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Biomedical subjects

K A Walter

Publications and source records attributed to K A Walter.

At least 19 recordsLinked to original sources
PubMed2001

The adverse effect of perioperative brimonidine tartrate 0.2% on flap adherence and enhancement rates in laser in situ keratomileusis patients.

PURPOSE: To investigate the effect of perioperative brimonidine tartrate 0.2% on postoperative flap adherence and enhancement rates in patients undergoing laser in situ keratomileusis (LASIK). DESIGN: Noncomparative interventional case series. PARTICIPANTS: Records of 279 eyes of 144 consecutive patients who underwent LASIK between June 17, 1999, and March 1, 2000, were retrospectively reviewed. METHODS: From September 28, 1999, to November 4, 1999, 39 eyes of 20 consecutive patients had a single drop of brimonidine tartrate 0.2% instilled into their operative eye(s) before LASIK. This group was interposed in time between two nonbrimonidine groups, one before the use of brimonidine, "prior nonbrimonidine group" (107 eyes of 56 patients), and one after the use of brimonidine, "post nonbrimonidine group" (133 eyes of 68 patients). All 279 eyes in the study were examined after surgery to identify flap folds and dislocations. Enhancement rates and visual outcomes for the brimonidine and nonbrimonidine groups were also documented. MAIN OUTCOME MEASURES: Comparison of the incidence of abnormal flap appearance and enhancement rates between the brimonidine and the nonbrimonidine groups. RESULTS: In the brimonidine treated group, 6 of 39 eyes (15.4%) had an abnormal flap appearance after LASIK, although no flap folds or dislocations were noted in the nonbrimonidine groups. The difference in incidence of abnormal flap appearance between the brimonidine group and the prior and post nonbrimonidine groups was statistically significant (P = 0.0008 and P = 0.0003, respectively). When the prior and post nonbrimonidine groups were combined and compared with the brimonidine group, the difference in flap adherence was highly statistically significant (P = 0.00001). The enhancement rate was significantly higher in the brimonidine group at 36% compared with 14% in the prior nonbrimonidine group (P = 0.01) and 9% in the post nonbrimonidine group (P = 0.0004). The mean 3-month postoperative spherical equivalent was significantly greater at -0.88 diopters (D) in the brimonidine group compared with -0.24 D (P = 0.0008) and -0.15 D (P = 0.0001) in the prior nonbrimonidine and post nonbrimonidine groups, respectively. CONCLUSIONS: In this study, when perioperative topical brimonidine was used in LASIK patients, an increase in the incidence of abnormal flap adherence and an increase in enhancement rates was observed.

Adrenergic alpha-Agonists↗
PubMed2000

Scatter factor/hepatocyte growth factor protects against cytotoxic death in human glioblastoma via phosphatidylinositol 3-kinase- and AKT-dependent pathways.

We have shown recently that the multifunctional growth factor, scatter factor/hepatocyte growth factor (SF/HGF), and its receptor c-met enhance the malignancy of human glioblastoma through an autocrine stimulatory loop (R. Abounader et al., J. Natl. Cancer Inst., 91: 1548-1556, 1999). This report examines the effects of SF/HGF:c-met signaling on human glioma cell responses to DNA-damaging agents. Pretreating U373 human glioblastoma cells with recombinant SF/HGF partially abrogated their cytotoxic responses to gamma irradiation, cisplatin, camptothecin, Adriamycin, and Taxol in vitro. This cytoprotective effect of SF/HGF occurred at least in part through an inhibition of apoptosis, as evidenced by diminished terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling index and reduced DNA laddering. Anti-c-met U1/ribozyme gene transfer inhibited the ability of SF/HGF to protect against single-strand DNA breakage, DNA fragmentation, and glioblastoma cell death caused by DNA-damaging agents, demonstrating a requirement for c-met receptor function. Phosphorylation of the cell survival-promoting kinase Akt (protein kinase B) resulted from SF/HGF treatment of U373 cells, and both Akt phosphorylation and cell survival induced by SF/HGF were inhibited by phosphatidylinositol 3-kinase inhibitors but not by inhibitors of mitogen-activated protein kinase kinase or protein kinase C. Cytoprotection by SF/HGF in vitro was also inhibited by transient expression of dominant-negative Akt. Transgenic SF/HGF expression by intracranial 9L gliosarcomas reduced tumor cell sensitivity to gamma irradiation, confirming the cytoprotective effect of SF/HGF in vivo. These findings demonstrate that c-met receptor activation by SF/HGF protects certain glioblastoma cells from DNA-damaging agents by activating phosphoinositol 3-kinase-dependent and Akt-dependent antiapoptotic pathways.

