JoVE Visualize What is visualize?
Stop Reading. Start Watching.
Advanced Search
Stop Reading. Start Watching.
Regular Search
Find video protocols related to scientific articles indexed in Pubmed.
GPER-1 and Estrogen Receptor-? Ligands Modulate Aldosterone Synthesis.
Endocrinology
PUBLISHED: 08-28-2014
Show Abstract
Hide Abstract
Fertile women have lower blood pressure and cardiovascular risk than age-matched men, which suggests that estrogens exert cardiovascular protective effects. However, whether 17 ?-estradiol (E2) blunts aldosterone secretion, and thereby affects the gender dimorphism of blood pressure, is unknown. We therefore sought for the estrogen receptor (ER) subtypes in human adrenocortical tissues ex vivo by performing gene and protein expression studies. We also investigated the effect of E2 on aldosterone synthesis and the involved receptors through in vitro functional experiments in the adrenocortical cells HAC15. We found that in the human adrenal cortex and aldosterone-producing adenoma cells, the most expressed ERs were the ER? and the G protein-coupled receptor-1 (GPER-1), respectively. After selective ER? blockade, E2 (10 nmol/L) markedly increased both the expression of aldosterone synthase and the production of aldosterone (+5- to 7-fold vs baseline, P < .001). Under the same condition, the GPER-1 receptor agonist 1-[4-(6-bromo-benzo (1, 3)dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c] quinolin-8-yl]-ethanone (G-1) (10 nmol/L) mimicked this effect, which was abrogated by cotreatment with either the GPER-1 receptor antagonist (3aS*,4R*,9bR*)-4-(6-Bro-mo-1,3-benzodioxol-5-yl)-3a,4,5,9b-3H-cyclopenta[c]quinoline (G-15), or a selective protein kinase A inhibitor 8-Bromo-2-monobutyryladenosine-3,5-cyclic mono-phosphorothioate, Rp-isomer. Silencing of the ER? significantly raised aldosterone synthase expression and aldosterone production. Conversely, silencing of the GPER-1 lowered aldosterone synthase gene and protein expression. Moreover, it blunted the stimulatory effect of E2 on aldosterone synthase that was seen during ER? blockade. These results support the conclusion that in humans, E2 inhibits aldosterone synthesis by acting via ER?. Pharmacologic disinhibition of ER? unmasks a potent secretagogue effect of E2 that involves GPER-1 and protein kinase A signaling.
Related JoVE Video
Programmed cell death 4 and microRNA 21 inverse expression is maintained in cells and exosomes from ovarian serous carcinoma effusions.
Cancer Cytopathol
PUBLISHED: 04-16-2014
Show Abstract
Hide Abstract
Ovarian serous carcinoma (OSC) is a fatal gynecologic malignancy usually presenting with bilateral localization and malignant peritoneal effusion. Programmed cell death 4 (PDCD4) is a tumor suppressor gene whose expression is directly controlled by microRNA-21 (miR-21). Exosomes are small cell-derived vesicles that participate in intercellular communication, delivering their cargo of molecules to specific cells. Exosomes are involved in several physiological and pathological processes including oncogenesis, immunomodulation, angiogenesis, and metastasis. The current study analyzed the expression of PDCD4 and miR-21 in resected OSC specimens and in cells and exosomes from OSC peritoneal effusions.
Related JoVE Video
Inflammation and pancreatic cancer: molecular and functional interactions between S100A8, S100A9, NT-S100A8 and TGF?1.
Cell Commun. Signal
PUBLISHED: 03-08-2014
Show Abstract
Hide Abstract
In order to gain further insight on the crosstalk between pancreatic cancer (PDAC) and stromal cells, we investigated interactions occurring between TGF?1 and the inflammatory proteins S100A8, S100A9 and NT-S100A8, a PDAC-associated S100A8 derived peptide, in cell signaling, intracellular calcium (Cai2+) and epithelial to mesenchymal transition (EMT). NF-?B, Akt and mTOR pathways, Cai2+ and EMT were studied in well (Capan1 and BxPC3) and poorly differentiated (Panc1 and MiaPaCa2) cell lines.
Related JoVE Video
YAP/TAZ incorporation in the ?-catenin destruction complex orchestrates the Wnt response.
Cell
PUBLISHED: 03-05-2014
Show Abstract
Hide Abstract
The Hippo transducers YAP/TAZ have been shown to play positive, as well as negative, roles in Wnt signaling, but the underlying mechanisms remain unclear. Here, we provide biochemical, functional, and genetic evidence that YAP and TAZ are integral components of the ?-catenin destruction complex that serves as cytoplasmic sink for YAP/TAZ. In Wnt-ON cells, YAP/TAZ are physically dislodged from the destruction complex, allowing their nuclear accumulation and activation of Wnt/YAP/TAZ-dependent biological effects. YAP/TAZ are required for intestinal crypt overgrowth induced by APC deficiency and for crypt regeneration ex vivo. In Wnt-OFF cells, YAP/TAZ are essential for ?-TrCP recruitment to the complex and ?-catenin inactivation. In Wnt-ON cells, release of YAP/TAZ from the complex is instrumental for Wnt/?-catenin signaling. In line, the ?-catenin-dependent maintenance of ES cells in an undifferentiated state is sustained by loss of YAP/TAZ. This work reveals an unprecedented signaling framework relevant for organ size control, regeneration, and tumor suppression.
Related JoVE Video
Multiple sporadic gastrointestinal stromal tumors concomitant with ampullary adenocarcinoma: a case report with KIT and PDGFRA mutational analysis and miR-221/222 expression profile.
Pathol. Res. Pract.
