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Biomedical subjects

G D Griffin

Publications and source records attributed to G D Griffin.

At least 19 recordsLinked to original sources
PubMed2006

Sexual behaviour induces the expression of activity-regulated cytoskeletal protein and modifies neuronal morphology in the female rat ventromedial hypothalamus.

Female sexual behaviour activates a distributed network within the brain, including the ventrolateral subdivision of the hypothalamic ventromedial nucleus (vlVMH), as demonstrated by behavioural studies performed in conjunction with the neuroanatomical analysis of immediate early gene (IEG) expression. However, it has been difficult to interpret mating-induced IEG expression because the precise function of many IEGs remains poorly defined. One possible function for genomic activation of the vlVMH during mating behaviour is to establish synaptic remodelling. The present experiments tested the hypothesis that sexual behaviour rapidly induces the expression of a structural protein associated with synaptic plasticity and ultimately causes morphological changes in the vlVMH. First, the expression of activity-regulated cytoskeletal protein (Arc), an IEG associated with neural plasticity, was assayed immunohistochemically in females after approximately 1 h of mating. The number of Arc-labelled neurones in the vlVMH was greater in mated females compared to unmated controls. Second, VMH neurones were biolistically labelled for morphological measurements, including soma size, dendrite number and length and dendritic spine density. Dendritic spine density in the vlVMH was significantly reduced 5 days after mating in experienced females compared to sexually naïve females. There were no differences between these groups in soma size, dendrite length or dendrite number. Collectively, these studies suggest that mating behaviour produces short-term changes in structural proteins and long-term, selective changes in dendrite morphology, which then may influence future behaviours and/or physiology.

Animals↗
PubMed2002

Nanosensor for in vivo measurement of the carcinogen benzo[a]pyrene in a single cell.

This work describes the fabrication and the application of an antibody-based fiber-optic nanosensor for in situ measurements of the carcinogen benzo[a]pyrene (BaP) in a single cell. This antibody-based spectroscopic nanosensor is miniaturized enabling the detection of fluorescent analytes in single cells. In addition to measuring fluorescent analytes in single cells, the nanosensor has the potential to be applied for both diagnostic and proteomics purposes. In this work, the human breast carcinoma cell line, MCF-7, was used as the model system to perform BaP measurements in single cells. A standard concentration curve for BaP was established and used to perform quantitative analyses of BaP in individual cells. From these analyses, it was estimated that the concentration of BaP in the individual cells investigated was approximately 3.61 x 10(-10) M. The results obtained demonstrate the application of antibody-based nanosensors for performing in situ measurements inside a single cell.

Antibodies↗
PubMed2001

Detection of E. coli using a microfluidics-based antibody biochip detection system.

This work demonstrates the detection of E. coli using a 2-dimensional photosensor array biochip which is efficiently equipped with a microfluidics sample/reagent delivery system for on-chip monitoring of bioassays. The biochip features a 4 x 4 array of independently operating photodiodes that are integrated along with amplifiers, discriminators and logic circuitry on a single platform. The microfluidics system includes a single 0.4 mL reaction chamber which houses a sampling platform that selectively captures detection probes from a sample through the use of immobilized bioreceptors. The independently operating photodiodes allow simultaneous monitoring of multiple samples. In this study the sampling platform is a cellulosic membrane that is exposed to E. coli organisms and subsequently analyzed using a sandwich immunoassay involving a Cy5-labeled antibody probe. The combined effectiveness of the integrated circuit (IC) biochip and the immunoassay is evaluated for assays performed both by conventional laboratory means followed by detection with the IC biochip, and through the use of the microfluidics system for on-chip detection. Highlights of the studies show that the biochip has a linear dynamic range of three orders of magnitude observed for conventional assays, and can detect 20 E. coli organisms. Selective detection of E. coli in a complex medium, milk diluent, is also reported for both off-chip and on-chip assays.

Animals↗
PubMed2000

Intracellular measurements in mammary carcinoma cells using fiber-optic nanosensors.

