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Biomedical subjects

B Kelley

Publications and source records attributed to B Kelley.

At least 19 recordsLinked to original sources
PubMed2001

Recombinant human factor VIII-specific affinity ligands selected from phage-displayed combinatorial libraries of protein A.

Factor VIII-specific affibodies were selected from phage displayed libraries constructed by combinatorial mutagenesis of an alpha helical bacterial receptor domain derived from staphylococcal protein A. Bead-immobilized recombinant human factor VIII (rVIII) (80 and 90 kDa chains) protein was used during competitive biopannings in the presence of free 80-kDa chain protein, resulting in the selection of several binders that showed dissociation constants (Kd) in the range 100-200 nM as determined by biosensor analyses. One variant (Z[rVIII:3], 90-kDa chain specific) was further characterized in small-scale affinity chromatography experiments, and showed efficient and selective recovery of biologically active rVIII from Chinese hamster ovary cell supernatant-derived feed stocks. The purity of the enriched rVIII was comparable with rVIII material purified by immunoaffinity chromatography using a 90-kDa chain-specific monoclonal antibody. Interestingly, epitope mapping showed that the monoclonal antibody and the affibody ligand competed for the same or at least overlapping epitopes on rVIII. In addition, the Z[rVIII:3] variant was produced by peptide synthesis with a C-terminal cysteine to enable directed coupling to solid supports. This 59-residue protein was analyzed by circular dichroism and showed a secondary structure content similar to that of the parental Z domain used as scaffold. In biosensor studies, the synthetic affibody was immobilized recruiting the C-terminal cysteine residue, and demonstrated to bind both recombinantly produced and plasma-derived factor VIII. From a secondary library, constructed by re-randomization of relevant positions identified after alignment of the first-generation variants, a panel of affinity-improved second-generation affibodies were selected of which one clone showed a dissociation constant (Kd) for rVIII of 5 nM. Several of these variants also showed higher apparent binding efficiencies towards rVIII when analyzed as immobilized ligands in biosensor experiments. Taken together, the results suggest that affibody ligands produced by bacterial or synthetic routes could be of interest as an alternative to monoclonal antibodies in purification processes or as diagnostic or monitoring tools.

Amino Acid Sequence↗
PubMed2001

Chronic diseases surveillance of St. Regis Mohawk Health Service patients.

A study of the distribution of some chronic diseases among the Akwesasne Mohawks was performed using computerized medical records of the St. Regis Mohawk Health Services Clinic. Prevalence proportions, annual incidence rates, and five-year incidence rates were computed for the period January 1, 1992 to January 1, 1997 for asthma, diabetes mellitus type II, hypothyroidism, and osteoarthritis. Hypothyroidism and diabetes showed higher age-specific prevalence than in the general U.S. population. Osteoarthritis was extremely frequent among people 60 years of age and older, and it may also be elevated in prevalence in relation to the U.S. general population. The incidence and prevalence trends of diabetes type II and osteoarthritis were stationary, but those for asthma and hypothyroidism showed increases over the study period. Morbidity from asthma and acquired hypothyroidism should be monitored in the future and investigated through analytic epidemiologic methods for a possible association with lifestyle and environmental factors.

Adolescent↗
PubMed1998

SCID mice with HIV encephalitis develop behavioral abnormalities.

Severe combined immunodeficient (SCID) mice inoculated intracerebrally (i.c.) with HIV-infected human monocytes develop brain pathology similar to that in humans with HIV encephalitis. This includes HIV-positive macrophages and multinucleated giant cells, astrogliosis, microglial nodules, and neuronal dropout. These xenografts survive about 1 month. To develop a model of chronic HIV encephalitis and to assay the resulting behavioral abnormalities, we reinoculated SCID mice i.c. every 4 weeks for 3 months with either HIV-infected human monocytes (n = 5) or uninfected human macrophages (n = 4) or administered no inoculation (n = 6); these three groups were monitored for behavioral abnormalities. Tests of cognitive function in a Morris water maze 3.5 months after the first inoculation suggested that HIV-infected mice performed poorly compared with controls. Following testing in the water maze on days 4 and 5 of acquisition, motor activity of infected mice was reduced in comparison with that of controls. Retention of goal location when tested 1 week later was impaired in HIV-infected mice compared with controls. Histopathologic analysis of brains revealed significant astrogliosis and strongly suggested higher numbers of major histocompatibility complex (MHC) class II-positive multinucleated macrophages in HIV-infected compared with control mice. Thus, our preliminary studies indicate that SCID mice with HIV encephalitis develop behavioral abnormalities reminiscent of human disease. These behavioral abnormalities are associated with significantly increased astrogliosis, the presence of HIV, and probably multinucleated giant cells. These studies further support the use of this SCID animal model system for studies of the pathogenesis of HIV encephalitis and for drug interventions.

