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[Enhanced formation of plaque in RNA phages/bacteria systems under the effect of surface-active preparations].

The effect of 21 surfactants on the plaque formation was tested in three RNA-phages/Escherichia coli systems and in one RNA-phage/Pseudomonas aeruginosa system. Especially anionic detergents proved to be able to influence the plaque formation substantially. In high concentrations of Metaupon, Fekunil 602, Fekunil S-BA, Emulgator W 270, and Emulgator O-BA the plaque formation by the phages M 12 and f2 was inhibited and in low concentrations it was promoted. In the system Q beta/E. coli AB 301 the effect of the detergents mentioned was restricted to the prevention of the appearance of plaques. The detergent E 30 brought about only plaque inhibitions, too, in all used RNA-phages/E. coli systems. The treatment with effective detergents in the system PP7/P. aeruginosa, however, increased only the number of plaques. This phenomenon was evident in the formation of radiate plaque patterns in the lawn of bacteria. In the case of RNA-phages of E. coli the formation of such trains of lysis depends on the choice of the nutritive medium. The addition of the detergent Fekunil 602 at different times after the contact between phages and host bacteria affects the length of the beams of lysis. The ionogenity of surfactants seems to be of importance for the formation of radiate plaque patterns, since the tested cationic compound and the nonionic surfactants, contrary to anionic surfactants, did not cause any beams of lysis.

Coliphages↗

Nonradiative and radiative deactivation of singlet molecular oxygen (O2(a1deltag)) in micellar media and microemulsions.

The effects of microheterogeneous media (micelles and microemulsions) on the lifetime and, to our knowledge for the first time, on the emission of singlet molecular oxygen (O2 (a1Ag), denoted as 1O2) were investigated. Micellar media and various types of microemulsions based on anionic (sodiumdodecyl sulfate), cationic (cetyltrimethylammonium chloride) or nonionic (Triton X-100) surfactants were formulated for this purpose. The nonradiative and radiative deactivation rate constants (k(d) and k(e), respectively) were determined in selected microheterogeneous media and in the pure solvents used for their formulation, by combining steady-state and time-resolved 1O2, luminescence detection techniques. We have shown that a simple additive model, as used in homogeneous mixtures of solvents, was inadequate for predicting values of k(d) and k(e) in organized media. In contrast, both 1O2 lifetimes (taudelta = 1/k(d)) and k(e) in the microheterogeneous systems investigated could be predicted with good precision from the composition of the media and the taudelta and k(e) values in the pure solvents, using a two-pseudophase kinetic model for the 1O2 distribution. Such a model takes into account the average times spent by 1O2 in the aqueous and lipophilic pseudo-phases of the organized media, the corresponding equilibrium constant (Keq) depending on the nature of the system.

Cetrimonium↗

Crystallization and preliminary x-ray diffraction study of synthetic apolipoprotein E fragment (residues 129-169).

Apolipoprotein E is a plasma protein comprised of a lipid binding region (which together with other apoproteins maintains the structure of lipoprotein particles) and a receptor binding domain (which interacts with cellular receptors for control of triglyceride and cholesterol metabolism). A peptide, comprising residues 129-169 of human apolipoprotein E, which contains both a putative lipid-binding region and receptor binding domain, has been synthesized by solid phase techniques. Diffraction quality crystals of the synthetic apolipoprotein E fragment129-169 have been obtained at room temperature by vapor diffusion with polyethylene glycol in the presence of the nonionic detergent beta-octylglucoside. The crystals have been characterized with x-radiation as orthorhombic, space group I222 or I2(1)2(1)2(1), with unit cell dimensions a = 61.91, b = 30.84, and c = 42.79 A. There are eight molecules per unit cell, with one molecule (Mr = 4771) in each asymmetric unit. Precession photographs show that crystals diffract beyond 2.7-A resolution and are stable in the x-ray beam at room temperature for at least 200 h; thus, they can be used to collect three-dimensional data for a detailed crystallographic analysis.

Apolipoproteins E↗

Crystallization and preliminary x-ray diffraction study of cholera toxin B-subunit.

