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Find video protocols related to scientific articles indexed in Pubmed.
Immunoglobulin E plays an immunoregulatory role in lupus.
J. Exp. Med.
PUBLISHED: 09-29-2014
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The (patho)physiological role of IgE in nonallergic inflammatory diseases is not well understood. Here, we explored the effect of IgE deficiency on the inflammatory response in Fc?RIIB-deficient mice as well as in mice carrying both a deletion of Fc?RIIB and the chromosomal translocation of Y-linked autoimmune acceleration (Yaa) that hastens and results in a more aggressive lupuslike disease in these mice. The findings show that deficiency of IgE delays disease development and severity as demonstrated by reduced autoantibody production and amelioration of organ pathologies. This was associated with decreased numbers of plasma cells and reduced levels of IgG2b and IgG3. Unexpectedly, the loss of IgE also caused a striking decrease of immune cell infiltration in secondary lymphoid organs with a marked effect on the presence of dendritic cells, monocytes, neutrophils, and eosinophils in these organs and decreased activation of basophils. The presence of autoreactive IgE in human systemic lupus erythematosus subjects was also associated with increased basophil activation and enhanced disease activity. These findings argue that IgE facilitates the amplification of autoimmune inflammation.
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[Radical nephrectomy and caval tumor thrombectomy with cardiopulmonary bypass combined with right anterior minithoracotomy;report of a case].
Kyobu Geka
PUBLISHED: 09-10-2014
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Renal cell carcinoma is a tumor with a distinct feature that it can invade the renal vein and grow intravascularly extending to the inferior vena cava (IVC). We herein report a case of a 71-year-old female who presented with a neoplasm that involved the right kidney and an intra-IVC tumor thrombus. We performed radical nephrectomy and tumor thrombectomy under cardiopulmonary bypass through aright anterior minithoracotomy. The patient was discharged on the 13th day after the surgery without any complication, and is currently in good health at 7 months after the operation.
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Rethinking the role of immunoglobulin e and its high-affinity receptor: new insights into allergy and beyond.
Int. Arch. Allergy Immunol.
PUBLISHED: 09-09-2014
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Immunoglobulin E (IgE) and its high-affinity receptor (Fc?RI) are well-known participants in the allergic response. The interaction of allergens with Fc?RI-bound IgE antibodies is an essential step in mast cell/basophil activation and the subsequent release of allergic mediators. It is known that the affinity of the interaction between an IgE antibody and an allergen may differ, raising the question of whether Fc?RI can decipher these differences. If so, do the cellular and physiological outcomes vary? Are the molecular mechanisms initiated by Fc?RI similarly under low- or high-affinity interactions? Could the resulting inflammatory response differ? Recent discoveries summarized herein are beginning to shed new light on these important questions. What we have learned from them is that IgE and Fc?RI form a complex regulatory network influencing the inflammatory response in allergy and beyond. © 2014 S. Karger AG, Basel.
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Phosphorylation of Smad2/3 at Specific Linker Threonine Indicates Slow-Cycling Intestinal Stem-Like Cells Before Reentry to Cell Cycle.
Dig. Dis. Sci.
PUBLISHED: 09-04-2014
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Quiescent (slow-cycling) and active (rapid-cycling) stem cells are demonstrated in small intestines. We have identified significant expression of Smad2/3, phosphorylated at specific linker threonine residues (pSmad2/3L-Thr), in murine stomach, and suggested these cells are epithelial stem cells.
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Prognosis-related factors concerning oral and general conditions for homebound older adults in Japan.
Geriatr Gerontol Int
PUBLISHED: 08-03-2014
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The present study examined the relationship between oral function, such as eating/swallowing, and life prognosis among a homebound elderly population, considering physical and mental function.
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Orai-2 is localized on secretory granules and regulates antigen-evoked Ca²? mobilization and exocytosis in mast cells.
Biochem. Biophys. Res. Commun.
PUBLISHED: 06-23-2014
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The increase in intracellular Ca(2+) through the Ca(2+) channel is an indispensable step for the secretion of inflammatory mediators by mast cells. It was recently reported that Orai-1 is responsible for the Ca(2+) influx that is activated by depletion of stored Ca(2+). There are three isoforms of Orai: Orai-1, Orai-2, and Orai-3; however, isoforms other than Orai-1 are poorly understood. We found that Orai-2 is expressed and localized on secretory granules in RBL-2H3. Ca(2+) release from Ca(2+) store, induced by antigen stimulation, was significantly attenuated by knockdown of Orai-2, while that induced by thapsigargin was not affected. Furthermore, exocytotic release induced by antigen stimulation was inhibited in knockdown cells. This observation suggests a new role of Orai isoforms in secretory cells.
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Gastric diffuse large B-cell lymphoma cured with Helicobacter pylori eradication regardless of whether it contains features of MALT lymphoma.
Intern. Med.
PUBLISHED: 04-04-2014
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A 66-year-old patient was diagnosed with primary gastric B-cell lymphoma. The pathological findings were consistent with diffuse large B-cell lymphoma (DLBCL); however, a small area showed features of mucosa-associated lymphoid tissue (MALT) lymphoma. Biopsy specimens were referred to two other pathologists, both of whom diagnosed the case as pure DLBCL, denying the area of MALT lymphoma. As the lymphoma was limited to the submucosal layer and the patient's general condition was excellent, eradication of Helicobacter pylori was selected as the initial treatment. The lymphoma completely disappeared three months after the eradication treatment, and complete remission has been maintained for nearly two years.
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[A review of the complications and candida colonization associated with voice rehabilitation using a voice prosthesis].
Nippon Jibiinkoka Gakkai Kaiho
PUBLISHED: 03-08-2014
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Between December 2004 and December 2011, we have used the indwelling voice prosthesis for voice rehabilitation in 28 patients after total laryngectomy in our department. To clarify both the complications and safety of voice reconstruction using a voice prosthesis, and the occurrence of candida colonization, we conducted a retrospective study with a review of the Japanese literature. Twenty-six patients who were observed over a period of more than six months at our hospital were enrolled in this study. We examined the interval of prosthesis replacement, types and frequency of complications, candida colonization, and clinical outcomes. The median follow-up time postoperatively was 28.8 months (range 8.1-95.7). The average interval of prosthesis replacement was 147 days (4.9 months). Complications occurred in 14 patients (54%), and 6 patients (23%) of whom were hospitalized. The main complications were periprosthetic leakage, increased granulation around the tracheoesophageal shunt and stenosis of the trachea stoma. There were no significant differences in the frequency of complications in the background factor of the patients as far as age (p = 0.495), radiation therapy (p = 0.686) or reconstruction time (p = 0.257) were concerned. Candida species was detected in 81% of the specimens which we submitted to a culture test and confirmed the pseudohyphae which comfirmed the pathogenicity from the histopathological examination. Moreover, radiation therapy was significantly associated with the detection of candida (p = 0.004). Permanent closure of the tracheoesophageal shunt for periprosthetic leakage was required in one patient, but we were able to deal with the other complications. No patient experienced any life-threatening complications and all are safely using the prostheses. It has been reported that complications will occur over the long-term, thus careful follow-up is necessary.
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Pseudoaneurysm of the ascending aorta 31 years after mitral valve replacement.
Asian Cardiovasc Thorac Ann
PUBLISHED: 03-04-2014
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We describe a rare case of an ascending aortic pseudoaneurysm 31 years after mitral valve replacement with a Björk-Shiley mechanical valve. The aneurysm presumably expanded gradually during the years following surgery. As the valve was functioning normally, it was left in situ while the ascending aorta was replaced. This report provides valuable information regarding the long-term nature of this patient's pseudoaneurysm, and the long-term durability of the Björk-Shiley spherical valve in the mitral position.
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Aortic arch surgery in octogenarians: is it justified?
Eur J Cardiothorac Surg
PUBLISHED: 02-28-2014
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Elderly patients are sometimes denied aortic arch surgery because of the perception of poor outcomes and an unacceptable quality of life (QOL). In this study, we evaluated the early clinical outcomes, long-term survival and QOL following surgical treatment for aortic arch disease in octogenarian patients.
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Kidney-selective gene transfection using anionic bubble lipopolyplexes with renal ultrasound irradiation in mice.
