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Biomedical subjects

R C Rose

Publications and source records attributed to R C Rose.

At least 19 recordsLinked to original sources
PubMed1999

Nuclear import of HPV11 L1 capsid protein is mediated by karyopherin alpha2beta1 heterodimers.

L1 major capsid proteins of human papillomaviruses (HPVs) enter the nuclei of host cells at two times during the viral life cycle: 1) after infection and 2) later during the productive phase, when they assemble the replicated HPV genomic DNA into infectious virions. L1 proteins are stable in two oligomeric configurations: as homopentameric capsomers, and as capsids composed of 72 capsomers. We found that intact L1 capsids of HPV type 11 cannot enter the nucleus, suggesting that capsid disassembly may be required for HPV11 L1 nuclear import. We established that HPV11 L1 is imported in a receptor-mediated manner into the nuclei of digitonin-permeabilized HeLa cells. HPV11 L1 docked at the nuclear pore complexes via karyopherin alpha2beta1 heterodimers. Anti-karyopherin-beta1 and anti-karyopherin alpha2 antibodies specifically inhibited nuclear import of HPV11 L1. Moreover, nuclear import of HPV11 L1 could be reconstituted using karyopherin alpha2, beta1, RanGDP and p10. In agreement with the docking and import data, we found that HPV11 L1 binds to karyopherin alpha2 and that this interaction is inhibited by a peptide representing the classical nuclear localization signal of SV40 T antigen. These results strongly suggest that HPV11 L1 enters the nucleus of the infected host cell via the karyopherin alpha2beta1 pathway.

Amino Acid Sequence↗
PubMed1999

Oral vaccination of mice with human papillomavirus virus-like particles induces systemic virus-neutralizing antibodies.

To assess whether oral vaccination against human papillomavirus (HPV) may be feasible, we administered HPV virus-like particles (VLPs) to mice by gavage. Enzyme-linked immunosorbent assay (ELISA) results indicated that serum anti-VLP immunoglobulin G (IgG) and IgA antibodies were induced after oral vaccination, and these responses demonstrated antigenic specificities that were conformationally dependent and restricted according to HPV genotype. Importantly, orally induced postimmune sera were found to neutralize HPV-11 virions in vitro. These results indicated that the VLPs were antigenically stable in the environment of the gastrointestinal tract and were able to engage in potentially useful immune system interactions. These findings support the concept of oral vaccination against anogenital HPV disease, and suggest the possibility that this may be a useful approach to the immunization of large populations against cervical cancer and other HPV associated diseases.

Administration, Oral↗
PubMed1999

Glutathione-dependent ascorbate recycling activity of rat serum albumin.

An efficient regeneration of vitamin C (ascorbate) from its oxidized byproduct, dehydroascorbate (DHAA), is necessary to maintain sufficient tissue levels of the reduced form of the vitamin. Additionally, the recycling may be more significant in mammals, such as guinea pigs and humans, who have lost the ability to synthesize ascorbate de novo, than it is in most other mammals who have retained the ability to synthesize the vitamin from glucose. Both a chemical and an enzymatic reduction of DHAA to ascorbate have been proposed. Several reports have appeared in which proteins, including thioltransferase, protein disulfide isomerase, and 3-alpha-hydroxysteroid dehydrogenase, characterized for other activities have been identified as having DHAA reductase activity in vitro. Whether these previously characterized proteins catalyze the reduction of DHAA in vivo is unclear. In the present study, a 66 kD protein was purified strictly on the basis of its DHAA-reductase activity and was identified as rat serum albumin. The protein was further characterized and results support the suggestion that serum albumin acts as an antioxidant and exerts a significant glutathione-dependent DHAA-reductase activity that may be important in the physiologic recycling of ascorbic acid.

Amino Acid Sequence↗
PubMed1998

Water soluble antioxidants in mammalian aqueous humor: interaction with UV B and hydrogen peroxide.

