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Find video protocols related to scientific articles indexed in Pubmed.
[Clinical nutrition in surgery. Guidelines of the German Society for Nutritional Medicine].
Chirurg
PUBLISHED: 04-11-2014
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While enhanced recovery after surgery (ERAS) programs are the standard for perioperative management, special nutritional care has to be administered to malnourished patients and those at metabolic risk with special regard to patients with postoperative complications.
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Voltammetric determination of nitrite in meat products using polyvinylimidazole modified carbon paste electrode.
Food Chem
PUBLISHED: 01-22-2014
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A simple and sensitive voltammetric method was developed to determine the amount of nitrite by using Carbon Paste Electrode (CPE) which is modified with polyvinylimidazole (PVI). A buffer solution of phosphate with a pH 4 value was used in the experiments. The amount of the nitrite-ion was determined by cyclic voltammetry (CV). The electro-chemical behaviour of nitrite-ion was investigated by using CV on the PVI modified CPE. A well-defined oxidation peak was obtained at 0.83 V against a reference Ag/AgCl electrode. Differential pulse voltammetry (DPV) was applied for the calibration plot and for the detection limit. The optimisation procedure was done in two steps: using a two-level factorial design for preliminary evaluation of the contributing factors, and the Box-Behnken Design (BBD) to assess the optimal experimental conditions. These are done with the analysis of 3 different factors in 15 runs of DPV. The optimum conditions are obtained within a linear response range of 5×10(-7)-1×10(-4) mol L(-1). Regression analysis is performed within this range showed the linear equation of y=0.028x+3.93×10(-7) with r(2)=0.9982, and for n=7. Limit of Detection (LOD) was 9×10(-8) mol L(-1) with S/N=3, and Limit of Quantification (LOQ) was 3×10(-7) mol L(-1) with S/N=10. The procedure was used successfully to detect the amount of nitrite in meat products.
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Targeting Sphingosine Kinase Induces Apoptosis and Tumor Regression for KSHV-Associated Primary Effusion Lymphoma.
Mol. Cancer Ther.
PUBLISHED: 10-18-2013
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Sphingosine kinase (SPHK) is overexpressed by a variety of cancers, and its phosphorylation of sphingosine results in accumulation of sphingosine-1-phosphate (S1P) and activation of antiapoptotic signal transduction. Existing data indicate a role for S1P in viral pathogenesis, but roles for SPHK and S1P in virus-associated cancer progression have not been defined. Rare pathologic variants of diffuse large B-cell lymphoma arise preferentially in the setting of HIV infection, including primary effusion lymphoma (PEL), a highly mortal tumor etiologically linked to the Kaposis sarcoma-associated herpesvirus (KSHV). We have found that ABC294640, a novel clinical-grade small molecule selectively targeting SPHK (SPHK2 > SPHK1), induces dose-dependent caspase cleavage and apoptosis for KSHV(+) patient-derived PEL cells, in part through inhibition of constitutive signal transduction associated with PEL cell proliferation and survival. These results were validated with induction of PEL cell apoptosis using SPHK2-specific siRNA, as well as confirmation of drug-induced SPHK inhibition in PEL cells with dose-dependent accumulation of proapoptotic ceramides and reduction of intracellular S1P. Furthermore, we demonstrate that systemic administration of ABC294640 induces tumor regression in an established human PEL xenograft model. Complimentary ex vivo analyses revealed suppression of signal transduction and increased KSHV lytic gene expression within drug-treated tumors, with the latter validated in vitro through demonstration of dose-dependent viral lytic gene expression within PEL cells exposed to ABC294640. Collectively, these results implicate interrelated mechanisms and SPHK2 inhibition in the induction of PEL cell death by ABC294640 and rationalize evaluation of ABC294640 in clinical trials for the treatment of KSHV-associated lymphoma. Mol Cancer Ther; 1-11. ©2013 AACR.
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Sustained PKC?II activity confers oncogenic properties in a phospholipase D- and mTOR-dependent manner.
FASEB J.
PUBLISHED: 10-11-2013
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Protein kinase C (PKC) is a family of serine/threonine kinases implicated in a variety of physiological processes. We have shown previously that sustained activation of the classical PKC? and PKC?II induces their phospholipase D (PLD)-dependent internalization and translocation to a subset of the recycling endosomes defined by the presence of PKC and PLD (the pericentrion), which results in significant differences in phosphorylation of PKC substrates. Here, we have investigated the biological consequences of sustained PKC activity and the involvement of PLD in this process. We find that sustained activation of PKC results in activation of the mammalian target of rapamycin (mTOR)/S6 kinase pathway in a PLD- and endocytosis-dependent manner, with both pharmacologic inhibitors and siRNA implicating the PLD2 isoform. Notably, dysregulated overexpression of PKC?II in A549 lung cancer cells was necessary for the enhanced proliferation and migration of these cancer cells. Inhibition of PKC?II with enzastaurin reduced A549 cell proliferation by >60% (48 h) and migration by >50%. These biological effects also required both PLD activity and mTOR function, with both the PLD inhibitor FIPI and rapamycin reducing cell growth by >50%. Reciprocally, forced overexpression of wild-type PKC?II, but not an F666D mutant that cannot interact with PLD, was sufficient to enhance cell growth and increase migration of noncancerous HEK cells; indeed, both properties were almost doubled when compared to vector control and PKC-F666D-overexpressing cells. Notably, this condition was also dependent on both PLD and mTOR activity. In summary, these data define a PKC-driven oncogenic signaling pathway that requires both PLD and mTOR, and suggest that inhibitors of PLD or mTOR would be beneficial in cancers where PKC overexpression is a contributing or driving factor.-El Osta, M., Liu, M. Adada, M., Senkal, C. E., Idkowiak-Baldys, J., Obeid, L. M., Clarke, C. J., Hannun, Y. A. Sustained PKC?II activity confers oncogenic properties in a phospholipase D- and mTOR-dependent manner.
