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Biomedical subjects

J M Roberts

Publications and source records attributed to J M Roberts.

At least 19 recordsLinked to original sources
PubMed1992

Formation and activation of a cyclin E-cdk2 complex during the G1 phase of the human cell cycle.

Human cyclin E, originally identified on the basis of its ability to function as a G1 cyclin in budding yeast, associated with a cell cycle-regulated protein kinase in human cells. The cyclin E-associated kinase activity peaked during G1, before the appearance of cyclin A, and was diminished during exit from the cell cycle after differentiation or serum withdrawal. The major cyclin E-associated kinase in human cells was Cdk2 (cyclin-dependent kinase 2). The abundance of the cyclin E protein and the cyclin E-Cdk2 complex was maximal in G1 cells. These results provide further evidence that in all eukaryotes assembly of a cyclin-Cdk complex is an important step in the biochemical pathway that controls cell proliferation during G1.

Animals↗
PubMed1992

Cell cycle regulated phosphorylation of RPA-32 occurs within the replication initiation complex.

The transition from G1 to S phase of the cell cycle may be regulated by modification of proteins which are essential for initiating DNA replication. One of the first events during initiation is to unwind the origin DNA and this requires a single-stranded DNA binding protein. RPA, a highly conserved multi-subunit single-stranded DNA binding protein, was first identified as a cellular protein necessary for the initiation of SV40 DNA replication. The 32 kDa subunit of RPA has been shown to be phosphorylated at the start of S phase. Using SV40 replication as a model, we have reproduced in vitro the S phase-dependent phosphorylation of RPA-32 and show that it occurs specifically within the replication initiation complex. Phosphorylated RPA-32 is predominantly associated with DNA. Phosphorylation is not a pre-requisite for association with DNA, but occurs after RPA binds to single-stranded DNA formed at the origin during the initiation phase. The protein kinase(s) which phosphorylates RPA-32 is present at all stages of the cell cycle but RPA-32 does not bind to the SV40 origin or become phosphorylated in extracts from G1 cells. Therefore, the cell cycle-dependent phosphorylation of RPA-32 may be regulated by its binding to single-stranded origin DNA during replication initiation.

Animals↗
PubMed1992

The concentration of estrogen receptors in rabbit uterine myocytes decreases in culture.

OBJECTIVE: The purpose of this study was to determine whether the concentration of estrogen receptors in cultured myocytes is preserved after dispersion. STUDY DESIGN: Primary myocytes were prepared from rabbit myometrium by collagenase dispersion after removing the endometrium and were isolated with Percoll density gradients. The cells were assayed for estrogen receptor concentration at intervals after dispersion by means of a whole-cell binding assay. Unpaired t test was used for comparisons. RESULTS: The concentration of estrogen receptors on the first day after dispersion was 12,058 +/- 1096 sites per cell (mean +/- SEM) and decreased to 4389 +/- 1223 site per cell within 9 to 14 days after dispersion (63% decline, p < 0.001). A similar decrease was observed when 2 nmol/L estradiol was present in the medium. CONCLUSION: The concentration of estrogen receptors in isolated rabbit uterine myocytes decreases after dispersion. This may partly explain the difficulty of demonstrating in vitro estrogen effects on myocytes, which are well established in vivo.

Animals↗
PubMed1992

Circulating concentrations of fetal fibronectin do not reflect reduced trophoblastic invasion in preeclamptic pregnancies.

This prospective, nested, case-control study investigated whether maternal plasma fetal fibronectin reflects reduced trophoblastic invasion at 16 to 20 weeks in women who later have preeclampsia. Concentrations of fetal fibronectin were 8.7 +/- 2.6 micrograms/ml in women with preeclampsia and 8.1 +/- 2.5 micrograms/ml in matched controls (p greater than 0.5). These results are discussed.

Case-Control Studies↗
PubMed1992

Activation of the p34 CDC2 protein kinase at the start of S phase in the human cell cycle.