HPLC/electrochemical detection was used to identify five major low MW water soluble electrochemically active molecules from the aqueous humor of three species of mammals: New Zealand White rabbits and humans (diurnal) and Sprague-Dawley rats (nocturnal). These molecules are L-cysteine (CYS), L-ascorbic acid (AA), glutathione (GSH), uric acid (UA) and L-tyrosine (TYR); all of these molecules have known antioxidant properties. Nocturnal rat aqueous humor is concentrated in two thiols: GSH (125 microM; n = 24 pooled eyes) and CYS (63 microM), in contradistinction to diurnal species which have high concentrations of AA. No deterioration of any of these antioxidants occurs in a synthetic aqueous humor mixture irradiated with a physiologically relevant spectral UV B dose of 30 mJ/cm2/h (5.5 UV equivalent sunlight hours). The same result occurred with addition of the endogenous aqueous humor UV B photosensitizer L-tryptophan. In a second set of experiments, human synthetic aqueous humor was subjected to hydrogen peroxide induced oxidant stress. The decay of antioxidants was CYS > GSH > AA > UA > TYR. The second highest concentrated antioxidant in human aqueous humor is TYR. Yet TYR failed to protect AA against H2O2-induced free radical damage in a synthetic aqueous humor model system (P = 0.10; ANOVA). The existence of multiple electrochemically active constituents and their thermodynamic interactions must be recognized when choosing animal models to evaluate human aqueous humor antioxidant defense.

Aged↗
PubMed1998

Vitamin supplementation and breast cancer: is homeostasis a factor?

Recent observations by several research groups on many thousands of women have yielded the disappointing view that mega-dose vitamin supplementation does not provide significant protection against breast cancer. This is a review of the pertinent literature with a goal of identifying testable hypotheses that might explain the epidemiology and be helpful in designing subsequent evaluations. In one hypothesis presented, the vitamin content of peripheral cells that protect breast epithelium is not markedly affected by supplementation. In the second hypothesis the metabolic status (redox state) of epithelial cells is more important than the absolute level (reduced plus oxidized) of each antioxidant. In either case, extremes in diet fail to alter inherent homeostatic mechanisms.

Breast Neoplasms↗
PubMed1998

Tear fluid content of electrochemically active components including water soluble antioxidants.

PURPOSE: To describe the content of water-soluble antioxidants in tear fluid. METHODS: We collected tear fluid from healthy subjects, either into borosilicate glass tubing or by absorption onto Schirmer strips. High pressure liquid chromatography with electrochemical detection provided data on several components in a single assay. The system was sufficiently sensitive to provide reliable values for components present in tear fluid collected at normal (basal) or stimulated rates of secretion. RESULTS: Tear fluid at basal secretion was found to contain four compounds often considered to function in biology as antioxidants. Ascorbic acid (AA) is found at 665 microM, tyrosine at 45 microM and glutathione (GSH) at 107 microM. Cysteine (48 microM) and uric acid (328 microM) are reported for the first time; the latter is somewhat controversial as a physiologically active antioxidant. One peak on the chromatogram was consistently present but has not been identified. During stimulation of flow by brief inhalation of ammonium hydroxide fumes, the concentration of each compound was lower. CONCLUSION: This work demonstrates the presence of electro-chemically active components that might function as antioxidants at the anterior surface of the cornea against potential damage from radiation, oxygen toxicity, abrasion and environmental chemicals.

Adult↗
PubMed1998

In vitro infection and type-restricted antibody-mediated neutralization of authentic human papillomavirus type 16.

