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Biomedical subjects

C A Davis

Publications and source records attributed to C A Davis.

At least 73 records · Page 4Linked to original sources

Expression patterns of the homeo box-containing genes En-1 and En-2 and the proto-oncogene int-1 diverge during mouse development.

We have compared the expression of the murine genes En-1,En-2, and in-1 during development by in situ hybridization. Expression of all three genes was first detected at 8.0 days in overlapping bands of the anterior neural folds. By 12.0 days the expression patterns diverged. En-1 and En-2 were expressed in a similar ring of cells in the central nervous system (CNS) at the midbrain/hindbrain junction. En-1 was also expressed de novo in two lateral stripes extending the length of the hindbrain and spinal cord, in the developing vertebral column, in two lateral stripes of dermatome-derived cells, and in the tail and limb buds. By 12.0 days int-1 expression showed little overlap with the En genes and could not be detected at later stages. At 15.5 days En gene expression was primarily limited to the midbrain/hindbrain in overlapping but nonidentical sets of differentiated cells. In the adult, En-1 and En-2 marked the same sets of cells in the pons, but En-2 alone was detected in the granular layer of the cerebellum. The results are consistent with int-1 and the En genes playing a role early in development in defining spatial domains in the CNS. Later in development the En genes may have an additional function in neurogenesis. En-1 expression in the developing pericordal tube suggests that it may also be involved in vertebral assembly.

Animals↗

Expression of the homeo box-containing gene En-2 delineates a specific region of the developing mouse brain.

We have examined the pattern of expression of the homeo box-containing gene En-2 during mouse embryogenesis using in situ hybridization. Transcripts were first detected in the neural folds of 8.0-day, 5-somite embryos, and expression continued throughout development into adulthood. Hybridization occurred only in the central nervous system (CNS) and was limited to one band of the neural tube and to parts of those structures that later developed from it; the cerebellum, pons, periaqueductal gray, and colliculi. Expression in the germinal zone of the CNS was uniform within the hybridizing band. However, later in development, once cells had migrated out of the germinal zone, there was a reduction in the extent of hybridization and an increase in its spatial complexity. In the adult, expression of En-2 appeared to be limited to specific groups of neurons. The early, localized expression of En-2 within an apparently homogeneous tissue is consistent with the hypothesis that En-2 plays a role in defining a spatial domain within the developing brain.

Animals↗

Cloning and expression of a Streptococcus sanguis surface antigen that interacts with a human salivary agglutinin.

Human saliva contains a high-molecular-weight glycoprotein (agglutinin) which binds to specific streptococci in a calcium-dependent reaction leading to the formation of bacterial aggregates. We report the cloning of a gene encoding a surface antigen from Streptococcus sanguis M5 and show that the expressed protein inhibits agglutinin-mediated aggregation and specifically binds the salivary agglutinin in a calcium-dependent fashion. Clones isolated from the immunological screening of S. sanguis M5 genomic libraries with polyclonal antibodies against whole cells were assayed for the ability to compete with S. sanguis for agglutinin. One clone, pSSP-5, expressed antigens of 165 and 130 kilodaltons (kDa) possessing this activity. A 3-kilobase-pair (kbp) insert fragment from this clone was used to screen a genomic library in lambda EMBL3 which resulted in the isolation of clone SSP-5A. This clone contained an insert of 17 kb and expressed proteins of 170 to 205 kDa that reacted with the anti-S. sanguis antibodies. Subcloning of a 5.3-kbp EcoRI-BamHI fragment from SSP-5A produced pEB-5, which expressed streptococcal components that were indistinguishable from SSP-5A. The streptococcal antigen was purified by gel permeation and ion exchange chromatography and shown to potently compete with S. sanguis M5 cells for agglutinin. The antigen also bound purified salivary agglutinin in the presence of 1 mM CaCl2. This binding was inhibited by EDTA. Both the SSP-5 antigen and a 205-kDa protein in surface protein extracts from S. sanguis M5 cross-reacted with antibodies directed against antigen B from S. mutans and SpaA from S. sobrinus 6715. These results indicate that a 205-kDa surface protein that is antigenically related to SpaA and antigen B is involved in the binding of salivary agglutinin to S. sanguis M5.

