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PubMed1991

Computer-aided design and evaluation of novel catheters for conductive interstitial hyperthermia.

Conductive interstitial heating is a modality in which heating elements are implanted directly into the treated tissue. One implementation of such therapy employs electrically heated catheters that are implanted in staggered, parallel rows. To explore strategies for maximising the uniformity of tissue temperature distributions achieved with heated catheters, a two-dimensional computer model with cylindrical co-ordinates was used to evaluate radially and longitudinally the temperature distributions produced by a typical interior catheter surrounded by other similar catheters. Insights from the computer model led to new designs for catheters containing multiple heating elements that produced more uniform thermal distributions, eliminating previous 'cold spots' within the treatment volume located near the ends of the catheter. The new catheter designs also include compartments for the optional placement of radioactive seeds for simultaneous thermoradiotherapy.

Biomedical Engineering↗
PubMed2001

The grp78 promoter of Neurospora crassa: constitutive, stress and differentiation-dependent protein-binding patterns.

In order to characterize the transcriptional regulation of the grp78 (glucose-regulated protein 78 kDa) gene of Neurospora crassa, a 1.65-kb genomic fragment upstream of the protein-coding region was sequenced and analysed. A single transcription start point was mapped 160 nt upstream of the first codon. Several distinct protein-DNA binding sites were identified in the promoter region by non-radioactive scanning electrophoretic mobility shift analysis during growth, stress treatment and differentiation of conidia. A protein DNA binding complex induced by tunicamycin was linked to a promoter motif similar to the unfolded protein response element consensus of yeast. Another binding complex in differentiating aerial hyphae was found, which differs from the known cis-elements involved in conidiation-dependent gene expression.

Base Sequence↗
PubMed2001

The availability of plutonium and americium in Irish Sea sediments for re-dissolution.

The availability of plutonium and americium, for re-dissolution from offshore sediments into Irish Sea water, has been examined. Sediments collected from the mud-patch near the Cumbrian coast were characterized in terms of spatial location, particle size, partitioning of radionuclides with respect to physico-chemical bonds and availability of actinides for release into seawater. Sequential extraction investigations revealed that plutonium was predominantly associated with strongly bound sesquioxide and organic complex fractions. Americium was associated mainly with the organic complex fraction, but a significant fraction was in carbonate form. Sediment/water re-dissolution experiments with and without stirring were compared to simulate the effect of disturbing bed sediment. After 1 week, neither set of re-dissolution data provided significant trends between dissolved activity and time. Stirred systems appeared to release 2.5 times more plutonium and americium into seawater than unstirred systems. Measured 239,240Pu and 241Am distribution coefficients (Kd values) were both typically approximately 10(5) l kg(-1). 241Am Kd values are an order of magnitude lower than previously reported for the north-eastern Irish Sea, but similar to western Irish Sea values. Overall, the fractions of plutonium and americium available for re-dissolution from bed sediment are very low at < 0.1%, with proportionally more plutonium being released than americium. These findings lend further support for the extrapolation of laboratory-derived information to environmental conditions.

Actinoid Series Elements↗
PubMed1993

Continuous monitoring of cerebral perfusion by thermal clearance.

Currently, no commercially available system exists to continuously monitor the effective tissue perfusion within the parenchyma of the brain. While several methods exist for accurately measuring cerebral perfusion; among them: 133xeonon clearance, hydrogen clearance and radiolabeled microsphere injection; none of these methods provides continuous monitoring. The Cook Incorporated VH8500 Volumetric Hyperthermia Treatment System (Bloomington, IN, USA) was initially developed to treat brain tumours by maintaining constant, moderate hyperthermia within a defined tissue volume over an extended duration. The system continuously adjusts the power applied to heating elements in order to maintain a constant temperature within the treatment volume. Because tissue perfusion is a primary factor responsible for removing heat from tissue, monitoring the amount of power applied to the heating elements allows one to continuously estimate tissue perfusion in the vicinity of the heating elements. In the current study, regional blood flow in the vicinity of heater/sensor catheters implanted in the brain parenchyma of three dogs was estimated by the VH8500 tissue perfusion algorithm and directly measured with radioactive labeled microspheres. The accuracy of the perfusion estimate (Thermal Perfusion Index) was evaluated by comparing these values. A range of blood flow was achieved in each animal by infusing nitroprusside. It was found that with the perfusion estimation algorithm of the Cook Incorporated VH8500 Volumetric Hyperthermia Treatment System as it is currently implemented, the Thermal Perfusion Index tended to underestimate regional perfusion as measured with radioactive microspheres, but the relationship was nearly linear. Thus, the system currently tracks changes in regional blood flow.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1981

Incorporation of axonally transported glycoproteins into axolemma during nerve regeneration.