PUBLISHED: 01-08-2014
Show Abstract
Hide Abstract
GISTs originating multifocally at different GI sites, in patients lacking familial syndromes, could be interpreted as recurrent/metastatic disease. MiR-221/222 have recently been identified as regulators of KIT expression in GISTs. We report the first case of synchronous GISTs in the stomach and duodenum concomitant with an ampullary adenocarcinoma. Different CD117 expression patterns could be related to different KIT mutational status in the two lesions: gastric GIST showed a dot-like pattern and lacked KIT mutations; duodenal GIST had a strong membranous expression pattern, likely due to KIT exon 9 duplication, which is associated with lower response to imatinib. MiR-221/222 were downregulated in GISTs as compared with normal tissue (p<0.05) and expressed increased levels in the gastric GIST as compared with duodenal one (p<0.05). Our data support an independent origin of the two GISTs. Determining whether these tumors are multiple primaries or recurrencies is helpful to predict their malignancy and to select proper treatment.
Related JoVE Video
LowER EXPRESSION OF THE TWIK-RELATED ACID-SENSITIVE K+ CHANNEL 2 (TASK-2) GENE IS A HALLMARK OF ALDOSTERONE PRODUCING ADENOMA CAUSING HUMAN PRIMARY ALDOSTERONISM.
J. Clin. Endocrinol. Metab.
PUBLISHED: 11-27-2013
Show Abstract
Hide Abstract
Context.The molecular mechanisms of primary aldosteronism, a common cause of human hypertension, are unknown, but alterations of K(+) channels can play a key role.Objective.To investigate: i) the expression of the Twik-related Acid-Sensitive K(+) channels (TASK) in Aldosterone Producing Adenomas (APAs); ii) the role of TASK-2 in aldosterone synthesis; iii) the determinants of TASK-2 blunted expression in APA.Design.We analyzed the transcriptome and the microRNA profiles of 32 consecutive APA and investigated the protein expression and localization of TASK-2 in APA and adrenocortical cell lines (H295R and HAC15) using immunoblotting and confocal microscopy. The functional effect of TASK-2 blunted activity caused by a dominant negative mutation on steroidogenic enzymes and aldosterone production was also assessed. TASK-2 regulation by selected microRNA was studied by a luciferase assay.Results.TASK-2 was consistently less expressed at the transcript and protein level in APAs than in the normal human adrenal cortex. H295R cells transfection with a TASK-2 dominant negative mutant construct significantly increased the aldosterone production by 153% and the gene expression of Aldosterone synthase (CYP11B2, gene expression fold change 3.1 vs control, p<0.05) and of the Steroidogenic acute regulatory protein (STAR) (gene expression fold change 1.8 vs control, p <0.05). Two microRNAs - hsa-miR-23 and hsa-miR-34- were found to decrease TASK-2 expression by binding to the 3 UTR of the TASK-2 gene.Conclusions.The TASK-2 channel lower expression represents a hallmark of APA and is associated to a higher expression of hsa-miR-23 and hsa-miR-34. The ensuing blunted TASK-2 activity increased the production of aldosterone in vitro and the expression of STAR and CYP11B2. Hence, the lower expression of TASK-2 channel in APA cells can explain high aldosterone secretion in human primary aldosteronism in spite of the suppression of angiotensin II, the hypertension and the hypokalemia.
Related JoVE Video
The p53-p66Shc apoptotic pathway is dispensable for tumor suppression whereas the p66Shc-generated oxidative stress initiates tumorigenesis.
Curr. Pharm. Des.
PUBLISHED: 10-01-2013
Show Abstract
Hide Abstract
Reactive oxygen species (ROS) are regarded as hazardous by-products of mitochondrial respiration. In addition to the respiratory chain, specific ROS-generating systems have evolved. In particular, p66Shc is a mitochondrial redox protein that oxidizes cytochrome c to generate H2O2. Consistently, the deletion of p66Shc in cells and tissue results in reduced levels of ROS and oxidative stress. Taking advantage of the p66Shc knock out (p66KO) mouse model of decreased ROS production, we assessed the role of endogenously-produced ROS in tumorigenesis. Spontaneous tumor incidence was investigated and found unaltered in two different strains, 129Sv and C57Bl/6J, p66KO mice. In addition, papilloma formation upon exposure to ultraviolet radiation (UV) or 7,12-Dimethylbenz(a)anthracene/12-O-tetradecanoylphorbol- 13-acetate (DMBA/TPA) was found to be slightly lower in the absence of p66Shc. The role of p66Shc in tumorigenesis was also investigated in the absence of the tumor suppressor gene p53 (p53KO) by generating p53-p66Shc double knock out (DKO) mice. Notably, DKO mice displayed a significantly increased lifespan compared to p53KO mice. In addition, 2-deoxy-2-(18F)fluoro-D-glucose Positron Emission Tomography ([18F]FDG PET) analysis allowed to determine that disease onset occurred later in life in DKO mice compared to p53KO and that a low percentage of these mice did not develop tumors. Overall, these results indicate that although tumor incidence is not decreased in p66KO mice, p66Shc contributes to tumor initiation, in particular upon activation by carcinogens as well as when p53- mediated tumor suppression mechanisms defect.
Related JoVE Video
A 4-MicroRNA signature can discriminate primary lymphomas from anaplastic carcinomas in thyroid cytology smears.
Cancer Cytopathol
PUBLISHED: 08-06-2013
Show Abstract
Hide Abstract
Anaplastic thyroid carcinoma (ATC) and primary thyroid lymphoma (PTL) are uncommon tumors of the thyroid gland with several overlapping clinical and pathologic features that may render their differentiation difficult in fine-needle aspiration (FNA) cytology. MicroRNA (miRNA) signatures have been recently reported as useful diagnostic tools applied to cytology specimens.
Related JoVE Video
1?,25-Dihydroxyvitamin D3 inhibits the human H295R cell proliferation by cell cycle arrest: A model for a protective role of vitamin D receptor against adrenocortical cancer.
