Relationship between feline leukemia virus antigen expression and viral infectivity in blood, bone marrow, and saliva of cats.

Correlation was greater than 90% between feline leukemia virus (FeLV), group-specific antigen (GSA) in leukocytes, and viral infectivity (VI) in serum or plasma from 132 cats infected with either the Rickard strain of FeLV, the Snyder-Theilen strain of feline sarcoma virus, or field strains of FeLV. Detection of GSA in blood cells was at least as sensitive as detection of VI in serum. In 45% of FeLV GSA-positive cats inoculated with FeLV-Rickard strain, VI was detected in saliva. No saliva samples from GSA-negative cats had VI. Sequential bone marrow biopsies from 34 cats inoculated with Snyder-Theilen feline sarcoma virus indicated that the correlation between FeLV GSA in bone marrow cells and blood cells was virtually 100%. FeLV GSA appeared in bone marrow leukocyte precursors 1 week before its appearance in peripheral blood leukocytes in 50% of the cats. The FeLV GSA-positive state was transient (3 to 6 weeks) in 34% of the Snyder-Theilen feline sarcoma virus-inoculated cats.

Tyrosine kinase phosphorylation of GABAA receptors.

Phosphorylation of purified bovine brain GABAA receptors by the tyrosine kinase, pp60v-src was examined. pp60v-src phosphorylated two bands of 54-62 kDa and 48-51 kDa that migrated to approximately the same position as bands recognized by antisera against the beta 2 and gamma 2 GABAA receptor subunits, respectively. Bacterially expressed proteins containing the putative large cytoplasmic loops of the beta 1 and gamma 2L subunits were phosphorylated by pp60v-src, indicating that the phosphorylation sites are located in these subunit domains. The tyrosine kinase inhibitors, genistein and the tyrphostins B-42 and B-44, inhibited muscimol-stimulated 36Cl- uptake in mouse brain membrane vesicles (microsacs). magnitude of the tyrphostin B-44-induced inhibition of muscimol-stimulated 36Cl- uptake was significantly reduced in microsacs that were lysed and resealed under conditions that inhibit phosphorylation. GABA-gated Cl- currents were also inhibited by genistein and tyrphostin B-44 in Xenopus oocytes expressing alpha 1 beta 1 and alpha 1 beta 1 gamma 2L subunits. Consequently, protein tyrosine kinase-dependent phosphorylation appears to be another mechanism of regulating the function of GABAA receptors.

Plasmid content of some oral microorganisms isolated from subgingival plaque.

Eighty-five strains of bacterial species selected from the predominant cultivable dental plaque flora of patients with different periodontal pathologies were examined for their plasmid content. Microorganisms studied included: Actinomyces viscosus, A. odontolyticus, Bacteroides asaccharolyticus (B. gingivalis), B. melaninogenicus subspecies intermedius, and subspecies melaninogenicus, Capnocytophaga ochracea (B. ochraceus), and Fusobacterium nucleatum. Three B. melaninogenicus isolates showed plasmids of approximately 2.7-2.9 Mdalton (mega-dalton) molecular size. Restriction enzyme digests of the plasmids demonstrated dissimilar patterns when electrophoresed on agarose gels. In other microorganisms, including the Actinomyces species, plasmids were not observed.

Parental imprinting and Angelman syndrome.

