The cutting edge of plastic surgery: how to sharpen your Humby knife.
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Carbon Compounds, Inorganic↗
Biomedical subjects
M J Duncan
Publications and source records attributed to M J Duncan.
A disk of scattered icy objects and the origin of Jupiter-family comets.
Orbital integrations carried out for 4 billion years produced a disk of scattered objects beyond the orbit of Neptune. Objects in this disk can be distinguished from Kuiper belt objects by a greater range of eccentricities and inclinations. This disk was formed in the simulations by encounters with Neptune during the early evolution of the outer solar system. After particles first encountered Neptune, the simulations show that about 1 percent of the particles survive in this disk for the age of the solar system. A disk currently containing as few as approximately 6 x 10(8) objects could supply all of the observed Jupiter-family comets. Two recently discovered objects, 1996 RQ20 and 1996 TL66, have orbital elements similar to those predicted for objects in this disk, suggesting that they are thus far the only members of this disk to be identified.
Evolution, Planetary↗
Discrete thalamic lesions attenuate winter adaptations and increase body weight.
The midline thalamus (e.g., the paraventricular thalamic nuclei and the reuniens nucleus) of Siberian hamsters and other mammals has been reported to contain specific binding sites for melatonin, a hormone that is essential for photoperiodically induced winter adaptations such as reproductive quiescence, loss of body weight, daily torpor, and the winter molt. The first experiment investigated whether the midline thalamus is necessary for these winter adaptations. Adult Siberian hamsters received discrete neurotoxic lesions of the paraventricular thalamic nuclei or the reuniens nucleus while under pentobarbital sodium-induced anesthesia. After recovery, the hamsters were monitored for winter adaptations while they were exposed to short photoperiods (10 h light/day) for 12 wk at 22 degrees C then for 60 days at 7 degrees C. Lesions of the reuniens nucleus, but not of the paraventricular thalamic nuclei, significantly inhibited short photoperiod-induced loss of body weight and tended to increase food consumption and decrease daily torpor. The second experiment showed that lesions of the reuniens nucleus increased body weight gain compared with that in controls during exposure to long photoperiods at 22 degrees C for 16 wk. In summary, these findings show that the reuniens nucleus is an important site for regulation of body weight and suggest that lesions of the reuniens nucleus may attenuate winter metabolic adaptations by causing an increase in body weight.
Acclimatization↗
Need a pizza meshed skin? A simple expansion device.
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Developing Countries↗
kappa-opioid receptor expression defines a phenotypically distinct subpopulation of astroglia: relationship to Ca2+ mobilization, development, and the antiproliferative effect of opioids.
To assess the role of kappa-opioid receptors in astrocyte development, the effect of kappa-agonists on the growth of astroglia derived from 1-2-day-old mouse cerebra was examined in vitro. kappa-Opioid receptor expression was assessed immunocytochemically (using KA8 and KOR1 antibodies), as well as functionally by examining the effect of kappa-receptor activation on intracellular calcium ([Ca2+]i) homeostasis and DNA synthesis. On days 6-7, as many as 50% of the astrocytes displayed kappa-receptor (KA8) immunoreactivity or exhibited increases in [Ca2+]i in response to kappa-agonist treatment (U69,593 or U50,488H). Exposure to U69,593 (100 nM) for 72 h caused a significant reduction in number and proportion of glial fibrillary acidic protein-immunoreactive astrocytes incorporating bromodeoxyuridine (BrdU) that could be prevented by co-administering the kappa-antagonist, nor-binaltorphimine (300 nM). In contrast, on day 14, only 5 or 14%, respectively, of the astrocytes were kappa-opioid receptor (KA8) immunoreactive or displayed functional increases in [Ca2+]i. Furthermore, U69,593 (100 nM) treatment failed to inhibit BrdU incorporation at 9 days in vitro. Experimental manipulations showed that kappa-receptor activation increases astroglial [Ca2+]i both through influx via L-type channels and through mobilization of intracellular stores (which is an important Ca2+ signaling pathway in cell division). Collectively, these results indicate that a subpopulation of developing astrocytes express kappa-opioid receptors in vitro, and suggest that the activation of kappa-receptors mobilizes [Ca2+]i and inhibits cell proliferation. Moreover, the proportion of astrocytes expressing kappa-receptors was greatest during a period of rapid cell growth suggesting that they are preferentially expressed by proliferating astrocytes.