The insertion of axonally transported fucosyl glycoproteins into the axolemma of regenerating nerve sprouts was examined in rat sciatic motor axons at intervals after nerve crush. [(3)H]Fucose was injected into the lumbar ventral horns and the nerves were removed at intervals between 1 and 14 d after labeling. To follow the fate of the "pulse- labeled" glycoproteins, we examined the nerves by correlative radiometric and EM radioautographic approaches. The results showed, first, that rapidly transported [(3)H]fucosyl glycoproteins were inserted into the axolemma of regenerating sprouts as well as parent axons. At 1 d after delivery, in addition to the substantial mobile fraction of radioactivity still undergoing bidirectional transport within the axon, a fraction of label was already associated with the axolemma. Insertion of labeled glycoproteins into the sprout axolemma appeared to occur all along the length of the regenerating sprouts, not just in sprout terminals. Once inserted, labeled glycoproteins did not undergo extensive redistribution, nor did they appear in sprout regions that formed (as a result of continued outgrowth) after their insertion. The amount of radioactivity in the regenerating nerves decreased with time, in part as a result of removal of transported label by retrograde transport. By 7-14 d after labeling, radioautography showed that almost all the remaining radioactivity was associated with axolemma. The regenerating sprouts retained increased amounts of labeled glycoproteins; 7 or 14 d after labeling, the regenerating sprouts had over twice as much of radioactivity as comparable lengths of control nerves or parent axons. One role of fast axonal transport in nerve regeneration is the contribution to the regenerating sprout of glycoproteins inserted into the axolemma; these membrane elements are added both during longitudinal outgrowth and during lateral growth and maturation of the sprout.

Animals↗
PubMed1989

A microcomputer algorithm for solving first-order compartmental models involving recycling.

A general algorithm for solving first-order compartmental models including recycling systems has been developed and its implementation on a microcomputer is described. Matrix algebra is used to obtain for any compartmental model an analytical solution, which is expressed as the exponential of a matrix of rate constants. A special technique is used in the algorithm to enable this exponential to be evaluated with a rapidly converging series. Truncation errors incurred in this process are estimated automatically. Thus, in an extreme case, where these errors may be significant, the appropriate action can be taken. Given a particular model, the user enters the model parameters into a rate matrix according to a simple rule. The algorithm then uses this matrix to solve the model, and thus no specialized mathematical knowledge is needed. The algorithm is given in a short BASIC program (60 lines) listed in an appendix. No additional software is required. By running this program on a standard microcomputer, the user can solve models of any complexity: those up to 15 compartments in seconds and those up to 30 compartments within a minute. The algorithm is thus ideally suited to solve kinetic models describing the transport of radionuclides in the environment or the translocation of elements in biological systems such as the metabolic models recommended by the International Commission on Radiological Protection (ICRP). Given the initial amount of material in each compartment at time t = 0, together with its radioactive decay constant, the algorithm gives both the amount in each compartment at any future time t and the number of disintegrations that will have occurred in each compartment up to time t. The computer program, shown in an appendix, could easily be used to calculate disintegrations over any time interval of interest, or to predict the quantities or fractions of an intake expected to be present in any in vivo or excretion compartments of interest. Thus, the algorithm can be useful in both the design and conduct of bioassay and internal dose assessment procedures.

Algorithms↗
PubMed1972

Studies of the interaction of 5-hydroxytryptamine and the perivascular innervation of the guinea-pig caecum.

1. The action and interaction of noradrenaline (NA), 5-hydroxytryptamine (5-HT) and the sympathetic innervation was studied in the isolated taenia of the guinea-pig caecum.2. Addition of 5-HT led to a contraction of the taenia while addition of NA or perivascular nerve stimulation resulted in relaxation. Responses to 5-HT or perivascular nerve stimulation were abolished by tetrodotoxin. Tetrodotoxin did not affect responses to applied NA. Hexamethonium and hyoscine converted the 5-HT response to a relaxation and augmented the relaxation which followed low frequency perivascular nerve stimulation. Hexamethonium and hyoscine did not affect the dose-response relationship for NA.3. Fatigue of mechanical responses of the taenia to perivascular nerve stimulation was accelerated when nerves were stimulated in the presence of 5-HT or alpha-methyl-p-tyrosine (alpha-MPT). These two agents were additive in this action.4. Reserpine, 6-hydroxydopamine and alpha-MPT all reduced the NA content of the taenia. However, only after 6-hydroxydopamine could adrenergic activity be related to NA content.5. Segments of taenia were incubated with either tritiated NA or 5-HT. An increased rate of release of radioactivity followed perivascular nerve stimulation after incubation with either substance. This release did not occur when tissue was taken from animals given reserpine or 6-hydroxydopamine.6. It is concluded that 5-HT activates neural elements exclusively while NA has a direct effect on smooth muscle. 5-HT can apparently be taken up by adrenergic axons, and appears to enter the releasable neurotransmitter pool. Since none of the actions characteristic of 5-HT are seen when it is released by adrenergic axons as a false neurotransmitter, the released amine probably fails to reach neuronal receptors for 5-HT.

