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Biomedical subjects

K Shih

Publications and source records attributed to K Shih.

7 recordsLinked to original sources

Cytokine chemokine expression in contused rat spinal cord.

Spinal cord injury within the first few hours, is complicated by inflammatory mechanisms, including the influx of monocyte/macrophages as well as the activation of resident spinal microglia and astrocytes. Numerous studies have suggested that the initial infiltration of the hematogenous cells may be due to the secretion of cytokines and chemokines in the injured CNS. In order to elucidate which chemotactic factors may be expressed following traumatic spinal cord contusion, the presence of mRNA for a number of cytokines, chemokines and growth factors was examined in contused rat spinal cord by reverse transcriptase-polymerase chain reaction and immunohistochemistry. Spinal injury was accompanied by an increase in glial fibrillary acidic protein mRNA suggesting astrocyte activation and astrogliosis. TNFalpha message levels were upregulated as early as 1 h post injury and returned to baseline levels by 3 days post injury (DPI). By immunocytochemistry, staining for TNFalpha increased at 1 and 3 dpi and was predominantly diffuse in the necrotic tissue. The chemokines IP-10, MCP-1, and MIP-1alpha were also detected in the injured spinal cord. mRNA levels of IP-10 peaked around 6 h post injury and were upregulated up to 7 dpi. MCP-1 mRNA was detected at 1 h post injury and its levels returned to baseline by 14 dpi. An increase in MCP-1 staining was observed from 1 to 7 dpi. The staining was also diffuse in the necrotic tissue and also localized to cells near the site of injury. The presence of aFGF and bFGF was also detected in the injured spinal cord. mRNA for aFGF was detected at 0 time, increased at 6 h post injury, peaked at 3 days, and remained elevated up to 21 days. bFGF mRNA was initially detected at 1 h post injury, increased between 6 h and 3 days, declined thereafter and returned to baseline levels by 21 days.

Animals↗

Quantitative cleaning efficacy evaluation methods with protein and lipid lens soiling models.

We developed a nondestructive, quantitative method for evaluating the cleaning efficacy of rigid gas permeable contact lens care systems. Simulated tears prepared with lysozyme, albumin, and calcium ions were applied to rigid gas permeable lenses as a protein soiling model, and a mixture of grease and unsaturated fat was applied as a lipid soiling model. The ComfortCare system (Pilkington Barnes Hind) and the Boston Cleaner and Boston Conditioning Solution (Polymer Technology) were evaluated for their efficacy in removing protein deposits; water was used as a control. The ComfortCare system and the Boston Advance Cleaner and Boston Advance Conditioning Solution were evaluated for their effectiveness in removing lipid deposits; saline was used as a control. Protein deposit level was detected by a densitometer at the absorbance of 280 nm. The lipid deposit level was evaluated by the absorbance of the fluorescence at 430 nm using the same densitometer with a fluorescence accessory. The two test systems demonstrated a significantly higher cleaning efficacy than the control for both soiling models. The methods provide quantitative measurements and therefore can be analyzed statistically for screening or comparison purposes.

Contact Lens Solutions↗

Antimicrobial efficacy of two commercial RGP contact lens care systems.

We compared the antimicrobial activities of two rigid gas permeable (RGP) lens care systems, a one-step wetting and soaking system (0.005% chlorhexidine gluconate, 0.02% EDTA) and a conditioning solution (0.006% chlorhexidine gluconate, 0.05% EDTA). The formulations were individually inoculated with approximately 10(6) CFU/mL of a pathogenic organism. Both the one-step wetting and soaking system and the conditioning solution demonstrated good antimicrobial efficacy against Pseudomonas aeruginosa, Staphylococcus epidermidis, Serratia marcescens, and Aspergillus fumigatus. However, the one-step wetting and soaking system provides better disinfection against Candida albicans (1.2 log reduction in 4 hours) than the conditioning solution (0.39 log reduction in 4 hours). The one-step system also reduced S. marcescens by 5 logs in 1 hour, whereas the conditioning solution reduced S. marcescens by about 2 logs only in the same time interval. Although both RGP lens care systems use the same active preservative (chlorhexidine digluconate), the variation in disinfecting activities may be attributed to differences in formulation and buffer systems.