J. Steroid Biochem. Mol. Biol.
PUBLISHED: 07-22-2013
Show Abstract
Hide Abstract
Using the human H295R adrenocortical carcinoma cell line as a model, we analyzed the role of 1?,25-dihydroxyvitamin D3 [1?,25(OH)2D3)]-vitamin D receptor (VDR) axis in the growth of adrenocortical cancer (ACC). The presence of VDR in various adrenocortical tissues, including ACC, was also investigated. DNA synthesis was evaluated by [(3)H]thymidine cell incorporation after treatment with 1?,25(OH)2D3 at increasing doses. The effect of 1?,25(OH)2D3 on cell cycle and apoptosis was analyzed with a flow cytometer. Cyclin-dependent kinase 4 (CDK4) expression, a molecular marker of G1-S cell cycle transition phase, was evaluated in cells treated with 1?,25(OH)2D3 before and after VDR gene silencing. 1?,25(OH)2D3 treatment inhibited cell proliferation by 20% at a dose of 1nM, in parallel with steroid secretion decrease. A cell cycle arrest in G1, with no change in apoptotic cell proportion, was observed after 10nM 1?,25(OH)2D3 cell exposure. CDK4 activation was reduced by 10nM 1?,25(OH)2D3 but was not affected by 1?,25(OH)2D3 after VDR gene silencing. Expression of VDR mRNA was lower in ACC than in benign adrenocortical tumors. VDR immunostaining was evident in benign tumors but it was weak in ACC tissues.
Related JoVE Video
The reticulin algorithm for adrenocortical tumor diagnosis: a multicentric validation study on 245 unpublished cases.
Am. J. Surg. Pathol.
PUBLISHED: 06-19-2013
Show Abstract
Hide Abstract
The pathologic diagnosis of adrenocortical carcinoma (ACC) still needs to be improved, because the renowned Weiss Score (WS) system has a poor reproducibility of some parameters and is difficult to apply in borderline cases and in ACC variants. The "reticulin algorithm" (RA) defines malignancy through an altered reticulin framework associated with 1 of the 3 following parameter: necrosis, high mitotic rate, and vascular invasion. This study aimed at validating the interobserver reproducibility of reticulin stain evaluation in an unpublished series of 245 adrenocortical tumors (61 adenomas and 184 carcinomas) from 5 Italian centers, classified according to the WS. Eight pathologists reviewed all reticulin-stained slides. After training, a second round of evaluation on discordant cases was performed 10 weeks later. The RA reclassified 67 cases (27%) as adenomas, including 44 with no reticulin alterations and 23 with an altered reticulin framework but lacking the subsequent parameters of the triad. The other 178 cases (73%) were carcinomas according to the above-mentioned criteria. A complete (8/8 pathologists) interobserver agreement was reached in 75% of cases (?=0.702), irrespective of case derivation, pathologists experience, and histologic variants, and was further improved when only those cases with high WS and clinically malignant behavior were considered. After the training, the overall agreement increased to 86%. We conclude that reticulin staining is a reliable technique and an easy-to-interpret system in adrenocortical tumors; moreover, it has a high interobserver reproducibility, which supports the notion of using such a method in the proposed 2-step RA approach for ACC diagnosis.
Related JoVE Video
Predicting late recurrence in surgically treated patients with Cushings disease.
Clin. Endocrinol. (Oxf)
PUBLISHED: 05-06-2013
Show Abstract
Hide Abstract
Cushings disease (CD) has an uncertain prognosis because patients achieving remission after transsphenoidal pituitary neurosurgery (TSS) may relapse. We aimed to identify factors predicting relapse, focusing on desmopressin (DDAVP) and corticotropin-releasing hormone (CRH) tests after surgery.
Related JoVE Video
Munchausen syndrome: a novel cause of drug-resistant hypertension.
J. Hypertens.
PUBLISHED: 04-26-2013
Show Abstract
Hide Abstract
A young patient presented with a history of resistant arterial hypertension, associated with disabling symptoms. He was subjected to an enormous number of tests to identify a pheochromocytoma that was never found. He was eventually discovered to make factitious use of amphetamine to mimic this condition in order to gain medical attention. Munchausen syndrome was thus diagnosed. The patient was discharged and was lost to follow-up until he presented again in 2012 for resistant hypertension in our outpatient clinic. He reported that because of poor blood pressure, he had been referred to a Cardiology department where transcatheter renal denervation was performed with no effect on blood pressure. Thereafter, he was presented to an Endocrinology unit where a left adrenalectomy was performed with diagnosis of pheochromocytoma that was not found at pathology.
Related JoVE Video
Human amniotic fluid stem cells protect rat lungs exposed to moderate hyperoxia.
Pediatr. Pulmonol.
PUBLISHED: 02-17-2013
Show Abstract
Hide Abstract
Treatment of bronchopulmonary dysplasia (BPD) remains as yet an unmet clinical need and recently stem cells have been proposed as a therapeutic tool in animal models. We investigated the role of amniotic fluid stem cells (AFS) in an adult rat model of hyperoxia lung injury.
Related JoVE Video
Detection of microRNAs in archival cytology urine smears.
PLoS ONE
PUBLISHED: 01-22-2013
Show Abstract
Hide Abstract
MicroRNAs dysregulation and profiling have been demonstrated to be clinically relevant in urothelial carcinoma (UC). Urine cytology is commonly used as the mainstay non-invasive test for secondary prevention and follow-up of UC patients. Ancillary tools are needed to support cytopathologists in the diagnosis of low-grade UC. The feasibility and reliability of microRNAs profiling by qRT-PCR analysis (miR-145 and miR-205) in archival routine urine cytology smears (affected by fixation/staining [Papanicolau] and room temperature storage) was tested in a series of 15 non-neoplastic and 10 UC urine specimens. Only samples with >5,000 urothelial cells and with <50% of inflammatory cells/red blood cells clusters were considered. Overall, a satisfactory amount of total RNA was obtained from all the considered samples (mean 1.27±1.43 µg, range 0.06-4.60 µg). Twenty nanograms of total RNA have been calculated to be the minimal total RNA concentration for reliable and reproducible miRNAs expression profiling analysis of archival cytological smears (slope= -3.4084; R-squared=0.99; efficiency=1.94). miR-145 and miR-205 were significantly downregulated in UC samples in comparison to non-tumor controls. These findings demonstrate that urine archival cytology smears are suitable for obtaining high-quality RNA to be used in microRNAs expression profiling. Further studies should investigate if miRNAs profiling can be successfully translated into clinical practice as diagnostic or prognostic markers.
