tracts, and decreased functional lateralization of the dorsal pathway in schizophrenia. There was a positive correlation between the microstructural integrity of the right dorsal pathway and the functional lateralization of the dorsal pathway in patients with schizophrenia. Additionally, both functional lateralization of the dorsal pathway and microstructural integrity of the right dorsal pathway were negatively correlated with the scores of the delusion/hallucination symptom dimension. Our results suggest that impaired structural integrity of the right dorsal pathway is related to the reduction of functional lateralization of the dorsal pathway, and these alterations may aggravate AVHs in schizophrenia.
Related JoVE Video
Revisiting the stability of mini-implants used for orthodontic anchorage.
J. Formos. Med. Assoc.
PUBLISHED: 06-08-2014
Show Abstract
Hide Abstract
The aim of this study is to comprehensively analyze the potential factors affecting the failure rates of three types of mini-implants used for orthodontic anchorage.
Related JoVE Video
Differential Synaptic and Extrasynaptic Glutamate-Receptor Alterations in Striatal Medium-Sized Spiny Neurons of Aged YAC128 Huntington's Disease Mice.
PLoS Curr
PUBLISHED: 06-05-2014
Show Abstract
Hide Abstract
Huntington's disease (HD) is a late-onset, slowly progressing neurodegenerative disorder caused by an expansion of glutamine repeats. The YAC128 mouse model has been widely used to study the progression of HD symptoms, but little is known about synaptic alterations in very old animals. The present experiments examined synaptic properties of striatal medium-sized spiny neurons (MSNs) in 16 month-old YAC128 mice. These mice were crossed with mice expressing enhanced green fluorescent protein (EGFP) under the control of either D1 or D2 dopamine receptor promoters to identify MSNs originating the direct and indirect pathways, respectively. The input-output curves of evoked excitatory postsynaptic currents mediated by activation of the ?-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) or N-methyl-D-aspartate (NMDA) receptors were reduced in MSNs in both pathways. In the presence of DL-threo-?-Benzyloxyaspartic acid (DL-TBOA), a glutamate transporter blocker used to increase activation of extrasynaptic receptors, NMDA receptor-mediated currents displayed altered amplitudes, longer decay times, and greater charge (response areas) in both direct and indirect pathway MSNs in YAC128 mice compared to wildtype controls. Amplitudes were significantly increased, primarily in direct pathway MSNs while normalized areas were significantly increased only in indirect pathway MSNs, suggesting that the two types of MSNs are affected in different ways. It may be that indirect pathway neurons are more susceptible to changes in glutamate transport. Taken together, the present findings demonstrate differential alterations in synaptic versus extrasynaptic NMDA receptors in both direct and indirect pathway MSNs in late HD, which may contribute to the dysfunction and degeneration in both pathways.
Related JoVE Video
Thoracic amyloidomas: Two case reports of an evasive diagnosis.
JRSM Open
PUBLISHED: 06-01-2014
Show Abstract
Hide Abstract
Amyloidosis is a rare differential diagnosis of a mass detected in the chest. Amyloidoma is caused by a local proliferation of clonal B-cells secreting an unstable immunoglobulin light chain which accumulates. FDG-PET scan are useful but not specific. Treatment is generally by local resection for treatment of symptoms. We report two cases of amyloidomas, which are rare entities characterised by large local amyloid deposits. These can occur in the upper respiratory tract, soft tissues and central nervous system.(1.)
Related JoVE Video
Translational control by heme-regulated eIF2? kinase during erythropoiesis.
Curr. Opin. Hematol.
PUBLISHED: 04-10-2014
Show Abstract
Hide Abstract
This review will provide an overview of the translational regulation of globin mRNAs and integrated stress response (ISR) during erythropoiesis by heme-regulated eIF2? kinase (HRI). HRI is an intracellular heme sensor that coordinates heme and globin synthesis in erythropoiesis by inhibiting protein synthesis of globins and heme biosynthetic enzymes during heme deficiency.
Related JoVE Video
Health behaviors and needs of melanoma survivors.
Support Care Cancer
PUBLISHED: 01-30-2014
Show Abstract
Hide Abstract
Little is known about melanoma survivors' long-term symptoms, sun protection practices, and support needs from health providers.
Related JoVE Video
Attention deficit hyperactivity disorder and N-methyl-D-aspartate (NMDA) dysregulation.
Curr. Pharm. Des.
PUBLISHED: 01-09-2014
Show Abstract
Hide Abstract
Attention deficit hyperactivity disorder (ADHD) is a long recognized and common childhood disorder. ADHD adolescents tend to encounter more difficulties in school and peer relationships, whereas ADHD adults have more occupational and interpersonal difficulties. However, with the treatment of central nervous system (CNS) stimulants, 10-20 % of the patients still remain poor responders to treatment. Among hypotheses for ADHD, dysfunction of N-methyl-D-aspartate (NMDA)-type glutamate receptors has recently been suggested by accumulating genetic and animal studies. This article systemically reviews evidence supporting NMDA dysfunction as a potential ADHD pathogenesis from perspectives of neurodevelopment, attentional circuitry, and impulse inhibition. The review also addresses the development of novel treatments for ADHD via modulation of glutamatergic system, particularly the NMDA/glycine site. These so-called NMDA enhancers may provide a new treatment option for patients with ADHD.
Related JoVE Video
Omega-3 fatty acids in the prevention of interferon-alpha-induced depression: results from a randomized, controlled trial.
Biol. Psychiatry
PUBLISHED: 01-06-2014
Show Abstract
Hide Abstract
Interferon (IFN)-? therapy for chronic hepatitis C virus infection is frequently associated with depression. The routine prophylaxis with antidepressants might expose patients to adverse effects, hence, the need for alternative preventive interventions. Omega-3 polyunsaturated fatty acids are safe and effective essential nutritional compounds used for the treatment of depression, putatively through an anti-inflammatory action. In addition, lower erythrocyte levels of omega-3 polyunsaturated fatty acids have been associated with an increased risk of IFN-induced depression.
Related JoVE Video
Areca nut components affect COX-2, cyclin B1/cdc25C and keratin expression, PGE2 production in keratinocyte is related to reactive oxygen species, CYP1A1, Src, EGFR and Ras signaling.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Chewing of betel quid (BQ) increases the risk of oral cancer and oral submucous fibrosis (OSF), possibly by BQ-induced toxicity and induction of inflammatory response in oral mucosa.
Related JoVE Video
TCDD promotes lung tumors via attenuation of apoptosis through activation of the Akt and ERK1/2 signaling pathways.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a multiple-site, multiple-species carcinogen that induces cancer in multiple organs. The molecular mechanisms underlying TCDD-induced lung tumorigenesis remain unclear. In the present study, a two-stage lung tumorigenesis model was established by administrating a single low dose of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) combined with TCDD to female A/J mice. The results indicated that TCDD combined with low-dose NNK has a significant tumor-promoting effect compared with TCDD or low-dose NNK alone. Resistance to apoptosis is a hallmark of cancer and is thought to be one of the tumor-promoting mechanisms regulated by TCDD. We performed an additional series of experiments in the normal human bronchial epithelial cell line Beas2B cells, in which TCDD was combined with the apoptosis inducer staurosporine. Our in vitro results confirmed that TCDD could rescue cells from apoptosis induced by staurosporine. The inhibition of apoptosis is likely mediated by the activation of the Akt and ERK1/2 pathways, as determined by the effectiveness of pathway-specific inhibitors in abrogating the anti-apoptotic activity of TCDD. In conclusion, we demonstrated that TCDD promoted NNK-induced lung tumorigenesis and revealed that TCDD inhibits staurosporine-induced apoptosis, at least in part, through the Akt and ERK1/2 signaling pathways.
Related JoVE Video
The regulation of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced lung tumor promotion by estradiol in female A/J mice.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Epidemiological studies indicate that women are at a higher risk developing lung cancer than men are. It is suggested that estrogen is one of the most important factors in lung cancer development in females. Additionally, cigarette smoke, and environmental pollutants, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), may play salient roles in female lung carcinogenesis. However, the mechanisms responsible for the interaction of these factors in the promotion of lung cancer are still poorly understood. The present study was designed to explore two ideas: first, the synergistic lung tumorigenic effects of 4-(methylnitrosamino)-1-(3-pyridyl)-butanol (NNK) combined with TCDD, 17?-estradiol (E2) or both through a long-term treatment experiment, and second, to identify early changes in the inflammatory and signaling pathways through short-term treatment experiments. The results indicate that A/J mice given E2 had strong effects in potentiating NNK-induced activation of MAPK signaling, NF?B, and COX-2 expression. In the long-term exposure model, E2 had a strong tumor promoting effect, whereas TCDD antagonized this effect in A/J mice. We conclude that treatment with NNK combined with either E2 or TCDD induces lung carcinogenesis and the promotion effects could be correlated with lung inflammation. E2 was shown to potentiate NNK-induced inflammation, cell proliferation, thereby leading to lung tumorigenesis.
Related JoVE Video
The pentachlorophenol metabolite tetrachlorohydroquinone induces massive ROS and prolonged p-ERK expression in splenocytes, leading to inhibition of apoptosis and necrotic cell death.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Pentachlorophenol (PCP) has been used extensively as a biocide and a wood preservative and has been reported to be immunosuppressive in rodents and humans. Tetrachlorohydroquinone (TCHQ) is a major metabolite of PCP. TCHQ has been identified as the main cause of PCP-induced genotoxicity due to reactive oxidant stress (ROS). However, the precise mechanisms associated with the immunotoxic effects of PCP and TCHQ remain unclear. The aim of this study was to examine the effects of PCP and TCHQ on the induction of ROS and injury to primary mouse splenocytes. Our results shown that TCHQ was more toxic than PCP and that a high dose of TCHQ led to necrotic cell death of the splenocytes through induction of massive and sudden ROS and prolonged ROS-triggered ERK activation. Inhibition of ROS production by N-acetyl-cysteine (NAC) partially restored the mitochondrial membrane potential, inhibited ERK activity, elevated caspase-3 activity and PARP cleavage, and, eventually, switched the TCHQ-induced necrosis to apoptosis. We suggest that prolonged ERK activation is essential for TCHQ-induced necrosis, and that ROS play a pivotal role in the different TCHQ-induced cell death mechanisms.