AIDS Dementia Complex↗
PubMed1998

The manufacturing process for recombinant factor IX.

Advances in recombinant DNA manufacturing technology have now made possible the production of a highly purified and active recombinant factor IX (rFIX) product. Recombinant factor IX was developed by (1) stable insertion of the genes for both factor IX and PACE-SOL (a truncated, soluble serine protease needed to enhance the capacity of cells to remove the amino-terminal propeptide from rFIX) into Chinese hamster ovary cells; (2) selection of a cell line that was capable of expressing high amounts of active rFIX while growing in bioreactors containing a completely defined culture medium that does not contain blood or plasma products; and (3) inclusion of four independent chromatography steps, none of which require monoclonal antibodies. Furthermore, rFIX has been extensively tested to demonstrate similarity to plasma-derived factor IX and has been shown to be a consistent, high-purity product. For example, a high-specific-activity product (276+/-23 IU/mg) has been consistently produced throughout 65 consecutive batches from five consecutive manufacturing campaigns. Thus, rFIX offers a consistent and high-purity source of factor IX treatment for patients with hemophilia B.

Animals↗
PubMed1997

Acetabular labrum tear in a 15-year-old male: diagnosis with correlative imaging.

A 15-year-old male with unresolved hip pain after minor trauma is presented. The diagnosis of an acetabular labral tear was made using correlative scintigraphic and magnetic resonance (MR) imaging, obviating the more invasive investigations of arthrography or arthroscopy of the hip. The clinical and radiological features of this lesion are reviewed. Undiagnosed, acetabular labral tears may result in considerable morbidity. Appropriate operative or conservative management of the condition usually provides symptomatic relief.

Accidental Falls↗
PubMed1997

Effects of case management on the nursing context--perceived quality of care, work satisfaction, and control over practice.

PURPOSE: To examine in 1995 the effects of case management on the context of nursing practice-perceived quality of care delivered, work satisfaction, and control over nursing practice-rather than the commonly studied effect on fiscal outcomes. DESIGN: Prospective quasi-experimental. Sampled were nurses on four units at one community hospital where patients with DRG 107 (Coronary Artery Bypass Graft, with no cardiac catheterization) were traditionally hospitalized. METHODS: Registered nurses on these units completed quality of care, satisfaction, and practice-control scales before and 1 year after implementation. FINDINGS: Significant positive differences were found in several aspects of perceived quality of care for both staff nurses and case managers, with specific increases found in nurses' perceived ability to develop relationships with patients, ability to be therapeutic, and support for good care from the institutional structure and administration. A significant decrease in nurses' satisfaction with their pay and other rewards as well as respect from colleagues was found. Case managers were found to have significantly increased perceptions of control over their practice. CONCLUSIONS: Case management was found to have a primarily positive effect on nurses-staff and case managers alike. There were significant increases in several aspects of the nurses' perceptions of the quality of care delivered. Additionally, case managers were more satisfied with the administration, the respect they received, and their pay and rewards in the institution. Most strikingly, case managers perceived themselves to have more control over their practice. Some negative effects of the program were a decrease in satisfaction with the pay and rewards by the staff who were not case managers.

Adult↗
PubMed1996

Intermittent catheterization the right way! (Volume vs. time-directed).

Intermittent catheterization (ICP) is a well-proven effective means of urologic management for spinal cord diseased (SCD) persons who meet the following criteria: adequate low pressure bladder capacity (350-400 cc minimum), adequate hand function, unobstructed urethra and compliant, understanding, continent, cooperative patients. Time-directed (Q4 H-Q6 H), ICP-obtained volumes on twenty-one patients revealed a majority of early, unnecessary as well as some late over-distended bladder catheterizations. The PCI 5000 or "Bladder Manager", a miniaturized ultrasonic bladder volume measuring device developed by Diagnostic Ultrasound of Seattle, was evaluated. It allowed the patients to perform volume-directed ICP which results in less frequent catheterizations and prevents bladder overdistension.

Equipment Design↗
PubMed1980

Hormonal requirements of baby hamster kidney cells in culture.

Baby hamster kidney cells (BHK) were able to proliferate in the complete absence of serum in synthetic medium supplemented with insulin, transferrin, fibroblast growth factor and epidermal growth factor on cell culture dishes coated with fibronectin. Although the addition of individual supplements had little or no effect on cell growth, the combination of the supplements resulted in a significant synergistic effect. The defined medium was also capable of supporting the clonal growth of BHK cells in the absence of serum. Studies on the effect of hormonal supplementation in the presence of low concentrations of fetal calf serum suggest that the function of serum is to supply hormones and growth factors for cell growth.

Animals↗