Mouse subcutaneous tissue reaction to calcium hydroxide-based.

Mouse subcutaneous tissue reaction to an embedded calcium hydroxide-based root canal filling material was analyzed histopathologically. After the material was placed within the mouse dorsal subcutaneous tissues, we performed examinations using histopathological, histochemical and immunohistochemical techniques. Two weeks after embedment, the proliferation of granulation tissue had already begun to surround the calcification.Most of the cells observed were macrophages. Likewise, multinucleated giant cells increased significantly. The multinucleated giant cells were observed as two types. In one, the centers of the giant cells were vacuoles, while in the others there were deeply stained calcifications with hematoxylin. Twelve weeks after embedment of the materials, further growth of multinucleated giants cells were sighted. Histochemically, von Kossa-stainpositive granules were observed within the macrophages and multinucleated giant cells as black fine granules. According to the TRAP stained specimens, the multinucleated giant cells especially reacted strongly at 4 weeks. However, the reaction became very weak at 12 weeks. CD68 immunohistochemical staining showed positive reactions in the cytoplasm of the proliferating macrophages and multinucleated giant cells. These results suggest that multinucleated giant cells are present in the surrounding tissues due to implantation of the calcium hydroxidebased root canal filling material, and that the presence of ACP in the cells is due to ingested calcium during active phagocytosis, which would disappear later on

Overview of Cytological Dynamics of Periodontal Ligament Inflammatory Lesions

Cyto-pathological features of the periodontal ligament tissue inflammatory lesions have somehow been carried out but detailed cellular dynamics remain unclear. Therefore, in this review, we overviewed mainly our recent experimental model studies. That is performed using using ordinary ddY mice and BMP bone marrow transplanted mouse model. Regaring the experimental apical inflammatory periodontitis, at four weeks, micro-CT confirmed the presence of a radiolucent image at the apex of the tooth, which was then removed for histological examination. The results showed granulation tissue with fibrosis gradually formed at the periphery of an abscess. Next, if perforation were large, granulation tissue would grow to form periodontal polyp. Results of micro-CT revealed alveolar bone resorption and widening of periodontal ligament. Histopathological examination showed proliferation of fibroblasts with some round cells and blood vessels in the granulation tissue. Double immunofluorescent staining of GFP and Runx2 revealed that both proteins were expressed in spindle-shaped cells. Double immunofluorescent staining of GFP and CD31 revealed that both proteins were expressed in vascular endothelial cells in morphologically distinct vessels. The results suggest that fibroblasts, periodontal ligament fibroblasts and blood vessels in granulation tissue were derived from transplanted-bone marrow cells. Thus, essential growth of granulation tissue in periodontal polyp was caused by the migration of undifferentiated mesenchymal cells derived from bone marrow, which differentiated into fibroblasts and later on differentiated into other cells in response to injury

Histopathology

The aim of this study was to establish a model, which can be used to investigate the response of periodontal tissues to excessive occlusal loading in mice by observing histopathological changes. The experiment was performed on ten 7-week-old ddY male mice. Under general anesthesia by intraperitoneal injectionpentobarbital sodium, a micro-plus-screwpin (head part, 1.7mm in diameter, thickness 0.5mm thickness) was screwed into the pulp cavity of the upper-left-first molar. R_mCT images of the experimental site indicated the occlusal contact position between the upper- and lower-left-first molars during the experimental periods. A micro-plus-screwpin was maintained at a constant position on the occlusal surface throughout the experimental period. Histopathological changes of the periodontal ligament at the furcation lesion of the lower-left-first molar and its surrounding periodontal tissues were observed under a light microscope. The densities of deeply dyeing cells in hematoxylin staining with a round-shaped nucleus were increased in the periodontal ligament, with a peak effect of day 4. Multinucleated giant cells appeared in the central lesion of the periodontal ligament on day 7. The distribution of resorption on the surface of both of the cementum and the alveolar bone was accompanied by multinucleated giant cells, which expanded rapidly from day 7 to day 14. These results showed that histopathological changes of periodontal tissues to excessive occlusal load were observed at the furcation area of molar teeth. The present method confirmed the effectiveness of the experimental model to examine the occlusal trauma on periodontal tissues produced by excessive occlusal load

Matrix-free laser desorption ionization mass spectrometry as a functional tool for the analysis and differentiation of complex phenolic mixtures in propolis: a new approach to quality control