Submicrometer fiber-optic biosensors have been developed and used to measure toxic chemicals within single cells. Optical fibers that have been pulled to a distal-end diameter of less than 1 micrometer are coated with antibodies to selectively bind the species of interest. This paper describes the use of these fibers to selectively measure the concentration of benzo[a]pyrene tetrol (BPT), a metabolite of benzo[a]pyrene, within individual cells of two different cell lines, human mammary carcinoma cells and rat liver epithelial cells. The results from these measurements have been used to determine the sensitivity, reproducibility, and usefulness of these nanosensors. The detection limit of these biosensors has been determined to be 0.64 +/- 0.17 x 10(-11) M for BPT.

Animals↗
PubMed2000

Power frequency magnetic field exposure and gap junctional communication in Clone 9 cells.

Exposure to a power-frequency magnetic field has been reported to produce a statistically significant inhibition of gap junctional communication (GJC) in Clone 9 cells that have been pre-stressed by treatment with low concentrations of chloral hydrate (CH) [C.F. Blackman, J.P. Blanchard, S.G. Benane, D.E. House, J.A. Elder, Double blind test of magnetic field effects on neurite outgrowth, Bioelectromagnetics, 19 (1998) 204-209]. This observation might provide mechanistic insight into the possible role of electromagnetic fields (EMFs) in the carcinogenic process, since cancer cells frequently show decreased or absent GJC, and tumor promoting chemicals have been observed to inhibit GJC. Magnetic field exposure conditions were 45 Hz, 23.8 microT rms + parallel DC 36.6 microT, for 30 min of exposure. The responses of Clone 9 cells to the GJC-inhibiting effects of the tumor promoter 12-O-tetradecanoylphorbol 13-acetate and the chemical CH were evaluated and compared to reported results [S.G. Benane, C.F. Blackman, D.E. House, Effects of perchloroethylene and its metabolites on intercellular communication in Clone 9 rat liver cells, J. Toxicol. Environ. Health, 48 (1996) 427-437]. Before magnetic field exposure, cells were exposed for 24 h to either 3 (nine experiments) or 5 mM (11 experiments) CH to produce GJC of 67% or 50%, respectively, relative to unexposed controls. GJC was assessed microscopically using the scrape-loading technique and a blinded protocol. No statistically significant effect was observed due to magnetic field exposure with either CH concentration.

Animals↗
PubMed2000

Antibody-based nanoprobe for measurement of a fluorescent analyte in a single cell.

We report here the application of an antibody-based nanoprobe for in situ measurements of a single cell. The nanoprobe employs antibody-based receptors targeted to a fluorescent analyte, benzopyrene tetrol (BPT), a metabolite of the carcinogen benzo[a]pyrene (BaP) and of the BaP-DNA adduct. Detection of BPT is of great biomedical interest, since this species can serve as a biomarker for monitoring DNA damage due to BaP exposure and for possible precancer diagnosis. The measurements were performed on the rat liver epithelial clone 9 cell line, which was used as the model cell system. Before making measurements, the cells were treated with BPT. Nanoprobes were inserted into individual cells, incubated 5 min to allow antigen-antibody binding, and then removed for fluorescence detection. We determined a concentration of 9.6+/-0.2x10(-11) M for BPT in the individual cells investigated. The results demonstrate the possibility of in situ measurements inside a single cell using an antibody-based nanoprobe.

Animals↗
PubMed2000

Cellular communication in clone 9 cells exposed to magnetic fields.

Magnetic-field exposure (45 Hz B(a.c.) over a flux density range of 7.7 to 49.9 microT r.m.s. with parallel B(d.c.) of 36.6 microT) has been reported by Blackman and coworkers to inhibit gap junction intercellular communication in Clone 9 cells treated with chloral hydrate for 24 h prior to field exposure in accord with predictions of the ion parametric resonance model. The study reported here is an attempt to reproduce this effect. Baseline experiments showed that growth in culture and state of confluence at time of addition of chloral hydrate were comparable in both laboratories. PMA inhibited cell-cell communication in a dose-dependent manner, similar to the results of Blackman and coworkers, whereas cells in the present study were somewhat more sensitive to chloral hydrate than reported by Blackman and coworkers. A total of 38 exposure experiments were undertaken using a 45 Hz magnetic field with a flux density of 23.8 microT r.m.s., in parallel with a 36.6-microT static magnetic field for 40 to 45 min, after pretreatment with 2.5 mM chloral hydrate for 24 h. In 14 unblinded experiments, a small but statistically significant effect of magnetic-field exposure was observed, but due to the subjective nature of the assay, it was deemed essential to carry out blinded experiments. The remaining 24 experiments were blinded. In 15 blinded experiments, cells purchased from the American Type Culture Collection and grown only in this laboratory were used, while in 9 experiments, the cells had originally been grown in Blackman's laboratory and were subsequently sent to this laboratory. There was no statistically significant effect of magnetic-field exposure on gap junction intercellular communication in these blinded experiments using either cell line.

