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B Vojnovic

Publications and source records attributed to B Vojnovic.

At least 37 records · Page 2Linked to original sources
PubMed1997

A charged-particle microbeam: I. Development of an experimental system for targeting cells individually with counted particles.

Charged-particle microbeams provide a unique opportunity to control precisely, the dose to individual cells and the localization of dose within the cell. The Gray Laboratory is now routinely operating a charged-particle microbeam capable of delivering targeted and counted particles to individual cells, at a dose-rate sufficient to permit a number of single-cell assays of radiation damage to be implemented. By this means, it is possible to study a number of important radiobiological processes in ways that cannot be achieved using conventional methods. This report describes the rationale, development and current capabilities of the Gray Laboratory microbeam.

DNA↗
PubMed1997

A charged-particle microbeam: II. A single-particle micro-collimation and detection system.

The use of a charged-particle microbeam provides a unique opportunity to control precisely, the number of particles traversing individual cells and the localization of dose within the cell. The accuracy of 'aiming' and of delivering a precise number of particles crucially depends on the design and implementation of the collimation and detection system. This report describes the methods available for collimating and detecting energetic particles in the context of a radiobiological microbeam. The arrangement developed at the Gray Laboratory uses either a 'V'-groove or a thick-walled glass capillary to achieve 2-5 microns spatial resolution. The particle detection system uses an 18 microns thick transmission scintillator and photomultiplier tube to detect particles with > 99% efficiency.

Particle Accelerators↗
PubMed1997

Measurement of tumor oxygenation: a comparison between polarographic needle electrodes and a time-resolved luminescence-based optical sensor.

A novel oxygen sensor which does not rely on electrochemical reduction has been used to measure the oxygenation of the murine sarcoma F in a comparative study with an existing polarographic electrode that is available commercially. The prototype luminescence sensor yielded an oxygen distribution comparable with readings made using a pO2 histograph. The percentage of regions detected that had a pO2 less than 5 mm Hg was 79 and 75 using the Eppendorf pO2 histograph and the luminescence fiber optic sensor, respectively. These values were compatible with a measured radiobiologically hypoxic fraction of 67% in this tumor. The polarographic method detected more regions with a pO2 of 2.5 mm Hg or less (69%) compared with the optical sensor (50%) (P < 0.05). This could reflect differences in the oxygen use of the sensing devices. This initial assessment indicates the potential of a fiber-optic-based oxygen-monitoring system. Such a system should have several advantages including monitoring temporal oxygen changes in a given microregion and use with NMR procedures.

Animals↗
PubMed1997

Post-traumatic AV fistulas and pseudoaneurysms.

METHODS: The authors present the surgical treatment of 20 post-traumatic arteriovenous fistulas and 33 arterial pseudoaneurysms that have been treated in the last 5 years in the Centre for Vascular Surgery of the Institute for Cardiovascular Diseases, Clinical Centre of Serbia in Belgrade. Five women and 45 men (mean age 31.7 years) were examined. There were 28 war and 22 non-combatant injuries. In most cases superficial femoral artery and vein were involved. The average time elapsed from the moment of injury until the operation started, was 9 months in patients with AV fistulas, and one month for patients with pseudoaneurysms. RESULTS: In all of the patients with AV fistulas, arterial and venous reconstructions were performed, except in 4 cases where the veins were ligated. Surgical reconstruction was performed in 26 patients with pseudoaneurysms, while in 7 cases the artery was ligated. There were no cases of postoperative ischemia in patients due to arterial ligation. Patients were followed for 2 years and 2 months postoperatively. As far as the reconstructive operations are concerned, the postoperative patency rate was 100%, while limb salvage was achieved in 96.9%. Namely, one amputation was done in spite of high arterial patency rate, which was indicated by massive bone-muscle tissue loss, occurring during mine explosive injury. CONCLUSIONS: Because of the rapid disease progress, the authors suggest that the operative treatment of post-traumatic AV fistulas and pseudoaneurysms should be performed as soon as possible. This was supported by good follow-up results in operatively treated patients.

Adolescent↗
PubMed1996

Radiosensitisation in normal tissues with oxygen, carbogen or nicotinamide: therapeutic gain comparisons for fractionated x-ray schedules.

