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Whole Tissue (whole + tissue)
Selected AbstractsProtein Kinase C Regulation of Rat Jejunal Transport Systems: Mechanisms Involved in Lactate MovementEXPERIMENTAL PHYSIOLOGY, Issue 6 2002Marisa Tosco We examined whether protein kinase C (PKC) modulates the transport systems involved in lactate movements across the plasma membranes of rat jejunum. In vitro phosphorylated membrane vesicles were used to perform uptake studies, the results of which suggested that PKC activation exerts an inhibitory effect on basolateral H+ -lactate symport, as well as on apical Na+ -glucose cotransport. The specificity of the response to PKC was confirmed by using staurosporine, chelerythrine or 4-,-PMA. Experiments performed using the whole tissue incubated in vitro confirmed the reduction of lactate transport elicited by PKC and gave evidence for an associated inhibition of fluid transport. Na+,K+ -ATPase activity seems to be unaffected by the kinase and inhibited by Ca2+. Taken together, our results suggest that the overall action of PKC results from the simultananeous modulation of multiple pathways, targeted to a reduction of both lactate and bicarbonate transports without altering cell pH homeostasis. [source] MICROWAVE-ASSISTED EXTRACTION OF CAPSAICINOIDS FROM CAPSICUM FRUITJOURNAL OF FOOD BIOCHEMISTRY, Issue 2 2004OPAL J. WILLIAMS The applicability of microwave irradiation to assist the extraction of capsaicinoids from capsicum fruit was investigated. The procedure involved irradiation of 2 g samples in a closed-vessel followed by gas chromatography of capsaicinoid derivatives. The optimum conditions for extraction were determined to be acetone at 30% power for 7 min irrespective of ground or whole tissue. The yield of the compounds extracted was significantly greater (P < 0.05) using microwave-assisted extraction (MAE) compared to traditional reflux and shaken flask methods. A single extraction step was efficient in recovering approximately 95% of the total capsaicinoid fraction in 15 min compared with 2 h for the reflux and 24 h for the shaken flask methods. Due to the considerable savings in time and energy as well as reliability, this technique is suitable for fast extraction of capsaicinoids from large samples. [source] Quantification of the graphical details of collagen fibrils in transmission electron micrographsJOURNAL OF MICROSCOPY, Issue 1 2001Y. Xia A novel 2D image analysis technique is demonstrated. Using the digitized images of articular cartilage from transmission electron microscopy (TEM), this technique performs a localized ,vector' analysis at each region that is large enough to include several or tens of collagen fibrils but small enough to provide a fine resolution for the whole tissue. For each small and localized region, the morphology of the collagen fibrils can be characterized by three quantities essential to the nature of the tissue: the concentration of the fibrils, the overall orientation of the fibrils, and the anisotropy of the fibrils. This technique is capable of providing new insight to the existing technology by assigning quantitative attributes to the qualitative graphics. The assigned quantities are sensitive to the fine structure of the collagen matrix and meaningful in the architectural nature of the collagen matrix. These quantities could provide a critical linkage between the ultrastructure of the tissue and the macroscopic behaviours of the material. In addition, coarse-graining the microscopic resolution of EM without compromising the essential features of the tissue's structure provides a direct view of the tissue's morphology and permits direct correlations and comparisons among interdisciplinary techniques. [source] Molecular Classification of Thyroid Nodules by Cytology,,§THE LARYNGOSCOPE, Issue 4 2008Nitin A. Pagedar MD Abstract Objectives: Fine needle aspiration (FNA) biopsy of thyroid nodules provides cytologic specimens whose interpretation can direct patients toward either thyroidectomy or observation. Approximately 20% of FNA specimens yield an indeterminate result. Recent studies have characterized differences in gene expression between benign and malignant conditions, most