Exp Appl Acarol, 2004, 34(3-4), 335 - 44 Penetration of light into carpet, and responses of the European house-dust mite (Dermatophagoides pteronyssinus) to varying light intensity and diurnal cycle; Causer S et al.; This study examined the question of what effect exposure to light might play in determining the vertical distribution of house-dust mites in carpet, and the degree to which light penetrates worn and unworn carpets of different pile conformation (loop- versus cut-pile), height and colour . The effect on population increase of a diurnal lighting cycle versus continual darkness was also investigated . It was found that the penetration of light into carpets was largely unaffected by pile colour or conformation . Pile height was an important factor, however, and for a given height within the pile, light intensity was higher in carpets subjected to a greater degree of wear . This corresponded to the reduction in effective pile height that occurs with carpet use . Whilst the differences observed were sometimes large (up to two-fold for a given height within the pile), Petri dish studies suggested no mite preference for habitation of areas of low light intensity compared to high intensity . Additionally, culturing mites under a diurnal light cycle was shown to be no more efficient than culturing in complete darkness . These results suggest that exposure to light is not an important determinant of house-dust mite behaviour, or their ability to colonise textile substrates. Cytometry A . 2005 Jan 13; {Epub ahead of print} A novel approach to prepare extended DNA fibers in plants; Li L et al.; BACKGROUND: The extended DNA fiber preparation procedure is still imperfect in plants due to the existence of a hard cell wall; thus, high quality of extended DNA fibers for fluorescence in situ hybridization (FISH) analysis is often difficult to be obtained rapidly and efficiently . In this study we have developed a fast and widely effective method to prepare DNA fibers from various plant species and the fibers are suitable for fiber FISH mapping . METHODS: Fresh young leaves were chopped with a sharp sterile scalpel in a Petri dish that contained ice-cold nucleus isolation buffer followed by filtration through 33-mum nylon mesh . Nuclei were obtained by centrifuging the filtrates at high speed (16,000g) for 40 s . Nucleus lysis buffer (0.5% sodium dodecylsulfate, 5 mM ethylenediaminetetraacetic acid, 100 mM Tris, pH7.0) was added to nuclei on slides, and DNA fibers were dragged and extended with a clean coverslip . RESULTS: The key of this method is that liquid nitrogen grinding of leaves is replaced by chopping with a blade in ice-cold nucleus isolation buffer . With the liquid nitrogen method, over- or under-grinding of leaves occurs more frequently, and DNA fibers with the desired quality are not obtained easily . In contrast, it is easier to release nuclei from cells in nucleus isolation buffer by chopping, which results in fewer nuclei being destroyed . Highly extended, intact, and long DNA fibers can be generated to a great probability with this method . In addition, this method is very simple and rapid, requiring only 20 min for the entire process, and is also safe because poisonous mercaptoethanol is replaced by dithiothreitol . The results of fiber-FISH with maize genomic DNA and 45S rDNA as probes showed that DNA fiber size as long as 1.96 Mb could be measured . The successful and reliable preparation of maize, wild rice, and barley DNA fibers suitable for FISH mapping proves that this technique is a widely effective approach for obtaining extended DNA fibers in plants . CONCLUSIONS: A simple, rapid, safe, and widely effective method for getting extended DNA fibers has been developed in plants . (c) 2005 Wiley-Liss, Inc. Ann Bot (Lond) . 2005 Jan 11; {Epub ahead of print} Seed Germination and Seedling Emergence of Three Annuals Growing on Desert Sand Dunes in China; Tobe K et al.; * Background and Aims Information on the initial growth characteristics of annuals found in Chinese deserts is very limited . The aim of this study was to investigate seed germination and interactive effects of irrigation and seed burial depth in sand on seedling emergence and seedling survival in three annuals (Agriophyllum squarrosum, Bassia dasyphylla and Aristida adscensionis) commonly growing on sand dunes in these regions . * Methods Effects of temperature, light and polyethylene glycol-6000 on seed germination were examined by irrigating seeds sown on filter paper in Petri dishes . Seedling emergence was examined for seeds sown on the surface of, or at different depths (5, 10, 20, 30, 40 and 50 mm) in, sand-filled pots, which were irrigated under different regimes . For seeds buried at a depth of 50 mm, seed viability was examined after irrigation of the pots . * Key Results Seeds of three species germinated at most temperatures recorded between spring and autumn in their native habitats . No seed dormancy was found in any species . For all three species, seedling emergence was most favoured when seeds were buried at a depth of 10 mm . When seeds sown on the sand surface were irrigated, seed germination was considerably suppressed due to water deficiency, but many seeds remained viable . For A . squarrosum and B . dasyphylla, many seeds that were deeply buried and irrigated remained ungerminated but viable, while for A . adscensionis deeply buried seeds germinated, but the seedlings did not emerge due to unfavourable seedling growth in deep sand . * Conclusions Precipitation is the most crucial factor in determining the seasonal emergence of seedlings of the three tested species in the field . The vertical distribution of seeds in sand determines the proportion of seeds that germinate after precipitation and acts to maintain seed banks over multiple years. Ann Bot (Lond) . 