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Biomedical subjects

D Frazier

Publications and source records attributed to D Frazier.

At least 19 recordsLinked to original sources
PubMed2001

Immunoglobulin G3 from polyclonal human immunodeficiency virus (HIV) immune globulin is more potent than other subclasses in neutralizing HIV type 1.

Passive antibody prophylaxis against human immunodeficiency virus type 1 (HIV-1) has been accomplished in primates, suggesting that this strategy may prove useful in humans. While antibody specificity is crucial for neutralization, other antibody characteristics, such as subclass, have not been explored. Our objective was to compare the efficiencies of immunoglobulin G (IgG) subclasses from polyclonal human HIV immune globulin (HIVIG) in the neutralization of HIV-1 strains differing in coreceptor tropism. IgG1, IgG2, and IgG3 were enriched from HIVIG by using protein A-Sepharose. All three subclasses bound major HIV-1 proteins, as shown by Western blot assay and enzyme-linked immunosorbent assay. In HIV-1 fusion assays using X4, R5, or X4R5 envelope-expressing effector cells, IgG3 more efficiently blocked fusion. In neutralization assays with cell-free viruses using X4 (LAI, IIIB), R5 (BaL), and X4R5 (DH123), a similar hierarchy of neutralization was found: IgG3 > IgG1 > IgG2. IgG3 has a longer, more flexible hinge region than the other subclasses. To test whether this is important, IgG1 and IgG3 were digested with pepsin to generate F(ab')(2) fragments or with papain to generate Fab fragments. IgG3 F(ab')(2) fragments were still more efficient in neutralization than F(ab')(2) of IgG1. However, Fab fragments of IgG3 and IgG1 demonstrated equivalent neutralization capacities and the IgG3 advantage was lost. These results suggest that the IgG3 hinge region confers enhanced HIV-neutralizing ability. Enrichment and stabilization of IgG3 may therefore lead to improved HIVIG preparations. The results of this study have implications for the improvement of passive immunization with polyclonal or monoclonal antibodies and suggest that HIV-1 vaccines which induce high-titer IgG3 responses could be advantageous.

Antibodies, Viral↗
PubMed2000

Nonmetallic fixation in elective maxillofacial surgery.

Resorbable fixation technology offers several benefits, including easily cut and shaped plates, strong and predictable resorption qualities, and improved patient acceptance and expectations. Moreover, resorbable fixation implants can be completely reabsorbed into the body, eliminating the need for subsequent removal. This article describes the use of this innovative technology in orthognathic surgery, including preoperative and postoperative patient needs, intraoperative patient care, and potential complications.

Absorbable Implants↗
PubMed1999

Phase I and pharmacokinetic analysis of high-dose tamoxifen and chemotherapy in normal and tumor-bearing dogs.

PURPOSE: To determine whether tamoxifen plasma concentrations capable of blocking P-glycoprotein (Pgp) in vitro can be safely achieved in dogs and whether doxorubicin pharmacokinetic alterations occur when tamoxifen is coadministered. METHODS: Tamoxifen dose escalation studies were conducted in 7 normal dogs and in 19 tumor-bearing dogs receiving full-dose chemotherapy. Plasma tamoxifen and serum doxorubicin disposition were analyzed for putative drug interactions. RESULTS: Steady-state plasma concentrations of tamoxifen and N-desmethyl tamoxifen (NDMT) were 5-10 microM following oral tamoxifen administration at 600 mg/m2 every 12 h for 7 days to normal and tumor-bearing dogs. Mild-moderate gastrointestinal toxicity (diarrhea, anorexia) and reversible neurotoxicity were observed in dogs receiving chemotherapy plus high-dose tamoxifen. Myelosuppression was not affected by combined treatment in tumor-bearing dogs. High-dose tamoxifen decreased the clearance and volume of distribution of full-dose doxorubicin. CONCLUSIONS: Concentrations of tamoxifen/ NDMT sufficient to inhibit Pgp may be achieved in dogs receiving full-dose chemotherapy with a moderate but acceptable increase in gastrointestinal toxicity. Tamoxifen affects doxorubicin metabolism in dogs at high doses resulting in increased serum exposure. Pharmacologic manipulation of Pgp expression or function in normal and tumor tissue in dogs may facilitate investigation of novel anticancer treatment strategies in humans.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1998

Determination of cefoxitin in serum and tissue.