Related JoVE Video
Establishing a mobile environmental survey system for real-time emergency response.
Health Phys
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
After the 9/11 incident, the needs of environmental survey mobility were increased because the time and place of terrorism attacks are more unpredictable than traditional radiological and nuclear accident. In recent years, more and more attention has been paid to the radiation protection of the survey workers in the field. The Institute of Nuclear Energy Research has established a Mobile Environmental Survey System (MESS) for real time emergency response by using network communication. The system consists of three major functional components: the mobile units, the management center, and the work stations. The system can display real time survey data on an electric map collected by the survey workers from the remote field to improve the environmental survey mobility. The system can also send the survey workers messages/warnings according to geographic information and the location of the mobile unit to keep them from unnecessary radiation exposure that may be an effective way to the goal of "as low as reasonably achievable" for environmental surveys. MESS has been adopted as an essential tool for emergency response in Taiwan. The application experience of MESS in the relative exercises will be discussed in this paper.
Related JoVE Video
Speech-induced atrial tachycardia: an unusual presentation of supraventricular tachycardia.
J. Cardiovasc. Electrophysiol.
PUBLISHED: 07-21-2013
Show Abstract
Hide Abstract
A 63-year-old male radio announcer was admitted with a narrow complex, long RP tachycardia. While in the awake state, the patient spoke in his radio voice, initiating and maintaining the tachycardia. Three-dimensional electroanatomic mapping during electrophysiology study localized the tachycardia to the ostium of the right superior pulmonary vein. After single radiofrequency energy application, no further arrhythmias were inducible with speech. At more than 1 year of follow-up, the patient had no recurrences and continues to work as a radio announcer.
Related JoVE Video
Opioid self-administration results in cell-type specific adaptations of striatal medium spiny neurons.
Behav. Brain Res.
PUBLISHED: 06-25-2013
Show Abstract
Hide Abstract
Medium-sized spiny neurons (MSNs), the predominant neuronal population of the striatum, are an integral component of the many cortical and limbic pathways associated with reward-related behaviors. A differential role of the D1 receptor-enriched (D1) MSNs of the striatonigral direct pathway, as compared with the D2 receptor-enriched (D2) MSNs of the striatopallidal indirect pathway, in mediating the addictive behaviors associated with cocaine is beginning to emerge. However, whether opioids, well-known analgesics with euphoric properties, similarly induce dissociable signaling adaptations in these neurons remains unclear. Transgenic mice expressing green fluorescent protein (GFP)-labeled D1 or D2 neurons were implanted with intravenous jugular catheters. Mice learned to self-administer 0.1mg/kg/infusion of the opioid remifentanil during 2h sessions over 13 contiguous days. Thereafter, the electrophysiological properties of D1- and D2-MSNs in the shell region of the nucleus accumbens (NAc) were assessed. We found that prior opioid exposure did not alter the basic membrane properties nor the kinetics or amplitude of miniature excitatory postsynaptic currents (mEPSCs). However, when challenged with the mu opioid receptor (?OR) agonist DAMGO, the characteristic inhibitory profile of this receptor was altered. DAMGO inhibited the frequency of mEPSCs in D1-MSNs from control mice receiving saline and in D2-MSNs from mice exposed to remifentanil or saline, but this inhibitory profile was reduced in D1-MSNs from mice receiving remifentanil. Remifentanil exposure also altered the probability of glutamate release onto D1-, but not D2-MSNs. Together these results suggest a D1-pathway specific effect associated with the acquisition of opioid-seeking behaviors.
Related JoVE Video
Graphene oxide induces toll-like receptor 4 (TLR4)-dependent necrosis in macrophages.
ACS Nano
PUBLISHED: 06-11-2013
Show Abstract
Hide Abstract
Graphene and graphene-based nanomaterials display novel and beneficial chemical, electrical, mechanical, and optical characteristics, which endow these nanomaterials with promising applications in a wide spectrum of areas such as electronics and biomedicine. However, its toxicity on health remains unknown and is of great concern. In the present study, we demonstrated that graphene oxide (GO) induced necrotic cell death to macrophages. This toxicity is mediated by activation of toll-like receptor 4 (TLR4) signaling and subsequently in part via autocrine TNF-? production. Inhibition of TLR4 signaling with a selective inhibitor prevented cell death nearly completely. Furthermore, TLR4-deficient bone marrow-derived macrophages were resistant to GO-triggered necrosis. Similarly, GO did not induce necrosis of HEK293T/TLR4-null cells. Macrophagic cell death upon GO treatment was partially attributed to RIP1-RIP3 complex-mediated programmed necrosis downstream of TNF-? induction. Additionally, upon uptake into macrophages, GO accumulated primarily in cytoplasm causing dramatic morphologic alterations and a significant reduction of the macrophagic ability in phagocytosis. However, macrophagic uptake of GO may not be required for induction of necrosis. GO exposure also caused a large increase of intracellular reactive oxygen species (ROS), which contributed to the cause of cell death. The combined data reveal that interaction of GO with TLR4 is the predominant molecular mechanism underlying GO-induced macrophagic necrosis; also, cytoskeletal damage and oxidative stress contribute to decreased viability and function of macrophages upon GO treatment.
Related JoVE Video
An integrin binding-defective mutant of insulin-like growth factor-1 (R36E/R37E IGF1) acts as a dominant-negative antagonist of the IGF1 receptor (IGF1R) and suppresses tumorigenesis but still binds to IGF1R.
J. Biol. Chem.
PUBLISHED: 05-21-2013
Show Abstract
Hide Abstract
Insulin-like growth factor-1 (IGF1) is a major therapeutic target for cancer. We recently reported that IGF1 directly binds to integrins (?v?3 and ?6?4) and induces ternary complex formation (integrin-IGF1-IGF1 receptor (IGF1R)) and that the integrin binding-defective mutant of IGF1 (R36E/R37E) is defective in signaling and ternary complex formation. These findings predict that R36E/R37E competes with WT IGF1 for binding to IGF1R and inhibits IGF signaling. Here, we described that excess R36E/R37E suppressed cell viability increased by WT IGF1 in vitro in non-transformed cells. We studied the effect of R36E/R37E on viability and tumorigenesis in cancer cell lines. We did not detect an effect of WT IGF1 or R36E/R37E in cancer cells under anchorage-dependent conditions. However, under anchorage-independent conditions, WT IGF1 enhanced cell viability and induced signals, whereas R36E/R37E did not. Notably, excess R36E/R37E suppressed cell viability and signaling induced by WT IGF1 under anchorage-independent conditions. Using cancer cells stably expressing WT IGF1 or R36E/R37E, we determined that R36E/R37E suppressed tumorigenesis in vivo, whereas WT IGF1 markedly enhanced it. R36E/R37E suppressed the binding of WT IGF1 to the cell surface and the subsequent ternary complex formation induced by WT IGF1. R36E/R37E suppressed activation of IGF1R by insulin. WT IGF1, but not R36E/R37E, induced ternary complex formation with the IGF1R/insulin receptor hybrid. These findings suggest that 1) IGF1 induces signals under anchorage-independent conditions and that 2) R36E/R37E acts as a dominant-negative inhibitor of IGF1R (IGF1 decoy). Our results are consistent with a model in which ternary complex formation is critical for IGF signaling.
Related JoVE Video
Pacemaker GABA synaptic activity may contribute to network synchronization in pediatric cortical dysplasia.
Neurobiol. Dis.
PUBLISHED: 05-14-2013
Show Abstract
Hide Abstract
Spontaneous pacemaker ?-aminobutyric acid (GABA) receptor-mediated synaptic activity (PGA) occurs in a subset of tissue samples from pediatric epilepsy surgery patients. In the present study, based on single-cell electrophysiological recordings from 120 cases, we describe the etiologies, cell types, and primary electrophysiological features of PGA. Cells displaying PGA occurred more frequently in the areas of greatest anatomical abnormality in cases of focal cortical dysplasia (CD), often associated with hemimegalencephaly (HME), and only rarely in non-CD etiologies. PGA was characterized by rhythmic synaptic events (5-10Hz) and was observed in normal-like, dysmorphic cytomegalic, and immature pyramidal neurons. PGA was action potential-dependent, mediated by GABAA receptors, and unaffected by antagonism of glutamate receptors. We propose that PGA is a unique electrophysiological characteristic associated with CD and HME. It could represent an abnormal signal that may contribute to epileptogenesis in malformed postnatal cortex by facilitating pyramidal neuron synchrony.
Related JoVE Video
The immunotoxic effects of dual exposure to PCP and TCDD.
Chem. Biol. Interact.