Nanomedicine
PUBLISHED: 02-14-2014
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This study assessed the ability of a new ultrasound (US) responsive gene delivery carrier, bubble lipopolyplexes, to deliver genes to the kidneys. The bubble lipopolyplexes showed highly selective gene expression in kidney tubules, but only after renal irradiation with US. These bubble lipopolyplexes, however, did not increase the expression of biomarkers of kidney injury, including blood urea nitrogen, serum creatinine, kidney injury molecule-1 mRNA, and clusterin mRNA, or induce any histopathological abnormalities in the kidney. Furthermore, pDNA containing CMV early enhancer/chicken beta-actin promoter prolonged gene expression by the bubble lipopolyplexes in the kidney for 42days. This novel renal gene delivery method, in which transfection of bubble lipopolyplexes was followed by US irradiation of the kidneys, resulting in cell-selective, high, and long-term gene expression without renal injury in mice, may have future applications in patient treatment.
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Lung abscess without sepsis in a patient with diabetes with refractory episodes of spontaneous hypoglycemia: a case report and review of the literature.
J Med Case Rep
PUBLISHED: 02-13-2014
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Hypoglycemia is a cause of considerable morbidity. Although hypoglycemia has been documented in the setting of septic shock and has been associated with higher mortality, hypoglycemia in infection without sepsis has not been reported in the literature.
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Mediastinal schwannoma diagnosed preoperatively as a cyst.
Tex Heart Inst J
PUBLISHED: 02-12-2014
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Mediastinal schwannomas are sometimes diagnosed as pericardial or bronchogenic cysts, if cystic degeneration is extensive. When mediastinal schwannomas are not diagnosed as primary cardiac tumors, the use of cardiopulmonary bypass in their resection appears to be infrequent. We report the case of a 48-year-old woman who presented with symptoms from a suspected intrapericardial cyst. Multiple diagnostic images revealed a large mass, potentially a proteinaceous or hemorrhagic cyst, in the transverse sinus behind the ascending aorta and against the left main trunk. After complete resection with the use of cardiopulmonary bypass, the mass was identified as a benign extracardiac schwannoma. More than 3 years postoperatively, the patient had no relevant symptoms. We discuss the preoperative diagnosis, the method of resection, and our broad strategy for dealing with such a case.
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Molecular editing of cellular responses by the high-affinity receptor for IgE.
Science
PUBLISHED: 02-06-2014
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Cellular responses elicited by cell surface receptors differ according to stimulus strength. We investigated how the high-affinity receptor for immunoglobulin E (IgE) modulates the response of mast cells to a high- or low-affinity stimulus. Both high- and low-affinity stimuli elicited similar receptor phosphorylation; however, differences were observed in receptor cluster size, mobility, distribution, and the cells' effector responses. Low-affinity stimulation increased receptor association with the Src family kinase Fgr and shifted signals from the adapter LAT1 to the related adapter LAT2. LAT1-dependent calcium signals required for mast cell degranulation were dampened, but the role of LAT2 in chemokine production was enhanced, altering immune cell recruitment at the site of inflammation. These findings uncover how receptor discrimination of stimulus strength can be interpreted as distinct in vivo outcomes.
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[Base in four types of lidocaine preparation (formulated in hospital)].
Yakugaku Zasshi
PUBLISHED: 02-05-2014
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PL cream (combination of lidocaine and procaine) was launched on the market in April 2012 in Japan. We investigated differences in the anesthetic effect by employing two types of base: Carbopol and methylcellulose. Electron microscopy showed a distinct difference in appearance: densely-scattered, fine particles for Carbopol and sparse, large particles for methylcellulose. Accordingly, the extensibility of the cream was significantly greater at 4 and 25 degrees centigrade for methylcellulose, but was greater at 34 degrees centigrade for Carbopol. The steady flow viscosity (1 s(-1)) was greater for the Carbopol than methylcellulose base. The difference in the cutaneous permeability between the two bases increased over time: the methylcellulose base was removed at 90 min after application and, 30 min later, showed a significant difference. These results suggest that the methylcellulose base has a superior anesthetic effect in clinical settings.
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Deregulation of pancreas-specific oxidoreductin ERO1? in the pathogenesis of diabetes mellitus.
Mol. Cell. Biol.
PUBLISHED: 01-27-2014
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A growing body of evidence has underlined the significance of endoplasmic reticulum (ER) stress in the pathogenesis of diabetes mellitus. ER oxidoreductin 1? (ERO1?) is a pancreas-specific disulfide oxidase that is known to be upregulated in response to ER stress and to promote protein folding in pancreatic ? cells. It has recently been demonstrated that ERO1? promotes insulin biogenesis in ? cells and thus contributes to physiological glucose homeostasis, though it is unknown if ERO1? is involved in the pathogenesis of diabetes mellitus. Here we show that in diabetic model mice, ERO1? expression is paradoxically decreased in ? cells despite the indications of increased ER stress. However, overexpression of ERO1? in ? cells led to the upregulation of unfolded protein response genes and markedly enlarged ER lumens, indicating that ERO1? overexpression caused ER stress in the ? cells. Insulin contents were decreased in the ? cells that overexpressed ERO1?, leading to impaired insulin secretion in response to glucose stimulation. These data indicate the importance of the fine-tuning of the ER redox state, the disturbance of which would compromise the function of ? cells in insulin synthesis and thus contribute to the pathogenesis of diabetes mellitus.
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Bubble liposomes and ultrasound exposure improve localized morpholino oligomer delivery into the skeletal muscles of dystrophic mdx mice.
Mol. Pharm.
PUBLISHED: 01-27-2014
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Duchenne muscular dystrophy (DMD) is a genetic disorder that is caused by mutations in the DMD gene that lead to an absence of functional protein. The mdx dystrophic mouse contains a nonsense mutation in exon 23 of the dystrophin gene; a phosphorodiamidate morpholino oligomer (PMO) designed to skip this mutated exon in the mRNA induces dystrophin expression. However, an efficient PMO delivery method is needed to improve treatment strategies for DMD. We previously developed polyethylene glycol (PEG)-modified liposomes (Bubble liposomes) that entrap ultrasound contrast gas and demonstrated that the combination of Bubble liposomes with ultrasound exposure is an effective gene delivery tool in vitro and in vivo. In this study, to evaluate the ability of Bubble liposomes as a PMO delivery tool, we tested the potency of the Bubble liposomes combined with ultrasound exposure to boost the delivery of PMO and increase the skipping of the mutated exon in the mdx mouse. The results indicated that the combination of Bubble liposomes and ultrasound exposure increased the uptake of the PMO targeting a nonsense mutation in exon 23 of the dystrophin gene and consequently increased the PMO-mediated exon-skipping efficiency compared with PMO injection alone, leading to significantly enhanced dystrophin expression. This increased efficiency indicated the potential of the combination of Bubble liposomes with ultrasound exposure to enhance PMO delivery for treating DMD. Thus, this ultrasound-mediated Bubble liposome technique may provide an effective, noninvasive, nonviral method for PMO therapy for DMD muscle as well as for other muscular dystrophies.
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Haemodynamic unloading increases the survival and affects the differentiation of cardiac stem cells after implantation into an infarcted heart.
Eur J Cardiothorac Surg
PUBLISHED: 01-23-2014
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It has been anticipated that stem cell therapy is capable of repairing an injured heart but is currently limited by its marginal efficacy. We believe that mechanical stress due to haemodynamic loading may negate the therapeutic potency of stem cells and therefore investigated how haemodynamic unloading affects the survival and differentiation of stem cells after implantation into an infarcted heart.
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Systemic delivery of miR-126 by miRNA-loaded Bubble liposomes for the treatment of hindlimb ischemia.
Sci Rep
PUBLISHED: 01-09-2014
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Currently, micro RNA (miRNA) is considered an attractive target for therapeutic intervention. A significant obstacle to the miRNA-based treatments is the efficient delivery of miRNA to the target tissue. We have developed polyethylene glycol-modified liposomes (Bubble liposomes (BLs)) that entrap ultrasound (US) contrast gas and can serve as both plasmid DNA (pDNA) or small interfering RNA (siRNA) carriers and US contrast agents. In this study, we investigated the usability of miRNA-loaded BLs (mi-BLs) using a hindlimb ischemia model and miR-126. It has been reported that miR-126 promotes angiogenesis via the inhibition of negative regulators of VEGF signaling. We demonstrated that mi-BLs could be detected using diagnostic US and that mi-BLs with therapeutic US could deliver miR-126 to an ischemic hindlimb, leading to the induction of angiogenic factors and the improvement of blood flow. These results suggest that combining mi-BLs with US may be useful for US imaging and miRNA delivery.
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Combination of bubble liposomes and high-intensity focused ultrasound (HIFU) enhanced antitumor effect by tumor ablation.
