Objective: Diabetes mellitus (DM) is known to cause many systemic complications as well as male infertility. Astaxanthin (ASTX) is a powerful antioxidant that is involved in a variety of biologically active processes, including those with anti-diabetes effects. The present study investigates the effect of ASTX on the spermatozoa function in streptozotocin (STZ)-induced diabetic rats. Methods: We divided 30 adult rats into three groups (10 rats per group), with a control group that received corn oil mixed with chow. DM was induced by intra-peritoneal injection of STZ. Eight weeks after the STZ injection, half of the diabetic animals were used as diabetic controls, and the rest were treated with ASTX for 56 days. Then the parameters and chromatin integrity of the epididymal sperm were analyzed using chromomycin A3, toluidine blue (TB), and acridine orange (AO) staining. Results: The count, viability, and motility of the epididymal sperm were decreased significantly in the STZ group in comparison with the control group (count and viability, p<0.001; motility, p<0.01). ASTX increased normal morphology and viable spermatozoa compared to the STZ group (morphology, p=0.001; viability, p<0.05). The percentage of abnormal chromatins in TB and AO staining was higher in the STZ group compared to the control group (p<0.001). The mean percentage of TB and AO positive spermatozoa in STZ rats was significantly lower in the STZ+ASTX group (TB, p=0.001; AO, p<0.05). Conclusion: This study observed that in vivo ASTX treatment partially attenuates some detrimental effect of diabetes. Conversely, ASTX improved sperm viability, normal morphology, and DNA integrity.
This study was performed to elucidate the distribution and correlation of immunoglobulin G subclasses with the degree of inflammation in the experimentally induced rat pulp and periapical pathoses. The pulp exposures were made in 108 mandibular 1st molars of 54 rats and the teeth were left open to the oral environment The animals were sacrified at 3, 7, 15, 30, 60 and 90 days after pulp exposure, and examined microscopically and radiographically Seventy one specimens were routinely sectioned at the thickness of 4 - $6{\mu}$ and stained with Hematoxylin - eosin for histologic examination, with toluidine blue for mast cells, and with the primary antibodies against rat IgG subclasses by using the Avidin - Biotin complex method. The following results were obtained: 1. As the degree of inflammation of rat pulp and periapeces intensified, the number of IgG subclass containing cells per unit area, especially IgG2a and IgG2c, decresased. 2. The IgG2c cells were most predominantly found in the lesions with slight inflammation, IgG1 cells in mild or severe inflammation, and IgG2a cells in moderate inflammation. 3. IgG subclass containg cells were more predominantly observed in the periapical granuloma than periapical abscess or cyst(p<0.01). 4. IgG2a containing cells were predominant in pulp inflammation, IgG1 containing cells in periapical granuloma, IgG2a cells and IgG1 cells in periapical abscess, and IgG2a cells were significantly predominant in periapical cyst. 5. The number of IgG subclass containing cells and mast cells in periapical tissue decreased with time lapse after pulp exposure. And correlation index between mast cells and IgG1, IgG2a, IgG2b was stastically high.
Objective: Preservation of the periodontal ligament (PDL) is vital to the success of tooth autotransplantation (TAT). Increased PDL volumes and facilitated tooth extraction have been observed upon orthodontic preloading. However, it is unclear whether any changes occur in the expressions of bone biomolecules in the increased PDL volumes. This study aimed to determine the expressions of runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), receptor activator of nuclear factor kappa-B ligand (RANKL), and osteoprotegerin (OPG) in PDL upon preloading. Methods: Seventy-two premolars from 18 patients were randomly assigned to experimental groups that received a leveling force for 1, 2, or 4 weeks or to a control unloaded group. Following extraction, PDL volumes from 32 premolars of eight patients (21.0 ± 3.8 years) were evaluated using toluidine blue staining. The expressions of the biomolecules in the PDL from 40 premolars of ten patients (21.4 ± 4.0 years) were analyzed via immunoblotting. Results: The median percentage of stained PDL was significantly higher at 2 and 4 weeks after preloading than in the unloaded condition (p < 0.05). The median RUNX2 and ALP expression levels were significantly higher at 2 and 4 weeks after preloading than in the unloaded condition (p < 0.05), whereas the median RANKL/OPG ratios were significantly higher at 1 and 4 weeks after preloading (p < 0.05). Conclusions: Orthodontic preloading for 4 weeks enhances PDL volumes as well as the expressions of RUNX2, ALP and the RANKL/OPG ratio in the PDL, suggesting this loading period is suitable for successful TAT.