PUBLISHED: 05-07-2013
Show Abstract
Hide Abstract
Pentachlorophenol (PCP) was a commonly used fungicide, herbicide, insecticide, and bactericide in industrial, agricultural, and domestic settings; however, it was also contaminated with polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs). It has been reported that technical grade PCP had immunosuppressive effects and that the immune system was the major target of PCDD/PCDFs toxicity. Although the immune response after exposure to PCP or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been studied, the toxic effects of exposure to both PCP and TCDD have not yet been reported. The aim of this study was to evaluate the effects on immune cells from mice intraperitoneally immunized with OVA and subsequently treated with PCP or TCDD alone or in combination by gavage. The animals were terminated on day 7 and 14, and the spleen and plasma samples were collected for immunotoxicity evaluation. The numbers and populations of splenocytes, T cell-derived cytokines produced by splenocytes, splenocyte-generated cytotoxicity and OVA-specific antibodies in plasma were investigated. Our results indicate that the spleen/body weight ratio and splenocyte number was reduced by TCDD alone; in addition, this reduction was enhanced when TCDD was combined with PCP. Exposure to TCDD alone or in conjunction with PCP suppressed many ovalbumin (OVA)-stimulated cytokines, including IL-2, IFN-?, IL-4, IL-5, and IL-10. Furthermore, the immunoglobulins IgG and IgM were suppressed in mice administered by PCP alone, but the suppressive effects were greater in mice treated with TCDD alone or in combination with PCP. Co-exposure to PCP and TCDD resulted in an antagonistic effect on TCDD-induced suppression of IFN-? and IL-10. Our results demonstrate that PCP alone is immunotoxic, regardless of the presence of TCDD. PCP led to mild changes in cytokine secretion, and it compromised splenocyte-generated cytotoxicity and IgM and IgG antibody production on day 7. The finding that PCP antagonizes TCDD-induced IFN-? suppression could be due to the competitive binding of PCP to AhR (aryl hydrocarbon receptor).
Related JoVE Video
Multiple sources of striatal inhibition are differentially affected in Huntingtons disease mouse models.
J. Neurosci.
PUBLISHED: 04-26-2013
Show Abstract
Hide Abstract
In Huntingtons disease (HD) mouse models, spontaneous inhibitory synaptic activity is enhanced in a subpopulation of medium-sized spiny neurons (MSNs), which could dampen striatal output. We examined the potential source(s) of increased inhibition using electrophysiological and optogenetic methods to assess feedback and feedforward inhibition in two transgenic mouse models of HD. Single whole-cell patch-clamp recordings demonstrated that increased GABA synaptic activity impinges principally on indirect pathway MSNs. Dual patch recordings between MSNs demonstrated reduced connectivity between MSNs in HD mice. However, while connectivity was strictly unidirectional in controls, in HD mice bidirectional connectivity occurred. Other sources of increased GABA activity in MSNs also were identified. Dual patch recordings from fast spiking (FS) interneuron-MSN pairs demonstrated greater but variable amplitude responses in MSNs. In agreement, selective optogenetic stimulation of parvalbumin-expressing, FS interneurons induced significantly larger amplitude MSN responses in HD compared with control mice. While there were no differences in responses of MSNs evoked by activating single persistent low-threshold spiking (PLTS) interneurons in recorded pairs, these interneurons fired more action potentials in both HD models, providing another source for increased frequency of spontaneous GABA synaptic activity in MSNs. Selective optogenetic stimulation of somatostatin-expressing, PLTS interneurons did not reveal any significant differences in responses of MSNs in HD mice. These findings provide strong evidence that both feedforward and to a lesser extent feedback inhibition to MSNs in HD can potentially be sources for the increased GABA synaptic activity of indirect pathway MSNs.
Related JoVE Video
Related JoVE Video
The threat of disease increases as species move toward extinction.
Conserv. Biol.
PUBLISHED: 03-19-2013
Show Abstract
Hide Abstract
At local scales, infectious disease is a common driver of population declines, but globally it is an infrequent contributor to species extinction and endangerment. For species at risk of extinction from disease important questions remain unanswered, including when does disease become a threat to species and does it co-occur, predictably, with other threats? Using newly compiled data from the International Union for Conservation of Nature (IUCN) Red List, we examined the relative role and co-occurrence of threats associated with amphibians, birds, and mammals at 6 levels of extinction risk (i.e., Red List status categories: least concern, near threatened, vulnerable, endangered, critically endangered, and extinct in the wild/extinct). We tested the null hypothesis that the proportion of species threatened by disease is the same in all 6 Red List status categories. Our approach revealed a new method for determining when disease most frequently threatens species at risk of extinction. The proportion of species threatened by disease varied significantly between IUCN status categories and linearly increased for amphibians, birds, and all species combined as these taxa move from move from least concern to critically endangered. Disease was infrequently the single contributing threat. However, when a species was negatively affected by a major threat other than disease (e.g., invasive species, land-use change) that species was more likely to be simultaneously threatened by disease than species that had no other threats. Potential drivers of these trends include ecological factors, clustering of phylogenetically related species in Red List status categories, discovery bias among species at greater risk of extinction, and availability of data. We echo earlier calls for baseline data on the presence of parasites and pathogens in species when they show the first signs of extinction risk and arguably before. La Amenaza de Enfermedades Incrementa a Medida que las Especies se Aproximan a la Extinción.
Related JoVE Video
Effects of the Pimelic Diphenylamide Histone Deacetylase Inhibitor HDACi 4b on the R6/2 and N171-82Q Mouse Models of Huntingtons Disease.
PLoS Curr
PUBLISHED: 02-26-2013
Show Abstract
Hide Abstract
This report represents a detailed description of experiments designed to replicate and extend the findings of a published study on the effects of treating the R6/2 Huntingtons disease (HD) mouse model with ~300 CAG repeats using the pimelic diphenylamide histone deacetylase (HDAC) inhibitor, HDACi 4b (Thomas et al., 2008). In addition to testing the R6/2 mice, similar experiments examined the effects of the drug on a second transgenic HD mouse model, the N171-82Q mice. As in the original study, the drug was delivered in the drinking water. In the present study we tested larger groups of mice than in the original study. The results indicated that we were unable to replicate the significant behavioral effects of oral HDACi 4b treatment in the R6/2 mice. There were however, non-significant trends for the treated R6/2 mice to be less affected on some of the measures and there were instances of phenotype progression being delayed in these treated mice. In contrast, we did replicate the protection from striatal atrophy in the R6/2 mice. We also did not observe any beneficial effects of HDACi 4b treatment in the N171-82Q mice. Although the behavioral procedures were replicated and an automated activity assessment was added, there were several unexpected complications in terms of solubility of the drug, CAG repeat length differences and gender differences in progression of the phenotype that could have affected outcomes. Clearly more studies will have to be performed using other methods of delivery as well as assessing effects in more slowly progressing HD models to better evaluate the effects of this HDAC inhibitor.
Related JoVE Video
Quantitation of fixative-induced morphologic and antigenic variation in mouse and human breast cancers.
Lab. Invest.
PUBLISHED: 02-11-2013
Show Abstract
Hide Abstract
Quantitative Image Analysis (QIA) of digitized whole slide images for morphometric parameters and immunohistochemistry of breast cancer antigens was used to evaluate the technical reproducibility, biological variability, and intratumoral heterogeneity in three transplantable mouse mammary tumor models of human breast cancer. The relative preservation of structure and immunogenicity of the three mouse models and three human breast cancers was also compared when fixed with representatives of four distinct classes of fixatives. The three mouse mammary tumor cell models were an ER+/PR+ model (SSM2), a Her2+ model (NDL), and a triple negative model (MET1). The four breast cancer antigens were ER, PR, Her2, and Ki67. The fixatives included examples of (1) strong cross-linkers, (2) weak cross-linkers, (3) coagulants, and (4) combination fixatives. Each parameter was quantitatively analyzed using modified Aperio Technologies ImageScope algorithms. Careful pre-analytical adjustments to the algorithms were required to provide accurate results. The QIA permitted rigorous statistical analysis of results and grading by rank order. The analyses suggested excellent technical reproducibility and confirmed biological heterogeneity within each tumor. The strong cross-linker fixatives, such as formalin, consistently ranked higher than weak cross-linker, coagulant and combination fixatives in both the morphometric and immunohistochemical parameters.
Related JoVE Video
Maternal parenting styles and mother-child relationship among adolescents with and without persistent attention-deficit/hyperactivity disorder.
Res Dev Disabil
PUBLISHED: 02-01-2013
Show Abstract
Hide Abstract
We investigated mothering and mother-child interactions in adolescents with and without persistent attention-deficit/hyperactivity disorder (ADHD) in a sample of 190 adolescents with persistent DSM-IV ADHD, 147 without persistent ADHD, and 223 without ADHD. Both participants and their mothers received psychiatric interviews for diagnosis of ADHD and other mental disorders; and reported on the Parental Bonding Instrument about mothers parenting style, the Social Adjustment Inventory for Children and Adolescents for interactions with mothers and home behavioral problems. The mothers also reported on their ADHD and neurotic/depressive symptoms. Our results based on both informants showed that both ADHD groups obtained less affection/care and more overprotection and control from the mothers, and perceived less family support than those without ADHD. Childs inattention and comorbidity, and maternal depression were significantly correlated with decreased maternal affection/care and increased maternal controls; childs hyperactivity-impulsivity and maternal neurotic trait were significantly correlated with maternal overprotection; and childs inattention and comorbidity, and maternal neurotic/depressive symptoms were significantly correlated with impaired mother-child interactions and less family support. Our findings suggested that, regardless of persistence, childhood ADHD diagnosis, particularly inattention symptoms and comorbidity, combining with maternal neurotic/depressive symptoms was associated with impaired maternal process.
Related JoVE Video
The deficits on a cortical-subcortical loop of meaning processing in schizophrenia.
Neuroreport
PUBLISHED: 01-18-2013
Show Abstract
Hide Abstract
Thought disorder is a core symptom of schizophrenia. However, the underlying mechanism is not well understood. Functional MRI (fMRI) was used to examine the neural mechanism of thought disorder in 20 patients with schizophrenia and 20 healthy controls during semantic judgments. Two indexes of disorganized thought were further used to evaluate individual differences in thought disturbance in the patients. Compared with the controls, the patients showed greater activation in left inferior frontal gyrus (BA45) and reduced activation in the left caudate nucleus for meaning-related pairs. Moreover, in patients, effective connectivity from Dynamic Causal Modeling showed that the modulatory effect from the caudate nucleus to the inferior frontal gyrus was weaker than that in controls, indicating a disrupted cortical-subcortical language loop for semantic processing in patients. Finally, increasing scores of disorganized thought were correlated with greater activation in the inferior frontal gyrus and weaker connection strength from the caudate nucleus to the inferior frontal gyrus. Patients with more severe disorganized symptoms might receive less efficient regulation from the caudate nucleus, resulting in increased demands for the inferior frontal gyrus to retrieve or select semantic knowledge in the cortical-subcortical circuit.
