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Biomedical subjects

B R Miller

Publications and source records attributed to B R Miller.

At least 55 records · Page 3Linked to original sources
PubMed1994

Compliance with oral hygiene recommendations following periodontal treatment in client-owned dogs.

A telephone questionnaire survey of owners of 51 dogs was conducted. All 51 dogs had had periodontal treatment at VHUP six months or more previously. At that time, they had received a tooth brush, dentifrice, instructions and a demonstrations of brushing. The owners were asked to brush the dogs' teeth once daily. At the time of survey, 53% of the clients were still brushing several times a week and 38% were no longer brushing at all.

Animal Welfare↗
PubMed1993

Intramitochondrial sorting of the precursor to yeast cytochrome c oxidase subunit Va.

We have continued our studies on the import pathway of the precursor to yeast cytochrome c oxidase subunit Va (pVa), a mitochondrial inner membrane protein. Previous work on this precursor demonstrated that import of pVa is unusually efficient, and that inner membrane localization is directed by a membrane-spanning domain in the COOH-terminal third of the protein. Here we report the results of studies aimed at analyzing the intramitochondrial sorting of pVa, as well as the role played by ancillary factors in import and localization of the precursor. We found that pVa was efficiently imported and correctly sorted in mitochondria prepared from yeast strains defective in the function of either mitochondrial heat shock protein (hsp)60 or hsp70. Under identical conditions the import and sorting of another mitochondrial protein, the precursor to the beta subunit of the F1 ATPase, was completely defective. Consistent with previous results demonstrating that the subunit Va precursor is loosely folded, we found that pVa could be efficiently imported into mitochondria after translation in wheat germ extracts. This results suggests that normal levels of extramitochondrial hsp70 are also not required for import of the protein. The results of this study enhance our understanding of the mechanism by which pVa is routed to the mitochondrial inner membrane. They suggest that while the NH2 terminus of pVa is exposed to the matrix and processed by the matrix metalloprotease, the protein remains anchored to the inner membrane before being assembled into a functional holoenzyme complex.

Cell Compartmentation↗
PubMed1992

Use of genetic polymorphisms detected by the random-amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) for differentiation and identification of Aedes aegypti subspecies and populations.

Amplification of random regions of genomic DNA using 10-base primers in the random-amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was used to differentiate and identify mosquito populations based on genetic variation. Genomic DNA was extracted from individual mosquitoes from 11 geographic populations of Aedes aegypti and amplified in PCR reactions using single primers of arbitrary nucleotide sequence. Discriminant analysis of the population frequencies of RAPD fragments produced using three different primers allowed accurate discrimination between the geographic populations in 89% of individuals and between subspecies (Ae. aegypti aegypti versus Ae. aegypti formosus) in 100% of mosquitoes tested. The genetic relatedness of the populations was estimated using three different statistical methods, and unknown populations were correctly classified in a blind test. These results indicate that the RAPD-PCR technique will be useful in studies of arthropod molecular taxonomy and in epidemiologic studies of the relatedness of geographic populations and vector movement.

Aedes↗
PubMed1992

First record of breeding populations of Aedes albopictus in continental Africa: implications for arboviral transmission.

Eggs of Aedes albopictus were collected in oviposition cups from 3 forested areas of Delta State in south-central Nigeria during September 1991 as part of a post-yellow fever outbreak investigation. These eggs were shipped to the Centers for Disease Control in Colorado, where they were reared to the adult stage and identified. This is the first record of breeding populations of Ae. albopictus in continental Africa. Other taxa reared from the same oviposition cups included Ae. aegypti, Ae. apicoargenteus, Ae. africanus, Ae. lilii and Ae. simpsoni subgroup. The introduction and establishment of Ae. albopictus in Africa may have important implications for transmission of indigenous arboviruses.

Aedes↗
PubMed1991

An unusual mitochondrial import pathway for the precursor to yeast cytochrome c oxidase subunit Va.

