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An altered distribution and elimination of digoxin in anemic patients and experimentally induced anemic rats.

Digoxin was administered orally to eight anemic patients in their anemic and convalescent stages, and serum digoxin concentration was determined by radioimmunoassay. In the anemic patients a significantly lower level of serum digoxin concentration was observed in anemic state compared with convalescent stage at 72 hours after the drug administration (p less than 0.01). In usual clinical use, a full digoxin effect is expected to be attained as 72 hours. Tritiated digoxin was administered intravenously to anemic and control rats and the tritium in samples of the blood, myocardium and urine were counted in a liquid scintillation counter. The anemic rats showed significantly lower level of serum 3H-digoxin at 6 hours (p less than 0.01) and lower myocardial concentration at 24 hours (p less than 0.01). Larger amount of urinary excretion of 3H-digoxin was observed in the anemic rats 6 hours after the drug administration. No significant difference in fecal excretion of 3H-digoxin was found between the anemic and control rats.

Anemia, Hypochromic↗

No radiation protection reasons for restrictions on 14C urea breath tests in children.

Traditional (14)C urea breath tests are normally not used for younger children because the radiation exposure is unknown. High sensitivity accelerator mass spectrometry and an ultra-low amount (440 Bq) of (14)C urea were therefore used both to diagnose Helicobacter pylori (HP) infection in seven children, aged 3-6 years, and to make radiation dose estimates. The activity used was 125 times lower than the amount normally used for older children and 250 times lower than that used for adults. Results were compared with previously reported biokinetic and dosimetric data for adults and older children aged 7-14 years. (14)C activity concentrations in urine and exhaled air per unit administered activity for younger children (3-6 years) correspond well with those for older children (7-14 years). For a child aged 3-6 years who is HP negative, the urinary bladder wall receives the highest absorbed dose, 0.3 mGy MBq(-1). The effective dose is 0.1 mSv MBq(-1) for the 3-year-old child and 0.07 mSv MBq(-1) for the 6-year-old child. For two children, the 10 min and 20 min post-(14)C administration samples of exhaled air showed a significantly higher amount of (14)C activity than for the rest of the children, that is 6% and 19% of administered activity exhaled per hour compared with 0.3-0.9% (mean 0.5%) of administered activity exhaled per hour indicating that these two children that is were HP positive. For a 3-year-old HP positive child, absorbed dose to the urinary bladder wall was 0.3 mGy MBq(-1) and effective dose per unit of administered activity was 0.4 mSv MBq(-1). Using 55 kBq, which is a normal amount for older children when liquid scintillation counters are used for measurement, the effective dose will be approximately 6 micro Sv to a 3-year-old HP negative child and 20 microSv to a HP positive child. Thus there is no reason for restrictions on performing a normal (14)C urea breath test, even on young children.

Adolescent↗

Intercomparison of radiation instruments for cosmic-ray with heavy ion beams at NIRS (ICCHIBAN project).

The first InterComparison for Cosmic-ray with Heavy Ion Beams At NIRS (ICCHIBAN) project is an ongoing, international collaboration organized at the National Institute of Radiological Sciences (NIRS), Japan, for the purpose of characterizing and comparing at a controlled, ground-based heavy ion facility the radiation response of instruments used aboard piloted spacecraft for crew and area dosimetry. We present preliminary results from the first set of ICCHIBAN exposures made at HIMAC heavy ion accelerator in February 2002. The initial series of exposures (1st ICCHIBAN run) was designed to establish the response of active detectors to two well-characterized heavy ion beams; 400 MeV/nucleon 12C and 400 MeV/nucleon 56Fe. These beams are representative in charge and energy of two of the most significant heavy ion components present in the galactic cosmic radiation spectrum. The properties of the incident beam, including intensity, profile, charge and total energy, were characterized using several different detector systems, including silicon detectors, CR-39 plastic nuclear track detectors and plastic scintillation counters. Once the response of each detector to heavy ion beams of known composition has been measured, results from on-orbit measurements made by the different instruments can be more meaningfully compared. We conclude by discussing plans for future ICCHIBAN runs, including next 2nd ICCHIBAN run for passive detectors in early summer 2002.

Cosmic Radiation↗

Neutrophil-endothelial cell interaction in critical illness.