Related JoVE Video
Dopamine imbalance in Huntingtons disease: a mechanism for the lack of behavioral flexibility.
Front Neurosci
PUBLISHED: 01-01-2013
Show Abstract
Hide Abstract
Dopamine (DA) plays an essential role in the control of coordinated movements. Alterations in DA balance in the striatum lead to pathological conditions such as Parkinsons and Huntingtons diseases (HD). HD is a progressive, invariably fatal neurodegenerative disease caused by a genetic mutation producing an expansion of glutamine repeats and is characterized by abnormal dance-like movements (chorea). The principal pathology is the loss of striatal and cortical projection neurons. Changes in brain DA content and receptor number contribute to abnormal movements and cognitive deficits in HD. In particular, during the early hyperkinetic stage of HD, DA levels are increased whereas expression of DA receptors is reduced. In contrast, in the late akinetic stage, DA levels are significantly decreased and resemble those of a Parkinsonian state. Time-dependent changes in DA transmission parallel biphasic changes in glutamate synaptic transmission and may enhance alterations in glutamate receptor-mediated synaptic activity. In this review, we focus on neuronal electrophysiological mechanisms that may lead to some of the motor and cognitive symptoms of HD and how they relate to dysfunction in DA neurotransmission. Based on clinical and experimental findings, we propose that some of the behavioral alterations in HD, including reduced behavioral flexibility, may be caused by altered DA modulatory function. Thus, restoring DA balance alone or in conjunction with glutamate receptor antagonists could be a viable therapeutic approach.
Related JoVE Video
Noninvasive electroanatomic mapping of human ventricular arrhythmias with electrocardiographic imaging.
Sci Transl Med
PUBLISHED: 09-03-2011
Show Abstract
Hide Abstract
The rapid heartbeat of ventricular tachycardia (VT) can lead to sudden cardiac death and is a major health issue worldwide. Efforts to identify patients at risk, determine mechanisms of VT, and effectively prevent and treat VT through a mechanism-based approach would all be facilitated by continuous, noninvasive imaging of the arrhythmia over the entire heart. Here, we present noninvasive real-time images of human ventricular arrhythmias using electrocardiographic imaging (ECGI). Our results reveal diverse activation patterns, mechanisms, and sites of initiation of human VT. The spatial resolution of ECGI is superior to that of the routinely used 12-lead electrocardiogram, which provides only global information, and ECGI has distinct advantages over the currently used method of mapping with invasive catheter-applied electrodes. The spatial resolution of this method and its ability to image electrical activation sequences over the entire ventricular surfaces in a single heartbeat allowed us to determine VT initiation sites and continuation pathways, as well as VT relationships to ventricular substrates, including anatomical scars and abnormal electrophysiological substrate. Thus, ECGI can map the VT activation sequence and identify the location and depth of VT origin in individual patients, allowing personalized treatment of patients with ventricular arrhythmias.
Related JoVE Video
The effects of acellular amniotic membrane matrix on osteogenic differentiation and ERK1/2 signaling in human dental apical papilla cells.
Biomaterials
PUBLISHED: 08-31-2011
Show Abstract
Hide Abstract
The amniotic membrane (AM) has been widely used in the field of tissue engineering because of the favorable biological properties for scaffolding material. However, little is known about the effects of an acellular AM matrix on the osteogenic differentiation of mesenchymal stem cells. In this study, it was found that both basement membrane side and collagenous stroma side of the acellular AM matrix were capable of providing a preferential environment for driving the osteogenic differentiation of human dental apical papilla cells (APCs) with proven stem cell characteristics. Acellular AM matrix potentiated the induction effect of osteogenic supplements (OS) such as ascorbic acid, ?-glycerophosphate, and dexamethasone and enhanced the osteogenic differentiation of APCs, as seen by increased core-binding factor alpha 1 (Cbfa-1) phosphorylation, alkaline phosphatase activity, mRNA expression of osteogenic marker genes, and mineralized matrix deposition. Even in the absence of soluble OS, acellular AM matrix also could exert the substrate-induced effect on initiating APCs differentiation. Especially, the collagenous stroma side was more effective than the basement membrane side. Moreover, the AM-induced effect was significantly inhibited by U0126, an inhibitor of extracellular signaling-regulated kinase 1/2 (ERK1/2) signaling. Taken together, the osteogenic differentiation promoting effect on APCs is AM-specific, which provides potential applications of acellular AM matrix in bone/tooth tissue engineering.
Related JoVE Video
Intracellular trafficking of polyamidoamine-poly(ethylene glycol) block copolymers in DNA delivery.
Bioconjug. Chem.
PUBLISHED: 07-15-2011
Show Abstract
Hide Abstract
The delivery of nucleic acids has the potential to revolutionize medicine by allowing previously untreatable diseases to be clinically addressed. Viral delivery systems have shown immunogenicity and toxicity dangers, but synthetic vectors have lagged in transfection efficiency. Previously, we developed a modular, linear-dendritic block copolymer architecture with high gene transfection efficiency compared to commercial standards. This rationally designed system makes use of a cationic dendritic block to condense the anionic DNA and forms complexes with favorable endosomal escape properties. The linear block provides biocompatibility and protection from serum proteins, and can be functionalized with a targeting ligand. In this work, we quantitate performance of this system with respect to intracellular barriers to gene delivery using both high-throughput and traditional approaches. An image-based, high-throughput assay for endosomal escape is described and applied to the block copolymer system. Nuclear entry is demonstrated to be the most significant barrier to more efficient delivery and will be addressed in future versions of the system.
Related JoVE Video
Does the hikikomori syndrome of social withdrawal exist outside Japan? A preliminary international investigation.
Soc Psychiatry Psychiatr Epidemiol
PUBLISHED: 06-06-2011
Show Abstract
Hide Abstract
To explore whether the hikikomori syndrome (social withdrawal) described in Japan exists in other countries, and if so, how patients with the syndrome are diagnosed and treated.
Related JoVE Video
Landmark identification errors on cone-beam computed tomography-derived cephalograms and conventional digital cephalograms.
Am J Orthod Dentofacial Orthop
PUBLISHED: 06-01-2011
Show Abstract
Hide Abstract
In this study, we investigated the landmark identification errors on cone-beam computed tomography (CBCT)-derived cephalograms and conventional digital cephalograms.
Related JoVE Video
Enhanced GABAergic network and receptor function in pediatric cortical dysplasia Type IIB compared with Tuberous Sclerosis Complex.
Neurobiol. Dis.
PUBLISHED: 05-13-2011
Show Abstract
Hide Abstract
Tuberous Sclerosis Complex (TSC) and cortical dysplasia Type IIB (CDIIB) share histopathologic features that suggest similar epileptogenic mechanisms. This study compared the morphological and electrophysiological properties of cortical cells in tissue from pediatric TSC (n=20) and CDIIB (n=20) patients using whole-cell patch clamp recordings and biocytin staining. Cell types were normal-appearing and dysmorphic-cytomegalic pyramidal neurons, interneurons, and giant/balloon cells, including intermediate neuronal-glial cells. In the cortical mantle, giant/balloon cells occurred more frequently in TSC than in CDIIB cases, whereas cytomegalic pyramidal neurons were found more frequently in CDIIB. Cell morphology and membrane properties were similar in TSC and CDIIB cases. Except for giant/balloon and intermediate cells, all neuronal cell types fired action potentials and displayed spontaneous postsynaptic currents. However, the frequency of spontaneous glutamatergic postsynaptic currents in normal pyramidal neurons and interneurons was significantly lower in CDIIB compared with TSC cases and the GABAergic activity was higher in all neuronal cell types in CDIIB. Further, acutely dissociated pyramidal neurons displayed higher sensitivity to exogenous application of GABA in CDIIB compared with TSC cases. These results indicate that, in spite of similar histopathologic features and basic cell membrane properties, TSC and CDIIB display differences in the topography of abnormal cells, excitatory and inhibitory synaptic network properties, and GABA(A) receptor sensitivity. These differences support the notion that the mechanisms of epileptogenesis could differ in patients with TSC and CDIIB. Consequently, pharmacologic therapies should take these findings into consideration.
Related JoVE Video
Air kerma standard and measurement comparison on source strength determination for high-dose rate 192Ir brachytherapy in Taiwan.
Radiat Prot Dosimetry
PUBLISHED: 04-16-2011
Show Abstract
Hide Abstract
This paper describes the establishment by the Institute of Nuclear Energy Research (INER, Taiwan) of the reference air kerma rate (RAKR) calibration standard for measurement of high-dose rate (HDR) (192)Ir brachytherapy source strength. A bilateral comparison has been made in the RAKR standards for HDR (192)Ir brachytherapy sources at the INER and Physikalisch-Technische Bundesanstalt (PTB, Germany) and the measurement difference was within the overall standard uncertainty and showed good agreement between the two calibration standard systems established at the INER and the PTB. Besides, INER also worked with 20 domestic hospitals to organise an on-site measurement comparison programme to explore the status of HDR (192)Ir brachytherapy source strength determination in Taiwan. The comparison results presented the ratios of RAKR with vendor values, as determined by INER and hospitals from the programme. The ratios fall in all cases within the ± 3 % guaranteed by the vendors for a coverage factor of k = 2 or at 95 % confidence level.
Related JoVE Video
Introducing the concept of modern depression in Japan; an international case vignette survey.
J Affect Disord
PUBLISHED: 03-28-2011
Show Abstract
Hide Abstract
Japanese psychiatrists have increasingly reported patients with depression that does not seem to fit the criteria of the ICD-10 and the DSM-IV, and which has recently been called modern type depression (MTD). We explored whether MTD is frequently seen in Japan and also in other countries, and if so, how patients with MTD are diagnosed and treated.
