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Biomedical subjects

C A Wilson

Publications and source records attributed to C A Wilson.

At least 109 records · Page 6Linked to original sources
PubMed1994

Quantitative micro P30 and reverse transcriptase assays for Moloney murine leukemia virus.

An anti-P30 immunohistochemical and a reverse transcriptase assay for Moloney murine leukemia virus (MoMLV) are adapted to 96-well plates. The assay results are shown to be directly proportional to the number of infectious particles, and can therefore be used to estimate the infective titers of a virus preparation. The micro P30 assay yields a direct estimate of infectious centers, and the reverse transcriptase assay quantitates progeny from a single cycle of replication. The semi-automated nature of these assays is well suited to the analysis of a large number of samples and therefore permits the examination of the efficiency of the process of retroviral/MoMLV infection under varied times or conditions.

3T3 Cells↗
PubMed1994

Visualization of subtle contrast-related intensity changes using temporal correlation.

Contrast-enhanced magnetic resonance imaging (MRI) is a promising method for investigating the breakdown of the blood-retinal barrier (BRB). However, subtle intensity changes due to low concentrations of contrast agent can be difficult to detect without observer bias. In this study, we developed a temporal correlation method for detecting these subtle signal intensity changes. The method was evaluated in eyes with chemically induced retinal lesions of known size. A time series of MRI data were collected following i.v. administration of different doses of gadolinium-diethylaminetriaminepentaacetic acid (0.05, 0.1, 0.5 mmol/kg). These time course images were analyzed by temporal correlation to a reference enhancement curve. The reference curve was generated based on a validated theoretical enhancement curve. The temporal correlation method detected signal intensity changes in cases where the changes were too subtle to be visible on a postinjection image or a subtraction image (obtained by subtracting the precontrast image from the final image in the time course set). In addition, assessment of leakage was performed by viewing each image in the set with an eight gray-level palette. Areas of leakage identified in this manner corresponded to those identified by temporal correlation, a finding which supports the validity of the temporal correlation method. These results suggest that temporal correlation may be a time-efficient way to screen large numbers of image data sets using an objective, user-independent criterion.

Algorithms↗
PubMed1994

Comparison of cDNAs encoding the gibbon ape leukaemia virus receptor from susceptible and non-susceptible murine cells.

The gibbon ape leukaemia virus (GaLV) family of type C retroviruses consists of five closely related viral isolates, GaLV SF, GaLV SEATO, GaLV Br, GaLV H and simian sarcoma-associated virus. The cDNA encoding the human receptor for GaLV SEATO had previously been isolated. We now demonstrate that all of the above GaLVs can use the human form of the GaLV receptor to infect cells. All murine cells analysed to date have been found to be resistant to infection by GaLVs owing to the absence of a functional GaLV receptor. We have now identified a murine cell line which is unique in its susceptibility to GaLV infection. This cell line was established from a Japanese feral mouse, Mus musculus molossinus. We cloned and sequenced the cDNA for the receptor expressed in these cells and compared it to the cDNA for the GaLV receptor expressed in resistant murine cells such as NIH 3T3 (derived from M. m. musculus) and MDTF (derived from M. dunni tail fibroblasts). The crucial region for GaLV infection (the fourth extracellular domain) from the functional M. m. molossinus GaLV receptor is quite divergent from the same region of the M. m. musculus and M. dunni proteins, but similar to that of the functional human GaLV receptor. These results confirm the importance of the amino acids of this region in GaLV receptor function.

3T3 Cells↗
PubMed1994

Clothing and bedding and its relevance to sudden infant death syndrome: further results from the New Zealand Cot Death Study.

As part of a large nationwide case-control study covering a region with 78% of all births in New Zealand during 1987-90, the clothing and bedding of infants dying of sudden infant death syndrome (SIDS) and that of an appropriate control group were recorded. Cases consisted of 81% (n = 393) of all cases of SIDS in the study area and 88.4% (n = 1592) of 1800 control infants randomly selected from the hospital births and who completed a home interview. Use of a wool 'waterproof' underblanket was associated with a significantly reduced risk of SIDS (adjusted OR 0.44; 95% CI: 0.26-0.73) while sheepskin use was not. Firm tucking was identified as contributing to a reduced risk of SIDS even after adjusting for potentially confounding variables (adjusted OR 0.63, 95% CI: 0.46-0.86). Sixty case infants (15.6% of cases) were found dead with the head covered but there were no equivalent data for controls. Having been found previously completely covered by bedding was equally common in cases and controls (28.8% cases and 30.6% of control infants). Other differences of bedding and clothing between cases and controls were small; mattress characteristics were not studied. The exact methods in which babies are cared for are important and this study suggests that infants are at lower risk of SIDS when firmly tucked in and when sleeping on a 'waterproof' wool underblanket.

