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Tissue Preparation (tissue + preparation)
Selected AbstractsA Comparison of Four Mohs Tissue Preparation Methods Using Porcine SkinDERMATOLOGIC SURGERY, Issue 9 2010FRCPC, WILLIAM LEAR MD OBJECTIVE Mohs surgery relies on high-quality, rapid tissue preparation and processing. This study evaluated four currently performed tissue preparation and processing methods for speed of processing and depth of cut into the tissue block to achieve a complete high-quality section. METHODS The following four methods were tested: cryoEMBEDDER, float, heat sink, and slide. Standardized specimens of porcine skin were used to ensure uniformity. We measured the time required for a technician to flatten, embed, and cut to the first complete section of each specimen. Additionally, we measured the depth in microns required to cut into an embedded specimen to achieve a complete section. RESULTS There were advantages and disadvantages of each method, and our findings suggest that the heat sink and float methods are more time efficient but that the slide and cryoEMBEDDER methods require less cutting into the specimen to obtain a complete section. The cryoEMBEDDER device used in this study was loaned by cryoEMBEDDER (Salt Lake City, Utah). [source] Noninvasive Imaging, Treatment, and Microscopic Confirmation of Clearance of Basal Cell CarcinomaDERMATOLOGIC SURGERY, Issue 3 2003Mark Goldgeier MD BACKGROUND. The diagnosis of basal cell carcinoma (BCC) is generally established by skin biopsy followed by tissue preparation and microscopic analysis. Treatment of BCC is often accomplished by surgical excision. Objective. To confirm the presence of BCC with a noninvasive imaging technique, to treat the patient with a topical immune response modifier, and to confirm the clearance of BCC noninvasively. METHODS. Confocal microscopy (CM) is a noninvasive technique for real-time imaging of skin in vivo. Imiquimod, an immune response modifier, is applied topically by the patient to the skin lesion. RESULTS. The presence of BCC was confirmed with CM. Posttreatment CM imaging confirmed the clearance of BCC from the entire treatment field. Both the pretreatment and the posttreatment CM findings were confirmed by invasive biopsy. CONCLUSION. The ability to use CM to image in real time without discomfort to the patient makes it a powerful tool to assist in the diagnosis of skin disease. [source] Granular cell tumor of the neurohypophysis: Report of a case with intraoperative cytologic diagnosisDIAGNOSTIC CYTOPATHOLOGY, Issue 1 2008Maria Luisa C. Policarpio-Nicolas M.D. Abstract Cytological techniques including touch and smear preparations are very useful diagnostic modality in the evaluation of central nervous system (CNS) lesions and, in many instances, may be effectively used as the sole modality of tissue preparation for intraoperative consultation. Cytologic preparations offer many advantages over frozen sections for CNS specimens. These include selective examination of multiple areas from small biopsy specimens, superior preservation and details of cellular morphology, fewer artifacts, faster results, and improved cost-effectiveness. We describe the cytologic diagnosis of a granular cell tumor (GCT) of the neurohypophysis in a 33-year-old male who presented with headache and blurred vision. CT scan revealed an enlarged sella with a 2.15 × 2.0 cm pituitary lesion. Transsphenoidal resection of the mass was performed and submitted for intraoperative consultation. Smears and touch preparations were made on a portion of the mass that showed uniform polygonal cells with round to ovoid nuclei and abundant eosinophilic granular cytoplasm. An intraoperative cytological diagnosis of "favor GCT" was rendered. The histologic sections of the remaining material confirmed the diagnosis. Although GCT of the neurohypophysis is very rare, a specific intraoperative cytological diagnosis is possible. We report the clinical, cytological, and pathological findings of a GCT affecting the neurohypophysis. Diagn. Cytopathol. 