METHODS: Radiosensitisation with oxygen, carbogen or nicotinamide alone and oxygen or carbogen combined with nicotinamide was compared in early and late responding normal tissues in rodents. X-ray treatments were delivered as single doses or fractionated schedules of 2 fractions in 1 day, 2, 12 and 36 fractions in an overall time of 12 days and 10 fractions in 5 or 12 days. Acute skin reactions, survival of intestinal crypts, breathing rate, reduction in the packed red-cell volume and clearance of 51Cr-EDTA were used as assays of epidermal, gut, lung and renal damage. RESULTS: Relative to air-breathing mice, carbogen or oxygen produced a small, and not always significant, increase in sensitivity (enhancement ratios < or = 1.15) in gut, lung and kidneys; however, in skin a dose enhancement of 1.2-1.3 was observed. The effect of nicotinamide in air, carbogen or oxygen was studied only in lung and gut. The drug produced variable but generally significant increases in radiosensitisation ( < or = 1.26) in all three gases. Relative to treatments in air, enhancement ratios for nicotinamide alone were usually slightly higher than those observed when either carbogen or oxygen were administered without the drug. With all three modifiers (i.e. oxygen, carbogen, nicotinamide alone or for the drug-gas combinations) there was no significant change in the enhancement ratios observed as the number of radiation dose fractions was varied. CONCLUSIONS: Comparisons with fractionated X-ray studies done previously in rodent tumours indicate that a therapeutic benefit, relative to lung, gut and renal damage, would be observed with oxygen or carbogen alone but not with nicotinamide alone. The greatest gain would be achieved with the combination of carbogen and nicotinamide, with which a benefit was observed even relative to epidermal damage. These results indicate that some decrease in normal tissue tolerance could be observed when using these modifiers in clinical radiotherapy and, although small, the appropriate dose reductions should be considered; caution should be exercised especially when carbogen and nicotinamide are used in conjunction with the more radical accelerated schedules.

Animals↗
PubMed1996

Inactivation of V79 cells by low-energy protons, deuterons and helium-3 ions.

Previous work by ourselves and by others has demonstrated that protons with a linear energy transfer (LET) about 30 keVmum(-1)are more effective at killing cells than doubly charged particles of the same LET. In this work we show that by using deuterons, which have about twice the range of protons with the same LET, it is possible to extend measurements of the RBE of singly charged particles to higher LET (up to 50 keVmum(-1). We report the design and use of a new arrangement for irradiating V79 mammalian cells. Cell survival measurements have been made using protons in the energy range 1.0-3.7 MeV, deuterons in the energy range 0.9-3.4MeV and 3He2+ ions in the energy range 3.4-6.9 MeV. This corresponds to volume-averaged LET (within the cell nucleus) between 10 and 28 keVmum(-1) for protons, 18-50 keVmum(-1) for deuterons, and 59-106 keVmum(-1) for helium ions. Our results show no difference in the effectiveness of protons and deuterons matched for LET. However, for LET above about 30 keVmum(-1) singly charged particles are more effective at inactivating cells than doubly-charged particles of the same LET and that this difference can be understood in terms of the radial dose distribution around the primary ion track.

Animals↗
PubMed1996

Measurement of oxygen tension in tumours by time-resolved fluorescence.

Tumour oxygenation is important in clinical radiotherapy because hypoxic cells are radioresistant. Knowledge of the state of tumour oxygenation would be advantageous for maximising effectiveness of treatment. A prototype fibre optic fluorosensor for measuring low (radiobiologically relevant) levels of oxygen is described. Based on oxygen quenching of the fluorescence of an excited fluorophor immobilised in a polymer at the end of an optical fibre, the sensor shows promise in overcoming some of the limitations of existing oxygen sensor systems. The prototype fibre optic sensor operates most effectively in the 0-2% oxygen range with fast response and settling times. Preliminary results from measurements in tumours are presented.

Animals↗
PubMed1994

Action spectra for single- and double-strand break induction in plasmid DNA: studies using synchrotron radiation.

Ionizing radiations deposit a wide range of energies in and around DNA and this leads to a corresponding spectrum of complexity of the lesions induced. The relationships between the amount of energy deposited and the yields and types of damage induced are important in modelling the physical and chemical stages of radiation effect and linking them to biological outcome. To study these relationships experimentally, plasmids were mounted as a monolayer and exposed in vacuum to near-monoenergetic photons from the Daresbury Synchrotron. After irradiation, the DNA was washed off and assayed for single-(ssb) and double-strand breaks (dsb) using agarose gel electrophoresis. Dose-effect relationships for ssb and dsb induction were obtained at various energies in the range 8-25 eV. The initial responses in the low-dose region allowed damage yields to be estimated. However, a common feature is that the responses showed energy-dependent plateaus at higher doses as if a fraction of the DNA were shielded. Various measures were taken both to minimize and to correct for this effect. The data appear to show that the yields of ssb and dsb increase only slowly with photon energies > 10 eV, with a suggestion of similar threshold energies for both lesions. In the energy range covered, the yield of ssb is 12-20-fold greater than that of dsb. The data indicate that ssb and dsb may have a common precursor in this system. Earlier work with low-energy electrons showed that at 25 eV ssb were induced but no dsb were detected.