We have studied the import of the precursor to yeast cytochrome c oxidase subunit Va, a protein of the mitochondrial inner membrane. Like the majority of mitochondrial precursor proteins studied thus far, import of presubunit Va was dependent upon both a membrane potential (delta psi) and the hydrolysis of ATP. However, the levels of ATP necessary for the import of presubunit Va were significantly lower than those required for the import of a different mitochondrial precursor protein, the beta subunit of the F1-ATPase. The rate of import of presubunit Va was found to be unaffected by temperature over the range 0 to 30 degrees C, and was not facilitated by prior denaturation of the protein. These results, in conjunction with those of an earlier study demonstrating that presubunit Va could be efficiently targeted to mitochondria with minimal presequences, suggest that the subunit Va precursor normally exists in a loosely folded conformation. Presubunit Va could also be imported into mitochondria that had been pretreated with high concentrations of trypsin or proteinase K (1 mg/ml and 200 micrograms/ml, respectively). Furthermore, the rate of import into trypsin-treated mitochondria, at both 0 and 30 degrees C, was identical to that observed with the untreated organelles. Thus, import of presubunit Va is not dependent upon the function of a protease-sensitive surface receptor. When taken together, the results of this study suggest that presubunit Va follows an unusual import pathway. While this pathway uses several well-established translocation steps, in its entirety it is distinct from either the receptor-independent pathway used by apocytochrome c, or the more general pathway used by a majority of mitochondrial precursor proteins.

Adenosine Triphosphate↗
PubMed1991

A single amino acid change in the E2 glycoprotein of Venezuelan equine encephalitis virus affects replication and dissemination in Aedes aegypti mosquitoes.

Four monoclonal antibody-resistant variants (MARVs) of Venezuelan equine encephalitis (VEE) virus were used to study mosquito-virus interactions. In vitro experiments using an Aedes albopictus cell line, C6/36, demonstrated that an amino acid change in the glycoprotein E2h epitope (MARV 1A3B-7) decreased virus growth when compared with the wild-type, Trinidad donkey virus, and its vaccine derivative, TC-83. The MARVs replicated as efficiently as the parent virus when inoculated into Aedes aegypti mosquitoes, but MARV 1A3B-7 was restricted in its ability to infect and disseminate from the midgut following oral infection. These results demonstrate that a single amino acid change in the E2 glycoprotein can affect the ability of VEE virus to replicate and disseminate in Ae. aegypti mosquitoes.

Aedes↗
PubMed1991

Genetic selection of a flavivirus-refractory strain of the yellow fever mosquito Aedes aegypti.

Two inbred (isofemale) Aedes aegypti mosquito lines were derived that manifested a resistant or susceptible phenotype following ingestion of yellow fever virus; lack of virus movement from the midgut defined the resistant phenotype. Other flaviviruses, including dengue 1-4, Uganda S, and Zika, viruses behaved in a similar fashion in the two mosquito lines. Crosses between the two lines produced progeny that were of intermediate susceptibility, indicating codominance; F2 backcrosses to the parents yielded results consistent with a major controlling genetic locus and provide evidence of a second locus capable of modulating the phenotype of the major gene. The rapid selection necessary to fix the susceptible and refractory phenotypes support the hypothesis of a single major controlling locus. Viral movement across the midgut is likely to be governed by a single major gene and modifying minor genes or a group of closely linked genes. These inbred mosquito lines will be useful in discovering the molecular basis for flavivirus resistance in Ae. aegypti.

Aedes↗
PubMed1991

Atrial natriuretic factor during hypoxia and mild exercise.

The effect of hypoxia on plasma atrial natriuretic factor (ANF), plasma renin activity (PRA), and plasma aldosterone concentration (PAC) was evaluated during 2 h of treadmill exercise at 2 km/h, 0 grade at sea level. Six male subjects exercised on 2 separate days during normoxia (21% O2) and hypoxia (13.3 +/- 0.3% O2). No significant changes in ANF or PRA occurred during either normoxic or hypoxic exercise. However, PAC fell significantly during normoxic exercise (17.5 +/- 3.6 vs. 12.7 +/- 2.6 ng/dl, p less than 0.05) but not during hypoxic exercise. Serum potassium concentration fell during hypoxic exercise (5.0 +/- 0.1 vs. 4.4 +/- 0.1 mmol/l, p less than 0.05) along with bicarbonate (27.8 +/- 0.7 vs. 25.8 +/- 0.6 mmol/l, p less than 0.01). Between normoxic and hypoxic studies there was a significantly higher heart rate during hypoxic exercise (78 +/- 5 vs. 90 +/- 6 b/min, p less than 0.01). The major conclusion of this study is that hypoxia resulting in arterial oxygen saturations of 81 +/- 0.7% does not affect plasma atrial natriuretic factor levels during mild exercise in normal male subjects.

Adult↗
PubMed1990

Identification of monoclonal antibodies that distinguish between 17D-204 and other strains of yellow fever virus.

