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Biomedical subjects

D L Lewis

Publications and source records attributed to D L Lewis.

At least 55 records · Page 3Linked to original sources

Cross-infection risks associated with current procedures for using high-speed dental handpieces.

When a dye solution used to simulate patient material was either injected into high-speed dental handpiece (drill) waterlines or applied to the equipment externally, internal air turbine chambers became contaminated. These chambers served as a reservoir of the material, which was slowly dislodged by air expelled during subsequent handpiece operation and which was diluted by water spray used for cooling the drilling surface. Considering the fact that patient materials could reside in internal parts of the equipment that are not usually disinfected and that the material may be subsequently sprayed into cuts and abrasions in the oral cavity, the common approach to reprocessing handpieces (external wiping in combination with flushing) may pose unacceptably high risks to those individuals treated soon after infected patients. Therefore, unless reliable data on cross-infection frequencies are obtained and prove it unnecessary, heat-treating high-speed handpieces between each patient should be considered an essential component of standard procedures whenever universal precautions are practiced in dentistry.

Cross Infection↗

Infection control.

Explore the source record for details and available documents.

Dental High-Speed Equipment↗

Expression of an inwardly rectifying K+ channel from rat basophilic leukemia cell mRNA in Xenopus oocytes.

Rat basophilic leukemia cells (RBL-2H3) have previously been shown to contain a single type of voltage-activated channel, namely an inwardly rectifying K+ channel, under normal recording conditions. Thus, RBL-2H3 cells seemed like a logical source of mRNA for the expression cloning of inwardly rectifying K+ channels. Injection of mRNA isolated from RBL-2H3 cells into Xenopus oocytes resulted in the expression of an inward current which (1) activated at potentials negative to the K+ equilibrium potential (Ek), (2) decreased in slope conductance near Ek, (3) was dependent on [K+]o and (4) was blocked by external Ba2+ and Cs+. These properties were similar to those of the inwardly rectifying K+ current recorded from RBL-2H3 cells using whole-cell voltage clamp. Injection of size-fractionated mRNA into Xenopus oocytes revealed that the current was most strongly expressed from the fraction containing mRNA of approximately 4-5 kb. Expression of this channel represents a starting point for the expression cloning of a novel class of K+ channels.

Animals↗

Quantitative assessment of the effects of metals on microbial degradation of organic chemicals.

Biodegradation inhibition of a benchmark chemical, 2,4-dichloro-phenoxyacetic acid methyl ester (2,4-DME), was used to quantify the inhibitory effects of heavy metals on aerobic microbial degradation rates of organic chemicals. This procedure used lake sediments and aufwuchs (floating mats) collected in the field or from laboratory microcosms. Effects of CuCl2, HgCl2, ZnCl2, Cd(NO3)2, and Cr(NO3)3 at initial concentrations ranging from 0.3 microM to 73 mM (approximately 0.1 to 10,000 mg liter-1) were investigated. In general, such metallic compounds appeared to be considerably more inhibitory to the biodegradation of an organic chemical than high concentrations of microbially toxic organics studied previously. Effects of various metal concentrations were evaluated based on the following: (i) estimated MICs, (ii) concentrations that caused a significant effect on biodegradation parameters (both a greater than 10% decrease in Vmax and a greater than 10% increase in t1/2 for 2,4-DME degradation), and (iii) concentrations that caused biodegradation half-life doublings (HLDs). The MICs of metals in sediment were lowest for Zn2+ (0.10 microM) and highest for Cd2+ and Cu2+ (0.9 and 1.2 microM, respectively). The MICs of metals in aufwuchs were lowest for Hg2+ (0.01 microM), intermediate for Cu2+ and Zn2+ (0.42 and 0.62 microM, respectively), and highest for Cr3+ and Cd2+ (3.4 and 5.6 microM, respectively). Compared with Cu2+ on aufwuchs, 70 times more Zn2+, 250 times more Cr3+, and 1,000 times more Cd2+ was required to significantly affect aufwuchs biodegradation rate parameters and coefficients (Vmax and t1/2). Aufwuchs was significantly affected by the lowest Hg2+ concentration tested (5 microM).(ABSTRACT TRUNCATED AT 250 WORDS)

2,4-Dichlorophenoxyacetic Acid↗

Functional expression of dihydropyridine-insensitive calcium channels during PC12 cell differentiation by nerve growth factor (NGF), oncogenic ras, or src tyrosine kinase.