Cholera toxin binds to its ganglioside GM1 receptor via its B-subunit, a pentameric assembly of identical subunits (Mr = 11,600). Diffraction quality crystals of cholera toxin B-subunit have been obtained at room temperature by vapor diffusion with polyethylene glycol in the presence of the nonionic detergent beta-octyl glucoside. The crystals have been characterized with x-radiation as monoclinic, space group P21, with unit cell dimensions a = 39.0 A, b = 94.3 A, c = 67.5 A, beta = 96.0 degrees. There are two molecules per unit cell, with one molecule (Mr = 58,000) in each asymmetric unit. Precession photographs (micron = 13 degrees) show that crystals diffract beyond 3.3-A resolution and are stable in the x-ray beam at room temperature for at least 40 h; thus, they can be used to collect three-dimensional crystallographic data.

Cholera Toxin↗

Contrast enhancement of brain tumors and irradiated normal brain: a comparison of iohexol and iothalamate.

The nonionic contrast agent iohexol was compared with the ionic agent iothalamate for contrast enhancement of brain tumors and radiation brain damage in dogs. Tissue enhancement during infusion of contrast and 5, 10, 15 and 30 min later was measured using quantitative computed tomography. Blood iodine was measured using x-ray fluorescence. Peak contrast enhancement occurred during infusion in tumors and after 5 min in irradiated normal brain for both contrast agents. Absolute uptake of contrast in each lesion was the same for both agents when normalized to total grams of iodine administered. Blood iodine levels were slightly but not significantly higher using iohexol. The reduced osmotic load and similar contrast enhancement of lesions suggest that the nonionic contrast medium iohexol may be a useful agent for routine CT of the brain.

Animals↗

Evidence relating cessation of respiration, cell envelope changes, and death in ultraviolet-irradiated Escherichia coli B-r cells.

Ionic and nonionic detergents have little effect on respiring bacteria, but in cultures poisoned with KCN rapid solubilization of the cell membrane, as indicated by turbidity losses, takes place. Ultraviolet radiations cause Escherichia coli cells grown in minimal medium with glycerol as a carbon source to cease respiring and growing about 1 h after irradiation. We tested the effect of the nonionic detergent Triton X-100 on growth and cell membrane dissolution (both measured by turbidity changes), respiration, and viability of unirradiated and irradiated E. coli B/r cells. When the detergent was added to cells immediately after irradiation, a decrease in turbidity occurred only when respiration was about to cease; when it was added after cessation of respiration, the turbidity loss was immediate. In both cases the turbidity loss was about 60%, and disintegration of the cell walls did not take place. 5-Fluorouracil (FU) and thermal (42 C) treatments cause respiration of irradiated cells to be maintained and also cause viability increases. Irradiated cells treated with FU and detergent show no turbidity loss just prior to the time respiration normally ceases, but a loss does occur in irradiated cells incubated with detergent at 42 C. We conclude that FU maintains respiration for all of the cells, but that thermal treatment maintains respiration for only part of the cells. In all cases the detergent had only a negligible effect on the respiration and viability of unirradiated and irradiated cells. We conclude that Triton X-100 causes solubilization of cell membranes of only nonrespiring cells that are not destined to survive.

Anaerobiosis↗

The effect of intravenous contrast on intensity-modulated radiation therapy dose calculations for head and neck cancer.

OBJECTIVE: The objective of this study was to determine if the use of intravenous contrast results in clinically important errors in intensity-modulated radiation therapy (IMRT) dose calculations for head and neck radiotherapy treatment planning. MATERIALS AND METHODS: Nonionic, iodinated intravenous contrast (Iohexol) was administered during the treatment planning computed tomography (CT) scan of 5 patients with head and neck cancer of varying disease sites. The potential effect of intravenous contrast was studied by changing the density of the contrast-enhanced vessels. An inverse IMRT plan was generated from an unmanipulated "normal contrast" planning scan. We then applied the same planning parameters to a "no contrast" planning scan. The effect of intravenous contrast was quantified by calculating the percent change of dose in a variety of target and normal structures. To evaluate a worst-case scenario situation, this comparison was repeated by assigning the vessels the maximum density in our planning system ("maximum contrast" density plan). RESULTS: Dose differences between a planning set of images using intravenous contrast and a set of images without contrast were less than 0.2% for all relevant target volumes and critical structures. A worst-case scenario in which normal contrast was overridden with "maximum contrast" led to small dose differences, generally less than 0.5%. CONCLUSIONS: Planning head and neck IMRT from CT scans that contain intravenous contrast does not result in clinically important errors in dose delivery.