Using a protocol for selecting cells on the basis of both size and age (with respect to the preceding mitosis), we isolated highly synchronous human G1 cells. With this procedure, we demonstrated that the p34 CDC2 kinase was activated at the start of S phase. Cyclin A synthesis began at the same time, and activation of the p34 CDC2 kinase at the start of S phase was, at least in part, due to its association with cyclin A. Furthermore, cells synchronized in late G1 by exposure to the drug mimosine contain active cyclin A/p34 CDC2 kinase, indicating that p34 CDC2 activation can occur before DNA synthesis begins. Thus, the cyclin A/CDC2 complex, which previously has been shown to be sufficient to start SV40 DNA synthesis in vitro, assembles and is activated at the start of S phase in vivo.

Burkitt Lymphoma↗
PubMed1992

Diloxanide furoate for treating asymptomatic Entamoeba histolytica cyst passers: 14 years' experience in the United States.

Diloxanide furoate is used for treating asymptomatic or mildly symptomatic persons who are passing cysts of Entamoeba histolytica. The Centers for Disease Control (Atlanta) released this drug for 4,371 treatment courses from 1977 through 1990. Of the 2,815 report forms (64%) returned, 656 adverse effects were reported for 390 treatment courses (14%); they included flatulence (260), diarrhea or cramping (100), nausea (93), headache (17), disorientation or dizziness (9), and diplopia (4). During 1984-1990 uniform collection of data allowed more detailed analysis of toxicity and efficacy; fewer adverse effects were reported for persons aged 20 months to 10 years than for persons aged greater than 10 years (6 of 206 [3%] vs. 89 of 763 [12%], relative risk = 0.27, 95% confidence interval = 0.12 less than relative risk less than 0.61). Parasitological cures were achieved during 497 (86%) of the 575 treatment courses (52%) administered to asymptomatic persons who were passing cysts, who had received a full 10-day treatment course, and for whom results of a follow-up stool examination (greater than or equal to 14 days post-treatment) were available. Diloxanide furoate is safe and effective for treating asymptomatic persons who are passing E. histolytica cysts and may be particularly well tolerated in children.

Adolescent↗
PubMed1992

Kinetics of circulating human IgG4 after diethylcarbamazine and ivermectin treatment of bancroftian filariasis.

Patent filarial infections are associated with elevated levels of parasite-specific IgG4. This study investigated the shifts of filarial-specific human IgG and IgG4 antibodies after diethylcarbamazine and ivermectin treatment of bancroftian filariasis. Thirty adult Haitians were treated first with a 1-mg clearing dose of ivermectin and then with either one or two 200-micrograms/kg doses of ivermectin or with 12 daily 6-mg/kg doses of diethylcarbamazine. Posttreatment levels of antifilarial IgG4 were dependent on both treatment group and time of follow-up. IgG4 increased markedly to a maximum by day 30 in all treatment groups and then began to decrease; the greatest decrease was among diethylcarbamazine-treated patients. Posttreatment microfilaremia was inversely correlated with the decrease in IgG4; thus, shifts in IgG4 were associated with treatment response for all groups. Antifilarial IgG levels were not correlated with drug treatment and did not change to the same degree as did IgG4 responses.

Animals↗
PubMed1992

Effects of thyroid hormone on myocardial adrenergic beta-receptor responsiveness and function during late gestation.