Human papillomavirus type 16 (HPV-16) is strongly associated with the development of cervical cancer. Studies of model systems with animal papillomaviruses have demonstrated the importance of neutralizing antibodies in preventing papillomavirus-associated disease. The assessment of neutralizing antibody responses against HPV-16, previously hampered by the lack of a viral source, was enabled by the recent propagation of an HPV-16 stock in xenografted severe combined immunodeficiency (SCID) mice. HPV-16 infection of an immortalized human keratinocyte cell line was demonstrated by detection of an HPV-16-specific spliced mRNA amplified by reverse transcriptase PCR. Infection was blocked by preincubation of the virus with antiserum generated against HPV-16 virus-like particles (VLPs) composed of the major capsid protein, L1. To examine potential cross-neutralizing activity among the different genital HPV types, rabbit antisera to L1 VLPs corresponding to HPV-6, -11, -18, -31, -33, -35, -39, and -45 were assayed for the ability to block the HPV-16 infection of cultured cells. Antiserum raised against HPV-33 L1 VLPs was the only heterologous antiserum which inhibited HPV-16 infection. Thus, a neutralization assay for HPV-16 may help to characterize the components required to compose a broadly efficacious genital HPV vaccine.

Animals↗
PubMed1998

Isolation and propagation of human papillomavirus type 16 in human xenografts implanted in the severe combined immunodeficiency mouse.

We report the isolation and propagation of human papillomavirus type 16, the main agent of cervical cancer, using human foreskin fragments implanted in severe combined immunodeficiency mice. The infection produced viral particles, and with each passage of the virus it caused lesions identical to intraepithelial neoplasia, the precursor to carcinoma.

Animals↗
PubMed1998

Human papillomavirus type 11 recombinant L1 capsomeres induce virus-neutralizing antibodies.

The human papillomavirus type 11 (HPV-11) L1 major capsid protein can be trypsinized to generate recombinant capsomeres that retain HPV genotype-restricted capsid antigenicity (M. Li, T. P. Cripe, P. A. Estes, M. K. Lyon, R. C. Rose, and R. L. Garcea, J. Virol. 71:2988-2995, 1997). In the present study, HPV-11 virion-neutralizing monoclonal antibodies H11.F1 and H11.H3, previously characterized as recognizing two distinct HPV-11 capsid-neutralizing antigenic domains (S. W. Ludmerer, D. Benincasa, and G. E. Mark III, J. Virol. 70:4791-4794, 1996), were each found to be highly immunoreactive with trypsin-generated capsomeres in an enzyme-linked immunosorbent assay (ELISA). Capsomeres were used to generate high-titer polyclonal immune sera that demonstrated HPV genotype-restricted reactivity by ELISA. The capsomere antisera were then tested in an in vitro infectivity assay and found to neutralize HPV-11 virions. In this assay, HPV-11 capsomere polyclonal antisera exhibited neutralization titers (10(-5) to 10(-6)) comparable to those obtained with a virion-neutralizing antiserum raised previously against intact HPV-11 VLPs (R. C. Rose, R. C. Reichman, and W. Bonnez, J. Gen. Virol. 75:2075-2079, 1994). These results indicate that highly immunogenic, genotype-restricted HPV capsid-neutralizing antigenic domains are contained entirely within capsomeres. Thus, capsomeres may be viable vaccine candidates for the prevention of HPV disease.

Animals↗
PubMed1998

Ocular oxidants and antioxidant protection.

Oxidative damage and antioxidant protection in ocular tissues has not been reviewed recently. Metabolism in the eye is of increasing interest because the organ is highly susceptible to damage by sunlight, oxygen, various chemicals, and pollutants. Interest is expected to increase because of an aging Western world population and a continued depletion of stratospheric ozone. Hydrogen peroxide is discussed because it is both a byproduct and a source of free radical reactions and is normally present in the aqueous humor. The metabolism of reactive oxygen species by enzymes, nutrients, pigments, and low molecular weight scavengers is evaluated. Ascorbic acid, because of its high concentration in the eye, is thought to be a primary substrate in ocular protection; progress in determining the mechanisms by which it is recycled and maintained in the useful, reduced state is discussed. Recent information is included about antioxidants not previously known to be present in the eye, and some importance is placed on the properties of the vitreous humor and tear fluid because of the previous lack of emphasis on these.

Animals↗
PubMed1997

Age distribution of antibodies to human papillomavirus in children, women with cervical intraepithelial neoplasia and blood donors from South Africa.