3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
Porphyromonas gingivalis genes isolated by screening for epithelial cell attachment.
Porphyromonas gingivalis is associated with chronic and severe periodontitis in adults. P. gingivalis and the other periodontal pathogens colonize and interact with gingival epithelial cells, but the genes and molecular mechanisms involved are unknown. To dissect the first steps in these interactions, a P. gingivalis expression library was screened for clones which bound human oral epithelial cells. Insert DNA from the recombinant clones did not contain homology to the P. gingivalis fimA gene, encoding fimbrillin, the subunit protein of fimbriae, but showed various degrees of homology to certain cysteine protease-hemagglutinin genes. The DNA sequence of one insert revealed three putative open reading frames which appeared to be in an operon. The relationship between P. gingivalis attachment to epithelial cells and the activities identified by the screen is discussed.
Amino Acid Sequence↗
Photoperiodic exposure and time of day modulate the expression of arginine vasopressin mRNA and vasoactive intestinal peptide mRNA in the suprachiasmatic nuclei of Siberian hamsters.
In hamsters, changes in ambient photoperiod lead to alterations in the circadian rhythm of pineal melatonin secretion and subsequent changes in reproductive function. The present study examined whether photoperiod also alters 24-h rhythms in neuropeptide mRNA levels in the SCN of Siberian hamsters. In situ hybridization and quantitative autoradiography were used to assess messenger RNA levels for vasopressin (AVP) and vasoactive intestinal peptide (VIP) in the SCN of hamsters sacrificed at six times of day following exposure to long (16 h light/day) or short (10 h light/day) photoperiod for 2 weeks. Both AVP mRNA and VIP mRNA in the SCN were significantly affected by time of day and photoperiodic exposure. The 24-h profiles of AVP mRNA and VIP mRNA showed different relationships to the light: dark cycle, suggesting that these profiles are differentially regulated. In general, short photoperiod tended to suppress AVP mRNA and VIP mRNA in the SCN; this effect on AVP mRNA was significant at two times of day. These results complement and extend previous findings of 24-h h profiles in neuropeptide mRNA expression in the rat SCN by showing that these 24-h profiles are also characteristic of the Siberian hamster SCN and that they can be modulated by photoperiod.
Animals↗
Photoperiodic effects on puberty and specific 2-[125I]iodomelatonin binding sites in Siberian hamsters.
When juvenile male Siberian hamsters are transferred from a long photoperiod to a short photoperiod, sexual maturation is greatly delayed by a pineal-dependent process. We hypothesized that the eventual onset of puberty during short photoperiod exposure may be caused by a loss of receptors for the pineal hormone, melatonin. This study quantitated specific 2-[125I]iodomelatonin binding sites in the suprachiasmatic nuclei and pars tuberalis of Siberian hamsters exposed to short photoperiod (10 h light per day) for either 12 or 30 weeks and in hamsters exposed to long photoperiod (16 h light per day) for the same time intervals. Photoperiodic exposure significantly affected testes weight. The hamsters exposed to long photoperiod for either 12 or 30 weeks had mean tests weights > 700 mg, in contrast to hamsters in short photoperiod for 12 weeks (mean testes weights < 30 mg) or 30 weeks (mean testes weights approximately 350 mg). The affinity of specific 2-[125I]iodomelatonin binding sites in both regions was significantly lower in hamsters exposed to short photoperiod as compared to hamster exposed to long photoperiod, at either 12 or 30 weeks. In contrast, there were no effects of photoperiod or duration of exposure on the density of specific 2-[125I]iodomelatonin binding sites in either the suprachiasmatic nuclei or the pars tuberalis. Furthermore, a change in the affinity of the specific 2-[125I]iodomelatonin binding sites in the suprachiasmatic nuclei was observed between the hamsters housed in short photoperiod for 12 weeks (sexually immature) and the hamsters housedin short photoperiod for 30 weeks (undergoing puberty).(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Porphyromonas gingivalis may multiply and advance within stratified human junctional epithelium in vitro.