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Temporal trends of ventilator-associated pneumonia incidence and the effect of implementing health-care bundles in a suburban community.
Chest
PUBLISHED: 08-03-2013
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Recent changes in critical care delivery, including the widespread implementation of health-care bundles, were aimed at reducing complications of critical illness, in particular ventilator-associated pneumonia (VAP), but no population-based study evaluated its effectiveness.
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Metastatic choriocarcinoma: a rare presentation as a neck mass.
Ear Nose Throat J
PUBLISHED: 06-20-2013
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Testicular carcinoma metastatic to the neck is rare. Even more rare is a finding of choriocarcinoma as a neck mass without any sign of a primary testicular tumor, as only a few cases have been reported in the literature. We describe a new case that occurred in a 29-year-old man who presented with a neck mass. Fine-needle aspiration biopsy identified the tumor as a malignant epithelial neoplasm. Radiologic findings indicated the presence of a systemic metastasis of a tumor to the chest and abdomen, as well as the neck. Findings on an incisional biopsy of the neck mass were consistent with a choriocarcinoma. The testicles were normal on palpation and ultrasonography. The patient was diagnosed with metastatic choriocarcinoma with an unknown primary, and he was started on chemotherapy. On the second day of treatment, which was 25 days after his referral to our clinic, he died of respiratory insufficiency.
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Short-term results of Neurelec Digisonic SP cochlear implantation in prelingually deafened children.
Eur Arch Otorhinolaryngol
PUBLISHED: 03-12-2013
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This paper examines the reports on the selection criteria and the post-operative performance of 25 children implanted with the Neurelec Digisonic SP. This study reported benefits from Neurelec Digisonic SP cochlear implant in auditory and speech perception outcomes. There has been a lack of studies into the additional factors such as level of the mothers education and bilingualism, which is a factor that may have a significant effect on the success of cochlear implantation. This paper examines the reports on the reasons for the differences in performance and the post-operative performance of 25 children implanted with the Neurelec Digisonic SP. Meaningful Auditory Integration Scale and Meaningful Use of Speech Scale questionnaires were used just before 3, 6, 12, and 18 months following implantation. Electrode array was inserted without difficulty in all cases, with no complications to date. This is a retrospective and cross-sectional study and all the data were collected between March 2010 and December 2012. Auditory performance improved over time for up to 12 months after implantation. Our experience indicates that the Neurelec Digisonic SP cochlear implant system in children under the age of two is relatively safe and reliable. The Neurelec Digisonic SP device surgery can be performed without complications. Auditory performance results support the effectiveness of early implantation. These important findings further support the importance of professionals working very closely with parents or especially mothers and enhancing their involvement in achieving therapy goals to develop auditory skills and speech in young children following cochlear implantation.
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Effects of voice therapy in school-age children.
J Voice
PUBLISHED: 03-07-2013
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To assess the overall efficacy of voice therapy for dysphonia in school-age children in two different cities in Turkey.
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Effectiveness of the combined hearing and masking devices on the severity and perception of tinnitus: a randomized, controlled, double-blind study.
ORL J. Otorhinolaryngol. Relat. Spec.
PUBLISHED: 01-30-2013
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The aim of this study was to evaluate the effect of combined hearing and tinnitus masking devices that are appropriately programmed for acoustic stimulations using wide-band noise over the specific frequency range of tinnitus.
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Solid phase extraction and determination of nickel in water samples by using novel thiol-containing sulfonamide polymeric resin and atomic absorption spectrophotometer.
Guang Pu Xue Yu Guang Pu Fen Xi
PUBLISHED: 10-20-2011
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Interest in preconcentration techniques for the determination of metals at ultratrace levels still continues increasingly because of some disadvantages of flameless atomic absorption spectrometry as well as the high costs of other sensitive methods in compared to flame atomic absorption spectrometry. In this study, thiol-containing sulfonamide resin was synthesized, characterized and applied as a new sorption material for solid phase extraction of nickel in drinking water samples. After preconcentration procedure, flame atomic absorption spectrometry was used for determinations. Optimum parameters were found to be pH = 3.2, contact time = 20 min and eluate volume = 3 mL. The limit of detection was found to be 0.75 ng x mL(-1). The synthesized resin exhibits the superiority in compared to the other adsorption reagents because of the fact that there is no necessity of any complexing reagent, high sorption capacity as well as the relatively fast extraction rate. The Ni concentrations in the studied 21 kind of water samples were found to be in the range of BDL-4.0 ng x mL(-1).
