Biomedical subjects

J J Moore

Publications and source records attributed to J J Moore.

At least 37 records · Page 2Linked to original sources

PubMed1994

Kin selection, social structure, gene flow, and the evolution of chimpanzees.

Hypotheses about chimpanzee social behavior, phylogeography, and evolution were evaluated by noninvasive genotyping of free-ranging individuals from 20 African sites. Degrees of relatedness among individuals in one community were inferred from allele-sharing at eight nuclear simple sequence repeat (SSR) loci. Males are related on the order of half-siblings, and homozygosity is significantly increased at several SSR loci compared to Hardy-Weinberg expectations. These data support the kin-selection hypothesis for the evolution of cooperation among males. Sequence variation patterns at two mitochondrial loci indicate historically high long-distance gene flow and clarify the relationships among three allopatric subspecies. The unexpectedly large genetic distance between the western subspecies, Pan troglodytes verus, and the other two subspecies suggests a divergence time of about 1.58 million years. This result, if confirmed at nuclear loci and supported by eco-behavioral data, implies that P. t. verus should be elevated to full species rank.

Africa↗

PubMed1994

Augmentation of antigen-specific lymphoproliferative responses in vitro by biological response modifiers.

The detection of antigen-specific T cell responsiveness, particularly of resting memory lymphocytes, in cultures of peripheral blood mononuclear cells (PBMC) may be hampered by a less than optimal antigen presentation in vitro. Augmented sensitivity of the test system may be achieved by the addition of reagents with a beneficial effect on lymphocyte and antigen-presenting cell (APC) functions. In this study the effect of several biological response modifiers on antigen-specific T cell proliferation was determined, using nickel sulphate and tetanus toxoid as test antigens. IL-1 alpha (100 U/ml), interferon-gamma (IFN-gamma) (10 U/ml), and indomethacin (2 microM) were found to significantly enhance nickel-induced proliferation in PBMC cultures from nickel-hypersensitive donors (n = 6). Tetanus-induced proliferation (n = 5) was similarly enhanced, both by the above supplements and by the addition of polyethylene glycol (PEG) or a neuraminidase treatment of the PBMC before culture. The addition to PBMC cultures of a combination of IL-1 alpha (30 U/ml), IFN-gamma (10 U/ml), and indomethacin (2 microM) is recommended to specifically enhance antigen-induced lymphoproliferative signals.

Antigen Presentation↗

PubMed1994

Paternity exclusion in a community of wild chimpanzees using hypervariable simple sequence repeats.

We report the use of hypervariable simple sequence repeat (SSR) nuclear loci to study paternity in a community of wild chimpanzees (Pan troglodytes schweinfurthii) in Gombe National Park, Tanzania. All 43 living members of a habituated community were sampled and 35 were genotyped at 8 SSR loci using DNA amplified from hair. Paternity exclusions were performed for 25 chimpanzees including 10 for whom the mother was also genotyped. In each case 12-20 males were potential fathers based on their age and/or direct observation of sexual behaviour. 179 tests involving potential father/offspring combinations were performed. In four cases the data permit the probable identification of the previously undetermined father; these are the first such determinations for free-ranging chimpanzees, and the first based on non-invasive sampling. In another four cases we were able to exclude all but two to five potential fathers, and in the remaining cases we were able to exclude all living males. For molecular ecologists SSR genotype databases offer important advantages over currently popular minisatellite DNA finger-printing: they can be analysed unequivocally using traditional population genetics techniques and they can be expanded through time and space by other researchers.

Alleles↗

PubMed1993

Protein kinase A activators inhibit agonist induced prostaglandin production in human amnion.

Prostaglandin (PG) production by human amnion has been postulated to have a role in the onset of labor. Previous work by ourselves and others has demonstrated that oxytocin, phorbol esters and epidermal growth factor (EGF) increase PGE2 production in human amnion cells by activation of the Phospholipase C/Protein Kinase C (PKC) cascade system. The present study was undertaken to determine the effect of prior activation of the Adenylate Cyclase cascade system upon subsequent stimulation of PGE2 production by oxytocin, phorbol 12-myristate-13-acetate (PMA) or EGF in amnion cells and membrane discs. Isoproterenol, forskolin and dibutyryl cyclic adenosine monophosphate (dbcAMP) were utilized to activate the Adenylate Cyclase system at the receptor, enzyme and second messenger level. In control amnion cells, oxytocin, PMA and EGF each provoked dose dependent increases in PGE2 production. In cells preincubated with dbcAMP, forskolin or isoproterenol, agonist stimulated PGE2 production was markedly (50-90%) inhibited (p < 0.01). Inhibition was dose dependent upon preincubator concentrations. Maximal inhibition by adenylate cyclase activators occurred with 2-4 h of preincubation. In membrane discs, forskolin preincubation also inhibited oxytocin, PMA and EGF stimulation of PGE2 production. Activation of the Adenylate Cyclase system in human amnion cells or membrane discs inhibits the subsequent action of potent stimulators of PGE2 production in human amnion.