Bedding and Linens↗
PubMed1994

Properties of a unique form of the murine amphotropic leukemia virus receptor expressed on hamster cells.

Identification and cloning of the receptors for amphotropic murine leukemia virus (A-MuLV) and gibbon ape leukemia virus (GaLV) have both enabled the determination of the normal function of these virus receptors in cells and initiated experimental examination of how these receptors interact with their respective viruses. GaLV and A-MuLV have distinct host ranges and use different receptors to infect human cells. It was therefore surprising to find that the human GaLV and A-MuLV receptors were not only structurally similar but performed similar cellular functions (B. O'Hara, S. V. Johann, H. P. Klinger, D. G. Blair, H. Rubinson, K. J. Dunn, P. Sass, S. M. Vitek, and T. Robbins, Cell Growth Differ. 1:119-127, 1990; M. van Zeijl, S. V. Johann, E. Closs, J. Cunningham, R. Eddy, T. B. Shows, and B. O'Hara, Proc. Natl. Acad. Sci. USA 91:1168-1172, 1994; M. P. Kavanaugh, D. G. Miller, W. Zhang, W. Law, S. L. Kozak, D. Kabat, and A. D. Miller, Proc. Natl. Acad. Sci. USA 91:7071-7075, 1994; and Z. Olah, C. Lehel, W. B. Anderson, M. V. Eiden, and C. A. Wilson, J. Biol. Chem., in press). We have now determined that the murine retrovirus 10A1 can use both the human GaLV receptor and the human A-MuLV receptor to infect cells. Furthermore, we have cloned and functionally characterized a unique form of the amphotropic receptor homolog expressed in E36 hamster cells. This receptor (EAR) can serve as both a GaLV receptor and an A-MuLV receptor, and it therefore differs from the receptors expressed in human cells, which function exclusively as either GaLV or A-MuLV receptors.

Amino Acid Sequence↗
PubMed1994

Glycosylation-dependent inactivation of the ecotropic murine leukemia virus receptor.

The ecotropic murine leukemia virus (E-MuLV) receptor expressed on Mus dunni tail fibroblast (MDTF) cells is a receptor for all E-MuLVs with the notable of Moloney murine leukemia virus (Mo-MuLV). Substitution of isoleucine for valine at position 214 in the third extracellular region (the putative E-MuLV binding site) of the MDTF receptor molecule allows this molecule to function as a Mo-MuLV receptor (M.V. Eiden, K. Farrell, J. Warsowe, L. A. Mahan, and C. A. Wilson, J. Virol. 67:4056-4061, 1993). We have now determined that treating MDTF cells with tunicamycin, an inhibitor of N-linked glycosylation, also renders them susceptible to Mo-MuLV infection. Two potential N-linked glycosylation sites are present in the third extracellular regions of both the NIH 3T3 and MDTF ecotropic receptors. The glycosylation site at position 229 of the MDTF receptor cDNA was eliminated by substituting a threonine codon for the asparagine codon. Mo-MuLV-resistant human HOS cells, expressing this form of the receptor, are susceptible to Mo-MuLV infection. Thus, our studies suggest that without a glycan moiety at position 229, the valine residue at 214 is no longer restrictive for Mo-MuLV infection. BHK-21 and CHO K1 hamster cells also express glycosylation-inactivated forms of the ecotropic receptor. Sequence analysis of these receptors together with our analysis of MDTF receptor function suggests that a single asparagine-linked glycosylation site is responsible for glycosylation inactivation of these receptors.

Amino Acid Sequence↗
PubMed1994

Classification and localisation of carcinoembryonic antigen (CEA) related antigen expression in normal oesophageal squamous mucosa and squamous carcinoma.

