Biomedical subjects

M J Anderson

Publications and source records attributed to M J Anderson.

At least 19 recordsLinked to original sources

PubMed1995

Erratic deposition of agrin during the formation of Xenopus neuromuscular junctions in culture.

In order to disclose the mechanism that regulate synapse development we compared the distributions of agrin, acetylcholine receptors (AChR) and a basal lamina heparan sulfate proteoglycan (HSPG) in sections and cultures prepared from Xenopus laevis and Ambystoma mexicanum embryos. While agrin, AChR and HSPG may accumulate almost synchronously at synapses in vivo, agrin deposition usually lagged well behind the other synaptic markers during development in culture, and was not detectable at many differentiated junctions. Agrin deposition at nerve-muscle contacts in culture also appeared to require the presence of other synaptic components. A similarly variable deposition occurred on noninnervated myocytes, where agrin again collected near sites of HSPG and AChR accumulation on some cells. Profuse agrin accretion occurred consistently, however, within the extracellular matrices of surrounding epithelial cells derived from both myotomes and neural tubes. In cocultures of Ambystoma neurons and Xenopus myocytes Ambystoma agrin collected at some chimeric neuromuscular junctions, but also accumulated on noninnervated myocytes and in the extracellular matrices of salamander neuroendothelial cells. Based upon these observations we conclude that (a) focal agrin deposition is not required for synaptic differentiation on Xenopus myocytes and (b) agrin may be one of several muscle basal lamina components that stem mainly from the secreted products of nearby epithelial cells.

Agrin↗

PubMed1995

Molecular typing of Aspergillus species.

In the last four years, several molecular typing methods have been applied to various Aspergillus species, primarily A. fumigatus and A. niger. Many 'clones' or DNA types exist in the environment and in patients. The most discriminatory methods appear to be restriction fragment length polymorphism analysis with or without specific probes and random amplification of polymorphic DNA analysis. Insufficient work on interlaboratory reproducibility has been done. Most patients are infected by single DNA types, although occasionally multiple isolates are detected. Little work has been done with regard to detailed hospital epidemiology using molecular typing methods.

Aspergillus↗

PubMed1995

The relationship between carbon dioxide sensitivity and sprint or endurance performance in young swimmers.

There has been some evidence that extremes of CO2 sensitivity can indicate an individual's potential for sprint or endurance athletic performance. This study examined the responses to CO2 rebreathing, and previously validated sprint and endurance tests by subjects who were involved in regular, but not intensive, swimming training. The aim was to determine whether subjects with low CO2 sensitivity might perform better on endurance tests, whilst those subjects with high CO2 sensitivity would be more successful in the sprint tests. Initially, 168 young (mean mean(s.d.) age = 12.4(2.1) years) swimmers were measured using a modified version of the Read CO2 rebreathing technique. From this sample, 17 high (mean means(s.d.) = 2.24(0.39) 1 min-1 mmHg-1) CO2 responders were matched by gender, age, height, weight and FVC with 17 low (mean means(s.d.) = 0.57(0.19) 1 min-1 mmHg-1) responders. Each of these 17 pairs underwent two sprint tests (10 s Tri-level alactic power, 50 m run) and two endurance tests (PWC170, 1.6 km run) in order to determine whether any differences existed between the two groups and the sprint and endurance parameters. The subjects remained unaware of their results of the rebreathing test throughout testing. A dependent t test was then used to compare the results collected from each group. The low CO2 responders recorded a significantly faster 1.6 km run time, but were not superior on the PWC170 ergometer test, than the high CO2 responders. The high CO2 responding group recorded significantly higher results on the 10 s alactic power test, but not the 50 m sprint run, than the low responders. Hence, further study is required to ascertain underlying causes as to why significant differences occurred with CO2 rebreathing, the 1.6 km run and the 10 s alactic power test, but not with the PWC170 and the 50m sprint run.

Adolescent↗

PubMed1995

The relationship of hypercapnic ventilatory responses to age, gender and athleticism.

When a gas mixture containing carbon dioxide (CO2) is inhaled by an individual, alveolar ventilation is increased. This ventilatory response to CO2 highlights the relationship between the increase in exercise ventilation and the increase in alveolar CO2 pressure (paCO2). This response is mediated centrally by brainstem chemoreceptors in the medulla and, to a lesser extent, peripherally by the carotid and aortic bodies. However, the response of increased breathing to rising paCO2 varies markedly among individuals. The responses to CO2 rebreathing have been investigated by a variety of research groups for different reasons. The range of responses by children and adults including: gender differences; responses by athletes; the relationship between age and body size; and whether it is an acquired or inherited response have been studied. The following is a summary of these different aspects of CO2 rebreathing has been complied to assist researchers studying any single or multiple facets of the area.