A simple, rapid and sensitive method for the clean-up and analysis of cefoxitin in serum and tissue is described. Serum (0.5 ml) and tissue (100 mg) samples after homogenization underwent high speed centrifugation. Chromatography was performed on a muBondapak C18 cartridge using a mobile phase of 0.005 M potassium dihydrogen phosphate-acetonitrile-glacial acetic acid (77.5:22:0.5, v/v/v) with a flow-rate of 2.0 ml/min. Ultraviolet detection occurred at 235 nm. The procedure produced a linear curve for the concentration range 100-5000 ng/ml. The assay produced accurate, repeatable and rapid results for both tissue and serum samples without the need for chemical extraction.

Animals↗
PubMed1998

Black Creek Canal Virus infection in Sigmodon hispidus in southern Florida.

A total of 1,500 small mammals were collected and tested for antibodies cross-reactive to Sin Nombre virus (Hantavirus: Bunyaviridae) at 89 sites in a 1,600 km2 study area of southern Florida. More than 95% of the 123 seropositive animals were cotton rats (Sigmodon hispidus), suggesting infection by Black Creek Canal Virus, although seroreactive Rattus rattus (5 of 294) and Peromyscus gossypinus (1 of 39) also were captured. Crude seroprevalence in S. hispidus was 11%. Seroprevalence increased with body size and was more common in male (18%; n=451) than in female (6%; n=593) cotton rats. Infection within S. hispidus populations was widespread throughout the study area. Prevalence ranged from 0% to 60% at sites where more than five cotton rats were sampled but was not only a function of sample size. Sites with seropositive cotton rats were geographically clustered compared with sites with no seropositive cotton rats. Clustering was not due to the spatial distribution of sites with few animals, season of collection, or sex bias of animals captured at these sites. However, sites with no seropositive animals had an excess of animals in the intermediate size class (60-99 g) and a deficit of the largest and smallest animals. These data suggest that population structure within the habitat mosaic may play a significant role in the spatial distribution of hantavirus infection in local populations of reservoir species.

Animals↗
PubMed1997

Microassay for determination of itraconazole and hydroxyitraconazole in plasma and tissue biopsies.

A simple, rapid and sensitive method for the extraction and HPLC analysis of itraconazole and hydroxyitraconazole in tissue and plasma or serum is described. Tissue (5-100 mg) and plasma (0.1 ml) underwent a simple extraction into methanol. Chromatography was performed on a Novapak C18 column using a mobile phase of water-acetonitrile-diethylamine (42:58:0.05, v/v), pH 2.45, with a flow-rate of 1.5 ml/min. Fluorescence was measured at excitation 260 nm and emission 365 nm. The procedure produced a linear curve for the concentration range 10-1000 ng/ml. The development of the assay produced accurate, rapid repeatable results for both tissue and plasma or serum.

Animals↗
PubMed1996

Pharmacokinetics of pyropheophorbide-a-hexyl ether in the dog.

BACKGROUND AND OBJECTIVES: Pyropheophorbide-a-hexyl ether (HPPH) is a new compound being investigated for use as a photosensitizer for photodynamic therapy; however, the pharmacokinetics are not known for any of the target species likely to be treated with this drug. The objective of this study was to determine the pharmacokinetic parameters of this drug prior to institution of a clinical trial in canine patients with various cancers. STUDY DESIGN, MATERIALS AND METHODS: HPPH (0.3mg/kg i.v.) was administered to 12 dogs and blood samples were drawn at intervals for 24 hours and plasma HPPH concentrations were determined. Pharmacokinetic parameters were calculated for each dog. RESULTS: No evidence of toxicity was noted in any dog. The mean half-life was calculated to be 26.98 +/- 2.35 hrs. The mean clearance was 5.061 +/- 0.214 ml/hr/kg. The mean volume of distribution of the central compartment was 0.069 +/- 0.003 L/kg, and the mean steady state volume of distribution was 4.47 +/- 0.25 L/kg. CONCLUSION: The conclusion is that 0.3 mg/kg HPPH injected intravenously resulted in measurable plasma levels for 24 hrs, and resulted in no detectable adverse reactions.