Related JoVE Video
Next-Generation Sequencing of Lung Cancer EGFR Exons 18-21 Allows Effective Molecular Diagnosis of Small Routine Samples (Cytology and Biopsy).
PLoS ONE
PUBLISHED: 01-01-2013
Show Abstract
Hide Abstract
Selection of lung cancer patients for therapy with tyrosine kinase inhibitors directed at EGFR requires the identification of specific EGFR mutations. In most patients with advanced, inoperable lung carcinoma limited tumor samples often represent the only material available for both histologic typing and molecular analysis. We defined a next generation sequencing protocol targeted to EGFR exons 18-21 suitable for the routine diagnosis of such clinical samples. The protocol was validated in an unselected series of 80 small biopsies (n=14) and cytology (n=66) specimens representative of the material ordinarily submitted for diagnostic evaluation to three referral medical centers in Italy. Specimens were systematically evaluated for tumor cell number and proportion relative to non-neoplastic cells. They were analyzed in batches of 100-150 amplicons per run, reaching an analytical sensitivity of 1% and obtaining an adequate number of reads, to cover all exons on all samples analyzed. Next generation sequencing was compared with Sanger sequencing. The latter identified 15 EGFR mutations in 14/80 cases (17.5%) but did not detected mutations when the proportion of neoplastic cells was below 40%. Next generation sequencing identified 31 EGFR mutations in 24/80 cases (30.0%). Mutations were detected with a proportion of neoplastic cells as low as 5%. All mutations identified by the Sanger method were confirmed. In 6 cases next generation sequencing identified exon 19 deletions or the L858R mutation not seen after Sanger sequencing, allowing the patient to be treated with tyrosine kinase inhibitors. In one additional case the R831H mutation associated with treatment resistance was identified in an EGFR wild type tumor after Sanger sequencing. Next generation sequencing is robust, cost-effective and greatly improves the detection of EGFR mutations. Its use should be promoted for the clinical diagnosis of mutations in specimens with unfavorable tumor cell content.
Related JoVE Video
Molecular typing of lung adenocarcinoma on cytological samples using a multigene next generation sequencing panel.
PLoS ONE
PUBLISHED: 01-01-2013
Show Abstract
Hide Abstract
Identification of driver mutations in lung adenocarcinoma has led to development of targeted agents that are already approved for clinical use or are in clinical trials. Therefore, the number of biomarkers that will be needed to assess is expected to rapidly increase. This calls for the implementation of methods probing the mutational status of multiple genes for inoperable cases, for which limited cytological or bioptic material is available. Cytology specimens from 38 lung adenocarcinomas were subjected to the simultaneous assessment of 504 mutational hotspots of 22 lung cancer-associated genes using 10 nanograms of DNA and Ion Torrent PGM next-generation sequencing. Thirty-six cases were successfully sequenced (95%). In 24/36 cases (67%) at least one mutated gene was observed, including EGFR, KRAS, PIK3CA, BRAF, TP53, PTEN, MET, SMAD4, FGFR3, STK11, MAP2K1. EGFR and KRAS mutations, respectively found in 6/36 (16%) and 10/36 (28%) cases, were mutually exclusive. Nine samples (25%) showed concurrent alterations in different genes. The next-generation sequencing test used is superior to current standard methodologies, as it interrogates multiple genes and requires limited amounts of DNA. Its applicability to routine cytology samples might allow a significant increase in the fraction of lung cancer patients eligible for personalized therapy.
Related JoVE Video
Epithelial-mesenchymal transition in malignant mesothelioma.
Mod. Pathol.
PUBLISHED: 10-07-2011
Show Abstract
Hide Abstract
Epithelial-mesenchymal transition is a physiopathological process by which epithelial cells acquire mesenchymal shape and properties. Malignant mesothelioma is histologically characterized by the concomitant presence of epithelioid and sarcomatoid features, the latter being associated to worse prognosis, thus suggesting a role of epithelial-mesenchymal transition in this dual phenotype. We studied 109 malignant mesotheliomas (58 epithelioid, 26 sarcomatoid, and 25 biphasic) by immunohistochemistry and qRT-PCR analysis, and demonstrated a substantial switch from epithelial markers (E-cadherin, ?-catenin, and cytokeratins 5/6) to mesenchymal markers (N-cadherin, vimentin, ?-smooth muscle actin, Snail, Slug, Twist, ZEB1, ZEB2, S100A4, MMP2, and MMP9) through epithelioid to biphasic and sarcomatoid histotypes. In agreement with these findings, the ectopic expression of miR-205 (a repressor of ZEB1 and ZEB2 expression) in MeT-5A (mesothelial cell line), H2452 (an epithelioid malignant mesothelioma cell line) and MSTO-211H (a biphasic malignant mesothelioma cell line) not only induced a significant reduction of ZEB1 and ZEB2 and a consequent up-regulation of E-cadherin gene expression, but also inhibited migration and invasion. Moreover, miR-205 was significantly down-regulated in biphasic and sarcomatoid histotypes (qRT-PCR and in situ hybridization analyses). Collectively, our findings indicate that epithelial-mesenchymal transition has a significant part in the morphological features of malignant mesothelioma. In particular, miR-205 down-regulation correlated significantly with both a mesenchymal phenotype and a more aggressive behavior.
Related JoVE Video
Concurrent primary aldosteronism and subclinical cortisol hypersecretion: a prospective study.
J. Hypertens.
PUBLISHED: 07-02-2011
Show Abstract
Hide Abstract
Primary aldosteronism is the most frequent cause of secondary hypertension and is responsible for an increased risk of cardiometabolic complications. A concomitant subtle cortisol hyperproduction could enhance cardiovascular risk. We prospectively estimated the occurrence of subclinical hypercortisolism in primary aldosteronism patients.
