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Changes in the lipogenic response to feeding of liver, white adipose tissue and brown adipose tissue during the development of obesity in the gold-thioglucose-injected mouse.

Lipogenic response to feeding was measured in vivo in liver, epididymal white adipose tissue (WAT) and interscapular brown adipose tissue (BAT), during the development of obesity in gold-thioglucose (GTG)-injected mice. The fatty acid synthesis after a meal was higher in all tissues of GTG-treated mice on a total-tissue basis, but the magnitude of this increase varied, depending on the tissue and the time after the initiation of obesity. Lipogenesis in BAT from GTG mice was double that of control mice for the first 2 weeks, but subsequently decreased to near control values. In WAT, lipogenesis after feeding was highest 2-4 weeks after GTG injection, and in liver, lipid synthesis in fed obese mice was greatest at 7-12 weeks after the induction of obesity. The post-prandial insulin concentration was increased after 2 weeks of obesity, and serum glucose concentration was higher in fed obese mice after 4 weeks. These results indicate that increased lipogenesis in GTG-injected mice may be due to an increase in insulin concentration after feeding and that insulin resistance (assessed by lipogenic response to insulin release) is apparent in BAT before WAT and liver.

Adipose Tissue↗

Application of tissue microarray: evaluation of the expression of S-100-positive dendritic cells, tumor suppressor gene p63 and tissue inhibitor of metalloproteinase-1 in laryngeal carcinoma.

OBJECTIVE: To evaluate the predictive role of S100-positive dendritic cells, tissue inhibitor of metalloproteinase-1 (TIMP1) and p63 gene in laryngeal carcinoma using a tissue chip. MATERIAL AND METHODS: The expression of dendritic cells, TIMP1 and p63 gene in a series of 85 primary laryngeal carcinoma patients who had undergone operations in our hospital between 1992 and 2000 was studied immunohistochemically using the streptavidin-biotin peroxidase-conjugated method. RESULTS: Some cases were lost or showed no tumor tissue in the tissue chip. The rate of expression of dendritic cells was 59.5% and there were significant differences in expression between the differential degrees of laryngeal squamous carcinoma and the different clinical stages (p <0.05). The basal membrane and extracellular matrix of the specimens were strongly stained by TIMP1 in 55.7% of cases. There were statistically significant correlations between TIMP1 protein expression and early- and advanced-stage tumors, lymph node metastasis and 3-year survival rate (p <0.05). No statistically significant correlations were found between p63 protein expression and any of the clinical parameters. CONCLUSIONS: The tissue microarray technique used in this study is quick, inexpensive and very consistent. It is suggested that TIMP1 and S100 should be used as clinical discriminators of laryngeal carcinoma.

Adult↗

Distribution of a major connective tissue protein, fibronectin, in normal human tissues.

Fibronectin is a major surface-associated glycoprotein of cultured fibroblasts and it is also present in human plasma. Antiserum specific for human fibronectin was used to study the distribution of fibronectin in normal adult human tissues. The protein was detected (a) characteristically in various basement membranes including capillary walls: (b) around individual smooth muscle cells and in the sarcolemma of striated muscle fibers; and (c) in the stroma of lymphatic tissue and as thin fibers in loose connective tissue. The distribution of fibronectin was distinct from that of collagen and elastic fibers, but was very similar to reticulin, as demonstrated by conventional histological staining. The results indicate that fibronectin is a major component of connective tissue matrix. The distribution also indicates that most types of adherent cells abut fibronectin-containing structures. This supports the possible role of fibronectin in cell-cell and cell-matrix interactions in tissues.

Basement Membrane↗

Novel cell-scaffold interactions encountered in tissue engineering: contractile behavior of musculoskeletal connective tissue cells.

Methods employed in the course of tissue engineering often offer unique opportunities to observe cell-matrix interactions that cannot otherwise be viewed. These observations may provide insights into cell behavior than can contribute important new knowledge about cell biology. One such set of observations led to the discoveries that musculoskeletal connective tissue cells express a contractile muscle actin isoform, alpha-smooth muscle actin, and can contract. This knowledge may help to explain how these cells generate forces required for certain physiological and pathological functions, and this information may inform future approaches to regulate this function to advance tissue engineering. Tissue engineering science is thus emerging as an importance force that can both contribute to cell and molecular biology and add to the fund of knowledge supporting the production of tissue in vitro or in vivo to improve the management of a wide variety of disorders.