2005 Jan 10; {Epub ahead of print} Effects of Three Nickel Salts on Germinating Seeds of Grevillea exul var . rubiginosa, an Endemic Serpentine Proteaceae; Leon V et al.; * Background and Aims Serpentine soils are usually quite infertile, arid and toxic, mainly because they contain high levels of heavy metals such as Ni . The aim of the present work was to assess the effects of Ni on the germinating seeds of Grevillea exul var . rubiginosa, an endemic serpentine Proteaceae of New Caledonia . In addition, the distribution of macronutrients and the Ni levels in germinating seeds were examined . * Methods Seeds were sown in glass Petri dishes and exposed to increasing concentrations of Ni (5 to 500 mg Ni L(-1)) using Ni chloride, Ni sulphate and Ni acetate . The germination percentage and root length were measured after 40 d . Longitudinal frozen sections of germinating seeds growing in the presence of Ni (500 mg L(-1) for all three salts) were used for X-ray microanalysis and X-ray elemental mapping using scanning electron microscopy (SEM) . * Key Results Ni chloride resulted in the greatest reductions in germination and root growth, particularly at 500 mg L(-1), followed by Ni sulphate and Ni acetate . SEM images revealed Ca crystalline structures in the seed coat for all the samples . S/Ca and Mg/P/K/Mn were found to be distributed differently in Ni-treated samples, whereas they all followed the same pattern in the controls . For all three salts, the Ni added to the medium had accumulated in the seed coat, whereas the endosperm seemed to be devoid of Ni . * Conclusions It is assumed that the seed coat is able to reduce the amount of Ni entering the seed, and that a high level of Ni induced the mobilization of macronutrients. In Vitro Cell Dev Biol Anim, 2004 Jul, 40(7), 196 - 210 MACROPHAGE POPULATION DYNAMICS WITHIN FETAL MOUSE FIBROBLAST CULTURES DERIVED FROM C57BL/6, CD-1, CF-1 MICE AND INTERLEUKIN-6 AND GRANULOCYTE COLONY STIMULATING FACTOR KNOCKOUT MICE; Talbot NC et al.; In vitro models of macrophage growth, differentiation, and function are needed to facilitate the study of their biology as important immune facilitator cells and as frequent targets of bacterial and viral infection . A simple method for the selective expansion and continuous culture of mouse macrophages from primary explant cultures of mouse embryonic tissue is described . Culture in Dulbecco modified Eagle medium (DMEM) low-glucose (1 g/L) formulation (DMEM/L) inhibited fibroblast growth . In contrast, macrophages continued to proliferate in the presence of DMEM/L when in contact with the fibroblasts . Alternating growth in high-glucose DMEM with DMEM/L produced a 1.16- to 2.1-fold increase (depending on mouse strain) in the percentage of macrophages within the cell culture in comparison with culturing in DMEM with high glucose exclusively . Macrophage yields of over 1 million cells/T12.5 flask were achieved by passages 3-4, and, thereafter, declined over the next 5-10 passages . The peak percentage of macrophages within a culture varied depending on the strain of mouse (C57BL/6, CD-1, and CF-1 and two knockout C57BL/6 strains deficient in either interleukin-6 {IL-6} or granulocyte colony stimulating factor {GCSF}) . The GCSF (-/-)-derived cultures had the lowest peak macrophage content (30%) and CD-1 the highest content (64.9%) . The IL-6 (-/-) and CD-1 cultures appeared to spontaneously transform to create cell lines (IL6MAC and CD1MAC, respectively) that were composed of 50-75% macrophages . The macrophages were phagocytic and were positive for CD14, acetylated low-density lipoprotein receptors, and F4-80 antigen . Light and electron microscopy showed that the cultured macrophages had in vivo-like morphological features, and they could be plated to high purity by differential attachment to petri dishes in serum-free medium. Biomed Mater Eng, 2005, 15(1-2), 101 - 12 In vitro cytotoxicity of amorphous carbon films; Rodil SE et al.; Amorphous carbon (a-C), carbon nitride (a-CN) and titanium films were deposited on stainless steel substrates (SS) using a dc magnetron sputtering system attached to a high vacuum chamber . Films were deposited using a base pressure of 1.3x10(-4) Pa . For the carbon films a pure graphite target was eroded in an Argon plasma . For the case of the a-CN films, the Ar flux was substituted by 100% N(2) gas . Titanium films were deposited in a different chamber, using a pure Ti target and an argon plasma . In vitro studies were carried out on the coated samples using human osteoblasts cells . Cytotoxicity of carbon films was assessed by cellular adhesion and proliferation, as determined by direct cellular counting using a spectroscopic technique and a well-defined standard curve . Osteoblasts cells were also grown on uncoated steel and prepared Petri dishes for comparison . The percentage of osteoblasts adhesion measured at 24 hrs attained maximum values for the a-C films . Similarly, cellular proliferation evaluated at three, five and seven days showed an outstanding increase of osteoblasts cells for the a-C and Ti coatings in contrast to the uncoated steel . The cell functionality was evaluated by the MTT test after incubation periods of 3, 5 and 7 days . The absorbance values obtained for a-C, a-CN and Ti surfaces resulted significantly higher with respect to the positive control, indicating that the surface did not induce any toxic effect . Preliminary bio-mineralization was evaluated by measuring the elemental composition of the mineral grown on the substrates after periods up to 14 days. Biofouling, 2004 Aug-Oct, 20(4-5), 211 - 7 Settlement Behaviour of Marine Invertebrate Larvae Measured by EthoVision 3.0; Marechal JP et al.; Submerged marine surfaces are rapidly colonized by fouling organisms . Current research is aimed at finding new, non-toxic, or at least environmentally benign, solutions to this problem . Barnacles are a major target organism for such control as they constitute a key component of the hard fouling community . A range of standard settlement assays is available for screening test compounds against barnacle cypris larvae, but they generally provide little information on mechanism(s) of action . Towards this end, a quick and reliable video-tracking protocol has been developed to study the behaviour of the cypris larvae of the barnacle, Balanus amphitrite, at settlement . EthoVision 3.0 was used to track individual cyprids in 30-mm Petri dishes . Experiments were run to determine the optimal conditions vis-a-vis acclimation time, tracking duration, number of replicates, temperature and lighting . A protocol was arrived at involving a two Petri dish system with backlighting, and tracking over a 5-min period after first acclimating the cyprids to test conditions for 2 min . A minimum of twenty replicates was required to account for individual variability in cyprid behaviour from the same batch of larvae . This methodology should be widely applicable to both fundamental and applied studies of larval settlement and with further refinements, to that of