Action Potentials↗
PubMed2000

Tissue distribution and pharmacological potential of SM-16896, a novel oestrogen-bisphosphonate hybrid compound.

Postmenopausal osteoporosis is caused mainly by a deficiency of oestrogen with rapid bone loss. To target oestrogen to the bone effectively, we have synthesized and evaluated the effects of a novel hybrid compound of oestrogen and bisphosphonate, SM-16896. The tissue distribution pattern and pharmacological potential are reported. Although the affinity for calf uterine oestrogen receptor was very low (IC50: 73.3 microM; 1/25000 of that of 17beta-oestradiol (2.84 nM)), SM-16896 showed oestrogenic activity. SM-16896 (1 microM) induced a 4.5-fold transcriptional activity in rat osteosarcoma UMR-106 cells compared with vehicle-treated control, when we used the expression vector for human oestrogen receptor and a CAT reporter plasmid containing an oestrogen-responsive element. The distribution of SM-16896 after a subcutaneous administration to 7-week-old female rats was examined by radioluminography using 3H-labelled SM-16896. At 30 min after the administration, significant radioactivity was detected in the bone. At 24 h after administration, a high level of radioactivity was detected in the bone, but in the uterus it was only at a background level. Daily subcutaneous administration of 0.5 mgkg(-1) SM-16896 for 12 weeks (five times per week) to 13-week-old ovariectomized rats suppressed the ovariectomized-induced reduction in bone mineral density. A bone mineral density ratio of 120% was maintained compared with sham-operated rats, whereas a relatively low suppression of uterine weight was observed (about 50% loss compared with sham-operated rats). In the same experiment, the implantation of a 17beta-oestradiol time-release pellet (0.25 mg/pellet/90 days) almost completely suppressed the reduction of both the bone mineral density and uterine tissue weight. It is likely that the effect of SM-16896 on bone was due to its oestrogenic activity, since 1.0 mgkg(-1) SM-18108, the bisphosphonate moiety of this compound, had no effect on bone in 7-week-old ovariectomized rats. The results suggest that SM-16896, a bisphosphonate-conjugated oestrogen, showed a preference profile in the uterus and bone due to its characteristic distribution pattern compared with the natural oestrogen analogue 17beta-oestradiol. Thus, bisphosphonate-conjugated oestrogens have the potential to improve patient compliance in oestrogen therapy by minimizing adverse effects and reducing the frequency of medication.

Animals↗
PubMed1980

Passage of extravascular tracers into canine jugular veins and carotid arteries.

Biologically active substances arising in the interstitial space (cell-fiber matrix) of a medium-sized blood vessel or in surrounding tissue might reach the lumen by diffusion. Substances so delivered to the vessel wall-blood interface would be situated to effectively influence the endothelium and to initiate deposition of blood elements on the luminal surface of vessels, thereby contributing to thrombosis and atherosclerosis. This study showed that 125I, 125I-albumin and 125I-fibrinogen passed across the walls of segments of canine jugular veins and carotid arteries that were maintained under 20 cm H2O presure while being suspended in solutions containig the radioactive substances. The ratio of 125I to 125I-protein (albumin, fibrinogen) was predictably greatly increased by diffusion across the vessel wall. Frozen sections cut parallel with the luminal surface of flattened segments of vessels showed a gradient of radioactivity from adventitial to luminal surface of the vessels. Part of the fibrogen (but no albumin) that had reached the inside of the vessel had been broken down into fragments. These observations show that ions and proteins originating in perivascular fluid transverse the wall of medium-sized veins and arteries, presumably by diffusion across complex water-filled channels. Thus biologically active substances arising in perivascular tissue or in the vessel wall itself can be expected to reach the luminal surface of the vessel wall where they can influence endothelium and/or initiate deposition of blood elements on the vessel wall. Furthermore, the action of proteolytic enzymes on protein molecules such as zymogens might give rise to active substances not previously present.

