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Biomedical subjects

Roger Beuerman

Publications and source records attributed to Roger Beuerman.

11 recordsLinked to original sources
PubMed2006

Defensins knowledgebase: a manually curated database and information source focused on the defensins family of antimicrobial peptides.

The defensins knowledgebase is a manually curated database and information source focused on the defensin family of antimicrobial peptides. The current version of the database holds a comprehensive collection of over 350 defensin records each containing sequence, structure and activity information. A web-based interface provides access to the information and allows for text-based searching on the data fields. In addition, the website presents information on patents, grants, research laboratories and scientists, clinical studies and commercial entities pertaining to defensins. With the rapidly increasing interest in defensins, we hope that the knowledgebase will prove to be a valuable resource in the field of antimicrobial peptide research. The defensins knowledgebase is available at http://defensins.bii.a-star.edu.sg/.

Anti-Bacterial Agents↗
PubMed2006

An outbreak of Fusarium keratitis associated with contact lens wear in Singapore.

CONTEXT: Fungal keratitis is a potentially blinding condition that is rarely seen with contact lens wear. OBJECTIVE: To describe a nationwide outbreak of fungal keratitis caused by Fusarium species among contact lens wearers in Singapore. DESIGN, SETTING, AND PATIENTS: Nationwide, hospital-based case series. All cases of fungal keratitis among contact lens wearers in all ophthalmology departments in Singapore were reviewed along with the charts of all contact lens wearers with culture-proven fungal keratitis from March 2005 through May 2006. A standardized telephone interview was conducted to obtain additional clinical information. MAIN OUTCOME MEASURE: Diagnosis of Fusarium keratitis associated with contact lens wear. RESULTS: During the study period, 66 patients (68 affected eyes) were diagnosed with Fusarium keratitis associated with contact lens wear; the estimated annual national incidence is 2.35 cases per 10,000 contact lens wearers (95% confidence interval, 0.62-7.22). Patients ranged in age from 13 to 44 years (mean [SD], 27.1 [8.4] years), of which 32 (48.5%) were men. The vast majority (65 patients; 98.5%) wore soft, disposable contact lenses; 62 patients (93.9%) reported using 1 brand of contact lens cleaning solution (ReNu, Bausch & Lomb, Rochester, NY), including 42 patients (63.6%) who recalled using ReNu with MoistureLoc. Most patients (81.8%) reported poor contact lens hygiene practices, including overnight use of daily wear contact lenses (19.7%), and use of contact lenses past the replacement date (43.9%). The final best-corrected visual acuity ranged from 20/20 to 20/80. Five patients (5 eyes; 7.4%) required emergency therapeutic or tectonic corneal transplantation. CONCLUSIONS: A new and evolving epidemic of Fusarium keratitis associated with contact lens wear was found in Singapore. Physicians and eye care practitioners worldwide need to be aware of the likelihood of similar outbreaks emerging among contact lens wearers.

Adolescent↗
PubMed2006

Numerical solution of ocular fluid dynamics in a rabbit eye: parametric effects.

Numerical calculations of the aqueous humor dynamics in the anterior chamber of a rabbit's eye are presented to delineate the basic flow mechanisms. The calculations are based on a geometrical model of the eye, which represents the Trabecular meshwork (TM) as a multilayered porous zone of specified pore sizes and void fraction. The outer surface of the cornea is assumed to be at a fixed temperature (corresponding to the ambient temperature), while the iris surface is assumed to be at the core body temperature. Results are obtained for both the horizontal upward-facing orientation of the eye, and the vertical orientation of the eye. Parameters varied include: the temperature difference between the iris and the cornea to underscore the important role of buoyancy in driving the aqueous humor flow; and, the pupil size reflecting different levels of ambient light. Buoyancy is observed to be the dominant driving mechanism for the convective motion in both orientations of the eye. Variations in the pupil size appear to have little influence on the IOP or flow distribution in view of the dominant role of buoyancy in controlling the flow motion. The study provides distributions of the shear stress and flow patterns and delineates the important role of the eye-orientation on these results.