Animals↗
PubMed2000

5-Fluorouracil for the treatment of intraepithelial neoplasia of the conjunctiva and cornea.

OBJECTIVE: To evaluate the efficacy of pulse dosing of topical 5-fluorouracil (5-FU) in the treatment of conjunctival and corneal intraepithelial neoplasia. DESIGN: Prospective, noncomparative case series. PARTICIPANTS: Seven patients with histologic evidence of intraepithelial neoplasia were identified by conjunctival biopsy or tumor excision. METHODS: Seven patients with a minimum of 7 months of follow-up were treated with pulsed dosing of 1% 5-FU. Topical 1% 5-FU was administered four times daily for 2 to 4 days for each cycle. The number of initial treatment cycles was two to six, with the time between cycles being 30 to 45 days. MAIN OUTCOME MEASURES: The presence or absence of clinically evident intraepithelial neoplasia was evaluated after each treatment interval. Patients were also monitored for adverse reactions to the use of topical 5-FU. RESULTS: Four patients remain disease free with a mean follow-up of 18.5 months (range, 7-36 months) with no additional treatment after the initial treatment cycles (mean, 3.75 cycles; range, 2-5 cycles). Three patients had recurrence of disease after the initial treatment cycles. Two patients were treated with additional cycles for recurrent disease (six cycles in one patient and five cycles in the other patient) and are free of disease at 20 and 21 months after treatment, respectively. One patient had persistent disease despite treatment with topical 5-FU and was treated with topical mitomycin C with resolution of the disease without recurrence for 16.5 months. No adverse reactions to pulse dose treatment with topical 5-FU were noted. CONCLUSIONS: Pulsed dosing with 1% topical 5-FU for the treatment of conjunctival and corneal intraepithelial neoplasia, alone or as an adjunct to excision of bulky disease, is a well-tolerated and effective method of treatment.

Administration, Topical↗
PubMed1998

Crystal structure of ErmC', an rRNA methyltransferase which mediates antibiotic resistance in bacteria.

The prevalent mechanism of bacterial resistance to erythromycin and other antibiotics of the macrolide-lincosamide-streptogramin B group (MLS) is methylation of the 23S rRNA component of the 50S subunit in bacterial ribosomes. This sequence-specific methylation is catalyzed by the Erm group of methyltransferases (MTases). They are found in several strains of pathogenic bacteria, and ErmC is the most studied member of this class. The crystal structure of ErmC' (a naturally occurring variant of ErmC) from Bacillus subtilis has been determined at 3.0 A resolution by multiple anomalous diffraction phasing methods. The structure consists of a conserved alpha/beta amino-terminal domain which binds the cofactor S-adenosyl-l-methionine (SAM), followed by a smaller, alpha-helical RNA-recognition domain. The beta-sheet structure of the SAM-binding domain is well-conserved between the DNA, RNA, and small-molecule MTases. However, the C-terminal nucleic acid binding domain differs from the DNA-binding domains of other MTases and is unlike any previously reported RNA-recognition fold. A large, positively charged, concave surface is found at the interface of the N- and C-terminal domains and is proposed to form part of the protein-RNA interaction surface. ErmC' exhibits the conserved structural motifs previously found in the SAM-binding domain of other methyltransferases. A model of SAM bound to ErmC' is presented which is consistent with the motif conservation among MTases.