Adolescent↗

PubMed1995

Total hip replacement for developmental dysplasia of the hip.

Total hip arthroplasty relieves pain and improves function for many patients with endstage arthritis secondary to developmental dysplasia of the hip. Acetabular dysplasia, however, presents a special problem for total hip reconstruction in these patients. Structural bone grafting with femoral head autografting to the dysplastic acetabulum and cementing acetabular components into the graft provides satisfactory short-term results, but longer-term followup data show high acetabular component failure rates. At a mean followup period of 7 years, 20% of components were loose, and at a mean followup period of 12 years, 46% were loose. Noncemented porous-coated hemispherical acetabular components have considerably expanded the success of total hip replacement without the need for structural bone grafting in such patients. At a mean followup period of almost 7 years, none of the acetabular components were reported to be loose in 1 series. For most patients, the acetabular dysplasia can be managed by techniques such as reaming deeper, using small-diameter porous-coated acetabular components, using screws to provide rigid initial stability for the components, increasing the height of the prosthetic hip center, and covering small portions of the components with bone graft chips if necessary.

Hip Dislocation↗

PubMed1994

Chromosome 13 transfer provides evidence for regulation of RB1 protein expression.

The human retinoblastoma susceptibility gene (RB1) located on chromosome 13 has been shown to function as a growth/tumor suppressor gene in a large number of human cancers. Although constitutive expression has been observed in most cultured cells and normal tissues, overexpression of RB1 protein has not been well documented. Perhaps regulating the level of normal RB1 protein expression is one of several ways of controlling its function. To test this hypothesis, we transferred normal copies of chromosome 13 via microcell fusion into the human fibrosarcoma cell line HT1080. Microcell hybrids were generated that contained one, two, or three extra copies of the transferred fibroblast chromosome 13. Compared to the parental cell line, the hybrids were completely unaltered with respect to several properties in vitro and in vivo, including morphology, growth rate, and tumor formation. Northern blot analysis revealed a stepwise increase in RB1 mRNA expression which increased in proportion to the number of alleles present in each cell line. Although RB1 protein exhibited correct nuclear localization and was phosphorylated in a normal cell cycle-dependent manner in the hybrids, the increased level of protein expression in each hybrid was nearly identical and did not increase beyond a threshold amount, although mRNA expression continued to increase. These results demonstrate that HT 1080 cells can tolerate an increase level of RB1 protein, but that expression beyond a certain level may be down-regulated. These transfer studies provide evidence for regulation of RB1 protein expression and may suggest an alternative form of monitoring and controlling normal RB1 functioning.

Animals↗

PubMed1994

Evidence that wild-type TP53, and not genes on either chromosome 1 or 11, controls the tumorigenic phenotype of the human fibrosarcoma HT1080.

The specific transfer of normal chromosomes via microcell fusion has been instrumental in identifying putative tumor suppressor gene loci in a variety of human cancers. Using this same technique it has been proposed that the tumorigenicity of the human fibrosarcoma cell line HT1080 is controlled by functionally distinct tumor suppressor genes on human chromosomes I and II. To address these results and perhaps further localize the suppressive effect to particular regions on these two chromosomes, we transferred into HT1080 seven different fibroblast-derived human chromosomes containing either intact or discrete portions of chromosome I or II. Interestingly, we found no evidence of genes on these chromosomes that could alter the growth of HT1080 either in vitro or in vivo. Based on these results we were left with the possibility that a gene, or genes, residing on an entirely different chromosome(s) was involved in the tumorigenesis of HT1080. Since TP53 mutation has been documented in a variety of human tumor types, and we found both copies of TP53 to be mutated in HT1080, we were prompted to examine its role by both cDNA transfection and chromosome transfer. Although by cDNA transfection we found that expression of exogenous wild-type TP53 was incompatible with continued proliferation of HT1080 cells in vitro, chromosome 17 transfer studies revealed that a more physiologic expression of exogenous wild-type TP53 could be tolerated in vitro while being completely incompatible with growth in vivo. These studies demonstrate a differential effect of TP53 growth inhibition and clearly show that TP53 tumor suppressing function can be independent from its potent growth suppressing effect in vitro.