Using a panel of carcinoembryonic antigen (CEA) related antibodies in normal oesophageal squamous mucosa CEA expression is present on suprabasal squames localised to the cell membrane. Immunoblotting shows that this positivity is predominantly due to a glycoprotein of around 180 kDa representing CEA itself. Positivity in squamous carcinomas is confined to cells in foci of squamous differentiation. A shift from membranous localisation to predominant cytoplasmic overexpression is shown between normal and malignant squames using confocal microscopy. The recognition of an adhesive role for CEA and a role in enhancing distant metastases in those tumours expressing CEA highlights the importance of recording CEA expression and changes in subcellular distribution between normal and malignant tissues; CEA expression in oesophageal squamous mucosa has not been well recognised previously and changes in expression may prove of great significance in the spread and dissemination of squamous carcinoma.

Carcinoembryonic Antigen↗
PubMed1994

The physicochemical, immunological and biological properties of rat pituitary and plasma LH.

A comparison was made between the properties of LH derived from female rat pituitary glands and plasma. Samples were collected from adult intact rats 5 h before or at the time of the pro-oestrous preovulatory LH surge; 27-day-old rats untreated or given 5 IU pregnant mares' serum gonadotrophin (PMSG) s.c. on day 25, which induced LH release 54 h later and adult ovariectomized rats untreated or primed with either 5 micrograms oestradiol benzoate s.c. or 5 micrograms oestradiol benzoate followed 48 h later by 0.5 mg progesterone s.c., which induced LH release 4-6 h later. All pituitary LH samples were totally bound to an anionic ion-exchange resin (DE52), while only a small proportion of the plasma LH was bound. Only 0-10% plasma LH obtained from intact, ovariectomized (with and without steroids) and untreated immature rats was bound, while a greater proportion of bound LH (36%) was noted in rats treated with PMSG. Gel filtration indicated only slight differences between pituitary and plasma LH, the former eluting marginally earlier than whole plasma and the unbound and bound plasma forms derived after separation by DE52 resin. Affinity chromatography (Concanavalin A and Glycine maximus) showed that LH from both sources possesses high mannose oligosaccharides and that plasma LH does not bear terminal N-acetyl galactosamine residues, although 20% of the pituitary form does. Plasma obtained from pro-oestrous rats had greater bioactivity than had pituitary LH in stimulating testosterone from Leydig cells and progesterone from granulosa cells in vitro, and inducing ovulation in immature rats in vivo. Leydig cell bioassays for LH in fractions obtained from ion-exchange separation indicate that steroidogenic activity of unbound plasma LH is greater than bound pituitary LH when they were collected at times of enhanced release. When release was inhibited (oestrogen-primed ovariectomized rats or immature rats), the steroidogenic activity of plasma and pituitary LH were similar and an acidic steroidogenic component was present in the plasma that was not recognized immunogenically as LH. In summary, pituitary LH undergoes a conversion on release into the plasma that involves a change in binding characteristics on an ion-exchange resin. In conditions when LH release is enhanced there is an increase in bioactivity of plasma LH owing to modification either by steroids or some other plasma factor(s) that perhaps influence the structure of LH directly or by steroids acting indirectly to alter GnRH release, which then modifies LH structure. These structural changes are minor and probably involve alterations in the glycosyl attachments.

Animals↗
PubMed1994

Detection of intraabdominal abscess by serum lysozyme estimation.

BACKGROUND: The failure of new, innovative, and often expensive tests to show the presence of intraabdominal infection caused us to reexamine the value of an inexpensive and almost forgotten one. Lysozyme is a bacteriolytic enzyme located within the lysosomes of phagocytic cells including leukocytes. METHODS: We measured serum concentrations of lysozyme by a standard turbidimetric method in both a murine model of intraabdominal infection and in trauma patients with intraabdominal abscesses or other acquired infections. RESULTS: In mice with intraabdominal abscess secondary to cecal ligation and puncture (n = 35) serum lysozyme activity increased compared with sham-operated controls (n = 20; p < 0.001). In trauma patients with intraabdominal abscess after injury and surgery (n = 19), there was also an increase in serum lysozyme activity compared with controls (n = 15; p < 0.001) or with patients with lung infection (n = 21; p < 0.001). The increase of serum lysozyme activity occurred before intraabdominal sepsis was clinically apparent. CONCLUSIONS: In this study, serum lysozyme concentration has a high specificity related to the presence of an intraabdominal abscess and is an indirect measure of the sequestration of leukocytes to the site of an established or developing collection of intraabdominal pus. The estimation of serum lysozyme may be an aid to differentiate critically ill patients with a potential occult intraabdominal abscess.