Sepsis syndrome frequently results in endothelial injury in many organ systems. To evaluate neutrophil-pulmonary endothelial cell interaction in the sepsis syndrome, we studied 39 critically ill patients prospectively and 20 normal volunteers. Thirteen patients with sepsis (mean age, 71.4 years), 14 patients in an intensive care unit control group (mean age 65.4 years), and 12 patients admitted with acute myocardial infarction (mean age, 66.8 years) were evaluated. Blood samples were drawn from septic patients within 24 hours and from ICU and MI patients within 72 hours of admission. All sepsis patients were culture positive, 6 of 13 from the blood. Both renal failure and ARDS developed in 54 percent of septic patients. 51Cr-labelled neutrophils were prepared and added to bovine pulmonary endothelial cell monolayers with and without added phorbol myristate acetate. Endothelial cells with adherent PMA and nonadherent PMN's, were harvested and radioactivity in each fraction measured with a gamma scintillation counter. Baseline and maximally stimulated (PMA, 3.0 ng/ml) neutrophil adherence to endothelial cells were similar in all patients groups. However, in septic patients, PMA-stimulated PMN adherence was reduced at lower doses, most significantly in those who developed ARDS within 24 to 48 hours of admission (p less than 0.05). Seventy-one percent of patients who developed ARDS had reduced stimulated adherence (PMA 1.0 ng/ml) compared to 22 percent of critically ill patients who did not. We conclude that diminished adherence of neutrophils to endothelium in response to low-level PMA stimulation is significantly more common in patients with sepsis who develop ARDS. Our findings suggest that PMN-endothelial cell interaction is altered by the time sepsis is clinically recognized but before the development of ARDS. We speculate that the observed reduction in adherence of the PMN to endothelial cells may be a consequence of down-regulation by mediators generated in the inflammatory response to sepsis and/or the need for active participation of septic endothelium in this interaction.

Aged↗

Determination of CYP4A11-catalyzed lauric acid 12-hydroxylation by high-performance liquid chromatography with radiometric detection.

Lauric acid serves as an endogenous substrate for the cytochrome P450 enzyme CYP4A11. A reverse-phase, high-performance liquid chromatography method is described for the quantification of 12-hydroxylauric acid formed enzymatically by incubation of 14C-labeled lauric acid with cDNA-expressed CYP4A11 or human liver microsomes. Analytical separation is achieved using a C18 column and a gradient of 30% acetonitrile and 2 mM perchloric acid to 100% methanol, using a detection scintillation counter. This method is applicable to enzymatic studies for determination of lauric acid 12-hydroxylation activity.

Catalysis↗

Luciferases as reporter genes.

Luciferase is the ideal reporter gene to provide temporal and spatial information on promoter activity in Arabidopsis and other eukaryotes; the noninvasive detection of luminescence and short half-life of luciferase activity allow repeated measurements of individual seedlings over several days to assay dynamic changes in gene expression. Transgenic or transiently transformed plants with a luciferase gene under the control of a promoter of interest are required. Detection of the low level of luminescence produced by the luciferase gene in Arabidopsis requires the use of low light detecting charge-coupled device (CCD) cameras or scintillation counters. This chapter contains protocols on assaying and imaging luciferase in vivo and the automation for high-resolution timecourses.

Arabidopsis↗

[Solid phase radioimmunoassay for plasma testosterone using plastic microtiter tray (author's transl)].

