Brain tumors or gliomas are fatal cancer species with high recurrence rates due to their strong invasiveness. Therefore, the goal of surgery is complete tumor resection. However, the surgery is difficult to distinguish the border because tumors and blood vessels have the same color tone and shape. The fluorescein sodium is used as a fluorescence contrast agent for boundary separation. When the external light source is irradiated, yellow fluorescence is expressed in the tumor, which helps distinguish between blood vessels and tumor boundaries. But, the fluorescence expression of fluorescence sodium depends on the concentration of fluorescein sodium and such analytical data is insufficient. The unclear fluorescence can obscure the boundaries between blood vessels and tumors. In addition, reduce the efficiency of fluorescence sodium use. This paper proposes a protocol of concentration range for fluorescence expression conditions. Fluorescent expression was observed using a near-infrared (NIR) color camera with corresponding dilution using normal saline in 1 ml microtube. The flunoresence emission density range is 1.00 mM to 0.15 mM. The fluorescence emission begin to 1.00 mM and the 0.15 mM discolor. The discolor is difficult to fluorescence emission condition obserbation. Thus, the maximum density range of the bright fluoresecein is 0.15 mM to 0.30 mM. When the concentration range of fluorescein sodium is analyzed based on the gradient of fluorescence expression and the power measurement, the brightest fluorescence is expected to facilitate the complete resection of the tumor. For the concentration range protocol, setting concentration ranges and analyzing fluorescence expression image according to saturation and brightness to find optimal fluorescence concentration are important. Concentration range protocols for fluorescence expression conditions can be used to find optimal concentrations of substances whose expression pattern varies with concentration ranges. This study is expected to be helpful in the boundary classification and resection of brain tumors and glioma.
To investigate exhausted-medium-induced apoptosis in human corneal epithelial(HCE) cells, this study was performed DNA gel electrophoresis, M30 CytoDEATH staining and FAS-FAS ligand ELISA. SV-40 transfected cells were grown to confluency in culture for 7days. The supernatant was harvested and filtered with $0.22{\mu}m$ filter paper. Fresh HCE cells were exposed to the filtered exhausted medium for 1~2 days. Apoptotic cells were prepared for DNA extraction and run the agarose gel for DNA ladder pattern. M30 CytoDEATH was used a tool for easy and reliable determination of very early apoptosis in HCE cells. The control and exhausted medium were assayed for soluble FAS/FAS ligand protein by ELISA. HCE cells exposed to exhausted medium showed a typical DNA ladder pattern. Sporadic M30 CytoDEATH positive cells were detected among HCE cells exposed to exhausted medium. Soluble FAS/FAS ligand levels were not elevated in the exhausted medium compared to the fresh medium control. This study suggests that possible mechanism of exhausted medium induced apoptosis does not include the FAS-FAS ligand system.
To understand how Bursaphelenchus xylophilus kills pine trees, the differences between the effects of B. xylophilus and B. mucronatus on pine trees are usually compared. In this study, the migration and attacking ability of a non-pathogenic B. mucronatus in Pinus thunbergii were investigated. The distribution of B. mucronatus and the number of dead epithelial cells resulting from inoculation were compared with those of the pathogenic B. xylophilus. Although B. mucronatus is non-pathogenic in pines, its distribution pattern in P. thunbergii was the same as that of B. xylophilus. We therefore concluded that the non-pathogenicity of B. mucronatus could not be attributed to its migration ability. The sparse and sporadic attacking pattern of B. mucronatus was also the same as that of B. xylophilus. However, the number and area of the dead epithelial cells in pine cuttings inoculated with B. mucronatus were smaller than in those cuttings inoculated with B. xylophilus, meaning that the attacking ability of B. mucronatus is weaker than that of B. xylophilus. Therefore, we concluded that the weaker attacking ability of B. mucronatus might be the factor responsible for the non-pathogenicity.
We studied the viability and function of islet with monomethoxy polyethylene glycol (mPEG) grafted onto its membrane. Islets were isolated from rat and were repeatedly reacted with activated mPEG (mw 5000) in order to increase grafting density. The density of grafted PEG on the islet membrane was confirmed by Fluorescein-PEG-NHS. An assessment of islet viability using AO / PI staining method showed that multiple PEGylation did not reduce islet viability. The function of PEG grafted islets was evaluated by measuring released insulin from islets. Insulin secreted from the PEGylated islets for 1 h did not show any significant difference compared to control (non-PEGylated) islets. In addition, PEGylated islets responded in the same pattern as control islets in the perifusion test.