Biol. Pharm. Bull.
PUBLISHED: 01-07-2014
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Ultrasound (US) is used in the clinical setting not only for diagnosis but also for therapy. As a therapeutic US technique, high-intensity focused ultrasound (HIFU) can be applied to treat cancer in a clinical setting. Microbubbles increased temperature and improved the low therapeutic efficiency under HIFU; however, microbubbles have room for improvement in size, stability, and targeting ability. To solve these issues, we reported that "Bubble liposomes" (BLs) containing the US imaging gas (perfluoropropane gas) liposomes were suitable for ultrasound imaging and gene delivery. In this study, we examined whether BLs and HIFU could enhance the ablation area of the tumor and the antitumor effect. First, we histologically analyzed the tumor after BLs and HIFU. The ablation area of the treatment of BLs and HIFU was broader than that of HIFU alone. Next, we monitored the temperature of the tumor, and examined the antitumor effect. The temperature increase with BLs and HIFU treatment was faster and higher than that with HIFU alone. Moreover, treatment with BLs and HIFU enhanced the antitumor effect, which was better than with HIFU alone. Thus, the combination of BLs and HIFU could be efficacious for cancer therapy.
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Mortality and morbidity after total aortic arch replacement.
Ann. Thorac. Surg.
PUBLISHED: 01-06-2014
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Total aortic arch replacement is associated with considerable mortality and morbidity. Although operative death is the most extreme adverse clinical end point, postoperative morbidity can also be devastating for survivors.
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[Apico-aortic Valved Conduit for Aortic Stenosis in Patients Who Could not Tolerate Median Sternotomy].
Kyobu Geka
PUBLISHED: 12-11-2013
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This report presents 3 cases treated with an apico-aortic valved conduit. Cases 1, 2:A 67-year-old female patient and a 60-year-old male patient what had undergone coronary artery bypass grafting were admitted to our hospital for severe aortic stenosis. Computed tomography showed a severe calcified ascending aorta, and coronary angiography revealed patent bypass graft. Case 3:A 71-year-old male patient that had esophagectomy with retrosternal colonic interposition for esophagus cancer after distal gastrectomy. In addition, he had experienced anterior mediastinal drainagic therapy for anastomotic leak. All 3 patients were treated by implantation of an apico-aortic valved conduit. Operation:This procedure was performed through the 5th intercostal space under a beating heart with cardiopulmonary bypass. Result:Postoperative courses were uneventful. All patients are still alive without procedure-related events. Conclusion:This surgical procedure can be an effective alternative when conventional aortic valve replacement cannot be performed for aortic stenosis patients.
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Munc18-2 and syntaxin 3 control distinct essential steps in mast cell degranulation.
J. Immunol.
PUBLISHED: 12-09-2013
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Mast cell degranulation requires N-ethylmaleimide-sensitive factor attachment protein receptors (SNARE) and mammalian uncoordinated18 (Munc18) fusion accessory proteins for membrane fusion. However, it is still unknown how their interaction supports fusion. In this study, we found that small interfering RNA-mediated silencing of the isoform Munc18-2 in mast cells inhibits cytoplasmic secretory granule (SG) release but not CCL2 chemokine secretion. Silencing of its SNARE-binding partner syntaxin 3 (STX3) also markedly inhibited degranulation, whereas combined knockdown produced an additive inhibitory effect. Strikingly, while Munc18-2 silencing impaired SG translocation, silencing of STX3 inhibited fusion, demonstrating unique roles of each protein. Immunogold studies showed that both Munc18-2 and STX3 are located on the granule surface, but also within the granule matrix and in small nocodazole-sensitive clusters of the cytoskeletal meshwork surrounding SG. After stimulation, clusters containing both effectors were detected at fusion sites. In resting cells, Munc18-2, but not STX3, interacted with tubulin. This interaction was sensitive to nocodazole treatment and decreased after stimulation. Our results indicate that Munc18-2 dynamically couples the membrane fusion machinery to the microtubule cytoskeleton and demonstrate that Munc18-2 and STX3 perform distinct, but complementary, functions to support, respectively, SG translocation and membrane fusion in mast cells.
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Application of Ultrasound-enhanced Gene and Drug Delivery to the Ocular Tissue.
Yakugaku Zasshi
PUBLISHED: 12-03-2013
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  Visual images provide an immensely rich source of information about the external world. Eye has characteristic structure sensory cells are arranged along the eye wall, and is filled inside with vitreous body. In recent years, intravitreal injection of anti-vascular endothelial growth factor (VEGF) agent had widely spread, and numerous number of patients who suffered ocular angiogenic disease such as diabetic retinopathy, age-related macular degeneration and retinal vascular occlusion for the disease, were treated and spared the blindness. Vitreous cavity was regarded as reservoir of drug, intravitreal injection is thought a sort of drug delivery. However, with regard to the administration of a selective drug deliver, it has not yet been solved. Our aim is to establish a new method of gene transfer, drug delivery using low-energy ultrasound to the eye, to date, we confirmed drug and gene deliver to the ocular tissue such as cornea, conjunctiva and retina with high efficiency. In addition, tissue damage was minimal. We have also shown that ultrasound irradiation with combination of a microbubbles or bubble liposome could be introduced drug and gene more effectively. Based on these knowledge, we will focus on development of a new device for intraocular ultrasound exposure and potential for therapeutic application of ultrasound to humans retinal disease such as retinal artery obstruction.
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Novel dianostics and therapeutics with ultrasound technologies and nanotechnologies.
Yakugaku Zasshi
PUBLISHED: 12-03-2013
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  Ultrasound is a good tool for theranostics due to have multi-potency both of diagnostics with sonography and therapeutics with high intensity focused ultrasound (HIFU). In addition, microbubbles and nanobubbles are utilized as not only contrast imaging agent but also enhancer of drug and gene delivery by combination of ultrasound. Recently, we developed novel liposomal nanobubbles (Bubble liposomes) which were containing perfluoropropane. Bubble liposomes induced jet stream by low intensity ultrasound exposure and resulted in enhancing permeability of cell membrane. This phenomenon has been utilized as driving force for drug and gene delivery. On the other hand, the combination of Bubble liposomes and high intensity ultrasound induces strong jet stream and increase temperature. This condition can directly damage to tumor cells, we are applying this for cancer therapy. Therefore, their combination has potency for various cancer therapies such as gene therapy, immunotherapy and hyperthermia. In this review, we discuss about cancer therapy by the combination of Bubble liposomes and ultrasound.
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Effects of an Angiotensin II Receptor Blocker on the Impaired Function of Endothelial Progenitor Cells in Patients With Essential Hypertension.
Am. J. Hypertens.
PUBLISHED: 11-07-2013
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Endothelial progenitor cells (EPCs) induce neovascularization and repair vascular damage. We have demonstrated that EPC function is impaired in hypertensive rats with increases in oxidative stress and that angiotensin II receptor blockers improved the impaired function of EPCs. In this study, we investigated basal EPC functions in normotensive control subjects and patients with essential hypertension and the effect of losartan on EPC function in hypertensive patients.
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Development of anionic bubble lipopolyplexes for efficient and safe gene transfection with ultrasound exposure in mice.
J Control Release
PUBLISHED: 09-29-2013
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Anionic bubble lipopolyplexes have been developed as anionic ultrasound (US)-responsive gene delivery carriers with biocompatible compounds for efficient and safe transfection in mice. The particles of the anionic bubble lipopolyplexes were approximately 450-600nm with an anionic surface charge. In the absence of US exposure, the bubble lipopolyplexes showed extremely low gene expression in the human vascular endothelial cell line EAhy926. The anionic bubble lipopolyplexes, however, delivered pDNA into cells without endocytosis and showed markedly high gene expression following US exposure. The anionic bubble lipopolyplexes showed little cytotoxicity in EAhy926 cells and little aggregation with erythrocytes. Following intravenous administration into mice, the anionic bubble lipopolyplexes showed high levels of gene expression in the liver, kidney, and spleen only after US exposure to the abdominal area. The level of gene expression in liver non-parenchymal cells was significantly higher than that in parenchymal cells. In addition, the anionic bubble lipopolyplexes did not show any severe hepatic toxicity and did not enhance the production of proinflammatory cytokines. Overall, we have succeeded in preparing anionic bubble lipopolyplexes for efficient and safe transfection with US exposure in mice.
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Bioengineered trachea using autologous chondrocytes for regeneration of tracheal cartilage in a rabbit model.