Contrast Media↗

Polyethyleneimine grafted with pluronic P85 enhances Ku86 antisense delivery and the ionizing radiation treatment efficacy in vivo.

In an effort to improve the efficacy of antisense delivery, we evaluated polyethyleneimine (PEI, 2 kDa) alone or grafted with nonionic amphiphilic block copolymer Pluronic (P85) as a carrier for Ku86 antisense oligonucleotide (ASO) delivery. Ku86 is an abundant nuclear protein that plays an important role in nonhomologous DNA end joining and has implications in tumorigenesis and acquired drug resistance. Transfection of adherent and suspension cell lines with Ku86 ASOs complexed with P85-g-PEI (2 kDa) conjugates was associated with a specific decrease in Ku86 mRNA levels (EC50<75 nM and EC50<250 nM, respectively, n=3). More importantly, no requirement for reduced serum conditions was necessary during transfection. In contrast, whereas Ku86 ASOs complexed with PEI (2 kDa) alone were effective in decreasing Ku86 mRNA levels in adherent cell lines (EC50<75 nM, n=3), the formulation did not produce any detectable decrease in Ku86 mRNA levels in suspension cell lines. Transfection of adherent cell lines with 500 nM Ku86 ASOs formulated with P85-g-PEI (2 kDa) was associated with a specific decrease (<10% remaining of control) in Ku86 protein expression and a two-fold increased cell death after treatment with ionizing radiation (IR). In athymic nude mice bearing subcutaneous human HT29 colon adenocarcinoma xenografts, Ku86 ASO-P85-g-PEI (2 kDa) administration (15 mg/kg, subcutaneously) with a Q1D x 7 treatment schedule, when combined with a single dose of IR (6 Gy), caused a significant inhibition of HT29 tumor growth compared with mismatch- and naked antisense-pretreated control groups (time from 200 to 1000 mm3, 126.9 versus 84.18 and 87.76 days, P<0.005). A potentiation of the antitumor activity was observed in all mice treated with Ku86 ASO-P85-g-PEI (2 kDa) formulation; however, tumor growth inhibition was reversible upon treatment cessation. No morbidity/mortality or changes in histopathology were observed under this treatment regiment. Our results indicate that P85-g-PEI (2 kDa) conjugates may increase the efficacy of Ku86 ASO delivery in management of resistant malignancies, thus providing a rationale for their evaluation in cancer patients in combination with conventional anticancer therapies.

Animals↗

Change in patient doses from radiological examinations at the Vancouver General Hospital, 1991-2002.

OBJECTIVE: Much concern has been expressed over the radiation doses and potential harm from x-ray examinations. However, there have been few longitudinal studies in North America. A survey of doses from radiological examinations in Canada was last carried out in 1995. This study was undertaken to estimate the change in the number of patient examinations and patient dose since this last Canadian survey. METHODS: The number of radiological examinations and numbers of patients for the years 1991 to 2002 were obtained from workload statistics, which are reported to the Canadian government each year. Radiological examinations were of the following type: general, gastrointestinal or genitourinary, angiography, and computed tomography (CT). Average doses were calculated for each group of examinations. RESULTS: From 1991 to 2002 there was an increase of 28% in the total number of x-ray examinations performed. The proportion of most types of examination has stayed fairly constant, except for CT, which has increased fourfold in the last 8 years. The striking change is the increased contribution to patient effective dose from CT since 1996, these examinations now comprising nearly 60% of the total patient dose. The average annual effective dose per patient has nearly doubled-from 3.3 mSv in 1991 to 6.0 mSv in 2002. CONCLUSION: This paper provides a simple method for any Canadian hospital to estimate the radiation dose to its patients. At the Vancouver General Hospital (VGH), the number of patient examinations has increased by 28%, but the average annual patient effective dose has almost doubled. CT is now by far the largest contributor to patient dose in diagnostic radiology. Efforts need to be made to reduce patient dose by such methods as reduction in unnecessary exams, substitution of nonionizing techniques where possible, and optimization of dose.

British Columbia↗

X-ray mammography and diaphanography in screening for breast cancer.

Modern low-dose dedicated x-ray mammography has been demonstrated in large clinical trials to have high efficacy in the detection of occult breast malignancy. Such early detection results in decreased fatality rates. A theoretical risk of breast cancer induced from diagnostic ionizing radiation has not been proven for the low doses now used in mammographic techniques. High accuracy can be achieved in the mammographic evaluation of breast lesions. In contrast, nonionizing transillumination of the breast, or diaphanography, remains an investigational approach to the evaluation of breast disease and warrants further controlled study before its widespread clinical application.