2008;36:58,63. © 2007 Wiley,Liss, Inc. [source] Endothelin-1 Modulates the Arrhythmogenic Activity of Pulmonary VeinsJOURNAL OF CARDIOVASCULAR ELECTROPHYSIOLOGY, Issue 3 2008AMEYA R. UDYAVAR M.D. Objective: Endothelin-1 has important cardiovascular effects and is activated during atrial fibrillation. Pulmonary veins (PVs) play a critical role in the pathophysiology of atrial fibrillation. The aim of this study was to evaluate whether endothelin-1 affects PV arrhythmogenic activity. Methods: Conventional microelectrodes were used to record the action potentials (APs) and contractility in isolated rabbit PV tissue specimens before and after the administration of endothelin-1 (0.1, 1, 10 nM). The ionic currents of isolated PV cardiomyocytes were investigated before and after the administration of endothelin-1 (10 nM) through whole-cell patch clamps. Results: In the tissue preparation, endothelin-1 (1, 10 nM) concentration dependently shortened the AP duration and decreased the PV firing rates. Endothelin-1 (10 nM) decreased the resting membrane potential. Endothelin-1 (0.1, 1, 10 nM) decreased the contractility and increased the resting diastolic tension. In single PV cardiomyocytes, endothelin-1 (10 nM) decreased the PV firing rates from 2.7 ± 1.0 Hz to 0.8 ± 0.5 Hz (n = 16). BQ-485 (100 ,M, endothelin-1 type A receptor blocker) reversed and prevented the chrono-inhibitory effects of endothelin-1 (10 nM). Endothelin-1 (10 nM) reduced the L-type calcium currents, transient outward currents, delayed rectifier currents, transient inward currents, and sodium,calcium exchanger currents in the PV cardiomyocytes with and without pacemaker activity. Endothelin-1 (10 nM) increased the inward rectifier potassium current, hyperpolarization-induced pacemaker current, and the sustained outward potassium current in PV cardiomyocytes with and without pacemaker activity. Conclusion: Endothelin-1 may have an antiarrhythmic potential through its direct electrophysiological effects on the PV cardiomyocytes and its action on multiple ionic currents. [source] Pharmacological characterization of a novel investigational antimuscarinic drug, fesoterodine, in vitro and in vivoBJU INTERNATIONAL, Issue 8 2008Peter Ney OBJECTIVE To investigate the primary pharmacology of fesoterodine (a novel antimuscarinic drug developed for treating overactive bladder) and SPM 7605 (its active metabolite, considered to be the main pharmacologically active principle of fesoterodine in man) against human muscarinic receptor subtypes, and to investigate in vitro and in vivo functional activity of these agents on the rat bladder compared with existing standard agents. MATERIALS AND METHODS The displacement of radioligand binding by fesoterodine, SPM 7605 and standard agents in membrane preparations of Chinese hamster ovary (CHO) cells expressing the different human muscarinic receptors (M1,M5) was characterized. Agonistic and antagonistic activities were studied using different CHO cell lines stably expressing the human recombinant muscarinic receptor subtypes. The effects of fesoterodine and SPM 7605 on isolated bladder strips contracted by carbachol or electrical field stimulation (EFS) were investigated. In vivo the effects of fesoterodine and SPM 7605 on micturition variables were assessed using continuous cystometry in conscious female Sprague-Dawley rats, and compared to those of oxybutynin and atropine. RESULTS In vitro SPM 7605 potently inhibited radioligand binding at all five human muscarinic receptor subtypes with equal affinity across all five. Fesoterodine had a similar balanced selectivity profile but was less potent than SPM 7605. Both substances were competitive antagonists of cholinergic agonist-stimulated responses in human M1-M5 cell lines and had a similar potency and selectivity profile to the radioligand-binding studies. In rat bladder strips, fesoterodine and SPM 7605 caused a rightward shift of the concentration-response curve for carbachol with no depression of the maximum, and concentration-dependently