Eight monoclonal antibodies (MAbs) prepared against the flaviviruses Saint Louis encephalitis, dengue 2 and dengue 3 viruses all recognized epitopes on the envelope protein of the prototype flavivirus, yellow fever (YF) virus. Three of these MAbs with flavivirus group-common specificity and two MAbs with a flavivirus-subgroup specificity were found to distinguish wild-type YF viruses from YF 17D-204 vaccine virus, but not from the closely related 17DD vaccine virus, nor from the French neurotropic vaccine virus. This pattern of reactivity was seen only with viruses grown in Aedes albopictus C6/36 cells and not with viruses grown in vertebrate cells (SW13 and Vero cells), where all five MAbs recognized epitopes on both wild-type and 17D-204 viruses. Examination of adult A. aegypti mosquitoes infected with the same YF viruses as above gave a different pattern of results to those in C6/36 cells. Thus, epitope expression differs between mammalian and arthropod cells and between arthropod cells in vitro and in vivo. Neutralization tests showed that all five MAbs would neutralize wild-type Asibi virus grown in SW13 cells, but not Asibi virus grown in C6/36 cells, nor 17D-204 vaccine virus grown in either cell type. Therefore, it is concluded that when YF virus is grown in mosquito cells, wild-type virus is antigenically and biologically distinct from the 17D-204 vaccine virus.

Adenocarcinoma↗
PubMed1990

Partial nucleotide sequence of South American yellow fever virus strain 1899/81: structural proteins and NS1.

We have partially cloned and sequenced the genome of a Peruvian yellow fever virus isolate (1899/81) and compared the nucleotide and deduced amino acid sequences of this strain with the previously published sequence of the West African yellow fever virus strain Asibi. In the 3594 base region sequenced, which contains the structural genes (C, M, E), all but the 72 3'-terminal nucleotides of the NS1 gene and 108 nucleotides of the 5' non-coding region, 515 nucleotide substitutions were detected. Nucleotide divergence was lowest in the 5' non-coding region, 2.8%, compared with an average rate of 14.7% in the coding regions. Over 91% of the 512 nucleotide changes in the coding region were silent; 44 amino acid substitutions resulted. The capsid protein was the least conserved, whereas the M protein was the most highly conserved (6.7% and 1.3% divergence, respectively). The envelope protein had 18 amino acid changes (3.7% divergence), one of which created an additional site for potential glycosylation of the 1899/81 virus. NS1 protein divergence (3.9%) was similar to that seen in the E protein. Of the 44 amino acid substitutions found, 34 (77%) were conservative. The highest number of nonconservative differences occurred in the envelope glycoprotein. These changes may significantly affect the antigenic and biological functions of the viruses.

Amino Acid Sequence↗
PubMed1990

Removal of a hydrophobic domain within the mature portion of a mitochondrial inner membrane protein causes its mislocalization to the matrix.

We have examined the import and intramitochondrial localization of the precursor to yeast cytochrome c oxidase subunit Va, a protein of the mitochondrial inner membrane. The results of studies on the import of subunit Va derivatives carrying altered presequences suggest that the uptake of this protein is highly efficient. We found that a presequence of only 5 amino acids (Met-Leu-Ser-Leu-Arg) could direct the import and localization of subunit Va with wild-type efficiency, as judged by several different assays. We also found that subunit Va could be effectively targeted to the mitochondrial inner membrane with a heterologous presequence that failed to direct import of its cognate protein. The results presented here confirmed those of an earlier study and showed clearly that the information required to "sort" subunit Va to the inner membrane resides in the mature protein sequence, not within the presequence per se. We present additional evidence that the aforementioned sorting information is contained, at least in part, in a hydrophobic stretch of 22 amino acids residing within the C-terminal third of the protein. Removal of this domain caused subunit Va to be mislocalized to the mitochondrial matrix.

Amino Acid Sequence↗
PubMed1990

Vector competence of Aedes albopictus for a newly recognized Bunyavirus from mosquitoes collected in Potosi, Missouri.

The vector competence of a Kentucky strain of Aedes albopictus was assessed for a newly recognized Bunyavirus isolated from Ae. albopictus collected in Potosi, Missouri. Females are susceptible to peroral infection and 44.7% became infected after ingesting about 15 Vero cell plaque-forming units (PFU) of virus. Virus replicated and reached average titers of 10(5.4)-10(6.0) PFU/mosquito by day 7 postfeeding. Fourteen (40%) of 35 females tested in an in vitro virus transmission experiment were infected, and 3 (21.4%) of the infected females transmitted virus. There was no evidence of vertical transmission among 1,196 progeny of a group of mothers exposed to infection perorally or among 6,635 progeny of mothers infected by parenteral inoculation. The absence or infrequency of vertical transmission suggests that the virus was not introduced into Missouri via infected Ae. albopictus eggs. Nonetheless, Ae. albopictus is a competent vector of this virus and we provide the first experimental evidence for incriminating Ae. albopictus as a vector in a natural arbovirus transmission cycle in the United States.

