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Biomedical subjects

A Johnsson

Publications and source records attributed to A Johnsson.

At least 19 recordsLinked to original sources

Identification of gene clusters differentially expressed during the cellular injury responses (CIR) to cisplatin.

The goal of this study was to identify changes in mRNA levels in tumour cells after a toxic exposure to cisplatin (IC(99)dose). Using suppression-subtractive hybridization (SSH) 2 cDNA libraries were created, an UP library (202 cDNA fragments) and a DOWN library (153 cDNA fragments). Using reversed Northern hybridization 16 and 30 fragments were truly differentially expressed in the UP and DOWN libraries, respectively. Most prominent in the UP library were the mitochondrial and injury response clusters and in the DOWN library the cytoskeletal, protein synthesis and signalling clusters. These distinct clusters potentially represent an expression profile of the cisplatin-induced cellular injury response.

Antineoplastic Agents↗

Antitumor effect of radioactive cisplatin (191Pt) on nude mice.

PURPOSE: To investigate the effect of (191)Pt-cisplatin in vivo in terms of the antitumor effect and general toxicity on tumor-bearing nude mice. METHODS AND MATERIALS: Tumor-bearing (human squamous cell carcinoma, AB) nude mice were divided into four groups and given, i.p., physiological saline (controls), cisplatin, (191)Pt-cisplatin (80 MBq/mg), or (191)Pt-cisplatin (160 MBq/mg), respectively. Mortality and weight were used as parameters for monitoring general toxic effect, while specific growth delay (SGD) and the area under the logarithm of the relative tumor size curve (AUC-log[RTS]) were used to evaluate the antitumor effect of the treatments. RESULTS: Both SGD and AUC-log(RTS) values showed that (191)Pt-cisplatin was significantly (P < 0.05) more effective in retarding tumor growth than nonradioactive cisplatin. No differences in mortality between the different groups could be observed and no significant differences in weight change between the mice treated with cisplatin or (191)Pt-cisplatin could be seen. CONCLUSION: (191)Pt-cisplatin is a more effective drug than nonradioactive cisplatin in retarding tumor growth on nude mice without adding systemic toxic effects. We believe that radioactive cisplatin may prove to be an alternative to conventional cisplatin; however, the possible toxic effects on organs at risk have to be thoroughly investigated.

Animals↗

Autonomic straightening of gravitropically curved cress roots in microgravity.

The typical response of plant organs to gravistimulation is differential growth that leads to organ bending. If the gravitropic stimulus is withdrawn, endogenous compensation of the graviresponse and subsequent straightening occur in some plants. For instance, autonomic straightening of Lepidium roots occurs when gravitropically-curved rootsare rotated on a clinostat (Stankovi et al., 1998a). To determine whether endogenous compensation of the graviresponse also occurs in space, microgravity-grown cress roots were laterally centrifuged in-flight and then returned to microgravity using Biorack hardware on a shuttle mission (STS-81). The cress roots were centrifuged at 4 different g-doses (0.1 x g and 1 x g for 15 or 75 min). All four treatments yielded varying degrees of root curvature. Upon removal from the centrifuge, roots in all four treatments underwent subsequent straightening in microgravity. This straightening resulted from a loss of gravitropic curvature in older regions of the root and the coordinated alignment of new growth. These results show that both microgravity and clinostat rotation on Earth are equivalent in stimulus withdrawal with respect to the induction of endogenous compensation of the curvature. Cress roots are the only plant organ shown to undergo compensation of the curvature in both microgravity and on a clinostat. The compensation of graviresponse in space rules out the hypothesis that the endogenous root straightening ("autotropism") represents a commitment to a pre-stimulus orientation with respect to gravity and instead suggests that there is a default tendency towards axiality following a withdrawal of a g-stimulus.

Centrifugation↗

In vitro toxicity of (191)Pt-labeled cisplatin to a human cervical carcinoma cell line (ME-180).