Kim So-Jung;Hwang Eui-Hwan;Lee Sang-Rae;Hong Jung-Pyo
Journal of Korean Academy of Oral and Maxillofacial Radiology
/
v.26
no.2
/
pp.33-44
/
1996
This study was for comparing healing patterns and effects between with odontogenic and nonodontogenic tissues on the defected mandible. Experimental bone defects that measured 3 mm in diameter were created on the mandibular body of guinea pig by removal of bone with the use of trephine burs and bone defects were grafted with Biogran(Orthovita Co., U.S.A.) and covered with Dura Mata(Pfrimmer-Viggo GmbH Co., Germany). Guinea pigs were serially terminated by fours on the 3 days, the 1 week, the 2 weeks, the 3 weeks, the 4 weeks, and the 5 weeks after experiment, and the mandibular body was removed and fixed with 10% neutral formalin. They were decalcified and embedded in paraffin as using the usual methods. The specimen sectioned and stained with hematoxylin and eosin and toluidine blue. They were observed with a light microscope and a polarizing microscope. The obtained results were as follows : 1. Defected bone was healed fast from the odontogenic tissues in early stage of the experiment. 2. The arrangement of the bone matrix was relatively regular in the bone from the nonodontogenic tissues, but irregular in the bone from the odontogenic tissues. 3. Compact bone has started to be resorbed and changed to the pattern of matrix bone tissue from 3 weeks after experiment.
6 beagle dogs aged over one and half years and weighed 14 to 16 Kg were utilized in this study, Horizontal furcation defects were induced around 2nd, 3rd, and 4th premolars bilaterally, PDGF-BB in conjunction with EGF and PDGF-BB only were applied in the right and left premolars respectively. 2 animals were sacrificed at 4weeks, 8 weeks, and 12 weeks, after regenerative surgery respectively. Semi-thin sections using glass-knife were stained with toluidine blue for light microscopic study. The results were as follows: 1. At 4 weeks after regenerative surgery, bone formation in the PDGF-BB-applied site was thriving, but bone formation in the PDGF-BB-and-EGF-applied site was depressed. 2. Bony ankylosis was surely shown along the whole exposed root surface applied with PDGF-BB, but it was shown at the root surface near the base of the bone defect where was applied with PDGF-BB in conjunction with EGF. 3. Active bone formation was made from 8 weeks after regenerative surgery in the PDGF-BB- and-EGF-applied site. 4. Bone maturity as well as speed of bone formation in the PDGF-BB-applied site was superior to those in the PDGF-BB-and-EGF-applied site throughout the whole experimental period. Within the above results, PDGF-BB had the strong capability to form the new bone and EGF was not able to prevent the bony ankylosis thoroughly. However, EGF may have the possibility to prevent the bony ankylosis through the suppression of bone formation.
Objective: To evaluate sperm nuclear DNA fragmentation and chromatin structure after 18 hours' incubation at room temperature. Methods: Twenty-eight male partners who participating IVF treatment were prospectively included in this study. Ejaculated sperm count and motility were assessed. The sperm was then immediately processed by the conventional swim-up method. After utilization of some of the sample for routine clinical use, the remainder of each of the samples was divided into two aliquots. One aliquot was immediately assessed for sperm nuclear DNA fragmentation (TUNEL assay) and chromatin structure (toluidine blue [TB] staining). The other aliquot was incubated at room temperature for 18 hours and then assessed by two methods. Only dark-TB sperms were considered as having abnormal chromatin structure. Data before and after extended incubation were compared using a paired Student's $t$-test. Results: Before and after extended culture, nuclear DNA fragmentation assessed by TUNEL was $4.9{\pm}4.7%$ and $7.0{\pm}6.4%$, respectively ($p$=0.008). The proportion of abnormal chromatin structure (dark-TB sperm) was $8.2{\pm}5.6%$ and $10.3{\pm}6.5%$ ($p$ <0.001), before and after incubation, respectively. Conclusion: After 18 hours' incubation at room temperature, sperm nuclear DNA and chromatin structure were significantly affected. The IVF practitioner should bear this information in mind when performing delayed insemination, especially for $in$$vitro$ maturation cycles.
Polyelectrolyte titration, which was called colloid titration is based on the stoichiometric reaction between oppositely charged polyelectrolytes, This can be used, for instance, to determine the charge density of a cationic polyelectrolyte, using an anionic polyelectrolyte of known charge density, such as potassium polyvinyl sulfate (PPVS). The technique requires a suitable method of end-point detection and there are several possibilities. In this work, two methods have been investigated: visual titrimetry based on the color change of a cationic dye (o-toluidine blue, o-Tb) and spectrophotometry based on the absorbance change corresponding to the color change of the same dye. These have been applied to several cationic polyelectrolytes with different charge density and molecular weight. In all cases, the cationic charge was due to quaternary nitrogen groups. In the case of cationic dye, it was shown that the sharpness depends on the charge density of cationic polyelectrolyte. With the polyelectrolytes of lower charge density, the binding to PPVS is weaker and binding of the dye to PPVS can occur before all of the polyelectrolyte charge has been neutralized. However, by carrying out titrations at several polyelectrolyte concentrations, good linear relationships were found, from which reliable charge density values could be derived. Effects of pH and ionic strength were also briefly investigated. For cationic polyelectrolytes (copolymers of acrylamide and dimethylaminoethy] acrylate), there was some loss of charge at higher pH values, probably as a result of hydrolysis. Increasing ionic strength causes a less distinct color change of o-Tb, as a result of weaker electrostatic interactions.
