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Biomedical subjects

A L Watson

Publications and source records attributed to A L Watson.

At least 19 recordsLinked to original sources
PubMed2000

The quality of merC, a module of the mer mosaic.

We examined a region of high variability in the mosaic mercury resistance (mer) operon of natural bacterial isolates from the primate intestinal microbiota. The region between the merP and merA genes of nine mer loci was sequenced and either the merC, the merF, or no gene was present. Two novel merC genes were identified. Overall nucleotide diversity, pi (per 100 sites), of the merC gene was greater (49.63) than adjacent merP (35.82) and merA (32.58) genes. However, the consequences of this variability for the predicted structure of the MerC protein are limited and putative functional elements (metal-binding ligands and transmembrane domains) are strongly conserved. Comparison of codon usage of the merTP, merC, and merA genes suggests that several merC genes are not coeval with their flanking sequences. Although evidence of homologous recombination within the very variable merC genes is not apparent, the flanking regions have higher homologies than merC, and recombination appears to be driving their overall sequence identities higher. The synonymous codon usage bias (EN(C)) values suggest greater variability in expression of the merC gene than in flanking genes in six different bacterial hosts. We propose a model for the evolution of MerC as a host-dependent, adventitious module of the mer operon.

Amino Acid Sequence↗
PubMed2000

Onset of maternal arterial blood flow and placental oxidative stress. A possible factor in human early pregnancy failure.

The aim was to measure changes in the oxygen tension within the human placenta associated with onset of the maternal arterial circulation at the end of the first trimester of pregnancy, and the impact on placental tissues. Using a multiparameter probe we established that the oxygen tension rises steeply from <20 mmHg at 8 weeks of gestation to >50 mmHg at 12 weeks. This rise coincides with morphological changes in the uterine arteries that allow free flow of maternal blood into the placenta, and is associated with increases in the mRNA concentrations and activities of the antioxidant enzymes catalase, glutathione peroxidase, and manganese and copper/zinc superoxide dismutase within placental tissues. Between 8 to 9 weeks there is a sharp peak of expression of the inducible form of heat shock protein 70, formation of nitrotyrosine residues, and derangement of the mitochondrial cristae within the syncytiotrophoblast. We conclude that a burst of oxidative stress occurs in the normal placenta as the maternal circulation is established. We speculate that this may serve a physiological role in stimulating normal placental differentiation, but may also be a factor in the pathogenesis of pre-eclampsia and early pregnancy failure if antioxidant defenses are depleted.

Abortion, Spontaneous↗
PubMed1999

Maternal arterial connections to the placental intervillous space during the first trimester of human pregnancy: the Boyd collection revisited.

OBJECTIVE: We sought to determine morphologically when the maternal uterine circulation to the human placenta is established. STUDY DESIGN: We performed a histologic review of 12 early-pregnancy hysterectomy specimens contained within the Boyd Collection, ranging from 43 to 130 days' gestation. RESULTS: Before the eighth week of pregnancy, maternal arterial connections with the intervillous space are restricted to tortuous networks of intercellular spaces. Only after this period can direct channels be observed. Initially, these are of small caliber, but they become sizable and clearly delineated after 11 to 12 weeks. CONCLUSION: The maternal circulation to the placenta must be extremely sluggish before the eighth week of pregnancy but will be gradually established over the next few weeks. It is unlikely to be substantial until at least 12 weeks. The implication is that development of the human fetoplacental unit during most of the first trimester takes place in a low-oxygen environment.

Arteries↗
PubMed1999

Cd(II)-responsive and constitutive mutants implicate a novel domain in MerR.

Expression of the Tn21 mercury resistance (mer) operon is controlled by a metal-sensing repressor-activator, MerR. When present, MerR always binds to the same position on the DNA (the operator merO), repressing transcription of the structural genes merTPCAD in the absence of Hg(II) and inducing their transcription in the presence of Hg(II). Although it has two potential binding sites, the purified MerR homodimer binds only one Hg(II) ion, employing Cys82 from one monomer and Cys117 and Cys126 from the other. When MerR binds Hg(II), it changes allosterically and also distorts the merO DNA to facilitate transcriptional initiation by sigma70 RNA polymerase. Wild-type MerR is highly specific for Hg(II) and is 100- and 1, 000-fold less responsive to the chemically related group 12 metals, Cd(II) and Zn(II), respectively. We sought merR mutants that respond to Cd(II) and obtained 11 Cd(II)-responsive and 5 constitutive mutants. The Cd(II)-responsive mutants, most of which had only single-residue replacements, were also repression deficient and still Hg(II) responsive but, like the wild type, were completely unresponsive to Zn(II). None of the Cd(II)-responsive mutations occurred in the DNA binding domain or replaced any of the key Cys residues. Five Cd(II)-responsive single mutations lie in the antiparallel coiled-coil domain between Cys82 and Cys117 which constitutes the dimer interface. These mutations identify 10 new positions whose alteration significantly affect MerR's metal responsiveness or its repressor function. They give rise to specific predictions for how MerR distinguishes group 12 metals, and they refine our model of the novel domain structure of MerR. Secondary-structure predictions suggest that certain elements of this model also apply to other MerR family regulators.