PURPOSE: The aim of the present work was to examine the effect of (191)Pt-cisplatin, and to study the manner in which radiation and cisplatin interact, in a human cervical carcinoma cell line (ME-180). METHODS AND MATERIALS: The cells were incubated for 1 hour with nonradioactive cisplatin or (191)Pt-cisplatin with specific activities in the range 48-167 MBq/mg. The surviving fraction of the cells after 7 days' growth was determined with a nonclonogenic tetrazolium-based (MTT) assay. The uptake of platinum into the cell and the amount of platinum bound to DNA was measured. RESULTS: The 50% inhibition concentration (IC(50)) decreased with increasing specific activity of the (191)Pt-cisplatin. For the specific activities 0 (nonradioactive), 48, 89, 143, 157, and 167 MBq/mg, IC(50) was found to be 3.24 +/- 0.08, 2.77 +/- 0.55, 2.17 +/- 0.34, 1.15 +/- 0.04, 1.02 +/- 0.03, and 0.76 +/- 0.13 respectively. Isobologram analysis showed a supra-additive (synergistic) interaction between the radiotoxicity and chemotoxicity for specific activities over 100 MBq/mg. CONCLUSION: The cytotoxic effect of cisplatin may be enhanced by labeling the drug with the radionuclide (191)Pt.

Antineoplastic Agents↗

An open study of efficacy and safety of long-term treatment with mometasone furoate fatty cream in the treatment of adult patients with atopic dermatitis.

BACKGROUND: Atopic dermatitis is a severe chronic skin disease often deteriorated by the presence of microorganisms and often responds well to treatment with potent corticosteroids. However, the long-term use of potent topical corticosteroids are accompanied by side-effects such as skin atrophy. OBJECTIVE: To study the effect and safety of prophylactic treatment with mometasone furoate fatty cream (contains hexylene glycol) for 6 months in patients with atopic dermatitis. RESULTS: Sixty-one of 68 (90%) patients were still free of their disease after 6 months of twice weekly treatment and only one showed possible treatment related signs of skin atrophy. The number of Staphylococcus aureus and Pityrosporum ovale were significantly reduced in cleared patients. CONCLUSIONS: Mometasone furoate fatty cream is effective and safe both for treatment and as a prophylaxis in patients with atopic dermatitis.

Administration, Topical↗

Identification of genes differentially expressed in association with acquired cisplatin resistance.

The goal of this study was to identify genes whose mRNA levels are differentially expressed in human cells with acquired cisplatin (cDDP) resistance. Using the parental UMSCC10b head and neck carcinoma cell line and the 5.9-fold cDDP-resistant subline, UMSCC10b/Pt-S15, two suppressive subtraction hybridization (SSH) cDNA libraries were prepared. One library represented mRNAs whose levels were increased in the cDDP resistant variant (the UP library), the other one represented mRNAs whose levels were decreased in the resistant cells (the DOWN library). Arrays constructed with inserts recovered from these libraries were hybridized with SSH products to identify truly differentially expressed elements. A total of 51 cDNA fragments present in the UP library and 16 in the DOWN library met the criteria established for differential expression. The sequences of 87% of these cDNA fragments were identified in Genbank. Among the mRNAs in the UP library that were frequently isolated and that showed high levels of differential expression were cytochrome oxidase I, ribosomal protein 28S, elongation factor 1alpha, alpha-enolase, stathmin, and HSP70. The approach taken in this study permitted identification of many genes never before linked to the cDDP-resistant phenotype.

Cisplatin↗

Effects of 50 Hz electric currents and magnetic fields on the prokaryote Propionibacterium acnes.