Animals↗
PubMed1993

Use of the insertion element IS6110 for DNA fingerprinting of Mycobacterium tuberculosis isolates presenting various profiles of drug susceptibility.

IS6100 is an insertion sequence of the IS3 family and it is present in multiple copies in the chromosome of Mycobacterium tuberculosis. Four to 15 copies are present in various strains of M. tuberculosis. In this study, the value of IS6110 as an epidemiological marker of tuberculosis was examined. Unrelated clinical strains from Greek patients presented, in restriction fragment length polymorphism analysis, a high degree of polymorphism, whereas patterns of related clinical strains from familial outbreaks were identical. Since RFLP analysis with acetylaminofluorene labeled IS6110 as the probe gave satisfactory results, it is suggested that this non-radioactive probe can be used in hospitals and health centres for the epidemiological survey of M. tuberculosis infections.

2-Acetylaminofluorene↗
PubMed2006

(137)Cs and (90)Sr uptake by sunflower cultivated under hydroponic conditions.

The (90)Sr and (137)Cs uptake by the plant Helianthus annuus L. was studied during cultivation in a hydroponic medium. The accumulation of radioactivity in plants was measured after 2, 4, 8, 16 and 32 days of cultivation. About 12% of (137)Cs and 20% of (90)Sr accumulated during the experiments. We did not find any differences between the uptake of radioactive and stable caesium and strontium isotopes. Radioactivity distribution within the plant was determined by autoradiography. (137)Cs was present mainly in nodal segments, leaf veins and young leaves. High activity of (90)Sr was localized in leaf veins, stem, central root and stomata. The influence of stable elements or analogues on the transfer behaviour was investigated. The percentage of non-active caesium and strontium concentration in plants decreased with the increasing initial concentration of Cs or Sr in the medium. The percentage of (90)Sr activity in plants decreased with increasing initial activity of the nuclide in the medium, but the activity of (137)Cs in plants increased. The influence of K(+) and NH(4)(+) on the uptake of (137)Cs and the influence of Ca(2+) on the uptake of (90)Sr was tested. The highest accumulation of (137)Cs (24-27% of the initial activity of (137)Cs) was found in the presence of 10 mM potassium and 12 mM ammonium ions. Accumulation of about 22% of initial activity of (90)Sr was determined in plants grown on the medium with 8 mM calcium ions.

Cesium Radioisotopes↗
PubMed1975

Studies on intracellular transport in the rat exocrine pancreas. I. Inhibition by aromatic amino acids in vitro.

In vitro incubation of rat pancreatic lobules in the presence of 10 mM concentrations of 2 natural (phenylalanine, tryptophane) and 2 modified aromatic amino acids (p-fluorophenylalanine, p-chlorophenylalanine) induces paracrystal formation in the cisternal space of the rough endoplasmic reticulum and in the acinar lumen. Aggregation of secretory material in transitional elements of the rough endoplasmic reticulum suggests tubular connection to the Golgi complex. Paracrystal formation is correlated with a disturbance of the three major phases in the secretory process of the exocrine cell. Incorporation of radioactive amino acids into proteins is inhibited by 10 mM concentrations of phenylalanine and tryptophane by 20 and 50% respectively and by p-chlorophenylalanine at 1 and 10 mM concentrations by 50 and 75%. The inhibition of protein synthesis is not due to a reduced intracellular concentration of radioactive precursor amino acids. Intracellular transport of newly synthesized proteins as studied by a radioassay for zymogen discharge and by cell fractionation is similarly inhibited by phenylalanine, tryptophane and p-chlorophenylalanine at 10 mM concentrarions (20, 30, and 40% respectively). Discharge of zymogens as measured by the secretion of amylase stimulated with 5 X 10(-6) M carbamylcholine is reduced by 20% if 10 mM concentrations of phenylalanine, tryptophane or p-chlorophenylalanine are present in the medium. Paracrystals were isolated by differential centrifugation and their protein content compared with isolated zymogen granules. On sodium dodecylsulfate gel electrophoresis paracrystalline proteins show the same electrophoretic pattern as the content of zymogen granules.

Amylases↗
PubMed1967

Intracellular transport of secretory proteins in the pancreatic exocrine cell. I. Role of the peripheral elements of the Golgi complex.