Agglutinins↗

Altered disposition of propylthiouracil in cats with hyperthyroidism.

The oral and intravenous disposition of the anti-thyroid drug propylthiouracil (PTU) was determined in six clinically healthy cats and four cats with naturally occurring hyperthyroidism. Compared with the normal cats, the mean plasma elimination half-life of PTU was significantly (P less than 0.001) shorter in the hyperthyroid cats (77.5 +/- 5.8 minutes compared with 125.5 +/- 3.7 minutes) and the total body clearance of PTU was significantly (P less than 0.05) more rapid in the cats with hyperthyroidism (5.1 +/- 0.8 ml kg-1 min-1 compared with 2.7 +/- 0.2 ml kg-1 min-1). Following oral administration, both the bioavailability (59.7 +/- 4.9 per cent compared with 73.3 +/- 3.7 per cent) and peak plasma concentrations (14.5 +/- 1.6 micrograms ml-1 compared with 18.9 +/- 0.9 micrograms ml-1) of PTU were significantly (P less than 0.05) lower in the hyperthyroid cats than in the control cats. No difference was noted, however, between the apparent volume of distribution for PTU in the two groups of cats. Overall, results of this study indicate that the oral bioavailability of PTU is decreased and PTU disposition is accelerated in cats with hyperthyroidism.

Animals↗

Lipid/water ratio of bone marrow measured by phase-encoded proton nuclear magnetic resonance spectroscopy.

Proton NMR spectroscopy (spatially resolved) at 0.7 Tesla with a whole body superconducting magnet was performed on bone marrow from human lumbar spine to measure the lipid/water ratio. Phase-encoded proton spectroscopy was performed by using (1) a spin-echo sequence with selective 90 degrees and 180 degrees rf pulses on the Z and X axes; (2) 64 different phase encodes along the Y axis; and (3) echo acquisition with no gradient. The lipid/water ratios also were measured by the Folch chemical extraction technique. The set of phase-encoded spectra obtained from the spine demonstrates the chemical difference between the vertebral disk (large water peak and no lipid peak) and the marrow (smaller water peak and detectable lipids). Lipid/water ratios were lower than the chemically measured ratios. The phase-encoded proton spectroscopic technique presented in this study allowed rapid acquisition of localized spectra from the bone marrow in each of three vertebral bodies of the intact human spine. This technique provided a measure of the mobile lipid/water ratio weighted by relaxation times.

Body Water↗

A randomized phase I/II study of continuous versus intermittent intravenous interferon gamma in patients with metastatic melanoma.

Thirty patients with documented metastatic melanoma were randomly assigned to receive recombinant DNA-produced gamma-interferon (specific activity approximately, 20 MU/mg) intravenously (IV) over either two or 24 hours at dosages of 3, 30, 300, 1,000, or 3,000 micrograms/m2. Objective toxicity resembled that of alpha-interferon and included fever, chills, myalgias, headache, and fatigue. Neutropenia, elevations in liver enzymes, tachyarrhythmias, and CNS changes also were noted. Dose-limiting toxicity included neutropenia, liver enzyme abnormality, constitutional symptoms, and a change in mental status. The incidence of toxicity was qualitatively similar in both two- and 24-hour treatment arms, but was quantitatively more severe in the 24-hour continuous infusion arm. Maximum tolerated dose was 1,000 micrograms/m2 in both schedules. Pharmacokinetic studies showed a half-life of six to nine hours. One patient had a complete response after two cycles of therapy and an additional patient entered partial remission after three cycles. Recombinant gamma-interferon (rIRN-gamma) is tolerated at dosages of 1,000 micrograms/m2 administered daily either by two or 24 hour infusion for 14 days in patients with metastatic melanoma. The responses documented in this early trial warrant further evaluation for the treatment of metastatic melanoma.