Amino Acid Sequence↗
PubMed1999

Academic achievement in adolescent children of alcoholics.

OBJECTIVE: The current study tested whether adolescent children of alcoholics (COAs) showed poorer academic performance than did demographically matched controls, and whether such parent alcoholism effects varied as a function of heterogeneity within the COA sample. In addition, controlling for parent educational attainment, we examined whether relations between parental alcohol dependence and academic performance could be accounted for by COAs' lower levels of task orientation, heightened levels of environmental stress, lowered levels of family organization and less parental involvement in their school activities. METHOD: A sample of 221 adolescent COAs and 196 demographically matched controls (53% boys, mean [+/-SD] age = 12.7+/-1.46 years), and their parents, were included in the current study. Adolescents were selected from a larger 3-year longitudinal study in which participants were interviewed three times at annual intervals. Those who were interviewed at Time 3 and who had academic achievement data were included in the current analyses. Demographic information and diagnoses of parental alcoholism were collected at Time 1, and data on potential mediators were collected at Time 3. Academic achievement data were collected at Time 3 from school records. RESULTS: Multiple regression analyses indicated that COAs received lower school grades than did their non-COA peers (mean = 2.19+/-1.08 vs 2.54+/-1.01, respectively). COAs with two alcoholic parents (mean = 1.80+/-1.17) and COAs with at least one parent diagnosed alcohol dependent (mean = 2.01+/-1.01) showed particularly low grades. Parental alcohol dependence was also associated with lower math achievement scores (mean = 48.52+/-24.68 vs 62.47+/-26.71). Evidence indicated that adolescents' task orientation mediated the relation between parental alcohol dependence and adolescent grades (indirect effect, t = -2.93, 289 df, p < .01), and between parental alcohol dependence and adolescent math achievement (indirect effect, t = -1.99, 194 df, p < .01). Adolescents' life stress did not mediate the relations of interest after controlling for task orientation. CONCLUSIONS: The current study confirmed that COAs, particularly those whose parents are alcohol dependent as opposed to having a diagnosis of alcohol abuse, achieve relatively lower academic outcomes in comparison to non-COA peers. Adolescent task orientation partially mediated the relations between parent alcohol dependence and academic achievement, indicating that academic difficulties in COAs may be partly due to impaired motivation and organization.

Achievement↗
PubMed1998

A microscopical study of wound repair in the human placenta.

In order to fulfill its many functions as the selective interface between maternal and fetal circulations it is imperative that the human placenta remains intact and in good operational order. That damage of some sort occurs during its short but extremely active life seems inevitable given the dynamic environment in which the placenta exists, and evidence has accumulated that disruption is indeed a regular event. The implications of such damage, one could speculate, may impact on functions such as transport and hormone secretion as well as mutual protection against attack by maternal and fetal immune systems. Consequently, it would seem a theoretical necessity for discontinuities in the placenta surface to be repaired as soon as possible. We have used a combination of ex vivo observation, in vitro modelling, immunohistochemistry and correlative microscopy to provide evidence for a wound response in the placenta and to begin dissecting the detail of how this may operate. Evidence for small lesions caused by fusion and subsequent tearing of the syncytiotrophoblast in vivo, as well as plugging of such wounds by underlying cells is shown. We also identify a putative role for migratory cytotrophoblasts in the healing of larger scale injuries and demonstrate that certain molecules, common to wound repair in other tissues, appear to be involved in placenta repair also. Taken together these results clearly show that the human placenta is capable of a degree of self-maintenance by activating what appears to be an endogenous wound healing mechanism.

Chorionic Villi↗
PubMed1998

Bacterial products primarily mediate fibroblast inhibition in biomaterial infection.