Abdomen↗
PubMed1993

Evaluating treatment efficacy by triangulation.

The project 'Efficacy of Support Groups for Mexican American Widows' was evaluated by triangulating results collected from different points of view. These included changes after one year in (1) measurement scores of standardized instruments, (2) assessment of adjustment to role of widow by the outsider and (3) evaluation by the widow herself. When subjects were categorized by their assignment to experimental or control groups, the scores on standardized instruments including the Beck Depression Inventory (BDI), the Center for Epidemiologic Study of Depression (CES-D), Anxiety State, Life Satisfaction and new scales measuring Somatic symptoms and Emotional symptoms had decreased from time 1 to time 5 for all widows. However, the widows participating in support groups showed significantly improved changes in these scores. The outsiders found the women who participated in the support groups to have a far better change to the role of widow than the control widows. The support group widows themselves described more positive changes in themselves than did control widows.

Adolescent↗
PubMed1993

Immune escape by human immunodeficiency virus type 1 from neutralizing antibodies: evidence for multiple pathways.

Sera from many HIV-1-infected individuals contain broadly reactive, specific neutralizing antibodies. Despite their broad reactivity, variant viruses, resistant to neutralization, can be selected in vitro in the presence of such antisera. We have previously shown that neutralization resistance of an escape mutant with an amino acid substitution in the transmembrane protein (A582T) occurs because of alteration of a conformational epitope that is recognized by neutralizing antibodies directed against the CD4 binding site. In this report we demonstrate that immune escape via a single-amino-acid substitution (A281V) within a conserved region of the envelope glycoprotein gp120 confers neutralization resistance against a broadly reactive neutralizing antiserum from a seropositive individual. We show this alteration affects V3 and additional regions unrelated to V3 or the CD4 binding site. Together with previous studies on escape mutants selected in vitro, our findings suggest that immune-selective pressure can arise by multiple pathways.

Amino Acid Sequence↗
PubMed1993

Characterization of a naturally occurring ecotropic receptor that does not facilitate entry of all ecotropic murine retroviruses.

A fibroblast cell line (MDTF) derived from the feral mouse Mus dunni is resistant to infection by Moloney murine leukemia virus (Mo-MuLV), an ecotropic murine leukemia virus (E-MuLV) (M. R. Lander and S. K. Chattopadadhyay, J. Virol. 52:695-698, 1984). MDTF cells can be infected by other E-MuLVs such as Friend MuLV and Rauscher MuLV, which have been demonstrated to use the same receptor as Mo-MuLV in NIH 3T3 cells (A. Rein and A. Schultz, Virology 136:144-152, 1984). We have now shown that the block to Mo-MuLV infection of MDTF cells occurs at the level of the envelope-receptor interaction. We have cloned the ecotropic receptor cDNA from MDTF cells (dRec) and compared its sequence with that of the NIH 3T3 cell receptor (mRec). Although the deduced dRec and mRec proteins differ at only four amino acid residues, we demonstrate that these changes account for the resistance of MDTF cells to Mo-MuLV infection. Our findings suggest that retroviruses in the same receptor class can exhibit different host ranges due to single amino acid differences in their cellular receptor.

3T3 Cells↗
PubMed1993

Gibbon ape leukemia virus and the amphotropic murine leukemia virus 4070A exhibit an unusual interference pattern on E36 Chinese hamster cells.

The gibbon ape leukemia virus (GaLV), the amphotropic mouse leukemia virus (A-MLV) 4070A, and the xenotropic mouse leukemia virus (X-MLV) exhibit wide but not identical species host ranges. However, most Chinese hamster cells resist infection by all three viruses. We have now determined that the Chinese hamster cell line E36 differs from other Chinese hamster cell lines in that it is susceptible to infection by wild-type GaLV, A-MLV, and X-MLV. Surprisingly, analysis of the interference pattern of GaLV and A-MLV in E36 cells indicated that GaLV and A-MLV interfere in a nonreciprocal fashion. E36 cells productively infected with GaLV were resistant to superinfection by both GaLV and amphotropically packaged recombinant retroviral vectors. In contrast, E36 cells infected with A-MLV were resistant to superinfection with an amphotropic vector but could still be infected by a GaLV vector. These results imply the existence of a receptor on E36 cells that interacts with both GaLV and A-MLV.

3T3 Cells↗
PubMed1993

Interaction of oestradiol, alpha-melanotrophin and noradrenaline within the ventromedial nucleus in the control of female sexual behaviour.