UNLABELLED: In order to simply radioimmunoassay for plasma testosterone and to measure many samples at the same time, a method of solid phase radioimmunoassay utilizing a plastic disposable microtiter tray (DMT) by which chromatography can be omitted was investigated. METHOD: The antiserum was obtained by immunizing rabbits with testosterone-3 BSA which had been synthesized according to the Erlarnger's method. Plasma samples (male: 0.05ml, female: 0.2 ml) were extracted with 1.0 ml of ether. After freezing the plasma layer in an acetone-dryice bath, the ether phase was transfered to a glass tube and evaporated to dryness. These samples and the dried standard testosterone were dissolved with borate buffer containing 3H-testosterone and transfered to plastic DMT which had been precoated with the diluted antiserum, and incubated for 24 hrs. After removal of the incubated solution, the cups of DMT were cut off and were dissolved with toluene scintillator in counting vials. The radioactivity was counted with a liquid scintillation counter. RESULT: Other steroids except for 5alpha-dihydrotestosterone (5alpha-DHT) had a low degree of cross reactivity with the antiserum. Five alpha-DHT which could be measured together with testosterone in this assay was not a problem clinically because of its strong androgenic activity. The best standard curve was obtained when the antiserum was diluted to 1:1000. The sensitivity of this assay was 10 pg/tube. The maximal adsorption of antibody to plastic DMT was observed when the pH of antiserum was within the range of 6.5-9.5 and the precoating time was 24 hr at room termperature. The best pH of incubation buffer was 8.0, and the antigen-antibody reaction became a plateau when the incubation exceeded 6 hrs. Water blank in this assay was 4.6 +/- 2.1 pg/tube. The recovery of testosterone (50, 100, 200 pg) added to 0.1 ml female plasma was 99 +/- 6.8%. Coefficients of variation within assay and between assay were below 11.2% and 20.0%, respectively. Correlation between this method and the dextran-coated charcoal method was fairly good (r=0.938). Plasma testosterone levels in 10 normal males and 12 normal females were 616 +/- 202 (mean +/- SD) ng/dl and 66 +/- 29 (mean +/- SD) ng/dl, respectively. The levels were low in patients with hypopituitarism, hypogonadism and acromegaly. They were normal in patients with Cushing's syndrome due to adrenal hyperplasia and adenoma, but they were high in a patient with adrenal carcinoma. In a patient with testicular feminization, the level was 632 ng/dl. This increased after the administration of HCG, and decreased to 127.5 ng/dl after castration. This solid phase radioimmunoassay (using plastic DMT) is economically feasible as well as simple because it is possible to separate the bound hormone from the free hormone of all samples at the same time and there is little restriction in time and temperature. According to the above results, this method is suitable for routine clinical use.

Acromegaly↗

[A simple method measuring plasma aldosterone by radioimmunoassay without extraction (author's transl)].

A radioimmunoassay method was developed to measure plasma aldosterone levels. Antibody was produced in rabbits by injecting aldosterone oxime coupled with bovine gamma-globulin once a month. Plasma aldosterone was measured simultaneously by two methods: the direct method without extraction and a method using paper chromatography. 125I-aldosterone was used in the first method and 3H-aldosterone in the second. The antibody had a high specificity adequate to show zero water blank in the first method. Adequate precision, accuracy and sensitivity were obtained in a direct method using 125I-aldosterone. Plasma aldosterone levels were 7.1+/-3.0ng/100ml (Mean +/-SD) in normal subjects and slightly higher after injecting ACTH-Z. The correlative coefficient between the first and the second method was significantly high (r=0.970, P less than 0.001, n=37). Plasma aldosterone was high (34.3+/-14.1ng/100ml, n=7) in primary aldosteronism, slightly high (14.2+/-2.6ng/100ml) in secondary aldosteronism, normal in Cushing's syndrome (10ng/100ml) and low in Addison's disease (1ng/100ml), hypopituitarism (1ng/100ml) and pseudoaldosteronism (2ng/100ml). From these results, it is concluded that the direct method without extraction was a very useful and reliable method for measuring plasma aldosterone. It was superior in simplicity and there is no need to use a liquid scintillation counter.

Addison Disease↗

[A simple method for measuring aldosterone secretion rate using 125I-aldosterone (authors' transl)].

A simplified radioimmunoassay system for aldosterone secretion rate was developed by using radioiodine-labelled aldosterone and highly specific antiserum to aldosterone. An antibody was produced in rabbits by injecting aldosterone-oxime coupled with bovine gamma-globulin once a month. Aldosterone-oxime was labelled with 125I by using the chloramine T method described by Hunter and Greenwood. 3 ml of a twenty-four-hour urine specimen was used for the radioimmunoassay. Following CH2Cl2 extraction, pH 1 hydrolysis was carried out for twenty-four hours. Separation of the aldosterone extract was achieved by paper chromatography (hexane:benzene:methanol:water = 1:9:5:2.5). The minimum measurable concentration was under 1pg, and adequate intraassay and interassay precision were obtained. Aldosterone secretion rate was 89.6 +/- 25.8 (mean +/- SD)mug/day in normal subjects and was similar in low- and normal-renin essential hypertension. Significant high values (806.4 +/- 65.8mug/day) were obtained in primary aldosteronism and were slightly high in secondary aldosteronism and high-renin essential hypertension. The aldosterone secretion rate correlated positively with plasma aldosterone level (r = 0.731, p < 0.01). Aldosterone secretion rates were obviously higher than plasma levels in primary and idiopathic aldosteronism. From these results, it is concluded that this method is a very useful and reliable one for measuring aldosterone secretion rate and for discriminating primary aldosteronism from low-renin essential hypertension. it is superior in its simplicity, and there is no need to use a liquid scintillation counter.