Laryngoscope
PUBLISHED: 07-02-2013
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In this study, a bioengineered trachea composed of autologous chondrocytes was developed, and its effect on cartilaginous regeneration was evaluated by implantation into tracheal defects in rabbits.
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Predictors of late aortic events after Stanford type B acute aortic dissection.
J. Thorac. Cardiovasc. Surg.
PUBLISHED: 04-16-2013
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Patients with Stanford type B acute aortic dissection usually receive medical treatment during the acute phase. The present study aimed to elucidate the factors predicting late aortic events in patients treated conservatively for acute type B dissections.
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Aberrant methylation of microRNA-34b/c is a predictive marker of metachronous gastric cancer risk.
J. Gastroenterol.
PUBLISHED: 04-08-2013
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Metachronous gastric cancer (GC) can develop after endoscopic resection of GC and cannot be predicted based on clinical signature. Aberrant DNA methylation in noncancerous gastric mucosa is strongly implicated in gastric carcinogenesis and could be a useful biomarker of GC risk. We evaluated the clinical utility of DNA methylation as a biomarker of metachronous GC risk.
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Reduction of the cholesterol sensor SCAP in the brains of mice causes impaired synaptic transmission and altered cognitive function.
PLoS Biol.
PUBLISHED: 04-01-2013
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The sterol sensor SCAP is a key regulator of SREBP-2, the major transcription factor controlling cholesterol synthesis. Recently, we showed that there is a global down-regulation of cholesterol synthetic genes, as well as SREBP-2, in the brains of diabetic mice, leading to a reduction of cholesterol synthesis. We now show that in mouse models of type 1 and type 2 diabetes, this is, in part, the result of a decrease of SCAP. Homozygous disruption of the Scap gene in the brains of mice causes perinatal lethality associated with microcephaly and gliosis. Mice with haploinsufficiency of Scap in the brain show a 60% reduction of SCAP protein and ~30% reduction in brain cholesterol synthesis, similar to what is observed in diabetic mice. This results in impaired synaptic transmission, as measured by decreased paired pulse facilitation and long-term potentiation, and is associated with behavioral and cognitive changes. Thus, reduction of SCAP and the consequent suppression of cholesterol synthesis in the brain may play an important role in the increased rates of cognitive decline and Alzheimer disease observed in diabetic states.
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Technique for using pedicled latissimus dorsi muscle flaps to wrap prosthetic grafts in an infected thoracic aorta.
Ann Vasc Surg
PUBLISHED: 03-30-2013
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In this report we describe a special method of using a pedicled latissimus dorsi muscle flap for a mycotic aneurysm. The method involves wrapping the flap around a prosthetic graft. Using this method, an in situ graft, which replaces an aneurysm, can help to avoid recurrence of local infection.
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Resection of an intrapericardial lipoma with nonsustained ventricular tachycardia.
J Card Surg
PUBLISHED: 03-28-2013
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A 35-year-old female presented with nonsustained ventricular tachycardia (NSVT) and a large intrapericardial mass. The mass was attached to the left ventricular wall with a broad stalk. On histopathology, the tumor was diagnosed as a lipoma. Postoperatively, the NSVT disappeared.
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Malignant solitary fibrous tumor of the left ventricular epicardium.
Ann. Thorac. Surg.
PUBLISHED: 03-26-2013
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Reports describing solitary fibrous tumors of the pericardium are rare. Surgical treatment was performed on a 49-year-old woman with a large pericardial mass. The mass was attached to the left ventricular wall with a broad stalk and was free of the parietal pericardium. It was apparent macroscopically that the tumor had invaded the left ventricular muscle. On histopathology, the tumor was diagnosed as a solitary fibrous tumor with low-grade malignancy.
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Suppression of murine collagen-induced arthritis by vaccination of synovial vascular endothelial cells.
Life Sci.
PUBLISHED: 03-21-2013
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Endothelial cells (ECs) lining the lumina of blood vessels are involved in leukocyte extravasation underlying inflammatory states, such as rheumatoid arthritis (RA). The rheumatoid pannus, the site of inflammation and joint destruction in the rheumatoid synovium, relies on the development of neovascular vessels to sustain its growth. We studied a method to selectively target and destroy new synovial blood vessels by vaccination with synovial EC.
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Simplicity, skills, and pitfalls of ascending aortic cannulation for type A aortic dissection.
J Cardiothorac Surg
PUBLISHED: 03-17-2013
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Ascending aortic cannulation for an antegrade central perfusion during surgery for type A aortic dissection is simple and can be performed within median sternotomy. This cannulation is performed routinely without problems in our hospital. Using our experience, the skills and pitfalls were clarified to make this challenging procedure successful.
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Ratio of preoperative atrial natriuretic peptide to brain natriuretic peptide predicts the outcome of the maze procedure in mitral valve disease.
J Cardiothorac Surg
PUBLISHED: 02-27-2013
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Although the maze procedure is an established surgical treatment for eliminating atrial fibrillation (AF), its efficacy in patients with mitral valve disease has remained unsatisfactory. A useful predictive marker for the outcome of the maze procedure is needed. The aim of this study was to investigate whether the preoperative ratio of atrial natriuretic peptide (ANP) to brain natriuretic peptide (BNP) reflects atrial fibrosis and can be used to predict the maze procedure outcome in patients with mitral valve disease.
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Transapical extirpation of a left ventricular thrombus in Takotsubo cardiomyopathy.
J Cardiothorac Surg
PUBLISHED: 02-01-2013
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A 58-year-old Japanese female was referred to our hospital. Although the electrocardiogram showed ST elevation, coronary angiography showed intact coronary artery. We diagnosed Takotsubo cardiomyopathy and a left ventricular thrombus. Anticoagulation was administered; however, the left ventricular thrombus had become mobile and protrusive. We extirpated the left ventricular thrombus via trans-apical approach. Left ventricular thrombus is rare in Takotsubo cardiomyopathy, but these patients are at a higher risk of thromboembolism, especially if the thrombi are mobile and protruding.
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Ultrasound-mediated gene delivery systems by AG73-modified bubble liposomes.
Biopolymers
PUBLISHED: 02-01-2013
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Targeted gene delivery to neovascular vessels in tumors is considered a promising strategy for cancer therapy. We previously reported that "Bubble liposomes" (BLs), which are ultrasound (US) imaging gas-encapsulating liposomes, were suitable for US imaging and gene delivery. When BLs are exposed to US, the bubble is destroyed, creating a jet stream by cavitation, and resulting in the instantaneous ejection of extracellular plasmid DNA (pDNA) or other nucleic acids into the cytosol. We developed AG73 peptide-modified Bubble liposomes (AG73-BL) as a targeted US contrast agent, which was designed to attach to neovascular tumor vessels and to allow specific US detection of angiogenesis (Negishi et al., Biomaterials 2013, 34, 501-507). In this study, to evaluate the effectiveness of AG73-BL as a gene delivery tool for neovascular vessels, we examined the gene transfection efficiency of AG73-BL with US exposure in primary human endothelial cells (HUVEC). The transfection efficiency was significantly enhanced if the AG73-BL attached to the HUVEC was exposed to US compared to the BL-modified with no peptide or scrambled peptide. In addition, the cell viability was greater than 80% after transfection with AG73-BL. These results suggested that after the destruction of the AG73-BL with US exposure, a cavitation could be effectively induced by the US exposure against AG73-BL binding to the cell surface of the HUVEC, and the subsequent gene delivery into cells could be enhanced. Thus, AG73-BL may be useful for gene delivery as well as for US imaging of neovascular vessels. © 2013 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 100: 402-407, 2013.
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Usage of sphingosine kinase isoforms in mast cells is species and/or cell type determined.
J. Immunol.
PUBLISHED: 01-28-2013
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Fc?RI engagement in mast cells (MCs) induces the activation of two distinct sphingosine kinase isoforms (SphK1 and SphK2) to produce sphingosine-1-phosphate, a mediator essential for MC responses. Whereas embryonic-derived SphK2-null MCs showed impaired responses to Ag, RNA silencing studies on other MC types indicated a dominant role for SphK1. Given the known functional heterogeneity of MCs, we explored whether the reported differences in SphK1 or SphK2 usage could be reflective of phenotypic differences between MC populations. Using lentiviral-based short hairpin RNA to silence SphK1 or SphK2, we found that SphK2 is required for murine MC degranulation, calcium mobilization, and cytokine and leukotriene production, irrespective of the tissue from which the MC progenitors were derived, the stage of MC granule maturity, or the conditions used for differentiation. This finding was consistent with the lack of a full allergic response in SphK2-null mice challenged to undergo passive cutaneous anaphylaxis. A redundant role for both SphKs was uncovered, however, in chemotaxis toward Ag in all MC types tested and in TNF-? production in certain MC types. In contrast, human MC responses were dependent only on SphK1, associating with a more robust expression of this isoform and a more varied representation of SphK variants relative to murine MCs. The findings show that the function of SphK1 and SphK2 can be interchangeable in MCs; however, an important determinant of SphK isoform usage is the species of origin and an influencing factor, the tissue from which MCs may be derived and/or their differentiation state.