The effects of 50 Hz sinusoidal electric currents and magnetic fields on the Gram-positive skin bacterium Propionibacterium acnes were investigated. Intracellular free calcium ([Ca(2+)](i)), intracellular pH (pH(i)), and cell viability were examined, based on their relevance to ELF field studies and on previous studies conducted on P. acnes (UVA irradiation, photosensitization using porphyrin-based sensitizers, and broad-band red light). The [Ca(2+)](i) and the pH(i) were measured spectrofluorimetrically using the fluorescent probes fura-2 and BCECF, respectively. Sham-exposed controls were used to assess the field exposed samples. Cell suspensions were exposed to 50 Hz, 0.2 mT sinusoidal magnetic fields generated by using Helmholtz coils for up to 30 min. The estimated maximum induced electric field was 0.2 mV/m. Changes in [Ca(2+)](i) and cell viability were not detected. Ag/AgCl electrodes were used to expose cell suspensions to 50 Hz sinusoidal electric currents. The current densities were in the range 0.015-1500 A/m(2) (corresponding electric fields congruent with0.01-1000 V/m). Changes in [Ca(2+)](i) were not observed after current exposure. Current densities of 800 A/m(2) (electric field E congruent with550 V/m) were required for a 50% reduction in cell viability. Current densities greater than 800 A/m(2) were required for a reduction in pH(i). However, a pH gradient across the cell membrane (inside alkaline) was maintained even when exposure resulted in less than 0. 2% survival (1400 A/m(2), E congruent with950 V/m). Thus, dissipation of the pH gradient across the cell membrane and changes in [Ca(2+)](i) were not a consequence of cell inactivation by 50 Hz electric currents. This is in contrast to inactivation of P. acnes by UVA irradiation or photosensitization, where such changes have been obtained.

Calcium↗

Expression of GADD153 in tumor cells and stromal cells from xenografted tumors in nude mice treated with cisplatin: correlations with cisplatin-DNA adducts.

PURPOSE: Cisplatin is a commonly used antineoplastic agent that acts by forming adducts with DNA, and causing a response to the cellular injury. One of the components of this cellular injury response is the activation of the "growth arrest and DNA damage gene" GADD153. The level of GADD153 induction in tumor cells has been associated with the degree of cytotoxicity. The pupose of this study was to determine whether cisplatin activates GADD153 also in nontumor cells and how GADD153 protein levels correlate with cisplatin-DNA adducts in different cell types. METHODS: Nude mice with xenografted squamous cell carcinoma were treated with cisplatin 10 mg/kg. Tumors were removed at 0 h (untreated controls), 24 h, and 48 h and immunohistochemically stained for GADD153 protein and cisplatin-DNA adducts. The staining reaction was quantitated in tumor cells and nonmalignant stromal cells separately, using computerized image analysis. RESULTS: The GADD153 level was 4.5 times higher in tumor cells than in stromal cells in untreated mice. At 24 h after cisplatin treatment the GADD153 level had increased by 50% and 72% in tumor cells and stromal cells, respectively. Analysis of the cisplatin-DNA adducts showed a reversed pattern, with six-fold higher levels in stromal cells than in tumor cells at 24 h after treatment. By combining these data, we estimated that approximately 25-fold more GADD153 per cisplatin-DNA adduct was induced in tumor cells than in stromal cells. CONCLUSION: Our data suggest that different cell types may respond differently to DNA damage caused by cisplatin.

Animals↗

Effects of amifostine on cisplatin induced DNA adduct formation and toxicity in malignant glioma and normal tissues in rat.

The chemoprotective effect of amifostine (WR2721) was studied in a BDIX rat model with intracerebral BT4C glioma implants. Twenty-one rats were given cisplatin 5 mg/kg i.p., 21 were given amifostine 200 mg/kg i.p. + cisplatin 5 mg/kg i.p. Ten rats served as untreated controls. An immunohistochemical method for analysis of cisplatin-DNA adducts was used to elucidate the adduct formation in tumor, normal brain and kidney. Tumor volume and serum creatinine level were analysed 10 days after treatment. In animals pretreated with amifostine there was a delayed adduct formation rate in the normal brain, and in the kidney cortex the number of tubular cells with extremely high adduct level was reduced. No difference in adduct formation was seen in tumors. Tumor volume was significantly larger following amifostine + cisplatin (66% of controls) compared to cisplatin alone (38% of controls). Weight loss was, however, severe in rats given cisplatin alone. In the tumor growth study only 3 out of 11 rats treated with cisplatin 5 mg/kg alone survived until time of sacrifice at 10 days, whereas all those pretreated with amifostine survived. Mean serum creatinine was 48 micromol/l (controls), 146 micromol/l (cisplatin) and 59 micromol/l (amifostine + cisplatin). A marked reduction of histopathological renal changes was found when amifostine was added. Amifostine thus significantly reduced general and renal toxicity of cisplatin. The tumor growth retardation was stronger when cisplatin was given alone but this is probably related to general toxicity and malnutrition indirectly supported by the fact that amifostine did not significantly reduce cisplatin-DNA adduct formation in tumors. The results of the present study suggest that amifostine may have a role in increasing the therapeutic ratio of cisplatin, also in the treatment of malignant glioma.