Actins↗

Immunolocalization of tissue inhibitor of metalloproteinases-1 within ovine periovulatory follicular and luteal tissues.

In previous studies, tissue inhibitor of metalloproteinases (TIMP)-1 mRNA increased in follicular tissue after the preovulatory gonadotropin surge and was expressed in luteal tissue. However, the localization of TIMP-1 protein within ovine periovulatory follicular and luteal tissues is unknown. The objectives of the present study were to 1) localize TIMP-1 within follicles collected before and after a preovulatory gonadotropin surge and within Day 3 and Day 10 corpora lutea (CL), 2) determine whether TIMP-1 was colocalized to Day 10 luteal cells with oxytocin or TIMP-2, and 3) determine whether TIMP-1 was present within secretory granules of large luteal cells. Ovaries were removed from ewes before (presurge; n = 4) or 12-14 h after (postsurge; n = 5) an LHRH-induced gonadotropin surge (36 h following PGF2 alpha-induced luteolysis; objective 1). Additionally, ovaries containing CL were collected on Days 3 (n = 5; objective 1) and 10 (n = 4, 3, and 2 for objectives 1, 2, and 3, respectively). TIMP-1 immunoreactivity was observed within the granulosa cells of postsurge but not presurge follicles. On Days 3 and 10, TIMP-1 was localized within luteal tissue in a cell-specific manner. On Day 10, many of the cells that were immunopositive for TIMP-1 were judged to be large luteal cells on the basis of morphology (diameter > 22 microns; round nucleus) and colocalization with oxytocin and TIMP-2. Electron microscopy demonstrated that TIMP-1 was localized to secretory granules undergoing exocytosis from Day 10 large luteal cells. These data indicate that TIMP-1 is produced by granulosa cells following a gonadotropin surge and is packaged in secretory granules by large steroidogenic cells of the ovine CL.

Animals↗

The beginning of a new era in tissue expansion: self-filling osmotic tissue expander--four-year clinical experience.

The osmotic tissue expander is a new device made of a hydrogel expanding skin that does not require external fillings. Once implanted, it absorbs body fluids, which leads to a gradual swelling of the device. The swelling phase is completed in 6 to 8 weeks and results in skin gain. Different shapes and sizes are available, and the devices can be used in almost every area of the body. Over a 4-year period, the osmotic tissue expander was used in 58 patients in different areas of the body. A round osmotic tissue expander was mainly used in breast reconstruction, and a rectangular expander was used for defect coverage after excision (i.e., of scars and tumors). The mean age of the patients was 49.34 years (range, 4 to 76 years). During the expansion phase, the patients noted only a little discomfort and pain for the first few days. Without a silicone membrane in the first-generation expander, the rate of successful explantation and good final result was 81.5 percent. In a few cases, rapid swelling of the device led to the introduction of a silicone membrane that encloses the expander and leads to a slower, more gradual, and consistent swelling. After introduction of the silicone envelope, the success rate improved to 91 percent. The expander is now used with a silicone membrane in every case. The osmotic tissue expander has many advantages compared with the conventional expander: there is no need for painful external fillings and the risk of external infections is avoided. The expander is 10 percent of its final volume and only requires a short incision and a small pocket. An operation can easily be performed under local anesthesia, with minimal tissue mobilization in older children and compliant patients.

Adolescent↗

Neural tissue xenotransplantation: what is needed prior to clinical trials in Parkinson's disease? Neural Tissue Xenografting Project.

Embryonic allografted human tissue in patients with Parkinson's disease has been shown to survive and ameliorate many of the symptoms of this disease. Despite this success, the practical problems of using this tissue coupled to the ethical restrictions of using aborted human fetal tissue have lead to an exploration for alternative sources of suitable material for grafting, including xenogeneic embryonic dopaminergic-rich neural tissue. Nevertheless, xenografted neural tissue itself generates a number of practical, ethical, safety, and immunological issues that have to be addressed prior to any clinical xenotransplant program. In this article we review these critical issues and set out the criteria that we consider need to be met in the development of our clinical xenotransplantation research programs. We advocate that these, or similar, criteria should be adopted and made explicit by other centers contemplating similar clinical trials.