Animals↗

PubMed1994

Effect of dexamethasone and oxygen exposure on neonatal rat lung retinoic acid receptor proteins.

Retinol deficiency in animal models results in histopathologic airway changes that appear similar to those found in human premature infants with bronchopulmonary dysplasia (BPD). Dexamethasone (DEX), a steroid now often used in the treatment of BPD, might potentially affect lung vitamin A homeostasis since it alters serum and liver retinoid stores in certain models. Our objective was to determine the effect of DEX on neonatal rat lung retinoid status and the binding of retinoic acid (RA) to cytosolic and nuclear receptor proteins. We examined this effect both in room air and when the animals breathed 95% oxygen (O2). Twenty-four 1-day-old rat pups received either 1 microgram/g DEX subcutaneously, an equal volume of normal saline (NS) subcutaneously at 0 (start experiment time), 24, and 48 hours, or no injection at all, and were sacrificed at 72 hours. Twelve rats in each treatment group were housed in room air and 12 in each group were exposed to > 95% O2 for the 3 day period. Lung and liver were analyzed for retinyl palmitate (RP). Nuclear retinoic acid receptor (RAR) and cellular retinoic acid binding protein (CRABP) were measured by specific binding assays. DEX decreased liver RP by 33-55% and rat pup lung RP by over 60%; it also decreased lung RAR binding (mean dpm/microgram protein +/- SEM) in both room air and oxygen groups: Air (11.2 +/- 1.0) vs. Air/DEX (4.6 +/- 1.3, n = 6; P < 0.01), and O2 (18.2 +/- 0.6) vs. O2/DEX (3.2 +/- 0.6, n = 6; P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Neuronal differentiation in vitro from precursor cells of regenerating spinal cord of the adult teleost Apteronotus albifrons.

This study documents neuronal differentiation in vitro from undifferentiated precursor cells of caudalmost regenerating spinal cord of the teleost Apteronotus albifrons. At 11 days in vitro, cells from the caudalmost tip of the regenerating cord are flat and polygonal in shape, lack neuronal processes and do not stain with antibody against neuron-specific filaments. At 15 days in vitro, some of the caudalmost cells have developed short, neurite-like processes; at 18 days in vitro, some cells react positively with antibody against neuron-specific filaments. At 26 days in vitro, many of the caudalmost cells have long branching neurites and react positively with anti-neurofilament antibody. Addition of insulin-like growth factor-I to the medium accelerates the process of neuronal differentiation from the caudalmost precursor cells in vitro. The source of these precursor cells is ultimately cells of the ependymal layer of adult spinal cord. Further investigation of the factors that control production and differentiation of these cells will be important in defining the developmental potential possible for vertebrate spinal cord cells and may aid in creating an optimal environment for regeneration of axons within mammalian spinal cord.

Animals↗

PubMed1994

Constrained acetabular components.

Between May 1987 and October 1990, 21 constrained acetabular components were used in revision total hip arthroplasty at the University of California, San Francisco. In 18 patients, the device was placed for a chronically dislocating total hip arthroplasty. In the remaining three, intraoperative instability during revision total hip arthroplasty necessitated its use. At the minimum 2-year follow-up evaluation (average, 31 months; range, 24-64 months), 15 patients (71%) experienced no further dislocations or subluxations. There were eight dislocations in the remaining six patients (29%). The average Harris hip score at the follow-up evaluation was 76 points (range, 32-100 points). For those patients who redislocated (n = 6), an increased acetabular abduction angle of the metallic acetabular cup, averaging 70 degrees, was the only predictive factor of failure of the constrained cup (P < .05). No radiographic or clinical evidence of loosening in the 19 porous ingrowth acetabular component was observed. This device will relieve the complication of severe hip instability in the majority of patients, but is not universally successful.

Acetabulum↗

PubMed1994

Characterisation of the novel gene G11 lying adjacent to the complement C4A gene in the human major histocompatibility complex.