Amino Acid Sequence↗
PubMed1998

Domain structure analysis of elongation factor-3 from Saccharomyces cerevisiae by limited proteolysis and differential scanning calorimetry.

Elongation-factor-3 (EF-3) is an essential factor of the fungal protein synthesis machinery. In this communication the structure of EF-3 from Saccharomyces cerevisiae is characterized by differential scanning calorimetry (DSC), ultracentrifugation, and limited tryptic digestion. DSC shows a major transition at a relatively low temperature of 39 degrees C, and a minor transition at 58 degrees C. Ultracentrifugation shows that EF-3 is a monomer; thus, these transitions could not reflect the unfolding or dissociation of a multimeric structure. EF-3 forms small aggregates, however, when incubated at room temperature for an extended period of time. Limited proteolysis of EF-3 with trypsin produced the first cleavage at the N-side of Gln775, generating a 90-kDa N-terminal fragment and a 33-kDa C-terminal fragment. The N-terminal fragment slowly undergoes further digestion generating two major bands, one at approximately 75 kDa and the other at approximately 55 kDa. The latter was unusually resistant to further tryptic digestion. The 33-kDa C-terminal fragment was highly sensitive to tryptic digestion. A 30-min tryptic digest showed that the N-terminal 60% of EF-3 was relatively inaccessible to trypsin, whereas the C-terminal 40% was readily digested. These results suggest a tight structure of the N-terminus, which may give rise to the 58 degrees C transition, and a loose structure of the C-terminus, giving rise to the 39 degrees C transition. Three potentially functional domains of the protein were relatively resistant to proteolysis: the supposed S5-homologous domain (Lys102-Ile368), the N-terminal ATP-binding cassette (Gly463-Lys622), and the aminoacyl-tRNA-synthase homologous domain (Glu820-Gly865). Both the basal and ribosome-stimulated ATPase activities were inactivated by trypsin, but the ribosome-stimulated activity was inactivated faster.

Amino Acid Sequence↗
PubMed1998

Retrospective analysis of a novel method of transscleral suture fixation for posterior-chamber intraocular lens implantation in the absence of capsular support.

PURPOSE: To determine the safety and efficacy of an alternative method for transscleral fixation of a secondary posterior-chamber intraocular lens (pcIOL) during penetrating keratoplasty. METHODS: Eighty-nine eyes that underwent secondary pcIOL implantation by using a modified transscleral suture-fixation technique during penetrating keratoplasty were retrospectively evaluated. The surgical technique used suture fixation to the surface of the sclera 5 mm posterior to the limbus, with the knot buried beneath Tenon's capsule and conjunctiva. Patient records were reviewed for postoperative complications, including suture erosion, pcIOL subluxation, vitreous hemorrhage, and retinal detachment. Mean follow-up was 24.4 months, with a range of 4-68 months. RESULTS: All eyes had successful fixation of their pcIOL immediately after surgery. Three (3.3%) eyes had graft failure. Six (6.7%) of 89 eyes showed evidence of suture erosion or partial exposure. Postoperative suture breakage occurred in two (2.2%) eyes. Posterior-segment complications included retinal detachment in one (1.1%) eye, vitreous hemorrhage in one (1.1%) eye, and limited choroidal hemorrhage in two (2.2%) eyes. Median visual acuity at 1-year follow-up was 20/70 (range, 20/25 to light perception). CONCLUSION: This transscleral fixation technique provides a straightforward alternative to previously described techniques. Suture erosion, IOL dislocation, and posterior-segment complications occurred at relatively low rates compared with other pcIOL implantation techniques.

Adult↗
PubMed1997

NMR-based discovery of lead inhibitors that block DNA binding of the human papillomavirus E2 protein.