Adenylyl Cyclases↗

PubMed1993

Ritodrine: a beta-adrenergic receptor antagonist in human amnion.

OBJECTIVE: Human amnion is important in the initiation of labor. Ritodrine, when administered as a tocolytic, is found unchanged in amniotic fluid. We characterized effects of ritodrine binding to beta-adrenergic receptors in amnion and amniocytes. STUDY DESIGN: Iodine 125-iodopindolol, beta-adrenergic receptor agonists, and beta-adrenergic receptor antagonists were used to describe binding characteristics. Experiments were designed with and without isoproterenol and ritodrine to study intracellular cyclic adenosine 3'5'-monophosphate and prostaglandin E2 release. RESULTS: Scatchard analysis revealed a single class of saturable binding sites, with maximum binding capacity of 70.0 +/- 17.2 fmol/mg protein (n = 12) and with high-affinity dissociation constant of 458.9 +/- 72.1 pmol/L. Agonists and antagonists competed for the 125I-iodopindolol binding site consistent with a beta 2-adrenergic receptor. Hill coefficients were 0.6 to 0.8 for agonist competition and 1.0 for antagonist competition and ritodrine. Stimulation with isoproterenol resulted in dose-dependent increases in cyclic adenosine 3'5'-monophosphate and prostaglandin E2. Ritodrine failed to stimulate cyclic adenosine 3'5'-monophosphate and inhibited isoproterenol-stimulated cyclic adenosine 3'5'-monophosphate and prostaglandin E2 production. CONCLUSION: In human amnion binding of ritodrine to beta 2-adrenergic receptors and lack of ritodrine-mediated postreceptor effects are characteristic of a beta 2-adrenergic antagonist.

Amnion↗

PubMed1993

Human fetal membranes inhibit spontaneous uterine contractions.

Fetal membranes are postulated to play a role in paracrine signaling during the initiation of labor in women. We developed a dual chamber-fetal membrane-uterine muscle model to study the effect of human fetal membranes on spontaneous uterine contractions. In this model, full-thickness fetal membranes (amnion, chorion, and maternal decidua) are sealed into a Plexiglass chamber. The membranes partition the chamber into a maternal and fetal compartment. Chorion and decidua face the maternal side, and amnion faces the fetal side. An estrogenized rat uterine muscle strip is anchored into the maternal side as a bioassay to measure effects of fetal membranes on uterine contractions. Fetal membranes cause a 40% decrease in uterine contractions compared to basal condition (no membranes). Inhibition is reversible after removal of the membranes. The inhibition is specific to the chorion/decidual side because reversal of membranes with amnion toward the muscle did not show inhibition. Uterine contractions did not change over time in control chambers in which Parafilm substituted for membranes. A model for studying paracrine regulation of uterine contractions by human fetal membranes has been developed. The model provides evidence that fetal membranes inhibit uterine contractions. This inhibitory effect may contribute to uterine quiescence during pregnancy.

Animals↗

PubMed1993

Tolerance of sewage treatment plant microorganisms to mosquitocides.

Beneficial protozoa and rotifers collected from a wastewater treatment plant in Panama City, FL, were tested for tolerance to 11 commonly used mosquito larvicides and adulticides in the laboratory. The acute effects were assessed using selected concentrations of the adulticides fenthion, malathion, naled, permethrin, and resmethrin; and the larvicides Bacillus thuringiensis israelensis, Bacillus sphaericus, diflubenzuron, larviciding oil, methoprene, and temephos for the following microorganism taxa: ameoboids, flagellates, free-swimming ciliates, stalked ciliates, and rotifers.

Animals↗

PubMed1992

Identification of chimpanzee subspecies with DNA from hair and allele-specific probes.

We describe a non-invasive method of determining the subspecies identity of common chimpanzees (Pan troglodytes), based on subspecies-specific sequence differences in the mitochondrial genome. This procedure involves the extraction of DNA from hair, the amplification of a short (410 base pair (b.p.)) segment of the non-coding displacement loop (D-loop) by the polymerase chain reaction (PCR), and subspecies identification based on rapid allele-specific oligonucleotide (ASO) probe dot-blot typing. This approach will contribute to: (i) the colony-level management of captive chimpanzees by enabling managers to recognize hybrids between subspecies and minimize outbreeding depression; (ii) the recognition of inappropriately matched individuals in comparative behavioural and experimental studies; and (iii) forensic questions surrounding the origin of illegally traded animals.

