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Biomedical subjects

S E Wilson

Publications and source records attributed to S E Wilson.

At least 55 records · Page 3Linked to original sources
PubMed2000

Control of directionality in the site-specific recombination system of the Streptomyces phage phiC31.

The genome of the Streptomyces temperate phage phiC31 integrates into the host chromosome via a recombinase belonging to a novel group of phage integrases related to the resolvase/invertase enzymes. Previously, it was demonstrated that, in an in vitro recombination assay, phiC31 integrase catalyses integration (attP/attB recombination) but not excision (attL/attR). The mechanism responsible for this recombination site selectivity was therefore investigated. Purified integrase was shown to bind with similar apparent binding affinities to between 46 bp and 54 bp of DNA at each of the attachment sites, attP, attB, attL and attR. Assays using recombination sites of 50 bp and 51 bp for attP and attB, respectively, showed that these fragments were functional in attP/attB recombination and maintained strict site selectivity, i.e. no recombination between non-permissive sites, such as attP/attP, attB/attL, etc., was observed. Using bandshifts and supershift assays in which permissive and non-permissive combinations of att sites were used in the presence of integrase, only the attP/attB combination could generate supershifts. Recombination products were isolated from the supershifted complexes. It was concluded that these supershifted complexes contained the recombination synapse and that site specificity, and therefore directionality, is determined at the level of stable synapse formation.

Attachment Sites, Microbiological↗
PubMed2000

Glycoprotein IIb/IIIa receptor inhibitor attenuates platelet aggregation induced by thromboxane A2 during in vitro nonpulsatile ventricular assist circulation.

A recent development in antithrombotic research allows the inhibition of platelet aggregation via protection of the glycoprotein IIb/IIIa receptor on the platelet membrane. We hypothesized that a GP IIb/IIIa receptor inhibitor would inhibit thromboxane-induced platelet aggregation during circulation in our in vitro ventricular assist device (VAD) circuit and preserve long-term platelet function. Twenty-one in vitro nonpulsatile centrifugal VAD circuits were simulated for 4 days using 450 ml of fresh human whole blood with or without glycoprotein IIb/IIIa receptor inhibitor (tirofiban). Platelet aggregation and degranulation were measured in whole blood induced by ristocetin, collagen, ADP, and thromboxane A2 (TXA2). The tirofiban-treated group preserved the platelet count and tended to exert these beneficial effects by inhibiting pathologic platelet aggregation induced by TXA2, collagen, and ADP as well as degranulation. Tirofiban may be useful in preserving platelet number and function during clinical VAD use.

Adenosine Diphosphate↗
PubMed2000

The role of beta-lactam antimicrobials as single agents in treatment of intra-abdominal infection.

Broad-spectrum beta-lactam antibiotics have several advantages in the treatment of intra-abdominal infections. These agents are effective against gram-negative rods and anaerobes, reach therapeutic levels rapidly after parenteral administration, and, in the absence of penicillin allergy, generally exhibit low toxicity. The second-generation cephalosporins (e.g., cefoxitin, cefotetan) are used widely in surgical prophylaxis, trauma, and treatment of mild-to-moderate community-acquired infections, but limitations in their spectra and microbial resistance restrict their utility in more serious infections. Extended-spectrum penicillin/beta-lactamase-inhibitor combinations are effective in the treatment of intra-abdominal infections and include enterococci in their spectrum. Gram-negative aerobe resistance has developed to ampicillin/sulbactam. Piperacillin/tazobactam, a ureidopenicillin with increased gram-negative coverage and enhanced antipseudomonal activity, has proved to be effective in clinical trial therapy for intra-abdominal infections. The very broad spectrum carbapenems--imipenem/cilastatin and meropenem--are effective for serious infections or resistant organisms and are often used in the intensive care unit or for nosocomial intra-abdominal infection. These classes of beta-lactams comprise a range of antimicrobials that can be targeted effectively as single agents to both prevention and treatment of intra-abdominal infection.