1. Recombinant retroviruses were used to introduce a temperature-sensitive v-src gene and oncogenic c-Ha-ras into PC12 cells, and stable cell lines expressing these genes were established. 2. As previously reported, expression of v-src (Alema et al., 1985) or c-Ha-ras (Noda et al., 1985) in PC12 cells results in neurite outgrowth resembling that induced by NGF. We report here that v-src but not oncogenic c-Ha-ras induces a stable morphologic neuronal differentiation similar to treatment with NGF. Oncogenic c-Ha-ras-induced neurite outgrowth is not stable with long-term culture, rather the cells revert to an undifferentiated morphology with altered cell cycle kinetics. 3. The stable neuronal phenotype induced by v-src and NGF is characterized by the functional expression of dihydropyridine-insensitive calcium currents.

Animals↗

Intracellular regulation of ion channels in cell membranes.

Cells communicate with their environment through receptor proteins on the cell membrane. Some ion channels are receptors, whereas others are linked to receptors through guanine nucleotide-binding proteins (G proteins). Ion channels control intracellular concentrations of ions such as calcium, and these concentrations control cell functions such as secretion and cell division. This review summarizes the current state of knowledge about the control of ion channels.

Cell Communication↗

Initial test of the benchmark chemical approach for predicting microbial transformation rates in aquatic environments.

Using 2,4-dichlorophenoxyacetic acid methyl ester (2,4-DME) as a benchmark chemical, we determined relative pseudo-first-order rate coefficients for the butoxyethyl ester of 2,4-dichlorophenoxyacetic acid (2,4-DBE), methyl parathion, and methyl-3-chlorobenzoate in a diversity of microbial samples, including water, sediment, biofilm, and floating microbial mats collected from a laboratory mesocosm as well as from streams, lakes, and wetlands in Georgia and Florida. The decreasing order of reactivity for relative microbial transformation rates was 2,4-DBE > 2,4-DME > methyl-3-chlorobenzoate > methyl parathion. Half-lives of the chemicals varied about 60-fold depending on the chemical and microbial sample. Relative rate coefficients, however, typically varied only about threefold for field-collected samples. Relative rate coefficients determined with samples from a laboratory mesocosm were consistently low compared with the field sample data. Overall, the data indicated that microbial transformation rates of a chemical can be satisfactorily inferred for a wide variety of microbial habitats-such as water, biofilm, or a sediment-on the basis of its transformation rate relative to that of an appropriate benchmark chemical by using a single type of microbial sample.

Journal Article↗

G-protein beta gamma-subunits activate the cardiac muscarinic K+-channel via phospholipase A2.

Muscarinic receptors of cardiac pacemaker and atrial cells are linked to a potassium channel (IK.ACh) by a pertussis toxin-sensitive GTP-binding protein. The dissociation of G-proteins leads to the generation of two potential transducing elements, alpha-GTP and beta gamma. IK.ACh is activated by G-protein alpha- and beta gamma-subunits applied to the intracellular surface of inside-out patches of membrane. beta gamma has been shown to activate the membrane-bound enzyme phospholipase A2 in retinal rods. Arachidonic acid, which is produced from the action of phospholipase A2 on phospholipids, is metabolized to compounds which may act as second messengers regulating ion channels in Aplysia. Muscarinic receptor activation leads to the generation of arachidonic acid in some cell lines. We therefore tested the hypothesis that beta gamma activates IK.ACh by stimulation of phospholipase A2. When patches were first incubated with antibody that blocks phospholipase A2 activity, or with the lipoxygenase inhibitor, nordihydroguaiaretic acid, beta gamma failed to activate IK.ACh. Arachidonic acid and several of its metabolites derived from the 5-lipoxygenase pathway, activated the channel. Blockade of the cyclooxygenase pathway did not inhibit arachidonic acid-induced channel activation. We conclude that the beta gamma-subunit of G-proteins activates IK.ACh by stimulating the production of lipoxygenase-derived second messengers.

Animals↗

Somatostatin activates an inwardly rectifying K+ channel in neonatal rat atrial cells.