Related JoVE Video
High-sensitivity electrochemical enzyme-linked assay on a microfluidic interdigitated microelectrode.
Biosens Bioelectron
PUBLISHED: 03-03-2011
Show Abstract
Hide Abstract
A novel enzyme-linked DNA hybridization assay on an interdigitated array (IDA) microelectrode integrated into a microfluidic channel is demonstrated with sub-nM detection limit. To improve the detection limit as compared to conventional electrochemical biosensors, a recyclable redox product, 4-aminophenol (PAP) is used with an IDA microelectrode. The IDA has a modest and easily fabricated inter-digit spacing of 10 ?m, yet we were able to demonstrate 97% recycling efficiency of PAP due to the integration in a microfluidic channel. With a 70 nL sample volume, the characterized detection limit for PAP of 1.0 × 10?¹? M is achieved, with a linear dynamic range that extends from 1.0 × 10?? to 1.0 × 10?? M. This detection limit, which is the lowest reported detection limit for PAP, is due to the increased sensitivity provided by the sample confinement in the microfluidic channel, as well as the increased repeatability due to perfectly static flow in the microchannel and an additional anti-fouling step in the protocol. DNA sequence detection is achieved through a hybridization sandwich of an immobilized complementary probe, the target DNA sequence, and a second complementary probe labeled with ?-galactosidase (?-GAL); the ?-GAL converts its substrate, 4-aminophenyl-d-galactopyranoside (PAPG), into PAP. In this report we present the lowest reported observed detection limit (1.0 × 10?¹? M) for an enzyme-linked DNA hybridization assay using an IDA microelectrode and a redox signaling paradigm. Thus, we have demonstrated highly sensitive detection of a targeted DNA sequence using a low-cost easily fabricated electrochemical biosensor integrated into a microfluidic channel.
Related JoVE Video
Epigenetic effects and molecular mechanisms of tumorigenesis induced by cigarette smoke: an overview.
J Oncol
PUBLISHED: 01-24-2011
Show Abstract
Hide Abstract
Cigarette smoking is one of the major causes of carcinogenesis. Direct genotoxicity induced by cigarette smoke leads to initiation of carcinogenesis. Nongenotoxic (epigenetic) effects of cigarette smoke also act as modulators altering cellular functions. These two effects underlie the mechanisms of tumor promotion and progression. While there is no lack of general reviews on the genotoxic and carcinogenic potentials of cigarette smoke in lung carcinogenesis, updated review on the epigenetic effects and molecular mechanisms of cigarette smoke and carcinogenesis, not limited to lung, is lacking. We are presenting a comprehensive review of recent investigations on cigarette smoke, with special attentions to nicotine, NNK, and PAHs. The current understanding on their molecular mechanisms include (1) receptors, (2) cell cycle regulators, (3) signaling pathways, (4) apoptosis mediators, (5) angiogenic factors, and (6) invasive and metastasis mediators. This review highlighted the complexity biological responses to cigarette smoke components and their involvements in tumorigenesis.
Related JoVE Video
Longitudinal investigation of permeability and distribution of macromolecules in mouse malignant transformation using PET.
Clin. Cancer Res.
PUBLISHED: 11-24-2010
Show Abstract
Hide Abstract
We apply positron emission tomography (PET) to elucidate changes in nanocarrier extravasation during the transition from premalignant to malignant cancer, providing insight into the use of imaging to characterize early cancerous lesions and the utility of nanoparticles in early disease.
Related JoVE Video
Assessment of carprofen and buprenorphine on recovery of mice after surgical removal of the mammary fat pad.
J. Am. Assoc. Lab. Anim. Sci.
PUBLISHED: 09-23-2010
Show Abstract
Hide Abstract
The purpose of this study was to determine the level of pain elicited by mammary fat pad removal surgery and the effects of postoperative analgesics on recovery. Female FVB mice were anesthetized, and mammary fat pad removal was performed. After surgery, mice received carprofen, buprenorphine, a combination of carprofen and buprenorphine, or saline treatment. Additional mice received anesthesia but no surgery or treatment. Food and water intake, body weight, wheel running activity, and a visual assessment score were recorded daily for 4 d after surgery and compared with presurgical findings. Corticosterone metabolites in fecal samples were analyzed at 12 and 24 h postsurgically and compared with baseline values. All surgical groups had significantly decreased food intake at 24 h, with a return to baseline by 48 h. The combination treatment resulted in a significantly decreased water intake and body weight at 24 h. All surgical groups had significantly decreased wheel running activity at 24 h only. The visual assessment scores indicated mild pain for all surgical groups, with the buprenorphine treated mice showing the highest pain index scores, as compared with nonsurgical controls. Fecal corticosterone metabolite levels did not differ significantly between any of the groups or across time. The parameters used in this study did not indicate that administration of these analgesic regimens improved recovery as compared with that of saline-treated mice. Care should be taken when using visual assessment scores to evaluate pain in mice, given that analgesics may have side effects that inadvertently elevate the score.
Related JoVE Video
Rescuing the Corticostriatal Synaptic Disconnection in the R6/2 Mouse Model of Huntingtons Disease: Exercise, Adenosine Receptors and Ampakines.
PLoS Curr
PUBLISHED: 09-22-2010
Show Abstract
Hide Abstract
In the R6/2 mouse model of Huntingtons disease (HD) we examined the effects of a number of behavioral and pharmacological manipulations aimed at rescuing the progressive loss of synaptic communication between cerebral cortex and striatum. Two cohorts of transgenic mice with ~110 and 210 CAG repeats were utilized. Exercise prevented the reduction in striatal medium-sized spiny neuron membrane capacitance but did not reestablish synaptic communication. Activation of adenosine A2A type receptors renormalized postsynaptic activity to some extent. Finally, the ampakine Cx614, which has been shown to prevent ?-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA) receptor desensitization, slow deactivation, and facilitate glutamate release, induced significant increases in synaptic activity, albeit the effect was somewhat reduced in fully symptomatic, compared to control mice. With some limitations, each of these strategies can be used to delay and partially rescue phenotypic progression of HD in this model.
Related JoVE Video
Development of a method for the determination of bisphenol A at trace concentrations in human blood and urine and elucidation of factors influencing method accuracy and sensitivity.
J Anal Toxicol
PUBLISHED: 07-29-2010
Show Abstract
Hide Abstract
Bisphenol A (BPA) is an industrial chemical used to make polymers including some used in food contact applications. Virtually complete presystemic clearance of orally administered BPA occurs in humans by metabolism to BPA-glucuronide (BPA-G), but some biomonitoring studies report low concentrations of free (parent) BPA in human blood and urine. Trace contamination of BPA from exogenous sources or hydrolysis of BPA-G to free BPA, either during or after biomonitoring specimen collection, may have contributed to the reported concentrations of free BPA. An analytical method for the determination of free BPA in human blood and urine was developed and validated in two independent laboratories, using the latest generation of high-performance liquid chromatography-tandem mass spectrometry instrumentation to ensure the desired high sensitivity and selectivity. The method was designed to account for and/or eliminate background contamination from all sources and demonstrated that contamination could occur from devices used for specimen collection or storage, as well as other sources. The method employed an internal standard (BPA-d(8)) and demonstrated accuracy and reproducibility in both matrices fortified with BPA or a surrogate analyte ((13)C-BPA) at a low quantitation limit (0.1-0.2 ng/mL). For validation, five replicate samples were analyzed to evaluate reproducibility. Importantly, it was demonstrated that the conditions of the method did not result in the hydrolysis of BPA-G to free BPA, another possible source of error in BPA analysis. Application of the principles defined by this method will be critical to assure valid analytical results in any future biomonitoring studies.
Related JoVE Video
Potentiation of Th17 cytokines in aging process contributes to the development of colitis.
Cell. Immunol.
PUBLISHED: 07-15-2010
Show Abstract
Hide Abstract
Th17 cells, which produce IL-17 and IL-22, promote autoimmunity in mice and have been implicated in the pathogenesis of autoimmune/inflammatory diseases in humans. However, the Th17 immune response in the aging process is still not clear. In the present study, we found that the induction of IL-17-producing CD4(+) T cells was significantly increased in aged individuals compared with young healthy ones. The mRNA expression of IL-17, IL-17F, IL-22, and RORC2 was also significantly increased in aged people. Similar to humans, Th17 cells as well as mRNAs encoding IL-17, IL-22 and ROR?t were dramatically elevated in naïve T cells from aged mouse compared to young ones. In addition, CD44 positive IL-17-producing CD4(+) T cells were significantly higher in aged mice, suggesting that memory T cells are an important source of IL-17 production. Furthermore, the percentage of IL-17-producing CD4(+) T cells generated in co-culture with dendritic cells from either aged or young mice did not show significant differences, suggesting that dendritic cells do not play a primary role in the elevation of Th17 cytokines in aged mouse cells. Importantly, transfer of CD4(+)CD45Rb(hi) cells from aged mice induced more severe colitis in RAG(-/-) mice compared to cells from young mice, Taken together, these results suggest that Th17 immune responses are elevated in aging humans and mice and may contribute to the increased development of inflammatory disorders in the elderly.
Related JoVE Video
Comparative study of cellular and synaptic abnormalities in brain tissue samples from pediatric tuberous sclerosis complex and cortical dysplasia type II.