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Adult↗
Effects of aging on photoperiodic responsiveness and specific 2-[125I]-iodomelatonin binding sites in the pars tuberalis and suprachiasmatic nuclei of Siberian hamsters.
Siberian hamsters undergo reproductive quiescence during exposure to a short day photoperiod, but this response appears to diminish with age. This study investigated whether age-related losses in photoperiodic responsiveness may be related to decreases in specific 2-[125I]-iodomelatonin binding sites in the suprachiasmatic nuclei or pars tuberalis. Adult male Siberian hamsters (group 1: 3-6 months of age and group 2: 9-12 months of age) were exposed to short photoperiod (10 hr of light/day) for 10 weeks. Profound testicular regression was evident in the majority of the hamsters in group 1, but in only a few of the hamsters in group 2. There were no significant differences between the groups in the density or affinity of the specific 2-[125I]-iodomelatonin binding sites in either the suprachiasmatic nuclei or the pars tuberalis. These findings suggest that the failure of older hamsters to respond to short photoperiod is not caused by a loss of specific 2-[125I]-iodomelatonin binding sites in the suprachiasmatic nuclei or pars tuberalis.
Aging↗
Investigations of the regulation of specific 2-[125I]iodomelatonin binding sites in Siberian hamsters by endogenous and exogenous melatonin.
These studies used quantitative in vitro autoradiography to investigate whether endogenous or exogenous melatonin modulate specific 2-[125I]iodomelatonin binding sites in the pars tuberalis or suprachiasmatic nuclei of Siberian hamsters. Saturation analyses were conducted on tissue sections from hamsters that were pinealectomized, exposed to constant illumination (72 h), or injected with melatonin, and from hamsters that were treated as controls. High affinity (Kd approximately 20-75 pM) specific 2-[125I]iodomelatonin binding sites were detected in the suprachiasmatic nuclei and pars tuberalis of all animals. Neither pinealectomy nor constant illumination significantly affected either the affinity or the density of the specific 2-[125I]iodomelatonin binding sites in either region. Melatonin injections led to a decrease in specific 2-[125I]iodomelatonin binding to the pars tuberalis at 3-4 h after the last injection. However, washing the sections before incubation with 2-[125I]iodomelatonin eliminated this effect, suggesting that melatonin was occupying the binding sites rather than decreasing their actual number. Furthermore, when hamsters were sacrificed 18 h after the last melatonin injection, no effect of melatonin on either the affinity or density of specific 2-[125I]iodomelatonin sites was observed. These data suggest that 2-[125I]iodomelatonin binding sites in Siberian hamsters are not regulated by changes in circulating melatonin levels.
Animals↗
Developmental appearance and age related changes in specific 2-[125I]iodomelatonin binding sites in the suprachiasmatic nuclei of female Syrian hamsters.