Animals↗
PubMed2006

Expression and regulation of cornified envelope proteins in human corneal epithelium.

PURPOSE: Stratified squamous epithelial cells assemble a specialized protective barrier structure on their periphery, termed the cornified envelope. The purpose of this study was to evaluate the presence and distribution of cornified envelope precursors in human corneal epithelium, their expression in human corneal epithelial cell cultures, and the effect of ultraviolet radiation (UVB) and transglutaminase (TG) inhibition on their expression. METHODS: Tissue distribution of small proline-rich proteins (SPRRs) and filaggrin and involucrin was studied in human cornea sections by immunofluorescence staining. Primary human corneal epithelial cells (HCECs) from limbal explants were used in cell culture experiments. A single dose of UVB at 20 mJ/cm2 was used to stimulate these cells, in the presence or absence of mono-dansyl cadaverine (MDC), a TG inhibitor. SPRR2 and involucrin protein levels were studied by immunofluorescence staining and Western blot analysis. Gene expression of 12 proteins was investigated by semiquantitative reverse transcription-polymerase chain reaction. RESULTS: In human cornea tissue, SPRR1, SPRR2, filaggrin, and involucrin protein expression were detected in the central and peripheral corneal and limbal epithelium. In HCECs, SPRR2 and involucrin proteins were detected in the cytosolic fraction, and involucrin levels increased after UVB. Both SPRR2 and involucrin levels accumulated in the presence of MDC. Nine genes including involucrin, SPRR (types 1A, 1B, 2A, 2B, and 3), late envelope protein (LEP) 1 and 16, and filaggrin were expressed by HCECs. SPRR 4, loricrin, and LEP 6 transcripts were not detected. UVB downregulated SPRR (2A, 2B) and LEP 1 transcripts. CONCLUSIONS: Various envelope precursors are expressed in human corneal epithelium and in HCECs, acute UVB stress differentially alters their expression in HCECs. The expression of envelope precursors and their rapid modulation by UVB supports the role of these proteins in the regulation of ocular surface stress. TG function may be relevant in the regulation of soluble precursors in UVB-stimulated corneal epithelium.

Adolescent↗
PubMed2006

Transglutaminase participates in UVB-induced cell death pathways in human corneal epithelial cells.

PURPOSE: Ultraviolet light (UVB) is known to cause apoptosis in human corneal epithelial cells. This study evaluates the role of transglutaminase in regulating tumor necrosis factor (TNF) receptor clustering as well as caspase activation in UVB-induced apoptosis in human corneal epithelial cells. METHODS: A human corneal epithelial cell line was used. A single dose of UVB (20 mJ/cm2) was used as a stimulus. Cell viability and cell death were investigated by MTT, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL), and caspase-3 assays. Immunofluorescent staining was used to investigate TNF receptor-I clustering at various time intervals after UVB. Short interfering RNA was used to knock down transglutaminase-2 expression. Fluorescein-cadaverine uptake was used to assess transglutaminase activity. A noncovalent peptide delivery system was used to transfect guinea pig liver transglutaminase into corneal epithelial cells. RESULTS: UVB increased transglutaminase activity, reduced cell viability, and increased TUNEL staining. UVB or TNF-alpha promoted TNF-receptor-I clustering, a process inhibited by the transglutaminase inhibitor, mono-dansyl cadaverine. UVB also increased activated caspase-3, in a manner suppressible by mono-dansyl cadaverine. Intracellular delivery of exogenous transglutaminase markedly increase caspase-3 activation compared with the vehicle control. CONCLUSIONS: Transglutaminase enzymatic activity is involved in corneal epithelial cell death after UVB and appears to participate in two steps regulating this process, clustering of TNF receptor-I and caspase-3 activation.

Apoptosis↗
PubMed2006

Human limbal progenitor cell characteristics are maintained in tissue culture.

