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Biomedical subjects

L Kelley

Publications and source records attributed to L Kelley.

At least 19 recordsLinked to original sources
PubMed2001

Preselective gene therapy for Fabry disease.

Fabry disease is a lipid storage disorder resulting from mutations in the gene encoding the enzyme alpha-galactosidase A (alpha-gal A; EC ). We previously have demonstrated long-term alpha-gal A enzyme correction and lipid reduction mediated by therapeutic ex vivo transduction and transplantation of hematopoietic cells in a mouse model of Fabry disease. We now report marked improvement in the efficiency of this gene-therapy approach. For this study we used a novel bicistronic retroviral vector that engineers expression of both the therapeutic alpha-gal A gene and the human IL-2Ralpha chain (huCD25) gene as a selectable marker. Coexpression of huCD25 allowed selective immunoenrichment (preselection) of a variety of transduced human and murine cells, resulting in enhanced intracellular and secreted alpha-gal A enzyme activities. Of particular significance for clinical applicability, mobilized CD34(+) peripheral blood hematopoietic stem/progenitor cells from Fabry patients have low-background huCD25 expression and could be enriched effectively after ex vivo transduction, resulting in increased alpha-gal A activity. We evaluated effects of preselection in the mouse model of Fabry disease. Preselection of transduced Fabry mouse bone marrow cells elevated the level of multilineage gene-corrected hematopoietic cells in the circulation of transplanted animals and improved in vivo enzymatic activity levels in plasma and organs for more than 6 months after both primary and secondary transplantation. These studies demonstrate the potential of using a huCD25-based preselection strategy to enhance the clinical utility of ex vivo hematopoietic stem/progenitor cell gene therapy of Fabry disease and other disorders.

3T3 Cells↗
PubMed1999

Hepatitis B vaccination in pregnancy: factors influencing efficacy.

OBJECTIVE: To determine seroprotective antibody response after hepatitis B vaccination during pregnancy and to assess factors influencing the rate of maternal seroprotection. METHODS: Records of 80 healthy gravidas who elected hepatitis B vaccination during pregnancy, after being identified as hepatitis B surface antigen (HbsAg) and antibody (HbsAb) negative on initial prenatal screen, were analyzed retrospectively. Each gravida was begun on a series of three recombinant hepatitis B vaccines at 0, 1, and 6 months. At 36-40 weeks' gestation, all gravidas were rescreened for seroprotective levels of HbsAb using qualitative enzyme-linked immunosorbent assay analysis. The women were grouped by maternal age (less than 25 years or at least 25 years), smoking history, maternal weight, body mass index (BMI) (less than 30, at least 30, less than 34, or at least 34), number of vaccinations received, race-ethnicity, gestational age at vaccination, and vaccination-to-rescreening interval. Data were compared by t test, chi2 test, or Fisher exact test. Stepwise logistic regression analysis was done. RESULTS: At rescreening, 39 (49%) of the 80 women had seroprotective HbsAb conversion. After two vaccinations, obese women (BMI at least 30) (P = .04), women at least 25 years old (P = .04), and women with smoking histories (P = .005) were significantly less likely to respond to the vaccine. Logistic regression analysis for predicting failure of seroprotective response after two vaccinations showed significantly increased odds for severe obesity with BMI at least 34 (odds ratio [OR] 16.2; 95% confidence interval [CI] 1.7, 154.7), smoking history (OR 7.5; 95% CI 2.0, 27.7), and age at least 25 years (OR 3.9; 95% CI 1.1, 14.4). CONCLUSION: Maternal obesity, advancing age, and smoking have negative influences on the efficacy of hepatitis B vaccination in pregnant women.

Adolescent↗
PubMed1999

Correlation of maternal and fetal hepatitis B antibody titers following maternal vaccination in pregnancy.

