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M R Wilson

Publications and source records attributed to M R Wilson.

At least 73 records · Page 4Linked to original sources
PubMed1998

Evaluation of the contraceptive potential of recombinant human ZP3 and human ZP3 peptides in a primate model: their safety and efficacy.

PROBLEM: The unique recognition events that result in the avid binding of mammalian spermatozoa to the surface of the zona pellucida (ZP) are being exploited in the development of contraceptive vaccines. In this study, the safety and efficacy of a vaccination strategy based on the induction of active immunity against purified, glycosylated, recombinant human ZP3 (rhZP3) has been evaluated in a primate model, Callithrix jacchus. METHOD OF STUDY: Long-term infertility was established after immunization with rhZP3 and the resulting immune sera reacted with rhZP3 on an enzyme-linked immunosorbent assay (ELISA) and immunolocalized exclusively to the outer surface of native ZP on marmoset ovarian sections. However, this contraceptive effect was inevitably associated with the eventual appearance of an ovarian pathology characterized by a depletion of primordial follicles. In an attempt to circumvent this side effect, human ZP3 (hZP3) was epitope mapped and four continuous, immunodominant B-cell epitopes (hZP3(45-64), hZP3(93-110), hZP3(172-190) and hZP3(341-360) were evaluated for contraceptive efficacy in vivo. Using peptide-tetanus toxoid (TT) conjugates to enhance immunogenicity, antipeptide antibodies were raised against these immunogens, which also cross-reacted with rhZP3 on ELISA. In addition, antibodies against hZP3(45-64) and hZP3(172-190) recognized native ZP on marmoset ovarian sections when a microwave technique was used to enhance epitope presentation. RESULTS: No ovarian pathology was observed after the long-term administration of these peptide immunogens, and fertility was suppressed when compared with TT controls but could not be correlated to the antibody titer. CONCLUSION: Clearly, further research is required to identify optimal B-cell epitopes that will reliably induce infertility, free from any ovarian pathology.

Amino Acid Sequence↗
PubMed1998

Apoptotic signal transduction: emerging pathways.

Apoptosis is a counterbalance to mechanisms of cell proliferation and is critically important in regulation of the immune system, development, and normal tissue homeostasis. Mammalian signal transduction pathways affecting apoptosis are more complex than their counterparts in the nematode Caenorhabditis elegans, a valuable model system that has provided powerful initial insights into key molecules regulating apoptosis. Despite this complexity, substantial progress has been made in recent years towards defining the nature and detail of signalling pathways bringing about apoptosis in mammalian cells. In particular, the identity and precise substrate specificities of a large family of caspase enzymes, implicated as critical components of the apoptotic machinery, have been defined. In addition, the mechanism by which the cell surface Fas receptor mediates induction of apoptosis, via activation of caspases, has recently been elucidated. A prominent role for mitochondria in cell death pathways has also recently emerged, a clear theme being that mitochondria can trigger degradative events by the release of apoptogenic proteins (e.g., cytochrome c) from the intermembrane space to the cytosol. This review focuses on recent progress in these areas and discusses integration of this knowledge in our overall understanding of the processes that control apoptosis.

Animals↗
PubMed1998

Isolation, amplification, and sequencing of human mitochondrial DNA obtained from human crab louse, Pthirus pubis (L.), blood meals.

The ability to identify individual human hosts based on analyses of blood recovered from the digestive tract of hematophagous arthropods has been a long-term pursuit in both medical and forensic entomology. Blood meal individualization techniques can bring important advancements to studies of vector-borne disease epidemiology. Forensically, these analyses may aid in assailant identification in violent crime cases where blood-feeding insects or their excreta are recovered from victims or at crime scenes. Successful isolation, amplification, and sequencing of human mitochondrial DNA obtained from adult human crab lice fed on human volunteers are reported. Adult lice were removed from recruited volunteers frequenting inner city health clinics. Live lice were killed by freezing and subsequently air dried at ambient temperature. A saliva sample was obtained from each volunteer and served as a DNA reference sample. Volunteers were afforded free, approved pediculosis treatment. Individual lice were subsequently processed using procedures developed for the extraction of mitochondrial DNA from human hair, teeth, and bone. The resulting DNA was amplified by the polymerase chain reaction and sequenced. Our results point to valuable avenues for future entomological research.

Animals↗
PubMed1998

Spatial and temporal expression of angiogenic molecules during tumor growth and progression.