Animals↗
PubMed1995

Immunomodulation by LFA3TIP, an LFA-3/IgG1 fusion protein: cell line dependent glycosylation effects on pharmacokinetics and pharmacodynamic markers.

LFA3TIP, a fusion protein comprised of the first extracellular domain of LFA-3 fused to the hinge, CH2, and CH3 domains of human IgG1 inhibits responses of human and non-human primate T cells in vitro. In seeking to optimize the expression efficiency to prepare large quantities of LFA3TIP for primate studies, the protein was produced in both the CHO (Chinese hamster ovary) and murine NS-0 myeloma cell lines. Although LFA3TIP derived from these cell lines performs identically in vitro in CD2 receptor binding and T cell assays, examination of a pharmacodynamic marker-the reduction in CD2+ lymphocyte numbers-following the administration of equal doses of NS-0 or CHO derived LFA3TIP to baboons, suggested that the effect of the NS-0 derived material was less sustained. Pharmacokinetic analysis of the materials in baboons and mice shows that LFA3TIP produced by NS-0 cells is rapidly cleared from circulation relative to the product derived from CHO cells. The disparate clearance profiles correlate with distinct glycosylation patterns, with LFA3TIP derived from NS-0 cells being less extensively sialylated than that from CHO cells due in part to alpha-galactosyl capping of selected lactosamine moieties in the N-linked glycans of NS-0 derived LFA3TIP. Moreover, enzymatic desialylation of CHO derived LFA3TIP results in a glycoprotein with an evanescent serum profile when administered to mice and baboons. These results correlate the extent of N-acetylneuraminic acid capping with the clearance rates of LFA3TIP derived from the two distinct cell lines, and underscore the importance of evaluating glycosylation dependent PK parameters when choosing production cell lines for recombinant immunotherapeutics.

Adjuvants, Immunologic↗
PubMed1994

High-performance liquid chromatographic determination of intrathecally administered [D-Ala2-D-Leu5]-enkephalin concentrations in canine cerebrospinal fluid.

A rapid and useful method for high-performance liquid chromatographic analysis of exogenous [D-Ala2-D-Leu5]-enkephalin (DADLE) in cerebrospinal fluid (CSF) is described. CSF (0.5 ml) samples were filtered using a 0.22-micron Co-Star filter. Chromatography was performed on a mu Bondapak C18 column using a mobile phase of A, 0.05 M sodium phosphate (monobasic, pH 6.0) and B, 60% acetonitrile in 0.05 M sodium phosphate (pH 6.0) with a flow-rate of 1 ml/min. Absorbance at 210 nm was measured. The procedure produced a linear curve for the concentration range 1-10 micrograms/ml. The development of the assay produced rapid, repeatable and accurate results for CSF analysis of DADLE at concentrations achieved with therapeutic administration of the peptide. This method could also be used in the future for analysis of compounds like DADLE.

Animals↗
PubMed1991

Cloning and expression of the cDNA for human gamma-glutamyl carboxylase.

The cDNA for human gamma-glutamyl carboxylase, which accomplishes the post-translational modification required for the activity of all of the vitamin K-dependent proteins, was cloned. The enzyme is a 758-residue integral membrane protein and appears to have three transmembrane domains near its amino terminus. The hydrophilic COOH-terminal half of the carboxylase has 19.3 percent identity with soybean seed lipoxygenase. Expression of the cloned cDNA resulted in an increase in carboxylase activity in microsomes of transfected cells compared to mock-transfected cells.

Amino Acid Sequence↗
PubMed1991

Determination of adriamycin in plasma and tissue biopsies.

A simple, rapid and sensitive method for the extraction and high-performance liquid chromatographic analysis of adriamycin in tissue and plasma is described. Tissue (5-100 mg) and plasma (1 ml) samples underwent a C18 Sep-Pak extraction into methanol. Chromatography was performed on a muBondapakphenyl column using a mobile phase of acetonitrile-0.1 M ammonium formate (pH 4.0) with a flow-rate of 2 ml/min. Fluorometric detection was used with an excitation of 480 nm and an emission of 550 nm. The procedure produced a linear curve for the concentration range 25-1000 ng/ml. The development of the assay produced rapid, repeatable and accurate results for both small tissue samples and plasma.

Animals↗
PubMed1991

Angiotensin converting enzyme system in the normal canine eye: pharmacological and physiological aspects.