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Arch translocation and the intra-arch elephant-trunk technique with collared graft for extended chronic dissecting aortic aneurysm.
J Cardiothorac Surg
PUBLISHED: 01-22-2013
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Management of extensive, chronic, dissecting aortic aneurysms after prior repair of the ascending aorta presents a technical challenge for surgeons. A symptomatic 64-year-old patient was admitted for elective surgical repair of an aortic annular dilatation, causing severe aortic regurgitation, and a Crawford type II extended thoracoabdominal aneurysm, 4?years after he underwent primary repair of an acute aortic dissection. The aorta was diffusely dilated, and there were no sites beyond the distal aortic arch where anastomosis could be performed. We successfully performed total aortic replacement with a 2-stage strategy, using an arch translocation technique and an intra-arch elephant-trunk technique.
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pDNA-loaded Bubble liposomes as potential ultrasound imaging and gene delivery agents.
Biomaterials
PUBLISHED: 01-21-2013
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We have developed polyethyleneglycol (PEG)-modified liposomes (Bubble liposomes; BLs) that entrap ultrasound (US) contrast gas, and we have reported that the combination of BLs and US exposure was an effective tool for delivering pDNA and siRNA in vitro and in vivo. In this study, we prepared pDNA-loaded BLs using three types of cationic lipids to enhance the US imaging effect and the transfection efficiency via systemic injection. We investigated the US imaging abilities of these BLs, their protective effects on pDNA from serum component, and their transfection effects in vitro and in vivo. As a result, we demonstrated that the US imaging ability and transfection effect varied with lipid component and that p-BLs containing DSDAP could be the most stable and effective tool the among three types of p-BLs. Indeed, in ischemic muscle, p-BLs containing DSDAP could be detected using diagnostic US and could deliver bFGF-expressing pDNA using therapeutic US, leading to the induction of angiogenic factors and the improvement of blood flow. These results suggest that combining p-BLs with US exposure may be useful for stable US imaging and efficient gene delivery and may lead to the establishment of a theranostic approach, which is a combination of disease diagnosis and therapy.
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The Association of Calf Circumference with Resting Pulse Rate in Community-dwelling Healthy Elderly Women -Pilot Study-.
J Phys Ther Sci
PUBLISHED: 01-09-2013
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[Purpose] High resting blood pressure and heart rate are associated with the risk of cardiovascular events. In patients with decreasing amounts of slow twitch muscle fiber, hypertension may develop and resting heart rate may increase. Measurement of the amount of slow twitch muscle fiber and capillary density using muscle biopsy is inconvenient and expensive. Therefore, a better screening test is required to determine these parameters for prevention of cardiovascular events. In this study, relationships among calf circumference, resting blood pressure, and pulse rate in the soleus muscle were investigated. [Subjects] Healthy elderly women (n= 19, 61-84?years of age) living in the community were the subjects of this study. [Methods] Blood pressure was measured using an automatic hemodynamometer on the left arm. The calf circumference was measured, and pulse rate was measured on the left radial artery for 1?min by palpation. [Results] No correlations were observed between calf circumference, resting systolic or diastolic pressure, pulse pressure, or mean blood pressure. However, an inverse correlation was observed between calf circumference and resting pulse rate. [Conclusion] Calf circumference measurement may be used as a screening test for resting pulse rate. This test may be useful for the prevention of cardiovascular events.
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Bubble liposomes and ultrasound enhance the antitumor effects of AG73 liposomes encapsulating antitumor agents.
Mol. Pharm.
PUBLISHED: 01-02-2013
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Encapsulating anticancer drugs in liposomes improves their therapeutic window by enhancing antitumor efficacy and reducing side effects. To devise more effective liposomal formulations for antitumor therapy, many research groups have tried to develop tumor-targeting liposomes with enhanced drug release. Previously, we developed doxorubicin (Dox)-encapsulated AG73 peptide-modified liposomes (AG73-Dox), which targeted cancer and endothelial cells, and ultrasound (US) imaging gas-entrapping liposomes, called "Bubble liposomes" (BLs). In this study, to enhance the antitumor effect of AG73-Dox, we combined AG73-Dox with BLs and US. First, to determine whether the addition of BLs and application of US could enhance the cytotoxicity of AG73-Dox, we evaluated the cytotoxicity of the combination of AG73-Dox with BLs and US. BLs and US enhanced cytotoxicity of AG73-Dox more than they enhanced nontargeted Dox-encapsulated liposomes. Next, we examined the intracellular behavior of Dox after treatment with BLs and US. The combination of AG73-Dox with BLs and US did not enhance cellular uptake of Dox, but it did promote drug release in the cytoplasm. To further elucidate the release of Dox in the cytoplasm, we blocked cellular uptake via endosomes at a low temperature. As a result, BLs and US did not have an enhanced drug-release effect until AG73-Dox was taken up into cells. Thus, the combination of AG73-Dox with BLs and US may be useful for cancer therapy as a dual-function drug delivery system with targeted and controlled release.
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Geographical pattern and environmental correlates of regional-scale general flowering in peninsular malaysia.
PLoS ONE
PUBLISHED: 01-01-2013
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In South-East Asian dipterocarp forests, many trees synchronize their reproduction at the community level, but irregularly, in a phenomenon known as general flowering (GF). Several proximate cues have been proposed as triggers for the synchronization of Southeast Asian GF, but the debate continues, as many studies have not considered geographical variation in climate and flora. We hypothesized that the spatial pattern of GF forests is explained by previously proposed climatic cues if there are common cues for GF among regions. During the study, GF episodes occurred every year, but the spatial occurrence varied considerably from just a few forests to the whole of Peninsular Malaysia. In 2001, 2002 and 2005, minor and major GF occurred widely throughout Peninsular Malaysia (GF2001, GF2002, and GF2005), and the geographical patterns of GF varied between the episodes. In the three regional-scale GF episodes, most major events occurred in regions where prolonged drought (PD) had been recorded prior, and significant associations between GF scores and PD were found in GF2001 and GF2002. However, the frequency of PD was higher than that of GF throughout the peninsula. In contrast, low temperature (LT) was observed during the study period only before GF2002 and GF2005, but there was no clear spatial relationship between GF and LT in the regional-scale episodes. There was also no evidence that last GF condition influenced the magnitude of GF. Thus, our results suggest that PD would be essential to trigger regional-scale GF in the peninsula, but also that PD does not fully explain the spatial and temporal patterns of GF. The coarse relationships between GF and the proposed climatic cues may be due to the geographical variation in proximate cues for GF, and the climatic and floristic geographical variations should be considered to understand the proximate factors of GF.
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Important role of the angiotensin II pathway in producing matrix metalloproteinase-9 in human thoracic aortic aneurysms.
J. Surg. Res.
PUBLISHED: 01-01-2013
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The precise pathologic mechanisms underlying human thoracic aortic aneurysms (TAAs) remain uncertain, except that matrix metalloproteinase-9 (MMP-9) is considered a key enzyme for the degradation of extracellular matrix in aneurysm walls. The aim of this study was to elucidate the significance of the angiotensin II (AngII) pathway to MMP-9 production in human TAA walls.
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Novel mechanism for Fc{epsilon}RI-mediated signal transducer and activator of transcription 5 (STAT5) tyrosine phosphorylation and the selective influence of STAT5B over mast cell cytokine production.