In Syrian hamsters, the circadian timing system is sensitive to melatonin during gestation but is not responsive in the adult. In order to further understand this developmental change in melatonin responsiveness, in vitro autoradiography was used to assess the presence of specific 2-[125I]iodomelatonin binding sites in the suprachiasmatic nuclei of female hamsters of selected embryonic (E) and postnatal (PN) ages (e.g. E13, E14, E15, PN1, PN2, PN12, PN25, PN112-133). Specific 2-[125I]iodomelatonin binding sites were seen in the suprachiasmatic nuclei of some of the E14 hamsters and all the perinatal hamsters (E15, PN1 and PN2) but not in older hamsters. In contrast, specific 2-[125I]iodomelatonin binding sites were seen in the pars tuberalis of all hamsters (with the exception of one), regardless of age. The transient expression of specific 2-[125I]iodomelatonin binding sites in the suprachiasmatic nuclei suggests that melatonin may have some special functions restricted to early development. The specific 2-[125I]iodomelatonin binding sites in the embryonic suprachiasmatic nuclei may represent the substrate for maternal melatonin to set the phase of the developing circadian timing system.
Aging↗
Characterization of hemolytic bacteria in subgingival plaque.
Three-quarters of the patients with periodontal diseases surveyed in this study had one or more distinct types of hemolytic bacteria in their subgingival plaque. Twelve different species of bacteria were identified, belonging to five genera (Actinomyces, Streptococcus, Staphylococcus, Prevotella, and Actinobacillus). Nine hemolytic isolates, consisting of four Prevotella denticola strains, two Actinomyces naeslundii genospecies 2 strains, and one each of P. melaninogenica, Streptococcus constellatus, and A. naeslundii genospecies 1 strains were characterized. Incorporation of pronase into blood agar medium inhibited hemolysis by all of the isolates, suggesting a proteinaceous component for each of their hemolysins. With one exception, hemolysin production appeared to be regulated by the concentration of environmental iron: exogenous hemin was found to inhibit hemolysin production, and the iron scavenging compound, 2,2'- dipyridyl, was found to promote hemolysin production by all of the strains except for the S. constellatus isolate. Genomic libraries of each of the hemolytic plaque isolates were prepared in Escherichia coli using pBR322. Hemolytic clones were isolated on blood agar medium containing ampicillin at frequencies ranging from 1-6.7 x 10(-4). Extensive restriction mapping revealed regions of homology in the case of clones derived from three P. denticola strains isolated from the same subjects. Two of the P. denticola-derived clones were virtually identical throughout the entrety of their > 5 Kb inserts. The clone derived from the third strain showed good homology to the other two within a 1.3 Kb region, but the flanking DNA showed no homology even though all three P. denticola isolates were shown to be clonally related by ribotyping.(ABSTRACT TRUNCATED AT 250 WORDS)
Actinomyces↗
Interactions of Porphyromonas gingivalis with epithelial cells.
The invasion of gingival epithelial cells by certain pathogenic periodontal bacteria may account for their presence within diseased gingival tissue. To dissect the initial steps of a potential invasion pathway for the periodontal pathogen Porphyromonas gingivalis, laboratory and clinical bacterial isolates were tested for their interactions with a human oral epithelial cell line (KB). Several P. gingivalis strains immobilized on filters could bind oral epithelial cells. Quantitative adherence assays supported these results. The invasion of epithelial cells by P. gingivalis 33277 was measured by assay and confirmed by transmission electron microscopy. These preliminary results demonstrate that certain P. gingivalis strains are capable of internalization by human oral epithelial cells in vitro.
Bacterial Adhesion↗
Autoradiographic localization of binding sites for 2-[125I]iodomelatonin in the pars tuberalis of the western spotted skunk (Spilogale putorius latifrons).
Pineal melatonin secretion mediates photoperiodic regulation of implantation of blastocysts in the female western spotted skunk. Autoradiography studies demonstrated that high affinity 2-[125I]iodomelatonin binding sites are present in the pars tuberalis but not in any other hypophyseal region or in the hypothalamus or thalamus of this species. This restricted localization of 2-[125I]iodomelatonin binding, which was characteristic of spotted skunks sacrificed at various times during their annual reproductive cycles, suggests that the pars tuberalis mediates photoperiodic responses in this species.
Animals↗
DNA sequence and in vitro mutagenesis of the gene encoding the fructose-1,6-diphosphate-dependent L-(+)-lactate dehydrogenase of Streptococcus mutans.