Abdomen↗
PubMed2000

Incidence and prevention of epithelial growth within the interface after laser in situ keratomileusis.

PURPOSE: To analyze the incidence of epithelial growth within the interface after laser in situ keratomileusis (LASIK) and describe a technique that may prevent its occurrence. METHODS: Results were reviewed for all LASIK procedures performed by two surgeons using identical surgical technique with either the Automated Corneal Shaper or Hansatome microkeratome. After laser ablation, the bed and posterior surface of the flap were irrigated with 0.2 microM filtered basic salt solution, and the surfaces were swept with lint-free sponges to clear the interface of epithelial and other debris. An aspirating lid speculum connected to suction was used to evacuate irrigation fluid and debris from the field. After the flap was smoothed into position, a bandage contact lens was used for the first day after surgery to prevent lifting of the flap edge during blinking. RESULTS: Seven hundred eighty-three eyes of 419 patients had primary LASIK, and 108 eyes had LASIK reoperation. Three eyes developed epithelial growth within the interface, with a minimum of 3 months of follow-up. The interface opacity appeared within 1 month of surgery in all three eyes. Two epithelial nests were small (1.0 and 1.5 mm in diameter, respectively) and did not increase in size on subsequent visits. Neither peripheral epithelial nest affected best-corrected visual acuity or caused any other sequelae. One ingrowth occurred early postoperatively in a case with a donut-shaped flap and was eliminated by transepithelial photorefractive keratectomy (PRK). CONCLUSION: Irrigating, wiping the stromal interface with sponges. aspirating irrigation fluid and debris with a suctioning lid speculum, and use of a bandage contact lens for the first day after surgery during LASIK and LASIK enhancement may markedly reduce the incidence of epithelial growth within the interface.

Acetates↗
PubMed2000

Surgically induced astigmatism after photorefractive keratectomy with the excimer laser.

PURPOSE: To evaluate retrospectively the effect of spherical excimer laser photorefractive keratectomy (PRK) on astigmatism. METHODS: Four hundred seventy consecutive eyes of patients who had PRK for the treatment of myopia without astigmatic keratotomy, PRK reoperation, or other surgical procedures were evaluated in a retrospective clinical study. PRK was performed using the Summit Apex excimer laser with attempted corrections from 1 to 7 diopters (D) of myopia. Preoperative and postoperative astigmatism was determined by manifest refraction refined with a 0.25-D Jackson cross cylinder and evaluated with vector analysis. RESULTS: Eighty-five ( 18%) eyes continued to have a spherical refraction after PRK, 53 (11%) eyes had the same preoperative astigmatism, and 332 (71%) eyes had a change in magnitude of astigmatism > or =0.25 D after spherical PRK. The absolute change in astigmatism magnitude irrespective of axis was +0.4 +/- 0.4 (standard deviation) D at 6 months after PRK. Eyes with change in astigmatism power tended to have higher preoperative myopia and higher preoperative astigmatism. Vector analysis revealed surgically induced astigmatism was 0.68 +/- 0.50 D (range, 0-3.25 D) at 1 month and 0.56 +/- 0.47 D (range, 0-3.1 D) at 12 months after spherical PRK. CONCLUSION: Spherical excimer laser PRK is associated with significant surgically induced astigmatism that is likely related to decentration of the ablation, excimer laser beam irregularities, and variations in wound healing across the ablated zone. Surgically induced astigmatism will complicate attempts to correct astigmatism simultaneously at the time of PRK and suggest that such attempts are likely to be problematic for lower levels of astigmatism.

Adult↗
PubMed2000

One-year results of PRK in low and moderate myopia: fewer than 0.5% of eyes lose two or more lines of vision.