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Alteration of ceramide synthase 6/C16-ceramide induces activating transcription factor 6-mediated endoplasmic reticulum (ER) stress and apoptosis via perturbation of cellular Ca2+ and ER/Golgi membrane network.
J. Biol. Chem.
PUBLISHED: 10-19-2011
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Mechanisms that regulate endoplasmic reticulum (ER) stress-induced apoptosis in cancer cells remain enigmatic. Recent data suggest that ceramide synthase1-6 (CerS1-6)-generated ceramides, containing different fatty acid chain lengths, might exhibit distinct and opposing functions, such as apoptosis versus survival in a context-dependent manner. Here, we investigated the mechanisms involved in the activation of one of the major ER stress response proteins, ATF-6, and subsequent apoptosis by alterations of CerS6/C(16)-ceramide. Induction of wild type (WT), but not the catalytically inactive mutant CerS6, increased tumor growth in SCID mice, whereas siRNA-mediated knockdown of CerS6 induced ATF-6 activation and apoptosis in multiple human cancer cells. Down-regulation of CerS6/C(16)-ceramide, and not its further metabolism to glucosylceramide or sphingomyelin, activated ATF-6 upon treatment with ER stress inducers tunicamycin or SAHA (suberoylanilide hydroxamic acid). Induction of WT-CerS6 expression, but not its mutant, or ectopic expression of the dominant-negative mutant form of ATF-6 protected cells from apoptosis in response to CerS6 knockdown and tunicamycin or SAHA treatment. Mechanistically, ATF-6 activation was regulated by a concerted two-step process involving the release of Ca(2+) from the ER stores ([Ca(2+)](ER)), which resulted in the fragmentation of Golgi membranes in response to CerS6/C(16)-ceramide alteration. This resulted in the accumulation of pro-ATF-6 in the disrupted ER/Golgi membrane network, where pro-ATF6 is activated. Accordingly, ectopic expression of a Ca(2+) chelator calbindin prevented the Golgi fragmentation, ATF-6 activation, and apoptosis in response to CerS6/C(16)-ceramide down-regulation. Overall, these data suggest a novel mechanism of how CerS6/C(16)-ceramide alteration activates ATF6 and induces ER-stress-mediated apoptosis in squamous cell carcinomas.
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Results of a phase II trial of gemcitabine plus doxorubicin in patients with recurrent head and neck cancers: serum C??-ceramide as a novel biomarker for monitoring response.
Clin. Cancer Res.
PUBLISHED: 07-26-2011
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Here we report a phase II clinical trial, which was designed to test a novel hypothesis that treatment with gemcitabine (GEM)/doxorubicin (DOX) would be efficacious via reconstitution of C(18)-ceramide signaling in head and neck squamous cell carcinoma (HNSCC) patients for whom first-line platinum-based therapy failed.
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Immobilization and stabilization of papain on poly(hydroxyethyl methacrylate-ethylenglycol dimethacrylate) beads grafted with epoxy functional polymer chains via surface-initiated-atom transfer radical polymerization (SI-ATRP).
Bioresour. Technol.
PUBLISHED: 06-01-2011
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Poly(hydroxyethyl methacrylate-ethylen glycol dimethacrylate), p(HEMA-EGDMA), beads were prepared by suspension polymerization, and were decorated with fibrous poly(glycidyl methacrylate), p(GMA), via surface initiated-atom transfer radical polymerization (SI-ATRP). The functional epoxy groups of the beads were used for covalent immobilization of papain. The average amount of immobilized enzyme was 18.7 mg/g beads. The immobilized enzyme was characterized by temperature, pH, operational and storage stability experiments. The maximum velocity of the free and immobilized enzymes (V(max)) and Michaelis-Menten constant (K(m)) values were determined as 10.7 and 8.3 U/mg proteins and 274 and 465 ?M, respectively. The immobilized papain was operated in a batch reactor, and it was very effective for hydrolysis of different proteins (i.e., casein and cytochrom c).
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Concerted functions of HDAC1 and microRNA-574-5p repress alternatively spliced ceramide synthase 1 expression in human cancer cells.