The E2 protein is required for the replication of human papillomaviruses (HPVs), which are responsible for anogenital warts and cervical carcinomas. Using an NMR-based screen, we tested compounds for binding to the DNA-binding domain of the HPV-E2 protein. Three classes of compounds were identified which bound to two distinct sites on the protein. Biphenyl and biphenyl ether compounds containing a carboxylic acid bind to a site near the DNA recognition helix and inhibit the binding of E2 to DNA. Benzophenone-containing compounds which lack a carboxylic acid group bind to the beta-barrel formed by the dimer interface and exhibit negligible effects on the binding of E2 to DNA. Structure-activity relationships from the biphenyl and biphenyl ether compounds were combined to produce a compound [5-(3'-(3",5"-dichlorophenoxy)-phenyl)-2,4-pentadienoic acid] with an IC50 value of approximately 10 microM. This compound represents a useful lead for the development of antiviral agents that interfere with HPV replication and further illustrates the usefulness of the SAR by NMR method in the drug discovery process.

Antiviral Agents↗
PubMed1997

A new subarachnoid hemorrhage grading system based on the Glasgow Coma Scale: a comparison with the Hunt and Hess and World Federation of Neurological Surgeons Scales in a clinical series.

OBJECTIVE: Although the Hunt and Hess Scale (HHS) and World Federation of Neurological Surgeons Scale (WFNSS) are the most widely used subarachnoid hemorrhage (SAH) grading systems, neither system has achieved universal acceptance. We propose a simplified grading system based entirely on the Glasgow Coma Scale (GCS), which compresses the 15-point GCS into five grades that are comparable with those of the HHS and WFNSS. We refer to this system as the GCS grading system and present a direct comparison with the HHS and WFNSS for predictive value regarding patient outcome and interrater reliability. METHODS: We reviewed 291 consecutive patients with aneurysms treated at our institution between January 1992 and January 1996 and compared the admission grades from the GCS, WFNSS, and HHS with outcome measures at discharge from hospitalization. The Glasgow Outcome score was used as the major outcome measure to evaluate the predictive value of the three scales. Mortality and length of stay (LOS) were also evaluated as outcome measures. The predictive value of each scale was tested with an ordinal logistic regression model for Glasgow Outcome score, a logistic regression model for mortality data, and a linear regression model for LOS. RESULTS: Using the logistic regression model, the GCS was the best predictor of discharge Glasgow Outcome score, with an odds ratio of 2.585 (P = 0.0001), compared with 2.311 (P = 0.0001) for the WFNSS and 2.262 (P = 0.0001) for the HHS. Using mortality data in the logistic model, the HHS was the best predictor, with an odds ratio of 3.391 (P = 0.0001), compared with 2.859 (P = 0.0001) for the GCS and 2.560 (P = 0.0001) for the WFNSS. Each of the three scales had a high predictive value for LOS, using a linear model. We discuss, however, the problematic nature of LOS as an outcome measure for SAH. Interrater reliability for each scale was evaluated using kappa statistics, based on 15 additional patients evaluated prospectively, and showed that the GCS grade also had the greatest interrater reliability, with a kappa of 0.46 (P = 0.0002), compared with 0.41 (P = 0.0005) for the HHS and 0.27 (P = 0.027) for the WFNSS. CONCLUSION: We conclude that the GCS grade has equal or greater predictive value regarding outcome after SAH than do the currently used grading systems and that it has greater reproducibility across observers. Broader familiarity with the GCS among medical and paramedical personnel may further enhance the usefulness of the GCS grade over the HHS and WFNSS in providing a standardized, universally accepted grading system for SAH.

Aneurysm, Ruptured↗
PubMed1997

Aneurysmal subarachnoid hemorrhage: prognostic features and outcomes.

The prognostic features and outcomes associated with aneurysmal subarachnoid hemorrhage (SAH) are reviewed. In the first section, the epidemiology of SAH is discussed with emphasis on prevalence, incidence, risk factors, heredity, activity, and seasonal variability. In the second section, the presentation, diagnosis, and treatment of patients with aneurysmal SAH is briefly reviewed. In the third section, the prognostic features associated with aneurysmal SAH are discussed with emphasis on neurologic condition and SAH grading scales, patient's age, aneurysm size and location, repeat hemorrhage, vasospasm, systemic disease, hypertensive response, computed tomograph features, hydrocephalus, timing of surgery, and expertise of the aneurysm center. Also in the third section, the prognostic features associated with unruptured aneurysms are discussed with emphasis on the actuarial risk of rupture, aneurysm size and location, and multiplicity of lesions. In the fourth and final section, the outcomes of aneurysmal SAH over the past 60 yrs are reviewed.