Amifostine↗

Nuclear magnetic resonance imaging in studies of gravitropism in soil mixtures.

Gravitropic responses of oat coleoptiles were measured in different growth media; humid air, natural soil and artificial soil (glass beads). The oat coleoptiles in soil and glass beads were monitored by NMR imaging, while those in humid air were imaged in darkness with an infrared-sensitive charge-coupled device (CCD) camera. The present study shows for the first time that gravitropic experiments can be performed in artificial soil using NMR imaging as a convenient and suitable recording method. Not only was it possible to follow the gravitropic curvatures in natural soil, but the artificial soil allowed plant images of sufficient spatial and temporal resolution to be recorded. The advantages of using artificial soil in magnetic resonance imaging studies are that the iron content of glass beads is very low compared with natural soil, and that the artificial soil matrix can easily, be standardized with regard to particle size distribution and nutrient content. Two types of glass beads were used, the diameter of the small and the large beads being 300-400 and 420-840 micrometers, respectively. The growth rate of the coleoptiles in soil and in big beads was roughly the same and only slightly lower than in humid air, whereas small beads reduced the growth rate by approx. 16%. The bending rate of the coleoptiles during the gravitropic response was reduced by c. 65% in soil and 75% in bead mixtures relative to bending in air. It should be noted, however, that the maximum curvature of the coleoptile tip was of the same order in all cases, about 35 degrees. This value may represent the largest possible curvature of the organ. The potential of NMR imaging to study, how plant organs penetrate the soil under the influence of gravitropism, mechanical impedance and thigmotropism is also discussed.

Avena↗

Amifostine as a protector against cisplatin-induced toxicity in nude mice.

The mechanisms mediating the protective effects of amifostine on cisplatin-induced toxicity were investigated in tumor-bearing nude mice by quantitative immunohistochemistry for analysis of cisplatin-DNA adduct levels in tumors and kidneys. The mice were treated with cisplatin 5 or 10 mg/kg i.p. with or without amifostine 200 mg/kg 30 min prior to cisplatin. Toxicity was noted in terms of mortality and changes in body weight. Mortality was similar in the four treatment groups, regardless of cisplatin dose or whether amifostine was added or not. At a cisplatin dose of 5 mg/kg, amifostine did not affect the moderate decrease in body weight. Cisplatin 10 mg/kg alone gave a significant loss of body weight, with the nadir on day 7. By adding amifostine to 10 mg/kg cisplatin the weight loss was much less pronounced. Tumor growth was significantly more retarded among animals treated with 10 mg/kg cisplatin alone compared with amifostine + cisplatin 10 mg/kg. There was no difference in tumor growth retardation between cisplatin 5 mg/kg alone or in combination with amifostine. The most likely explanation was that the pronounced tumor growth retardation with 10 mg/kg cisplatin alone was due to the decline in the general condition of the animals rather than increased antitumoral activity per se. Analysis of cisplatin-DNA adducts in tumors showed no difference whether cisplatin 10 mg/kg was combined with amifostine or not. In kidneys there were significantly fewer tubular cells with very high adduct levels in animals pretreated with amifostine.

Amifostine↗

Circumnutations without gravity: a two-oscillator model.

Helical growth movements termed circumnutations occur in a number of different plant organs. The underlying mechanism are complicated. They consist of an endogenous as well as an erogenous component. Since circumnutations in sunflower hypocotyls occur also under gravity free conditions in Space, gravity is not necessary for them to arise. However, the changes of period and amplitude of circumnutations with the applied gravitational force indicate that gravity plays a modifying role. Models for circumnutations have to take these features into account. It seems natural to study minimum models with at least two oscillating units. Two-oscillator models for circumnutations have already been discussed. We have studied the interaction between two coupled oscillatory units. With this model we could simulate movements of hypocotyls. The results of simulations were compared with experimentally recorded movements of the Arabidopsis hypocotyl.

Arabidopsis↗

Sample handling for determination of free platinum in blood after cisplatin exposure.