Purpose: The purpose of this study was to examine the corneal topographical changes associated with the wearing of everted silicone hydrogel soft lenses. Methods: The shape and fluorescein pattern of everted silicone hydrogel lenses were investigated. The subject wore the silicone hydrogel everted lenses overnight for 8 hours. Objective refractive error and corneal shape were evaluated at baseline, 1, 2, 3, 5, and 7 days after lens wearing and 1,2,3, and 4 days after discontinuation of lens wear. Results: The Fluorescein pattern of everted silicone hydrogel lenses was similar to the reverse geometry lenses with pressure profile. Objective refractive error(sphere power) and corneal refractive power were decreased and corneal shape had changed during the everted silicone hydrogel lenses wear and recovered during the 4 days of discontinuation. Subject experienced no discomfort associated with the everted silicone hydrogel lenses. Conclusions: It appears that everted silicone hydrogel lenses are capable of inducing significant changes in corneal topography, with overnight wear. Further study must be done to help understand these changes to develop a predictable and effective way of using soft contact lenses for corneal reshaping.
Purpose: The change of alignment between RGP lens and cornea according to the lens design was investigated by comparing the areas of fluorescein pattern in central and peripheral regions analyzed by astigmatic degree and corneal type when spherical and aspherical RGP lenses fitted in alignment. Methods: The fluorescein patterns of 90 eyes (19-30 years, $25.12{\pm}3.52$) having with-the-rule astigmatism were analyzed after spherical and aspherical RGP lenses fitted in alignment. Then, their fluorescent areas in central and peripheral regions were calculated and compared for the quantitative evaluation. Results: The case showing concordant base curve between spherical and aspherical RGP lenses in alignment fitting was 72% however, the possibility to have same base curves between spherical and aspherical RGP lenses in alignment fitting was to be less in the case of symmetric bowtietyped cornea and high astigmatism. The fluorescent area in peripheral region of aspherical RGP lens in alignment fitting was smaller than it of spherical RGP lens. Peripheral fluorescent areas in both RGP lenses decreased according to the increase of astigmatic degree and peripheral area in symmetric bowtie-typed corea was smaller than round-typed cornea's peripheral area. In the case of same astigmatic degree, peripheral fluorescent area of aspherical RGP lens was smaller in both corneal types. Conclusions: The results above suggest the changing degree in the alignment between RGP lens and cornea can be varied according to lens design, corneal astigmatism and corneal type. Thus, the results obtained from the quantitative analysis of the alignment between lens design and cornea may be used as the basic information about the establishment of guidelines for RGP lens fitting, the development of proper lens design, and different tear volume in partial regions.
In order to validate the variable aperture channel model that can deal with the non-uniform How rate in flow domain, migration experiments of conservative tracer were performed in two artificial fractures, a parallel and a wedge-shaped fracture. These different fracture shapes were designed to give different flow pattern. The fractures were made from a transparent acrylic plastic plate and a granite slab with dimensions of 10 $\times$ 61 $\times$ 61 cm. Uranine (Fluorescein sodium salt) was used as a conservative tracer. The volumetric flow rates of uranine feed solution were 30 mL/ hr, giving a mean residence time in the fracture of approximately 24 hours for the parallel fracture and 34 hours for the wedge-shaped fracture. The migration plumes of uranine were photographed to obtain profiles in space and time for movement of a tracer in fractures. The photographed migration plume was greatly affected by the geometric shape of fractures. The variable aperture channel model could have predicted the experimental results for the parallel fracture with a large accuracy. It is expected that the variable aperture channel model would be effective to predict the transport of the contaminant, especially, with the flow rate variation in a fracture.
Purpose: In this study, the effect of lens fitting status on the contact area between spherical/aspherical RGP lens and the cornea having different astigmatic degree and corneal type was investigated for guiding the proper selection of RGP lens. Methods: Spherical and aspherical RGP lenses were applied on ninety eyes $(25.12{\pm}3.52years)$ having with-the-rule astigmatism by different fitting status. Then, their central, mid-peripheral and peripheral areas of fluorescein pattern were calculated and compared for the quantitative evaluation of the contact area between spherical/aspherical RGP lens. Results: The central and peripheral areas with the alignment fitting was significant different based on lens design. However, the central area didn't show any significant difference by lens design and corneal type when fitted in steep or flat. When analyzed by the corneal shape, both lenses with alignment and flat fitting had significant difference in central and peripheral areas. However, the central, mid-peripheral and peripheral areas with steep fitting didn't show the difference by corneal types. When analyzed by the astigmatic degree, the central and peripheral areas with alignment fitting changed proportionally to the increase of corneal astigmatism regardless of corneal shape. With steep and flat fitting, however, the central, mid-peripheral and/or peripheral areas in round- and symmetric bowtie-typed corneas showed the conflicting result when compared to those of alignment fitting when analyzed by the astigmatic degree. Conclusions: In this study, it was confirmed that the contact areas of cornea and RGP lens fitted steep and flat status were largely affected by the corneal type and corneal astigmatism rather than RGP lens fitted in alignment status. Also, this result commonly occurred in both spherical and aspherical RGP lenses.