Aedes↗
PubMed1990

Vertical transmission of dengue viruses by strains of Aedes albopictus recently introduced into Brazil.

Three strains of Aedes albopictus from Brazil were examined for their ability to vertically transmit dengue 1 (DEN-1) and dengue 4 (DEN-4) viruses. Parental females were uniformly infected by parenteral inoculation of virus, and 8,121 F1 progeny from DEN-1 and DEN-4 infected mothers were pooled in lots of approximately 50 and tested for virus. Seven of 60 pools were positive for DEN-1 virus, and 1 of 121 pools was positive for DEN-4 virus. In DEN-1 assays, the minimum infection rate (MIR) for larvae (2 pools tested) was 1:84. Among positive cohorts of adults, pooled by sex and by geographic strain of mosquito, the MIR ranged from 1:193 to 1:626 for males and from 1:187 to 1:311 for females. Only a single pool of adult females was positive for DEN-4 virus (MIR 1:1022 for an adult female cohort from Santa Teresa). These results indicate that Brazilian Ae. albopictus have the potential to play a role in the maintenance of dengue viruses in nature.

Aedes↗
PubMed1989

Detection of yellow fever viral RNA by nucleic acid hybridization and viral antigen by immunocytochemistry in fixed human liver.

Histopathologic examination of liver from patients with yellow fever is often not diagnostic. We therefore compared 2 virus-specific assays applicable to fixed liver, in situ nucleic acid hybridization and an immunocytochemical [alkaline phosphatase-antialkaline phosphatase (APAAP)] technique. Yellow fever structural gene sequences were detected by use of 35S-labeled negative-sense RNA probe (but not by immunocytochemistry) in 11 of 17 livers from children with fatal illness during the 1965 epidemic in Senegal. These fixed liver samples had been stored at ambient temperatures for 23 years. Both techniques were diagnostic on tissues collected 15-37 months before testing. Immunocytochemistry is a practical procedure for rapid specific diagnosis of liver stored for months, whereas RNA-RNA hybridization is a sensitive technique which can be applied to material stored for years.

Adolescent↗
PubMed1989

Co-circulation of multiple Colorado tick fever virus genotypes.

Colorado tick fever (CTF) virus, family Reoviridae, genus Orbivirus, contains 12 genes distinguishable by polyacrylamide gel electrophoresis (PAGE). Multiple genotypes of CTF virus were isolated at 3 field sites in Colorado in 1985. Five genotypes were found at Campos Cabin, 2 at Drake, and 6 at Rocky Mountain National Park. Virus isolations were made in 1985 from 6 patients with CTF. These isolates were distinct from each other and the field isolates. Although the CTF isolates were different by PAGE profile, the majority of the 12 genes were highly conserved among the 1985 isolates and a Florio isolate (FMA). Only genes 4 and 6 were variant among the 1985 CTF isolates and FMA, and no unique genes were identified. In 1986, a follow-up field survey was done at the Campos Cabin site. Of the 3 CTF PAGE genotypes obtained, 2 exhibited PAGE profiles which were different from the 1985 isolates. One isolate may have resulted from the reassortment of genes from 2 of the isolates circulating at Campos Cabin in 1985.

Animals↗
PubMed1989

Pyogenic vertebral osteomyelitis after transcolonic gunshot wound.

Recently, pyogenic vertebral osteomyelitis has been documented following transcolonic passage of bullets lodging in or near the vertebral column. It is becoming more recognized that foreign body missiles are not as sterile as was once thought--especially after passage through the gastrointestinal tract--and that this condition contributes to the incidence of infection in such bone injuries. In this article the problem of deciding on and implementing proper initial treatment and management of vertebral injuries caused by foreign body missiles is illustrated and discussed.

Adult↗
PubMed1989

Epidemic yellow fever caused by an incompetent mosquito vector.

Arbovirus epidemics in a geographic region are believed to depend on the presence of susceptible or "competent" arthropod vectors. We demonstrate that an urban, Aedes aegypti-borne, epidemic of yellow fever occurred in 1987 although the mosquito vector was relatively resistant to infection and transmitted the virus inefficiently. Twenty-six percent of the experimental mosquitoes from the epidemic area that ingested yellow fever virus became infected and only 7% of these transmitted the virus. In contrast, 80% of an exotic susceptible strain of Ae. aegypti became infected and 43% were able to transmit. We also show that no other potential vectors were active during the epidemic and that the local Ae. aegypti were present in extremely large numbers. These results document, for the first time, that, in the presence of high population density an incompetent mosquito vector can initiate and maintain virus transmission resulting in an epidemic.

Adult↗