DNA↗
PubMed1993

Measurement of DNA damage by electrons with energies between 25 and 4000 eV.

All ionizing radiations deposit energy stochastically along their tracks. The resulting distribution of energies deposited in a small target such as the DNA helix leads to a corresponding spectrum in the severity of damage produced. So far, most information about the probable spectra of DNA lesion complexity has come from Monte Carlo studies which endeavour to model the relationship between the energy deposited in DNA and the damage induced. The aim of this paper is to establish methods of determining this relationship by irradiating pBR322 plasmid DNA using low energy electrons with energies comparable with the minimum energy thought to produce critical damage. The technique of agarose gel electrophoresis has been used to ascertain the fraction of DNA single- and double-strand breaks induced by monoenergetic electrons with energies as low as 25 eV. Our data show that the threshold electron energy for induction of single-strand breaks is < 25 eV, and for double-strand breaks between 25 and 50 eV.

DNA↗
PubMed1991

Absorption spectra of radical forms of 2,4-dihydroxybenzoic acid, a substrate for p-hydroxybenzoate hydroxylase.

Combined optical and conductimetric measurements in aqueous solution indicate that at high pH (greater than or equal to 10).OH radicals react with the phenoxide form of 2,4-dihydroxybenzoic acid to form transiently phenoxyl radicals and a small amount of hydroxyeyclohexadienyl (HCHD) radicals by 150 ns. The respective yields of 88 and 12% of the total.OH radical yield were deduced from conductance and optical changes as well as from studies using a low potential reductant. The HCHD radical possesses a pKa of 8.0 +/- 0.1 and the constructed spectrum of the deprotonated forms of HCHD has a lambda max at 420 nm with a minimum extinction coefficient of approximately 7250 M-1 cm-1. The red shift in lambda max and increase in extinction coefficient compared to the revised spectral properties of the protonated form of the HCHD radical (lambda max at 390 nm with extinction coefficient of approximately 4500 M-1 cm-1), together with the pKa of the HCHD radical, provide an explanation for the pH-dependent spectral changes of the so-called highly absorbing intermediate II species, observed in the functioning of the enzyme p-hydroxybenzoate hydroxylase. These results add further to the evidence in support of the proposal that intermediate II is composed of species which absorb similarly to the flavin 4(a)-hydroxide and a form of the substrate/product such as the HCHD radical (Anderson, R. F., Patel, K. B., and Stratford, M. R. L. (1987) J. Biol. Chem. 262, 17475-17479).

4-Hydroxybenzoate-3-Monooxygenase↗
PubMed1991

Determination of X-ray-induced damage to the murine colon using tissue compliance measurements.

A proctometroscope has been developed to measure the mechanical functioning of the colon in a murine model. A balloon-tipped probe is inserted into the colorectal region of anaesthetized mice and inflated hydraulically, at a constant rate, by a motor-driven syringe. Balloon pressure and volume are monitored, processed and plotted on a X-Y/t recorder. The balloon is inserted within the irradiated area, such that the centre of the balloon is 1 cm proximal to the anus, and inflated up to either a maximum volume of 100 microliters or a maximum pressure of 100 cmH2O, depending on which occurs first. Compliance (delta V/delta P) of the colon was measured at various intervals following X-irradiation, a dose-dependent decrease being observed at 24 weeks. This decrease was progressive with time out to 72 weeks postirradiation. The compliance measured, in both control and irradiated groups, was not dependent on the inflation rate used to make the determination, and no iatrogenic effects have been detected after repeated probing during a long-term study. The use of different anaesthetic agents was also investigated, no significant difference between the compliance of mice anaesthetized with the different agents being detected. Use of this technique provides fully quantitative data on the function of the colon following radiation injury, and provides an alternative to other physiological assays. The technique is non-destructive, rapid, easy to use and non-invasive to the tissue, thus the onset and progression of damage can be followed in each mouse over long postirradiation periods.

Animals↗
PubMed1991

The effects of counter-ion condensation and co-ion depletion upon the rates of chemical repair of poly(U) radicals by thiols.