INTRODUCTION: To determine the differentiation of human limbal epithelial cells in tissue culture. MATERIALS AND METHODS: Epithelial cells from the human limbus (n = 29) were isolated and cultured in supplemental hormonal epithelial medium (SHEM) in the presence of mitomycin C-treated 3T3 feeder layer. Confluent cells were airlifted to form multiple layers. The expression of cytokeratin 3 (K3), cytokeratin 12 (K12), involucrin, connexin 43 (Cx43), proliferation cell nuclear antigen (PCNA) and p63 was studied in normal and airlifted cells by immunohistochemistry. Expression levels of K3 and K12 mRNA were examined by real-time polymerase chain reaction (PCR). RESULTS: The colony-forming efficiency of primary cultured (P0) cells was about 19.35 +/- 6.46% (mean +/- SD, n = 7). Real-time PCR analysis showed that the transcription level of K3 and K12 in cultured cells was lower than in freshly isolated limbal cells or cells from central cornea (P <0.01). Few cells were positive for K3 in P0 or P1 cells [(1.99 +/- 1.27)% (n = 7, P0) and (3.96 +/- 1.35)% (n = 4, P1), P = 0.046]. More cells at all levels were found to stain positive for PCNA and p63 as compared to K3, K12 and involucrin. After air-lifting, cell sheets of 3 to 5 epithelial cell layers formed. Involucrin showed positive staining in suprabasal layers of the cell sheets while connexin 43 was only observed in the basal layer. Staining of K3 remained sparse. CONCLUSIONS: Human limbal cells isolated from cadaveric tissues were able to proliferate in vitro and exhibited a phenotype with characteristics similar to that of the limbal stem or progenitor cells.

Cell Differentiation↗
PubMed2004

The in vitro and in vivo proliferative capacity of serum-free cultivated human conjunctival epithelial cells.

PURPOSE: To investigate the in vitro and in vivo proliferative capacity of human conjunctival epithelial cells cultured in serum-free media, and to compare this with current methods that utilize serum-containing media and 3T3 feeder layers. METHODS: Human conjunctival epithelial cells were cultivated in serum-free media alone, serum-free media with a 3T3 feeder layer, and serum-containing media with a 3T3 feeder layer. The areas of outgrowth, colony-forming efficiencies and number of population doublings were compared. The in vivo proliferative potential was assessed by analyzing the number of cells generated by the implantation of cultured cells into athymic mice. Cultured cells were evaluated for the expression of cytokeratins K3, K4, K12, K19, as well as the gel-forming goblet cell mucin, MUC5AC. RESULTS: Cells cultivated in serum-free media, serum-free media and feeder cells, and serum-containing media and feeder cells achieved colony-forming efficiencies of 14.5 +/- 4.1%, 10.1 +/- 3.1%, and 20.4 +/- 6.7%, respectively, and number of population doublings of 24.8 +/- 4.3, 14.8 +/- 3.6, and 30.0 +/- 5.0, respectively. Nine-day old athymic mice conjunctival cysts derived from serum-free cultures comprised 1.29 x 10(6) +/- 0.46 x 10(6) cells, while cysts derived from serum-containing cultures comprised 1.30 x 10(6) +/- 0.53 x 10(6) cells. The degree of epithelial stratification was similar in both conditions. Serum-free cultivated conjunctival cells retained their in vivo characteristics and expressed K4, K19 and MUC5AC. The presence of MUC5AC mRNA in these cells was confirmed by RT-PCR. CONCLUSIONS: Conjunctival epithelial cells propagated in serum-free media demonstrated a similar in vivo proliferative capability, as compared to serum-containing media with 3T3 feeder cells. This has important clinical implications, as the serum-free ex vivo expansion of cells for clinical transplantation overcomes the problems associated with the use of animal serum and cells.

3T3 Cells↗
PubMed2003

Early gene expression in the rat cortex after experimental traumatic brain injury and hypotension.