Animals↗
PubMed1999

The sources, fate, and toxicity of chemical warfare agent degradation products.

We include in this review an assessment of the formation, environmental fate, and mammalian and ecotoxicity of CW agent degradation products relevant to environmental and occupational health. These parent CW agents include several vesicants: sulfur mustards [undistilled sulfur mustard (H), sulfur mustard (HD), and an HD/agent T mixture (HT)]; nitrogen mustards [ethylbis(2-chloroethyl)amine (HN1), methylbis(2-chloroethyl)amine (HN2), tris(2-chloroethyl)amine (HN3)], and Lewisite; four nerve agents (O-ethyl S-[2-(diisopropylamino)ethyl] methylphosphonothioate (VX), tabun (GA), sarin (GB), and soman (GD)); and the blood agent cyanogen chloride. The degradation processes considered here include hydrolysis, microbial degradation, oxidation, and photolysis. We also briefly address decontamination but not combustion processes. Because CW agents are generally not considered very persistent, certain degradation products of significant persistence, even those that are not particularly toxic, may indicate previous CW agent presence or that degradation has occurred. Of those products for which there are data on both environmental fate and toxicity, only a few are both environmentally persistent and highly toxic. Major degradation products estimated to be of significant persistence (weeks to years) include thiodiglycol for HD; Lewisite oxide for Lewisite; and ethyl methyl phosphonic acid, methyl phosphonic acid, and possibly S-(2-diisopropylaminoethyl) methylphosphonothioic acid (EA 2192) for VX. Methyl phosphonic acid is also the ultimate hydrolysis product of both GB and GD. The GB product, isopropyl methylphosphonic acid, and a closely related contaminant of GB, diisopropyl methylphosphonate, are also persistent. Of all of these compounds, only Lewisite oxide and EA 2192 possess high mammalian toxicity. Unlike other CW agents, sulfur mustard agents (e.g., HD) are somewhat persistent; therefore, sites or conditions involving potential HD contamination should include an evaluation of both the agent and thiodiglycol.

Animals↗
PubMed1997

Low-cost system for real-time monitoring of luciferase gene expression.

In some mammalian cells transfected with luciferase reporter genes, the luciferase/luciferin reaction in a cell monolayer produces a very small light flux. While the low light levels are often measurable with single-photon counting cameras, these devices are expensive and may require long averaging times to acquire an image. We describe an approach for real-time monitoring of light produced by luciferase gene expression in intact, cultured cells using readily available and relatively inexpensive components. The system uses a single-photon counting photomultiplier tube with built-in high voltage supply and photon counting circuitry to rapidly measure average light output from growing cells in a 35 mm culture dish. The fast, accurate and highly sensitive response of the system makes it useful for studying the dynamics of gene expression over time periods ranging from minutes to days.

Antineoplastic Agents, Hormonal↗
PubMed1997

Studying electric field effects on embryonic myocytes.

Several problems arise when electrophysiological measurements are attempted on cells exposed to an electric field. In addition to field distortion produced by the reference electrode, membrane potential measurements by conventional microelectrode or patch-clamp techniques suffer serious interference from the applied field. We describe here a novel method for measurement of cardiac myocyte response to an alternating electric field that avoids these problems by sensing the mechanical activity of the cells rather than their electrical activity. A miniature electromechanical force transducer is used for this purpose. A glass pipet is attached to the force transducer, and only this pipet makes actual contact with the cell preparation. The resistive elements of the transducer are arranged as two legs of a Wheatstone bridge. Contractile activity of the cells produces small displacements of the micropipet and a resulting change in the transducer resistances. The Wheatstone bridge output is a current signal that is detected and converted to a voltage signal by a picoammeter before amplification and recording for later analysis. The technique may find applications in a variety of experimental studies of contractile tissues.