PURPOSE: To retrospectively evaluate the results of myopic photorefractive keratectomy (PRK) for different levels of intended correction, including analysis of loss of best spectacle-corrected visual acuity. METHODS: Four hundred seventy-five consecutive eyes with 1 year of follow-up that had PRK for the correction of 1-7 diopters (D) of myopia by using the Summit SVS Apex excimer laser. Three hundred forty-eight eyes were examined at 1 year. This study was confined to the 236 eyes with 1 year of follow-up that had PRK without astigmatic keratotomy. Eyes also were analyzed according to the range of attempted correction (0-3 D, low; 3.1-6D, moderate; and > or =6.1 D, high moderate). Manifest refraction, uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), surface regularity index (SRI), and surface asymmetry index (SAI) were evaluated for each group. RESULTS: One year after PRK, 91% of all eyes were within 1 diopter and 73% of eyes were within 0.5 diopter of emmetropia. Uncorrected visual acuity was 20/25 or better in 79% and 20/40 or better in 96% of eyes. Two lines of BSCVA were lost in only 0.4% of eyes (one of 236). No eye lost >2 lines of BSCVA, and 30% gained one line. Mean SRI and SAI were increased as compared with preoperative values, but were within the normal range for our patient population (ranges, 0.2-1.0 and 0.1-0.7, respectively). UCVA, BSCVA, and predictability decreased, whereas SAI and SRI increased, with increasing attempted correction. CONCLUSION: PRK effectively reduced myopia in all eyes with 12 months' follow-up. Predictability tended to decrease with increasing attempted correction, even for low to moderate myopia. PRK may induce mild surface asymmetry and irregularity, and these alterations tend to increase with higher attempted correction. Fewer than 0.5% of eyes lost > or =2 lines of best-corrected visual acuity.

Adult↗
PubMed2000

Role of apoptosis in wound healing in the cornea.

Anterior keratocytes undergo apoptosis in response to epithelial injury. This likely represents mechanical activation of systems that vigilantly monitor the corneal epithelium for injury produced by pathogenic viruses. Keratocyte apoptosis is the first change noted after procedures in which the epithelium is injured and may be an initiator of the subsequent wound healing cascade. The working hypothesis is that inhibition of keratocyte apoptosis will limit the subsequent wound healing response. The keratocyte apoptosis response varies with the specific type of epithelial injury induced by a particular refractive surgical procedure, such as photorefractive keratectomy or laser in situ keratomileusis. Continuing efforts aim to identify pharmacological agents that effectively inhibit keratocyte apoptosis without promoting keratocyte necrosis.

Apoptosis↗
PubMed2000

Bowman's layer structure and function: critical or dispensable to corneal function? A hypothesis.

PURPOSE: The purpose of this article is to review available information regarding development, structure, and function of Bowman's layer in the cornea. Disease-related abnormalities of Bowman's layer are described. A hypothesis is advanced to explain the development and maintenance of Bowman's layer. METHODS: Literature review and hypothesis formulation based on previous studies. RESULTS: Information is presented that supports the hypothesis that Bowman's layer forms as a result of cytokine-mediated interactions occurring between corneal epithelial cells and keratocytes that include chemotactic and apoptotic effects on the keratocytes. This hypothesis suggests that Bowman's layer results from such interactions beginning in early development and continuing into adulthood in humans and other animals, such as chickens. CONCLUSIONS: Bowman's layer may be a visible indicator of ongoing stromal-epithelial interactions in the human and have no critical function in corneal physiology. Bowman's layer is commonly destroyed in diseases such as advanced bullous keratopathy where stromal-epithelial interactions may be interrupted. Bowman's-like layers often form in response to epithelium, for example when epithelial plugs extend into the stroma in corneas with radial keratotomy incisions.

Animals↗
PubMed2000

Modulation of TNF-alpha-induced apoptosis in corneal fibroblasts by transcription factor NF-kappaB.