Alleles↗

PubMed1992

Fatal toxoplasmosis in domestic rabbits in the USA.

Three rabbits from two sources died after an acute illness characterized by fever, lethargy and diarrhea in one rabbit and no clinical signs in two rabbits. The most striking lesion in all three rabbits was foci of necrosis of the spleen and liver associated with massive presence of multiplying Toxoplasma gondii tachyzoites. The diagnosis was confirmed by specific staining with anti-T. gondii serum in an avidin-biotin complex immunohistochemical stain.

Animals↗

PubMed1992

Human T lymphocyte cAMP-dependent protein kinase: subcellular distributions and activity ranges of type I and type II isozymes.

The role of the type I and type II protein kinase A isozymes in the regulation of human T lymphocyte immune effector functions has not been ascertained. To approach this question, we first characterized the distribution and enzyme activities of the type I and type II protein kinase A (PKA) isozymes in normal, human T lymphocytes. T cells possess both type I and type II isozymes with an activity ratio of 5.0:1 +/- 0.71 (mean +/- SD). The type I isozyme associates predominately with the plasma membrane whereas the type II isozyme localizes primarily to the cytosol. Analyses of isozyme activities demonstrated that T cells from approximately one-third of 16 healthy donors exhibited significantly higher type II isozyme activities (higher type II, type IIH) than the remaining donors (lower type II, type IIL) (mean = 605 +/- 75 pmol.min-1.mg protein-1, P less than 0.001). Scatchard analyses of [3H]cAMP binding in the cytosolic fraction demonstrated similar Kd values (type IIH, 1.1 x 10(-7) M; type IIL, 9.0 x 10(-8) M); however, the Bmax (maximal binding) of the type IIH was 400 fmol/mg protein compared to the Bmax of the type IIL of 126 fmol/mg protein. Scatchard analysis of [3H]cAMP binding to the type I isozyme associated with membrane fragments had a Kd of 5.6 x 10(-8) M and a Bmax of 283 fmol/mg protein. Eadie-Hofstee plots of type IIH and type IIL gave a Km and Vmax of 2.3 mg/ml and 1.5 nmol.mg-1.min-1, and 2.1 mg/ml and 1.6 nmol.mg-1.min-1, respectively. The 3.2-fold higher maximal binding of the type II isozyme in one-third of healthy donors may reflect a greater amount of isozyme protein. The compartmentalization of type I PKA isozyme to the plasma membrane and type II PKA isozyme to the cytosol may serve to localize the isozymes to their respective substrates in T lymphocytes.

Adult↗

PubMed1991

The effects of 5-fluorouridine, 5-fluorodeoxyuridine, and 5-fluorodeoxyuridine monophosphate on rabbit tenon's capsule fibroblasts in vitro.

Inhibition of rabbit subconjunctival fibroblast attachment and proliferation by 5-fluorouridine (FUR), 5-fluoro-2 deoxyuridine (FUdR), and 5-fluoro-2-deoxyuridine-5-monophosphate (FdUMP), was determined by 3H-adenosine uptake, cell counting, and colorimetric assays for the concentration range of 1000 to 0.0001 micrograms/ml over an 9 day period. The mean 50% inhibitory doses against proliferation were calculated for each assay. Rabbit fibroblast attachment was not inhibited at any drug concentration by either FUR, FUdR, or FdUMP. For rabbit fibroblast proliferation, FUR was found to be 10-100 fold more potent than FUdR and FdUMP. When comparing the human and rabbit cells, the unpaired t-test analysis showed no consistent statistical difference of the ID50s for FUR, FUdR or FdUMP. Rabbit ocular fibroblasts may be useful in modeling the proliferation of human ocular fibroblasts. These in vitro results may be useful for predicting optimal drug dosages for future in vivo testing of these drugs.

Adenosine↗

PubMed1991

Protein kinase-C activation is required for oxytocin-induced prostaglandin production in human amnion cells.