Twelve transcriptional units have now been located in a 160 kb segment of DNA that includes the genes encoding members of the serum complement system C2, Factor B (Bf) and C4 within the class III region of the human major histocompatibility complex (MHC). The common arrangement of these genes is tel-C2-Bf-RD-G11-C4A-[P450c21A-YA-XA]-C4B-[P450c21B-YB ]-+ ++TNX-cen. Characterisation of cDNA and genomic clones corresponding to the novel gene G11 has revealed that the gene spans approximately 9.1 kb of DNA and is split into 7 exons. The 5' end of the gene is associated with a CpG-island while the 3' end of the gene lies 611 bp from the transcriptional start site of the C4A gene. The approximately 1.4 kb G11 mRNA, which is expressed in a number of different cell types including monocytes, hepatocytes, epithelial cells, T and B lymphocytes, encodes protein products of 254 or 258 amino acids due to differential use of two splice sites lying 12 bp apart at the end of exon 3. These polypeptides share homology with a limited number of proteins including human cytochrome P450XIB1 and the tyrosine kinase transforming protein from fujinami virus. Duplication of the C4/P450c21 transcriptional unit occurred by a nonhomologous recombination event. Sequence analysis of a 1.5 kb segment of DNA flanking the C4B gene has revealed that 914 bp of the 3' end of the G11 gene also lies 611 bp from the transcriptional start site of the C4B gene.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

PubMed1994

The influence of sometribove dose and days in lactation on behavior of cows implanted with pelleted sometribove.

Data were from observations of behavior of 99 lactating Holstein cows during implantations with bST. Implantations were at 14-d intervals during 1 yr. Cows on one study received 0, 120, 240, or 360 mg; cows on a second study received 0, 160, or 320 mg of bST per implantation. A checklist of behaviors was completed by the recorder and observer for each cow during implantation. Cows were also given an overall temperament score during each session. Lunging occurred in 49% of the sessions, but bellowing, kicking, and kneeling occurred least. More cows scored in the docile range than in the excited range. Cows habituated to head bobbing, back sagging, lunging, moving back in chute, flinching, head up, and bellowing, but not to shifting feet, kicking, shaking the head, head back, or ears back.

Animals↗

PubMed1993

Human parvovirus infection in homozygous sickle cell disease.

We studied the epidemiology of human parvovirus B19 infection in 308 children with homozygous sickle cell (SS) disease and 239 controls with a normal haemoglobin (AA) genotype followed from birth in a cohort study. Annual serum samples identified the time and frequency of B19 infection, which did not differ between SS and AA children, about 40% of each group developing specific IgG by age 15. B19 infection followed an epidemic pattern similar to that observed for aplastic crises; accounted for all 91 aplastic crises that occurred; and was found in an additional 23 SS patients, of whom 10 showed mild haematological changes and 13 no changes. The magnitude or duration of IgG response did not differ between these groups. No patient had 2 attacks of aplasia and no patient nor control had 2 attacks of B19 infection. Following B19 infection, serial specific IgG concentrations remained high after 5 years in only 45% of SS patients, although the rarity of recurrent aplasia suggests lifelong immunity. B19 infection accounts for most if not all aplastic crises in SS disease, but at least 20% of infections do not result in aplasia. An effective vaccine against B19 might make an important contribution to the management of sickle cell disease.

Adolescent↗

PubMed1993

Covalent binding properties of the human complement protein C4 and hydrolysis rate of the internal thioester upon activation.

The complement proteins C3 and C4 have an internal thioester. Upon activation on the surface of a target cell, the thioester becomes exposed and reactive to surface-bound amino and hydroxyl groups, thus allowing covalent deposition of C3 and C4 on these targets. The two human C4 isotypes, C4A and C4B, which differ by only four amino acids, have different binding specificities. C4A binds more efficiently than C4B to amino groups, and C4B is more effective than C4A in binding to hydroxyl groups. By site-directed mutagenesis, the four residues in a cDNA clone of C4B were modified. The variants were expressed and their binding properties studied. Variants with a histidine residue at position 1106 showed C4B-like binding properties, and those with aspartic acid, alanine, or asparagine at the same position were C4A-like. These results suggest that the histidine is important in catalyzing the reaction of the thioester with water and other hydroxyl group-containing compounds. When substituted with other amino acids, this reaction is not catalyzed and the thioester becomes apparently more reactive with amino groups. This interpretation also predicts that the stability of the thioester in C4A and C4B, upon activation, will be different. We measured the time course of activation and binding of glycine to C4A and C4B. The lag in the binding curve behind the activation curve for C4A is significantly greater than that for C4B. The hydrolysis rates (k0) of the thioester in the activated proteins were estimated to be 0.068 s-1 (t1/2 of 10.3 s) for C4A and 1.08 s-1 (t1/2 of 0.64 s) for C4B. These results indicate that the difference in hydrolysis rate of the thioester accounts, at least in part, for the difference in the binding properties of C4A and C4B.