Action Potentials↗
PubMed1994

A flow cytometric method for phenotyping recipient red cells following transfusion.

BACKGROUND: Reticulocyte phenotyping is used for transfused patients, who have red cell antibodies, to match blood for subsequent transfusion. Current methods are labor-intensive and require a significant amount of sample. STUDY DESIGN AND METHODS: A simple dual-color flow cytometry method developed for antigen typing of reticulocytes in mixed red cell populations is reported. Antigens were labeled by an indirect immunofluorescence technique using undiluted reagent sera as the primary label, biotinylated goat anti-human IgG as the secondary label, and avidin-phycoerythrin as the fluorescent stain. Reticulocytes were labeled with a thiazole orange fluorescent stain. Reticulocyte identification and antigen typing were performed on 319 samples to establish the validity of the procedure. Mixed red cells were prepared in all possible c antigen combinations to simulate transfusion concentrations of 25, 50, and 75 percent. RESULTS: The anti-c flow cytometry profiles readily distinguished between antigen-positive and antigen-negative populations and allowed the detection of reticulocytes at all simulated transfusion concentrations. Similar results were obtained in experiments using C, K, s, Fya, Fyb, Jka, or Jkb sera against equal volumes of antigen-positive and -negative cells. Anti-S gave inconsistent results. The in vitro results were confirmed in 19 transfused patients who had received red cells antigenically different from their own as well as cells from 1 chimera blood donor. CONCLUSION: This method provides a simpler, safer, less labor-intensive, and less subjective technique requiring far less sample volume than current methods for antigen typing of reticulocytes in mixed red cell samples from recently transfused patients.

Antigens, Surface↗
PubMed1992

Toxicity of vesicant agents scheduled for destruction by the Chemical Stockpile Disposal Program.

The vesicant agents of the unitary chemical munitions stockpile include various formulations of sulfur mustard [bis-(2-chloroethyl) sulfide; agents H, HD, and HT] and small quantities of the organic arsenical Lewisite [dichloro(2-chlorovinyl) arsine; agent L]. These agents can be dispersed in liquid, aerosol, or vapor form and are capable of producing severe chemical burns upon direct contact with tissue. Moist tissues such as the eyes, respiratory tract, and axillary areas are particularly affected. Available data summarizing acute dose response in humans and laboratory animals are summarized. Vesicant agents are also capable of generating delayed effects such as chronic bronchitis, carcinogenesis, or keratitis/keratopathy of the eye under appropriate conditions of exposure and dose. These effects may not become manifest until years following exposure. Risk analysis derived from carcinogenesis data indicates that sulfur mustard possesses a carcinogenic potency similar to that of benzo[a]pyrene. Because mustard agents are alkylating compounds, they destroy individual cells by reaction with cellular proteins, enzymes, RNA, and DNA. Once begun, tissue reaction is irreversible. Mustard agents are mutagenic; data for cellular and laboratory animal assays are presented. Reproductive effects have not been demonstrated in the offspring of laboratory rats. Acute Lewisite exposure has been implicated in cases of Bowen's disease, an intraepidermal squamous cell carcinoma. Lewisite is not known to generate reproductive or teratogenic effects.

Animals↗
PubMed1990

Treating exposure to chemical warfare agents: implications for health care providers and community emergency planning.

Current treatment protocols for exposure to nerve and vesicant agents found in the U.S. stockpile of unitary chemical weapons are summarized, and the toxicities of available antidotes are evaluated. The status of the most promising of the new nerve agent antidotes is reviewed. In the U.S. atropine and pralidoxime compose the only approved antidote regimen for organophosphate nerve agent poisoning. Diazepam may also be used if necessary to control convulsions. To avoid death, administration must occur within minutes of substantial exposure together with immediate decontamination. Continuous observation and repeated administration of antidotes are necessary as symptoms warrant. Available antidotes do not necessarily prevent respiratory failure or incapacitation. The toxicity of the antidotes themselves and the individualized nature of medical care preclude recommending that autoinjectors be distributed to the general public. In addition, precautionary administration of protective drugs to the general population would not be feasible or desirable. No antidote exists for poisoning by the vesicant sulfur mustard (H, HD, HT); effective intervention can only be accomplished by rapid decontamination followed by palliative treatment of symptoms. British anti-Lewisite (BAL) (2,3-dimercapto-1-propanolol) is the antidote of choice for treatment of exposure to Lewisite, another potent vesicant. Experimental water-soluble BAL analogues have been developed that are less toxic than BAL. Treatment protocols for each antidote are summarized in tabular form for use by health care providers.