Related JoVE Video
Classification of non-small cell lung carcinoma in transthoracic needle specimens using microRNA expression profiling.
Chest
PUBLISHED: 05-26-2011
Show Abstract
Hide Abstract
Emerging targeted lung cancer therapies require the accurate morphologic subclassification of non-small cell lung cancer (NSCLC), even in scant and distorted specimens obtained by transthoracic needle aspiration (TTNA). MicroRNAs (miRNAs) are small noncoding genes recently reported as useful in differentiating squamous cell carcinoma (SCC) from adenocarcinoma (AD) in resected tumor specimens. We investigated their ability to do so in TTNA specimens.
Related JoVE Video
Axillary node sampling in conjunction with sentinel node biopsy in patients with breast cancer. A prospective preliminary study.
Anticancer Res.
PUBLISHED: 03-08-2011
Show Abstract
Hide Abstract
In patients with breast cancer (BC), axillary lymph node sampling (ALNS) is a reliable procedure with low morbidity, alternative or complementary to sentinel lymph node biopsy (SLNB), which may improve the detection rate of axillary node metastases as compared to SLNB alone in staging the axilla. The aim of this study was to assess the usefulness of ALNS in conjunction with SLNB in improving the sensitivity of SLNB alone at frozen section examination. One hundred and twelve women (median age 56 years, range 29-71 years) with BC underwent SLNB using a combined radioisotope and isosulfan blue dye technique. Two groups of age- and tumor size-matched patients were prospectively randomized: Group A (SLNB alone, 55 women) and group B (SLNB plus ALNS, 57 women). Intraoperative examination showed SN involvement in 32 (28.6%) patients: group A = 14 (25.5%), group B = 18 (31.6%), whilst the final pathology showed axillary node involvement in 7 further cases (group A = 5, group B = 2). The sensitivity and accuracy were 73.7% vs. 90.0% (p = 0.23) and 90.9% vs. 94.7% (p = 0.49), group A vs. B, respectively. Multivariate analysis showed that age >65 years and body mass index independently correlated with the amount of axillary drainage in both groups, which was 47.5 ± 11.3 and 49.6 ± 12.2 ml (A vs. B, p = NS), respectively. In conclusion, in our preliminary study, ALNS in conjunction with SLNB is a low-risk procedure, useful to reduce the false-negative rate of SLNB and to improve the accuracy of intraoperative evaluation of the axillary nodes in patients with BC.
Related JoVE Video
Is genetic screening indicated in apparently sporadic pheochromocytomas and paragangliomas?
Surgery
PUBLISHED: 02-26-2011
Show Abstract
Hide Abstract
Pheochromocytoma (Pheo) is usually considered a sporadic disease. Recently, an increasing rate of genetically based tumors has been reported. However, the need for systematic screening of unsuspected germline mutations in apparently sporadic forms is still debated. This study aimed to assess the effective rate of germline mutations causing Pheo and Paraganglioma (PGL), and the role of systematic genetic screening.
Related JoVE Video
Fine-needle cytology of cutaneous juvenile xanthogranuloma and langerhans cell histiocytosis.
Cancer Cytopathol
PUBLISHED: 09-29-2010
Show Abstract
Hide Abstract
In pediatric patients, a cutaneous nodule is usually diagnosed by performing an excisional biopsy, but fine-needle cytology (FNC) is a safer and noninvasive diagnostic method widely used to obtain diagnostic specimens with little stress to the patient. The authors compared the ability of FNC and biopsy to differentiate Langerhans cell histiocytosis (LCH) from juvenile xanthogranuloma (JXG).
Related JoVE Video
Fine-needle aspiration cytology and (99m)Tc-pertechnetate scintigraphy together in patients with differentiated thyroid carcinoma.
Anticancer Res.
PUBLISHED: 08-05-2010
Show Abstract
Hide Abstract
The aim of this study was to evaluate the usefulness of fine-needle aspiration cytology (FNAC) and (99m)Tc-pertechnetate scintigraphy (TS) together in patients with differentiated thyroid carcinoma. Data from a series of 357 patients (284 women and 73 men, median age 43 years, range 19-73) with solitary thyroid nodule and no signs of hyperfunction, who had undergone both FNAC and TS prior to surgery, were retrospectively reviewed. FNAC distinguished 3 groups of TN (benign, follicular neoplasm, cancer), while patients with cold TN were considered at risk of having a thyroid tumor. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were: 95%, 21%, 20%, 95% and 34% for TS; 82%, 99%, 96%, 96% and 96% for FNAC; 98%, 99%, 97%, 98%, and 99% for TS and FNAC together, respectively. In conclusion, patients with cold TN and FNAC suggesting follicular neoplasm should be considered at risk of having cancer.
Related JoVE Video
A case of anaplastic thyroid cancer with long-term survival.
Anticancer Res.
PUBLISHED: 06-10-2010
Show Abstract
Hide Abstract
Anaplastic thyroid carcinoma (ATC) (less than 10% of all thyroid cancer) is a high-grade neoplasm, characterized by an aggressive clinical course and refractoriness to currently available local and systemic modalities of treatment. It is considered the most aggressive solid tumour, there is no adequate therapy for this disease and few patients with ATC live more than 1 year following diagnosis. We report herein an unusual case of ATC in a 59-year-old woman. She presented to our Institute in December 2004. She received many kinds of chemotherapeutical and multimodal treatment; we obtained a long period of localized disease (about two years) and an excellent response to therapy. She is still alive 58 months from diagnosis.
Related JoVE Video
Differential diagnosis of lung nodules: breast cancer metastases and lung tuberculosis.