The growth and metastasis of cancer directly correlates with tumor angiogenesis. A better understanding of the expression of regulatory factors controlling angiogenesis is important in exploiting this process therapeutically. Our present study demonstrates that small tumors (3-4 mm in diameter) express more basic fibroblast growth factor (bFGF) and interleukin 8 (IL-8) than large tumors (> 10 mm in diameter), whereas more vascular endothelial growth factor (VEGF) is expressed in large tumors. Immunostaining showed a heterogeneous distribution of angiogenic factors within the tumor; expression of bFGF and IL-8 was highest on the periphery of a large tumor, where cell division is maximum. VEGF expression was higher in the center of the tumor. In vitro studies demonstrated that sparse cultures of tumor cells expressed higher levels of bFGF and IL-8 than confluent cultures. In contrast, the expression of bFGF and IL-8 was not diminished in tumor cells growing on confluent monolayers of normal cells. VEGF expression was upregulated by cell density irrespective of contact with tumor cells or normal cells. These results demonstrate that the expression of different angiogenic factors in tumor cells can be regulated by their proximity to other tumor cells or host cells.

Animals↗
PubMed1997

Effects of clusterin overexpression on TNFalpha- and TGFbeta-mediated death of L929 cells.

Clusterin is a widely distributed and highly conserved protein for which many functions have been proposed. We used transfected L929 cells to study the effect of clusterin expression on the regulation of cell death signals. We showed that high levels of clusterin expression, about 0.2 pg clusterin secreted per cell per 48 h period, specifically protected L929 cells from TNFalpha-mediated cytotoxicity, while low expression (about 4 fg/cell/48 h) had no effect. However, clusterin expression did not provide transfected L929 cells with protection against death mediated by colchicine, staurosporine or azide. High level expression of clusterin in transfected L929 cells also potentiated the cytotoxicity of TGFbeta. It had previously been shown that exposure of L929 cells to TGFbeta provides protection against TNFalpha. We showed that this protective effect is not additive to that of clusterin expression. One interpretation of this data is that it suggests that clusterin and TGFbeta may act via a common mechanism to provide protection against the cytotoxicity of TNFalpha. Our results indicate that an intracellular action of clusterin protein is responsible for protection against TNFalpha cytotoxicity. Exposure to TNFalpha induces an increase in the level of cell-associated clusterin and specifically in the level of a novel clusterin molecule, which when analyzed under reducing conditions by SDS/PAGE and immunoblotting appears as two closely spaced bands at about 36 and 38.5 kDa. When analyzed under the same conditions, the normal form of intracellular clusterin, which is present with or without exposure to TNFalpha, appears as two poorly resolved bands at about 43-45 kDa. Since the novel form of clusterin is also expressed in cells exposed to TGFbeta, colchicine, staurosporine, and azide, it may result from toxin-induced disruption of processes of normal cellular protein production.

Animals↗
PubMed1997

Increasing metastatic potential is associated with induced chromosome 14 translocations in a previously nonmetastatic murine melanoma cell line.

The purpose of these studies was to demonstrate causal effects of abnormalities induced in mouse chromosome 14 on tumorigenicity and metastasis using the K-1735 murine melanoma cell line. Because anomalies in chromosome 14 have previously been associated with increases in metastatic potential, we induced chromosome 14 anomalies in a nonmetastatic K-1735 clone 10 cells initially containing two normal copies of chromosome 14 by treatment with mitomycin C. Clone 10-M1, in which a small population of cells (approximately 4%) contained translocations involving chromosome 14, was isolated and injected into athymic nude mice. Unlike the parental C-10 cells, C-10 M1 cells produced experimental lung metastases. Chromosomal analysis of cell cultures established from both subcutaneous tumors and lung metastases indicated that approximately 35% of the cell population contained chromosome 14 anomalies suggesting that this chromosome may play a role in tumor growth and metastasis.

Animals↗
PubMed1997

Spanish population data on seven loci: D1S80, D17S5, HUMTH01, HUMVWA, ACTBP2, D21S11 and HLA-DQA1.

Blood samples from 120 Spanish Caucasian individuals were amplified and typed by electrophoresis at six loci, and by reverse dot-blot hybridization at one locus. Results demonstrate the assumption of independence within and between the seven loci analyzed. Therefore, a Spanish population database has been established and statistical analysis shows that a high degree of discrimination can be obtained when all seven (or fewer) loci are used to characterize forensic biological evidence.