PURPOSE: Previous studies have suggested no role for tumor necrosis factor (TNF)-alpha in the modulation of apoptosis in corneal fibroblasts. However, recent investigations have demonstrated that nuclear factor (NF)-kappaB activation by TNF-alpha mediates negative apoptotic effects that must be blocked to unmask the apoptotic effects of TNF-alpha in vitro. The purpose of this study was to investigate the role of transcription factor NF-kappaB in the suppression of TNF-alpha-induced apoptosis of corneal fibroblasts. METHODS: mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) and RNase protection assay. Proteins were detected by immunocytochemistry and immunoprecipitation with Western blot analysis. Cell death was evaluated by trypan blue exclusion assay in corneal fibroblasts treated with TNF-alpha in presence or absence of the specific inhibitor of NF-kappaB activation, SN50, actinomycin D, or actinomycin D with dexamethasone, ketorolac tromethamine, or diclofenac sodium. Apoptosis was monitored by trypan blue exclusion, colorimetric cell assay, CPP32 activation assay, DNA fragmentation assay, and transmission electron microscopy. NF-KB activation was monitored using electrophoretic gel shift assay. RESULTS: TNF-alpha, TNF receptor (R)I, and TNFRII mRNAs were detected in all three cultured corneal cell types and in ex vivo corneal epithelium using RT-PCR. TNF-alpha mRNA was also detected in ex vivo corneal epithelium, corneal epithelial cells, and stromal fibroblasts with the RNase protection assay. TNF-alpha, TNFRI, and TNFRII proteins were detected by immunocytochemistry in all three major corneal cell types in human corneal tissue. TNF-alpha protein was also detected in ex vivo corneal epithelium, primary corneal epithelial cells, and primary stromal fibroblasts using immunoprecipitation and Western blot analysis. TNF-alpha stimulated corneal fibroblast cell death when NF-kappaB activation was blocked with actinomycin D or SN50. Enhanced cell death was noted with dexamethasone, ketorolac tromethamine, or diclofenac sodium when used in the presence, but not in the absence, of actinomycin D. A gel shift assay revealed induction of NF-KB by TNF-alpha and suppression of induction in the presence of actinomycin D or SN50, but not by the control peptide SN50M. CONCLUSIONS: The TNF-alpha receptor system is expressed in the cornea, and NF-kappaB activation is an important regulator of TNF-alpha-mediated corneal fibroblast apoptosis. Nonsteroidal anti-inflammatory agents or corticosteroids may potentiate corneal fibroblast apoptosis in response to cytokine stimulation.

Antigens, CD↗
PubMed2000

New epidemiology for postoperative nosocomial infections.

Changes in health care delivery systems over the last decade have resulted in a major increase in outpatient surgery and a higher severity of illness for inpatients. We sought to determine the effects of this change on the epidemiology of postoperative surgical infections. Historical data on incidence and epidemiology of infection were obtained from peer-reviewed articles published between 1960 and 1999 (MEDLINE). All nosocomial infections in 5035 patients admitted to a tertiary-care university hospital surgical intensive care unit between January 1994 and December 1997 were prospectively identified and classified as wound, urinary tract, bloodstream, or pneumonia. Incidence of bacterial isolates at each site was also recorded. From these data we determined infection rates per 100 admissions. We also identified all device-related nosocomial infections and calculated infection rates. Comparisons between time periods were made. In the 1960s wound infections constituted the predominant postoperative infection at 46 per cent. This was replaced by urinary tract infection in the 1970s (44%) and 1980s (32%) and closely followed by bloodstream infections (25%). In the 1990s nosocomial pneumonia became the most common postoperative infection, comprising 43 per cent of surgical intensive care unit infections. Analysis of the bacteriology also revealed changing trends with primarily gram-positive organisms in the 1960s followed by an increase in methicillin-resistant Staphylococcus in the 1970 to 1980s, and currently resistant gram-negative bacteria predominate. The incidence of fungal infections has steadily increased. This survey identified a new epidemiology for postoperative surgical infections. Over the last several decades the reported wound infections have been markedly decreased and there is little change in urinary tract infection. Nosocomial pneumonia with resistant gram-negative bacteria now predominates along with increased incidence of fungal infections. Currently, postoperative infections are now more severe, involve critical organs, and require close monitoring of the changing patterns of pathogens.