Epilepsia
PUBLISHED: 07-13-2010
Show Abstract
Hide Abstract
Tuberous sclerosis complex (TSC) and severe cortical dysplasia (CD), or CD type II according to Palmini classification, share histopathologic similarities, specifically the presence of cytomegalic neurons and balloon cells. In this study we examined the morphologic and electrophysiologic properties of cells in cortical tissue samples from pediatric patients with TSC and CD type II who underwent surgery for pharmacoresistant epilepsy. Normal-appearing pyramidal neurons from TSC and CD type II cases had similar passive membrane properties. However, the frequency of excitatory postsynaptic currents (EPSCs) was higher in neurons from TSC compared to severe CD cases, particularly the frequency of medium- and large-amplitude synaptic events. In addition, EPSCs rise and decay times were slower in normal cells from TSC compared to severe CD cases. Balloon cells were found more frequently in TSC cases, whereas cytomegalic pyramidal neurons occurred more often in CD type II cases. Both cell types were similar morphologically and electrophysiologically in TSC and severe CD. These results suggest that even though the histopathology in TSC and severe CD is similar, there are subtle differences in spontaneous synaptic activity and topographic distribution of abnormal cells. These differences may contribute to variable mechanisms of epileptogenesis in patients with TSC compared with CD type II.
Related JoVE Video
Pterostilbene induces autophagy and apoptosis in sensitive and chemoresistant human bladder cancer cells.
Mol Nutr Food Res
PUBLISHED: 07-07-2010
Show Abstract
Hide Abstract
Bladder cancer is one of the most common malignancies in the world. The majority of bladder cancer deaths are due to unresectable lesions that are resistant to chemotherapy. Pterostilbene (PT), a naturally occurring phytoalexin, possesses a variety of pharmacologic activities, including antioxidant, cancer prevention activity and cytotoxicity to many cancers. We found that PT effectively inhibits the growth of sensitive and chemoresistant human bladder cancer cells by inducing cell cycle arrest, autophagy and apoptosis. Down-regulations of Cyclin A, B and D1 and pRB are the results of PT-induced cell cycle arrest.
Related JoVE Video
Glucosamine promotes osteogenic differentiation of dental pulp stem cells through modulating the level of the transforming growth factor-beta type I receptor.
J. Cell. Physiol.
PUBLISHED: 05-12-2010
Show Abstract
Hide Abstract
Dental pulp stem cells (DPSCs) are clonogenic, self-renewing, and multi-potential DPSCs capable of differentiating into osteoblasts. In this study, primary cell cultures were obtained from human dental pulp tissue of developing third molars, and flow cytometry was used to sort the subpopulation of DPSCs with STRO-1 and CD146 double-positive expression (denoted "DPSCs"). It was noted that DPSCs exhibited superior clonogenic potential and osteogenic differentiation capability than the dental pulp cell subpopulation with STRO-1 and CD146 double-negative expression (denoted DPCs). Furthermore, a low concentration (0.005 mg/ml) of exogenous glucosamine (GlcN) was effective in promoting the early osteogenic differentiation of DPSCs through the transforming growth factor-beta receptor (TGF-betar) type I and Smads signal pathways, which upregulated the Runt-related transcription factor 2/core-binding factor alpha1 (Runx2/Cbfa1) and alkaline phosphatase at both the mRNA and protein levels. In the presence of osteogenic supplements, GlcN-treated DPSCs produced more mineralized-matrix deposition than did the untreated groups. Taken together, this study demonstrates the capacity of GlcN to promote the osteogenic differentiation of human DPSCs, and the underlying mechanism involves a TGF-betar-dependent Smad signal pathway.
Related JoVE Video
Long-term nicotine exposure-induced chemoresistance is mediated by activation of Stat3 and downregulation of ERK1/2 via nAChR and beta-adrenoceptors in human bladder cancer cells.
Toxicol. Sci.
PUBLISHED: 01-27-2010
Show Abstract
Hide Abstract
Previous reports suggested that bladder cancer patients who continue to smoke while receiving chemotherapy have poorer outcomes than their nonsmoking counterparts. Nicotine, the major addictive compound in cigarette smoke, is known to induce chemoresistance in some cancer cells. Chemoresistance has been linked to the activation of Stat3 (signal transducer and activator of transcription). The objective of this study was to identify the role of Stat3 in chemoresistance induced by nicotine in human bladder cancer cell line, T24 cells. Chemoresistant T24 cells were established by persistent nicotine treatment. Apoptosis and cell cycle parameters were analyzed by Annexin V staining, poly(ADP-ribose) polymerase degradation, caspase activity, and propidium iodide staining. Signal transduction mediating the chemoresistance was detected by Western blotting and small interfering RNA (siRNA) transfection. We provide evidence for the first time that nicotine strongly activated Stat3, leading to Cyclin D1 overexpression, cell cycle perturbations, and chemoresistance. Furthermore, nicotine mobilized Stat3 signaling, resulting in the loss of extracellular signal-regulated protein kinase 1/2 (ERK 1/2) activation and reduced chemosensitivity via nicotinic acetylcholine receptors and beta-adrenoceptors. Inhibition of Stat3 by siRNA or a specific inhibitor restored chemosensitivity in T24 cells. Stat3 could be the major target for increasing chemosensitivity in patients who develop chemoresistance during chemotherapy, and avoidance of cigarette smoking or nicotine-based treatments may increase the efficacy of chemotherapy.
Related JoVE Video
Hepatic heme-regulated inhibitor (HRI) eukaryotic initiation factor 2alpha kinase: a protagonist of heme-mediated translational control of CYP2B enzymes and a modulator of basal endoplasmic reticulum stress tone.
Mol. Pharmacol.
PUBLISHED: 01-13-2010
Show Abstract
Hide Abstract
We have reported previously that the hepatic heme-regulated inhibitor (HRI)-eukaryotic initiation factor 2 alpha (eIF2 alpha) kinase is activated in acute heme-deficient states, resulting in translational shut-off of global hepatic protein synthesis, including phenobarbital (PB)-mediated induction of CYP2B enzymes in rats. These findings revealed that heme regulates hepatic CYP2B synthesis at the translational level via HRI. As a proof of concept, we have now employed a genetic HRI-knockout (KO) mouse hepatocyte model. In HRI-KO hepatocytes, PB-mediated CYP2B protein induction is no longer regulated by hepatic heme availability and proceeds undeterred even after acute hepatic heme depletion. It is noteworthy that genetic ablation of HRI led to a small albeit significant elevation of basal hepatic endoplasmic reticulum (ER) stress as revealed by the activation of ER stress-inducible RNA-dependent protein kinase-like ER-integral (PERK) eIF2 alpha-kinase, and induction of hepatic protein ubiquitination and ER chaperones Grp78 and Grp94. Such ER stress was further augmented after PB-mediated hepatic protein induction. These findings suggest that HRI normally modulates the basal hepatic ER stress tone. Furthermore, because HRI exists in both human and rat liver in its heme-sensitive form and is inducible by cytochrome P450 inducers such as PB, these findings are clinically relevant to acute heme-deficient states, such as the acute hepatic porphyrias. Activation of this exquisitely sensitive heme sensor would normally protect cells by safeguarding cellular energy and nutrients during acute heme deficiency. However, similar HRI activation in genetically predisposed persons could lead to global translational arrest of physiologically relevant enzymes and proteins, resulting in the severe and often fatal clinical symptoms of the acute hepatic porphyrias.
Related JoVE Video
Proficiency testing feasibility study for the measurement of gamma-emitting clearance samples.
Appl Radiat Isot
PUBLISHED: 01-06-2010
Show Abstract
Hide Abstract
A proficiency testing feasibility study program was proposed by the National Radiation Standard Laboratory (NRSL) of Taiwan to understand the capabilities of laboratories dealing with clearance measurements, and to issue related technical criteria for radioactive waste assay. In this program, twelve blind test samples with different levels of radioactivity, radionuclides and different packing densities were prepared. Seven laboratories participated in this program and fourteen instruments were tested. Participants were required to report their raw data to NRSL, which would evaluate the effects of the background, geometry and the packing density to obtain the final results of the participants. In this study, the typical uncertainties of the participants were around 24%, and about 70% of the measured results produced E(n) values, which were smaller than one.
Related JoVE Video
Lab-on-chip flow injection analysis system without an external pump and valves and integrated with an in line electrochemical detector.
Anal. Chem.
PUBLISHED: 11-21-2009
Show Abstract
Hide Abstract
Surface energy in small droplets can be used to drive samples through microchannels. When a sample fluid is spontaneously driven through a solution filled microchannel with liquid droplets on its entry (sample) and exit (reservoir) ports, it is termed as a passive pumping device. A passive pump driven microfluidic system integrated with microfabricated planar electrodes or electrode arrays (e.g., interdigitated electrode arrays or microband electrode arrays) can be considered as a manifold for flow injection analysis without an external pump and injector valve. Factors affecting the passive pump driven flow rate in a polydimethylsiloxane (PDMS)-glass hybrid microfluidic system, including the volume, viscosity, and surface tension of the sample solution and the tilt of the microfluidic channel, are analyzed. By placing 2 microL of hexacyanoferrate (II) solutions at the entry port of the device, peak shaped transients were recorded. The peak heights showed linear dependence on the sample concentrations between 3 x 10(-7) and 10(-5) M.
Related JoVE Video
Formation of salivary acinar cell spheroids in vitro above a polyvinyl alcohol-coated surface.
J Biomed Mater Res A
PUBLISHED: 11-05-2009
Show Abstract
Hide Abstract
Tissue engineering of salivary glands offers the potential for future use in the treatment of patients with salivary hypofunction. Biocompatible materials that promote acinar cell aggregation and function in vitro are an essential part of salivary gland tissue engineering. In this study, rat parotid acinar cells assembled into three-dimensional aggregates above the polyvinyl alcohol (PVA)-coated surface. These aggregates developed compact acinar cell spheroids resembling in vivo physiological condition, which were different from the traditional monolayered morphology in vitro. Cells remained viable and with better functional activity in response to acetylcholine in the spheroids and could form monolayered acinar cells when they were reinoculated on tissue culture polystyrene wells. To interpret the phenomenon further, we proposed that the formation of acinar cell spheroids on the PVA is mediated by a balance between two competing forces: the interactions of cell-PVA and cell-cell. This study demonstrated the formation of functional cell spheroids above a PVA-coated surface may provide an in vitro system for investigating cell behaviors for tissue engineering of artificial salivary gland.