J. Biol. Chem.
PUBLISHED: 11-30-2011
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Previous studies indicate that STAT5 expression is required for mast cell development, survival, and IgE-mediated function. STAT5 tyrosine phosphorylation is swiftly and transiently induced by activation of the high affinity IgE receptor, Fc?RI. However, the mechanism for this mode of activation remains unknown. In this study we observed that STAT5 co-localizes with Fc?RI in antigen-stimulated mast cells. This localization was supported by cholesterol depletion of membranes, which ablated STAT5 tyrosine phosphorylation. Through the use of various pharmacological inhibitors and murine knock-out models, we found that IgE-mediated STAT5 activation is dependent upon Fyn kinase, independent of Syk, PI3K, Akt, Brutons tyrosine kinase, and JAK2, and enhanced in the context of Lyn kinase deficiency. STAT5 immunoprecipitation revealed that unphosphorylated protein preassociates with Fyn and that this association diminishes significantly during mast cell activation. SHP-1 tyrosine phosphatase deficiency modestly enhanced STAT5 phosphorylation. This effect was more apparent in the absence of Gab2, a scaffolding protein that docks with multiple negative regulators, including SHP-1, SHP-2, and Lyn. Targeting of STAT5A or B with specific siRNA pools revealed that IgE-mediated mast cell cytokine production is selectively dependent upon the STAT5B isoform. Altogether, these data implicate Fyn as the major positive mediator of STAT5 after Fc?RI engagement and demonstrate importantly distinct roles for STAT5A and STAT5B in mast cell function.
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Bubble liposomes and ultrasound promoted endosomal escape of TAT-PEG liposomes as gene delivery carriers.
Mol. Pharm.
PUBLISHED: 11-04-2011
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We have previously developed laminin-derived AG73 peptide-labeled poly(ethylene glycol)-modified liposomes (AG73-PEG liposomes) for selective cancer gene therapy and reported that Bubble liposomes (BLs) and ultrasound (US) exposure could accelerate the endosomal escape of AG73-PEG liposomes, leading to the enhancement of transfection efficiency; however, it is still unclear whether BLs and US exposure can also enhance the transfection efficiency of other vectors. We therefore assessed the effect of BLs and US exposure on the gene transfection efficiency of trans-activating transcriptor (TAT) peptide modified PEG liposomes. Although TAT-PEG liposomes were efficiently internalized into cells, the efficacy of endosomal escape was insufficient. The transfection efficiencies of TAT-PEG liposomes were enhanced by about 30-fold when BLs and US exposure were used. We also confirmed that BLs and US exposure could not enhance the direct transportation of TAT-PEG liposomes into cells. Confocal microscopy showed that BLs and US exposure promoted endosomal escape of TAT-PEG liposomes. Our results suggested that BLs and US exposure could enhance transfection efficiency by promoting endosomal escape, which was independent of modified molecules of carriers. Thus, BLs and US exposure can be a useful tool to achieve efficient gene transfection by improving endosomal escape of various carriers.
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IRS-2 deficiency in macrophages promotes their accumulation in the vascular wall.
Biochem. Biophys. Res. Commun.
PUBLISHED: 10-04-2011
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The aim of this study was to investigate the role of insulin receptor substrate-2 (IRS-2) mediated signal in macrophages on the accumulation of macrophages in the vascular wall. Mice transplanted with IRS-2(-/-) bone marrow, a model of myeloid cell restricted defect of IRS-2, showed accumulation of monocyte chemoattractant protein-1-expressing macrophages in the vascular wall. Experiments using cultured peritoneal macrophages showed that IRS-2-mediated signal pathway stimulated by physiological concentrations of insulin, not by IL-4, contributed to the suppression of monocyte chemoattractant protein-1 expression induced by lipopolysaccharide. Our data indicated that IRS-2 deficiency in macrophages enhanced their accumulation in the vascular wall accompanied by increased expression of proinflammatory mediators in macrophages. These results suggest a role for insulin resistance in macrophages in early atherosclerogenesis.
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The effects of mechanical stress on the growth, differentiation, and paracrine factor production of cardiac stem cells.
PLoS ONE
PUBLISHED: 09-29-2011
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Stem cell therapies have been clinically employed to repair the injured heart, and cardiac stem cells are thought to be one of the most potent stem cell candidates. The beating heart is characterized by dynamic mechanical stresses, which may have a significant impact on stem cell therapy. The purpose of this study is to investigate how mechanical stress affects the growth and differentiation of cardiac stem cells and their release of paracrine factors. In this study, human cardiac stem cells were seeded in a silicon chamber and mechanical stress was then induced by cyclic stretch stimulation (60 cycles/min with 120% elongation). Cells grown in non-stretched silicon chambers were used as controls. Our result revealed that mechanical stretching significantly reduced the total number of surviving cells, decreased Ki-67-positive cells, and increased TUNEL-positive cells in the stretched group 24 hrs after stretching, as compared to the control group. Interestingly, mechanical stretching significantly increased the release of the inflammatory cytokines IL-6 and IL-1? as well as the angiogenic growth factors VEGF and bFGF from the cells in 12 hrs. Furthermore, mechanical stretching significantly reduced the percentage of c-kit-positive stem cells, but increased the expressions of cardiac troponin-I and smooth muscle actin in cells 3 days after stretching. Using a traditional stretching model, we demonstrated that mechanical stress suppressed the growth and proliferation of cardiac stem cells, enhanced their release of inflammatory cytokines and angiogenic factors, and improved their myogenic differentiation. The development of this in vitro approach may help elucidate the complex mechanisms of stem cell therapy for heart failure.
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Structure-activity relationship studies of S1P agonists with a dihydronaphthalene scaffold.
Bioorg. Med. Chem. Lett.
PUBLISHED: 09-21-2011
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Structure-activity relationship (SAR) of sphingosine-1-phosphate receptor agonists with a dihydronaphthalene scaffold was investigated. Compound 1 was modified to improve S1P(1) agonistic activity and in vivo peripheral lymphocyte lowering (PLL) activity without impairing selectivity over S1P(3) agonistic activity. A detailed SAR study of the terminal lipophilic part revealed that the introduction of substituents on the propylene linker and the terminal benzene ring influences in vitro and PLL activities. Compound 6n bearing a (S)-methyl group at the 2-position on the propylene linker and chlorine at the para-position on the terminal benzene ring showed potent hS1P(1) agonistic activity with excellent selectivity over hS1P(3) and in vivo PLL activity in mice.
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Gene delivery system involving Bubble liposomes and ultrasound for the efficient in vivo delivery of genes into mouse tongue tissue.
Int J Pharm
PUBLISHED: 08-16-2011
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Oral squamous cell carcinoma is the most common type of head and neck cancer. Recently, efficient, easy, and minimally invasive gene delivery methods are expected to be developed as cancer gene therapies. However, the optimal method for delivering therapeutic genes into oral tissue for cancer treatment has not been elucidated. Therefore, we hypothesized that the tongue is a good target tissue for gene delivery with Bubble liposomes and ultrasound. To assess this, we attempted to deliver a mixture of plasmid DNA encoding a luciferase or enhanced green fluorescent protein, and Bubble liposomes into murine tongue with or without ultrasound exposure. The ultrasound conditions were 1 MHz, 2 W/cm(2), 60s, and duty cycle: 50%. The time-course of gene expression in the tongue was investigated with a luciferase assay and fluorescent microscopy. Luciferase expression was significantly increased in tongue transfected using Bubble liposomes and ultrasound compared with that of the tongue untreated with ultrasound, and this high level of luciferase activity was maintained for 2 weeks. From these results, Bubble liposomes can be used in combination with ultrasound to efficiently deliver plasmid DNA into the tongue in vivo. This technique is a highly promising approach for gene delivery into oral tissue.
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Combination of ultrasound and bubble liposome enhance the effect of doxorubicin and inhibit murine osteosarcoma growth.
Cancer Biol. Ther.
PUBLISHED: 08-15-2011
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If ultrasound (US) is applied to cells, permeability across the cell membrane temporarily increases, making it easier for drugs to be taken into the cells from around the cell membrane. Moreover, when used in combination with Bubble liposome (BL: liposomes which entrap an ultrasound imaging gas), even low-power ultrasound can facilitate drug delivery into cells. In the present study, we constructed a new drug delivery system (DDS) involving concomitant use of US and BL with doxorubicin (DOX), a key drug in the chemotherapy of osteosarcoma, and demonstrated both in vitro and in vivo that it markedly inhibited the proliferation of osteosarcoma cells. Furthermore, this system achieved an equivalent antitumor effect at about 1/5 the dose of antitumor agent employed in monotherapy with DOX. These findings suggest the possibility of reduction of adverse events. In this experiment, US and liposomes were tested, both of which are already in use in clinical practice. US and liposomes are both very safe in the body. The DDS composed of these elements we designed can be applied in simple and site-specific fashion and is therefore promising as a new, clinically feasible method of treatment.
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Prophylactic immunization with Bubble liposomes and ultrasound-treated dendritic cells provided a four-fold decrease in the frequency of melanoma lung metastasis.