Aneurysm, Ruptured↗
PubMed1996

Initial characterization of autoprocessing and active-center mutants of CMV proteinase.

Human cytomegalovirus (CMV) encodes a unique serine proteinase that is required in the maturation of the viral capsid. The CMV proteinase can undergo autocatalytic activation and is subject to proteolytic self-inactivation. Mutant enzyme forms were prepared to eliminate the initial autoprocessing site and thus form an active single-chain protein for structure-function studies. Two mutants of CMV proteinase were cloned and expressed in Escherichia coli. The A143V mutant was a conservative substitution at the first internal cleavage site. The S132A mutant modified one of the triad of residues responsible for catalytic activity. Through the use of computer-controlled high-cell-density fermentations the mutant proteins were expressed in E. coli at approximately 170 mg/L as both soluble (approximately 40% of total) and inclusion-body forms (approximately 60% of total). The soluble enzyme was purified by standard methods; inclusion-body protein was isolated by standard methods after refolding and solubilization in guanidine or urea. Sedimentation equilibrium and sedimentation velocity analyses reveal that the enzyme undergoes concentration-dependent aggregation. It exhibits a monomer <==> dimer equilibrium (Kd = 1 microM) at low concentrations and remains dimeric at high concentrations (28 mg/ml). Differential scanning calorimetry data for protein thermal unfolding fit best to a non-two-state model with two components (Tm = 52.3 and 55.3 degrees C) which subsequently aggregate upon unfolding. Analysis of the short-UV circular dichroism spectra of protein forms resulting from expression as soluble molecules (not refolded) reveals that the two mutants have very similar secondary structures which comprise a mixed structural motif of 20% alpha-helix, 26% beta-sheet, and 53% random coil. Though soluble and active (A143V mutant only), CD analysis revealed that protein refolded from inclusion bodies did not exhibit spectra identical to that of protein expressed only in soluble form.

Amino Acid Sequence↗
PubMed1996

Corneal endothelial deposits in patients with cytomegalovirus retinitis.

PURPOSE: We studied six patients with human immunodeficiency virus (HIV) who had cytomegalovirus retinitis and abnormal endothelial deposits in at least one eye, to characterize their corneal endothelial deposits. METHODS: The corneas of the six patients were examined by slit-lamp biomicroscopy and specular microscopy with morphometric analysis. The eyes of one patient with endothelial changes were obtained post mortem for histopathologic and ultrastructural examination. RESULTS: There were multiple diffuse, fine refractile, stellate-shaped deposits on the corneal endothelium in all affected eyes. The deposits were best seen with retroillumination. Two of six patients examined with specular microscopy showed severe abnormalities, which included marked areas of polymegathism and decreased endothelial cell counts. Examination of one eye obtained post mortem disclosed chains of dendritic macrophages and fibrin adherent to the apical surface of the corneal endothelium. There was no evidence of direct infection of the corneal endothelium by cytomegalovirus. CONCLUSIONS: Deposits on the corneal endothelium in patients with cytomegalovirus retinitis most likely result from an anterior uveitis. A preponderance of macrophages observed by histopathologic examination may be related to the inability of the immunodeficient patient to mount a normal T-cell response.

AIDS-Related Opportunistic Infections↗
PubMed1995

Substrate requirements for ErmC' methyltransferase activity.