Matrix-free laser desorption ionization (LDI) is a rapid and versatile technique for the ionization of small, UV-light-absorbing molecules. Indeed, many natural products such as polyphenols exhibit inherent LDI properties, potentially facilitating their detection from highly complex samples such as crude extracts. With this in mind, the present work thoroughly evaluated the potential of LDI as an analytical tool for the chemical profiling and differentiation of propolis samples obtained from different global regions. Propolis is a complex bee product containing, among others, significant amounts of phenolic constituents that may show LDI effects. The present work will demonstrate that LDI not only provides reproducible and highly specific fingerprint spectra for each of the tested samples, it further allows their clear differentiation by principal compound analysis (PCA). Contrary to classical analytical approaches such as LC- or GC-MS, LDI does not require time-consuming sample preparation and method optimization procedures. Thus, the technique represents a most interesting analytical tool and potent supplement to classic LC-MS for quality control of herbal pharmaceuticals and dietary supplements. Present results clearly support this approach and further suggest the use of LDI as a versatile tool for the automated analysis of large sample batches on an industrial scale

Quantitative contrast-enhanced ultrasonographic assessment of naturally occurring pancreatitis in dogs

Background: Quantitative contrast‐enhanced ultrasonography (CEUS) can detect pancreatic perfusion changes in experimentally induced canine pancreatitis. However, its usefulness in detecting perfusion changes in naturally occurring pancreatitis is unclear. Hypothesis/Objectives: To determine the feasibility of using CEUS to detect pancreatic and duodenal perfusion changes in naturally occurring canine pancreatitis. Animals: Twenty‐three client‐owned dogs with pancreatitis, 12 healthy control dogs. Methods: Dogs diagnosed with pancreatitis were prospectively included. CEUS of the pancreas and duodenum were performed. Time‐intensity curves were created from regions of interest in the pancreas and duodenum. Five perfusion parameters were obtained for statistical analyses: time to initial up‐slope, peak time (Tp), time to wash‐out (TTW), peak intensity (PI), and area under the curve (AUC). Results: For the pancreas, Tp of the pancreatitis group was prolonged when compared to controls (62 ± 11 seconds versus 39 ± 13 seconds; P < .001). TTW also was prolonged but not significantly (268 ± 69 seconds versus 228 ± 47 seconds; P = .47). PI and AUC were increased when compared to controls (95 ± 15 versus 78 ± 13 MPV; P = .009 and 14,900 ± 3,400 versus 11,000 ± 2,800 MPV*s; P = .013, respectively). For the duodenum, PI and AUC were significantly increased in the pancreatitis group when compared to controls. Conclusions and Clinical Importance: Contrast‐enhanced ultrasonography can detect pancreatic perfusion changes in naturally occurring canine pancreatitis characterized by delayed peak with prolonged hyperechoic enhancement of the pancreas on CEUS. Additionally, duodenal perfusion changes secondary to pancreatitis were observed

Identification of Lipases Involved in PBAN Stimulated Pheromone Production in Bombyx mori Using the DGE and RNAi Approaches

BACKGROUND: Pheromone biosynthesis activating neuropeptide (PBAN) is a neurohormone that regulates sex pheromone synthesis in female moths. Bombyx mori is a model organism that has been used to explore the signal transduction pattern of PBAN, which is mediated by a G-protein coupled receptor (GPCR). Although significant progress has been made in elucidating PBAN-regulated lipolysis that releases the precursor of the sex pheromone, little is known about the molecular components involved in this step. To better elucidate the molecular mechanisms of PBAN-stimulated lipolysis of cytoplasmic lipid droplets (LDs), the associated lipase genes involved in PBAN- regulated sex pheromone biosynthesis were identified using digital gene expression (DGE) and subsequent RNA interference (RNAi). RESULTS: Three DGE libraries were constructed from pheromone glands (PGs) at different developed stages, namely, 72 hours before eclosion (-72 h), new emergence (0 h) and 72 h after eclosion (72 h), to investigate the gene expression profiles during PG development. The DGE evaluated over 5.6 million clean tags in each PG sample and revealed numerous genes that were differentially expressed at these stages. Most importantly, seven lipases were found to be richly expressed during the key stage of sex pheromone synthesis and release (new emergence). RNAi-mediated knockdown confirmed for the first time that four of these seven lipases play important roles in sex pheromone synthesis. CONCLUSION: This study has identified four lipases directly involved in PBAN-stimulated sex pheromone biosynthesis, which improve our understanding of the lipases involved in releasing bombykol precursors from triacylglycerols (TAGs) within the cytoplasmic LDs