Bimolecular rate constants for reactions of poly(U) radicals with a series of thiols of varying net charge (Z) were measured by pulse radiolysis with conductivity detection at low ionic strength. At pH 7 and 18 degrees C the values of k2 (M-1s-1) were: reduced glutathione (Z = -1), less than 500; 2-mercaptoethanesulphonic acid (Z = -1), 1.5 x 10(3); 2-mercaptoethanol (Z = 0), 1.8 x 10(5); cysteine (Z = 0), 2.0 x 10(5); cysteamine (Z = +1), 4.1 x 10(7). Values determined at pH 4 were: 2-mercaptoethanol, 6.1 x 10(5); cysteamine 2.2 x 10(8); N-(2-mercaptoethyl)-1,3-diaminopropane (WR-1065, Z = +2), 4.6 x 10(8). The variation in rate with structure could not reasonably be attributed to inherent reactivity differences in the thiols and was ascribed to inhomogeneous distributions of the thiols in solution resulting from electrostatic interactions. Thus, cationic thiols are concentrated approximately 100-fold near poly(U), relative to neutral thiols, as a consequence of counter-ion condensation, whereas anionic thiols have approximately 100-fold lower concentration near poly(U) than neutral thiols as a result of co-ion depletion. These results show that the ability of a thiol to repair radical sites in a polyanion is dramatically influenced by its net charge as a consequence of the counter-ion condensation and co-ion depletion phenomena.

Cysteamine↗
PubMed1989

A single-shot rapid-mixing device for radiobiological studies with mammalian cells.

A single-shot rapid-mixing device is described for the rapid addition of solutions of radiation-modifying agents, to cell suspensions, at well-defined times relative to a pulse of radiation. The liquid injection system could be used to initiate or quench a wide range of chemical or biochemical reactions. The rapid-mixing device is based on a syringe driven by a stepper motor and can inject up to 2 cm3 liquid in less than 100 ms. The radiation source, a 4 MV Van de Graaff accelerator, provides an electron beam which is deflected from the beam dump on to the sample in two stages, providing a 10 ms radiation pulse. A digital delay circuit defines the interval between mixing and irradiation. The apparatus has been designed to study the kinetics of processes that occur over a time range extending from about 0.1 s to some minutes. It bridges the gap between the ranges available with conventional fast-mixing and those using standard X- or gamma-irradiation methods. The time resolution of the technique has been examined by following the timecourse of radiosensitization by oxygen in mammalian cells. The timecourse of radioprotection of aerobic mammalian cells by dithiothreitol has been measured using the technique.

Animals↗
PubMed1989

The irradiation of V79 mammalian cells by protons with energies below 2 MeV. Part I: Experimental arrangement and measurements of cell survival.

The relative biological effectiveness (RBE) has been determined for protons with mean energies of 1.9, 1.15 and 0.76 MeV, from measurements of the survival of V79 Chinese hamster cells. The cells are supported as a monolayer and are swept through a beam of scattered protons produced using a 4 MeV Van de Graaff accelerator. An estimation of the dose and unrestricted linear energy transfer (LET) variation within the sensitive volume of the cells is given for the three proton energies. The RBEs for cell survival (relative to 250 kVp X-rays) at the 10 per cent survival level are 1.6, 1.9 and 3.36 for protons with track-average LETs of 17, 24 and 32 keV microns-1 respectively, and the data suggest that protons are most effective at about 40-50 keV microns-1. It is shown that the proton RBEs can be reconciled with those of other light ions if plotted against z*2/beta 2 (where z* is the effective charge and beta is the relative velocity) rather than against LET.

Animals↗
PubMed1987

Fast protonation of adenosine and of its radical anion formed by hydrated electron attack; a nanosecond optical and dc-conductivity pulse radiolysis study.

The study of the reaction of the hydrated electron with adenosine by optical and dc-conductivity pulse radiolysis on nano- and microsecond timescales has been carried out in an attempt to answer the question whether the electron adduct radical becomes protonated or not. The following conclusions have been reached: (1) the reaction of the hydrated electron with adenosine is followed by a water-mediated protonation, which must be complete with 5 ns; (2) no spectral indication of a further protonation of the protonated electron adduct of adenosine of 2'-deoxyadenosine has been found between 40 and 5000 ns; (3) the equilibrium reaction between radiation produced H3O+ and adenosine with a pKa of 3.5 plays an important role in the kinetics of the conductivity transients.

Adenosine↗
PubMed1987

An arrangement for irradiating cultured mammalian cells with aluminium characteristic ultrasoft x-rays.

Ultrasoft x-rays are useful for testing the validity of mechanistic models of biological damage caused by radiation. Described here is the construction and operation of a cold-cathode transmission-target discharge tube for irradiating mammalian cells in vitro with aluminium characteristic x-rays (1.487 keV). Particular attention is given to the problems of sample preparation and dosimetry for this shallowly penetrating radiation. The proportion of contaminating bremsstrahlung radiation is measured to establish the optimum operating conditions. Preliminary data from experiments using V79 Chinese hamster cells show that aluminium characteristic x-rays are about twice as effective at inactivating the cells as 250 kVp x-rays.

Aluminum↗