Amino Acid Sequence↗

PubMed1993

Differences in growth of neurons from normal and regenerated teleost spinal cord in vitro.

Explants and dissociated cells from normal adult spinal cord and regenerating cord of the teleost Apteronotus albifrons were grown in vitro for periods of 8 to 12 wk. During this time the neurons showed extensive neurite outgrowth. Neurite outgrowth from tissue explants and dissociated cells of regenerated spinal cord starts sooner and is more profuse than that from normal (unregenerated) cord. Neurite outgrowth is maximized by using adhesive substrata and a high density of explants or dissociated cells. Inasmuch as Apteronotus does regenerate its spinal cord naturally after injury, whereas mammals do not, this culture system will be useful to study factors that control (permit) regeneration of spinal neurons in this adult vertebrate.

Animals↗

PubMed1993

Comparison of the cytology brush with the Dacron swab for detecting Chlamydia trachomatis by enzyme immunoassay in female university students.

Previous studies comparing the cytology brush (Cytobrush) with swabs for the detection of Chlamydia trachomatis from the uterine cervix have yielded divided results. The authors carried out a retrospective analysis at the University of South Florida Student Health Service to evaluate the effect of a change in specimen collection technique on C trachomatis prevalence estimates in female university students. Samples were collected from 1,003 women in 1989, using a small Dacron swab, and 1,229 women in 1990, using a brush; the samples were tested by means of the Pathfinder Enzyme Immunoassay (EIA) detection kit. Positive test results were 5.88% for 1989 and 11.66% for 1990. Actual C trachomatis prevalence for 1989 and 1990 was stable, as determined by three control groups. The results suggest that the cytology brush is more effective than the Dacron swab for the detection of C trachomatis in the uterine cervix when it is used with the Pathfinder EIA test kit.

Adult↗

PubMed1993

Analysis of sequence contexts flanking T.G mismatches leads to predictions about reactivity of the mismatched T to osmium tetroxide.

Osmium tetroxide and hydroxylamine are used to detect mutations in DNA and RNA after hybridization of mutant and wild-type DNA. Mismatched T and C bases, respectively, are modified by these reagents and the DNA strand cleaved at the mismatched bases by subsequent treatment with piperidine. This allows detection and location of the mutation. Although most T.G mismatches have been reported to be reactive to osmium tetroxide, some have been reported to be unreactive. The aim of this study was to collect and analyze the reactive and unreactive T.G mismatches. We have collected sequence contexts of all reactive and unreactive T.G mismatches for analysis. This involves 10 unreactive T.G mismatches (plus one T.C) and 19 reactive T.G mismatches. Sequence effects of bases surrounding these mismatches must influence this reactivity. There must be many types of such sequence effects. We postulate that because of the dominance of 5' G bases near the T of unreactive T.G mismatches and the absence of 5' G bases in reactive T.G mismatches that the stacking of the 5' G on the mismatched T is the reason for this lack of reactivity in the majority of the cases studied here.

Base Sequence↗

PubMed1993

Tumor suppressor genes studied by cell hybridization and chromosome transfer.

Although the cloning of tumor suppressor genes has proved to be an arduous task, often involving several years of labor intensive cloning strategies, a greater understanding of neoplastic progression will be made once the function and role of these genes have been sorted out. To fully appreciate the state at which this field of research currently is, however, one must understand that the road to tumor suppression was paved by both somatic cell hybridization and chromosome transfer studies. Although regarded by many as laboratory artifact, somatic cell hybridization has provided strong circumstantial evidence, if not formal proof, for the existence of tumor suppressor genes. In further reducing the complexity associated with whole genome transfer, single chromosome transfer was subsequently developed as a refinement to this technique so that one could unequivocally correlate suppression with a particular chromosome. We have learned from these studies that single chromosomes harbor the genetic information necessary to reverse the malignant phenotype associated with cancer cells. Furthermore, multiple tumor suppressor loci are now known to exist, with one to several different loci associated with a given tumor type. In this review, we present evidence for tumor suppressor genes and discuss the early studies that led to their discovery.

Animals↗