DNA↗
PubMed1997

Protection of herpes simplex virus thymidine kinase-transduced cells from ganciclovir-mediated cytotoxicity by bystander cells: the Good Samaritan effect.

Although considerable attention has been directed in the field of gene therapy toward elucidating the mechanism by which a transduced cell could kill a bystander cell, little is known about how bystander cells may affect transduced cells. We hypothesized that bystander cells, particularly if they were capable of gap junctional communication, could protect cells transduced with the herpes simplex virus thymidine kinase (HSV-TK) from ganciclovir (GCV)-induced cytotoxicity. To test this hypothesis, we used a rat hepatocyte cell line (WB) that can carry out efficient gap junctional communication, a WB clone transduced with HSV-TK (WB-TK), and a communication-incompetent subclone of WB cells (aB1). We cocultured WB-TK cells with either WB or aB1 cells, treated them with GCV, and then plated the cells into selective media that permitted us to quantify independently the surviving fraction of WB-TK cells or bystander cells. We found that WB bystander cells conferred up to a 1000-fold protection on WB-TK cells treated with GCV. aB1 cells conferred detectable, but significantly less, protection. These findings demonstrate that herpes simplex virus thymidine kinase-transduced cells can be significantly protected by bystander cells, particularly those that can carry out gap junctional communication. Whether this "Good Samaritan" effect improves the overall efficacy of gene therapy, by prolonging the survival of the source of toxic metabolites, or decreases effectiveness by increasing the survival of transduced cells will need to be determined in vivo.

Animals↗
PubMed1997

Initial clinical experience with the Ahmed Glaucoma Valve implant in pediatric patients.

OBJECTIVE: To evaluate the Ahmed Glaucoma Valve implant, an aqueous shunting device with a unidirectional valve mechanism, in patients younger than 18 years. DESIGN: Prospective case series. SETTING: Tertiary care hospital. PATIENTS: Twenty-one consecutive patients younger than 18 years. The median age of patients was 4.8 years (range, 0.23-17.9 years) INTERVENTION: Placement of an Ahmed Glaucoma Valve implant between April 1992 and April 1994. MAIN OUTCOME MEASURE: Time after surgery without failure. Success was defined as an average intraocular pressure less than 22 mm Hg for the last 2 follow-ups in eyes with preoperative intraocular pressure greater than 22 mm Hg, or an intraocular pressure that was lowered by at least 20% from preoperative values in eyes with preoperative intraocular pressure less than 22 mm Hg, and no additional glaucoma surgeries or visually devastating complications. RESULTS: Cumulative probabilities of success at 12 and 24 months were 77.9% +/- 8.8% and 60.6% +/- 13.7%, respectively. One eye had a flat anterior chamber and suprachoroidal hemorrhage on the first postoperative day. No other eyes had flat or shallow anterior chambers that required reformation. In 3 eyes the implant extruded from underneath the conjunctiva and was removed. In 2 other eyes the average intraocular pressure for the last 2 follow-ups was greater than 22 mm Hg. In 1 eye with an intraocular pressure less than 22 mm Hg preoperatively, the intraocular pressure was not reduced by at least 20%, although the number of antiglaucoma medications was reduced. CONCLUSION: The 12-and 24-month success rates of the Ahmed Glaucoma Valve implant are similar to those of other implants when used in a pediatric population.

Adolescent↗
PubMed1997

Influence of glaucomatous visual field loss on health-related quality of life.

We examined the influence of glaucomatous visual field defects on vision-targeted and generic health-related quality of life. Vision-targeted and generic health status were assessed across 5 glaucoma treatment categories and a normal reference group from 5 tertiary care ophthalmology practices during regularly scheduled eye care visits. The sample consisted of 147 patients who were members of specific glaucoma treatment categories and 44 reference group patients. For patients with glaucoma, eligibility included a diagnosis of glaucoma at least 1 year prior to enrollment and no evidence of other eye disease. Participants completed 2 vision-targeted surveys, the National Eye Institute Visual Functioning Questionnaire and the VF-14, and a generic health-related quality of life measure, the Medical Outcomes Study 36-Item Short Form. Data from automated perimetry (Humphrey Field Analyzer 24-2, Humphrey Instruments, San Leandro, Calif) were used to generate Advanced Glaucoma Intervention Study scores for all participants. The Medical Outcomes Study 36-Item Short Form scores from glaucoma and reference group participants collected on a random half of the sample were similar. However, comparisons of the vision-targeted surveys demonstrated significant mean differences on 7 of 11 National Eye Institute Visual Functioning Questionnaire scales, and a trend toward significant differences for the VF-14 (P < .07 by linear regression). Greater visual field defects in the better eye were significantly associated with poorer National Eye Institute Visual Functioning Questionnaire scores (P < .05), as well as with worse VF-14 scores. These findings were most dramatic for patients with the most severe field loss in the better eye. Vision-targeted questionnaires were more sensitive than a generic health-related quality of life measure to differences between glaucoma and normal reference participants. Our findings indicate that self-reports of vision-targeted health-related quality of life are sensitive to visual field loss and may be useful in tandem with the clinical examination to fully understand outcomes of treatment for glaucoma.