EMBO Mol Med
PUBLISHED: 05-10-2011
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Histone deacetylases (HDACs) and microRNAs (miRs) have pro-survival roles, but the mechanism behind this is unclear. Repression of ceramide synthase 1 (CerS1), altering C(18) -ceramide generation, was linked to drug resistance and metastasis. Here we report that the CerS1 promoter was repressed by HDAC1-dependent inhibition of Sp1 recruitment to two specific GC-boxes spanning the -177 and -139 region. Moreover, an alternatively spliced variant CerS1 mRNA (CerS1-2) was detected mainly in cancer cells or primary tumour tissues compared to controls, which was targeted by miR-574-5p for degradation. A specific 3UTR-targeting site, localized within the retained intron between exons 6 and 7, was identified, and its mutation, or miR-574-5p knockdown prevented the degradation of CerS1-2 mRNA. Interference with HDAC1 and miR-574-5p reconstituted CerS1-2 expression and C(18) -ceramide generation in multiple human cancer cell lines, which subsequently inhibited proliferation and anchorage-independent growth. Accordingly, knockdown of CerS1 partially protected cancer cells from MS-275/miR-574-5p siRNA-mediated growth inhibition. Thus, these data suggest that the HDAC1/miR-574-5p axis might provide a novel therapeutic target to reconstitute tumour suppressor CerS1/ceramide signalling.
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Sphingosine kinase-1 and sphingosine 1-phosphate receptor 2 mediate Bcr-Abl1 stability and drug resistance by modulation of protein phosphatase 2A.
Blood
PUBLISHED: 04-28-2011
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The mechanisms by which sphingosine kinase-1 (SK-1)/sphingosine 1-phosphate (S1P) activation contributes to imatinib resistance in chronic myeloid leukemia (CML) are unknown. We show herein that increased SK-1/S1P enhances Bcr-Abl1 protein stability, through inhibition of its proteasomal degradation in imatinib-resistant K562/IMA-3 and LAMA-4/IMA human CML cells. In fact, Bcr-Abl1 stability was enhanced by ectopic SK-1 expression. Conversely, siRNA-mediated SK-1 knockdown in K562/IMA-3 cells, or its genetic loss in SK-1(-/-) MEFs, significantly reduced Bcr-Abl1 stability. Regulation of Bcr-Abl1 by SK-1/S1P was dependent on S1P receptor 2 (S1P2) signaling, which prevented Bcr-Abl1 dephosphorylation, and degradation via inhibition of PP2A. Molecular or pharmacologic interference with SK-1/S1P2 restored PP2A-dependent Bcr-Abl1 dephosphorylation, and enhanced imatinib- or nilotinib-induced growth inhibition in primary CD34(+) mononuclear cells obtained from chronic phase and blast crisis CML patients, K562/IMA-3 or LAMA4/IMA cells, and 32Dcl3 murine progenitor cells, expressing the wild-type or mutant (Y253H or T315I) Bcr-Abl1 in situ. Accordingly, impaired SK-1/S1P2 signaling enhanced the growth-inhibitory effects of nilotinib against 32D/T315I-Bcr-Abl1-derived mouse allografts. Since SK-1/S1P/S1P2 signaling regulates Bcr-Abl1 stability via modulation of PP2A, inhibition of SK-1/S1P2 axis represents a novel approach to target wild-type- or mutant-Bcr-Abl1 thereby overcoming drug resistance.
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The validity and reliability of the Turkish version of the University of Washington Quality of Life Questionnaire for patients with head and neck cancer.
Am J Otolaryngol
PUBLISHED: 03-23-2011
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The University of Washington Quality of Life Questionnaire (UW-QOL) is an English-language survey used to assess the quality of life of patients with head and neck cancer. The present study aimed to translate this widely used questionnaire into Turkish according to international guidelines and to statistically determine its validity and reliability by administering it to native Turkish-speaking patients.
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Precipitable water modelling using artificial neural network in Çukurova region.
Environ Monit Assess
PUBLISHED: 02-09-2011
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Precipitable water (PW) is an important atmospheric variable for climate system calculation. Local monthly mean PW values were measured by daily radiosonde observations for the time period from 1990 to 2006. Artificial neural network (ANN) method was applied for modeling and prediction of mean precipitable water data in Çukurova region, south of Turkey. We applied Levenberg-Marquardt (LM) learning algorithm and logistic sigmoid transfer function in the network. In order to train our neural network we used data of Adana station, which are assumed to give a general idea about the precipitable water of Çukurova region. Thus, meteorological and geographical data (altitude, temperature, pressure, and humidity) were used in the input layer of the network for Çukurova region. Precipitable water was the output. Correlation coefficient (R(2)) between the predicted and measured values for monthly mean daily sum with LM method values was found to be 94.00% (training), 91.84% (testing), respectively. The findings revealed that the ANN-based prediction technique for estimating PW values is as effective as meteorological radiosonde observations. In addition, the results suggest that ANN method values be used so as to predict the precipitable water.
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Sphingolipids and cancer: ceramide and sphingosine-1-phosphate in the regulation of cell death and drug resistance.