In previous work we reported that oxytocin activates phospholipase-C (PLC) and increases prostaglandin E2 (PGE2) release in amnion. Whether either of the consequences of activation of PLC by oxytocin, activation of protein kinase-C (PKC) or increases in intracellular calcium, directly results in the production of PGE2 is unknown. Phorbol esters (PMA) and epidermal growth factor (EGF) are also known to increase PGE2 release from amnion. In some tissues these agents are capable of activating the PLC postreceptor cascade system. This study was undertaken primarily to explore the mechanism of oxytocin-induced PGE2 production in amnion and secondarily to determine whether common aspects of PGE2 production by oxytocin, PMA, and EGF include activation of PLC or subsequent steps in this cascade followed by new mRNA/protein production. Involvement of PLC was assessed by inositol phosphate (IP1) turnover. IP1 turnover was increased by oxytocin (2.99 +/- 0.31-fold; P less than 0.01), but not by EGF or PMA. PMA inhibited oxytocin-provoked IP1 turnover (P less than 0.05). PKC involvement was initially evaluated with two PKC inhibitors, H7 and staurosporine. Each inhibited PGE2 production by oxytocin as well as that by PMA and EGF in a dose-dependent fashion. With H7, the IC50 for all agents was 5 microM; the IC50 for staurosporine was 2 nM for PMA and oxytocin and 5 nM for EGF. Agonist-induced PGE2 production was also assessed in cells in which PKC activity had been tachyphylaxed with a high concentration of PMA (400 ng/mL for 48 h). In such cells oxytocin and PMA no longer stimulated (P less than 0.001) PGE2 production, but EGF-stimulated PGE2 production was only slightly reduced. PKC involvement is, thus, implicated for oxytocin and PMA. Other enzymes that are inhibited by H7 and staurosporine are implicated in the production of PGE2 caused by EGF. Although tachyphylaxed cells produced no PGE2 with oxytocin, oxytocin increased intracellular calcium to levels higher than those seen in control cells (435 +/- 102 vs. 286 +/- 1.2) Actinomycin-D (P less than 0.001) and cycloheximide (P less than 0.05) inhibited PGE2 production caused by oxytocin, PMA, and EGF. PGE2 production by oxytocin in human amnion cells proceeds by activation of PKC, followed by new protein and mRNA production. Further, in cells without PKC, oxytocin-induced calcium transients do not increase PGE2. The ability of EGF to stimulate PGE2 in cells with no PKC activity also establishes that PKC activation is not a common intracellular step in the induction of PGE2 production by all agents.

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

PubMed1991

Evaluation of a total T3 assay on the Stratus fluorometric enzyme immunoassay system.

The Stratus total triiodothyronine (T3) immunoassay is an automated fluorometric enzyme immunoassay that utilizes a mouse monoclonal anti-T3 antibody preimmobilized onto glass fiber paper. The rate of formation of the enzyme product, as measured by front surface fluorometry, is inversely proportional to Total T3 concentration in the sample. The authors evaluated this method with respect to precision, sensitivity, interfering factors, and correlation with a radioimmunoassay. The overall, between-run, and within-run precision of the assay measured at three concentration levels for a total of 60 determinations each using immunoassay control materials, ranged from 2.5% to 14.3%. A total of 200 specimens, including 40 classified as hyperthyroid, and 38 classified as hypothyroid were analyzed in duplicate by the Stratus (STR) system and by a commercially available radioimmunoassay method. The coefficient of correlation obtained was 0.97. Icteric, hemolyzed, azotemic, and lipemic samples were included in the comparison and do not appear to have any interfering effect on the assay. The range of the assay is from 0.8 to 12 nmol/L. In summary, the Stratus Total T3 immunoassay offers the advantages of sensitivity, specificity, and automation with a throughput rate of 45 samples/h.

Evaluation Studies as Topic↗

PubMed1989

Neonatal resuscitation in community hospitals. A regional-based, team-oriented training program coordinated by the tertiary center.

Neonatal resuscitation in community hospitals is problematic because of the lack of on-site personnel with adequate training and experience. Even when efforts are made to transport high-risk mothers to tertiary care centers, the most complex resuscitations must occasionally be performed in all obstetric services. This dilemma can result in increased neonatal morbidity and mortality, and is reflected in the obstetric malpractice crisis. We present an approach to the problem in which a tertiary center actively intervenes to assist regional community hospitals to develop and implement resuscitation team capabilities. Key aspects of the system are training and implementation of hospital shift "Code Pink" teams composed of nonphysician professionals, a regional neonatal resuscitation protocol used by all participants, "mock code" resuscitation drills to maintain skills, and extensive quality assurance both at the local hospital and regional levels.

Clinical Protocols↗

PubMed1989

The Georgia family Q-sort: an observational measure of family functioning.

The development of a new observational measure of family functioning using a Q-sort methodology is described. The Q-sort is one way of obtaining observer measures of family that complements both microcoding and global ratings of family processes. Evidence for construct validity is presented along with a discussion of the advantages and disadvantages of the Q-sort approach to family observation.