Adult↗
PubMed1997

Range of antinuclear antibodies in "healthy" individuals.

OBJECTIVE: To determine the range of antinuclear antibodies (ANA) in "healthy" individuals compared with that in patients with systemic lupus erythematosus (SLE), systemic sclerosis (SSc; scleroderma), Sjögren's syndrome (SS), rheumatoid arthritis (RA), or soft tissue rheumatism (STR). METHODS: Fifteen international laboratories experienced in performing tests for ANA by indirect immunofluorescence participated in analyzing coded sera from healthy individuals and from patients in the 5 different disease groups described above. Except for the stipulation that HEp-2 cells should be used as substrate, each laboratory used its own in-house methodology so that the data might be expected to reflect the output of a cross-section of worldwide ANA reference laboratories. The sera were analyzed at 4 dilutions: 1:40, 1:80, 1:160, and 1:320. RESULTS: In healthy individuals, the frequency of ANA did not differ significantly across the 4 age subgroups spanning 20-60 years of age. This putatively normal population was ANA positive in 31.7% of individuals at 1:40 serum dilution, 13.3% at 1:80, 5.0% at 1:160, and 3.3% at 1:320. In comparison with the findings among the disease groups, a low cutoff point at 1:40 serum dilution (high sensitivity, low specificity) could have diagnostic value, since it would classify virtually all patients with SLE, SSc, or SS as ANA positive. Conversely, a high positive cutoff at 1:160 serum dilution (high specificity, low sensitivity) would be useful to confirm the presence of disease in only a portion of cases, but would be likely to exclude 95% of normal individuals. CONCLUSION: It is recommended that laboratories performing immunofluorescent ANA tests should report results at both the 1:40 and 1:160 dilutions, and should supply information on the percentage of normal individuals who are positive at these dilutions. A low-titer ANA is not necessarily insignificant and might depend on at least 4 specific factors. ANA assays can be a useful discriminant in recognizing certain disease conditions, but can create misunderstanding when the limitations are not fully appreciated.

Adult↗
PubMed1997

A family exhibiting heteroplasmy in the human mitochondrial DNA control region reveals both somatic mosaicism and pronounced segregation of mitotypes.

A family exhibiting heteroplasmy at position 16355 in hypervariable region I of the human mtDNA control region has been identified. This family consists of a mother, daughter, and son. DNA samples extracted from blood stains, buccal swabs, and hairs from these individuals were amplified by PCR and sequenced utilizing fluoresence-labeled dye terminator chemistry in an automated DNA sequencer. In both the daughter and mother, heteroplasmy was observed in DNA extracted from blood stains, buccal swabs, and hairs. In the blood stains, the proportion of cytosine was greater than thymine in both individuals. Buccal swab extracts showed a more balanced contribution from the two nucleotides. Telogenic hair root and hair shaft samples exhibited a wide range of nucleotide contributions at this position, from predominately cytosine in some samples to predominately thymine in others. The apparent stochastic segregation of mitotypes in hair samples is discussed from a forensic viewpoint, and the mechanism of mtDNA heteroplasmy is considered.

DNA, Mitochondrial↗
PubMed1997

Clinical experience with the Ahmed Glaucoma Valve implant in eyes with prior or concurrent penetrating keratoplasties.