Somatostatin, localized throughout the central and peripheral nervous systems, has been found in neurons of the vagal inhibitory pathway of the heart and has been shown to have negative inotopic effects in cardiac tissue. Using patch clamp techniques we show that somatostatin activates an inwardly rectifying K+ channel in rat atrial cells. Loss of somatostatin-induced K+ channel activity in excised inside-out patches is restored by the addition of GTP to the bath. Pertussis toxin pretreatment blocked GTP-dependent somatostatin activation of the inwardly rectifying K+ channel. This K+ channel has a conductance of 34 pS and a mean open time of approximately 1 ms. It is apparently the same K+ channel activated by muscarinic and adenosine receptors in atrial and cardiac pacemaker cells. Thus, atrial cells have at least three receptors which act via pertussis toxin-sensitive G proteins to activate the same class of K+ channels.

Animals↗

Human vitamin B-6 pools estimated through muscle biopsies.

Previous estimates of total vitamin B-6 pools in humans based on extrapolations from tracer studies yielded values of 107-190 mumol when the tracer was administered orally and 345-725 mumol when the tracer was administered intravenously. To obtain a more direct estimate of vitamin B-6 pools, muscle biopsies from five female and seven male adults were analyzed by cation-exchange chromatography. Total muscle mass was estimated from creatinine excretion and the assumption that muscle is 40% of the body weight. The total muscle vitamin B-6 pool was estimated to be 917 +/- 319 mumol in the females and 850 +/- 216 mumol in the males. Because muscle accounts for approximately 80% of the vitamin B-6 in the body, the total body pool of vitamin B-6 in adult humans is probably approximately 1000 mumol.

Adult↗

Prediction of substrate removal rates of attached microorganisms and of relative contributions of attached and suspended communities at field sites.

A mathematical model composed of a direct proportionality relationship between bulk water velocities and field-determined second-order microbial transformation rate coefficients, and the relative rate coefficient of a benchmark chemical, was developed for estimating the substrate removal rates of rapidly degraded chemicals by attached organisms in shallow (<1 m deep) aquatic ecosystems. Data from 31 field experiments involving the addition of 2,4-dichlorophenoxyacetic acid methyl ester (2,4-DME) in nine field areas were used to determine a field-derived second-order rate coefficient for microbial transformation of the ester. By using 2,4-DME as a benchmark chemical, the model was used to predict microbial transformation rates of the butoxyethyl ester of 2,4-dichlorophenoxyacetic acid (2,4-DBE) at five other field sites. The predicted half-lives of 2,4-DBE varied 1,500-fold and were within about a threefold range or less of the measured half-lives. Under conditions of mass transport limitation, the contributions of attached microorganisms relative to total microbial activities at various field sites were related to the ratio of water velocity, U, and depth, D, showing that historical definitions of ecosystems according to flow and depth characteristics are also valid for describing the process-related structure of ecosystems. An equation was developed for predicting the relative contributions of attached and suspended communities with values of U and D for lotic and lentic ecosystems. On the basis of this equation, attached microorganisms were expected to be insignificant in deep lentic ecosystems and suspended microorganisms were expected to be insignificant in shallow lotic systems for the same process carried out by both populations. Neglecting epiphytic microorganisms, both suspended and attached organisms were expected to be significant in wetlands.

Journal Article↗

Kinetics of mixed microbial assemblages enhance removal of highly dilute organic substrates.

Our experiments with selected organic substrates reveal that the rate-limiting process governing microbial degradation rates changes with substrate concentration, S, in such a manner that substrate removal is enhanced at lower values of S. This enhancement is the result of the dominance of very efficient systems for substrate removal at low substrate concentrations. The variability of dominant kinetic parameters over a range of S causes the kinetics of complex assemblages to be profoundly dissimilar to those of systems possessing a single set of kinetic parameters; these findings necessitate taking a new approach to predicting substrate removal rates over wide ranges of S.

Journal Article↗

The protein kinase C activator 1-oleoyl-2-acetylglycerol inhibits voltage-dependent Ca2+ current in the pituitary cell line AtT-20.

The role of protein kinase C in regulating Ca2+ channel activity was investigated using the whole-cell patch-clamp technique in the mouse pituitary tumor cell line AtT-20. The Ca2+ current was activated by depolarizing voltage steps from a holding potential of -80 mV. Extracellular application of the protein kinase C activator 1-oleoyl-2-acetylglycerol (OAG) reduced voltage-dependent Ca2+ current. This effect was reversible and dose dependent (10-100 microM). Pertussis toxin did not block the effect of OAG on Ca2+ current, suggesting that OAG does not affect Ca2+ channels via a pertussis toxin sensitive guanosine triphosphate binding protein. Na+-free solutions did not block the effect of OAG on Ca2+ channels, suggesting that this effect of OAG does not involve the Na+/H+ antiporter. The phorbol esters 12-deoxyphorbol-13-isobutyrate (10 microM) and phorbol-12,13-diacetate (100 microM) also reduced Ca2+ current. The results suggest that protein kinase C may be an inhibitory regulator of voltage-dependent Ca2+ channels.