Related JoVE Video
Interactive effects of mechanical stretching and extracellular matrix proteins on initiating osteogenic differentiation of human mesenchymal stem cells.
J. Cell. Biochem.
PUBLISHED: 10-02-2009
Show Abstract
Hide Abstract
Human mesenchymal stem cells (hMSCs) are characterized by their abilities to differentiate into different lineages, including osteoblasts. Besides soluble factors, mechanical strain and extracellular matrix (ECM) proteins play important roles in osteogenic differentiation of hMSCs. However, interactions between them are still not fully understood. The purpose of this study was to investigate the combined effects of insoluble chemical and mechanical factors (ECM proteins vs. cyclic stretching) in driving hMSCs into osteogenic differentiation. To avoid the influence from osteogenic supplements, hMSCs were cultured in regular medium and subjected to cyclic mechanical stretching using a Flexcell Tension system (3% elongation at 0.1 Hz) when they were grown on substrates coated with various ECM proteins (collagen I (Col I), vitronectin (VN), fibronectin (FN), and laminin (LN)). Using alkaline phosphatase (ALP) activity and mineralized matrix deposition as respective indicators of the early and late stages of osteogenesis, we report herein that all of the ECM proteins tested supported hMSC differentiation into osteogenic phenotypes in the absence of osteogenic supplements. Moreover, cyclic mechanical stretching activated the phosphorylation of focal adhesion kinase (FAK), upregulated the transcription and phosphorylation of core-binding factor alpha-1 (Cbfa1), and subsequently increased ALP activity and mineralized matrix deposition. Among the ECM proteins tested, FN and LN exhibited greater effects of supporting stretching-induced osteogenic differentiation than did Col I and VN. The ability of ECM proteins and mechanical stretching to regulate osteogenesis in hMSCs can be exploited in bone tissue engineering via approximate matrix design or application of mechanical stimulation.
Related JoVE Video
Development of improved free-air ionisation chamber as absolute dosimetry standard for low-energy X rays in INER.
Radiat Prot Dosimetry
PUBLISHED: 09-29-2009
Show Abstract
Hide Abstract
The National Radiation Standard Laboratory of the Institute of Nuclear Energy Research (INER) designed and constructed an improved Attix style free-air ionisation chamber (FAC) for low-energy X-ray measurements. Clinically, X rays in this energy range are used in mammography radiology. This chamber is also used to perform air-kerma measurements. The original Attix two-sectional design was redesigned by INER using the piston design. The correction factors were determined experimentally for volume estimation, ion recombination and air attenuation. The aperture transmission, wall transmission, electron loss and photon scatter correction factors were determined using Monte Carlo calculations. INER established the Bureau International des Poids et Mesures (BIPM) X-ray beam code and performed a comparison of secondary standard air-kerma calibration factors for 10-50 kV low- energy X rays to verify the experimental accuracy and measurement consistency of the improved chamber. The INER-NMIJ/National Institute of Advanced Industrial Science and Technology (AIST) experimental results comparison using a transfer chamber yielded a difference <1.0% at the 95% confidence level in calibration factors. The overall uncertainty for the X-ray measurement in terms of air kerma was <0.6% at the 95% confidence level. These results indicated that the improved FAC is capable of serving as a primary standard as well as a trace standard in low-energy X-ray calibration services in Taiwan and even forming a basis for the future mammography X-ray air-kerma primary standard.
Related JoVE Video
Emotional management and 5-HT2A receptor gene variance in patients with schizophrenia.
Biol Psychol
PUBLISHED: 07-25-2009
Show Abstract
Hide Abstract
Individuals with schizophrenia exhibit impaired social cognitive functions, particularly emotion management. Emotion management may be partially regulated by the serotoninergic system; the -1438 A/G polymorphism in the promoter region of the 5-HT2A gene can modulate 5-HT2A activity and is linked to certain emotional traits and anger- and aggression-related behaviors. The current study aimed to investigate whether this 5-HT2A genetic variance is associated with social cognitive function, particularly the management of emotions. One hundred and fifteen patients with chronic schizophrenia were stabilized with an optimal-dose of antipsychotic treatment. All were genotyped for the -1438 A/G polymorphism and assessed with symptom rating scales, neurocognitive instruments, and the "Managing Emotions" section of Mayer-Salovey-Caruso Emotional Intelligence Test (MSCEIT). Multiple regression showed that patients with the A/G genotype performed better than those with G/G in managing emotion (p=0.018) but did not differ from those with the A/A genotype. Regarding the two subtasks of the Managing Emotions section, the A/G heterozygotes also performed better than the G/G homozygotes in the emotion management (p=0.026) and emotional relations (p=0.027) subtasks. The results suggest that variability in the 5-HT2A gene may influence emotion management in patients with schizophrenia.
Related JoVE Video
Clinical benefits of remote versus transtelephonic monitoring of implanted pacemakers.
J. Am. Coll. Cardiol.
PUBLISHED: 06-14-2009
Show Abstract
Hide Abstract
The purpose of this study was to evaluate remote pacemaker interrogation for the earlier diagnosis of clinically actionable events compared with traditional transtelephonic monitoring and routine in-person evaluation.
Related JoVE Video
Omega-3 Polyunsaturated Fatty Acids (n-3 PUFAs) in Cardiovascular Diseases (CVDs) and Depression: The Missing Link?
Cardiovasc Psychiatry Neurol
PUBLISHED: 04-29-2009
Show Abstract
Hide Abstract
Background. Based on epidemiological data, clinical trials, and meta-analytic reviews, omega-3 polyunsaturated fatty acids (n-3 PUFAs) seem to be a biological link between depression and cardiovascular diseases (CVDs). Presentation. Involvement of n-3 PUFAs in depression and CVDs may be associated with a chronic, low-grade, inflammation. We hypothesize that n-3 PUFAs link depression and CVDs via "PUFA-prostaglandin E2 (PGE2) cascade." Testing. To further support our hypothesis, case-control studies are needed to test the role of COX2 and PLA2 functions in depression and in CVDs. In addition, the effects of n-3 PUFAs on cardiovascular markers in depression and on depressive symptoms in CVDs should be investigated in clinical trials. Finally, the effects of manipulating COX2 and PLA2 functions on depression-like behaviors and cardiovascular functions could be explored in animal studies. Implications. n-3 PUFAs might be a promising treatment for both cardiovascular diseases and depression via its anti-inflammatory, cardioprotective, and neuroprotective effects.
Related JoVE Video
Dosimetry study for beta-radiation treatment of in-stent restenosis.
Radiat Prot Dosimetry
PUBLISHED: 04-17-2009
Show Abstract
Hide Abstract
Intravascular brachytherapy (IVBT) has been recognised as a treatment modality for reducing coronary restenosis after angioplasty and stent-implantation procedures. For the treatment of in-stent restenosis using beta-emitter (188)Re, delivering adequate doses to the entire vessel wall is not possible without the potential of overdosing tissues. A method to measure the dose distribution, perturbation and percentage depth dose using plane-parallel and cylindrical tissue-equivalent phantoms has been developed. Good agreement was found between experimental results and Monte Carlo simulation performed using MCNP4C code. The dose given to the affected area in the vascular region for intravascular radiation treatment was 15-30 Gy. Dose inhomogeneity beyond the stent surface decreased significantly with increasing radial distance. In the region close to the stent outer surface (>0.5-mm radial distance), a dose reduction of 11-17% due to the stent was observed. However, the dose perturbations due to the physical properties of metallic stents were found to be significant in IVBT for in-stent restenosis by using measured dose profiles in phantoms. The method can provide accuracy in beta isotope in vivo dosimetry results for treatments involving short-range dose distributions and provide a relatively high-level spatial resolution for detection.
Related JoVE Video
Comparative cytotoxicity of five current dentin bonding agents: role of cell cycle deregulation.
Acta Biomater
PUBLISHED: 03-07-2009
Show Abstract
Hide Abstract
To compare the cytotoxicity of three nano-dentin bonding agents (nano-DBAs) and two non-nano-DBAs using Chinese hamster ovary (CHO-K1) cells. We found that nano fillers were not the major contributing factor in DBA cytotoxicity, as analyzed by colony forming assay and 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Exposure of CHO-K1 cells to all three tested total-etching DBAs led to G(0)/G(1) cell cycle arrest, whereas exposure to higher concentrations of two tested nano-DBAs induced G(2)/M arrest. All five DBAs further induced apoptosis at the highest concentration, as analyzed by propidium iodide staining flow cytometry. The toxicity of all DBAs (1:4000v/v or higher) is related to increased reactive oxygen species (ROS) production, as analyzed by single cell DCF fluorescence flow cytometry. These results indicate that clinical application of DBAs may be potentially toxic to dental pulp tissues. Cytotoxicity of DBAs is associated with ROS production, cell cycle deregulation and apoptosis. Presence of methacrylate monomers such as PENTA and UDMA is possibly the major cytotoxic factor for DBAs. Further studies on other toxicological endpoints of nano-DBAs are necessary to highlight their safe use.
Related JoVE Video
Clearance measurement of metal scraps for nuclear facility at INER in Taiwan.
Appl Radiat Isot
PUBLISHED: 01-30-2009
Show Abstract
Hide Abstract
The nuclear facilities at Institute of Nuclear Energy Research (INER) had been successively decommissioned and decontaminated over the recent years. Since dismantling was a complex task, to achieve the main goal was minimization of radioactive waste production and required the set-up of procedures, criteria of free release, strict follow-up and traceability at all steps. This study gave an overview of the efforts on non-destructive assay (NDA) of relatively large volumes of waste and the sampling of contaminated waste with radiochemical analysis was utilized to determine the radionuclide vectors. The experiences of free release planning and measurement of a very low level radioactivity with high throughput for scrapped metal at the INER and the technical achievements in this research could offer a reference of decision-making by the competent authority.