J Control Release
PUBLISHED: 07-15-2011
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Melanoma has an early tendency to metastasize, and the majority of the resulting deaths are caused by metastatic melanoma. It is therefore important to develop effective therapies for metastasis. Dendritic cell (DC)-based cancer immunotherapy has been proposed as an effective therapeutic strategy for metastasis and recurrence due to prime tumor-specific cytotoxic T lymphocytes. In this therapy, it is important that DCs present peptides derived from tumor-associated antigens on MHC class I molecules. Previously, we developed an innovative approach capable of directly delivering exogenous antigens into the cytosol of DCs using perfluoropropane gas-entrapping liposomes (Bubble liposomes, BLs) and ultrasound. In the present study, we investigated the prevention of melanoma lung metastasis via DC-based immunotherapy. Specifically, antigens were extracted from melanoma cells and used to treat DCs by BL and ultrasound. Delivery into the DCs by this route did not require the endocytic pathway. The delivery efficiency was approximately 74.1%. DCs treated with melanoma-derived antigens were assessed for in vivo efficacy in a mouse model of lung metastasis. Prophylactic immunization with BL/ultrasound-treated DCs provided a four-fold decrease in the frequency of melanoma lung metastases. These in vitro and in vivo results demonstrate that the combination of BLs and ultrasound is a promising method for antigen delivery system into DCs.
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Efficient siRNA delivery using novel siRNA-loaded Bubble liposomes and ultrasound.
Int J Pharm
PUBLISHED: 07-08-2011
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Recently, we developed novel polyethyleneglycol (PEG)-modified liposomes (Bubble liposomes; BLs) entrapping an ultrasound (US) imaging gas and reported that the combination of BLs and US was useful for the delivery of siRNA directly into the cytoplasm. However, the results were obtained using a mixture of BLs and naked siRNA. With systemic injections, it is important to control the biodistribution of both BLs and siRNA. In addition, the delivery of siRNA is affected by nuclease degradation after intravenous administration. In this study, we prepared novel siRNA-loaded BLs (si-BLs) using a cationic lipid, 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP). We demonstrated that siRNA could be loaded onto BLs containing DOTAP and that siRNA-loaded BLs were stable in serum. A specific gene-silencing effect was also achieved by transfection with si-BLs. Thus, the combination of si-BLs with US exposure can be used for delivery of siRNA to a specific tissue via systemic injection.
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A facile preparation method of a PFC-containing nano-sized emulsion for theranostics of solid tumors.
Int J Pharm
PUBLISHED: 06-14-2011
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Theranostics means a therapy conducted in a diagnosis-guided manner. For theranostics of solid tumors by means of ultrasound, we designed a nano-sized emulsion containing perfluoropentane (PFC5). This emulsion can be delivered into tumor tissues through the tumor vasculatures owing to its nano-size, and the emulsion is transformed into a micron-sized bubble upon sonication through phase transition of PFC5. The micron-sized bubbles can more efficiently absorb ultrasonic energy for better diagnostic images and can exhibit more efficient ultrasound-driven therapeutic effects than nano-sized bubbles. For more efficient tumor delivery, smaller size is preferable, yet the preparation of a smaller emulsion is technically more difficult. In this paper, we used a bath-type sonicator to successfully obtain small PFC5-containing emulsions in a diameter of ca. 200nm. Additionally, we prepared these small emulsions at 40°C, which is above the boiling temperature of PFC5. Accordingly, we succeeded in obtaining very small nano-emulsions for theranostics through a very facile method.
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Development of novel nucleic acid-loaded Bubble liposomes using cholesterol-conjugated siRNA.
J Drug Target
PUBLISHED: 06-13-2011
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Recently, we developed polyethyleneglycol (PEG)-modified liposomes (Bubble liposomes; BLs) entrapping ultrasound (US) gas and reported that the combination of BLs and US exposure was an effective tool for the delivery of siRNA directly into cells and US-exposed tissues within a short period; however, the results were obtained using a mixture of BLs and naked siRNA. With systemic injections, it is important to control the biodistribution of both BLs and siRNA. In addition, the delivery of siRNA is affected by nuclease degradation and rapid removal from the circulation after intravenous administration. In this study, we attempted to prepare novel siRNA-loaded BLs (chol-si-BLs) using cholesterol-conjugated siRNA (chol-siRNA). We demonstrated that chol-siRNA could be loaded into BLs, leading to the stability of siRNA even in the presence of an RNase. The specific gene-silencing effect was also achieved by transfection with chol-si-BLs and US. Thus, the combination of chol-si-BLs with US exposure is expected to deliver siRNA into a specific tissue via systemic injection.
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Mast cell interleukin-2 production contributes to suppression of chronic allergic dermatitis.
Immunity
PUBLISHED: 06-08-2011
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The incidence of chronic allergic dermatitis is rapidly increasing. Regulatory control of this disease has not been adequately explored. Here we report that mast cell-derived interleukin-2 (IL-2) contributes to the suppression of chronic allergic dermatitis. Mice deficient in IL-2 production, or deficient in mast cells (Kit(W-sh/W-sh)), showed exacerbated dermatitis upon repeated oxazolone challenge when compared to their wild-type counterparts. Adoptive transfer of wild-type, but not Il2(-/-), mast cells into Kit(W-sh/W-sh) mice dampened the inflammatory response. During the course of disease, mast cell expansion occurred at the site of inflammation and also in the spleen, where production of IL-2 by mast cells was markedly enhanced. In the absence of mast cell IL-2 production, the ratio of activated to regulatory T cells at the site of inflammation was increased. Thus, MC-derived IL-2 contributes to the maintenance of suppression in chronic allergic skin inflammation.
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Technical advance: soluble OX40 molecule mimics regulatory T cell modulatory activity on Fc?RI-dependent mast cell degranulation.
J. Leukoc. Biol.
PUBLISHED: 06-07-2011
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Tregs play a central role in modulating Fc?RI-dependent MC effector functions in the course of the allergic response. Cellular interaction depends on the constitutive expression of OX40 on Tregs and the OX40L counterpart on MCs. Study of OX40L signaling on MCs is hampered by the need of a highly purified molecule, which triggers OX40L specifically. We now report that sOX40 mimics the physiological activity of Treg interaction by binding to activated MCs. When treated with sOX40, activated MCs showed decreased degranulation and Ca(++) influx, whereas PLC-?2 phosphorylation remained unaffected. Once injected into experimental animals, sOX40 not only located within the endothelium but also in parenchyma, where it could be found in close proximity and apparently bound to MCs. This soluble molecule triggers MC-OX40L without the requirement of Tregs, thus allowing study of OX40L signaling pathways in MCs and in other OX40L-expressing cell populations. Importantly, as sOX40 inhibits MC degranulation, it may provide an in vivo therapeutic tool in allergic disease.
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Delayed sternal closure after vacuum-assisted closure therapy for tracheo-innominate artery fistula repair.
Interact Cardiovasc Thorac Surg
PUBLISHED: 05-31-2011
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We report a case of successful innominate artery resection with delayed sternal closure after vacuum-assisted closure (VAC) therapy for a tracheo-innominate artery fistula (TIF). A 42-year-old woman with cerebral palsy underwent tracheostomy for respiratory assistance. On postoperative day 14, she was transferred to our hospital after an episode of massive hemoptysis. TIF was diagnosed based on the findings of multidetector computed tomography. Thus, we resected the innominate artery and started VAC therapy to control the postoperative local infection. The patient recovered uneventfully, without any infectious sequelae. Our strategy, which includes VAC therapy, for TIF repair may eliminate postoperative infective problems that could induce sequential bleeding and sternal compromise. To our knowledge, this is the first report of using VAC therapy for TIF.
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Involvement of activated transcriptional process in efficient gene transfection using unmodified and mannose-modified bubble lipoplexes with ultrasound exposure.
J Control Release
PUBLISHED: 05-06-2011
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Recently, our group developed ultrasound (US)-responsive and mannose-modified gene carriers (Man-PEG(2000) bubble lipoplexes), and successfully obtained a high level of gene expression in mannose receptor-expressing cells following gene transfection using Man-PEG(2000) bubble lipoplexes and US exposure. We also reported that large amounts of plasmid DNA (pDNA) were transferred into the cytoplasm of the targeted cells in the gene transfection using this method. In the present study, we investigated the involvement of transcriptional processes on enhanced gene expression obtained by unmodified and Man-PEG(2000) bubble lipoplexes with US exposure. The transcriptional process related to activator protein-1 (AP-1) and nuclear factor-?B (NF?B) was activated by US exposure, and was founded to be involved in enhanced gene expression obtained by gene transfection using unmodified and Man-PEG(2000) bubble lipoplexes with US exposure. On the other hand, activation of AP-1 and NF?B pathways followed by US exposure was hardly involved in the inflammatory responses in the gene transfection using this method. These findings suggest that activation of AP-1 and NF?B followed by US exposure is involved in the enhanced gene expression using unmodified and Man-PEG(2000) bubble lipoplexes with US exposure, and the selection of pDNAs activated by US exposure is important in this gene transfection method.