Animals↗

Calcium currents and fura-2 signals in fluorescence-activated cell sorted lactotrophs and somatotrophs of rat anterior pituitary.

Optical and electrical recording techniques were applied to single primary pituitary cells to characterize the types of voltage-dependent calcium currents (ICa) and levels of intracellular calcium ([Ca2+]i). GH-containing somatotrophs and PRL-containing lactotrophs were isolated from adult female rats using fluorescence-activated cell-sorting techniques and were maintained in culture for 1-4 days. Whole cell patch-clamp recordings were made to analyze the ICa, and [Ca2+]i was measured with fura-2. Cell type was verified after each recording by indirect immunocytochemistry. GH and PRL cells could be divided into two groups: silent and spontaneously active. Silent cells had stable membrane potentials and stable levels of [Ca2+]i. Spontaneously active cells exhibited spontaneous action potentials and large fluctuations in [Ca2+]i. Two types of ICa were found: a low threshold, transient current which was insensitive to the dihydropyridine -Bay 5417 (the negative isomer of Bay K 8644), and a high threshold, sustained current which was enhanced by -Bay 5417. Both types of ICa were present in PRL and GH cells, but each cell type differed quantitatively in the proportion of each current type. While the GH cells had a more prominent, low threshold, transient ICa, the PRL cells had a more prominent, high threshold, sustained ICa. The enhancement of ICa by -Bay 5417 was greater in the PRL cells, which have a larger dihydropyridine-sensitive ICa. Parallel fura-2 measurements showed an increase in [Ca2+]i in response to 50 mM KCl and -Bay 5417 for both lactotrophs and somatotrophs.

Animals↗

Vasoactive intestinal peptide-containing nerves in Peyer's patches.

Vasoactive intestinal peptide (VIP) is a neuropeptide that is well represented in the gut tract. Previous work has demonstrated that murine T lymphocytes have high-affinity specific receptors for VIP and has implicated their interaction with VIP in the control of T-cell migration into Peyer's patches in vivo. It was postulated that this effect was mediated by interactions in the vicinity of the specialized endothelium of the postcapillary venules of Peyer's patches. We report the localization of VIP-like immunoreactivity in mouse Peyer's patches. Immunohistochemical staining was performed with heterologous antiserum against VIP using the avidin-biotin-peroxidase complex technique. VIP positivity was present near vessels of various sizes and in close proximity to small caliber vessels lined with specialized polygonal endothelial cells. These findings provide an anatomical basis for the concept that VIP may be available as a local neurophysiological signal during the migration of lymphocytes from the blood into Peyer's patches.

Animals↗

A guanine nucleotide-binding protein mediates the inhibition of voltage-dependent calcium current by somatostatin in a pituitary cell line.

Somatostatin reduces voltage-dependent Ca2+ current (ICa) and intracellular free Ca2+ concentration in the AtT-20/D16-16 pituitary cell line. We tested whether guanine nucleotide-binding proteins (G or N proteins) are involved in the signal transduction mechanism between the somatostatin receptor and voltage-dependent Ca2+ channels. Treatment of the cells with pertussis toxin, which selectively ADP ribosylates the GTP binding proteins Gi and Go and suppresses the ability of Gi to couple inhibitory receptors to adenylate cyclase, abolished the action of somatostatin on both ICa and intracellular free Ca2+. Intracellular application of the nonhydrolyzable guanine nucleotide analog guanosine 5'-[gamma-thio]triphosphate (GTP[gamma S]), which irreversibly activates G proteins, changed the somatostatin effect on ICa from a reversible to an irreversible inhibition. Intracellular GTP[gamma S] alone caused a very slowly developing inhibition of ICa. When ICa was inhibited by GTP[gamma S] (alone or with somatostatin), it failed to respond to subsequent applications of somatostatin. The effect of GTP[gamma S] on the inhibition of ICa by somatostatin was not altered by the intracellular application of cAMP and 3-isobutyl-1-methylxanthine. The results suggest that a GTP-binding protein is directly involved in the cAMP-independent receptor-mediated inhibition of voltage-dependent Ca2+ channels.

Adenylate Cyclase Toxin↗