Future Oncol
PUBLISHED: 11-11-2010
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Sphingolipids have emerged as bioeffector molecules, controlling various aspects of cell growth and proliferation in cancer, which is becoming the deadliest disease in the world. These lipid molecules have also been implicated in the mechanism of action of cancer chemotherapeutics. Ceramide, the central molecule of sphingolipid metabolism, generally mediates antiproliferative responses, such as cell growth inhibition, apoptosis induction, senescence modulation, endoplasmic reticulum stress responses and/or autophagy. Interestingly, recent studies suggest de novo-generated ceramides may have distinct and opposing roles in the promotion/suppression of tumors, and that these activities are based on their fatty acid chain lengths, subcellular localization and/or direct downstream targets. For example, in head and neck cancer cells, ceramide synthase 6/C(16)-ceramide addiction was revealed, and this was associated with increased tumor growth, whereas downregulation of its synthesis resulted in ER stress-induced apoptosis. By contrast, ceramide synthase 1-generated C(18)-ceramide has been shown to suppress tumor growth in various cancer models, both in situ and in vivo. In addition, ceramide metabolism to generate sphingosine-1-phosphate (S1P) by sphingosine kinases 1 and 2 mediates, with or without the involvement of G-protein-coupled S1P receptor signaling, prosurvival, angiogenesis, metastasis and/or resistance to drug-induced apoptosis. Importantly, recent findings regarding the mechanisms by which sphingolipid metabolism and signaling regulate tumor growth and progression, such as identifying direct intracellular protein targets of sphingolipids, have been key for the development of new chemotherapeutic strategies. Thus, in this article, we will present conclusions of recent studies that describe opposing roles of de novo-generated ceramides by ceramide synthases and/or S1P in the regulation of cancer pathogenesis, as well as the development of sphingolipid-based cancer therapeutics and drug resistance.
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Is propofol sedation with midazolam induction safe during endoscopic procedures without anesthesiologist?
Hepatogastroenterology
PUBLISHED: 11-02-2010
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Sedation is important for the success and quality of endoscopy. We aimed to evaluate the safety of propofol during the endoscopy under supervision of a gastroenterologist without an anesthesiologist.
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Type I thyroplasty revision 1 year after a window was mistakenly created on the cricoid cartilage.
Ear Nose Throat J
PUBLISHED: 05-13-2010
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Unilateral vocal fold paralysis causes glottic incompetence and can result in significant morbidity. To prevent such morbidity, surgeons treat affected patients with vocal fold medialization techniques; type I thyroplasty medialization surgery is widely used for this purpose. In this procedure, a window is opened on the thyroid cartilage to allow for placement of a silicon prosthesis to medialize the vocal fold. A 38-year-old woman presented to our clinic for evaluation of hoarseness and a low-pitched voice, which we diagnosed as being caused by left vocal fold paralysis. Two years earlier, she had undergone a thyroidectomy for the treatment of benign thyroid disease. One year after that, she underwent type I thyroplasty medialization surgery at another center. During that operation, the surgeon had mistakenly created the window on the cricoid cartilage rather than the thyroid cartilage. When he inserted the silicon prosthesis into the cricoid window, the patient developed acute respiratory obstruction. At that point, the prosthesis was removed and the operation was terminated. One year later, she presented to us, and we performed a revision type I thyroplasty. Intraoperatively, we discovered that the original window had been opened on the cricoid cartilage instead of the thyroid cartilage, which was intact. We left the cricoid window untouched, opened a new window on the thyroid cartilage, and completed the type I thyroplasty in the usual fashion. The patients postoperative recovery was uneventful, and she was doing well at 5 years of follow-up. To the best of our knowledge, no case of a thyroplasty window being opened on a cricoid cartilage has been reported in the literature.
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Removal of dyes from water using crosslinked aminomethane sulfonic acid based resin.
Environ Geochem Health
PUBLISHED: 04-17-2010
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A new polymeric resin with amino sulfonic acid pendant functions has been prepared for the extraction of acidic and basic dyes from water. Beaded polymer supports were prepared by suspension polymerization of vinyl benzyl chloride (0.9 mol) and ethylene glycol dimethacrylate (0.1 mol). The resulting copolymer beads were modified with amino methane sulfonic acid. The dye adsorption capacity of the resin was found as 0.16 g dye/g resin for ramazol black and 0.15 g dye/g resin for crystal violet. The pH depending measurements and dye sorption kinetics of the resin were also investigated.
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Severe odynophagia in a patient developing after azithromycin intake: a case report.
Cases J
PUBLISHED: 02-03-2010
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Drug-induced esophageal ulcers most commonly cause heartburn, midsternal pain and dysphagia. In our clinic azithromycin is a relative widely used antibiotic for respiratory tract infections and otitis media because of its activity against Haemophilus influenzae and atypical pathogens, and its ease of administration. After a thorough search in Pubmed the present case is the first one to report azithromycin-induced esophageal ulcer and associated symptoms in the literature.
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National assessment of sea level rise using topographic and census data for Turkish coastal zone.
Environ Monit Assess
PUBLISHED: 10-23-2009
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Turkish coastal zone elevation to sea level rise was illustrated by using digital elevation model and Geographical information systems methods. It was intended to determine several parameters such as population, settlements, land use, wetlands, contribution to national agricultural production and taxes at risk by using high resolution SRTM topographic, orthorectified Landsat Thematic Mapper Mosaics and census data with GIS methods within 0-10 m elevation of national level. All parameters were examined for coastal cities, coastal districts, settlements and villages status. As a result of the analysis of data set, it was found that approximately 7,319 km(2) of land area lies below 10 m contour line in Turkey, and is hence highly vulnerable to sea-level rise. 28 coastal cities, 191 districts and 181 villages or towns are located below 10 m contour line in study area. In the short term, for the struggle of negative impact of sea level rise, the findings suggest that the Ministry of Environment should declare new areas as protection areas and develop special environmental programs for national level.
