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At least 253 records · Page 14Linked to original sources
PubMed2005

Gene expression analysis in sections and tissue microarrays of archival tissues by mRNA in situ hybridization.

Altered expression of genes in diseased tissues can prognosticate a distinct natural progression of the disease as well as predict sensitivity or resistance to particular therapies. Archival tissues from patients with a known medical history and treatments are an invaluable resource to validate the utility of candidate genes for prognosis and prediction of therapy outcomes. However, stored tissues with associated long-term follow-up information typically are formalin-fixed, paraffin-embedded specimen and this can severely restrict the methods applicable for gene expression analysis. We report here on the utility of tissue microarrays (TMAs) that use valuable tissues sparingly and provide a platform for simultaneous analysis of gene expression in several hundred samples. In particular, we describe a stable method applicable to mRNA expression screening in such archival tissues. TMAs are constructed from sections of small drill cores, taken from tissue blocks of archival tissues and multiple samples can thus be arranged on a single microscope slide. We used mRNA in situ hybridization (ISH) on >500 full sections and >100 TMAs for >10 different cDNAs that yielded >10,000 data points. We provide detailed experimental protocols that can be implemented without major hurdles in a molecular pathology laboratory and discuss quantitative analysis and the advantages and limitations of ISH. We conclude that gene expression analysis in archival tissues by ISH is reliable and particularly useful when no protein detection methods are available for a candidate gene.

Animals↗
PubMed1991

17 Beta-hydroxysteroid dehydrogenases in human breast tissues: purification and characterization of soluble enzymes and the distribution of particulate and soluble forms in adipose, non-adipose and tumour tissues.

17 Beta-Hydroxysteroid dehydrogenase (17 beta-HSD) is present in multiple forms in human breast tissue. One soluble form, with a molecular weight of approximately 35 kDa, was purified to near homogeneity from whole normal breast tissue. This form catalysed the oxidation of oestradiol and the reduction of oestrone, with NADP+ and NADPH as the preferred coenzymes. Three other soluble forms with higher molecular weights (in the range 50-80 kDa) were isolated. They catalysed the oxidation of oestradiol but not the reduction of oestrone, and all of them had properties very different from those of the low molecular weight enzyme. Activities of 17 beta-HSD were measured in particulate and soluble fractions from normal breast adipose and non-adipose tissues, and from breast tumours obtained from post-menopausal women, in the oxidative direction with NAD+ and NADP+ as coenzymes and in the reductive direction with NADH and NADPH as coenzymes. Particulate fractions from tumours had much higher oxidative and reductive activities than those from normal tissues. Soluble fractions from tumours had higher oxidative activities than those from the normal tissues but similar reductive activities. The major soluble form of 17 beta-HSD in adipose tissue was the 35 kDa enzyme which had both oxidative and reductive activities. In contrast, the majority of the soluble activity in non-adipose tissue was due to enzymes, with molecular weights in the range 50-80 kDa, which had oxidative activity only. The soluble fractions of tumours, like those of non-adipose tissue, contained enzymes with molecular weights in the range 50-80 kDa. In addition, they contained a 35 kDa enzyme with properties different from those of the enzyme with the same molecular weight present in adipose tissue.

17-Hydroxysteroid Dehydrogenases↗
PubMed2004

Detection of soft tissue foreign bodies in the presence of soft tissue gas.