Related JoVE Video
Ppp1r15 gene knockout reveals an essential role for translation initiation factor 2 alpha (eIF2alpha) dephosphorylation in mammalian development.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 01-30-2009
Show Abstract
Hide Abstract
Diverse cellular stress responses are linked to phosphorylation of serine 51 on the alpha subunit of translation initiation factor 2. The resultant attenuation of protein synthesis and activation of gene expression figure heavily in the adaptive response to stress, but dephosphorylation of eIF2(alphaP), which terminates signaling in this pathway, is less well understood. GADD34 and CReP, the products of the related mammalian genes Ppp1r15a and Ppp1r15b, can recruit phosphatase catalytic subunits of the PPP1 class to eIF2(alphaP), but the significance of their contribution to its dephosphorylation has not been explored systematically. Here we report that unlike Ppp1r15a mutant mice, which are superficially indistinguishable from wild type, Ppp1r15b(-/-) mouse embryos survive gestation but exhibit severe growth retardation and impaired erythropoiesis, and loss of both Ppp1r15 genes leads to early embryonic lethality. These loss-of-function phenotypes are rescued by a mutation, Eif2a(S51A), that prevents regulated phosphorylation of eIF2alpha. These findings reveal that the essential process of eIF2(alphaP) dephosphorylation is the predominant role of PPP1R15 proteins in mammalian development.
Related JoVE Video
The calibration and evaluation of a radioactive waste drum counting system.
Appl Radiat Isot
PUBLISHED: 01-24-2009
Show Abstract
Hide Abstract
The drum counting system was calibrated in this study. For (137)Cs, the counting efficiencies were around 14-1% when the density of the waste of the drum was changed from 0.15 to 2.3 g cm(-3). The effects of the background, hot spot, system linearity, sample density and weighing were also evaluated in this work. The combined standard uncertainty of the drum counting system for the (137)Cs in the density of 1g cm(-3) was around 12%. To verify the counting system, the drums containing radioactive solution were prepared by the NMI as the blind samples. A discrepancy below 15% was shown between the counting results and the reference values of the NMI.
Related JoVE Video
The behavior of rat tooth germ cells on poly(vinyl alcohol).
Acta Biomater
PUBLISHED: 01-13-2009
Show Abstract
Hide Abstract
The purpose of this study was to evaluate the behaviors of rat tooth germ (TG) cells cultured on poly(vinyl alcohol) (PVA). It was found that TG cells suspended and aggregated to form three-dimensional spheroids on PVA. Compared with traditional monolayered cells on tissue culture polystyrene, TG cell spheroids on PVA obviously increased the alkaline phosphatase activity, the degree of mineralization, and upregulated both osteopontin and dentin matrix protein 1 genes, regardless of the seeding density. Surprisingly, PVA appears to activate the alkaline phosphatase activity and mineralization effects on TG cell spheroids in the absence of a differentiation medium. Furthermore, the present study indicates that integrins may play an important role in the mineralization on TG cell spheroids by adding Arg-Gly-Asp (RGD) peptides. Therefore, the information presented here should help to clarify the role of PVA in the regulation of the mineralization, differentiation and integrin-mediation of TG cells.
Related JoVE Video
Computation of transient flow rates in passive pumping micro-fluidic systems.
Lab Chip
PUBLISHED: 01-07-2009
Show Abstract
Hide Abstract
Motion in micro-channels of passive flow micro-fluidic systems can be controlled by proper design and estimated by careful modeling. We report on methods to describe the flow rate as function of time in a passive pump driven micro-fluidic system. The model considers the surface energy present in small droplets, which prompts their shrinkage and induces flow. The droplet geometries are controlled by the micro-fluidic system geometry and hydrophilicity of the droplet channel contact area so that the chord of the droplets cross section is restrained as the fluid is pumped. The model uses interfacial thermodynamics and the Hagen-Poiseuille equation for calculating the flow rate in micro-channels. Existing analyses consider the theoretical relationships among sample volume and induced flow rate, surface energy of the drops at the entrance and exit ports, and the resistance to flow. This model provides more specific information on the influence of the experimental conditions in computations of the flow rate. The model was validated in four sets of experiments. Passive pumps with 1.8 mm diameter, hydrophobic or hydrophilic entry ports, 5.0 or 10.0 mm channel length, and 2.5 or 3.3 mm diameter reservoir ports provided initial flow rates between 85 nL s(-1) and 196 nL s(-1).
Related JoVE Video
Transverse and sagittal angulations of proximal segment after sagittal split and vertical ramus osteotomies and their influence on the stability of distal segment.
J. Formos. Med. Assoc.
Show Abstract
Hide Abstract
This study aimed at comparing the transverse and sagittal angulations of proximal segment after sagittal split ramus osteotomy (SSRO) and intraoral vertical ramus osteotomy (IVRO), and examining their influences on the stability of distal segment.
Related JoVE Video
Chemopreventive effects of pterostilbene on urethane-induced lung carcinogenesis in mice via the inhibition of EGFR-mediated pathways and the induction of apoptosis and autophagy.
J. Agric. Food Chem.
Show Abstract
Hide Abstract
Lung cancer is the most commonly diagnosed cancer and the leading cause of cancer deaths globally. Due to the lack of successful chemopreventive agents for lung cancer, there is an emerging need to evaluate new and effective agents for lung cancer prevention. Pterostilbene, a naturally occurring analogue of resveratrol, has been reported to be an effective chemopreventive agent against many cancers. The aim of this study is to investigate the chemopreventive effects of pterostilbene in urethane-induced murine lung tumors. Pretreatment with pterostilbene at 50 or 250 mg/kg significantly reduced tumor multiplicity by 26 and 49%, respectively. Pterostilbene also significantly inhibited tumor volume by 25 and 34% and decreased the tumor burden per mouse by 45 and 63%, respectively. The mechanisms by which pterostilbene suppresses lung tumorigenesis have been investigated in lung tissues and homogenates. The results indicate that the pterostilbene-mediated chemopreventive effects in vivo were a result of the inhibition of epidermal growth factor receptor (EGFR) and its downstream pathways, leading to retarded cell cycle progression, and of the induction of apoptosis and autophagy during urethane-induced lung tumorigenesis.
Related JoVE Video
Atrial fibrillation and atrial flutter: medical management.
Clin. Geriatr. Med.
Show Abstract
Hide Abstract
Atrial fibrillation (AF) and atrial flutter (AFL) are common cardiac arrhythmias in older adults. Medical management focuses on rate and rhythm control of AF and AFL to promote symptomatic relief and avoid tachycardia-mediated cardiomyopathy. Pharmacologic treatment of AF and AFL is especially challenging in the elderly because of the presence of comorbidities that may affect drug kinetics, and polypharmacy, which may lead to drug interactions. The potential for complications from medications and procedures required to achieve and maintain sinus rhythm must be carefully balanced against the benefits of therapy. This article reviews medical management of AF and AFL specifically relating to rate and rhythm control. The controversy of rate versus rhythm control is also discussed.
Related JoVE Video
Cloning, expression and characterization of CCL21 and CCL25 chemokines in zebrafish.
Dev. Comp. Immunol.
Show Abstract
Hide Abstract
Chemokines are a large group of proteins implicated in migration, activation, and differentiation of leukocytes. They are well-surveyed in mammals, but less is known in lower vertebrates about their spatiotemporal expressions and functions. From an evolutionary point of view, comparative analyses may provide some fundamental insights into these molecules. In mammals, CCL21 and CCL25 are crucial for thymocyte homing. Herein, we identified and cloned the zebrafish orthologues of CCL21 and CCL25, and analyzed their expression in embryos and adult fish by in situ hybridization. We found that CCL21 was expressed in the craniofacial region, pharynx, and blood vessels in embryos. In adult fish, CCL21 transcripts were located in the kidney, spinal cord, and blood cells. In contrast, expression of CCL25 was only detected in the thymus primordia in embryos. In adult fish, transcripts of CCL25 were maintained in the thymus, and they were also found in the brain and oocytes. Furthermore, we performed an antisense oligonucleotide experiment to evaluate the biological function of CCL25. Results showed that the recruitment of thymocytes was impeded by morpholino-mediated knockdown of CCL25, suggesting that CCL25 is essential for colonization of T-cells in the thymus in early development. Together, our results demonstrate the basic profiles of two CCL chemokines in zebrafish. The tissue-specific expression patterns may pave the way for further genetic dissection in this model organism.
Related JoVE Video
Establishment of clonal MIN-O transplant lines for molecular imaging via lentiviral transduction & in vitro culture.
PLoS ONE
Show Abstract
Hide Abstract
As the field of molecular imaging evolves and increasingly is asked to fill the discovery and validation space between basic science and clinical applications, careful consideration should be given to the models in which studies are conducted. The MIN-O mouse model series is an established in vivo model of human mammary precancer ductal carcinoma in situ with progression to invasive carcinoma. This series of transplant lines is propagated in vivo and experiments utilizing this model can be completed in non-engineered immune intact FVB/n wild type mice thereby modeling the tumor microenvironment with biological relevance superior to traditional tumor cell xenografts. Unfortunately, the same qualities that make this and many other transplant lines more biologically relevant than standard cell lines for molecular imaging studies present a significant obstacle as somatic genetic re-engineering modifications common to many imaging applications can be technically challenging. Here, we describe a protocol for the efficient lentiviral transduction of cell slurries derived from precancerous MIN-O lesions, in vitro culture of "MIN-O-spheres" derived from single cell clones, and the subsequent transplantation of these spheres to produce transduced sublines suitable for optical imaging applications. These lines retain the physiologic and pathologic properties, including multilineage differentiation, and complex microanatomic interaction with the host stroma characteristic of the MIN-O model. We also present the in vivo imaging and immunohistochemical analysis of serial transplantation of one such subline and detail the progressive multifocal loss of the transgene in successive generations.
Related JoVE Video

What is Visualize?

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

How does it work?

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

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

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