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Antiapoptotic roles of ceramide-synthase-6-generated C16-ceramide via selective regulation of the ATF6/CHOP arm of ER-stress-response pathways.
FASEB J.
PUBLISHED: 09-01-2009
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Emerging results suggest that ceramides with different fatty acid chain lengths might play distinct functions in the regulation of tumor growth and therapy. Here we report that de novo-generated C(18)- and C(16)-ceramides by ceramide synthases 1 and 6 (CerS1 and CerS6) play opposing proapoptotic and prosurvival roles, respectively, in human head and neck squamous cell carcinomas (HNSCCs). Unexpectedly, knockdown of CerS6/C(16)-ceramide using small interfering RNA induced endoplasmic reticulum (ER)-stress-mediated apoptosis. Reconstitution of C(16)-ceramide generation by induced expression of wild-type CerS6, but not its catalytically inactive mutant, protected cells from cell death induced by knockdown of CerS6. Moreover, using molecular tools coupled with analysis of sphingolipid metabolism showed that generation of C(16)-ceramide, and not dihydro-C(16)-ceramide, by induced expression of CerS6 rescued cells from ER stress and apoptosis. Mechanistically, regulation of ER-stress-induced apoptosis by CerS6/C(16)-ceramide was linked to the activation of a specific arm, ATF6/CHOP, of the unfolded protein response pathway. Notably, while expression of CerS1/C(18)-ceramide inhibited HNSCC xenograft growth, CerS6/C(16)-ceramide significantly protected ER stress, leading to enhanced tumor development and growth in vivo, consistent with their pro- and antiapoptotic roles, respectively. Thus, these data reveal an unexpected and novel prosurvival role of CerS6/C(16)-ceramide involved in the protection against ER-stress-induced apoptosis and induction of HNSCC tumor growth.
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A new mixed-backbone oligonucleotide against glucosylceramide synthase sensitizes multidrug-resistant tumors to apoptosis.
PLoS ONE
PUBLISHED: 05-12-2009
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Enhanced ceramide glycosylation catalyzed by glucosylceramide synthase (GCS) limits therapeutic efficiencies of antineoplastic agents including doxorubicin in drug-resistant cancer cells. Aimed to determine the role of GCS in tumor response to chemotherapy, a new mixed-backbone oligonucleotide (MBO-asGCS) with higher stability and efficiency has been generated to silence human GCS gene. MBO-asGCS was taken up efficiently in both drug-sensitive and drug-resistant cells, but it selectively suppressed GCS overexpression, and sensitized drug-resistant cells. MBO-asGCS increased doxorubicin sensitivity by 83-fold in human NCI/ADR-RES, and 43-fold in murine EMT6/AR1 breast cancer cells, respectively. In tumor-bearing mice, MBO-asGCS treatment dramatically inhibited the growth of multidrug-resistant NCI/ADR-RE tumors, decreasing tumor volume to 37%, as compared with scrambled control. Furthermore, MBO-asGCS sensitized multidrug-resistant tumors to chemotherapy, increasing doxorubicin efficiency greater than 2-fold. The sensitization effects of MBO-asGCS relied on the decreases of gene expression and enzyme activity of GCS, and on the increases of C(18)-ceramide and of caspase-executed apoptosis. MBO-asGCS was accumulation in tumor xenografts was greater in other tissues, excepting liver and kidneys; but MBO-asGCS did not exert significant toxic effects on liver and kidneys. This study, for the first time in vivo, has demonstrated that GCS is a promising therapeutic target for cancer drug resistance, and MBO-asGCS has the potential to be developed as an antineoplastic agent.
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Direct interaction between the inhibitor 2 and ceramide via sphingolipid-protein binding is involved in the regulation of protein phosphatase 2A activity and signaling.
FASEB J.
PUBLISHED: 04-02-2009
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In this study, the inhibitor 2 of protein phosphatase 2A (I2PP2A) was identified in vitro and in situ as a ceramide-binding protein, which exhibits stereoisomer specificity and fatty acid chain length preference. Site- directed mutagenesis coupled with structural details of I2PP2A suggested that VIK 207-209 residues localized on helix 7 are important for ceramide binding and single mutation of K209D altered this interaction. Notably, I2PP2A-ceramide binding decreased the association between PP2A and the inhibitor, preventing the inhibition of PP2A activity in vitro. In addition, studies in A549 human lung cancer cells revealed that ceramide mediates c-Myc degradation via its PP2A-dependent dephosphorylation at S62, and treatment with okadaic acid and expression of c-Myc mutants with S62A or S62D conversions resulted in resistance to ceramide-mediated degradation. Importantly, whereas down-regulation of I2PP2A enhanced PP2A-mediated c-Myc degradation in response to ceramide, ectopic expression of wild-type I2PP2A but not of its K209D mutant protected this degradation in A549 cells. Moreover, expression of wild-type I2PP2A prevented the growth-inhibitory effects of ceramide both against A549 cells and xenograft-driven tumors in situ and in vivo compared with that in controls. Thus, these results suggest that direct interaction of I2PP2A with ceramide plays important biological roles via the regulation of PP2A activity and signaling, which in turn control ceramide-mediated degradation of c-Myc and antiproliferation.