Ambulatory Surgical Procedures↗
PubMed1999

The complete genome sequence of the Streptomyces temperate phage straight phiC31: evolutionary relationships to other viruses.

The completed genome sequence of the temperate Streptomyces phage straight phiC31 is reported. straight phiC31 contains genes that are related by sequence similarities to several other dsDNA phages infecting many diverse bacterial hosts, including Escherichia, Arthrobacter, Mycobacterium, Rhodobacter, Staphylococcus, Bacillus, Streptococcus, Lactobacillus and Lactococcus. These observations provide further evidence that dsDNA phages from diverse bacterial hosts are related and have had access to a common genetic pool. Analysis of the late genes was particularly informative. The sequences of the head assembly proteins (portal, head protease and major capsid) were conserved between straight phiC31, coliphage HK97, staphylococcal phage straight phiPVL, two Rhodobacter capsulatus prophages and two Mycobacterium tuberculosis prophages. These phages and prophages (where non-defective) from evolutionarily diverse hosts are, therefore, likely to share a common head assembly mechanism i.e. that of HK97. The organisation of the tail genes in straight phiC31 is highly reminiscent of tail regions from other phage genomes. The unusual organisation of the putative lysis genes in straight phiC31 is discussed, and speculations are made as to the roles of some inessential early gene products. Similarities between certain phage gene products and eukaryotic dsDNA virus proteins were noted, in particular, the primase/helicases and the terminases (large subunits). Furthermore, the complete sequence clarifies the overall transcription map of the phage during lytic growth and the positions of elements involved in the maintenance of lysogeny.

Amino Acid Sequence↗
PubMed1999

Patterns of graduating medical student career selections from 1993 to 1998 and their effect on surgery as a career choice.

HYPOTHESIS: Despite successful efforts in California to increase the number of graduating medical students entering primary care residencies, general surgery remains a popular career choice. DESIGN: Retrospective survey of annual graduating medical student residency selections from allopathic medical schools in California between 1993 and 1998. METHODS: Each medical school in California was sent a survey of its graduates' specialty choices from 1993 to 1998. Once quantified, comparisons using t tests, chi2 analyses, and analyses of variance were performed to assess differences in graduating medical student career choices. RESULTS: Surveys were received from 7 of 8 allopathic medical schools in California representing 5154 students (3645 from University of California medical schools). In the study period, the number entering primary care residencies rose from 45.5% in 1993 to 54.3% in 1998. Despite this overall trend, statistically significant increases were seen only for pediatrics between 1993 and 1994. The number of students entering general surgery did not change significantly. Significant decreases were seen in emergency medicine and obstetrics and gynecology between 1997 and 1998 and in anesthesiology between 1994 and 1995 and again between 1995 and 1996. When considering only University of California graduates, the number choosing primary care residencies rose from 46.6% in 1993 to 54.2% in 1998. As before, the number entering general surgery did not change (6.1% in 1993 and 6.3% in 1998). CONCLUSION: Owing to institutional, legislative, and market pressures, more graduates of California medical schools (both public and private institutions) are choosing to become primary care practitioners. A corresponding decrease in medical students entering general surgery has not resulted, which points to an optimistic future and continuing demand for surgeons.

California↗
PubMed1999

Improved survival following massive transfusion in patients who have undergone trauma.