reduced contractions induced by EFS. The potency of both drugs was similar to that of atropine and oxybutynin. In the presence of the esterase inhibitor neostigmine, the concentration-response curve of fesoterodine was shifted to the right, suggesting that part of the activity was caused by metabolism to SPM 7605 by tissue enzymes. In vivo, low doses (0.01 mg/kg) of fesoterodine and SPM 7605 reduced micturition pressure and increased intercontraction intervals and bladder capacity, but did not affect residual volume. CONCLUSIONS Fesoterodine and its active metabolite, SPM 7605, are nonsubtype selective, competitive antagonists of human muscarinic receptors, but SPM 7605 has greater potency than the parent compound. Pharmacodynamic studies in the rat bladder in vitro confirm the competitive muscarinic antagonist profile of these agents in a native tissue preparation, and in vivo studies in the rat showed effects on bladder function consistent with a muscarinic antagonist profile. [source] Ongoing Nicotinic And Non-Nicotinic Inputs To Inhibitory Neurons In The Mouse ColonCLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 10 2001Andrew K Powell SUMMARY 1. Intracellular microelectrodes were used to record spontaneous and evoked inhibitory junction potentials (IJP) from the circular muscle layer of the mid-distal region of the mouse isolated colon in the presence of nifedipine (1 ,mol/L) and hyoscine (1 ,mol/L). 2. The length of the tissue preparation (> 1 cm) or the presence of the mucosa had no effect on the frequency of spontaneous IJP. 3. Hexamethonium (500 ,mol/L) reduced the frequency of spontaneous IJP to approximately 70% of the control frequency, whereas D -tubocurarine (280 ,mol/L) reduced the frequency to approximately 17% of control. Apamin (250 nmol/L) abolished all spontaneous IJP activity. 4. The greater inhibition of spontaneous IJP in the presence of D -tubocurarine compared with hexamethonium is discussed as a possible ,apamin-like' effect. 5. Although electrically evoked IJP (single pulse at 15 V, 0.6 msec) were not significantly affected by hexamethonium, D -tubocurarine and apamin reduced the amplitude of evoked IJP to approximately 65 and 50% of control, respectively. 6. These results suggest that the properties of spontaneous IJP cannot be inferred by a study of evoked IJP alone. [source] Characterization of the A-type potassium current in murine gastric antrumTHE JOURNAL OF PHYSIOLOGY, Issue 2 2002Gregory C. Amberg A-type currents are rapidly inactivating potassium currents that operate at subthreshold potentials. A-type currents have not been reported to occur in the phasic muscles of the stomach. We used conventional voltage-clamp techniques to identify and characterize A-type currents in myocytes isolated from the murine antrum. A-type currents were robust in these cells, with peak current densities averaging 30 pA pF,1 at 0 mV. These currents underwent rapid inactivation with a time constant of 83 ms at 0 mV. Recovery from inactivation at ,80 mV was rapid, with a time constant of 252 ms. The A-type current was blocked by 4-aminopyridine (4-AP) and was inhibited by flecainide, with an IC50 of 35 ,M. The voltage for half-activation was ,26 mV, while the voltage of half-inactivation was ,65 mV. There was significant activation and incomplete inactivation at potentials positive to ,60 mV, which is suggestive of sustained current availability in this voltage range. Under current-clamp conditions, exposure to 4-AP or flecainide depolarized the membrane potential by 7-10 mV. In intact antral tissue preparations, flecainide depolarized the membrane potential between slow waves by 5 mV; changes in slow waves were not evident. The effect of flecainide was not abolished by inhibiting enteric neurotransmission or by blocking delayed rectifier and ATP-sensitive K+ currents. Transcripts encoding Kv4 channels were detected in isolated antral myocytes by RT-PCR. Immunocytochemistry revealed intense Kv4.2- and Kv4.3-like immunoreactivity in antral myocytes. These data suggest