Animals↗

Myxoid tissue: its morphology, histochemistry, and relationship with other supporting tissues.

Myxoid tissue was studied in the supporting organ of the cat epiglottis ("epiglottic cartilage"). Under the light microscope, myxoid tissue was characterized by stellate cells placed into an avascular acidic extracellular matrix. This extracellular matrix was alcianophilic at pH = 2.5, reacting with the colloidal iron stain, and staining metachromatically with toluidine blue O at pH = 5.0. Treatment of sections with testicular hyaluronidase abolished these reactions. In addition, staining persisted after methylation/saponification pretreatment, indicating hyaluronic acid as the main acidic component of myxoid extracellular matrix. Under the electron microscope, myxoid extracellular matrix formed flocculent electron dense precipitates. Stellate myxoid cells were characterized by bundles of intermediate (8 nm) cytoplasmic filaments. Myxoid cells were devoid of a basal lamina, contained a few small lipid droplets, and stored some glycogen. Bundles of collagen fibrils, 80-120 nm in diameter, were seen in myxoid areas. Myxoid cells reacted to S-100 protein, glial fibrillary acidic protein, and neuron specific enolase. Moreover, in adult animals, myxoid cells stained for neurofilament protein 200. All these markers were also present in chondrocytes of elastic and fibrous cartilage, indicating a close relationship between myxoid cells and chondrocytes. This was supported by the observation of continuous transitional forms of myxoid tissue into elastic or fibrous cartilage. In 8-week-old kittens, the supporting organ of the epiglottis was found mainly to consist of myxoid tissue with only a few interspersed islets of chondrocytes. It is therefore concluded that myxoid tissue can serve as a precursor of cartilage.

Adipose Tissue↗

Subpedicle connective tissue graft versus guided tissue regeneration with bioabsorbable membrane in the treatment of human gingival recession defects.

The purpose of the present clinical study was to evaluate the effect of guided tissue regeneration (GTR) in comparison to subpedicle connective tissue graft (SCTG) in the treatment of gingival recession defects. A total of 12 patients, each contributing a pair of Miller's Class I or II buccal gingival recessions, was treated. According to a randomization list, one defect in each patient received a polyglycolide/lactide bioabsorbable membrane, while the paired defect received a SCTG. Treatment effect was evaluated 6 months postsurgery. Clinical recordings included full-mouth and defect-specific oral hygiene standards and gingival health, recession depth (RD), recession width (RW), probing depth (PD), clinical attachment level (CAL), and keratinized tissue width (KT). Mean RD significantly decreased from 3.1 mm presurgery to 1.5 mm at 6 months postsurgery for the GTR group (48% root coverage), and from 3.0 mm to 0.5 mm for the SCTG group (81% root coverage). RD reduction and root coverage were significantly greater in SCTG group compared to GTR group. Mean CAL gain amounted to 1.7 mm for the GTR group, and 2.3 mm in the SCTG group. No significant differences in PD changes were observed within and between groups. KT increased significantly from presurgery for both treatment groups, however gingival augmentation was significantly greater in the SCTG group compared to GTR group. Results indicate that: 1) treatment of human gingival recession defects by means of both GTR and SCTG procedures results in clinically and statistically significant improvement of the soft tissue conditions of the defect; and 2) treatment outcome was significantly better following SCTG compared to GTR in terms of recession depth reduction, root coverage, and keratinized tissue increase.

Absorbable Implants↗

Comparative clinical study of guided tissue regeneration with a bioabsorbable bilayer collagen membrane and subepithelial connective tissue graft.