Previously, the fructose-1,6-diphosphate-dependent L-(+)-lactate dehydrogenase gene of Streptococcus mutans JH1000 was cloned into Escherichia coli (J. D. Hillman, M. J. Duncan, and K. P. Stashenko, Infect. Immun. 58:1290-1295, 1990). In the present study, the nucleotide sequence of 1.29 kb of S. mutans DNA which contained the promoter and protein-coding region of the gene was determined. In vitro disruption of the gene was achieved by deletion of the promoter and a major portion of the protein-coding sequence. Subsequently, a tetracycline resistance gene from S. mutans was inserted at the deletion site as a marker for selection. In addition, evidence from Southern hybridization showed that S. mutans JH1000 contained a single copy of the lactate dehydrogenase gene.
Amino Acid Sequence↗
Expression of the Chlamydia trachomatis major outer membrane protein-encoding gene in Escherichia coli: role of the 3' end in mRNA stability.
The major outer membrane protein (MOMP)-encoding gene (omp1) of Chlamydia trachomatis has been cloned into Escherichia coli and partially sequenced. This recombinant gene expresses a full-length 40-kDa product, which is recognized by a monoclonal antibody directed against the species-specific epitope of MOMP. The recombinant omp1 is expressed in either insertion orientation, indicating that it utilizes its own promoter system. The endogenous omp1 promoter possesses a relatively low activity despite the high level of MOMP expression. Deletion of a 520-bp fragment at the 3' end encoding 39 amino acids (aa) at the C terminus and the remainder of the noncoding region leads to a significant decrease in mRNA stability and loss of protein synthesis. When the MOMP-encoding plasmid was introduced into E. coli minicells, it expressed 40- and 43-kDa proteins; however, inhibition of post-translational processing by ethanol revealed only a 43-kDa protein. These data indicate that the unprocessed omp1 gene product contains a 22-aa leader sequence which is cleaved during translocation to the outer membrane, to yield a processed 40-kDa protein. The recombinant MOMP was localized to the outer membrane E. coli fraction, comparable to the location of the native C. trachomatis protein.
Amino Acid Sequence↗
Effects of melatonin agonists and antagonists on reproduction and body weight in the Siberian hamster.
This study examined whether melatonin-induced inhibition of testicular weight and body weight in vivo could be antagonized by luzindole, a competitive melatonin receptor antagonist, or methysergide, a competitive serotonin receptor antagonist. Adult male Siberian hamsters were exposed to a long photoperiod (16L:8D) and given daily injections of drugs 3 h before lights off for 7 weeks. Hamsters treated with melatonin (0.375 mg/kg) exhibited testicular regression and loss of body weight. These effects were also marked in hamsters treated concomitantly with melatonin (0.375 mg/kg) and luzindole (10 mg/kg). In other studies, chronic injections of luzindole (30 mg/kg) to juvenile hamster failed to antagonize testicular regression induced by either melatonin injections or exposure to a short day photoperiod (12L: 12D). In contrast, concommitant injections of methysergide (6.25 mg/kg) and melatonin attenuated testicular regression and loss of body weight. When administered alone, neither luzindole nor methysergide affected testicular weight or body weight, whereas chronic injections of 5-methyoxyluzindole (10 mg/kg) mimicked the inhibitory effects of melatonin. 5-Methoxyluzindole inhibits 2-[125I]-iodomelatonin binding to median eminence/pars tuberalis membranes with an affinity similar to that of melatonin. Luzindole shows lower affinity for the inhibition of 2-[125I]-iodomelatonin binding than melatonin, which may explain why luzindole is not an effective melatonin receptor antagonist when administered in vivo. Methysergide, which has a very low affinity for inhibition of 2-[125I]-iodomelatonin binding, probably inhibits the effects of melatonin by blocking serotonergic neurotransmission.
Animals↗