OBJECTIVE: To determine the effect of soft tissue gas on the accuracy of foreign body detection by realtime sonography. METHODS: This was a prospective randomized study using glass, metal, and bone inserted into turkey breasts to simulate human soft tissue foreign bodies. Air was subsequently injected around a random selection of the foreign bodies to simulate soft tissue gas that can accompany a blast or high-force injury. Using a linear transducer, physicians credentialed in the use of sonography were each asked to scan the breasts, identify the location of any foreign body, and describe whether the object located was bone, metal, or glass. They were also asked to describe the characteristics of the foreign body, including surface echogenicity, visibility, and artifacts, if any. RESULTS: The sensitivity for localization of each foreign body by each sonographer was 100% (48 of 48) and was unaffected by the presence of soft tissue gas. The accuracy of classifying the foreign body was poor except with bone. Glass and metal were often confused with each other. With the addition of soft tissue gas over the foreign bodies, the sensitivity of classifying the foreign body was decreased further from a combined 58% to 28%. The presence of soft tissue gas decreased the amount of reflection of the foreign body and obscured the subtle differences in the brightness of each foreign body, leading to a decrease in the accuracy of identification but not localization of the foreign body. CONCLUSIONS: In an experimental model, soft tissue gas does not affect the localization of soft tissue foreign bodies. However, correct identification of the type of foreign body is limited by soft tissue gas because of loss of the typical sonographic characteristics.

Animals↗
PubMed2006

Effect of controlled volumetric tissue heating with radiofrequency on cellulite and the subcutaneous tissue of the buttocks and thighs.

BACKGROUND: Regardless of diet and exercise, genetics plays an important part in creating puckering skin or dimples, which are difficult to hide at any age. The demand for a nonsurgical, noninvasive treatment of cellulite has inspired some manufacturers to invest in a new age of sophisticated devices and treatment therapies to repair the skin and improve contours. Although many of these new choices have demonstrated a smoothing effect (following a multitude of treatments), the objective documentation has in most cases been limited to biopsies, circumference measurements, and photographic evidence. HYPOTHESIS: We believe that the application of noninvasive high-energy radiofrequency (RF) to the skin of the thigh and buttocks heats the subcutaneous adipose tissue, causing collagen fibers to contract. The resulting impact to the subcutaneous tissue and collagen is expected to improve the skin's external architecture. Given that the subcutaneous tissue and adipose tissue are difficult to evaluate through histological methods, this investigation seeks to demonstrate the changes that occur when applying 2 treatments of high-energy RF on the subcutaneous tissue of thighs and buttocks utilizing real-time ultrasound image scanning. MATERIALS AND METHODS: Twenty-six healthy female patients (ages 18 to 50) with visible bilateral cellulite (grade 1 to 3) on either the buttocks and/or thighs received 2 treatment sessions (15 days apart) of unipolar RF using the Accent RF System (Alma Lasers Inc). The system utilizes a unipolar RF applicator that is electrically cooled to aid in patient comfort during the treatment. Appropriate energy was set and the treatment was delivered in 3 passes of 30 seconds each. Evaluation of the thickness of the subcutaneous tissue on buttocks and thighs took place before the first treatment, second treatment, and 15 days following the second treatment with a with real-time scanning image ultrasound (Philips Medical Systems). Clinical improvement was objectively evaluated through comparative pre- and post-treatment measurements of the distance between the stratum corneum to the Camper's fascia and from the stratum corneum to the muscle. The study also evaluated the structure and changes of the collagen (thickening and realignment of septae) resulting from 2 treatments of RE Photography was used to document contour and superficial changes. RESULTS: From the measurements of the distance between the stratum corneum to the Camper's fascia and from the stratum corneum to the muscle we were able to demonstrate that 68% of the patients presented a contraction of the volume of approximately 20%. CONCLUSIONS: Based on the demonstrated results with real-time ultrasound scanning, we have observed that 2 RF treatments on the subcutaneous tissue of the buttocks and thighs provide a volumetric contraction effect in the majority of patients. This validates the primary hypothesis of our protocol and establishes that the RF energy works on the connective tissue of the subcutaneous adipose tissue. This effect should be the same on any other body part.

Adolescent↗
PubMed1990

In situ distribution of the beta-subunit of platelet-derived growth factor receptor in nonneoplastic tissue and in soft tissue tumors.