The endodontic-periodontic combined lesions have been difficult to get correct diagnosis and predictable treatment. This study was to make the experimental endodontic-periodontic combined defects in dogs for the study of the periodontal regeneration and to evaluate the efficacy of the enamel matrix protein and e-PTFE membrane in the experimental endodontic-periodontic combined defects. 5 mongrel dogs were used. The pulp chambers were opened and the plaque was inserted into the chambers to induce the periapical lesions on the mandibular second, third and fourth premolars of the dogs. 1 month later, the root canal treatments were done with gutta perch a and ZOE sealer. On the day of surgery, the periapical defects were standardized by trephine bur. The buccal dehiscence defects were made by the dental bur and bone chisels. The apicoectomy with retrofilling was done. The prepared roots were randomly selected for test and control groups. In the experimental groups, the enamel matrix derivative and e-PTFE membrane were used. Nothing was placed on the control group. Fluroscent labelling was used to evaluate the bone formation. After 4 and 12 weeks, the dogs were sacrificed and undecalcified sections were prepared and stained with toluidine blue. Those histologic sections were examined by fluorescent microscopy and light microscopy. The results were as follows. 1. In the control group, new bone was formed in the periapical defects and scarcely in the buccal dehiscence defects. New cementum was not detected at 4 and 12 weeks. 2. In the experimental groups, new bone, new cementum and periodontal ligament were found in the periapical and buccal dehiscence defects. The relative amount and the quality of the new bone, new cementum and periodontal ligament tissue that had formed on the experimental groups were superior to those of the control group. 3. The current observation implicated that e-PTFE membrane and enamel matrix protein could be the effective tools for the guided tissue regeneration of the endo-perio combined defects.
Primary gastric lymphoma (PGL) is derived from mucosa-associated lymphoid tissue (MALT) and it differs from nodal lymphoma in histologic features and biologic behavior. Recent studies have showed that Helicobacter pylori (H.pylori ) infection is closely related to the development of low grade gastric lymphoma, and eradication of the infection induces regression of the tumor. H. pylori infection is known to be important to the development of gastric MALT lymphoma. The aim of this study was to elucidate the histopathological behavior of PGL according to the concept of MALT and to compare the predictive value of tests frequently used for diagnosis of H. pylori. The histological features of gastric lymphoma arising from MALT are the replacement of glands by uniform dense infiltration of centrocyte-like cells in the lamina propria and lymphoidepithelial lesion. H. pylori-associated histologic changes of neutrophilic infiltration, lymphoid follicle or aggregates formation and intestinal metaplasia, and H. pylori immunoreactivity were analyzed. Detection of H. pylori in chronic active gastritis and peptic ulcer suggests a possible role of H. pylori in the pathogenesis. Giemsa, Toluidine blue and Long H&E stains were used in H. pylori detection. Histopathological examination of gastric biopsy specimens revealed lymphoepithelial lesions pathognomonic of MALT lymphoma, and immunohistochemical staining for CD20 was diffusely positive. CD3 was positive in reactive T cells. PAX-5 was negative except the follicle. Bcl-2, cytokeratin, Ki-67, and c-myc were positive. The findings may indicate a predictable transition of low grade to high grade, and c-myc may be used as a valuable marker before molecular pathology diagnosis.
Bacterial cellulose (BC) is generated from citrus gel by Gluconacetobacter hansenii TL-2C. BC has good properties such as high-burst pressure, high-water contact and the ultrafine highly nanofibrous structure of mimic natural extracellular matrix (ECM) for tissue engineering. In this study, acrylic acid (AAc) was grafted onto BC surfaces under aqueous conditions using gamma-ray irradiation, and then immobilized gelatin onto AAc-g-BC. The characterization of scaffolds was performed by scanning electron microscopy (SEM), attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), toluidine blue O (TBO) assay. Morphology of gelatin and AAc incorporation onto BC nanofibers did not changed. Our study suggests that gelatin-immobilized BC nanofibers scaffold has a potentiality to fabricate 3D nanofibrous scaffolds for tissue engineering.
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