Adult↗

Electronic thermography in the assessment of internal derangement of the temporomandibular joint. A pilot study.

Current diagnostic imaging methods to evaluate internal derangement of the temporomandibular joint (TMJ) include arthrotomography, arthroscopy, computed tomography, and magnetic resonance imaging. Most of these techniques either require ionizing radiation or are invasive, and all are expensive. Electronic thermography (ET) is an alternative diagnostic imaging modality that is inexpensive, nonionizing, and noninvasive. The purpose of this pilot study was to assess ET in the diagnosis of internal derangements of the TMJ. The study population consisted of 11 patients with internal derangement, and 12 normal patients. ET was conducted with an infrared thermographic unit, at 1.0 degrees and 0.5 degrees C sensitivity. Results from subjective blinded interpretations by two experts for the diagnosis of internal derangement of the TMJ were as follows: sensitivity, 86% (+/- 7.8%); specificity, 78% (+/- 7.1%). Results from objective measurements of thermal symmetry of the TMJ region were as follows: normal subjects, 89.3% (+/- 3.0%); internal derangement patients, 66.1% (+/- 16.2%); t21 = -4.89, p less than 0.01. In conclusion, ET appears to have some promise as a diagnostic aid in the evaluation of internal derangement of the TMJ. Additional, more extensive studies are needed before thermographic diagnosis of internal derangement is accepted clinically.

Adult↗

Target size analysis of prostaglandin endoperoxide synthase. Radiation inactivation of both cyclooxygenase and peroxidase correlated with the monomer of 72 kDa.

To determine the size of the functional catalytic unit of prostaglandin endoperoxide (prostaglandin H) synthase, radiation inactivation experiments were performed. Both microsomes from ovine seminal vesicles and purified enzyme were irradiated with 10 MeV electrons. The enzymic activities of prostaglandin H synthase, cyclooxygenase and peroxidase, showed mono-exponential inactivation curves dependent on radiation dose, indicating molecular masses of approximately 72 kDa. The enzyme in microsomes, in its native environment, as well as in its purified state after solubilisation with nonionic detergent showed identical molecular masses. The results clearly demonstrate that the monomer of the enzyme with an apparent molecular mass of 72 kDa (SDS/PAGE) is the functional unit for catalysis of both activities. Hence the two active sites of cyclooxygenase and peroxidase reside on the same polypeptide chain.

Animals↗

The influence of chromatin structure on the frequency of radiation-induced DNA strand breaks: a study using nuclear and nucleoid monolayers.

To assess the influence of chromatin structure on the frequency of radiation-induced DNA strand breaks, the alkaline unwinding technique was applied to nuclear and nucleoid monolayers. These chromatin substrates were prepared by treating human fibroblasts grown as monolayers with the nonionic detergent Triton X-100 and varying concentrations of cations. The chromatin structure was modified either by a stepwise removal of DNA-bound proteins by extraction in increasing concentrations of monovalent salt, or by the addition or deletion of mono- and divalent cations to condense or decondense the chromatin, respectively. It was found that the stepwise removal of DNA-bound proteins from the chromatin dramatically increased the frequency of radiation-induced DNA strand breaks. The DNA-bound proteins showed a qualitative difference in their ability to protect the DNA where proteins removed by salt concentrations above 1.0 M exerted the greatest protection. Furthermore, the frequency of radiation-induced DNA strand breaks was found to be 6 times lower in condensed chromatin than in decondensed chromatin and about 80 times lower than in protein-depleted chromatin. It is concluded that the presence of DNA-bound proteins and the folding of the chromatin into higher-order structures protect the DNA against radiation-induced strand breaks.

Cesium Radioisotopes↗

Elimination kinetics of iopamidol, a new water soluble nonionic radiographic contrast medium, analyzed by radioactivation.

We have studied the elimination kinetics of iopamidol employing radioactivation and radiochemical separation. This method offers the advantage of guaranteeing absolutely no interference by radiation with tissue distribution or elimination kinetics of the analyzed compound. Our results show that iopamidol does not cross the blood-brain barrier, has no effect on thyroid iodine uptake and does not accumulate in the liver.

Animals↗

Predicting the effect of nonionic surfactants on dispersed droplet radii in submicron oil-in-water emulsions.