BACKGROUND: During the last decade, there have been great strides in the treatment of gingival recession defects, especially with subepithelial connective tissue graft and guided tissue regeneration (GTR) procedures. Gingival recession represents a significant concern for patients. It is necessary to choose the most appropriate procedure in order to obtain more root coverage while avoiding clinical disadvantages. The purpose of this randomized clinical trial was to evaluate the use of a bioabsorbable bilayer collagen membrane with GTR compared to a connective tissue graft in the treatment of gingival recession defects. METHODS: Twenty patients each contributing a pair of Miller Class I or II buccal gingival recessions were treated. In each pair, one recession was randomly assigned for treatment with GTR using a bioabsorbable bilayer collagen membrane and the other treated with subepithelial connective tissue graft (CTG). Clinical measurements taken at baseline (D0) and 3 and 6 months post-treatment included recession depth (RD), recession width (RW), probing depth (PD), and clinical attachment level (CAL). RESULTS: Data were analyzed using the non-parametric Wilcoxon matched pair test. All results were statistically significant. Both treatments resulted in a significant gain of root coverage (P<0.0001), amounting to an average of 2.80 mm at 3 months in the GTR group and 3.34 mm in the CTG group. At 6 months, the decrease of the mean RD remained statistically significant: 2.70 mm (74.59% root coverage) in the GTR group and 3.19 mm (84.84% root coverage) in the CTG group. The mean RW also decreased from 4.48 mm at D0 to 2.42 mm at 6 months in the GTR group, and from 4.38 mm at D0 to 1.35 mm at 6 months in the CTG group, representing a percentage of coverage of 45.98% and 69.18%, respectively. Mean CAL gain obtained between D0 and 6 months with the GTR procedure and CTG was 3.31 mm and 3.09 mm, respectively, and was significant within groups. At 3 and 6 months, the differences in the results for RD, CAL, and RW were not statistically significant between the 2 groups. However, the difference was significant for PD at 3 and 6 months. CONCLUSIONS: The results suggest that a bioabsorbable bilayer collagen membrane can be used in the GTR treatment of human buccal recession defects, with no statistically significant differences between this procedure and connective tissue grafts.

Absorbable Implants↗

[Expression and significance of tissue inhibitors of metallproteinases-1 and -2 in serum and liver tissue of patients with liver cirrhosis].

OBJECTIVE: To investigate the expression and location of tissue inhibitors of metallproteinases-1 and -2 (TIMP-1 and TIMP-2) in liver tissue of patients with cirrhosis and to investigate the correlation between TIMPs in liver and those in serum. METHODS: TIMP-1 and TIMP-2 in the serum of 106 patients with liver cirrhosis were detected by solid-phase absorption to sensitized erythrocytes (SPASE). The expression of TIMP-1 and TIMP-2 related antigens in liver biopsy specimens from 30 patients with liver cirrrhosis was detected by immunohistochemical method with specific monoclonal antibody (McAb), and the results thus obtained were compared with the results of the seral TIMPs from the same patients. 10 normal livers were used as controls. RESULTS: The positive rates of TIMP-1 and TIMP-2 in the 106 serum samples of patients with cirrhosis were 73.58% and 61.32% respectively. TIMP-1 and TIMP-2 related antigens were detected in all of the 30 liver tissues from patients with liver cirrhosis and the expression of TIMP-1 was higher than that of TIMP-2 (P < 0.01). The staining signal of TIMP-1 and TIMP-2 related antigen was mainly located in the cytoplasm of liver cells. There was no signal in nuclear of liver cells. None of the normal liver tissues was positive in expression of TIMP-related antigen. The positive rate was 100% for serum TIMP-1 and 77% for serum TIMP-2. CONCLUSION: Expression of TIMPs at different degrees can be found in liver tissue with cirrhosis. TIMPs are located in cytoplasm of liver cells of patients with cirrhosis. There is a significant correlation between serum TIMPs level and liver TIMPs level. Serum TIMP-1 and TIMP-2 can be used as diagnostic indexes for liver fibrosis in chronic liver diseases.

Adult↗

Hepatitis C virus RNA detection by in situ RT-PCR in formalin-fixed paraffin-embedded liver tissue. Comparison with serum and tissue results.

The purpose of this study was to localize HCV RNA in formalin-fixed paraffin-embedded liver biopsies of 15 patients with chronic hepatitis C using in situ RT-PCR method. The results were compared to serum and tissue extract analysis of HCV RNA. HCV RNA was detected in 80% of the sera tested, in 40% of the corresponding hepatic tissue extract and in 60% of the tissue sections tested by in situ RT-PCR. Compared to the serum positive cases, 67% of the cases were positive with in situ RT-PCR and 41% were positive with tissue extract detection. 50% of the cases in situ RT-PCR positive were also positive with tissue extract detection. These results underlined the complementarity of the different methods of viral detection for the precise diagnosis of hepatitis C.