smaller fouling organisms such as microalgae and bacteria. Mycol Res, 2004 Nov, 108(Pt 11), 1241 - 56 Simulating colonial growth of fungi with the Neighbour-Sensing model of hyphal growth; Meskauskas A et al.; The Neighbour-Sensing model brings together the basic essentials of hyphal growth kinetics into a vector-based mathematical model in which the growth vector of each virtual hyphal tip is calculated by reference to the surrounding virtual mycelium . The model predicts the growth pattern of many hyphae into three spatial dimensions and has been used to simulate complex fungal fruit body shapes . In this paper we show how the Neighbour-Sensing model can simulate growth in semi-solid substrata like agar or soil, enabling realistic simulation of mycelial colonies of filamentous fungi grown in 'Petri-dish style' experimental conditions . Newly implemented capabilities in the model include: a measurement and logging system within the program that maintains basic statistics about the mycelium it is simulating, this facilitates kinetic experimentation; inclusion of 'substrates' in the data space causing positive or negative tropisms for the growing mycelium; a horizontal plane tropism that provides a way of simulating colonies growing in or on a substratum like agar or soil by imposing a horizontal constraint on the data space the cyberhyphal tips can explore; three categories of hypha--standard hyphae are those that start the simulation, leading hyphae can emerge from the colony peripheral growth zone to take on a leading role, and secondary hyphae are branches that can arise late, far behind the peripheral growth zone, when mature hyphal segments resume branching to in-fill the older parts of the colony . We show how the model can be used to investigate hyphal growth kinetics in silico in experimental scenarios that would be difficult or impracticable in vivo . We also show that the Neighbour-Sensing model can generate sufficiently realistic cord-like structures to encourage the belief that this model is now sufficiently advanced for parameters to be defined that simulate specific in silico cyberfungi . The potential utility of these cyberspecies is that they provide a means to model the morphogenetic effects of a variety of factors, from environmental and nutritional features to mutations, in experimentally realistic situations, offering a valuable addition to the experimental toolkit of all those interested in fungal growth and morphology. J Chem Ecol, 2004 Sep, 30(9), 1781 - 95 Attack rate and success of the parasitoid Diaeretiella rapae on specialist and generalist feeding aphids; Blande JD et al.; Lipaphis erysimi (Kaltenbach) is a specialist crucifer feeding aphid and Myzus persicae (Sulzer) is a generalist feeding aphid . The foraging behavior of Diaeretiella rapae (McIntosh), a parasitoid with the ability to parasitize both of these species, was assessed using a series of attack rate and success bioassays, with turnip, Brassica rapa var rapifera, as the host plant . The attack rate of D . rapae was significantly greater on L . erysimi than on M . persicae when aphids were feeding on turnip leaf discs in Petri dishes, irrespective of the aphid species upon which the parasitoids were originally reared . Attack rate bioassays with leaf discs absent, using both satiated and starved aphids, revealed that background chemistry and internal aphid chemistry may have small effects on attack rate . Excision of D . rapae pupae from mummy cases and subsequent use of the fully developed adults in attack rate bioassays showed that cues received by D . rapae at the time of adult emergence provide cues that prime D . rapae to attack L . erysimi at a greater rate than M . persicae . However, the relative success of D . rapae on these two aphid species, in terms of the percentage of attacks resulting in a successful adult parasitoid, was not significantly different. Cell Mol Life Sci, 2004 Dec, 61(24), 3093 - 9 Acute effects of adrenergic agents on post-defibrillation arrest time in a cultured heart model; Krauthamer V et al.; Possible drug interactions with electrical defibrillation were examined . We tested the hypothesis that adrenergic agents (epinephrine, norepinephrine, isoproterenol) and a calcium channel blocker (verapamil), when applied acutely, alter the duration of arrest following a defibrillator shock . A secondary hypothesis (based on observations) was that the drugs alter the occurrence of changes to normal rhythms following the shock . Dissociated heart cells from 10-day chicken embryos were cultured to form spherical aggregates and plated in petri dishes . In the experiments, the spheres were paced at 0.75 V/cm above contraction threshold, and a biphasic defibrillator shock was applied for 1 ms at 46 V/cm . The arrest time and occurrence of rhythm changes were recorded . The adrenergic agents shortened the duration of arrest following a defibrillator shock, while the calcium channel blocker lengthened the arrest time . Comparisons with the control proportion of double beats showed no significant change with the adrenergic agents and a decrease with verapamil. Chem Biol Interact, 2004 Dec 7, 150(3), 211 - 9 The activities of several detoxication enzymes are differentially induced by juices of garden cress, water cress and mustard in human HepG2 cells; Lhoste EF et al.; It has been previously demonstrated in a human-derived hepatoma cell line (HepG2) that juices from cruciferous vegetables protect against the genotoxicity caused by dietary carcinogens . HepG2 cells possess different enzymes involved in the biotransformation of xenobiotics . Therefore, we investigated the effect of cruciferous juices on the activities of CYP 1A and several phase II enzymes in this cell model . For each experiment, 1 x 10(6) cells were seeded on Petri dishes . After 2 days, the juices (0.5-8 microl/ml of culture medium) were added for 48 h prior to cell harvesting . The addition of juice from water cress (Nasturtium officinalis R . Br) significantly increased the activities of ethoxyresorufin-O-deethylase at high doses only and NAD(P)H-quinone reductase in a dose-dependent manner (1.8- and 5-fold, respectively) . The addition of juice from garden cress (Lepidum sativum L.) significantly increased the activities of NAD(P)H-quinone reductase and UDP-glucuronosyl-transferase with a maximal effect around the dose of 2 microl/ml juice (1.4- and 1.2-fold, respectively) while the other