The distribution of the beta-subunit of platelet-derived growth factor receptor (PDGFR-beta) was assessed by a sensitive immunoalkaline phosphatase technique using the monoclonal antibody PR7212. Frozen tissue sections of several nonneoplastic human tissues were stained along with 42 soft tissue sarcomas, 16 benign soft tissue proliferations, and 7 epithelial tumors. In all nonneoplastic tissue, there was intense labeling of cell processes of perivascular fibroblasts or pericytes in and about the walls of muscular blood vessels and of fibroblast cell processes around some glandular and ductal epithelia. No PDGFR-beta was found in the endothelial cells of muscular arteries and veins, but cells of uncertain identity within some capillaries were immunoreactive and the possibility that endothelial cells of some small capillaries express PDGFR-beta could not be excluded. In kidney there was strong labeling of glomerular mesangial cells and interstitial fibroblasts. Some histological types of soft tissue sarcomas were uniformly and strongly labeled with anti-PDGFR-beta, but other types were infrequently labeled or unreactive. The order of decreasing frequency and strength of labeling of the various types of benign and malignant soft tissue proliferations was as follows: benign fibromatosis and neurofibroma greater than malignant fibrous histiocytoma greater than liposarcoma greater than leiomyosarcoma greater than rhabdomyosarcoma. No tumor cell labeling was detected in epithelioid, synovial or clear cell sarcomas, leiomyomas, or carcinomas, but there was usually strong labeling of fibroblast and/or pericyte cell processes within tumor, especially around blood vessels. We conclude that PDGFR-beta is strongly expressed by vascular and stromal tissues of most tumors and normal organs and by tumor cells of several types of soft tissue tumors and proliferations, most notably those of fibroblastic origin.

Blood Vessels↗
PubMed1981

Antibiotic concentrations in blood and tissue. Intraoperative ischemia as a model for the determination of tissue concentrations using mezlocillin and oxacillin as examples.

Blood present in the tissue distorts the results obtained during measurement of antibiotic tissue levels. The greater the blood supply to the tissue, the bigger the error when tissue levels are measured. The examination of tissue removed during temporary intraoperative ischemia is an alternative to the indirect mathematical method of correcting errors of blood level measurement. This technique was tested by determining tissue concentrations after i.v. injection of 4 g mezlocillin and 2 g oxacillin. The tissue was removed during operations on limbs after the application of an Esmarch's bandage. Determination of hemoglobin in ischemic tissue confirmed exsanguination so that the 6-((R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenylacetamido )-penicillanic acid-Na (mezlocillin) and 5-methyl-3-phenyl-4-isoxazolylpenicillin (oxacillin) concentrations determined corresponded to the actual tissue levels. The concentrations of mezlocillin and oxacillin in fat, skin, tendon, muscle, ganglion, aponeurosis, tumour, ligament, subcutis, synovial fluid, epineurium and serum are given in the text.

Adult↗
PubMed2006

Langerhans cells express matrix metalloproteinases 9 and 2 and tissue inhibitors of metalloproteinases 1 and 2 in healthy human gingival tissue and in periodontitis.

BACKGROUND: As antigen-presenting cells, Langerhans cells may play an important role in the initiation and maintenance of periodontal disease. This study is the first report that extends our knowledge of the expression of matrix metalloproteinases and their endogenous tissue inhibitors by Langerhans cells in healthy and diseased gingival tissues. METHODS: Single and double immunolabeling procedures were carried out using monoclonal antibodies against CD1a, matrix metalloproteinases 2 and 9, and tissue inhibitors of matrix metalloproteinases 1 and 2, and analyzed by conventional and confocal microscopes. RESULTS: Langerhans cells expressed matrix metalloproteinases 2 and 9, and tissue inhibitors of matrix metalloproteinases 1 and 2 in healthy and diseased gingival tissues. The tissue inhibitors of matrix metalloproteinase-positive Langerhans cells were mainly observed in the upper epithelial layers. Matrix metalloproteinase 9-positive Langerhans cells were observed especially during periodontitis and in the basal epithelial layer or crossing the basement membrane. CONCLUSION: During periodontal disease, changes in the expression of matrix metalloproteinases and their tissue inhibitors by gingival Langerhans cells could be implicated in the migration of the cells towards the connective tissue.

Cell Movement↗