PURPOSE: To evaluate the Ahmed Glaucoma Valve implant, an aqueous shunting device with a unidirectional valve mechanism, in eyes with concurrent or prior penetrating keratoplasties. METHODS: Thirty-one eyes of 31 consecutive patients had placement of an Ahmed Glaucoma Valve implant. Median patient age was 65.1 years (range, 17.2 to 103.4 years). The main outcome measure was time after surgery without failure. Success was defined as no additional glaucoma surgeries or devastating visual complications, no new corneal graft failure, an intraocular pressure greater than or equal to 5 mm Hg on the last two follow-up examinations, and reduction in intraocular pressure. For eyes with preoperative intraocular pressure greater than 22 mm Hg, an average intraocular pressure of less than 22 mm Hg on the last two follow-up examinations was required. For eyes with preoperative intraocular pressure of less than 22 mm Hg, an intraocular pressure lowered by at least 20% from preoperative values was required. RESULTS: Cumulative probabilities of success at 12 and 20 months (mean +/- SD) were 75.4% +/- 8.2% and 51.5% +/- 11.4%, respectively. Eleven of 31 eyes were failures. The risk of failure in eyes with prior infectious keratitis or keratouveitis was estimated to be 5.8 times greater than that associated with eyes that underwent penetrating keratoplasties for other reasons (P = .009). CONCLUSIONS: Twelve- and 20-month success rates of the implant in eyes with prior or concurrent penetrating keratoplasties were comparable to those of other drainage devices. Eyes with prior infectious keratitis or keratouveitis were at increased risk of failure.

Adolescent↗
PubMed1997

A high observed substitution rate in the human mitochondrial DNA control region.

The rate and pattern of sequence substitutions in the mitochondrial DNA (mtDNA) control region (CR) is of central importance to studies of human evolution and to forensic identity testing. Here, we report a direct measurement of the intergenerational substitution rate in the human CR. We compared DNA sequences of two CR hypervariable segments from close maternal relatives, from 134 independent mtDNA lineages spanning 327 generational events. Ten substitutions were observed, resulting in an empirical rate of 1/33 generations, or 2.5/site/Myr. This is roughly twenty-fold higher than estimates derived from phylogenetic analyses. This disparity cannot be accounted for simply by substitutions at mutational hot spots, suggesting additional factors that produce the discrepancy between very near-term and long-term apparent rates of sequence divergence. The data also indicate that extremely rapid segregation of CR sequence variants between generations is common in humans, with a very small mtDNA bottleneck. These results have implications for forensic applications and studies of human evolution.

Animals↗
PubMed1997

The effect of maternal protein deficiency during pregnancy and lactation on glucose tolerance and pancreatic islet function in adult rat offspring.

To test the hypothesis that poor foetal-neonatal nutrition predisposes adult animals to impaired glucose tolerance or diabetes, pregnant and lactating rats were fed a low (5%) protein diet and glucose tolerance and pancreatic islet function then assessed in the adult offspring. To expose any underlying defects the offspring were allowed access to a sucrose supplement (35%) or fed a high fat diet. Offspring born to low protein-fed females had significantly lower body weights than controls. In islets from previously malnourished rats, insulin release in batch incubations or perifusion was not significantly different to controls. In islets from previously malnourished animals fed sucrose, glucose-stimulated insulin release was reduced in perifusion by 66% (P < 0.01) and batch incubations by 26-52% (6-16 mmol/l glucose, P < 0.01). Similarly, impaired secretory responses were found in islets from previously malnourished animals fed a high fat diet. These did not result from a reduced pool of releasable insulin, as arginine-stimulated secretion was not impaired. Rats previously malnourished showed a normal glucose tolerance. Glucose tolerance was impaired, however, in previously malnourished rats fed sucrose (area under the glucose tolerance test curve was increased by 42%, P < 0.05) but despite the reduced islet secretory responses was not significantly different to sucrose-fed controls (area increased by 54%, P < 0.05). Glucose tolerance was impaired in previously malnourished animals fed high fat diet (area increased by 48%, P < 0.05) more so than in high fat fed-controls (28% increase, NS). These data support the hypothesis that poor foetal-neonatal nutrition leads to impaired pancreatic beta-cell function which persists into adult life. Alone this is not sufficient to produce diabetes, but an inability to respond to a highly palatable fat diet may tip the balance towards impaired glucose tolerance.

Animals↗
PubMed1997

Melorheostosis.

Melorheostosis is a rare, benign connective-tissue disorder characterized by a cortical thickening of bone with irregular dense hyperostosis that appears to flow along the cortex. We review the literature to date, etiology, clinical aspects, roentgenographic features, histology, and treatment of melorheostosis.

Bone Diseases, Metabolic↗