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Poly(glycidylmethacrylate) brushes generated on poly(VBC) beads by SI-ATRP technique: hydrazine and amino groups functionalized for invertase adsorption and purification.
J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.
PUBLISHED: 03-12-2009
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Crosslinked-poly(vinylbenzylchloride), poly(VBC), beads were prepared by suspension polymerization and poly(glycidylmethacrylate) was grafted by surface-initiated-atom radical polymerization (SI-ATRP) technique. Epoxy groups of the grafted poly(GMA) were reacted with hydrazine and ammonia to create an affinity binding sites. The hydrazine and amine functionalized poly(VBC-g-GMA) beads were used as an affinity support for adsorption of invertase from solution and yeast crude extract. The influence of pH, equilibrium time, ionic strength and initial invertase concentration on the adsorption capacities of both hydrazine and amine functionalized beads has been investigated. Maximum invertase adsorptions onto hydrazine and amine functionalized beads, were 86.7 and 30.4 mg/g at pH 4.0 and 5.5, respectively. The experimental equilibrium data fitted well to the Temkin isotherm model. Finally, the hydrazine functionalized poly(VBC-g-GMA) beads were used for the purification of invertase from crude yeast extract in a batch system and the purity of the eluted invertase from the hydrazine functionalized beads was determined as 92% by HPLC from single step purification protocol.
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Lipid emulsions - Guidelines on Parenteral Nutrition, Chapter 6.
Ger Med Sci
PUBLISHED: 01-14-2009
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The infusion of lipid emulsions allows a high energy supply, facilitates the prevention of high glucose infusion rates and is indispensable for the supply with essential fatty acids. The administration of lipid emulsions is recommended within < or =7 days after starting PN (parenteral nutrition) to avoid deficiency of essential fatty acids. Low-fat PN with a high glucose intake increases the risk of hyperglycaemia. In parenterally fed patients with a tendency to hyperglycaemia, an increase in the lipid-glucose ratio should be considered. In critically ill patients the glucose infusion should not exceed 50% of energy intake. The use of lipid emulsions with a low phospholipid/triglyceride ratio is recommended and should be provided with the usual PN to prevent depletion of essential fatty acids, lower the risk of hyperglycaemia, and prevent hepatic steatosis. Biologically active vitamin E (alpha-tocopherol) should continuously be administered along with lipid emulsions to reduce lipid peroxidation. Parenteral lipids should provide about 25-40% of the parenteral non-protein energy supply. In certain situations (i.e. critically ill, respiratory insufficiency) a lipid intake of up to 50 or 60% of non-protein energy may be reasonable. The recommended daily dose for parenteral lipids in adults is 0.7-1.3 g triglycerides/kg body weight. Serum triglyceride concentrations should be monitored regularly with dosage reduction at levels >400 mg/dl (>4.6 mmol/l) and interruption of lipid infusion at levels >1000 mg/dl (>11.4 mmol/l). There is little evidence at this time that the choice of different available lipid emulsions affects clinical endpoints.
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Determination of lead in milk and yoghurt samples by solid phase extraction using a novel aminothioazole-polymeric resin.
Food Chem
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A preconcentration method was developed by using a new aminothioazole-containing sulfonamide resin in solid phase extraction for the determination of lead and nickel in milk and yoghurt samples. The optimisation of experimental conditions was performed. The optimum parameters for Pb and Ni were found to be 3.5, 30 min and 2.5 mL for pH, contact time, eluate volume, respectively. After preconcentration step, atomic absorption spectrometry was used for the determinations. The enhancement of 350- and 50-fold in the sensitivity of Pb and Ni were achieved by combination of the slotted tube atom trap-atomic absorption spectrometry with the optimised preconcentration method, respectively. Limits of detection were found to be 0.15 ng mL?¹ for Pb and 0.75 ng mL?¹ for Ni. The lead concentrations in the studied samples were found to be in the range of 15-61 ng Pb mL?¹ for milk and 21-42 ng Pb g?¹ for yoghurt samples.
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Sphingosine analogue drug FTY720 targets I2PP2A/SET and mediates lung tumour suppression via activation of PP2A-RIPK1-dependent necroptosis.