PURPOSE: The stimulation of fibroblast growth is essential for the normal healing and tissue integration of biomaterials. The local elevation of proinflammatory mediators in infected perigraft fluid (PGF) may inhibit this growth. We sought to determine whether infected PGF inhibited fibroblast growth, and, if so, whether this was primarily dependent on the biomaterial, bacteria, or host. METHODS: In vivo Dacron or expandable polytetra-fluoroethylene (ePTFE) grafts, sterile or colonized with slime-producing (RP-62A, viable or formalin-killed) or nonslime-producing (RP-62NA) Staphylococcus epidermidis (1 x 10(7) CFU/cm2), were implanted in Swiss Webster mice, and the PGF was harvested at 7 and 28 days. Antibodies to tumor necrosis factor alpha, interleukin 1 alpha, interferon gamma (7 micrograms/day), and indomethacin (50 micrograms/day) were administered by microinfusion pumps for 7 days and the PGF was harvested. Inhibition of the proinflammatory mediators was confirmed by enzyme-linked immunosorbant assay. The nontreated, heat-treated, or trypsin-digested in vivo PGF was incubated with an in vitro [3H]thymidine murine fibroblast (ATCC CCL-12) proliferation assay. RESULTS: Fibroblast inhibition was significant at 7 and 28 days with infected PGF incubation compared with sterile and was not dependent on bacterial slime production or viability. Dacron sterile PGF did not significantly inhibit fibroblasts compared with control, whereas sterile ePTFE stimulated (P < 0.05) fibroblasts. Treatment of the PGF with proinflammatory cytokines, heat, and trypsin failed to reverse fibroblast inhibition in the infected state. CONCLUSION: Biomaterial infection is associated with fibroblast inhibition that is dependent primarily on bacterial products and not the host or biomaterial. Conservative intervention strategies for graft infection need to address the problem of poor healing as well as bacterial clearance.

Animals↗
PubMed1998

Changes in concentration, localization and activity of catalase within the human placenta during early gestation.

Using villous tissue from accurately dated gestational age placentae, this study identified significant changes in the protein concentration, enzyme activity and localization of catalase, an enzyme responsible for the intracellular metabolism of hydrogen peroxide, during the first and early second trimester of pregnancy. Enzyme activity was found to increase approximately threefold between weeks 6 and 17, with the greatest increase between 12 and 17 weeks. Immunostaining of tissue sections was supportive of these findings, demonstrating a progressively stronger signal between weeks 6 and 17. Immunostaining also demonstrated that the main cell types expressing catalase were the cytotrophoblast cells as well as a subset of the stromal cells. Between 13-17 weeks gestation, however, it was possible to detect catalase within the syncytiotrophoblast also, although with a much reduced intensity of staining. At the ultrastructural level, immunogold labelling of catalase clearly showed that staining was predominately compartmentalized within peroxisomes, although non-peroxisomal staining was also seen. Immunoreactivity also demonstrated, via morphological identification, that the stromal cells containing detectable levels of catalase were placental macrophages (Hofbauer cells). These results are in agreement with the proposal that the placenta exists in a physiologically low oxygen environment during the early part of gestation. In this environment oxidative activity of the sort resulting in the generation of hydrogen peroxide would presumably be suppressed, thereby limiting the requirement for catalase until oxygen tension begins to rise.

Catalase↗
PubMed1998

Susceptibility of human placental syncytiotrophoblastic mitochondria to oxygen-mediated damage in relation to gestational age.

When maintaining first trimester placental villi in organ culture under conventional normoxic conditions, we have observed widespread degeneration of the syncytiotrophoblast within 24 h despite excellent viability for the cytotrophoblastic and stromal cell types. Here we identify loss of mitochondrial activity as an early event in this process. In the light of proposals that the early part of gestation occurs in a low oxygen environment and also reported associations between mitochondrial disruption and oxidative stress, we cultured first trimester villi under low oxygen conditions (2.5%). Mitochondrial superoxide dismutase (MnSOD) localization and activity at different gestational ages were also determined. It was found that syncytiotrophoblastic and mitochondrial morphology improved, and mitochondrial activity was retained for 6 h and more if 8- to 10-week-old tissue was placed into a low oxygen environment immediately after removal from the uterus. The effect of oxygen concentration was less marked when using tissue of 14 weeks or more gestational age, which showed good survival and retention of mitochondrial activity under both low and ambient oxygen conditions. This correlated with our finding that placental MnSOD activity increased significantly between 8 and 14 weeks of gestation. Immunohistochemistry demonstrated that at 11 weeks; MnSOD was localized predominantly within the cytotrophoblast cells, whereas by 16 weeks it was found in the syncytiotrophoblast also. These results indicate an acute sensitivity of first trimester placenta syncytiotrophoblast to oxygen-mediated damage.