Aldosterone↗

Radioenzymatic determination of epinephrine, norepinephrine and dopamine in 0.1 ml plasma samples: plasma catecholamine response to submaximal and near maximal exercise.

A sensitive enzymatic isotope derivative method for simultaneous determination of epinephrine, norepinephrine, and dopamine in 0.1 ml plasma samples is described. The assay consists basically of 1. conversion of epinephrine, norepinephrine, and dopamine into their respective methyl-derivates in the presence of catechol-O-methyltransferase and S-adenosylmethionine-[3H]methyl, 2. extraction of the methylated 3H labelled amines with diethyl ether, 3. separation by thin-layer chromatography, 4. measurement in a beta radiation scintillation counter. The effect of standing upright and of submaximal and near-maximal steady state exercise on plasma epinephrine, norepinephrine, dopamine and plasma renin was studied in 16 young adults aged 22 to 34 years and 9 physically well trained boys 16-17 years old. Standing upright for 2 min did not results in significant changes in either epinephrine, norepinephrine, dopamine or plasma renin activity compared with supine (basal) values. During exercise, epinephrine, norepinephrine, dopamine and plasma renin activity increased exponentially with the work load, peak values being attained at the highest load or immediately after cessation of exercise. The respective maximal values for plasma renin activity and epinephrine and norepinephrine levels were approximately 3, 6 and 10 fold higher than basal values in the adults, and rose as high as 4, 10 and 20 fold in the boys. Peak dopamine values were only 2-3 times higher than the basal values in both adults and boys. It is concluded that the method described provides a suitable tool for a quantitative assessment of adreno-sympathetic tone during physiological and pathophysiological conditions.

Adolescent↗

A radio-immunoassay of 19-nortestosterone using Celite column chromatography.

In order to measure plasma or serum concentrations of 19-nortestosterone (nandrolone), a radio-immunoassay has been developed using C7-tritium labelled nandrolone and antisera raised against nandrolone 3-(O-carboxymethyl)oxime-albumin. Because these antisera displayed considerable cross-reactions towards testosterone, 5 alpha-dihydrotestosterone and 5 alpha-dihydronandrolone, it proved to be necessary to separate nandrolone from these cross-reacting steroids prior to the radio-immunoassay. Using Celite/ethylene glycol columns a complete separation could be achieved between nandrolone and these cross-reacting steroids. The assay procedure involved extraction with ethyl acetate, chromatography on Celite/ethylene glycol columns, overnight incubation of samples and standards with antiserum and labelled compound, bound/free separation with dextran-coated charcoal and counting of the supernatant in a liquid scintillation counter. Recoveries for the total assay procedure (expressed as fractions) were between 0.85 and 1.01. The assay displayed a sensitivity of 0.7 nmol/l (at 50% binding) or 0.2 nmol/l (at 80% binding), while the within-assay and from day to day coefficients of variation were 9.8% (n = 114) and 12.9% (n = 13) respectively for duplicate measurements.

Animals↗

Retinol-induced changes in the phosphorylation levels of histones and high mobility group proteins from Sertoli cells.

Chromatin proteins play a role in the organization and functions of DNA. Covalent modifications of nuclear proteins modulate their interactions with DNA sequences and are probably one of the multiple factors involved in the process of switch on/off transcriptionally active regions of DNA. Histones and high mobility group proteins (HMG) are subject to many covalent modifications that may modulate their capacity to bind to DNA. We investigated the changes induced in the phosphorylation pattern of cultured Wistar rat Sertoli cell histones and high mobility group protein subfamilies exposed to 7 microM retinol for up to 48 h. In each experiment, 6 h before the end of the retinol treatment each culture flask received 370 KBq/ml [32P]-phosphate. The histone and HMGs were isolated as previously described [Moreira et al. Medical Science Research (1994) 22: 783-784]. The total protein obtained by either method was quantified and electrophoresed as described by Spiker [Analytical Biochemistry (1980) 108: 263-265]. The gels were stained with Coomassie brilliant blue R-250 and the stained bands were cut and dissolved in 0.5 ml 30% H2O2 at 60oC for 12 h. The vials were chilled and 5.0 ml scintillation liquid was added. The radioactivity in each vial was determined with a liquid scintillation counter. Retinol treatment significantly changed the pattern of each subfamily of histone and high mobility group proteins.

Animals↗

Atrazine biodegradation to deisopropylatrazine and deethylatrazine in coastal sediments of different land uses.