Adult↗

The effects of bone on proton NMR relaxation times of surrounding liquids.

Preliminary attempts by our group at UCSF to assess fat content of vertebral marrow in the lumbar spine using relaxation time information demonstrated that the presence of trabecular bone affects relaxation times. The objective of this work was a thorough study of the effects of bone on NMR relaxation characteristics of surrounding liquids. Trabecular bone from autopsy specimens was ground up and sifted into a series of powders with graded densities ranging from 0.3 gm/cc to 0.8 gm/cc. Each powder was placed first in n-saline and then in cottonseed oil. With spectroscopy, spin-lattice relaxation times (T1) and effective spin-spin relaxation times (T2*) were measured for each liquid in each bone powder. As bone density and surface to volume ratio increased, T1 decreased faster for saline than for oil. T2* decreased significantly for both water and oil as the surface to volume ratio increased. It was concluded that effects of water on T1 could be explained by a surface interaction at the bone/liquid interface, which restricted rotational and translational motion of nearby molecules. The T1s of oil were not affected since oil molecules are nonpolar, do not participate in significant intermolecular hydrogen bonding, and therefore would not be expected to interact strongly with the bone surface. Effects on T2* could be explained by local magnetic field inhomogeneities created by discontinuous magnetic susceptibility near the bone surface. These preliminary results suggest that water in contact with trabecular bone in vivo will exhibit shortened relaxation times.

Bone Marrow↗

Procainamide in the dog: antiarrhythmic plasma concentrations after intravenous administration.

Procainamide hydrochloride was administered to ouabain-intoxicated dogs to determine an antiarrhythmic plasma concentration of procainamide. Ventricular arrhythmias were produced in dogs following intravenous injections of ouabain. After a sustained ventricular tachycardia was achieved, procainamide was administered and plasma samples collected for assay. Plasma procainamide was assayed by fluorescence polarization immunoassay. Procainamide was administered at increasingly higher constant rate infusions in order to achieve intermittent, steady-state plasma concentrations. Infusion rates were calculated on the basis of previous pharmacokinetic information. All six dogs that received procainamide converted to a normal sinus cardiac rhythm after attaining a mean plasma concentration of 33.8 micrograms/ml with a range of 48.5 micrograms/ml-25.0 micrograms/ml. It was observed that the computer-generated prediction of plasma concentrations based upon previous pharmacokinetic data produced an underestimate of the actual plasma concentrations. These data may suggest that plasma concentrations of procainamide for controlling some cardiac arrhythmias in dogs may be higher than plasma concentrations cited for human patients.

Animals↗

Pharmacokinetics of procainamide hydrochloride in dogs.

Pharmacokinetics of procainamide hydrochloride were studied in 2 groups of dogs. In a group of 6 dogs, procainamide was administered IV at a small dose of 8 mg/kg (group 1), and blood samples were obtained for 3.5 hours. In another group of 6 dogs, procainamide was administered IV and orally at an average dose of 25.5 mg/kg (group 2) in a crossover manner. Blood samples were obtained for 48 hours. In 2 dogs (previously used in part II), N-acetylprocainamide (NAPA) was administered IV at a dose of 10 mg/kg. Plasma samples were assayed for procainamide by fluorescence polarization immunoassay, and NAPA samples were assayed by high-performance liquid chromatography. In group 1, the elimination of procainamide was described by a 1-compartment, open pharmacokinetic model. The elimination half-life was 2.43 hours, the apparent volume of distribution was 1.44 L/kg, and the systemic clearance was 0.412 L/kg/hr. In group 2, 2 of the 6 dogs were described by a 1-compartment model, and 4 of the 6 dogs were described with a 2-compartment pharmacokinetic model. The elimination half-life for the IV dosage was 2.85 hours, the apparent volume of distribution was 2.13 L/kg, and the systemic clearance was 0.519 L/kg/hr. For the orally administered dose, the bioavailability was 85%, and the absorption half-life was 0.5 hours. There was no evidence of acetylation of procainamide to NAPA or deacetylation of NAPA to procainamide. The estimated elimination half-life of NAPA was 4.7 hours.