The objective of this study was to compare the levels of hepatitis B antibody in maternal and cord blood following maternal vaccination in pregnancy and to determine the level of maternal hepatitis B surface antibody (HbsAb) associated with a seroprotective level in cord blood. Thirty-seven (37) healthy gravidas who were identified as seronegative for hepatitis B surface antigen (HbsAg) and antibody (HbsAb) on initial prenatal assessment and subsequently began a series of three vaccinations in the pregnancy with hepatitis B recombinant DNA vaccine (Engerix-B, SmithKline Beecham) were studied. These mothers represented the first group of gravidas delivering between 6/1/97 and 8/1/97 following the iniatiation of a new protocol of offering hepatitis B vaccination to all patients testing seronegative to HbsAg and HbsAb. All gravidas were given a dose of 20 microg into the deltoid muscle utilizing a 1 1/2-inch needle at a schedule of 0, 1, and 6 months. At the time of delivery specimens were obtained for maternal serum levels and paired cord blood levels of HbsAb. Levels were determined utilizing a quantitative enzyme-linked immunoadsorbent assay (ELISA) analysis (AUSAB-EIA Abbott Lab; Abbott Park, IL). A serum titer of > or =10 mLU/mL was considered seroprotective. Maternal and cord blood groups identified by seroprotection status were then sudivided by number of maternal vaccines received. Data were compared using the Student's t-test and Chi-square or Fisher's exact test. Eighteen gravidas (49%) had seroprotective titers at the time of delivery. Of these, 16 (88%) had seroprotective cord blood levels. All maternal specimens with a HbsAb titer > or =35 mLU/mL were associated with cord blood tiers > or =10 mLU/mL. When maternal titers achieved seroprotective levels of HbsAb, there was no difference in the frequency of cord blood seroprotection comparing groups by number of maternal vaccine doses received. When maternal titers of HbsAb achieve seroprotective levels following vaccination, cord blood seroprotection was achieved in 88% of studied patients. Maternal HbsAb titers > or =35 mLU/mL were associated with cord blood seroprotective levels in all cases.

Enzyme-Linked Immunosorbent Assay↗
PubMed1998

Diagnosis of von Willebrand disease type 2N: a simplified method for measurement of factor VIII binding to von Willebrand factor.

Diagnosis of von Willebrand disease Type 2N (vWD 2N), which mimics hemophilia A and its carrier state, is important for accurate genetic counseling and appropriate therapy. To make testing for the disorder more clinically applicable, we developed a simplified method for measurement of factor VIII (FVIII) binding to von Willebrand factor (vWF) using commercially available reagents and standard clinical assays. FVIII binding to vWF was measured by capture of patient vWF by polyclonal antibodies on cyanogen bromide-activated Sepharose beads, reaction with recombinant FVIII, and assay of unbound FVIII by clotting methods. Unbound vWF was measured in patient plasma after capture by the Laurell method. The ratio of bound FVIII/bound vWF was normal in hemophilia A, vWD Type 1, and vWD Type 3 patients, and abnormal in 5 subjects from two families, all of whom had vWD 2N mutations. Patient 1, with FVIII 8 U/dl, vWF: Ag 61 U/dl, vWF:RC 74 U/dl, and FVIII binding nil, was homozygous for the Arg91 Gln mutation. She was followed during pregnancy and delivered an unaffected heterozygous son. Patient 2 had FVIII 8 U/dl, vWF:Ag 73 U/dl, and vWF:RC 71 U/dl, and very low FVIII binding. She was heterozygous for Arg91Gln, as were her mother and sister; no second vWD 2N mutation was found. Her brother, with FVIII 14 U/dl, vWF:Ag 113 U/dl, and vWF:RC 72 U/dl, has no evidence of vWD 2N. With an X-linked inheritance pattern of bleeding tendency, this family is the first reported with combined hemophilia A and vWD 2N.

Adult↗
PubMed1997

Women's perception of pain and distress during intravenous catheterization and urethral mini-catheterization.

A study was done to prospectively compare the pain and distress of urethral mini-catheterization (MC) with the pain and distress of intravenous (IV) catheterization in women. Ten-centimeter visual analog scales were used on which the left end represented "no pain" or "no distress" and the right "the worst pain imaginable" or "extreme distress." Distress was defined to include embarrassment, anxiety, or fear. A convenience sample of 40 adult women who presented to a university emergency department and were undergoing both procedures as part of their management appraised pain and distress after IV placement and urethral MC. The mean IV pain score was 4.2 cm, whereas the mean MC score was only 2.6 cm (P = .02). The IV and MC distress score means were similar (2.3 v 2.6 cm, P = .55). These results suggest that patients perceive urethral MC as less painful than IV placement. The distress scores of these two procedures did not differ and were low in both instances.

Adolescent↗
PubMed1997

A multicenter, randomized, double-blind, placebo-controlled trial of influenza immunization in multiple sclerosis.

We determined the effect of influenza vaccine in patients with relapsing/remitting MS. Considerable controversy surrounds the question of whether to administer influenza vaccines to MS patients. Prevention of a febrile viral illness is clearly desirable in MS, and previous studies suggest that immunization is safe. Despite this, many clinicians avoid vaccination because they fear precipitating an MS exacerbation. We conducted a multicenter, prospective, randomized, double-blind trial of influenza immunization in patients with relapsing/remitting MS. In the autumn of 1993, 104 patients at five MS centers received either standard influenza vaccine or placebo. Patients were followed for 6 months for evaluation of neurologic status and the occurrence of influenza. Influenza was operationally defined as fever > or = 38 degrees C in the presence of coryza, cough, or sore throat at a time when the disease was present in the community. Attacks were defined in the standard manner, requiring objective change in the examination. Patients were examined at 4 weeks and 6 months after inoculation and were contacted by telephone at 1 week and 3 months. They were also examined at times of possible attacks but not when they were sick with flu-like illness. Three vaccine patients and two placebo patients experienced attacks within 28 days of vaccine (no significant difference). Exacerbation rates in the first month for both groups were equal to or less than expected from published series. The two groups showed no difference in attack rate or disease progression over 6 months. Influenza immunization in MS patients is neither associated with an increased exacerbation rate in the postvaccination period nor a change in disease course over the subsequent 6 months.