Atrazine, a triazine herbicide widely used in the United States, contributes to surface-water and groundwater contamination, as can deisopropylatrazine (DIA) and deethylatrazine (DEA), two of its microbial degradation products. Production of DIA and DEA by native bacteria in aquatic sediments has not been investigated thoroughly. We assessed atrazine and production of DIA and DEA over time in coastal aquatic sediments associated with different land uses including creeks from an undeveloped preserve and a suburban development, a golf course drainage ditch, and a contaminated commercial harbor. Sediments were incubated in microcosms, spiked with U-14C-atrazine, extracted, and analyzed for 14C in a liquid scintillation counter. Atrazine, DIA, and DEA also were quantified by gas chromatography-mass spectrometry. The amount of 14C recovered varied at each site as a function of the sediment organic carbon content and decreased significantly over time. Both DEA and DIA were measured primarily in the aqueous phase. Transformation was more extensive to DIA than to DEA. The ratio of DIA to atrazine recovered from the undeveloped preserve was as high as 0.13. In contrast, the golf course had limited biotransformation, and had the greatest atrazine recoveries so atrazine, not DEA and DIA, may have a greater impact at this site.

Atrazine↗

Induction of unscheduled DNA synthesis in hairless mouse epidermis by skin carcinogens.

Induction of unscheduled DNA synthesis (UDS) in hairless mouse epidermis by six chemicals was determined in an in vivo-in vitro assay by using a liquid scintillation counting method. Test chemicals were applied once onto two areas of the back of female hairless mice after stripping of the stratum corneum with adhesive tape to enhance skin penetration. After exposure, the skin samples were taken and cultured in a medium containing [3H]thymidine with or without hydroxyurea (HU, an inhibitor of replicative DNA synthesis). DNA of the epidermis was extracted, and incorporation of [3H]thymidine into DNA and the DNA content was determined with a liquid scintillation counter and a fluorescence spectrophotometer, respectively. Induction of UDS by chemicals was judged by calculation of the UDS index [(the ratio of DNA synthesis in the presence of HU to that in its absence) x 100]. A good correlation between UDS induction and organ specificity of carcinogens was observed. 4-Nitroquinoline 1-oxide, a skin carcinogen used as a positive control, induced a dose-dependent increase in the UDS index of approximately 12-fold at 2 hr after exposure, while 1,2-epoxydodecane, a non-skin carcinogen applied as a negative control, did not increase the UDS index. Four other skin carcinogens induced dose-dependent increases in the UDS index; N-methyl-N'-nitro-N-nitrosoguanidine and diepoxybutane at 2 hr after exposure, and 7,12-dimethylbenz[a]anthracene and benzo[a]pyrene at 24 hr after exposure. The results suggest that UDS is a good marker of the genotoxicity of skin carcinogens.

4-Nitroquinoline-1-oxide↗

Induction of unscheduled DNA synthesis in hairless mouse epidermis by ultraviolet light.

The activity of ultraviolet (UV) light to induce unscheduled DNA synthesis (UDS) was investigated in hairless mouse epidermis by means of an in vivo-in vitro assay using a liquid scintillation counting method. Groups of three to five 8-week-old female hairless mice were irradiated with UV-B or UV-A, then skin samples were taken and cultured individually in medium containing [3H]thymidine with or without hydroxyurea (HU) for 2 hr. DNA of the epidermis was extracted, and incorporation of [3H]thymidine and the DNA content were determined with a liquid scintillation counter and a fluorescence spectrophotometer, respectively. Induction of UDS was judged in terms of the UDS index [(the ratio of DNA synthesis in the presence of HU to that in its absence) x 100]. UV-B increased the UDS index 1 hr after irradiation of 500 J/m2, which corresponds to approximately 1 minimal erythema dose or 1 minimal edema dose, and showed a dose-dependent increase up to 17-fold in the UDS index at irradiation doses of 500 to 2,000 J/m2. In a time-course study, UV-B also increased replicative DNA synthesis (RDS) 48 hr after irradiation at 1,000 J/m2. On the other hand, UV-A did not increase the UDS index at irradiation doses of 2 x 10(5) to 8 x 10(5) J/m2. These results show that induction of UDS by UV irradiation depends on wavelength and an increase of RDS in the epidermis exposed to UV-B irradiation appears after induction of UDS.

Animals↗

Effect of aging on unscheduled DNA synthesis induction by UV-B irradiation in hairless mouse epidermis.