Infez Med
PUBLISHED: 04-29-2010
Show Abstract
Hide Abstract
In a follow-up a 74-year-old woman with breast cancer (clinical stage T4N1M0 at onset, treatment by surgical resection and tamoxifen) presented a combination of two distinct diseases in the lung: breast cancer metastasis and tuberculosis. A CT scan showed multiple pulmonary nodular lesions and in only one lesion fine needle aspiration cytology (FNAC) diagnosed tuberculosis. After specific antibiotic therapy, isoniazide and rifampin, a CT scan highlighted disappearance of tubercular lesion. Because occurrence of tuberculosis during chemo or hormone therapy for metastatic breast cancer is rare, the present case is noteworthy. Indeed, it is worth pointing out the differential diagnosis of pulmonary nodular lesions in patients with cancer and the possible reactivation of tuberculosis even in patients without specific symptoms, without typical tubercular radiological features.
Related JoVE Video
A kidney tumor in an adolescent with severe hypertension and hypokalemia: an uncommon case--case report and review of the literature on reninoma.
Urol. Int.
PUBLISHED: 03-08-2010
Show Abstract
Hide Abstract
This report presents a case of a 16-year-old hypertensive boy who presented to our clinic. Laboratory findings showed severe hypokalemia and markedly increased plasma renin activity. Abdominal ultrasonography and contrast-enhanced computed tomography of the abdomen revealed a well-circumscribed, solid, hypoenhancing cortical lesion (2 cm) in the lower pole of the left kidney. The patient underwent nephron-sparing surgery. Histopathologic examination gave a diagnosis of juxtaglomerular cell tumor. Reninoma is an uncommon cause of hypertension in a young adult and should be included in the differential diagnosis as a potential life-threatening and curable condition. The conservative surgical management is the gold standard for small, circumscribed lesions.
Related JoVE Video
Isolation of human adrenocortical aldosterone-producing cells by a novel immunomagnetic beads method.
Endocrinology
PUBLISHED: 01-22-2010
Show Abstract
Hide Abstract
We detected intense CD56 immunostaining in the zona glomerulosa (ZG) and medulla of the normal human adrenal gland and therefore identified CD56, the neural cell adhesion molecule, as a membrane antigen specific for the ZG, aldosterone-producing adenoma (APA), and chromaffin cells. The APA and pheochromocytoma cells, which are histogenetically derived from the ZG and medulla, respectively, also showed intense CD56 immunostaining. Based on these findings we developed a strategy for isolating cells from the ZG and APA using CD56 immunobinding to magnetic beads. Morphology, gene expression studies, and aldosterone measurement confirmed that CD56 positive (+) cells were ZG and APA cells. Analysis of CD56+ cells under light and phase contrast microscopy evidenced that these cells formed clumps, as the ZG cells usually do; with electron microscopy they showed multiple features typical of a steroidogenic phenotype. Expression levels of the CD56 and the aldosterone synthase (CYP11B2) gene were markedly higher in CD56+ cells than CD56- cells (+1600 and +2100% increase, respectively). Moreover, aldosterone secretion was higher (+1380%) from CD56+ cells than from CD56- cells. Hence, this novel methodology allows isolation of a pure population of ZG and APA cells exhibiting multiple characteristics of the aldosterone-producing cells.
Related JoVE Video
FNA cytology and frozen section examination in patients with follicular lesions of the thyroid gland.
Anticancer Res.
PUBLISHED: 12-31-2009
Show Abstract
Hide Abstract
Patients with solitary thyroid nodules should have fine-needle aspiration (FNA) cytology as the initial screening test, but the most of those referred to a surgeon usually undergo frozen section examination (FS). The aim of this retrospective study was to assess the usefulness of FNA cytology and FS together in patients with a solitary thyroid nodule (TN). Two-hundred and ten patients with a TN and FNA cytology suggesting follicular neoplasm underwent intraoperative FS and subsequent hemithyroidectomy or total thyroidectomy. There were 47 (22.4%) men and 163 (77.6%) women, with a median age of 43 years (range 18-76 years). In all patients, ultrasound-guided FNA was successfully performed using 22-G needle prior to surgery. Smears of the FNA samples were stained by May-Grünwald-Giemsa stain and evaluated immediately by the cytologist. Final histology was follicular carcinoma in 23 (10.9%), follicular adenoma in 181 (86.2%), and hyperplasia in 6 (2.9%) patients. No difference (p=NS) in age of the patients, and greatest diameter on the TN was found between groups. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 13.0%, 97.3%, 37.5%, 90.0%, and 88.1% for FNA cytology, and 17.4%, 100%, 100%, 90.8%, and 91.0% for FS, respectively. The combination of FNA plus FS did not significantly improve the results. In conclusion, both FNA cytology and FS are highly specific tests, but their sensitivity is low, even when they are used in combination. Thus, in patients with smears suggesting follicular neoplasm, FS should be considered unnecessary because it does not affect the intraoperative decision making. FS is most useful in those cases that are diagnosed as suspicious for papillary carcinoma by FNA.
Related JoVE Video
P66Shc signals to age.
Aging (Albany NY)
PUBLISHED: 05-07-2009
Show Abstract
Hide Abstract
Oxygen metabolism is thought to impact on aging through the formation of reactive oxygen species (ROS) that are supposed to damage biological molecules. The study of p66(Shc), a crucial regulator of ROS level involved in aging dysfunction, suggests that the incidence of degenerative disease and longevity are determined by a specific signaling function of ROS other than their unspecific damaging property.
Related JoVE Video
Usefulness of combined sestamibi scintimammography, axillary ultrasonography and FNA cytology in reducing the number of sentinel node procedures in patients with early-stage breast cancer.
Anticancer Res.
PUBLISHED: 04-01-2009
Show Abstract
Hide Abstract
Intraoperative analysis of the sentinel lymph node (SLN) status is currently performed in patients with breast cancer (BC) undergoing surgery. Axillary node (AN) metastases are present in up to 60% of cases, but the risk is only 30% in patients with early stage (T1) BC. The aim of this study was to evaluate the usefulness of 99mTc-sestamibi scintimammography (SSM), axillary ultrasonography (US) and US-guided fine-needle aspiration (FNA) cytology together in detecting axillary metastases preoperatively and their potential role in reducing the number of SLN procedures.