Cisplatin is a commonly used cytostatic drug that can be pharmacokinetically monitored by measurement of non-protein-bound platinum (Pt) in plasma. The present report elucidates some practical aspects on blood sample preparation with the purpose of identifying methodological error sources and simplifying sample handling. Human blood was incubated in vitro with clinically relevant doses of cisplatin and then stored at different temperatures for various periods prior to deproteinization. Pt concentrations were analyzed by inductively coupled plasma mass spectrometry, which is a very sensitive technique that is well suited for determination of free Pt in biological samples. Free Pt concentration is usually determined after ultrafiltration of plasma. An alternative approach used to study the protein-free fraction is to precipitate the proteins with ice-cold ethanol and then analyze the aliquot. We compared the Pt concentrations in protein-free plasma obtained by these two methods and the levels were very similar. Thus, ethanol deproteinization may be an attractive alternative to ultrafiltration, since it is both simple to perform and very cost-effective. Storage of nondeproteinized whole blood or plasma at 4 degrees C or 20 degrees C did not affect the free Pt fraction significantly for the first 2 h, after which a gradual decrease was noted. This suggests that saving the blood sample for approximately 1 h before processing should not have any major impact on the result. On long-term storage of frozen blood samples prior to deproteinization the freezing temperature proved to be of critical importance. At -20 degrees C there was a gradual decrease in the free Pt fraction during the study period of 14 days, whereas the free Pt concentration remained unchanged at -70 degrees C for at least 3 months. Thus, samples may be stored unprocessed at -70 degrees C for several months, whereas -20 degrees C is not sufficiently cold.

Cisplatin↗

Effects of microgravity on the growth of Lepidium roots.

The normal growth dynamics of plant roots is partly controlled by the gravitational force. In order to study the detailed growth behavior in absence of gravity, the growth of Lepidium sativum roots was recorded by time lapse photography at 1h intervals in a Spacelab ESA-experiment (IML-2). Plants were germinated and kept in microgravity during the experiments, while control roots were at 1 g with normal static gravistimulation. Extended image analyses allowed new information to be achieved about movements of all parts of the roots, extending earlier published results. Root contours were extracted from the images and divided into 0.6mm segments. Deviation angles were calculated for each root segment, both for the first 8-10 h (phase I) and for the last 6-8 h of the experiment (phase II). For phase I, the present analysis confirmed that the average square deviation increased linearly with time for roots in microgravity, while for roots under 1 g conditions it stayed constant. This was consistent with a random walk hypothesis for the bending pattern. In phase II, roots in microgravity stopped their spontaneous curvatures and showed more straight growth or even diminished the root curvatures that had occurred during phase I. Thus, the growth is distinctly different in the two phases and is thought to be controlled by autotropic reactions in phase II. Root hairs developed when the roots passed into phase II. During phase I, the root growth rates were equal in microgravity and on the ground (0.50 mm h-1 with SE 0.04 and 0.51 mm h-1 with SE 0.03, respectively). In phase II the growth rate on the ground was higher than in microgravity (1.44 mm h-1 with SE 0.10 and 1.07 mm h-1 with SE 0.04 in microgravity). Microgravity conditions, therefore, clearly affect Lepidium root growth: In phase I the bending pattern is random in contrast to the normal straight growth under 1g. In phase II the growth rate is reduced, as compared to the growth rate under 1 g.

Brassicaceae↗

Circumnutations: results from recent experiments on Earth and in space.

Results of experiments on circumnutations from the last few decades are discussed, particularly in regard to the relationship between gravity and circumnutations. Pertinent and new results from experiments in microgravity as well as results from ground-based studies are summarized, and questions with relevance to space activities are posed. Theoretical approaches to describe aspects of circumnutation are also discussed. The review indicates some areas where further research efforts are needed.

Arabidopsis↗

Effects of cisplatin and amphotericin B on DNA adduct formation and toxicity in malignant glioma and normal tissues in rat.