Antidotes↗
PubMed1989

Sulfur mustard as a carcinogen: application of relative potency analysis to the chemical warfare agents H, HD, and HT.

A relative potency method for assessing potential human health effects from exposures to relatively untested chemicals is presented and documented. The need for such a method in evaluating the carcinogenic potential of the chemical warfare agent sulfur mustard (agent HD) from a limited data base is specifically addressed. The best-estimate potency factor for sulfur mustard relative to benzo[a]pyrene is 1.3, with an interquartile range of 0.6 to 2.9. The method is applied to (1) the estimated fence-boundary air concentrations of mustard during operation of a proposed agent incinerator at Aberdeen Proving Ground (APG), Maryland, and (2) the current approved general population exposure level of 1 X 10(-4) mg HD/m3 and the occupational exposure level of 3 X 10(-3) mg HD/m3. Maximum estimates of excess lifetime cancer risk for individuals at sites along the APG boundary range between 3 X 10(-8) and 1 X 10(-7). Lifetime cancer risk estimates less than or equal to 10(-6) are not now regulated by the U.S. Environmental Protection Agency or the Food and Drug Administration. Maximum estimates of excess lifetime cancer risk assuming daily exposure to the approved standards during the proposed 5 years of incinerator operation are on the order of 10(-5) for the general public and 10(-4) for the worker population. These values are considered upper limit estimates.

Animals↗
PubMed1986

Induction of mixed-function oxidase activity in mouse lymphoid tissues by polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbon (PAH) exposure can cause mixed-function oxidase (MFO) enzyme induction in certain tissues of various organisms. Measurement of such induction might serve as a useful bioindicator of human exposure to PAHs, provided readily obtainable human tissues can be utilized for such measurements. We have investigated the MFO activity in various lymphoid tissues of the C3H mouse as a model system and have studied the effect of systemic PAH treatment on such enzyme activity. An MFO enzyme assay was used to measure the activity of 7-ethoxyresorufin deethylase, an enzyme activity that may be specific for the cytochrome P-448 subset of MFO enzymes (those enzymes that are induced in cells or tissues following PAH administration). Intraperitoneal injection of mice with 180 mg/kg (4.6 mg) benzo[a]pyrene (BaP) or 160 mg/kg (4.0 mg) 3-methylcholanthrene (MC) produced a significant induction in MFO activity in mouse spleen S9 fractions 48 h after the injection. Induction ratios (induced activity/control activity) between 4 and 5 were seen with BaP; MC produced induction ratios of 2.5-3.0. Enzyme activity was not induced in the spleen within 16 h following BaP or MC administration. Other experiments indicated that MFO activity could be induced in thymus cells 48 h after either BaP or MC treatment. Treatment with BaP or MC did produce significant enzyme induction in the liver and lung tissues from the animals both 16 and 48 h after chemical treatment.

Animals↗
PubMed1986

Inhibition of murine interferon production following in vivo administration of benzo[a]pyrene.

To investigate whether alteration in interferon (IFN) production might serve as a biomarker for certain toxic chemical exposures, an in vivo mouse model system was studied. Female C3H mice were injected intraperitoneally with varying doses of benzo[a]pyrene (BP), and at various times subsequent to this treatment, serum IFN levels, following Sendai virus induction, were determined by a cytopathic effect inhibition assay. Doses of 4.6 mg/mouse (180 mg/kg body weight) caused a significant depression in IFN production at 12, 48, and 120 h after BP administration. Doses of 0.46 mg also produced significant decreases at 48 h following exposure. At 48 h post-BP treatment, the reduction in serum IFN titers in treated animals relative to controls was: 62% for the 0.46-mg dose, and 94% for the 4.6-mg dose. These results indicate that systemic administration of BP can significantly depress the whole-animal IFN response to viral stimulation, and that such depression can persist for a rather extended period at certain dose levels.

Analysis of Variance↗