A novel theoretical model which describes the mass mean radius of oil droplets in an oil-in-water emulsion is described. A modified form of the Langmuir adsorption isotherm is used to account for nonlinear adsorption of surfactant to an oil-water film and its effect on interfacial tension and oil droplet radius. On the basis of this model, the mass mean oil droplet radius may be related to bulk surfactant concentration for a nonionic surfactant. An analysis of the mineral oil-Triton X405-water system shows that aqueous solutions of Triton X405 against mineral oil behaved in accord with the proposed model. gamma m, the maximum interfacial tension lowering by Triton X405, was estimated to be 20.9 dyne/cm. A surfactant specific apparent constant (B) which relates the rate constant for adsorption to the interface to the rate constant for desorption from the interface into the aqueous bulk was estimated as 5.44E4 cm3/g. The pressure across the curved interface, delta P, was estimated as 1.05E6 dyne/cm2. The theoretical model appears to be consistent with experimentally observed oil droplet radii and is considered to be an accurate representation of the mechanics of dispersed droplet radii under conditions of moderate load of nonionic surfactant.

Emulsions↗

Micro CT and Micro MR imaging of 3D architecture of animal skeleton.

Quantitative assessment of three-dimensional (3D) trabecular structural characteristics may improve our ability to understand the pathophysiology of osteoporosis, to test the efficacy of pharmaceutical intervention, and to estimate bone biomechanical properties. Considerable progress has been made in advanced imaging techniques for noninvasive and/or nondestructive assessment of 3D trabecular structure and connectivity. Micro computed tomography (microCT) has been used to measure 3D trabecular bone structure in rats, both in vivo and in vitro. It can directly quantify 3D trabecular bone structure such as trabecular volume, trabecular thickness, number, separation, structure model index, degree of anisotropy, and connectivity, in a model-independent manner. We have used microCT to study ovariectomy (OVX) induced osteopenia in rats and its treatment with agents such as estrogen, and sodium fluoride. We have demonstrated that 3D microCT can quantify mouse trabecular and cortical bone structure with an isotropic resolution of 9 microm(3). It is also useful for studying osteoporosis in mice and in phenotypes of transgenic mice or gene knockout mice. MicroCT can be used to quantify osteogenesis in mouse Ilizarov leg lengthening procedures, to quantify osteoconduction in a rat cranial defect model, and to quantify cortical bone porosity. Recently, microCT using high intensity and tight collimation synchrotron radiation to achieve spatial resolution of 1-2 microm has provided the capability to assess additional features such as resorption cavities. Unlike microCT, micro magnetic resonance imaging (IMRI) is nonionizing. Recently, the ability of microMRI to assess osteoporosis in animal models has been explored. Using a small, high-efficiency coil in a high-field imager, microMRI can give resolutions sufficient to discriminate individual trabeculae. We have shown that, with appropriate settings, it is possible to image trabecular bone in rats in vivo and in vitro. In our study of OVX rats, analysis of microMR images can demonstrate differences in rat trabecular bone that are not detected by DXA measurements. In a rabbit OA model, with the OA induced by meniscectomy or anterior cruciate ligament transection, MRI shows decreased cartilage thickness, subchondral osteosclerosis and osteophytes, while radiographs can only show subchondral osteosclerosis and osteophytes could not be found. Advanced imaging methods are able to measure 3D trabecular structure and connectivity in arbitrary orientations in a highly automated, objective, non-user-specific manner, allowing greater numbers of samples for unbiased comparisons between controls and the disordered or treated. They can be utilized on a large sample leading to fewer sampling errors. They are non-destructive allowing multiple tests such as biomechanical testing and chemical analysis on the same sample; and non-invasive permitting longitudinal studies and reducing the number of animals needed.

Journal Article↗

Fluorescence and dynamic light scattering studies of niosomes-membrane mimetic systems.

Niosomal vesicles are more stable than liposomal vesicles due to higher chemical stability of surfactants compared to phospholipids. Niosomes have been prepared from Span20, Span80, Tween20 and Tween80. Fluorescence resonance energy transfer studies have been performed in these systems to determine donor-acceptor distances. It has been found that the fluorescence resonance energy transfer efficiency is better in niosomes compared to micelles. The formation of niosomes is guided by the hydrophile-lipophile balance value of the nonionic surfactant.

Cholesterol↗