Formaldehyde↗

THE LYMPHOD TISSUES AND IMMUNE RESPONSES OF NEONATALLY THYMECTOMIZED MICE BEARING THYMUS TISSUE IN MILLIPORE DIFFUSION CHAMBERS.

Neonatally thymectomized mice were implanted intraperitoneally at 7 days of age with Millipore diffusion chambers containing either embryonic or neonatal thymus tissue. Mice which received either empty diffusion chambers or no further treatment following neonatal thymectomy served as controls. In contrast to these controls, most of the mice implanted with thymus-filled chambers gained weight satisfactorily, did not develop a wasting syndrome, and had the capacity to produce serum antibodies in response to sheep erythrocytes and to reject allogeneic skin grafts. Lymphoid follicles were present in the lymph nodes, spleen, and intestinal tract of the implanted mice but most still showed some diminution in the population of lymphocytes in both blood and tissues. Control thymectomized mice had markedly depleted lymphoid tissues and low peripheral blood lymphocyte levels. The tissue recovered after 1 to 2 months from the diffusion chambers showed only epithelial-reticular cells but no lymphoid cells. It is suggested that a humoral factor produced by the thymus epithelial-reticular complex may be responsible for endowing lymphoid cells with immunological competence.

Animals↗

Tissue concentrations of tissue polypeptide antigen (TPA) and prostatic specific antigen (PSA) in 42 patients with prostatic carcinoma.

BACKGROUND: Following development of methods to quantitate biochemical markers in aspiration biopsies we showed that tissue concentration of prostate specific antigen (T-PSA) decreased with increasing malignancy while serum PSA increased. We also found that T-PSA predicts the clinical outcome better than earlier used prognostic markers. METHODS: In order to further study biochemical markers in prostatic cancer a membrane protein, tissue polypeptide antigen (TPA), which is a complex of polypeptide fragments of cytokeratins 8, 18, and 19, was quantitated in 42 patients with newly diagnosed carcinoma of the prostate. The samples had previously been analyzed for T-PSA. RESULTS: Correlation to TGM classification showed that higher malignancy is correlated to lower tissue TPA values. There is a significant positive correlation (r(s) = 0.49, P < 0.01) between T-TPA and T-PSA. Pretreatment values of T-PSA, but not T-TPA, had association to time to progression or time to death. CONCLUSIONS: Increasing prostatic malignancy is correlated to decreasing values of T-TPA. This indicates that the concentrations of membrane and secretory proteins are changed in the same direction in tissue during cancer development. Tissue TPA seem to have no prognostic value in endocrine treatment of prostatic carcinoma.

Aged↗

Development and optimization of tissue preparative methodology for DNA content analysis of soft tissue neoplasms.

Data regarding DNA content parameters in soft tissue sarcoma is limited. Development and optimization of tissue specific preparative techniques for DNA flow cytometry was undertaken prior to routine DNA content analysis of soft tissue neoplasms; 154 soft tissue tumors were studied. Dissociation dependent differences in cellular yields, viabilities, maintenance of DNA aneuploid populations, coefficients of variation, and DNA index supported the need for these developmental studies. Fifty-six of eighty-nine patients had DNA aneuploid soft tissue sarcomas. A relationship between DNA aneuploidy and grade was seen in this series with 38% with low grade, 59% with moderate grade, and 69% with high grade tumors demonstrating DNA aneuploid populations (P < 0.005). The mean S-phase fraction for DNA diploid and aneuploid sarcomas was 7.2% and 13.3%, respectively (P < 0.001). When classified by histologic grade of the primary tumor, a greater percentage of metastatic lesions were DNA aneuploid (4 of 7 grade 2 lesions, and 15 of 16 grade 3 lesions). Decreases in cellular yields and rate of DNA aneuploidy were observed in a subgroup of patients with localized high grade sarcoma treated preoperatively, as compared to patients treated with initial surgery. Prospective correlation of DNA content parameters to prognosis and response to cytotoxic therapy are now possible and are ongoing.

Aneuploidy↗

Tissue factor and thrombomodulin expression on endothelial cell-seeded collagen modules for tissue engineering.