Angelman syndrome is an inherited disorder that includes severe mental retardation and epilepsy. Patients have no speech, puppet-like gait with jerky movements, hyperactivity, disturbed sleep, bouts of inappropriate laughter, a pronounced jaw, and widely spaced teeth. The syndrome results from deletion or mutation within maternal chromosome 15q11-q13. Considerable evidence suggests that the gene or genes responsible for Angelman syndrome are expressed only from the maternal chromosome 15, a situation known as parental imprinting. This epigenetic marking of certain regions of the parental genomes is characterized by parent-of-origin-specific allelic DNA methylation, allele-specific DNA replication timing, and physical pairing of the two chromosome 15 homologues. Imprinting is important for normal development, and its disregulation causes several human disorders. The epilepsy of Angelman syndrome has been studied and indicates a rather typical electroencephalographic abnormality with slowing and notched wave and spikes. Various types of seizures occur, usually including myoclonus and atypical absence. Variable severity among patients suggests potential molecular diversity in the genetic mechanism, possibly the involvement of more than one gene. Angelman syndrome can arise from the following molecular genetic defects: a deletion in 15q11-q13 that covers the Angelman gene or genes, mutations that alter imprinting, and paternal uni-parental disomy for the region. Another 20% or so of patients with clinical symptoms of Angelman syndrome have none of these three defects but are believed to have mutations in one or more genes in the region, and this may be familial. The UBE3A gene, which codes for the enzyme ubiquitin protein ligase involved in protein degradation and processing, has been found to be mutated in many but not all of patients with Angelman syndrome and can be considered a major Angelman candidate gene. Other potential candidate genes in the region include a cluster of three GABAA receptor subunits, which are involved in inhibitory synaptic transmission in the brain. The GABRB3 gene, which codes for the beta 3 subunit, is deleted in most persons with Angelman syndrome. The absence of this gene in mice causes craniofacial abnormalities and neurologic impairment with seizures. The exact role of UBE3A and GABRB3 in the syndrome and their imprinting status are under investigation.

Density-gradient sedimentation of feline bone marrow cells with polyvinyl-pyrrolidone-coated silica gel.

Feline bone marrow cells were separated based on their density distribution in a continuous polyvinyl-pyrrolidone-coated silica gel density gradient. Morphologic identification of cytocentrifuged preparations at specific densities revealed a progressive increase in density with maturation of the cells within the granulocytic and erythroid series. Segmented eosinophils peaked at 1.084 g/ml, monocytes at 1.07 g/ml, and lymphocytes spanned the density of 1.068 to 1.084 g/ml. Bone marrow samples from 6 healthy specific-pathogen-free cats were separated by the continuous polyvinyl-pyrrolidone-coated silica gel gradient and were studied for progenitor colony formation in methylcellulose. Erythroid colony formation was greatest at a density of 1.084 g/ml and also appeared in cells from 4 cats at a lighter density of 1.016 to 1.05 g/ml. Colony formation in the granulocytic series revealed progenitors throughout the gradient with enrichment at 1.055 g/ml.

Bone ingrowth into porous silicon nitride.

Achieving solid skeletal attachment is a requirement for the clinical success of orthopedic implants. Porous or roughened surfaces and coatings have been developed and used with mixed success to achieve attachment due to bone ingrowth. Silicon nitride is a high performance ceramic whose strength, imaging properties, and biocompatibility make it a candidate material for orthopedic implants. A porous form of silicon nitride, cancellous-structured ceramic (CSC), has been developed. CSC is a nonresorbable, partially radiolucent porous structure that can be bonded to orthopedic implants made of silicon nitride to facilitate skeletal attachment. The purpose of this study was to quantify the extent and rate of bone ingrowth into CSC in a large animal model. Cylindrical implants were placed bilaterally using staged surgeries in the medial femoral condyle of six sheep. Condyles were retrieved after 3 and 6 months in situ and prepared for examination of bone growth under SEM. Bone grew into CSC to extents and at rates similar to those reported for other titanium porous surfaces in studies involving large animals and postmortem retrievals in humans. Bone ingrowth was observed at depths of penetration greater than 3 mm in some implants after only 12 weeks in situ. Bone ingrowth into CSC is a viable method for achieving skeletal attachment.

A versatile and easily fabricated mouthstick.

More and more handicapped individuals may be seeking dental treatment. Treatment may include construction of special devices to help these patients lead more productive and fulfilling lives. A mouthstick has been designed that can be constructed from readily available parts. The versatility and decreased cost of the mouthstick make it readily available to these needy patients.

Adoptive immunotherapy of feline leukemia virus infection using autologous lymph node lymphocytes.