Related JoVE Video
Expression and functional role of urotensin-II and its receptor in the adrenal cortex and medulla: novel insights for the pathophysiology of primary aldosteronism.
J. Clin. Endocrinol. Metab.
PUBLISHED: 03-31-2009
Show Abstract
Hide Abstract
The involvement of urotensin II, a vasoactive peptide acting via the G protein-coupled urotensin II receptor, in arterial hypertension remains contentious.
Related JoVE Video
Detection of silica particles in lung tissue by environmental scanning electron microscopy.
Inhal Toxicol
PUBLISHED: 03-24-2009
Show Abstract
Hide Abstract
For pathologists, pneumologists, and occupational and environmental physicians it is relevant to know silica levels in lung tissue to better define limits of exposure. Environmental Scanning Electron Microscopy (ESEM) has been employed to detect silica particles and to compare silica levels in subjects with and without Lung Cancer (LC). We investigated 25 paraffin-embedded tissue samples of patients with LC adenocarcinoma, and 20 fresh samples of subjects without LC deceased for extra-pulmonary diseases. Silica levels were quantified considering the Number of Spots of silica particles (NS), and the Number of Positive Zones (NPZ) in which there was at least one spot. Levels of NS and NPZ were assessed with Poisson-type regression models, and in two samples of silica-exposed workers with LC the performance of models were evaluated. LC patients displayed higher silica levels, as compared to controls; smoking, age and gender had no significant effects on this relationship. Values of NS and NPZ for the exposed workers were in agreement with model estimates. The fitted model between NS and NPZ might be useful in evaluating new observations and in the development of threshold limit values of silica in biological tissues. ESEM is a rapid, simple and valid tool for the determination of silica levels in lung tissues.
Related JoVE Video
Survivin expression impacts prognostically on NSCLC but not SCLC.
Lung Cancer
Show Abstract
Hide Abstract
Survivin is expressed in lung cancer and in most cancer tissues and has a significant impact on prognosis. This work aimed to comparatively assess survivin expression and significance in Non-Small (NSCLC) and Small Cell Lung Cancers (SCLC). Sixty-five NSCLC and 35 SCLC samples were analyzed by semi-quantitative real-time RT-PCR. Survivin mRNA levels were significantly higher in tumors than in normal tissue, and in SCLC than in NSCLC samples. Immunohistochemistry and FISH analyses were performed in 59 and 26 tumor specimens, respectively. In SCLC survivin was only present in cytoplasm, while in some NSCLC cases it also showed nuclear or mixed patterns. FISH analysis did not disclose survivin gene amplification, except for one NSCLC case. Finally, 90 samples were genotyped for the -31G/C SNP of survivin promoter by direct sequencing; the -31G/C SNP genotype status showed a significant association only with nodal NSCLC metastasis, but not with survivin expression in any tumor group. A better prognosis was correlated to higher levels of survivin mRNA and to the presence of at least one G allele at -31 SNP in NSCLC, while these parameters did not correlate with overall survival in SCLC. Moreover, this SNP would appear to have no effect on the risk of lung cancer in our samples. The different prognostic role played by survivin in NSCLC and SCLC highlights the biological differences between these lung tumor histotypes and stresses the need to clarify the molecular pathways leading to their neoplastic transformation.
Related JoVE Video
Unilateral adrenal hyperplasia: a novel cause of surgically correctable primary hyperaldosteronism.
Surgery
Show Abstract
Hide Abstract
Primary hyperaldosteronism may be caused by an aldosterone-producing adenoma (APA), which is correctable by unilateral adrenalectomy or by idiopathic adrenal hyperplasia, a bilateral disease without any indication for surgery. This study sought to assess the prevalence and the results of surgery in unilateral adrenal hyperplasia (UAH).
Related JoVE Video
The miR-17-92 microRNA cluster: a novel diagnostic tool in large B-cell malignancies.
Lab. Invest.
Show Abstract
Hide Abstract
Diffuse large B-cell lymphoma (DLBCL) can present as de novo or can arise through the transformation of many indolent lymphomas, including follicular lymphoma (FL). The morphological differentiation between germinal center-DLBCL (GC-DLBCL) and high-grade (grade 3) FL could be challenging; the accurate sub-classification of large B-cell lymphomas is mandatory in order to select the most appropriate among the new-targeted therapies. Recent expression profiling studies reported microRNAs (miRNAs) (and miR-17-92 cluster, in particular) as useful tools in differentiating DLBCL and FL. However, these preliminary results are based on cell line-derived data or did not consider grade 3 FL cases. To investigate this point, 36 cases of GC-DLBCL and 18 cases of grade 3 non-transforming FL were considered. All diagnoses were based on the World Health Organization criteria and were confirmed by clinical, histological, and immunohistochemical data. Six members of the miR-17-92 cluster (ie, miR-18b, miR-19b, miR-20a, miR-92, miR-93, and miR-106a) and two control miRNAs (ie, miR-150 and miR-210) were quantified by quantitative reverse transcription-polymerase chain reaction. All the considered miR-17-92 cluster miRNAs were significantly overexpressed in GC-DLBCL, being miR-20a and miR-106a the most dysregulated (P<0.001). Receiver operating characteristics (ROCs) analysis was used to find the optimal cut-off in distinguishing the two histotypes. The ROC estimated thresholds for miR-18b, miR-19b, miR-20a, miR-92, and miR-106a displayed a sensitivity level higher than 0.80 in achieving the GC-DLBCL diagnosis. The classification tree built on the six thresholds allowed the correct identification of 35/36 GC-DLBCL (97.2%). Profiling the miR-17-92 cluster is a promising investigative method for differentiating GC-DLBCL from high-grade FL. Subject to the validation of these findings in further larger studies; miR-17-92 cluster could represent a reliable, standardizable diagnostic tool for the sub-classification of large B-cell lymphoid neoplasm.