ErmC' is a methyltransferase that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics by catalyzing the methylation of 23S rRNA at a specific adenine residue (A-2085 in Bacillus subtilis; A-2058 in Escherichia coli). The gene for ErmC' was cloned and expressed to a high level in E. coli, and the protein was purified to virtual homogeneity. Studies of substrate requirements of ErmC' have shown that a 262-nucleotide RNA fragment within domain V of B. subtilis 23S rRNA can be utilized efficiently as a substrate for methylation at A-2085. Kinetic studies of the monomethylation reaction showed that the apparent Km of this 262-nucleotide RNA oligonucleotide was 26-fold greater than the value determined for full-size and domain V 23S rRNA. In addition, the Vmax for this fragment also rose sevenfold. A model of RNA-ErmC' interaction involving multiple binding sites is proposed from the kinetic data presented.

Bacillus subtilis↗
PubMed1995

Intratumoral chemotherapy.

In an effort to improve survival from malignant gliomas, investigators have used intratumoral chemotherapy protocols to deliver high doses of tumoricidal agents directly to the brain. Theoretically, these infusions bypass the blood-brain barrier, minimize systemic drug levels and the side effects of chemotherapy, and achieve prolonged elevations of intracerebral chemotherapeutic agents relative to those obtainable by systemic administration. Almost all major classes of chemotherapeutic agents have been examined as possible intratumoral therapies via delivery approaches ranging from simple intratumoral injections to implantable computer-driven constant infusion pumps and biodegradable polymer matrices. In this review, we summarize the major clinical trials and experimental investigations underlying the development of intratumoral chemotherapy as a treatment for gliomas.

Animals↗
PubMed1994

Interstitial taxol delivered from a biodegradable polymer implant against experimental malignant glioma.

Taxol is a novel antitumor agent with demonstrated efficacy against ovarian, breast, and non-small cell lung cancers in Phase II clinical trials, but which has been shown not to cross the blood-brain barrier. To adapt taxol as a therapy for brain tumors, we have incorporated it into a biodegradable polyanhydride matrix for intracranial implantation and evaluated this formulation in a rat model of malignant glioma. Fischer 344 rats bearing intracranial 9L glioma tumors were treated with 10 mg poly[bis(p-carboxyphenoxy)propane-sebacic acid] (20:80) copolymer discs, containing 20-40% taxol by weight, 5 days after tumor implantation. The taxol-loaded polymers doubled (38 days, 40% taxol loading, P < 0.02) to tripled (61.5 days, 20% taxol loading, P < 0.001) the median survival of rats bearing tumor relative to control rats (19.5 days). Drug loadings of 20-40% taxol by weight released intact taxol for up to 1000 h in vitro. In rats followed up to 30 days postimplant, the polymer maintained a taxol concentration of 75-125 ng taxol/mg brain tissue (100-150 microM taxol) within a 1-3-mm radius of the disc. At points more distant from the disc (up to 8 mm away, the size limit of the rat brain), the polymer maintained a taxol concentration of greater than 4 ng taxol/mg brain tissue (5 microM). We conclude that taxol shows promise as a therapy for malignant glioma when delivered interstitially from a biodegradable polymer.

Animals↗
PubMed1994

Cytotoxicity of taxol in vitro against human and rat malignant brain tumors.

Taxol is a novel antitumor alkaloid that has shown clinical activity against several tumors, including ovarian and breast carcinoma and melanoma. To evaluate taxol's potential as a therapy for malignant brain tumors, we measured the sensitivity of four human (U87, U373, H80, and D324) and two rat (9L, F98) brain-tumor cell lines to taxol. The cells were exposed to taxol in vitro using a clonogenic assay. Log cell kill (LD90) occurred at concentrations of 42 (9L), 25 (F98), 19 (H80), 7.2 (U373), 9.1 (U87), and 3.9 nM (D324) when cells were continuously exposed to taxol for 6-8 days. The human cell lines were uniformly more sensitive to taxol than were the rat lines. The duration of exposure had a significant effect on taxol's cytotoxicity. When cells were exposed to taxol for 1 h the LD90 increased to 890 nM for the 9L rat line and 280 nM for the human U373 line. On the basis of these results, we conclude that taxol has significant potency in vitro against malignant brain tumors and that the activity occurs at concentrations of taxol that have previously been shown to be effective for several tumors against which the drug is currently being evaluated clinically.

Animals↗