often using whole tissue. Our goal was to determine the feasibility of quantitative polymerase chain reaction (qPCR)-based gene expression analysis in cytologic samples. For five genes shown to be over-expressed in thyroid carcinomas (fibronectin, galectin-3, Met/HGFR, MUC1, and GA733-precursor), we compared expression among pathologic states. Study Design: Prospective laboratory analysis of 20 thyroidectomy specimens. Methods: Routine microscopy was performed. Cytologic samples were obtained from the dominant nodules, and RNA was extracted. Preliminary analysis using fluorometry and reverse-transcriptase (RT)-PCR was performed. Expression levels of the test genes in nodules and from control samples were measured by real-time qPCR. Fold changes in gene expression were compared. Results: Only one specimen did not yield sufficient intact RNA for gene expression analysis. RT-PCR revealed satisfactory RNA recovery in all other specimens. qPCR showed significant over-expression of fibronectin in the papillary carcinomas compared with the goiters (P = .0013), follicular adenomas (P = .0014), and follicular carcinomas (P = .0001). Differences in both fibronectin and MUC1 expression between the follicular carcinomas and the follicular adenomas were also significant (P = .025 and .045, respectively). Conclusions: Cytologic specimens were a satisfactory source of tissue for qPCR-based gene expression analysis. Both fibronectin and MUC1 were differentially expressed in follicular adenomas and follicular carcinomas, and fibronectin expression differed in papillary carcinomas compared with the other lesions. These results may form the basis of a clinical predictor for lesions with indeterminate or suspicious cytology. [source] Phospholipid profile of rat testis, its unique high level of monolysocardiolipin and its lipolytic capabilities in vitro.CELL BIOCHEMISTRY AND FUNCTION, Issue 4 2008A chromatographic analysis Abstract The phospholipid profiles of testes and heart of 1-, 3-, and 6-month-old rats and their in vitro response to the endogenous phospholipases at pH 7.4 and 38°C for 60,min were analyzed by TLC technology and densitometry. A noticeable high level of monolysocardiolipin (MLCL) was shown in rat testes of all samples analyzed (1-, 3-, and 6-month-old), both control and incubated. In contrast, rat heart control samples revealed a high level of CL and no MLCL was detected. MLCL was only produced subsequent to in vitro incubation of whole tissue homogenate at pH 7.4 and 38°C for 60,min, with concurrent reduction of CL. Alkenyl phosphatidyl ethanolamine (PE) was the major plasmalogen of rat testes. Following in vitro incubation, (a) a very low level of lyso PE plasmalogen was produced only in 3- and 6-month-old rat testes, (b) ceramide was also produced in all testes analyzed with concurrent reduction of sphingomyelin indicating the action of sphingomyelinase. These data clearly illustrate, for the first time, the presence of high levels of MLCL in all rat testes studied which probably is related to the physiological activity in vivo and requires further investigation. Copyright © 2008 John Wiley & Sons, Ltd. [source] Studies on the endogenous phospholipids of chick embryo myocardium and their in vitro hydrolysis by endogenous phospholipases during embryogenesisCELL BIOCHEMISTRY AND FUNCTION, Issue 5 2007Fatma M. Helmy Abstract The phospholipid profiles of the myocardium (from 10- and 18-day old chick embryos and 13-day old chick) and their in vitro response to the endogenous lipolytic enzymes (mainly of the phospholipase group) at pH 7.4 and 38°C for 60,min were analyzed by TLC technology and densitometry. Cardiolipin (CL) was shown to be one of the major phospholipids of the chick embryo myocardium and its concentration increased as the chick embryo advanced in development. Monolysocardiolipin (MLCL) was produced subsequent to in vitro incubation of whole tissue homogenates in all myocardia studied as well as a concurrent reduction in CL. This deacylation of CL increased in magnitude as the chick embryo advanced in development indicating its age relatedness. The level of phosphatidyl ethanolamine (PE) plasmalogen was also high in all myocardia studied. Lyso