Sera from 95 women with cervical intraepithelial neoplasia (CIN), 95 age-matched female blood donors, and 155 children aged between 1 and 12 years were tested by enzyme-linked immunosorbent assay (ELISA) for levels of serum IgG to three human papillomavirus (HPV) peptides (HPV-16 E2 [E2-16], HPV-18 E2 (E2-18], HPV-16 L1 [L1-16]), as well as HPV-16 virus-like particles (VLP-16) and bovine papillomavirus type 1 virus-like particles (BPV-VLP). In the adult group antibodies to E2-16 and VLP-16 were significantly associated with CIN when compared to the blood donor controls (P = .039 and P = .002, respectively). In women with CIN there was an increase in seropositivity to E2-16 and a decrease in seropositivity to VLP-16 with age. Antibodies to HPV-16 E2 could therefore be an important marker of CIN in women over 40 years of age, whereas antibodies to VLP-16 could be a marker for CIN in younger women. There was no correlation with CIN and antibodies to E2-18, L1-16, and BPV-VLP. In the children's sera antibodies were detected to E2-16 (44.5%), E2-18 (18.7%), L1-16 (20%), VLP-16 (4.5%), and BPV-VLP (5.1%). Between the ages of 3 and 12 years the prevalence of antibodies to E2-16 decreased with age. The presence of antibodies to HPV-16 in young children indicated infection with either HPV-16 or a related virus. HPV DNA isolation from children could help resolve this question.

Adult↗
PubMed1997

Properties of electrochemically active components in mammalian vitreous humor.

Antioxidants are considered to have important roles in protecting the eye against radiation, oxygen toxicity and other threats. Several water-soluble electrochemically-active compounds that might function as antioxidants in vitreous humor of rabbit, rat and bovine eyes were identified and quantitated. In that the rat is a nocturnal animal species and the other two are diurnal, the results allow a tentative evaluation on the comparative physiology of light-dark behavior patterns that influence this ocular fluid. All analyses were performed by HPLC with electrochemical detection. The utility of this approach is that components of vitreous humor can be identified by their retention time and by their characteristic response when the voltages of the two sensing electrodes are progressively brought together. Four compounds regarded as functional antioxidants in the animal kingdom were identified and quantitated. Other antioxidants known to be present in several biological fluids are not present at detectable or significant levels in vitreous humor; metabolites or precursors of these might be present in trace quantities.

Animals↗
PubMed1997

Serological and T-helper cell responses to human papillomavirus type 16 L1 in women with cervical dysplasia or cervical carcinoma and in healthy controls.

In a cross-sectional study we have investigated serological and T-helper (Th) cell responses to human papillomavirus type 16 (HPV-16) L1 in women with HPV-16 related diseases and related them to cervical histology and HPV DNA status. Using a virus-like particle (VLP) based ELISA to detect antibodies to the HPV-16 L1 capsid protein, 45% (33/73) of women with cervical dysplasia, 40% (2/5) of women with cervical cancer, 36% (4/11) of healthy adult female controls and 6% (2/35) of healthy children were found to be seropositive. Amongst women with cervical dysplasia, the highest levels of seropositivity were found in those who were HPV-16 DNA positive (60%, 15/25) or positive for any of the "high-risk' HPV types, 16/18/33 (58%, 18/31), when compared with those with HPV type "X' (25%, 5/20) or with healthy children (6%, 2/35; P < 0.05 for all comparisons). There was a trend for women with cervical dysplasia to show an increased level of seropositivity with increasing grade of lesion. There was no direct correlation found between seropositivity and Th cell responses in all groups studied. However, a combined analysis of each individual's Th and B cell responses suggests that a Th1 pattern of response is predominant amongst healthy adult controls (80% of responders) but reduced in women with cervical dysplasia (55% of responders). A trend towards a decrease in Th1 type responses was also noted with increasing grade of dysplastic lesion. These findings provide further evidence for the importance of the Th response in the control of genital HPV infections.

Adult↗
PubMed1997

Expression of the human papillomavirus type 11 L1 capsid protein in Escherichia coli: characterization of protein domains involved in DNA binding and capsid assembly.