EMBO Mol Med
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Mechanisms that alter protein phosphatase 2A (PP2A)-dependent lung tumour suppression via the I2PP2A/SET oncoprotein are unknown. We show here that the tumour suppressor ceramide binds I2PP2A/SET selectively in the nucleus and including its K209 and Y122 residues as determined by molecular modelling/simulations and site-directed mutagenesis. Because I2PP2A/SET was found overexpressed, whereas ceramide was downregulated in lung tumours, a sphingolipid analogue drug, FTY720, was identified to mimick ceramide for binding and targeting I2PP2A/SET, leading to PP2A reactivation, lung cancer cell death, and tumour suppression in vivo. Accordingly, while molecular targeting of I2PP2A/SET by stable knockdown prevented further tumour suppression by FTY720, reconstitution of WT-I2PP2A/SET expression restored this process. Mechanistically, targeting I2PP2A/SET by FTY720 mediated PP2A/RIPK1-dependent programmed necrosis (necroptosis), but not by apoptosis. The RIPK1 inhibitor necrostatin and knockdown or genetic loss of RIPK1 prevented growth inhibition by FTY720. Expression of WT- or death-domain-deleted (DDD)-RIPK1, but not the kinase-domain-deleted (KDD)-RIPK1, restored FTY720-mediated necroptosis in RIPK1(-/-) MEFs. Thus, these data suggest that targeting I2PP2A/SET by FTY720 suppresses lung tumour growth, at least in part, via PP2A activation and necroptosis mediated by the kinase domain of RIPK1.
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Ceramide targets autophagosomes to mitochondria and induces lethal mitophagy.
Nat. Chem. Biol.
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Mechanisms by which autophagy promotes cell survival or death are unclear. We provide evidence that C(18)-pyridinium ceramide treatment or endogenous C(18)-ceramide generation by ceramide synthase 1 (CerS1) expression mediates autophagic cell death, independent of apoptosis in human cancer cells. C(18)-ceramide-induced lethal autophagy was regulated via microtubule-associated protein 1 light chain 3 ?-lipidation, forming LC3B-II, and selective targeting of mitochondria by LC3B-II-containing autophagolysosomes (mitophagy) through direct interaction between ceramide and LC3B-II upon Drp1-dependent mitochondrial fission, leading to inhibition of mitochondrial function and oxygen consumption. Accordingly, expression of mutant LC3B with impaired ceramide binding, as predicted by molecular modeling, prevented CerS1-mediated mitochondrial targeting, recovering oxygen consumption. Moreover, knockdown of CerS1 abrogated sodium selenite-induced mitophagy, and stable LC3B knockdown protected against CerS1- and C(18)-ceramide-dependent mitophagy and blocked tumor suppression in vivo. Thus, these data suggest a new receptor function of ceramide for anchoring LC3B-II autophagolysosomes to mitochondrial membranes, defining a key mechanism for the induction of lethal mitophagy.
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Manganese-superoxide dismutase and glutathione peroxidase 1 polymorphisms in recurrent tonsillitis and tonsillar hypertrophy.
Int. J. Pediatr. Otorhinolaryngol.
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To investigate the association of manganese-superoxide dismutase and glutathione peroxidase 1 polymorphisms with susceptibility to recurrent tonsillitis and tonsillar hypertrophy.
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Comparison of early postoperative pain among surgical techniques for obstructive sleep apnea.
Eur Arch Otorhinolaryngol
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One of the criticized aspects of surgeries for obstructive sleep apnea (OSA) is postoperative pain. We performed a study to compare the severity of pain occurring after different surgical techniques and to determine analgesic requirements in the first postoperative 24 h. Forty-eight patients with primary snoring or OSA who underwent anterior palatoplasty (AP), lateral pharyngoplasty (LP) or tongue base suspension suture (TBS) were included in this study. A visual analog scale (VAS) was used for measuring pain intensity. Tramadol with patient-controlled analgesia (PCA) device and when necessary rescue pethidine was used for pain relief. VAS pain scores, total PCA-tramadol consumptions and requirement of rescue analgesic in AP, LP and TBS groups were compared. Pain scores in TBS group were higher than AP group in all of the study time points except at 12th hour and LP group until the 10th hour. When compared with AP group, VAS was significantly higher in LP group at the 1st hour. Mean total tramadol consumptions were significantly different between the groups (AP-LP, p = 0.039; AP-TBS, p < 0.001; LP-TBS, p < 0.001). It was highest in the TBS group and lowest in the AP group. In the LP group, three patients (16.7 %) needed rescue analgesia in comparison with 11 (73.3 %) in the TBS group. None of the patients in the AP group needed rescue analgesic. AP is the least painful and TBS is the most painful procedure. PCA-bolus tramadol effectively treats pain caused by AP and LP; however, alleviation of pain caused by TBS usually needs rescue opioid analgesic.
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Relationship between procalcitonin levels and presence of vesicoureteral reflux during first febrile urinary tract infection in children.
Urology
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To investigate the association between the procalcitonin (PCT) level during the first febrile urinary tract infection (UTI) in children and the presence of vesicoureteral reflux (VUR). VUR-associated UTI is among the primary causes of chronic renal failure in Turkey.
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Therapeutical potential of autologous peripheral blood mononuclear cell transplantation in patients with type 2 diabetic critical limb ischemia.
J. Diabetes Complicat.
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The aim was to evaluate the therapeutic effectiveness of granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood mononuclear cells (PBMNCs) in critical limb ischemia (CLI) of type 2 diabetic patients.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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