It has been established by electron microscopic radioautography of guinea pig pancreatic exocrine cells (Caro and Palade, 1964) that secretory proteins are transported from the elements of the rough-surfaced endoplasmic reticulum (ER) to condensing vacuoles of the Golgi complex possibly via small vesicles located in the periphery of the complex. To define more clearly the role of these vesicles in the intracellular transport of secretory proteins, we have investigated the secretory cycle of the guinea pig pancreas by cell fractionation procedures applied to pancreatic slices incubated in vitro. Such slices remain viable for 3 hr and incur minimal structural damage in this time. Their secretory proteins can be labeled with radioactive amino acids in short, well defined pulses which, followed by cell fractionation, makes possible a kinetic analysis of transport. To determine the kinetics of transport, we pulse-labeled sets of slices for 3 min with leucine-(14)C and incubated them for further +7, +17, and +57 min in chase medium. At each time, smooth microsomes ( = peripheral elements of the Golgi complex) and rough microsomes ( = elements of the rough ER) were isolated from the slices by density gradient centrifugation of the total microsomal fraction. Labeled proteins appeared initially (end of pulse) in the rough microsomes and were subsequently transferred during incubation in chase medium to the smooth microsomes, reaching a maximal concentration in this fraction after +7 min chase incubation. Later, labeled proteins left the smooth microsomes to appear in the zymogen granule fraction. These data provide direct evidence that secretory proteins are transported from the cisternae of the rough ER to condensing vacuoles via the small vesicles of the Golgi complex.

Animals↗
PubMed1994

An inhibitory factor in rat thymus which interferes with binding of cytosol Ah receptor to xenobiotic responsive element.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is known to induce aryl hydrocarbon hydroxylase in several tissues in the rat. This effect of TCDD is mediated by the Ah receptor. However, TCDD does not significantly enhance aryl hydrocarbon hydroxylase activity in the rat thymus. Analysis of 3H-TCDD bound Ah receptor on sucrose gradients revealed the presence of radioactivity sedimenting at 8-9S and 5-6S in the cytosol and nuclei of thymus respectively. Incubation of rat thymic cytosol with TCDD increased protein kinase C dependent phosphorylation, which reached saturation levels at high concentrations of TCDD, but the TCDD bound receptor did not appreciably retard 32P-labeled XRE-3 as determined by electrophoretic mobility shift assay. Incubation of rat hepatic cytosol with TCDD also increased protein kinase C dependent phosphorylation in a concentration dependent manner as well as retarded the mobility of 32P-labeled XRE far more than that observed with thymic cytosol. The ability of the Ah receptor to bind to XRE was temperature dependent. Incubation of thymic cytosol either with okadaic acid or with sodium fluoride did not increase the binding of the Ah receptor to XRE. On mixing hepatic cytosol with thymic cytosol prior to the electrophoretic mobility shift assay, the binding of the hepatic Ah receptor to XRE was reduced in a concentration-dependent manner. A similar effect was observed when renal cytosol was used instead of hepatic cytosol. Thymic cytosol also reduced the binding of the hepatic glucocorticoid receptor to its cognate responsive element. Thymic cytosol did not alter the topology/integrity of XRE. These data suggest that the thymus contains a factor(s) which interferes with the binding of AhR to its cognate responsive element.

Animals↗
PubMed1993

Non-radioactive detection of Mycobacterium tuberculosis LCR products in a microtitre plate format.

As part of the development of the ligase chain reaction (LCR) into a tool which can be used by a wide variety of researchers, we have investigated several analytical detection systems for the products of this amplification reaction. While early work with this technology has used gel electrophoresis to separate the LCR probes from the ligated product, solid phase capture techniques are also applicable, particularly when one of the probes is modified with a 'hook' such as biotin, and the adjoining probe modified with a detectable label. In this study we report a comparison of eight different non-radioactive detection techniques and discuss the analytical sensitivity of each. Detection with laser scanning fluorescent gel electrophoresis remains the most sensitive, with the assay described herein capable of detecting 100 molecules of the Mycobacterium tuberculosis insertion element IS6110 in a background of 4 micrograms of unrelated DNA. This method was followed closely by solid-phase capture and chemiluminescence detection which gave a sensitivity of 1000 molecules of IS6110. Fluorescence detection was approximately 10-fold less sensitive than chemiluminescence detection, and absorbance detection was a further 10-fold less sensitive than fluorescence detection. However, absorbance detection even at this level can still be useful for systems where visual interpretation is desired.

Alkaline Phosphatase↗
PubMed1986

Alpha-emitting, hot particles in Irish sea sediments.

Alpha-emitting, hot particles have been observed in samples of liquid effluent from the nuclear fuel reprocessing plant at Sellafield, U.K. Similar particles, thought to originate from Sellafield, have been found in environmental samples from sites near the plant. The occurrence and distribution of hot particles in surface sediments from the northeastern sector of the Irish Sea are described and the implications for predicting transuranium element behaviour are discussed.

Alpha Particles↗