Adoptive immunotherapy using autologous cells expanded ex vivo from lymph nodes was examined in cats infected with the retrovirus feline leukemia virus (FeLV). Cells were obtained from popliteal lymph nodes from 18 FeLV-antigen-positive cats without complications; a mean of 6.2 x 10(7) cells were obtained. Lymph node cells were cultured with 600 IU/ml interleukin-2 (IL-2) for 7 days. Cells expanded 0.8- to 11-fold (mean, 2.7; median, 2.4); were 80% +/- 8.0% CD3+, 29% +/- 8.1% CD4+, and 41% +/- 7.0% CD8+, and exhibited cytolytic activity against FeLV-transformed FL74 cells. Sixteen cats received a single intravenous infusion of 0.13 to 3.9 x 10(8) cells. Cell infusion was well tolerated; fever developed approximately 1 hour postinfusion. Clinical activity, antiviral activity, or both was observed in 10 cats. Nine cats had clinical responses with improvement in weight, activity, appearance, or a combination of these that began 2 to 4 weeks after cell infusion and that lasted for up to 13 or more months. FeLV antigen became undetectable in 4 cats. These results indicate that adoptive immunotherapy using autologous lymph node cells, activated and expanded ex vivo in short-term cultures with low concentrations of IL-2, can modulate the course of a retroviral infection.

Enamel reduction and the bond strength of resin-bonded retainers.

The authors investigated the shear bond strength of etched enamel retainers bonded to enamel reduced by different amounts. The depth of reduction in this study had no influence on bond strength.

Convulsant/barbiturate activity on the soluble gamma-aminobutyric acid-benzodiazepine receptor complex.

Cage convulsant t-butyl bicyclophosphoro[35S]thionate binding activity in rat brain membrane homogenates was solubilized with the zwitterionic detergent 3-[(3-cholamidopropyl)-dimethylammonio]propane sulfonate (Chaps) and shown to co-purify with the benzodiazepine--gamma-aminobutyric acid (GABA) receptor complex on gel filtration and affinity chromatography. Whereas convulsant binding activity, but not GABA and benzodiazepine receptor binding, was eliminated by solubilization in other detergents like sodium deoxycholate or Triton X-100, or by addition of Triton X-100 to the extracts solubilized in the zwitterionic detergent, convulsant activity was not irreversibly lost or selectively unstable, but could be restored by exchanging the protein back into the detergent Chaps. The GABA-benzodiazepine receptor activity solubilized in Chaps alone, containing convulsant activity, and a sample in Chaps supplemented with Triton X-100 and lacking convulsant activity, did not differ in size as measured by gel filtration column chromatography or by radiation inactivation target size analysis. This suggests that convulsant binding activity does not require any additional protein subunits or other macromolecules nor any unique aggregation state relative to GABA and benzodiazepine receptor binding, and that all three activities reside on the same protein complex. As in intact brain, the target size for convulsant binding activity was 3-5 times that of benzodiazepine binding activity, suggesting that an oligomeric protein structure of the receptor complex with intact strong subunit interactions present in the native membrane environment is needed for convulsant activity, and that this and other properties are more preserved in Chaps than in other detergents.

Serologic response to canine distemper viral antigens in gnotobiotic dogs infected with canine distemper virus.

The humoral immune response to canine distemper viral antigens in gnotobiotic dogs experimentally infected with canine distemper virus was studied by the complement fixation and serum neutralization tests. Antibody titers measured by both serologic methods varied inversely with the severity of disease produced. Recovered dogs demonstrated the highest titers of antibody, whereas fatally infected dogs had little or no antibody activity in their sera. A third group of dogs, characterized by chronic persistent infection, had intermediate levels of anitbody to canine distemper virus. Preliminary characterization of the viral antigens involved in the complement fixation test indicated that at least two antigenic components were involved. One antigen was soluble in ether and was heat-labile, whereas the other was relatively heat-stable and was unaffected by treatment with ether. No evidence for serologic cross-reaction between viral and central nervous system components was found. A vigorous antibody response to envelope antigen determinants in addition to core determinants distinguished immune dogs from persistently infected dogs. The results of this study suggest that the inability to produce antibodies to envelope antigens may be a crucial factor in the establishment of a persistent infection with canine distemper virus in these dogs.