enzymes were not altered . Mustard (Sinapis alba L.) juice increased the activities of NAD(P)H-quinone reductase (2.6-fold at the dose of 8 microl/ml), and N-acetyl-transferase (1.4-fold at the dose of 8 microl/ml) in a dose-dependent manner while a maximal induction of UDP-glucuronosyl-transferase was obtained with a dose of 2 microl/ml (1.8-fold) . These observations show that the three juices have different induction profiles: only water cress acted as a bifunctional inducer by enhancing both phase I and phase II enzymes . As a consequence, each juice may preferentially inhibit the genotoxicity of specific compounds. Invest Ophthalmol Vis Sci, 2004 Dec, 45(12), 4320 - 6 Induction of epithelial progenitors in vitro from mouse embryonic stem cells and application for reconstruction of damaged cornea in mice; Homma R et al.; PURPOSE: Severe ocular surface diseases and injuries cause loss of the corneal limbal epithelium, leading to re-epithelialization by bulbar conjunctival cells, resulting in vascularization of the cornea, conjunctival scarring, and loss of visual acuity . In this study, the optimal culture condition for induction of differentiation of epithelial progenitor cells from embryonic stem (ES) cells was determined for use in transplantation to damaged cornea in mice . METHODS: Mouse ES cells were cultured on Petri dishes coated with several extracellular matrix proteins, and the markers for epithelial cells were analyzed with RT-PCR and Western blot analysis . The optimal condition for induction of epithelial progenitor cells was determined, and the progenitors were transplanted onto mouse eyes with corneal epithelia that had been damaged by exposure to n-heptanol . RESULTS: Epithelial progenitors were successfully induced by culturing mouse ES cells on type IV collagen for 8 days . These progenitors expressed keratin (K)12, which is specific to corneal epithelial cells, and cell surface CD44 and E-cadherin, both of which are essential in corneal epithelial wound healing . Complete re-epithelialization of the corneal surface occurred within 24 hours after transplantation . The resultant corneal epithelial cells expressed markers of the grafted cells, and no teratomata were observed during the follow-up period . CONCLUSIONS: Epithelial progenitors were successfully induced in vitro from ES cells and were applicable as grafts for treating corneal epithelial injury . ES cells may become an unlimited donor source of corneal epithelial cells for corneal transplantation and may restore useful vision in patients with a deficiency of limbal epithelial cells . This is an important first trial toward assessing the use of ES cells to reconstruct corneal epithelial cells. Biomacromolecules, 2004 Nov-Dec, 5(6), 2094 - 104 Effects on interfacial properties and cell adhesion of surface modification by pectic hairy regions; Morra M et al.; Polystyrene Petri dishes, aminated by a plasma deposition process, were surface modified by the covalent linking of two different enzymatically modified hairy regions (HRs) from pectin containing, for example, rhamnogalacturonan-I and xylogalacturonan structural elements . The two polysaccharide preparations share the same structural elements of apple pectin, but the relative amounts and lengths of the neutral side chains present differ . Surface analysis by X-ray photoelectron spectroscopy, contact angle measurement, and atomic force microscope (AFM) force-separation curves was used to characterize the effects on surface chemistry and interfacial forces of the surface modification process . Cell adhesion experiments using continuous L-929 fibroblasts and primary aortic smooth muscle cells were performed to evaluate the effect of the polysaccharide nature on cell adhesion . Results show that immobilization of the HR affects the interfacial field of forces and the cell behavior: "equilibrium" contact angles, obtained by a recently introduced vibrational approach, decrease after HR immobilization reaching a value close to 20 degrees . AFM force-separation curves show a more extended (or softer) interface in the case of the HR bearing longer side chains . Accordingly, depending on the HR preparation, cells shifted from spread morphology and adhesion behavior quantitatively comparable to that observed on conventional tissue culture polystyrene to rounded morphology and significantly lower adhesion . These data show that engineering of plant pectins can be a valuable tool to prepare novel and finely tuned polysaccharides having different chemico-physical and biological properties, to be used in the surface modification of medical devices and materials. Phys Rev E Stat Nonlin Soft Matter Phys . 2004 Sep;70(3 Pt 1):031912 . Epub 2004 Sep 28. Periodically varying externally imposed environmental effects on population dynamics; Ballard M et al.; Effects of externally imposed periodic changes in the environment on population dynamics are studied with the help of a simple model . The environmental changes are represented by the temporal and spatial dependence of the competition terms in a standard equation of evolution . Possible applications of the analysis are on the one hand to bacteria in Petri dishes and on the other to rodents in the context of the spread of the Hantavirus epidemic . The analysis shows that spatiotemporal structures emerge, with interesting features which depend on the interplay of separately controllable aspects of the externally imposed environmental changes. Radiat Prot Dosimetry, 2004, 112(3), 419 - 28 Epub 2004 Oct 19. 