Age-related induction of unscheduled DNA synthesis (UDS) by ultraviolet B (UV-B) irradiation was investigated in the epidermis of female hairless mice by means of an in vivo--in vitro assay using a liquid scintillation counting method. Skin samples were taken and cultured in a medium containing [3H]thymidine with or without hydroxyurea (HU) for 2 hr. DNA of the epidermis was extracted, and the incorporation of [3H]thymidine into DNA and the DNA content were determined with a liquid scintillation counter and a fluorescence spectrophotometer, respectively. Induction of UDS by UV-B was judged in terms of the value of the UDS index calculated as a percentage of the respective unexposed control value taken as 100% [the UDS index is given by (the ratio of DNA synthesis in the presence of HU to that in its absence) x 100]. DNA synthesis both in the presence and absence of HU decreased with age [12 months old (M) < 8 M < 4 M < 2 M < 1 M)], concomitantly with a small but significant increase of UDS index. The decrease was high in the younger age groups and moderate in the older age groups. UV-B increased the UDS index approximately 14-, 12- and 9-fold at 1 hr after 1,000 J/m2 irradiation in 1 M, 2 M and 12 M mice, respectively, and these increases were partly reversed at 4 hr after irradiation. UV-B also increased the UDS index approximately 25-, 24- and 21-fold at 1 hr after 4,000 J/m2 irradiation for each age group. However, there was no statistically significant age-related difference in the magnitude of the UDS index after irradiation of UV-B. These results show that replicative DNA synthesis decreases with age, whereas DNA repair capacity after UV-B irradiation does not change with age under these conditions.

Aging↗

Induction of unscheduled DNA synthesis in hairless mouse epidermis by 8-methoxypsoralen plus ultraviolet A (PUVA).

Induction of unscheduled DNA synthesis (UDS) by 8-methoxypsoralen (8-MOP) plus ultraviolet A (UV-A) (PUVA) was investigated in the epidermis of female hairless mice by means of an in vivo--in vitro assay using a liquid scintillation counting method. Groups of three to five 8-week-old female hairless mice had 8-MOP applied once onto two areas of the back after stripping of the stratum corneum with adhesive tape to enhance skin penetration, and were irradiated with UV-A. Skin samples were taken and cultured in a medium containing [3H]thymidine with or without hydroxyurea (HU) for 2 hr. DNA of the epidermis was extracted, and the incorporation of [3H]thymidine into DNA and the DNA content were determined with a liquid scintillation counter and a fluorescence spectrophotometer, respectively. Induction of UDS was judged in terms of the UDS index [(the ratio of DNA synthesis in the presence of HU to that in its absence) x 100]. In a time-course study, the UDS index was increased at 1, 2 and 24 hr after 1 x 10(5) J/m2 UV-A irradiation with 0.001% 8-MOP, reaching the maximum level at 24 hr. In a dose-response study, it was significantly increased at the dose of 1 x 10(5) J/m2 of UV-A at 24 hr with 0.001% 8-MOP, but showed no significant change at the doses of 0.5 x 10(5), 2 x 10(5) and 4 x 10(5) J/m2. In a further study on the effect of varying the dose of 8-MOP, the UDS index was significantly increased at 0.001 and 0.002% 8-MOP at 24 hr after 1 x 10(5) J/m2 UV-A irradiation, reaching the maximum level with 0.002% 8-MOP. The increase of the UDS index in these studies was less than 3-fold. These results show that PUVA causes a small induction of UDS, which might be due to slow DNA excision repair over a long period.

Animals↗

A multi-modality assay platform for ultra-high throughput screening.

The demand for increased throughput during primary screening using less reagents is changing the way of drug discovery. Searching for hits using high throughput screening in 96-well format plates is being replaced by the use of higher density plates, such as 384 and 1536-well formats. The analysis of radiometric assays by scintillation counters is becoming limiting since only 12 wells can be counted at a time. Charged coupled device (CCD) camera based instruments, that image the whole plate in one exposure, speed up detection and are compatible with any microplate footprint. Researchers are also demanding a choice of detection methods, including fluorescence, luminescence and radioactivity, and require imagers suitable for all applications. LEADseeker Homogenous Imaging System is a multi-modality platform offering imaging technology and assay toolboxes for radiometric, fluorescent and luminescent based assays. LEADseeker allows the very rapid analysis of high density formats enabling ultra-high throughput screening of a range of biological assays. Research areas that can be studied using this system include enzyme assays, receptor binding and molecular interactions.

Drug Design↗