Colony Count, Microbial↗

Gentle Touch, a lens material resistant to protein deposition.

We investigated the ability of the Gentle Touch, a non-ionic contact lens material containing 35% netrafilcon A and 65% water, to repel protein deposits. Studies were conducted in vivo (lenses worn by patients) and in vitro (lenses coated by artificial tears). Acuvue soft contact lenses, an ionic lens material containing 42% etafilcon A and 58% water, were used as controls. After 1 to 2 weeks of wear, protein deposits on Gentle Touch lenses averaged 1.5-2.8 micrograms per lens versus 338-444 micrograms per lens for Acuvue. Gentle Touch lenses that were heat-coated with an FDA tear solution recommended for testing protein deposition averaged 6.5 micrograms of protein deposits per lens, whereas almost 100 micrograms of protein was deposited on each Acuvue lens. Lenses treated with an artificial tear solution deposited a non-detectable amount of protein on each Gentle Touch lens versus more than 358 micrograms on each Acuvue lens. These studies demonstrated that Gentle Touch lenses are highly resistant to protein deposits.

Biocompatible Materials↗

Enhancement of liposome-mediated gene transfer to human airway epithelial cells by replication-deficient adenovirus.

We hypothesized that replication-deficient adenovirus (Ad), when complexed with plasmid DNA (pl) and cationic liposomes (L), would enhance liposome-mediated gene transfer in cultured human airway epithelial cells. Pl/L/Ad complexes were formed using charge-charge interactions. A gel electrophoresis retardation assay showed plasmid DNA to be associated with the virus in a high-molecular-weight, low-mobility complex, the diameter of which was 300 to 350 nm. Compared to pl/L alone, pl/L/Ad enhanced luciferase expression on average by 1 log-fold in human airway epithelial cells which express either mutant or wild-type cystic fibrosis transmembrane conductance regulator (CFTR). Transgene expression was sustained at high levels for up to 7 days following transfection with pl/L/Ad. Using a heat-stable alkaline phosphatase reporter gene, we showed that a larger fraction of cells was transfected by pl/L/Ad compared to pl/L. Finally, cells were exposed to Ad for 0 to 24 hours prior to pl/L or exposed to pl/L prior to Ad. We found that enhancement was significantly greater using pl/L/Ad compared to the simultaneous addition of Ad with the pl/L complexes. In addition, when pl/L was added 4 to 24 hours prior to Ad, some enhancement was found, suggesting that plasmid DNA remained in a compartment in the cell for several hours and became available for transcription with the addition of Ad. When Ad was added prior to pl/L, enhancement was found suggesting that the effect of the virus on cell membranes may persist for up to 24 hours. We conclude that the tripartite pl/L/Ad complex significantly enhances liposome-mediated transgene expression for a prolonged period of time in human bronchial epithelial cells.

Adenoviridae↗

Ultraviolet radiation for the sterilization of contact lenses.

Two sources of ultraviolet (UV) radiation with peak wavelengths in the UV-C or UV-B ranges were compared for their ability to sterilize contact lenses infected with Pseudomonas aeruginosa, Streptococcus pneumoniae, Acanthamoeba castellani, Candida albicans, and Aspergillus niger. Also examined was the effect of prolonged UV light exposure on soft and rigid gas permeable (RGP) contact lenses. The UV-C lamp (253.7 nm, 250 mW/cm2 at 1 cm) was germicidal for all organisms within 20 minutes but caused destruction of the soft lens polymers within 6 hours of cumulative exposure. UV-C caused damage to RGP lenses in less than 100 hours. The UV-B lamp (290-310 nm, 500 mW/cm2 at 1 cm) was germicidal for all organisms tested (except Aspergillus) with a 180-minute exposure and caused less severe changes in the soft lens polymers than did the UV-C lamp, although cumulative exposure of 300 hours did substantially weaken the soft lens material. RGP materials were minimally affected by exposure to 300 hours of UV-B. Ultraviolet light is an effective germicidal agent but is injurious to soft lens polymers; its possible utility in the sterilization of RGP lenses and lens cases deserves further study.

Acanthamoeba↗