alkenyl PE (LPE) was produced subsequent to in vitro incubation and its level increased as the chick embryo advanced in development, indicating PLA2 action on the sn-2 fatty acid of PE. Phosphatidyl choline (PC) plasmalogen was also present in the chick embryo myocardium and its level increased gradually as the chick embryo advanced in development. In contrast, yolk-sac membrane contains very minute amounts of CL and PE. No PC was detected and no LPE was formed following in vitro incubation. The yolk of the unfertilized chicken egg has no CL and has very minute amounts of PE, no PC and no lysophospholipids were detected following in vitro incubation in all samples analyzed. Copyright © 2007 John Wiley & Sons, Ltd. [source] 2-Amino-1-methyl-6-phenylimidazo[4,5- b]pyridine (PhIP),induced mutagenesis in cultured Big BlueÔ rat mammary epithelial and fibroblast cellsENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 2-3 2002Heather M. McDiarmid Abstract Epithelial cells are the primary site of carcinogenesis in most tissues, including the mammary gland. As an alternative to the study of mutation induction in whole tissues in vivo, we have established Big BlueÔ transgenic rat cell lines from the mammary epithelium (BBR/ME) and the mammary stroma (BBR/MFib), to permit a comparison of their mutagenic responses to carcinogens. We previously demonstrated their responsiveness to the alkylating agent N -ethyl- N -nitrosourea (ENU) (McDiarmid H et al. [2001]: Mutat Res 497:39,47). Here, we examined the responses of cultured epithelial and stromal cells to the protein pyrolysis product and mammary carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5- b]pyridine (PhIP). Rat hepatic S9 was used as a source of bioactivation enzymes. Mutant induction (cII locus) and clonogenic survival were measured as a function of PhIP concentration. PhIP mutagenicity was observed in the fibroblast cells, but the greater toxicity of PhIP to the epithelial cells prevented a definitive evaluation of mutagenicity. Since PhIP may be detoxified by conjugation with glutathione, we measured glutathione levels and glutathione- S -transferase expression and activities in both cell lines. The epithelial cells had higher glutathione- S -transferase enzyme activity and protein expression than did the fibroblast cell line. Because the epithelial cells were more sensitive to toxicity, glutathione conjugation evidently plays only a minor role in PhIP toxicity and mutagenicity in our cell lines. Environ. Mol. Mutagen. 39:245,253, 2002. © 2002 Wiley-Liss, Inc. [source] 4411: Immunohistochemical methods to evaluate vitreoretinal scaringACTA OPHTHALMOLOGICA, Issue 2010ML BOCHATON-PIALLAT Purpose Formation of scarlike epiretinal membranes (ERMs) constitutes potentially the end stage of evolution of proliferative vitreoretinopathy (PVR), proliferative diabetic retinopathy (PDR) and idiopathic vitreoretinopathy. Among various cellular populations, ERMs contain cells with contractile features typical of myofibroblasts. Myofibroblasts have been described in granulation tissue during wound healing and in practically all fibrocontractive diseases, in which they participate in the generation of isometric tension and in the synthesis of extracellular matrix components; these phenomena are in turn responsible for granulation tissue remodeling and retraction. The main marker of the myofibroblastic phenotype is the expression of alpha-SMA. The transforming growth factor-beta1 and the ED-A splice variant of cellular fibronectin, an extracellular matrix component, are key players of the complex process of myofibroblast differentiation. Methods Proteins were detected by means of immunohistochemical staining on paraffin sections from formol fixed tissues and double immunofluorescence staining on whole tissues. Samples were observed by using classical light and confocal microscopes. Results The presence of alpha-SM actin-positive myofibroblasts was associated with the expression of TGF-beta1, TGF-beta receptor II, and ED-A FN in all types of ERMs studied. Conclusion The results furnish new data on the mechanism of alpha-SM actin stimulation in fibroblasts in a human pathologic setting. [source] | |||||||||||||