HYPOTHESIS: Survival following massive transfusion in patients who have undergone trauma has improved during the past 10 years. DESIGN: Retrospective cohort study. SETTING: Academic level I trauma center in an urban community. PATIENTS: All patients who underwent trauma and who received greater than 50 U of packed red blood cells or whole blood in the 48 hours following admission to the emergency department. INTERVENTIONS: Data were obtained from blood bank records, the trauma registry, patient medical records, and hospital purchasing records. Patients were divided into 2 groups for comparison (early [1988-1992] and late [1993-1997] periods). MAIN OUTCOME MEASURES: Survival and changes in trauma care provision. RESULTS: Survival following massive transfusion in patients who have undergone trauma has significantly increased during the past 10 years (16% vs 45%, early vs late period, P = .03). Factors associated with poor outcome included male sex, major vascular injury, high Injury Severity Score, severe acidosis, prolonged hypotension, refractory hypothermia, and decreased use of platelet transfusion (all P<.05). In the later period, there was more aggressive correction of coagulopathy, more efficient use of warming measures, decreased operative times for the initial operation, and increased use of component therapy (all P<.05). CONCLUSIONS: Survival following massive transfusion has significantly (P = .03) increased during the past 10 years. Factors that may have contributed to this include more effective and efficient rewarming procedures, improved application of damage control techniques, more aggressive correction of coagulopathy, and improved blood banking procedures.

Adult↗
PubMed1999

Corneal epithelium-specific mouse keratin K12 promoter.

Keratins are structural proteins expressed by epithelial cells. Approximately 30 different keratin proteins have been identified, each with a specific expression pattern in different epithelial cells. The tissue-specific promoter of several keratin genes have been used to direct the expression of transgenes in animals. Keratin K12 and K3 are expressed in differentiated and stratified corneal epithelium, although the relative expression of each appears to vary between species. We targeted the mouse K12 keratin gene in order to develop a tissue-specific promoter that could be utilized to study the functions of genes of interest expressed in the corneal epithelium. Three 5' truncated fragments of the keratin K12 promoter (1.03, 0.71 and 0.25 Kb) showed higher functional and tissue-specific promoter activity in a human corneal epithelial cell line than other cell lines. The 0.25 Kb K12 promoter fragment was also active in cultured rabbit corneal epithelial cells. Thus, increased expression in corneal epithelial cells directed by fragments of the mouse K12 promoter extended across species lines. The paired box homeotic gene 6 (PAX-6), which is involved in controlling eye development, stimulated the activity of keratin K12 promoter.

Animals↗
PubMed1999

Expression of HGF, KGF, EGF and receptor messenger RNAs following corneal epithelial wounding.

Hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), epidermal growth factor (EGF), and their receptors have been associated with homeostasis and wound healing in the cornea. The purpose of this study was to examine the expression of the messenger RNAs for these growth factors and receptors in a wounded series of mouse corneas using in situ hybridization. In situ hybridization was performed with 3H-labeled riboprobes on unwounded corneas and corneas at 30 minutes, 4, 12, 24, 48 and 72 hr, and 7 days after epithelial scrape wounds in Balb/C mice. Qualitative and semi-quantitative analyses were performed. Expression of HGF, KGF and EGF mRNAs in keratocytes in the unwounded cornea was low. EGF mRNA was also expressed in unwounded corneal epithelium. Following wounding, however, these growth factor mRNAs were markedly upregulated in keratocytes. EGF mRNA expression in the epithelium appeared unaffected by wounding. At seven days after wounding and several days following closure of the epithelial defect, HGF mRNA and KGF mRNA were still expressed at higher levels in keratocytes compared with unwounded corneas. No difference in expression of HGF or KGF mRNAs between limbal, peripheral corneal, or central corneal keratocytes was noted in the unwounded cornea, KGF receptor mRNA was prominently expressed throughout the unwounded corneal epithelium. HGF receptor mRNA and EGF receptor mRNAs were expressed at low levels in unwounded cornea epithelium. Following scrape injury, expression of HGF receptor mRNA and KGF receptor mRNA were markedly upregulated in the corneal epithelium, while no significant increase in EGF receptor mRNA expression was noted. These studies suggest a prominent role for HGF and KGF in modulating corneal epithelial wound healing following injury. Less prominent changes in EGF mRNA and EGF receptor mRNA in the corneal epithelium following wounding may suggest that EGF has more of a role in homeostasis in the mouse corneal epithelium.

Analysis of Variance↗