Acecainide↗

A gene family in Drosophila melanogaster coding for trypsin-like enzymes.

We have isolated a clustered gene family in D. melanogaster that codes for trypsin-like enzymes. The gene family has been localized to 47D-F by in situ hybridization to polytene chromosomes. The four genes in the family are transcribed in alternating orientations, and code for 1000 nt mRNAs. Transcripts are present at all stages of the life cycle. In situ hybridization to mRNA in tissue sections of third instar larvae showed that transcripts were restricted to the mid-gut. One gene was sequenced. The translated amino acid sequence of the proposed active enzyme is 42% homologous to bovine trypsin. Regions of functional importance are more strongly conserved. These include the active site residues asp102, his57, ser195, and the residue asp189 which is reputed to bind the basic residue at the substrate cleavage site. The activation peptide is not homologous to that of most vertebrate trypsins, suggesting a modified activation mechanism. The sequence further strengthens the hypothesis that the chymotrypsin cleavage specificity developed separately in the vertebrates and invertebrates.

Amino Acid Sequence↗

MR effect on murine spermatogenesis.

The cytotoxic effects of magnetic resonance (MR) on spermatogenesis in mice were studied. Mice were exposed to a strong static magnetic field from a 0.3-T MR imaging device. The effect of this treatment on different classes of spermatogenic cells was investigated by counting sperm heads at various times after exposure. Sixty-six hours of continuous exposure did not produce any significant cytotoxicity to these cells. This period of continuous exposure is far longer than any anticipated occupational exposure to humans. Thus, acute and subacute exposure to static magnetic fields associated with diagnostic use of MR imaging devices is unlikely to have any significant adverse effect on spermatogenesis.

Animals↗

Comparison of intramuscular and intravenous recombinant alpha-2 interferon in melanoma and other cancers.

In two phase I-II trials, 33 patients were given recombinant interferon alpha-2 daily at dosages of 3, 10, 30, 50, or 100 MU/d for up to 4 weeks by intramuscular or intravenous routes. Dose-limiting toxicities, including neutropenia, elevated hepatocellular enzyme levels, fatigue, and disturbed mentation, correlated with differing serum pharmacokinetics of interferon in the two trials. In the intramuscular study, dose-limiting toxicity occurred at all dosages greater than 10 MU/d, at a median of 6 to 9 days of treatment. In the intravenous dose-study, limiting toxicity was seen only at dosages of 100 MU/d, at a median of day 8. Twenty-three patients had metastatic melanoma and 4 had objective partial or complete responses at dosages of 10 to 50 MU/d in the first month. Two patients with complete responses are free of tumor after 2.5 years of follow-up. A fifth patient had delayed complete regression, requiring 1 year to achieve maximum response, but remains free of disease at 26 months since entry to the trial. Interferon had antitumor activity against melanoma by both routes tested, at dosages of 10 to 50 MU/d.

Adult↗

Circulating immune complexes and the nephropathy of cystic fibrosis.

To explore the putative nephropathic role of Pseudomonas-associated immune complexes, the authors measured the quantity of immune complexes in sera obtained, before death, from 20 patients with cystic fibrosis, and compared these findings with the histologic features of the lesions and with immunofluorescence patterns of kidney tissue obtained at autopsy. The immune complexes were measured by solid-phase C1q (C1q immune complex) and conglutinin to detect complexes containing IgM, IgA, and IgG. Elevated levels of C1q immune complex (13 patients) suggested the possibility of renal deposition of C3 (P less than 0.005) and IgM (P less than 0.05). The only three patients with IgA tissue deposits had elevated levels of C1q immune complex with normal IgA immune complexes. No other assay findings correlated with the immunofluorescence findings. Despite the prominent C3 in tissue deposits, the histologic features were not significantly associated with the results of the immune complex assays. This study indicates that complement-activating IgM-containing complexes can be deposited in renal tissues of patients with cystic fibrosis, but their nephropathogenicity is doubtful. These observations of kidney lesions, which diminish the injurious role of immune complexes in cystic fibrosis, may be relevant to an understanding of the pathogenesis of the lung lesions, which recent studies have linked to the presence of immune complexes.