1800 MHz in vitro exposure device for experimental studies on the effects of mobile communication systems; Ardoino L et al.; A wire patch cell (WPC) operating at the uplink frequency band of GSM 1800 MHz has been designed for in vitro experiments with the aim of investigating the possible biological effects of electromagnetic radiation associated with cellular phones . The 1800 MHz WPC design is a direct descendant of the original 900 MHz WPC introduced by Laval et al . This system provides a homogeneous specific absorption rate distribution, using four 3.5 cm petri dishes simultaneously . Numerical dosimetry has been performed using a commercial code (CST Microwave Studio), in order to evaluate accurately the efficiency of the structure (in terms of W kg(-1) per 1 W input power) and the distribution in the chosen biological target . The numerical results have been confirmed by experimental measurements performed by measuring thermal increase due to a high power impulse . The efficiency of the structure is 1.25 +/- 25% W kg(-1) per 1 W input power higher than the efficiency of the 900 MHz WPC . A few adjustments have been made in order to use the WPC in a standard incubator and to avoid thermal increases related to the radio frequency exposure . This exposure system has been adopted for the experiments scheduled in the RAMP and GUARD projects (VFPE). Parasitol Res, 2004 Oct, 94(3), 201 - 6 Epub 2004 Sep 01. Steinernema glaseri Santa Rosa strain (Rhabditida: Steinernematidae) and Heterorhabditis bacteriophora CCA Strain (Rhabditida: Heterorhabditidae) as biological control agents of Boophilus microplus (Acari: Ixodidae); de Oliveira Vasconcelos V et al.; The present study was carried out to evaluate the action of Steinernema glaseri Santa Rosa strain and Heterorhabditis bacteriophora CCA strain as biological control agents of Boophilus microplus . Engorged females ticks were distributed on Petri dishes containing different concentrations of infective juvenile (IJ) nematodes (0, 375, 500, 750, 1,500, 2,500, 5,000 and 25,000) . The data showed a reduction of approximately 90% in the eggs laid at a concentration of 5,000 S . glaseri IJs and approximately 80% at a concentration of 1,500 H . bacteriophora IJs . The female mortality increased linearly with the increase in S . glaseri concentrations . However, in the tests with H . bacteriophora this linearity was not observed . The effectiveness of the treatment with both species of entomopathogenic nematodes was compatible with other control methods . The results show the potential of S . glaseri and H . bacteriophora as biological control agents for the control of B . microplus under laboratory conditions. Microsc Res Tech, 2004 Jun 15, 64(3), 228 - 42 Multidimensional long-term time-lapse microscopy of in vitro peripheral nerve regeneration; Ozturk G et al.; In order to test the effectiveness of a new advanced time-lapse microscopy imaging and image processing and analysis system, and to do quantitative and qualitative temporal analyses of in vitro peripheral nerve regeneration, long-term time-lapse imaging of cultures of mouse dorsal root ganglia (DRGs) was performed . DRGs were placed in a Petri dish, covered with collagen gel, their attached peripheral nerves were cut in the middle, creating a gap, and the dish was filled with culture medium . Six preparations were kept on the time-lapse imaging system, which provides a suitable incubation environment and enables to capture images from multiple coordinates at x,y,z axes at desired time intervals for 13 days . In general, the time-lapse imaging system proved quite stable and efficient, although some improvements are certainly required . Two main components of peripheral nerve regeneration, outgrowth of axons and activities of resident cells, were examined . Axons started to grow during the first hour of incubation with a 16.5 microm/h rate and showed the slowest rates (0.7 microm/h) on days 8 and 9, after which they resumed higher speeds again . The first cell came out of the proximal end of the cut nerve on the second day and it was a Schwann cell (SC), which was the prominent cell type in the preparations throughout the experiment . SCs were higher in number (83.15% of all cells) but slower in migration (3.4 vs . 7.3 microm/h, P < 0.001) than other cells . Other observed characteristics of axonal outgrowth and cellular activity and interactions between axons and the cells are discussed . J Econ Entomol, 2004 Aug, 97(4), 1239 - 44 Effects of insect growth regulators on citrus mealybug parasitoid Leptomastix dactylopii (Hymenoptera: Encyrtidae); Rothwangl KB et al.; In this study, we measured the effects of three commonly used insecticides classified as insect growth regulators, on the encyrtid parasitoid Leptomastix dactylopii (Howard) when parasitizing citrus mealybug, Planococcus citri (Risso) . Kinoprene, pyriproxyfen, and azadirachtin were evaluated in both petri dish and a cage experiment at label-recommended rates to measure their effects on the mortality, parasitization rate, and sex ratio of L . dactylopii . Insecticides were applied to petri dishes and plants either immediately before, 24 h before, or 48 h before release of the parasitoid . Kinoprene applied 24 h before parasitoid release caused 100% mortality of L . dactylopii in petri dishes within 48 h . Mortality rates for L . dactylopii exposed to azadirachtin and pyriproxyfen did not exceed 5% regardless of release time . There were no release time x insecticide interactions on L . dactylopii parasitization rate . The insecticide alone, however, did significantly affect parasitization rates of L . dactylopii on P . citri; the kinoprene treatment significantly reduced L . dactylopii parasitization rates compared with azadirachtin and pyriproxyfen . In a cage experiment with coleus, Solenostemon scutellaroides (L.) Codd, applications of both pyriproxyfen and kinoprene resulted in fewer P . citri parasitized by L . dactylopii than azadirachtin or the control . The sex ratio was equivalent in the petri dish experiment, whereas in the cage experiment the sex ratio was biased toward males, particularly for the kinoprene treatment . Based on the results from this study, we suggest that kinoprene is not compatible with releases of L . dactylopii to control citrus mealybugs. Cancer Res, 2004 Sep 15, 64(18), 6438 - 43 Functional characterization of the candidate tumor suppressor gene NPRL2/G21 located in 3p21.3C; Li J et al.; Initial analysis identified the NPRL2/G21 gene located in 3p21.3C, the lung cancer region, as a strong candidate tumor suppressor gene . Here we provide additional evidence of the tumor suppressor function of NPRL2/G21 . The gene has highly conserved homologs/orthologs ranging from yeast to humans . The yeast ortholog, NPR2, shows three highly conserved regions with 32 to 36% identity over the whole length . By sequence analysis, the main product of NPRL2/G21 encodes a soluble protein that has a bipartite nuclear localization signal, a protein-binding domain, similarity to the MutS core domain, and a newly identified nitrogen permease regulator 2 domain with unknown function . The gene is highly expressed in many tissues . We report inactivating mutations in a variety of tumors and cancer cell lines, growth suppression of tumor cells with tet-controlled NPRL2/G21 transgenes on plastic Petri