The L1 major capsid protein of human papillomavirus type 11 (HPV-11) was expressed in Escherichia coli, and the soluble recombinant protein was purified to near homogeneity. The recombinant L1 protein bound DNA as determined by the Southwestern assay method, and recombinant mutant L1 proteins localized the DNA-binding domain to the carboxy-terminal 11 amino acids of L1. Trypsin digestion of the full-length L1 protein yielded a discrete 42-kDa product (trpL1), determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, resulting from cleavage at R415, 86 amino acids from the L1 carboxy terminus. Sucrose gradient sedimentation analysis demonstrated that trpL1 sedimented at 11S, while L1 proteins with amino-terminal deletions of 29 and 61 residues sedimented at 4S. Electron microscopy showed that the full-length L1 protein appeared as pentameric capsomeres which self-assembled into capsid-like particles. The trpL1 protein also had a pentameric morphology but was unable to assemble further. In an enzyme-linked immunosorbent assay, the trpL1 and L1 capsids reacted indistinguishably from virus-like particles purified after expression of HPV-11 L1 in insect cells. The carboxy terminus of L1 therefore constitutes the interpentamer linker arm responsible for HPV-11 capsid formation, much like the carboxy-terminal domain of the polyomavirus VP1 protein. The trypsin susceptibility of HPV-11 L1 capsids suggests a possible mechanism for virion disassembly.

Amino Acid Sequence↗
PubMed1996

An apparatus for ultraviolet B irradiation of small volumes of biological fluid under controlled oxygen tension.

BACKGROUND: The design, construction, and calibration of a unique apparatus for ultraviolet (UV) B irradiation of small volumes of biological fluid under temperature-controlled open atmospheric or closed cell conditions of defined Po2 is described. METHODS: The apparatus consists of a custom designed UV transmissible quartz silica 3 port spectrophotometric cuvette coupled to a deuterium UV B source by UV transmissible fiber optics. Fluid movement, temperature regulation, dosimetry, and miniaturized biosensors for oxygen, pH, and temperature are described along with a preliminary experiment. RESULTS: The apparatus produces a known photochemical effect using an endogenous aqueous humor UV B photosensitizer-L tryptophan. UV B radiation is contained, thus protecting laboratory personnel. The system provides a continuous spectral UV B dose of 30 mJ/cm2/hr, which is equivalent to approximately 5.5 UV sunlight hours reaching an aliquot of mammalian aqueous humor. CONCLUSIONS: This biophysical apparatus can be used to investigate antioxidant protective systems in ocular fluid (aqueous/vitreous humors and tears), and is also applicable to evaluating the interaction of radiation with ocular and nonocular tissue cell homogenate.

Animals↗
PubMed1996

The potential role of speech production models in automatic speech recognition.

This paper investigates the issues that are associated with applying speech production models to automatic speech recognition (ASR). Here the applicability of articulatory representations to ASR is considered independently of the role of articulatory representations in speech perception. While the question of whether it is necessary or even possible for human listeners to recover the state of the articulators during the process of perceiving speech is an important one, it is not considered here. Hence, the authors refrain from posing completely new paradigms for ASR which more closely parallel the relationship between speech production and human speech understanding. Instead, work aimed at integrating speech production models into existing ASR formalisms is described.

Humans↗
PubMed1996

Intestinal absorption of water-soluble vitamins.

Vitamin utility is a scientific/medical topic that appears to be pursued as ardently by the lay public as by scientists and medical practitioners. A group of epidemiologists evaluates the effects on health of vitamin intake in the natural diet and by supplementation. The role in ocular disease, cancer of the breast or colon, and cardiovascular disease are a few of the concerns. The results and recommendations concerning dietary vitamin intake will likely continue to change. However, the processes by which dietary vitamins are delivered from intestinal chyme to the blood are more certain. The concept of homeostasis might apply to various of the vitamins as it does to minerals, water, etc. This review will discuss some common methods used to study vitamin absorption and the proposed mechanisms of absorption, and will conclude with a section about dietary regulation.

Animals↗