To determine c-fos and egr-1 mRNA levels in the rat cortex after traumatic brain injury (TBI) and TBI plus hemorrhagic hypotension (HH). Sprague-Dawley rats were subjected to TBI. HH was induced by removing 5 ml of blood from the jugular vein for a duration of 20 min. Cortical tissue was harvested at 0 and 120 min, and mRNA levels of the immediate early genes c-fos, egr-1 were obtained. Rats that sustained TBI alone had increased c-fos and egr-1 mRNA expression at 120 min that was significant. Additionally, c-fos mRNA levels after TBI alone were significantly higher than c-fos mRNA levels after TBI+HH at 120 min. Levels of egr-1 mRNA after TBI alone at 120 min were not significantly higher than egr-1 mRNA levels after TBI+HH at 120 min.

Animals↗
PubMed2002

Response of GAP-43 and p75 in human neuromas over time after traumatic injury.

OBJECTIVE: GAP-43 and p75 are proteins that promote growth cone and neurite formation, elongation, and arborization in regenerating nerve axons. The objectives of this study were to determine whether GAP-43 and the low-affinity nerve growth factor receptor p75 are elevated in traumatic neuromas and whether there is a correlation between the relative amount of GAP-43 or p75 and demographic characteristics such as time elapsed between injury and repair. METHODS: Traumatic neuromas from 21 randomly selected patients were studied, and the charts were reviewed. Specimens were collected at the time of nerve resection and grafting. Immunohistochemical analysis was performed on each sample and normal human nerve with antibodies to GAP-43 and p75. Western blot and computerized gel analyses were performed. RESULTS: All neuroma specimens harvested within 13 months of injury exhibited markedly elevated GAP-43 levels compared with normal nerve. Specimens harvested at 14 months or more after injury showed precipitously lower GAP-43 levels, similar to or less than those of normal nerve. The correlation between the amount of intra-axonal GAP-43 and postinjury time interval was statistically significant, P = 0.0038. High GAP-43 levels were also correlated with transection injury, high postoperative sensory grade, and pain. p75 levels were elevated, without consistent variation in our population. CONCLUSION: These preliminary data suggest that the expression of intra-axonal GAP-43 may vary over time after injury, remaining elevated for approximately the first year, then decreasing abruptly to normal or subnormal levels. These results correlate with clinical experience, indicating that peripheral nerves should be repaired relatively early if repair is indicated.

Adolescent↗
PubMedDate not supplied

Formation of collagen-glycosaminoglycan blended nanofibrous scaffolds and their biological properties.

The development of blended collagen and glycosaminoglycan (GAG) scaffolds can potentially be used in many soft tissue engineering applications since the scaffolds mimic the structure and biological function of native extracellular matrix (ECM). In this study, we were able to obtain novel nanofibrous collagen-GAG scaffolds by electrospinning collagen blended with chondroitin sulfate (CS), a widely used GAG, in a mixed solvent of trifluoroethanol and water. The electrospun collagen-GAG scaffold with 4% CS (COLL-CS-04) exhibited a uniform fiber structure with nanoscale diameters. A second collagen-GAG scaffold with 10% CS consisted of smaller diameter fibers but exhibited a broader diameter distribution due to the different solution properties in comparison with COLL-CS-04. After cross-linking with glutaraldehyde vapor, the collagen-GAG scaffolds became more biostable and were resistant to collagenase degradation. This is evidently a more favorable environment allowing increased proliferation of rabbit conjunctiva fibroblast on the scaffolds. Incorporation of CS into collagen nanofibers without cross-linking did not increase the biostability but still promoted cell growth. The potential of applying the nanoscale collagen-GAG scaffold in tissue engineering is significant since the nanodimension fibers made of natural ECM mimic closely the native ECM found in the human body. The high surface area characteristic of this scaffold may maximize cell-ECM interaction and promote tissue regeneration faster than other conventional scaffolds.

Animals↗