dishes, and suppression of tumor formation in SCID mice . Screening of 7 renal, 5 lung, and 7 cervical carcinoma cell lines showed homozygous deletions in the 3' end of NPRL2 in 2 renal, 3 lung, and 1 cervical (HeLa) cell line . Deletions in the 3' part of NPRL2 could result in improper splicing, leading to the loss of the 1.8 kb functional NPRL2 mRNA . We speculate that the NPRL2/G21 nuclear protein may be involved in mismatch repair, cell cycle checkpoint signaling, and activation of apoptotic pathway(s) . The yeast NPR2 was shown to be a target of cisplatin, suggesting that the human NPRL2/G21 may play a similar role . At least two homozygous deletions of NPRL2/G21 were detected in 6 tumor biopsies from various locations and with microsatellite instability . This study, together with previously obtained results, indicates that NPRL2 is a multiple tumor suppressor gene. J Mater Sci Mater Med, 1997, 8(10), 587 - 90 Phenotypic characterization of keratinocytes migrated from polymer support - in vitro study; SMETANA Jr K et al.; The keratinocytes are able to migrate from the poly (2- hydroxyethylmethacrylate) disc if it is transfered to the new Petri-dish colonized with irradiated 3T3 mouse fibroblasts, and form a ring-shaped colony around the disc . The phenotypic characterization of human keratinocytes migrated from these discs was studied using a group of monoclonal antibodies . The keratinocytes in the external periphery of the colony of cells which migrated from the disc express the proliferating cell nuclear antigen (PCNA), alpha2, alpha3 chains and alpha5beta1 integrin receptor . A protein of the desmosome complex, desmoplakin-1, was also expressed . Involucrin and cytokeratin-10 were expressed after prolonged cultivation . These results suggest that the migrated keratinocytes are able to proliferate, recognize extracellular matrix molecules important in the process of the re-epitelization of the wound, and terminally differentiate in vitro . They are encouraging for further experiments with respect to the development of a support for keratinocyte cultivation and for grafting in clinical practice. J Mater Sci Mater Med, 2001 Dec, 12(10/12), 997 - 1000 Osteoblast viability and detachment following exposure to ultrasound in vitro; Sura H et al.; Ultrasound has been used in dentistry for over 40 years and has recently been proposed for cutting bone . The purpose of this study was to establish the effects of ultrasonic instruments on osteoblasts . A 25 kHz magnetostrictive ultrasound generator and a TFI-1 tip (Dentsply, UK) were used as the ultrasound generating instruments . Primary osteoblast cultures were established from the parietal bones of two-day-old Albino Wistar rats grown on tissue culture (TC) petri dishes (Corning, UK) in alphaMEM (Sigma, UK) . Once confluent, the osteoblasts were harvested using 0.05% trypsin in 0.02% EDTA then 1.7\times 10^{5} cells in 2.5 ml of alphaMEM were either re-seeded immediately onto TC dishes and allowed to adhere for 24 h or kept in suspension before application of ultrasound with different tip displacements prior to re-seeding the cells . Osteoblast viability was assessed using 0.4% Trypan Blue following the initial dose of ultrasound then periodically over a 20 h period for both adherent and suspension osteoblasts . This study demonstrated that ultrasound caused osteoblast detachment and loss of viability in vitro, both when adherent to a substrate or in suspension . Loss of osteoblast viability was related to the maximum displacement of the ultrasonic tip and continued throughout the 20 h period observed for osteoblasts adherent to TC dishes. J Mater Sci Mater Med, 2000 Dec, 11(12), 773 - 8 Cellular responses to chitosan in vitro: The importance of deacetylation; Prasitsilp M et al.; Chitin and chitosan (a deacetylated derivative of chitin) have been proposed for biomedical applications because of their biocompatibility and abundance in nature . We have investigated the effect of the percentage of deacetylation (%DD) of chitosan on biocompatibility from two sources, shrimp and cuttle fish, with two cell lines, L929 and BHK21(C13) . The difference in %DD for each source was approximately 10% in the range of 76-90% . Biocompatibility was investigated for: (1) cell adherence and growth on the chitosan samples as substrate; (2) the effect of extract media on 2d and 7d growth; and (3) the presence of an inhibition zone . The results were similar for both cell lines . The chitosan samples were air-dried on to tissue culture-grade petri dishes to provide a substrate for the adherent-cell cultures . The higher %DD substrates from each source supported attachment of the cells, while the lower %DD did not . Cells cultured in medium conditioned by each substrate (i.e . extract medium) displayed an initial difference in growth which was abrogated in cultures incubated for 7 days . No inhibition zone was apparent . However, after 7 days, some cells were noted migrating on to the low %DD substrate disks . The morphology of these cells was changed with the presence of pseudopodia being apparent . Thus, especially with regard to attachment the %DD has a very important effect on the biocompatibility of the chitosan and should be monitored carefully . J Mater Sci Mater Med, 2004 Jun, 15(6), 693 - 7 Osteoblast response and calcium deposition on phospholipid modified surfaces; Satsangi N et al.; In this study, the effect of calcium phosphate complexed phospholipid (Ca-PL-PO4) coatings on solid surfaces on the in vitro calcium (Ca) deposition and on the osteoblast responses was evaluated . Commercially available phospholipids were converted to their Ca-PL-PO4, and were coated on glass Petri dishes . The coated dishes were immersed in the simulated body fluid for up to 14 days under sterilized conditions at 37 degrees C, and the amount of calcium (Ca) deposited was quantified . Similarly, by measuring the alkaline phosphatase specific activity, the differentiation of osteoblast precursor cells were evaluated after seeding the cells on Ca-PL-PO4 coated cell culture plastics . It was observed that all Ca-PL-PO4 enhanced Ca deposition on coated surfaces . The, polar head group of phospholipids in coated surfaces was observed to have an influence on the Ca deposition as well as the osteoblast differentiation . Among the phospholipids evaluated, phosphatidylserine (Ca-PS-PO4) exhibited the strongest calcium deposition and more enhanced alkaline