Adolescent↗

Zinc supplementation during pregnancy: zinc concentration of serum and hair from low-income women of Mexican descent.

The effect of zinc supplementation on concentrations of zinc in hair and serum of 213 pregnant Hispanic women attending a clinic in Los Angeles was assessed using a random, double-blind experiment. Both the treatment (T) and control (C) groups received similar vitamin and mineral supplements except that 20 mg zinc was added to the supplements for the treatment group. Nutrient intakes were calculated from 24-h recalls. The initial mean dietary zinc intake of both groups was about 50% of the Recommended Dietary Allowance (9 +/- 5 mg). Initially there were no significant differences between the two groups in mean zinc levels in serum (66 +/- 11 micrograms/dl, C, and 65 +/- 12 micrograms/dl, T) or in hair (184 +/- 41 micrograms/g, C, and 175 +/- 38 micrograms/g, T). Zinc supplementation did not alter mean zinc levels in serum or hair but significantly (p less than 0.05) reduced the number of low serum zinc values (less than or equal to 53.3 micrograms/dl) toward the end of pregnancy. Although serum zinc levels do decline in pregnancy, our results suggest that severely depressed levels (less than or equal to 50 to 55 micrograms/dl) indicate inadequate zinc status.

Adolescent↗

Membranoproliferative glomerulonephritis and alpha 1-antitrypsin deficiency in children.

Two white girls had reduced serum concentration of alpha 1-antitrypsin (alpha-AT), phenotype ZZ, and liver disease. Hepatocytes exhibited the microscopic criteria of alpha-AT deficiency. Hypocomplementemia, elevated circulating immune complexes (patient 1), clinical signs of renal disease, and the histologic findings of membranoproliferative glomerulonephritis (MPGN) type I developed. Immunoglobulin A (but not alpha-AT) was demonstrable immunologically as a component of glomerular deposits in patient 1. Among 53 patients with MPGN but without clinical signs of liver disease, none had Pi type Z. Among 23 patients with phenotype ZZ but without clinical signs of kidney disease, six had abnormal complement protein levels, but the pattern did not resemble that of idiopathic MPGN type I. These results are consistent with the conclusion that MPGN in the two patients reported here is a consequence of their chronic liver disease and is not directly related to the presence of the allelic alpha-AT variant PiZ.

Adult↗

Correlations between ancillary assays and bacteriuria in children with myelodysplasia and ileal conduit urinary diversions.

Urine from children with ileal conduit urinary diversions was assayed for antibody-coated bacteria, lysozyme and lactic dehydrogenase isoenzymes, and correlated with the results of quantitative urine culture. A significant proportion of the antibody-coated bacteria and lysozyme assays were positive when more than 10(4) organisms per ml. of urine were found. Lactic dehydrogenase isoenzyme results were frequently at variance with the other assay and culture results. Because none of the ancillary assays was invariably positive in patients with symptoms suggestive of upper tract disease, they remain imperfect research tools for localization of urinary tract infections in patients with ileal conduits. In this patient population quantitative urine culture remains the best clinical method for diagnosing urinary tract infections and evaluating the adequacy of antibiotic therapy. However, since the antibody-coated bacteria and lysozyme assays converted to negative with successful treatment, these tests may provide additional guidance to the physician in assessing the adequacy of treatment of urinary tract infections in patients with ileal conduit diversions.

Adolescent↗