To assess the effect of fetal hypothyroidism during late gestation on postnatal cardiovascular responsiveness, we measured heart rate and cardiac output responses to isoproterenol in newborn lambs. To evaluate the effect of such altered thyroid status on the development of beta-adrenergic signaling cascade, we measured myocardial beta-adrenergic receptor concentration and affinity, guanine nucleotide regulatory protein density, and adenylyl cyclase responsiveness. Twenty fetal lambs underwent either thyroidectomy and line placement or line placement alone at 128-130 d gestation. Five thyroidectomized and six control newborns were treated with isoproterenol, five thyroidectomized and four control newborns were killed upon delivery and tissue was obtained for biochemical studies, and four additional animals were delivered and killed at 126 d gestation and tissue was obtained for receptor analysis. Of the newborns treated with isoproterenol, the thyroidectomized lambs showed lower increase in heart rate and cardiac output compared with euthyroid newborns. Compared with the myocardium of normal newborns of similar gestation, the myocardium of the newborns who underwent fetal thyroidectomy failed to show the normal increase in beta-adrenoceptors accompanied by reduction in beta-adrenergic-stimulated adenylate cyclase activity. These results suggest that near term, the normal development of ovine fetus myocardial beta-adrenergic receptor is affected by thyroid hormones.

Adenylyl Cyclases↗
PubMed1992

Characterization of the beta-adrenergic receptor in isolated human fetal lung type II cells.

Functioning of the beta-adrenergic response system is important for successful transition of the neonate from fetal life to breathing air. We characterized the beta-adrenergic receptors on human fetal lung type II cells, the cell type responsible for many pulmonary responses sensitive to beta-adrenergic stimulation. Type II cells were isolated from human fetal lung explants, and membrane particulates prepared from these cells were used for radioligand binding studies. 125I-iodocyanopindolol, a specific beta-adrenergic antagonist, bound to a single class of saturable, high-affinity binding sites on type II cell membranes with a receptor concentration of 78 +/- 9 fmol receptor/mg membrane protein, a kd of 79 +/- 18 nM, and 958 +/- 120 receptors per cell. Binding was stereoselective with l-propranolol binding with higher affinity than the inactive d-isomer. The binding site had the characteristics of a beta 2-adrenergic receptor. The order of potency of beta-adrenergic agonists was isoproterenol greater than epinephrine much greater than norepinephrine. The beta 2-selective antagonist ICI 118,551 competed for a single class of high-affinity sites. Agonist binding affinity was reduced in the presence of guanyl nucleotides, consistent with receptors coupled to guanine nucleotide binding proteins. beta-Adrenergic agonists also stimulated adenylyl cyclase in these membrane preparations. 125I-iodocyanopindolol binding to membranes prepared from human fetal lung fibroblasts indicated fewer receptors (404 +/- 68) than were present on type II cells. Work by others has suggested a difference in lung function and lung beta-adrenergic receptor concentration between males and females.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclases↗
PubMed1992

Cyclic adenosine 3',5'-monophosphate increases beta-adrenergic receptor concentration in cultured human fetal lung explants and type II cells.

cAMP regulates the maturation of many biochemical processes that occur during normal lung development, including the changing levels of surfactant proteins and phospholipids. We examined the effect of cAMP on the beta-adrenergic receptor concentration in the developing human lung. Isobutylmethylxanthine, a cAMP phosphodiesterase inhibitor, increased both the tissue cAMP content and beta-adrenergic receptor concentration in treated explants above those in untreated explants. 8-Bromo-cAMP treatment also elevated the beta-adrenergic receptor concentration of lung explants compared to that in untreated controls. These data indicate the ability of elevated cAMP to increase the beta-adrenergic receptor concentration. Both lung cAMP and beta-adrenergic receptor concentrations increase spontaneously in culture. To test for a possible causal relationship, we cultured explants with protein kinase inhibitors. We found that H-8, a preferential inhibitor of the cAMP-dependent protein kinase [protein kinase-A (PKA)], but not H-7, which inhibits PKA and protein kinase-C with similar potency, blocked the spontaneous rise in beta-adrenergic receptor concentration in human fetal lung explants, indicating that PKA activity is required for this rise in beta-adrenergic receptor concentration. Type II cells isolated from cultured lung treated with H-8 had fewer beta-adrenergic receptors than cells isolated from untreated explants. These studies show that cAMP increases the beta-adrenergic receptor concentration in human fetal lung and specifically in type II cells through a PKA-dependent mechanism, consistent with a role for cAMP in beta-adrenergic receptor regulation during normal lung development.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1992

Increases in plasma atrial natriuretic peptide concentration antedate clinical evidence of preeclampsia.