Related JoVE Video
Hyperparathyroidism can be useful in the identification of primary aldosteronism due to aldosterone-producing adenoma.
Hypertension
Show Abstract
Hide Abstract
Hyperparathyroidism represents as a novel feature of primary aldosteronism (PA). Its occurrence in patients with the surgically correctable aldosterone-producing adenoma (APA) and not in those with bilateral adrenal hyperplasia suggested that the measurement of parathyroid hormone could help in differentiating between these subtypes of PA. To test this hypothesis we measured the plasma levels of intact parathyroid hormone, Ca(2+), and several markers of calcium/phosphorus metabolism in 132 hypertensive patients, including 74 with primary (essential) hypertension and 58 consecutive PA patients. Of the latter, 46 were conclusively diagnosed as APA (by finding of lateralized aldosterone excess, pathology, correction of the hyperaldosteronism, and evidence of a fall of blood pressure after adrenalectomy) and 12 as bilateral adrenal hyperplasia. Based on these diagnoses we used the area under the receiver operator characteristic curve analysis to assess the accuracy of serum parathyroid hormone for identifying the PA cases in the whole group and for distinguishing between APA and bilateral adrenal hyperplasia. In this selected population of hypertensive patients for identifying PA cases, the accuracy of serum parathyroid hormone tended to be lower than that of the aldosterone:renin ratio. However, for discriminating between APA and bilateral adrenal hyperplasia patients it was better than that under the identity line and also that for the aldosterone:renin ratio for pinpointing APA cases among patients with PA. Hence, these findings indicate that raised serum parathyroid hormone levels are a feature of APA that can be useful for selecting the PA patients to be submitted to adrenal vein sampling.
Related JoVE Video
Combination of sorafenib and everolimus impacts therapeutically on adrenocortical tumor models.
Endocr. Relat. Cancer
Show Abstract
Hide Abstract
Treatment options are insufficient in patients with adrenocortical carcinoma (ACC). Based on the efficacy of sorafenib, a tyrosine kinase inhibitor, and everolimus, an inhibitor of the mammalian target of rapamycin in tumors of different histotype, we aimed at testing these drugs in adrenocortical cancer models. The expression of vascular endothelial growth factor and its receptors (VEGFR1-2) was studied in 18 ACCs, 33 aldosterone-producing adenomas, 12 cortisol-producing adenomas, and six normal adrenal cortex by real-time PCR and immunohistochemistry and by immunoblotting in SW13 and H295R cancer cell lines. The effects of sorafenib and everolimus, alone or in combination, were tested on primary adrenocortical cultures and SW13 and H295R cells by evaluating cell viability and apoptosis in vitro and tumor growth inhibition of tumor cell line xenografts in immunodeficient mice in vivo. VEGF and VEGFR1-2 were detected in all samples and appeared over-expressed in two-thirds of ACC specimens. Dose-dependent inhibition of cell viability was observed particularly in SW13 cells after 24?h treatment with either drug; drug combination produced markedly synergistic growth inhibition. Increasing apoptosis was observed in tumor cells treated with the drugs, particularly with sorafenib. Finally, a significant mass reduction and increased survival were observed in SW13 xenograft model undergoing treatment with the drugs in combination. Our data suggest that an autocrine VEGF loop may exist within ACC. Furthermore, a combination of molecularly targeted agents may have both antiangiogenic and direct antitumor effects and thus could represent a new therapeutic tool for the treatment of ACC.
Related JoVE Video
L-citrulline prevents alveolar and vascular derangement in a rat model of moderate hyperoxia-induced lung injury.
Lung
Show Abstract
Hide Abstract
Moderate normobaric hyperoxia causes alveolar and vascular lung derangement in the newborn rat. Endogenous nitric oxide (NO), which promotes lung growth, is produced from the metabolism of L-arginine to L-citrulline in endothelial cells. We investigated whether administering L-citrulline by raising the serum levels of L-arginine and enhancing NO endogenous synthesis attenuates moderate hyperoxia-induced lung injury.
Related JoVE Video
Myxoinflammatory fibroblastic sarcoma: report of a case and review of the literature.
Pediatr. Dev. Pathol.
Show Abstract
Hide Abstract
Myxoinflammatory fibroblastic sarcoma (MIFS) is a low-grade sarcoma generally arising in adults. We present a case of MIFS in a 5-year-old boy with a palpable nodule in the subcutaneous tissue of the scalp. We carried out a literature review to evaluate the diagnostic patterns based on histologic and cytologic features and possible pitfalls and misdiagnoses. A systematic search for articles of interest published between 1995 and 2011 was performed in MEDLINE and PubMed using the words "myxoinflammatory fibroblastic sarcoma," "myxohyaline tumor," and "inflammatory myxoid tumor." Histology and cytology have a pivotal role in the differential diagnosis between MIFS and other potential soft-tissue mimics, such as nodular and proliferative fasciitis and inflammatory myofibroblastic tumor. Fine-needle aspiration cytology is a safe and useful tool for the diagnosis of pediatric patients with MIFS and is important for an accurate and precise preoperative workup to optimize subsequent management and treatment.
Related JoVE Video
Role of SDHAF2 and SDHD in von Hippel-Lindau Associated Pheochromocytomas.
World J Surg
Show Abstract
Hide Abstract
Pheochromocytomas (PCCs) develop from the adrenal medulla and are often part of a hereditary syndrome such as von Hippel-Lindau (VHL) syndrome. In VHL, only about 30 % of patients with a VHL missense mutation develop PCCs. Thus, additional genetic events leading to formation of such tumors in patients with VHL syndrome are sought. SDHAF2 (previously termed SDH5) and SDHD are both located on chromosome 11q and are required for the function of mitochondrial complex II. While SDHAF2 has been shown to be mutated in patients with paragangliomas (PGLs), SDHD mutations have been found both in patients with PCCs and in patients with PGLs.
Related JoVE Video

What is Visualize?

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

How does it work?

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

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

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