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Secondary omental and pectoralis major double flap reconstruction following aggressive sternectomy for deep sternal wound infections after cardiac surgery.
J Cardiothorac Surg
PUBLISHED: 04-18-2011
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Deep sternal wound infection after cardiac surgery carries high morbidity and mortality. Our strategy for deep sternal wound infection is aggressive strenal debridement followed by vacuum-assisted closure (VAC) therapy and omental-muscle flap reconstrucion. We describe this strategy and examine the outcome and long-term quality of life (QOL) it achieves.
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Discovery of S1P agonists with a dihydronaphthalene scaffold.
Bioorg. Med. Chem. Lett.
PUBLISHED: 04-02-2011
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Structure-activity relationship of sphingosine-1-phosphate receptor agonists was examined. Cinnamyl derivative 1 was modified to improve S1P(1) agonistic activity as well as selectivity over S1P(3) agonistic activity. Dihydronaphthalene derivative 10d was identified as a potent S1P(1) receptor agonist with high selectivity against S1P(3) and enhanced efficacy in lowering peripheral lymphocyte counts in mice.
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PKC? regulates hepatic insulin sensitivity and hepatosteatosis in mice and humans.
J. Clin. Invest.
PUBLISHED: 03-30-2011
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C57BL/6J and 129S6/Sv (B6 and 129) mice differ dramatically in their susceptibility to developing diabetes in response to diet- or genetically induced insulin resistance. A major locus contributing to this difference has been mapped to a region on mouse chromosome 14 that contains the gene encoding PKC?. Here, we found that PKC? expression in liver was 2-fold higher in B6 versus 129 mice from birth and was further increased in B6 but not 129 mice in response to a high-fat diet. PRKCD gene expression was also elevated in obese humans and was positively correlated with fasting glucose and circulating triglycerides. Mice with global or liver-specific inactivation of the Prkcd gene displayed increased hepatic insulin signaling and reduced expression of gluconeogenic and lipogenic enzymes. This resulted in increased insulin-induced suppression of hepatic gluconeogenesis, improved glucose tolerance, and reduced hepatosteatosis with aging. Conversely, mice with liver-specific overexpression of PKC? developed hepatic insulin resistance characterized by decreased insulin signaling, enhanced lipogenic gene expression, and hepatosteatosis. Therefore, changes in the expression and regulation of PKC? between strains of mice and in obese humans play an important role in the genetic risk of hepatic insulin resistance, glucose intolerance, and hepatosteatosis; and thus PKC? may be a potential target in the treatment of metabolic syndrome.
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Importin beta plays an essential role in the regulation of the LysRS-Ap(4)A pathway in immunologically activated mast cells.
Mol. Cell. Biol.
PUBLISHED: 03-14-2011
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We recently reported that diadenosine tetraphosphate hydrolase (Ap(4)A hydrolase) plays a critical role in gene expression via regulation of intracellular Ap(4)A levels. This enzyme serves as a component of our newly described lysyl tRNA synthetase (LysRS)-Ap(4)A biochemical pathway that is triggered upon immunological challenge. Here we explored the mechanism of this enzymes translocation into the nucleus and found its immunologically dependent association with importin beta. Silencing of importin beta prevented Ap(4)A hydrolase nuclear translocation and affected the local concentration of Ap(4)A, which led to an increase in microphthalmia transcription factor (MITF) transcriptional activity. Furthermore, immunological activation of mast cells resulted in dephosphorylation of Ap(4)A hydrolase, which changed the hydrolytic activity of the enzyme.
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[Familial aortic dissection of non-Marfan syndrome with mutations in the transforming growth factor-beta receptor type 1 genes].
Kyobu Geka
PUBLISHED: 03-11-2011
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Marfan syndrome is an inherited connective tissue disorder with ocular, skeletal and cardiovascular systems and often causes acute aortic dissection. Interestingly, there have been several reports of familial thoracic aortic dissection in patients with autosomal dominant diseases without Marfan syndrome. Variation of the transforming growth factor-beta receptor (TGFBR) gene is reported to be one of the causes. We report a case of a familial aortic dissection not associated with Marfan syndrome, with mutation of TGFBR type 1. Hereditary aortic dissection of the non-Marfan syndrome that does not have clinical manifestations is not uncommon. Thus, the existence of familial aortic aneurysm should be in mind in diagnosis and treatment.
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The elucidation of gene transferring mechanism by ultrasound-responsive unmodified and mannose-modified lipoplexes.
Biomaterials
PUBLISHED: 03-01-2011
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The development of gene transfection methods enhancing the level of gene expression under simple and low-toxic condition is required for gene therapy in clinical. Our group has developed the ultrasound (US)-mediated gene transfection method using Man-PEG(2000) bubble lipoplexes, which are US-responsive and mannose-modified gene carriers, and succeeded in obtaining the enhanced gene expression in mannose receptor-expressing cells selectively by the gene transfer using Man-PEG(2000) bubble lipoplexes with US exposure in vitro and in vivo. Here, we investigated pDNA transferring mechanism followed by US exposure to unmodified and Man-PEG(2000) bubble lipoplexes, in particular, focused on US exposure timing. Following investigation of intracellular transferring characteristics, a large amount of pDNA was transferred into the cytoplasm followed by US-mediated destruction of bubble lipoplexes in the gene transfer using both bubble lipoplexes with US exposure. Moreover, the effective gene expression was obtained without TNF-? production when US was exposed until 5 min after the addition of bubble lipoplexes. These findings suggest that the gene transfer using unmodified and Man-PEG(2000) bubble lipoplexes with US exposure enables to transfer pDNA into the cytoplasm, and optimized US exposure timing is important to achieve the high level of gene expression and the low level of pro-inflammatory cytokine production.
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Suppression of melanoma growth and metastasis by DNA vaccination using an ultrasound-responsive and mannose-modified gene carrier.
Mol. Pharm.
PUBLISHED: 02-22-2011
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DNA vaccination has attracted much attention as a promising therapy for the prevention of metastasis and relapse of malignant tumors, especially highly metastatic tumors such as melanoma. However, it is difficult to achieve a potent cancer vaccine effect by DNA vaccination, since the number of dendritic cells, which are the major targeted cells of DNA vaccination, is very few. Here, we developed a DNA vaccination for metastatic and relapsed melanoma by ultrasound (US)-responsive and antigen presenting cell (APC)-selective gene carriers reported previously, named Man-PEG???? bubble lipoplexes. Following immunization using US exposure and Man-PEG(2000) bubble lipoplexes constructed with pUb-M, which expresses ubiquitylated melanoma-specific antigens (gp100 and TRP-2), the secretion of Th1 cytokines (IFN-? and TNF-?) and the activities of cytotoxic T lymphocytes (CTLs) were specifically enhanced in the presence of B16BL6 melanoma antigens. Moreover, we succeeded in obtaining potent and sustained DNA vaccine effects against solid and metastatic tumor derived from B16BL6 melanoma specifically. The findings obtained from this study suggest that the gene transfection method using Man-PEG???? bubble lipoplexes and US exposure could be suitable for DNA vaccination aimed at the prevention of metastatic and relapsed cancer.
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Validation of the JapanSCORE versus the logistic EuroSCORE for predicting operative mortality of cardiovascular surgery in Yamaguchi University Hospital.
Gen Thorac Cardiovasc Surg
PUBLISHED: 01-24-2011
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Recent studies have shown that the European System for Cardiac Operative Risk Evaluation (Euro-SCORE) is a reliable risk model in cardiac surgery. In Japan, the JapanSCORE has been devised as the Japanese original risk model for cardiovascular surgery. We evaluated the validity of the JapanSCORE, especially in comparison with the logistic EuroSCORE METHODS: We calculated the predicted mortality according to two risk models for 523 consecutive patients who underwent cardiovascular surgery within a 6-year period (July 2003 to June 2009) at Yamaguchi University Hospital. We assessed the scores validity by calculating the area under the receiver operating characteristics curve (C-index) and by the chi-squared test or Fishers exact test.
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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.

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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.