Atrial natriuretic peptide (ANP) concentrations are typically elevated in hypervolemic states. However, ANP levels have been reported to be increased in the peripartum period in women with preeclampsia, a disorder characterized by central hypovolemia. We postulated that ANP levels are elevated in preeclamptic patients before clinically evident disease. ANP concentrations were determined in three groups: uncomplicated pregnancies, pregnancies complicated by preeclampsia, and non-pregnant reproductive-aged women. The former groups were matched for gestational age at plasma sampling and delivery. The plasma samples, obtained prospectively from each patient during the first, second, and third trimesters and within 72 h postpartum, were frozen before RIA. A significant gestational increase in ANP was noted in both groups of pregnant women, with third trimester levels exceeding first trimester levels (P less than 0.05). Consistent with previous reports, ANP levels were elevated in overtly preeclamptic patients vs. matched controls in the third trimester. The ANP concentration was also significantly increased during the second trimester in women destined to develop preeclampsia. Postpartum ANP values decreased in the preeclamptic group to approach the level in normal patients postpartum. Thus, it appears that the stimuli of ANP secretion differ in uncomplicated and preeclamptic patients. Moreover, an elevation of plasma ANP is detectable before the onset of clinical evidence of preeclampsia.

Atrial Natriuretic Factor↗
PubMed1992

Exposure of human amnion to amniotic fluid obtained before labor causes a decrease in chorion/decidual prostaglandin release.

Prostaglandin (PG) production by fetal membranes has been implicated in the initiation of human parturition, but its regulation is not well understood. We used an in vitro system to study paracrine control of term, fetal membrane PG production. Using a modified Ussing chamber, full thickness fetal membranes with attached decidua were sealed into a chamber so that each hemichamber was a compartment for either the fetal (amnion) or maternal (chorion/decidua) side. Released PGs from maternal and fetal sides were then measured after exposure of the amnion to either buffer or amniotic fluid. We found that basal release of PGs from both the fetal and maternal sides was 2- to 3-fold higher in membranes obtained after labor compared to those obtained before labor. When amnion obtained after labor was exposed to amniotic fluid, we found a 3- to 5-fold increase in the net release of PGE2 from the amnion; however, the maternal side showed an unexpected relative decrease in PGE2 and PGF2 alpha release. This was a paracrine effect, since direct exposure of chorion/decidua to amniotic fluid caused increased release of the PG precursor, arachidonic acid. Direct transfer of radiolabeled PG from fetal to maternal side was minimal.

Amnion↗
PubMed1992

Epidemiology of giardiasis in Wisconsin: increasing incidence of reported cases and unexplained seasonal trends.

Giardia lamblia is the most commonly reported enteric pathogen in Wisconsin. Since giardiasis became a notifiable disease, the annual number of cases reported to the Wisconsin Division of Health has increased more than 20-fold, from 2.2 cases per 100,000 population in 1981 to 49.1 cases per 100,000 population in 1988. To better understand the nature of this increasing trend, we reviewed records of G. lamblia infections reported to the Wisconsin Division of Health from 1981 to 1988. Although the increase in reported cases was a general phenomenon that was not limited to a few high-risk groups, the highest annual incidence and greatest increase occurred in children 1-4 years old; 34% of the cases in this age group occurred in children who attended day care centers. A remarkably consistent late summer (August) increase was observed across all demographic and risk groups, suggesting that G. lamblia may be more common in the environment during late summer, or that risk factors for transmission may differ during these months. Additional studies are needed to further explain the increasing incidence and seasonal nature of reported giardiasis and to identify opportunities for prevention.

Adolescent↗
PubMed1992

Electrical stimulation for idiopathic scoliosis.