Alginate microspheres, containing fluorescein isothiocyanate-bovine serum albumin (FITC-BSA) or green fluorescent protein (GFP) were prepared and used as a model drug to develop the oral vaccine delivery system. The alginate microspheres were coated with poly-L-lysine or chitosan. Two methods, w/o-emulsion and spray, were used to prepare alginate microspheres. To optimize preparation conditions, effects of several factors on the particle size and particle morphology of microsphere, and loading efficiency of model antigen were investigated. In both preparation methods, the particle size and the loading efficiency were enhanced when the concentration of sodium alginate increased. In the w/o-emulsion preparation method, as the concentration of Span 80 was increased from 0.5% to 2%, the particle size was decreased, but the loading efficiency was increased. The higher the emulsification speed was, the smaller the particle size and loading efficiency were. The concentration of calcium chloride did not show any effect on the particle size and loading efficiency. In the spray preparation method, the particle size was increased as the nozzle pressure $(from\;1\;kgf/m^2\;to\;3\;kgf/m^2)$ and spray rate was raised. Increasing calcium chloride concentration (<7%) decreased the particle size, in contrast to no effect of calcium chloride concentration on the w/o-emulsion preparation method. Alginate microspheres prepared by two methods were different in the particle size and loading efficiency, the particle size of microspheres prepared by the spray method was about $2-6\;{\mu}m$, larger than that prepared by the w/o emulsion method $(about\;2{\mu}m)$, and the loading efficiency was also higher with spray method. Furthermore, drying process for the microspheres prepared by the spray was simpler and easier, compared with the w/o emulsion preparation. Therefore, the spray method was chosen to prepare alginate microspheres for further experiments. Release pattern of FITC-BSA in alginate microspheres was evaluated in simulated intestinal fluid and PBS (phosphate buffered saline). Dissolution rate of FITC-BSA from alginate/chitosan microsphere was lower than that from alginate microsphere and alginate/poly-L-lysine microsphere. By confocal laser scanning microscope, it was revealed that alginate/FITC-poly-L-lysine microspheres were present in close apposition epithelium of the Peyer's patches of rabbits following inoculation into lumen of intestine, which proved that microspheres could be taken up by Peyer's patch. In conclusion, it is suggested that alginate microsphere prepared by spray method, showing a particle size of & $10\;{\mu}m$ and a high loading efficiency, can be used as a model drug for the development of oral vaccine delivery system.
Kim, Kwang-Dae;Kim, Ki-Hyung;Na, Yong-Jin;Lee, Kyu-Sup
Clinical and Experimental Reproductive Medicine
/
v.26
no.3
/
pp.419-432
/
1999
Objective: To establish the evaluation system of the quality of oocytes on the basis of the incidence of cumulus cells apoptosis, to investigate the relationships beween the incidence of cumulus cells and the outcomes of IVF-ET. Method: Thirth-four cycles undergoing controlled ovarian hyperstimulation for IVF-ET with tubal infertility (23 cycles) or unexplained infertility (11 cycles) were included in this study. Cumulus cell masses surrounding mature oocyte and co-culture of embryos with autologous cumulus cells during IVF-ET process. The incidence of apoptosis in cumulus cells was assessed by apoptosis detection kit fluorescein. The effect of co-culture using cumulus cells and the incidence of cumulus cells apoptosis. Results: The results were as follows: 1. The incidence of apoptosis in cumulus cells markedly increased in patients aged 40 or over, while the fertilization rate was greatly decreased in those age group. 2. Apoptosis in cumulus cells was found in both the fertilized oocytes and unfertilized oocytes, but the incidence of apoptosis was higher in unfertilized oocytes. 3. There is no clear correlation between apoptosis in cumulus cells and the number of oocytes retrieved. However, the incidence of apoptosis was increased when the number of oocytes retrieved was 5 and fewer in comparison with $6{\sim}10$. 4. Embryo grade was significantly affected by the incidence of apoptosis in cumulus cells. 5. Pregnancy rate of IVF-ET per cycle was 29.4%, and the pregnant group had the higher fertilization rate and a significantly lower incidence of apoptosis in cumulus cells compared with the nonpregnant group. 6. When cumulus cells were used as helper cells in the co-culture of the embryo, in vitro activity of cumulus cells based on morphological change and proliferation did not influence the quality of embryo, but was closely associated with the implantation rate and pregnancy rate, which was enhanced when morphological changes and proliferation of cumulus cells was more active. 7. This difference in the outcome of IVF-ET according to in vitro activity of cumulus cells used for co-cultue was not associated with the incidence of apoptosis in cumulus cells; but rather had likely relations with the different secretion pattern of protein, which may be an embryo trophic factor by cumulus cells. Conclusion: These results suggest that the incidence of apoptosis in cumulus cells can be used in predicting oocyte qualities and the outcomes of IVF-ET. And the effect of co-culture largely depends on the in vitro activity of cumulus cells as well.
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