The creation of functional tissue engineering constructs to repair or replace diseased tissues requires a well-formed vasculature network within the construct and the endothelial cells lining that vascular bed must display a nonthrombogenic phenotype. A new approach to tissue engineering involves the assembly of smaller components (modules fabricated at the hundred micron scale) into larger constructs. The modules, collagen gel containing the particular tissue cell of interest, are covered with endothelial cells prior to assembly so that the interconnected channels that are formed are lined with endothelial cells, creating a mimic of a vascular network. Here, we confirmed (using confocal microscopy primarily) that the human umbilical vein endothelial cells, seeded on collagen gel modules without a second embedded cell and without flow, bore the molecular markers of low thrombogenicity. Two days, after seeding on the modules, endothelial cells displayed the typical cobblestone morphology, formed tight cell-cell junctions and covered the whole module surface. Immunofluorescence staining showed that at both 2 days and 7 days after seeding, only a few cells expressed tissue factor while this number was dramatically increased after TNFalpha stimulation. On the other hand, thrombomodulin was expressed by the majority of seeded cells and expression was reduced after TNFalpha stimulation.

Collagen↗

Comparison of localized proton NMR signals of skeletal muscle and fat tissue in vivo: two lipid compartments in muscle tissue.

In vivo 1H NMR spectra of small volumes-of-interest (VOI) were localized in human soleus muscle (8 ml) and compared with volume selective spectra of subcutaneous fat tissue and femoral yellow bone marrow (2 ml). All examinations were performed by the double spin echo (PRESS) localization technique. To provide comparability, spectra of different tissues were recorded using identical sequence timing. Clearly improved resolution of the lipid signals of muscle tissue was obtained using long echo times TE > 200 ms. The spectra of muscle tissue exhibit lipid signals that stem from two compartments with a difference of their resonance frequencies of about 0.2 ppm (Larmor frequency difference 12-13 Hz at 1.5 T). The existence of two fatty acid compartments is supported by measurements of the relaxation times and line shape analysis. Both compartments contain fatty acids or triglycerides with similar composition. Probably one compartment corresponds to fat cells within muscle tissue, the other compartment with lower Larmor frequency is located within muscle cells.

Adipose Tissue↗

Comparisons of brain metabolites observed by HRMAS 1H NMR of intact tissue and solution 1H NMR of tissue extracts in SIV-infected macaques.

The objective of this study was to compare ex vivo proton high-resolution magic angle spinning magnetic resonance spectra of intact tissue with those spectra obtained by solution (1)H NMR of brain extracts of the same sample. Sixteen brain tissue samples from simian immunodeficiency virus-infected rhesus macaques from both frontal cortex and putamen were evaluated by comparing brain metabolite quantities of N-acetylaspartate (NAA), choline-containing compounds (Cho), myo-inositol (MI), creatine (Cr), lactate (Lac), glutamate (Glu) and acetate (Ace). The ratios of the individual NMR peak areas of all metabolites relative to the creatine peak area were calculated. Linear regression analysis revealed significant correlations between measurements using the two methods. The strength of the correlations varied depending on the metabolite studied. We found highly significant correlations for NAA/Cr (r2 = 0.77; p < 0.0001), NAA + Ace/Cr (r2 = 0.73; p < 0.0001) and MI/Cr (r2 = 0.75; p < 0.0001). We observed somewhat less strong correlations for Glu/Cr (r2 = 0.54; p < 0.002) and Lac/Cr (r2 = 0.54; p < 0.002). There was a substantially weaker correlation for Cho/Cr (r2 = 0.32; p = 0.02). When plotting the metabolite ratios obtained by 1H HRMAS NMR of the intact tissue sample on the ordinate vs 1H NMR of the tissue extract on the abscissa, most metabolites exhibited a slope close to unity, and a positive intercept probably due to macromolecular contributions to the MAS spectra. The slope for Cho/Cr was substantially less than unity. Generally, samples from the frontal cortex showed a better correlation between intact and extracted tissue samples than putamen. This is most prominent in the cases of NAA/Cr and Cho/Cr. We conclude that both methods provide substantially the same information for most major brain metabolites, with the exception of the Cho resonance.

Animals↗