In an attempt to modify the cytotoxicity of cisplatin, amphotericin B (AmB) was given as pretreatment to BDIX rats with intracerebral BT4C glioma implants. Ten animals given AmB 5 mg/kg i.p. followed by cisplatin 5 mg/kg i.p. displayed massive haematuria within 24 h after treatment and died a few days later. The antitumoral effect could not, therefore, be evaluated. Histopathological examination of the kidneys showed extensive tubular necrosis. No signs of apoptotic cell death were found using in situ end labelling with biotin-labelled nucleotides or with DNA integrity analysis in agarose gel electrophoresis. An immunohistochemical method for analysis of cisplatin-DNA adducts was used to elucidate the distribution of cisplatin in brain tumour, normal brain and kidney. Addition of AmB to cisplatin caused increased adduct formation in kidneys, particularly in tubular cells. It seems plausible that the nephrotoxicity, at least in part, was mediated by increased levels of cisplatin-DNA adducts. Pretreatment with AmB did not have any obvious effect on the formation of adducts in the cerebral cortex. The adduct levels in the tumours from animals pretreated with AmB were not significantly increased compared with those treated with cisplatin only. Thus, addition of AmB to cisplatin caused excessive nephrotoxicity suggesting a decrease in the therapeutic ratio of cisplatin.

Amphotericin B↗

Porphyrin sensitization and intracellular calcium changes in the prokaryote Propionibacterium acnes.

Photosensitization induces intracellular free calcium changes ([Ca2+]i) in some eukaryotic cell systems which either contribute to or protect against cell inactivation. We have investigated whether or not similar changes can be induced in prokaryotes. The skin bacterium Propionibacterium acnes was sensitized using protoporphyrin IX (PP IX) or 5-aminolevulinic acid (ALA). Exogenous ALA resulted in either a preferential accumulation of protoporphyrin (ALA-PP) or of coproporphyrin and/or uroporphyrin (ALA-CP/UP) in P. acnes. For PP IX or ALA-PP sensitization, exposure to broad-band red light resulted in an increase in [Ca2+]i. For ALA-PP sensitization, this increase was transient and [Ca2+]i returned to basal levels within 5-10 min after irradiation. However, the elevated [Ca2+]i levels obtained after PP IX sensitization were maintained for at least 1 h after irradiation. In both cases, the reduction in the external calcium concentration led to an enhancement in the cell survival, indicating that induced [Ca2+]i changes may participate in photoinactivation. Sensitization by hydrophilic coproporphyrin and/or uroporphyrin (ALA-CP/UP) did not affect the [Ca2+]i levels, but higher levels of cell inactivation were obtained. It therefore appears that damage to membrane-associated components is at least partly responsible for [Ca2+]i alterations after photosensitization.

Aminolevulinic Acid↗

Agravitropic behaviour of roots of rapeseed (Brassica napus L.) transformed by Agrobacterium rhizogenes.

Transgenic hairy roots of Brassica napus (cv. Omega) have been developed, using Agrobacterium rhizogenes strain AR 25, for use as a model system in the investigation of physiological and morphological differences between transgenic and normal roots. The basic parameters of growth and normal or altered gravitropical behaviour of hairy roots are for the first time presented in this paper together with an ultrastructural and morphological analysis of the root statocytes. The results obtained also represented the basis for the TRANSF0RM-experiment on the IML-2 mission performed onboard the Space Shuttle Columbia. Typical hairy root traits such as hormone-autonomous growth high growth rate, lateral branching, and changed/absence of gravitropism were detected. The transformed nature of the roots was confirmed by Southern blot analyses. The gravitropical behaviour of apices from hairy root cultures of this clone has been compared with root tips from normal seedlings. While the wild type roots curved progressively with increasing stimulation angles, the transformed roots showed no curvature when stimulated at 45 degrees, 90 degrees or 135 degrees on the ground. The morphology and ultrastructure of the root tip regions were examined by light microscopy and transmission electron microscopy. At the ultrastructural level no major differences could be detected between the roots studied. There was, however, a slight reduction in the starch content of most of the amyloplasts of the transgenic root tips, and the root cap was more V-shaped in the transgenic roots than in the wild type. Preliminary results from the Shuttle experiment TRANSFORM show a random distribution of amyloplasts in the root cells of both transformed and wild type root caps after 14 h on a 1xg centrifuge followed by 37 h in microgravity.

Agrobacterium tumefaciens↗