that the A-type current in murine antral smooth muscle cells is likely to be due to Kv4 channels. This current contributes to the maintenance of negative resting membrane potentials. [source] Potential antioxidant activity of celecoxib and amtolmetin guacyl: in vitro studiesAUTONOMIC & AUTACOID PHARMACOLOGY, Issue 1 2007M. Kirkova Summary 1,In vitro studies of the potential antioxidant activity of the selective cyclo-oxygenase-2 inhibitor celecoxib and the non-steroid anti-inflammatory drug amtolmetin guacyl (AMG) were carried out. The study included experiments on the ability of these drugs to affect some indices of the oxidative stress [lipid peroxidation (LP), activity of antioxidant enzymes, glutathione (GSH) level] in rat stomach and colon mucosa and in liver. 2,Celecoxib and AMG did not change the activity of the enzymes GSH-peroxidase, oxidased glutathione (GSSG)-reductase and glucose-6-phosphate-dehydrogenase, as well as the GSH level in all tissue preparations. An increased superoxide dismutase (SOD) activity and a tendency to a decreased Fe/ascorbic acid-induced LP in stomach and colon mucosa were found, but only in the presence of AMG. 3,In the liver, both celecoxib and AMG decreased spontaneous and Fe/ascorbic acid-induced LP. SOD activity was enhanced only in the presence of AMG. 4,Experiments aimed at studying celecoxib and AMG in free oxygen radical-generating systems were also carried out. AMG and tolmetin (the main metabolite of AMG) inhibited OH, -provoked deoxyribose degradation in a Fenton system. Celecoxib had no effect on free radicals when tested in the same system. 5,In conclusion, the results of the present in vitro studies suggest that AMG and celecoxib possess antioxidant and metal-chelating abilities, which might contribute to their beneficial effects. [source] Binding of GTP,[35S] is regulated by GDP and receptor activation.BRITISH JOURNAL OF PHARMACOLOGY, Issue 6 2010Studies with the nociceptin/orphanin FQ receptor Background and purpose:, We have examined the effects of ligand efficacy and receptor density on the binding of guanosine 5,-[,-thio]triphosphate (GTP,S) and GDP to the nociceptin/orphanin FQ (N/OFQ) peptide receptor (NOP)-coupled G-proteins. Experimental approach:, In GTP,[35S] binding experiments, using stable (CHOhNOP) and inducible (CHOINDhNOP) recombinant human and rat NOP we have measured: (i) ligand-specific GDP requirements; (ii) the effects of receptor density on guanine nucleotide affinity/capacity; and (iii) the effect of ligand efficacy on GTP,S association kinetics. Key results:, GTP,S competition curves were shallow and modelled by high- and low-affinity components that were relatively consistent between cell types and tissue preparations. In the presence of 1 µM N/OFQ a high-affinity GDP binding site was also present, but the fraction of total binding was reduced. In an efficacy-dependent manner, the partial agonists [F/G]N/OFQ(1-13)NH2 ([Phe1,(CH2 -NH)Gly2]-nociceptin(1-13)NH2) and naloxone benzoylhydrazone both reduced the fraction of high-affinity sites for GDP (relative to basal). While the pIC50 for high-affinity GDP binding site did not decrease in the presence of 1 µM N/OFQ, N/OFQ produced a significant reduction in pIC50 for the low-affinity site. Agonist-mediated decrease in affinity for GDP binding was efficacy-dependent. GDP displayed three affinities: high, conserved in the presence and absence of ligand; intermediate, present as a low fraction under basal conditions; low (efficacy-dependent), present during receptor activation representing the majority of binding. Conclusions and implications:, The affinity of GTP,[35S] was regulated by GDP and receptor activation caused increased binding of GTP,[35S] through a reduction in GDP affinity. [source] Serum immunoglobulin E levels predict human airway reactivity in vitroCLINICAL & EXPERIMENTAL ALLERGY, Issue 2 2000Schmidt Background Airway hyperresponsiveness to non-specific stimuli is one characteristic feature of airway diseases such as bronchial asthma and chronic bronchitis. Until