Suppression of in vitro neutrophil function by feline leukaemia virus (FeLV) and purified FeLV-p15E.

Feline neutrophils (PMN) were isolated and exposed to ultraviolet light-inactivated feline leukaemia virus (UV-FeLV) and purified envelope component p15E (FeLV-p15E). Functional capacity of exposed PMN was measured in vitro utilizing the chemiluminescence (CL) response. PMN exposed to UV-FeLV demonstrated depressed CL responses to Ca2+-ionophore A23187 and latex particles. However, FeLV-p15E produced significant suppression in the CL response to A23187 but failed to produce significant alterations in response to latex particles. The data indicate that FeLV-p15E may, in part, be responsible for increased morbidity and mortality among FeLV-infected cats through suppression of the PMN population.

Examination gloves as barriers to hand contamination in clinical practice.

OBJECTIVE: To test the effectiveness of vinyl and latex gloves as barriers to hand contamination with gram-negative organisms and enterococci during routine hospital procedures. DESIGN AND INTERVENTIONS: We studied 137 procedures during which a health care worker's gloved hand contacted a patient's mucous membrane and was thus potentially contaminated with gram-negative rods or enterococci. Quantitative hand cultures were obtained from each health care worker before and after the gloved contact using a modified glove juice method, and the exterior glove surface was also quantitatively cultured after patient contact. Used gloves were then tested for leaks using the American Society for Testing and Materials' watertight test. SETTING: Harborview Medical Center, a 330-bed city-county hospital and level I regional trauma and burn center, is both a teaching facility affiliated with the University of Washington and the major provider of care to indigent and uninsured persons in Seattle-King County, Washington. PATIENTS AND OTHER PARTICIPANTS: Respiratory therapists performing endotracheal tube care on intubated intensive care unit patients, registered nurses performing digital rectal stimulation for bowel training on patients with spinal cord injury in the rehabilitation ward, and dentists performing routine dental examinations and procedures on healthy outpatients in the dental clinic. MAIN OUTCOME MEASURE AND RESULTS: Eighty-six of the 135 gloves cultured had gram-negative rods or enterococci on the external surface after use and were thus sources of potential hand contamination. Microbial contamination of the health care worker's hands occurred in 11 (13%; 95% confidence interval, 6% to 20%) of these 86 events, and was more frequent with vinyl (10 of 42) than latex (one of 44) gloves (P < .01). After use, glove leaks were also more frequent in vinyl gloves (26 of 61) than with latex gloves (six of 70) (P < .001). Even when leaks were present, gloves prevented hand contamination in 77% of instances and quantitative counts of microorganisms contaminating hands were 2 to 4 logs less than counts on external glove surfaces. Health care workers reported awareness of the presence of glove leaks in only seven (22%) of the 32 events in which leaks were subsequently demonstrated. CONCLUSIONS: Under conditions of routine use, gloves effectively function as a protective barrier even when leaks are present. Latex gloves were less frequently associated with leaks and hand contamination. Since hand contamination occurred after 13% of exposures and cannot be readily identified by health care workers, routine hand washing should be done after each patient contact.

Saliva as a source of feline leukemia virus antigen for diagnosis of disease.

An enzyme-linked immunosorbent assay (ELISA) for detection of feline leukemia virus (FeLV) p27 in saliva was tested for its accuracy and sensitivity in diagnosing FeLV infections. Saliva and serum samples from 564 clinical cases were tested with a 99.2% specificity. The overall accuracy of the saliva ELISA reactive to the serum ELISA was 97.9%. Experimentally, the ELISA saliva was the least sensitive in diagnosing early FeLV infections. However, the overall accuracy, ease of use, and simplicity of the test support its use as a screening procedure in clinical practice.