The function of the renin-angiotensin-aldosterone system (RAAS) in the mammalian eye remains unclear, although alterations in the concentrations of various pathway components can influence intraocular pressure and the electroretinogram. Angiotensin Converting Enzyme (ACE) has been localized to ocular tissues and fluids. Aqueous humor and serum values of ACE are increased in sarcoid uveitis patients. We used the dog to simultaneously examine the effects of a topically administered ACE inhibitor on the intraocular pressure (IOP), on components of the renin-angiotensin pathway in the serum and aqueous humor, and to monitor any systemic effects of the ACE inhibitor. The novel ACE inhibitor, SCH 33861 (Schering Corporation), decreased IOP in amounts similar to timolol when applied topically to the canine eye. Serum ACE values significantly decreased in SCH 33861 treated dogs, while aqueous ACE values were only slightly decreased. A decrease in heart rate, and systolic and diastolic blood pressure in these dogs during the treatment period indicated probable systemic absorption. Normal values of aqueous humor and serum angiotensin-I were established for the dog. Plasma renin activity and angiotensin-I values were not significantly changed for any of the treatment groups. Topical application of SCH 33861 to the canine eye is a useful model to further evaluate the role of the renin-angiotensin system in the eye.

Administration, Topical↗
PubMed1990

Peroxidation-induced perturbations of erythrocyte lipid organization.

Peroxidation of erythrocyte membrane lipids by hydrogen peroxide perturbs the lipid bilayer and increases phagocytosis by macrophages. This study addresses the underlying mechanism of these processes, and in particular the role of malondialdehyde, a major byproduct of lipid peroxidation. When erythrocytes were treated with hydrogen peroxide or ascorbate/iron to generate malondialdehyde, or with malondialdehyde itself, only those cells treated with hydrogen peroxide showed increased phospholipid spacing and enhanced phagocytosis. This result indicates that the alterations observed are unique to hydrogen peroxide treatment, and that malondialdehyde does not play a role in inducing these changes in surface properties. Comparison of adherence to human umbilical vein endothelial cells and phagocytosis showed that increased phagocytosis was not mirrored by enhanced adherence. This result suggests that two different signals may mediate recognition of erythrocytes by macrophages and by endothelial cells.

Cell Adhesion↗
PubMed1989

Mapping of monoclonal antibodies to human factor IX.

We used recombinant DNA techniques to map a panel of six monoclonal antibodies (MoAbs) to regions of the human factor IX molecule. A-2 maps to 17 amino acids at the amino terminus of the heavy chain of IXa; 2D5, an inhibitor of clotting, is defined to 36 amino acids of the first EGF-like domain of human factor IX. A-4, A-5, C10D, and FXC008 all map to a region of the heavy chain containing amino acids 180 through 310, suggesting an immunodominant site. FXC008 has been reported to interfere with binding of factor IXa to factor VIII:Ca.

Amino Acid Sequence↗
PubMed1988

Genetic defect responsible for the dysfunctional protein: factor IXLong Beach.

DNA sequence analysis of the gene coding for the variant protein, factor IXLong Beach (FIXLB), has identified a transition mutation in an otherwise normal factor IX (FIX) gene. Genomic DNA clones spanning 35 kilobase (kb) pairs of the FIXLB gene were isolated. A gene analysis strategy that specifically characterized exons and their flanking intron sequences predicted the entire amino acid sequence of FIXLB. A thymine to cytosine transition causes the substitution of a threonine codon (ACA) for an isoleucine codon (ATA) in exon VIII of the FIXLB gene. This mutation results in an amino acid substitution at residue 397 of the FIX zymogen and the phenotypic display of hemophilia-B. Previous studies revealed that activated purified FIXLB (FIXaLB) had normal Ca2+, phospholipid, and factor VIIIa binding characteristics. However, FIXaLB activated factor X or factor VII (with their cofactors Ca2+ and phospholipid) at significantly reduced rates, suggesting that the defect in FIXaLB lies near or within the catalytic triad of the FIX heavy chain. Identification of an amino acid substitution near the carboxy-terminus of the FIXaLB heavy chain supports the earlier characterization of this variant protein. Moreover, our data identify a residue in the catalytic domain of FIXa essential for normal function.

Base Sequence↗