Culture Techniques↗
PubMed1997

Variations in expression of copper/zinc superoxide dismutase in villous trophoblast of the human placenta with gestational age.

This study investigated expression of the key antioxidant enzyme copper/zinc superoxide dismutase in the villous trophoblast of the human placenta at different gestational ages from 8 weeks (last menstrual period) to term. Immunostaining for the enzyme was observed in the cytotrophoblast cells at all stages. Staining was generally absent from the syncytiotrophoblast at 8 weeks, except for small isolated areas close to the basal surface. The size and location of these areas suggested they were the result of recent cytotrophoblastic fusion. By 10 weeks, examples were more frequent and diffuse staining throughout most of the syncytiotrophoblast was observed at 12 weeks. The intensity of the immunostaining within the syncytiotrophoblast continued to increase until 14 weeks, by which time it matched generally that within the cytotrophoblast cells. A similar pattern of staining was observed within term material. These results are entirely consistent with the hypothesis that the oxygen tension within the intervillous space is low throughout the first trimester of pregnancy. They support the idea that an effective maternal circulation to the human placenta is only established at the start of the second trimester.

Female↗
PubMed1997

Morphological analysis of degeneration and regeneration of syncytiotrophoblast in first trimester placental villi during organ culture.

We have recently shown using dansyl-L-lysine exclusion studies that the release of human chorionic gonadotrophin (HCG) in conjunction with L-lactate dehydrogenase (LDH) from first trimester villi during organ culture is symptomatic of syncytiotrophoblast degeneration. The purpose of this study was to examine chorionic villi at the ultrastructural level in order to determine events occurring during organ culture. The tissue was sampled after 0, 24, 48 and 120 h in culture and processed for electron microscopy. In addition to confirming the previously recorded syncytial degeneration, the electron micrographs showed clearly the generation of a new syncytiotrophoblast layer. The new layer, derived from differentiating cytotrophoblast cells, was largely formed by 48 h and was maintained for at least 120 h in culture. This study demonstrates a model which provides an opportunity to study the differentiation of cytotrophoblast cells whilst they retain their anatomical relationships within the villous structure.

Chorionic Villi↗
PubMed1996

An in vitro model for the study of wound healing in first trimester human placenta.

The placenta operates as a vital interface between the mother and fetus. In addition to facilitating fetal nourishment, it acts as a barrier both to potentially deleterious agents and to contact between their two immune systems. As a consequence, damage to the placenta, even on a relatively small scale, could be very dangerous to the fetus. Therefore, wound repair mechanisms are likely to be of great importance in ensuring that an intact placental barrier is re-established as soon as possible. By use of an in vitro method for injuring and subsequently culturing small pieces of first trimester villous tissue, we have observed a number of indications that a wound response is initiated. Pronounced expression of transforming growth factor-beta1, heavy infiltration of macrophages and late deposition of tenascin in the region of the wound all provide good evidence of some form of healing activity. Furthermore, we have noted that these indicators are suggestive of 'adult-type' rather than 'fetal-type' repair processes.

Female↗
PubMed1995

Life-span, T-cell responses, and incidence of lymphomas in congenic mice.

Survival, T-cell functions, and postmortem histopathology were studied in H-2 congenic strains of mice bearing H-2b, H-2k, and H-2d haplotypes. Males lived longer than females in all homozygous and heterozygous combinations except for H-2d homozygotes, which showed no differences between males and females. Association of heterozygosity with longer survival was observed only with H-2b/H-2b and H-2b/H-2d mice. Analysis using classification and regression trees (CART) showed that both males and females of H-2b homozygous and H-2k/H-2b mice had the shortest life-span of the strains studied. In histopathological analyses, lymphomas were noted to be more frequent in females, while hemangiosarcomas and hepatomas were more frequent in males. Lymphomas appeared earlier than hepatomas or hemangiosarcomas. The incidence of lymphomas was associated with the H-2 haplotype--e.g., H-2b homozygous mice had more lymphomas than did mice of the H-2d haplotype. More vigorous T-cell function was maintained with age (27 months) in H-2d, H-2b/H-2d, and H-2d/H-2k mice as compared with H-2b, H-2k, and H-2b/H-2k mice, which showed a decline of T-cell responses with age.

Animals↗
PubMed1995

Human chorionic gonadotrophin release and tissue viability in placental organ culture.