Adaptation, Psychological↗

PubMed1989

Adenosine triphosphate activates the phospholipase-C cascade system in human amnion cells without increasing prostaglandin production.

Human amnion is hypothesized to be a target tissue for hormone messages from the fetus regarding labor. We have previously demonstrated prostaglandin E2 (PGE2) release in amnion after treatment with phorbol and oxytocin, but other potential agonists of the inositol phospholipid/protein kinase-C system have not been investigated. The effects of extracellular ATP on cytosolic calcium concentration [( Ca2+])i) inositol phosphate (IP) accumulation, and PGE2 production were studied in cultured human amnion cells. Intracellular free calcium [Ca2+]i was measured using the fluorescent dye fura-2. Addition of 0.01-30 microM ATP resulted in a [Ca2+]i transient which peaked within 15 sec and returned to baseline over 10 min. UTP (1 microM) was more effective than ATP (1 microM); [Ca2+]i levels rose from 233 to 2880 nM (UTP) and 2320 nM (ATP). A reduced effect was observed with other nucleotides in a rank order of agonist potency of ITP greater than CTP greater than ADP greater than GTP greater than TTP. No effect was seen with AMP, cAMP, or adenosine. This is consistent with P2 purinoceptors, as described in other tissues. ATP (100 microM) also dramatically increased IP accumulation. Inositol triphosphate, inositol bisphosphate, and inositol monophosphate were increased 7-, 9-, and 16-fold respectively. The agonist potency order of other nucleotides for IP accumulation was the same as that of [Ca2+]i. Pharmacological concentrations of ATP (1 mM) were required to increase PGE2 production. Many other nucleotides were equally effective at this concentration. ATP activates the phospholipase-C system in human amnion, as demonstrated by the increase in [Ca2+]i and inositol phosphates. The physiological significance of purinergic stimulation of this tissue remains unclear.

Adenosine Triphosphate↗

PubMed1988

Polyamines control human chorionic gonadotropin production in the JEG-3 choriocarcinoma cell.

The effect of inhibition of ornithine decarboxylase with difluoromethylornithine (DFMO) and the resultant lowering of polyamine levels upon human chorionic gonadotropin (hCG) production in JEG-3 choriocarcinoma cells was investigated. DFMO (10 mM) totally inhibited ornithine decarboxylase activity. In DFMO-treated cells, cellular spermidine concentrations fell to nondetectable levels (less than 1% of control values) within 24 h and spermine concentrations were reduced to 41.9% of controls over 6 days. DFMO caused a 70-80% inhibition of hCG production. Levels of mRNA for both the alpha and beta subunits of hCG were also inhibited relative to mRNA for tubulin. Exogenous putrescine normalized hCG production in a dose-dependent manner. Other diamines, including cadaverine, 1,3-diaminopropane, 1,6-diaminohexane, and 1,7-diaminoheptane, were ineffective in reestablishing hCG production in DFMO-treated cells. Dibutyryl cAMP (1 mM) stimulated hCG production and increased levels of mRNA for the alpha and beta subunit 5-40-fold in both DFMO-treated and control cells. Polyamines appear to have a fundamental role in hCG production in JEG-3 choriocarcinoma cells. However, dibutyryl cAMP can partially overcome or circumvent the requirement for polyamines in hCG biosynthesis.

Bucladesine↗

PubMed1988

Dexamethasone effects on ritodrine-induced changes in myometrial contractility and beta-adrenergic receptor function.

We have previously demonstrated in pregnant sheep that ritodrine infusion for 24 hours reduces myometrial beta-adrenergic receptor density and isoproterenol-stimulated adenylate cyclase activity. These receptor-associated changes were accompanied by an increasing inability of ritodrine to inhibit uterine contractility induced by a bolus of oxytocin. In the present study, we evaluated whether these ritodrine-induced effects could be altered by dexamethasone. Ten pregnant sheep at gestational ages of 92 to 130 days received ritodrine 2 micrograms/kg/min for 24 hours. Five animals also received dexamethasone 10 mg intravascularly twice during the ritodrine infusion. Before and at 4 and 24 hours of ritodrine infusion, the animals were given an identical dose of oxytocin as a bolus, and the area under the uterine pressure-time curve was quantified. Myometrial biopsy specimens were obtained before and after ritodrine infusion. Dexamethasone treatment prevented ritodrine-induced reductions in beta-adrenergic receptor density and isoproterenol-stimulated adenylate cyclase activity. Despite these receptor-associated effects, dexamethasone did not prevent the loss of tocolytic efficacy associated with prolonged ritodrine infusion.

Animals↗