To evaluate the effectiveness of lateral electrical spinal stimulation for idiopathic scoliosis, 87 patients treated with this modality were reviewed retrospectively. All patients had no prior treatment, had a documented progression of more than 5 degrees, and were skeletally immature. Forty-seven patients were compliant and followed until skeletal maturity or institution of other treatment. Fifty percent of patients with a high probability of progression required surgery. For compliant patients, 51% progressed 5 degrees or more and 36% progressed 10 degrees or more or required a change to another treatment modality. Statistical analysis demonstrated no significant difference in the probability of progression between this group of treated patients and previously published groups of untreated patients.

Adolescent↗
PubMed1992

Expression of the CD46 antigen, and absence of class I MHC antigen, on the human oocyte and preimplantation blastocyst.

Expression of CD46 and class I major histocompatibility complex (MHC) antigens by human oocytes and 6-8-day unhatched expanded preimplantation blastocysts has been studied by immunocytochemistry. The CD46 antigen, a cell surface complement regulatory protein, was expressed by unfertilized oocytes as well as strongly by both the inner cell mass and trophectoderm of preimplantation blastocysts. In contrast, class I MHC antigens were not usually expressed by either oocytes or blastocysts. These data support the concept that gametes and embryonic cells involved in fertilization and early implantation events, respectively, may be protected from immunological recognition or attack both by the lack of class I MHC antigens and by expression of the CD46 complement regulatory protein.

Antibodies, Monoclonal↗
PubMed1991

Human cyclin E, a new cyclin that interacts with two members of the CDC2 gene family.

A new human cyclin, named cyclin E, was isolated by complementation of a triple cln deletion in S. cerevisiae. Cyclin E showed genetic interactions with the CDC28 gene, suggesting that it functioned at START by interacting with the CDC28 protein. Two human genes were identified that could interact with cyclin E to perform START in yeast containing a cdc28 mutation. One was CDC2-HS, and the second was the human homolog of Xenopus CDK2. Cyclin E produced in E. coli bound and activated the CDC2 protein in extracts from human G1 cells, and antibodies against cyclin E immunoprecipitated a histone H1 kinase from HeLa cells. The interactions between cyclin E and CDC2, or CDK2, may be important at the G1 to S transition in human cells.

Amino Acid Sequence↗
PubMed1991

Survey of veterinarians' recommendations for treatment and control of intestinal parasites in dogs: public health implications.

A systematic, random sample of 450 small and mixed-animal practitioners was selected from the client list of a prominent veterinary pharmaceutical and animal health company. A telephone survey was conducted, using a standard questionnaire, to assess whether current veterinary practices concerning prophylaxis and treatment of canine roundworm (Toxocara canis) and hookworm (Ancylostoma spp) infections are adequate to prevent transmission to human beings. Analysis of results focused on 3 questions related to prevention: practitioner's frequency of client education regarding zoonotic potential of roundworms and hookworms, pup age at which veterinarian recommends first anthelmintic treatments, and proportion of veterinarians recommending prophylactic drug administration for pups and nursing bitches. Despite the proven association of household pet dogs and human toxocariasis, only a third (148/450) of veterinarian respondents routinely discussed the potential zoonotic hazards of canine roundworms with their clients. A total of 29% (130/450) of veterinarians surveyed either never discussed these potential hazards or discussed them only when asked by their clients. With regard to anthelmintic treatment practices, 31% (140/450) of veterinarians surveyed recommended that pups first be examined and treated for intestinal parasites within 4 weeks of age. Thirty-three percent (163/450) recommended first examination and deworming at 5 to 6 weeks of age, and 36% (163/450) suggested that it be done at or after 7 weeks of age. Less than half (208/450) of veterinarians administered anthelmintics prophylactically to at least some pups and dogs. Sixty-four percent (287/450) of respondents recommended routine testing and treatment of nursing bitches.(ABSTRACT TRUNCATED AT 250 WORDS)

Ancylostomiasis↗