phosphatase specific activity . It was therefore concluded from this study that Ca-PS-PO4 surface modification may be an alternative method for enhancing bone-implant interactions. Allergol Immunopathol (Madr), 2004 Jul-Aug, 32(4), 197 - 203 Indoor airborne fungal spores and home characteristics in asthmatic children from Edirne region of Turkey; Yazicioglu M et al.; BACKGROUND: The contribution of indoor fungal exposure to childhood asthma is not completely clear . OBJECTIVE: To investigate airborne fungal flora within the homes of asthmatic and control children, and to assess the influence of housing characteristics regarding indoor fungi . METHODS: Forty-seven atopic asthmatic and 23 nonatopic control children were studied . Allergen sensitivity was determined by skin prick tests . A thorough assessment, using a questionnaire and inspection surveys, was carried out . Home visits were made between October 2000 and February 2001 . Samples of airborne fungal spores were collected from four rooms by the "open Petri dish" method . Indoor temperature and humidity were measured . RESULTS: The total indoor fungal colony counts from the living rooms and bedrooms were significantly higher in the asthma group than in controls (p = .012 and p = .003, respectively) . The most commonly isolated genus was Cladosporium . Twelve of the asthmatic patients (25.53 %) were found to be sensitive to fungal allergens . The factors found to be associated with indoor fungal growth in logistic regression were visible fungal patches in the bathrooms {(odds ratio (OR) = 5.75; 95 % CI 1.19 to 27.70)}, and the age of the house {OR = 4.24; 95 % CI 1.34 to 13.45} . Total fungal colony numbers did not correlate with indoor temperature or humidity . CONCLUSION: Fungal colony numbers were higher in the homes of asthmatic children than in those of controls . Therefore, indoor fungal exposure may contribute to childhood asthma . Bathrooms were the main source of fungal propagules . Old houses were more prone to fungal growth. J Neurophysiol, 2005 Jan, 93(1), 580 - 93 Epub 2004 Aug 18. Quantitative characterization and classification of leech behavior; Mazzoni A et al.; This paper describes an automatic system for the analysis and classification of leech behavior . Three colored beads were attached to the dorsal side of a free moving or pinned leech, and color CCD camera images were taken of the animal . The leech was restrained to moving in a small tank or petri dish, where the water level can be varied . An automatic system based on color processing tracked the colored beads over time, allowing real-time monitoring of the leech motion for several hours . At the end of each experimental session, six time series (2 for each bead) describing the leech body motion were obtained . A statistical analysis based on the speed and frequency content of bead motion indicated the existence of several stereotypical patterns of motion, corresponding to different leech behaviors . The identified patterns corresponded to swimming, pseudo-swimming, crawling, exploratory behavior, stationary states, abrupt movements, and combinations of these behaviors . The automatic characterization of leech behavior demonstrated here represents an important step toward understanding leech behavior and its properties . This method can be used to characterize the behavior of other invertebrates and also for some small vertebrates. Am J Physiol Cell Physiol, 2004 Dec, 287(6), C1527 - 36 Epub 2004 Aug 18. "Culture shock" from the bone cell's perspective: emulating physiological conditions for mechanobiological investigations; Sorkin AM et al.; Bone physiology can be examined on multiple length scales . Results of cell-level studies, typically carried out in vitro, are often extrapolated to attempt to understand tissue and organ physiology . Results of organ- or organism-level studies are often analyzed to deduce the state(s) of the cells within the larger system(s) . Although phenomena on all of these scales-cell, tissue, organ, system, organism-are interlinked and contribute to the overall health and function of bone tissue, it is difficult to relate research among these scales . For example, groups of cells in an exogenous, in vitro environment that is well defined by the researcher would not be expected to function similarly to those in a dynamic, endogenous environment, dictated by systemic as well as organismal physiology . This review of the literature on bone cell culture describes potential causes and components of cell "culture shock," i.e., behavioral variations associated with the transition from in vivo to in vitro environment, focusing on investigations of mechanotransduction and experimental approaches to mimic aspects of bone tissue on a macroscopic scale . The state of the art is reviewed, and new paradigms are suggested to begin bridging the gap between two-dimensional cell cultures in petri dishes and the three-dimensional environment of living bone tissue. J Chem Ecol, 2004 Jun, 30(6), 1257 - 69 Chemical and physical signals mediating conspecific and heterospecific aggregation behavior of first instar stink bugs; Fucarino A et al.; We investigated cues that mediate the aggregation behavior of immature pentatomid bugs by using nymphs of six different pentatomid bug species (Nezara viridula, Acrosternum hilare, Chlorochroa ligata, Chlorochroa sayi, Thyanta pallidovirens, and Euschistus conspersus) . When first instars of any two species were put together in a Petri dish, they readily formed heterospecific aggregations similar to their natural conspecific aggregations . The chemical profiles of first and second instar nymphs of each species were determined by solvent extraction with pentane, followed by GC-MS analysis . Immature bugs of the different species had some compounds in common, and some that were more species specific . Within a species, there were distinct differences in the profiles of compounds extracted from first and second instars . Bugs did not aggregate around untreated polysulfone beads (1 mm diam) that were glued together in groups approximating bug egg masses, suggesting that tactile cues alone were insufficient to induce aggregation . Furthermore, when tested over a range of doses, groups of polysulfone beads treated with crude whole-body extracts of bugs did not induce or maintain aggregations . However, first instar N . viridula nymphs did respond to beads treated with two of the three major components of bug extracts . 