A possible inter-relationship between oestrogen, alpha-melanotrophin (alpha MSH) and NA in the ventromedial nucleus (VMN) has been studied in ovariectomised-adrenalectomised rats primed with a low dose of oestradiol benzoate (2 or 5 micrograms OB), which induces receptivity in approximately half the rats. Priming with OB increased NA turnover in the VMN (as assessed by the decline in NA concentration 1 h after 250 mg/kg alpha-methyl tyrosine alpha MT) and also enhanced the release of NA from basal medial hypothalamic fragments in vitro. This occurred whether the rats were receptive or non-receptive. Injection of 20 micrograms/rat alpha MSH, 4 h before autopsy in OB-primed rats, reduced NA turnover in the VMN but only in the receptive animals. alpha MSH had no effect on NA content in the VMN, but prevented the decline normally induced by the alpha MT, indicating an inhibition of release. Application of 1 microgram/ml alpha MSH to incubated hypothalamic fragments enhanced release of NA in the tissue obtained from untreated controls and the OB-non-receptive group, but had no effect on the tissue of the OB-receptive animals. Perhaps NA release had already occurred in vivo in the latter group. alpha MSH (100 ng/side/rat) and NA (20, 200 and 2,000 ng/side/rat) were injected into the VMN of ovariectomised-adrenalectomised rats primed with 1 microgram OB. Both agents stimulated lordosis in non-receptive animals with a peak activity at 60 min.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Restorative dental care of the geriatric patient.

Dental care of the elderly generally does not form a major area of study in the undergraduate curriculum, and seldom forms the main attraction at dental conferences. All too often treatment of the elderly is considered to be a backwater of prosthetic dentistry and little attention is paid to the restorative needs of this rapidly growing section of society. The aim of this paper is to identify the dental needs of an ageing population, and to focus the attention of the profession on these needs as well as to explore certain myths and prejudices associated with the treatment of patients in senescence.

Aged↗
PubMed1993

Mullerian inhibiting substance requires its N-terminal domain for maintenance of biological activity, a novel finding within the transforming growth factor-beta superfamily.

Mullerian inhibiting substance (MIS)/anti-Mullerian hormone is a differentiation factor that causes regression of the Mullerian duct in the developing male fetus and an apparent sex reversal of the fetal ovary when inappropriately exposed to it. The purified product is a 140-kilodalton glycoprotein composed of two identical subunits. We show that a C-terminal fragment of MIS, which shares homology with transforming growth factor-beta, causes regression of the Mullerian duct and inhibits the biosynthesis of aromatase in the fetal ovary. However, both activities are enhanced dramatically by addition of the N-terminal portion of MIS. Under conditions where potentiation occurs, the N- and C-terminal domains of MIS reassociate. These results indicate that the N-terminus of MIS, unlike that of the other members of the transforming growth factor-beta family, plays a role in maintaining the biological activity of the C-terminus.

Animals↗
PubMed1993

Hormonal control of ovarian phospholipase A2 activity in rats.

The ovulatory effect of LH is mediated by prostaglandins, the synthesis of which involves prostaglandin synthetase and phospholipase A2 (PLA2). It is known that the activity of the former is regulated by LH in the ovary and the objective of this study was to establish the presence of PLA2 in the rat ovary and to determine whether LH has a role in regulating its activity. We demonstrated the presence of a calcium-dependent PLA2 in rat ovaries and shown that, in adult rats, the activity of this enzyme rises significantly on the evening of pro-oestrus (at the expected time of the preovulatory LH surge) and remains high over the evening hours, declining by the next morning. Furthermore, pregnant mares' serum gonadotrophin (PMSG), which when injected into 28-day-old rats induces an LH surge 54 h later, caused at the same time, a rise in ovarian PLA2 activity. Administration of 5 micrograms oestradiol benzoate S.C. to 25-day-old rats stimulated ovarian PLA2 activity 54 h later, at the time of the oestrogen-induced LH surge. Human chorionic gonadotrophin (hCG) had no effect on circulating concentrations of LH, FSH or oestradiol but stimulated ovarian PLA2 activity 2 h later. Gonadotrophin-releasing hormone (GnRH) given to 25-day-old rats (either untreated or primed with 0.5 mg oestradiol benzoate) had no effect on ovarian PLA2 activity but the resulting LH surge may have been too transient.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