Double-Blind Method↗
PubMed1996

Concurrent validity of the Beck Depression Inventory as a measure of depression.

The Beck Depression Inventory was administered, along with measures of anxiety, locus of control, and self-concept, to 211 undergraduates at a major southern institution to test for concurrent validity. Correlations of scores on Beck's scale with other scores provide additional support for Beck's scale as a valid measure of depression.

Adult↗
PubMed1988

Biochemical and genetic characterization of Corynebacterium pseudotuberculosis.

Isolates (n = 94) of Corynebacterium pseudotuberculosis were obtained from sheep, goats, horses, and cattle from various parts of the world. The isolates were characterized biochemically and by restriction endonuclease analysis of DNA. We found near homogeneity in the ability of isolates to ferment carbohydrates and to produce urease. All isolates produced phospholipase D and catalase. The ability of isolates from horses to reduce nitrate, the inability of isolates from sheep and goats to do so, and the correlation of this characteristic with results of restriction endonuclease analyses confirmed the existence of 2 biovars of C pseudotuberculosis. We propose that these biovars be referred to as biovar equi for isolates that reduce nitrate and biovar ovis for isolates that fail to do so.

Animals↗
PubMed1986

Polynucleotide antibodies in connective tissue disease: viral markers or disease mediators?

Antibodies to polynucleotides are seen primarily in systemic lupus erythematosus (SLE), but also occur in a variety of other connective tissue diseases. We looked at the prevalence of antinucleotide antibodies (double- and single-stranded RNA and DNA [dRNA, sRNA, dDNA, and sDNA]) in the sera of patients with SLE (70), rheumatoid arthritis (RA) (31), juvenile rheumatoid arthritis (JRA) (68), osteoarthritis (12), and of 22 patients with a preceding viral illness. In comparison with sera from a control population, elevated mean antibody levels to sRNA were found in the sera of all the patients with connective tissue disease, as well as in the sera of patients with preceding RNA, but not DNA, viral infections. Elevated mean levels of antibodies to dRNA were seen in all groups with the exception of RA. Elevated mean antibody titers to sDNA were not seen in patients with JRA nor were they present in the sera of patients with preceding RNA viral infections. Elevated mean anti-dDNA titers were seen only in sera from patients with SLE. High correlation coefficients between the levels of antibodies to sRNA and dRNA in sera from SLE and RA, and between sDNA and dDNA in sera from SLE suggest cross-reactivity of the antibodies in these diseases. Immunization of an elderly population with influenza (RNA) viral vaccine induced antibodies to sRNA only. These studies further document the prevalence of antipolynucleotide antibodies in the sera of patients with connective tissue diseases.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1978

Cytoplasmic DNA synthesis in rhinovirus type 14-inoculated KB cells.

Rhinovirus type 14 (RV14) incuced a transient statistically significant stimulation in synthesis of DNA which appeared between 0 and 3 h post-inoculation in the cytoplasm of high density monolayer cultures of KB cells. Newly synthesized DNA was measured by incorporation of [3H] thymidine into acid-insoluble DNAase-sensitive material and the cytoplasmic location established by cell fractionation and electron microscope radioautographic methods. A minimum of 10 plaque-forming units per cell of RV14 was required to stimulate DNA synthesis which did not occur above 34.5 degrees C, a temperature optimal for virus replication. Cytoplasmic DNA taken from RV14-infected or control cells could be differentiated from the bulk of cell (nuclear) DNA by several criteria, including: (1) RV14 induction of synthesis; (2) lower buoyant density and greater heterogeneity in CsCl and ethidium bromide/CsCl gradients; and (3) a different kinetic complexity upon reannealing. The Cot 1/2 value of cytoplasmic DNA, calclated as 50--100 from reassociation profiles, was about 10-fold less complex than the Cot 1/2 value of nuclear DNA (800-1000). These data rule out the possibility that cytoplasmic DNA arises by random breakage of nuclear DNA during cell disruption and extraction and are compatible with the hypothesis that inoculation of KB cells with RV14 results in stimulation of synthesis of a specific class of cell DNA which is detected in the cytoplasm.

Cell Line↗