now, studies aiming to demonstrate a relationship between in vivo conditions associated with airway hyperreactivity and in vitro airway responsiveness have been inconclusive. Objective Since serum immunoglobulin (Ig) E is believed to be one determinant of airway reactivity in vivo, we studied whether in vitro airway reactivity in lung resection material from patients with elevated levels of serum IgE was increased as compared with patients with undetectable IgE. By this approach, we aimed to elucidate the role of circulating IgE for bronchial smooth muscle reactivity in vitro. Methods Bronchial rings from nine patients with total serum IgE levels above 200 U/mL and 10 patients with total serum IgE levels below 10 U/mL were passively sensitized, i.e. incubated overnight with buffer or sensitizing serum containing high levels of total IgE (> 250 U/mL). Afterwards, contractile responses to histamine were assessed in the organ bath. Results Histamine responsiveness was significantly increased in airways obtained from patients with IgE levels above 200 U/mL as compared with airways from patients with IgE levels below 10 U/mL (P < 0.05). Passive sensitization of bronchi from patients with low IgE significantly increased histamine responsiveness, as compared with non-sensitized controls from the same patients (P < 0.05). In contrast, passive sensitization of airways from patients with elevated IgE did not further increase responsiveness. There was no difference in histamine reactivity between non-passively sensitized and passively sensitized tissue preparations from patients with IgE above 200 U/mL and passively sensitized tissues from patients with IgE below 10 U/mL. Conclusion Our findings reveal that elevated levels of serum IgE predict airway hyperresponsiveness to histamine in vitro. At the same time, they indicate that the in vitro model of passive sensitization, in addition to its ability to induce allergen responses, also mimics conditions of non-specific airway hyperreactivity, which are relevant under in vivo conditions. [source] s -CARBOXYMETHYLCYSTEINE INHIBITS CARBACHOL-INDUCED CONSTRICTION OF EPITHELIUM-DENUDED RAT AND HUMAN AIRWAY PREPARATIONSCLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 5-6 2008Dragan Pavlovic SUMMARY 1The effects of s-carboxymethyl-l-cysteine (S-CMC), either administered orally to rats or incubated with tissue preparations from rats and humans, on isometric contractions of tracheal smooth muscle were investigated in the present study using an improved in vitro model of tracheal tube or ring preparations. The involvement of the tracheal epithelium in the observed effects was also investigated. 2The experimental model permitted selective perfusion of the airway tube, luminal-IN or serosal,OUT, and measurement of airway smooth muscle contraction or relaxation in preparations with (+) or without (,) epithelium (Ep), excluding direct effects of airway mucus. 3We found that oral pretreatment of rats with S-CMC (mixed with water; 200 mg/kg per day for 2 weeks), but not short pre-incubation of preparations in vitro (10,3 mol/L S-CMC for 1 h), diminished the sensitivity of ,Ep preparations to carbachol compared with controls (EC50 (,log10 mol/L) values: 5.5 ± 0.1 vs 5.8 ± 0.1, respectively, for IN perfusion (P < 0.005); 5.6 ± 0.1 vs 5.9 ± 0.1, respectively, for OUT perfusion (P < 0.005)), whereas the sensitivity of preparations to aminophylline was not affected. Normal sensitivity to carbachol stimulation was re-established if preparations were pre-incubated with capsaicin. 4It was also found that longer pre-incubation (4 h) of ring-preparations of human bronchus with S-CMC (10,5 mol/L) in vitro resulted in a diminished response to carbachol stimulation. 5In conclusion, S-CMC had small inhibitory effects on the sensitivity of rat and human airway smooth muscle to carbachol, particularly in endothelium-denuded preparations. Whether the epithelium was responding to S-CMC by producing some contracting factor(s) requires further investigation. [source] | ||||||||||