The use of human chorionic gonadotrophin (HCG) secretion as a measure of viability during the organ culture of human first trimester placental tissue has become a popular practice. It has been suggested that if cultured tissue is releasing large amounts of this protein hormone, there is a high level of viability. We have found, however, that the cytosolic enzyme L-lactate dehydrogenase is released into the culture supernatant in a similar daily pattern as HCG, suggesting that tissue disruption may be occurring, resulting in some of the observed hormone release. In addition, we have shown that the uptake of the fluorescent dye dansyl-L-lysine into the syncytium increases significantly from day 0 to day 4, suggesting a loss of syncytial membrane integrity. Electron micrographs show further evidence of the syncytial degeneration at the ultrastructural level, displaying extensive vacuolation and poor microvillous cover. In contrast to the degenerated state of the syncytiotrophoblast, a high level of bromodeoxyuridine incorporation is observed for cytotrophoblasts and, in particular, stromal cells up to 5 days in culture. Overall, the results suggest that the use of HCG release as a determinant of tissue viability in placental organ culture should be treated with a degree of caution.

Bromodeoxyuridine↗
PubMed1993

In vitro polymerization of oxidized tau into filaments.

Paired helical filaments (PHF) are abnormal neuronal polymers characteristic of Alzheimer's disease (AD). Although tau appears to be a major constituent of PHF, the mechanism for the polymerization of tau or its integration into PHF remains unknown. Here, we show that the oxidation of bovine tau in vitro induces an apparent dimerization of this protein and polymerization into filaments. These observations suggest that the oxidation of tau in vivo may contribute to the development of PHF in individuals with AD.

Actin Cytoskeleton↗
PubMed1992

Traits that influence longevity in mice: a second look.

Analysis of genetic interactions in the F2 of an intercross of (C57BL/6 x DBA/2) F1J revealed influences of genetic factors on life span. Females lived longer than males. Dilute brown females died sooner than females of other colors. H-2b/H-2b males died sooner than H-2b/H-2d or H-2d/H-2d males, except that among dilute brown males those of typeH-2b/H-2d died sooner. Cluster analysis suggested that male and female genotypes each fall into two groups, with female dilute brown mice having shorter lives than other females, and male H-2b/H-2b mice except dilute brown and dilute brown H-2b/H-2d mice having shorter lives than other males. The association of heterozygosity with life span was clearer in females than in males, yet the longest-lived female genotype was homozygous H-2d/H-2d, of dominant Black phenotype at the Brown locus of chromosome 4, and homozygous dd at the Dilute locus of chromosome 9. The shortest-lived females were dilute brown H-2b/H-2b. The longest-lived and shortest-lived male genotypes were dilute brown H-2d/H-2d and dilute brown H-2b/H-2d, respectively. Although histological findings at postmortem differed between the sexes, there was no association of particular disorders with other genetic markers. The importance of H-2 in males was confirmed, but the allelic effects were perturbed, possibly by the absence of Sendai infection in this experiment. Overall our studies suggest that genetic influences on life span involve interactions between loci, and allelic interactions may change with viral infections or other environmental factors.

Animals↗
PubMed1992

Dietary aluminum selectively decreases MAP-2 in brains of developing and adult rats.

Administration of 0.3% aluminum in drinking water elevated serum aluminum concentrations 8-fold in rats. Further, chronic treatment with aluminum for 2-3 mon, in both developing and adult rats, significantly decreased the levels of MAP-2 in brain, as determined by quantitative immunoblot analysis. Aluminum treatment also decreased the level of brain spectrin, but only in the hippocampus of adult rats. These were selective effects, since the levels of tubulin, tau and the three proteins of the neurofilament triplet were unaltered. In the aluminum-treated adult rats MAP-2 levels were significantly decreased in the hippocampus and brainstem to 71% and 56% of control values, respectively. In developing rats, MAP-2 levels were significantly decreased in the cortex and brainstem (65 and 64% of control values, respectively) but not in the hippocampus. In support of these findings, immunohistochemical examination revealed that the intensity of hippocampal MAP-2 immunoreactivity was significantly decreased to 88% of control values with aluminum treatment in adult rats. To determine a possible mechanism by which MAP-2 levels are reduced, the effect of aluminum on calpain-induced proteolysis of MAP-2 was examined in vitro. At the aluminum concentrations tested, there was no apparent effect on calpain-induced proteolysis of MAP-2. In the developing rats, aluminum administration significantly increased the hippocampal cyclic AMP concentration, as reported previously in adult aluminum-treated rats, and decreased the inositol 1,4,5-trisphosphate concentration. These results demonstrate that chronic oral aluminum administration to rats selectively decreases the levels of MAP-2 in specific brain regions independent of calpain proteolysis. This decrease may be associated with increased cyclic AMP and protein phosphorylation, and the impairment of cognition previously observed in this model of aluminum intoxication.

Administration, Oral↗