4-Oxo-(E)-2-decenal induced significant aggregations at two doses, whereas tridecane, the major component in extracts from all six species, did not, and (E)-2-decenal was repellent . The repellence of (E)-2-decenal may explain why we and previous researchers were unable to induce aggregations of first instar N . viridula using whole-body extracts. J Chem Ecol, 2004 Jun, 30(6), 1103 - 16 The role of semiochemicals in the avoidance of the seven-spot ladybird, Coccinella septempunctata, by the aphid parasitoid, Aphidius ervi; Nakashima Y et al.; The role of semiochemicals in mediating intraguild interactions between the seven-spot ladybird, Coccinella septempunctata, and the aphid parasitoid, Aphidius ervi, was investigated . Female parasitoids avoided leaves visited by C . septempunctata adults and larvae during the previous 24 hr . Ethanol extracts of C . septempuntata adults and larvae also induced avoidance responses by A . ervi . Two of the hydrocarbons identified by gas chromatography (GC) and coupled GC-mass spectrometry (GC-MS), n-tricosane (C23H48), and n-pentacosane (C25H52), when tested individually at levels found in the adult extract, induced avoidance by A . ervi . Further investigation of the larvae extract, and footprint chemicals deposited by adults in glass Petri dishes, confirmed the presence of the hydrocarbons . Parasitism rates of the pea aphid, Acyrthosiphon pisum, on broad bean plants, Vicia faba, which had been sprayed with a mixture of the chemicals, were lower than those on control plants . The effect, however, was no longer evident if parasitoid foraging was delayed by 24 hr after the plants were treated . The ecological significance of intraguild avoidance behavior and implications for possible use of the semiochemicals involved in future biological control programs are discussed. Environ Sci Technol, 2004 Jul 15, 38(14), 3901 - 7 Rapid release of mercury from intertidal sediments exposed to solar radiation: a field experiment; Canario J et al.; There is increasing evidence of the primary importance of photochemical reactions and transfer of gaseous mercury to the atmosphere . Although mercury in aquatic sediments is efficiently retained, resuspension and bioturbation in intertidal sediments may expose temporarily anoxic sediments to solar radiation . Field experiments were performed to investigate these processes . Anoxic sediments from two areas in the Tagus estuary with different degrees of Hg contamination (experiments I and II) were homogenized and distributed into two sets of 36 uncovered Petri dishes . The samples were placed on the intertidal sediments and exposed to direct solar radiation and kept under dark (control) for 6-8 h . The decrease rates of acid volatile sulfides (abrupt in the first 3 h) and of pyrite (linear) were the same in sediments under solar radiation and dark . The total Hg concentrations were relatively constant in sediments kept in dark, but decreased from 17.6 to 7.65 and 3.45 to 1.35 nmol g(-1) in experiments I and II, respectively . In those exposed to solar radiation during the period of higher UV intensity . Similar evolutions were found in nonreactive Hg in pore waters (3.00-2.59 and 0.725-0.105 nM) . On the contrary, reactive Hg was higher in pore waters of the sediments exposed to solar radiation and increased with time, from 424 to 845 pM and 53 to 193 pM . These results indicate that most mercury released in pore waters was photochemically reduced in a short period of time and escaped rapidly to the atmosphere . Episodes of bottom resuspension and bioturbation in the intertidal sediments enhance the transfer of gaseous mercury to the atmosphere. Cytometry, 2004 Aug, 60A(2), 182 - 8 Automated counting of mammalian cell colonies by means of a flat bed scanner and image processing; Dahle J et al.; BACKGROUND: Clonogenic assays are used frequently to measure the cell killing and mutagenic effects of radiation and other agents . Clonogenic assays carried out manually are tedious and time-consuming and involve a significant element of subjectivity . However, several commercial automatic colony counters are available . Based on CCD video imaging and image analysis they are relatively expensive and can analyze only one petri dish at a time . METHOD: We have developed a cheaper and more efficient device, which employs a flat bed scanner to image 12 60-mm petri dishes at a time . Two major problems in automated colony counting are the clustering of colonies and edge effects . By using standard image analysis and implementing an inflection point algorithm, these problems were greatly diminished . The resulting system was compared with two manual colony counts, as well as with automated counts with the Oxford Optronix ColCount colony counter for cell lines V79 and HaCaT . RESULTS: Comparisons assuming the manual counts to be correct showed that our automatic counter was slightly more accurate than the commercial unit . CONCLUSIONS: As a whole, our automated colony counter performed significantly better than the commercial unit with regard to processing time, cost and accuracy. Rev Inst Med Trop Sao Paulo, 2004 May-Jun, 46(3), 133 - 7 Epub 2004 Jul 20. Airborne fungi isolated from Fortaleza city, State of Ceará, Brazil; Menezes EA et al.; Airbone fungi are considered important causes of allergic rhinitis and allergic asthma . The knowledge of these fungi in a city or region is important for the ecological diagnosis and specific treatment of allergic manifestations induced by inhalation of fungal allergens . The airborne fungi of Fortaleza, State of Ceara, Brazil, were studied during a one year period . Five hundred and twenty Petri dishes with Sabouraud dextrose agar medium were exposed at ten different locations in the city . The dishes exposed yielded one thousand and five hundred and twenty one colonies of twenty four genera . The most predominants were: Aspergillus (44.7%), Penicillium (13.3%), Curvularia (9.8%), Cladosporium (6.8%), Mycelia sterilia (6.0%), Fusarium (3.5%), Rhizopus (3.1%), Drechslera (2.6%), Alternaria (2.4%) and Absidia (2.2%) . The results shown that Aspergillus, Penicillium, Mycelia sterilia, Fusarium and Alternaria were found during all months in the year . Absidia was more frequent during the dry season . Anemophilous fungi and the high concentration of spores in the air are important because may result in an increased number of people with allergic respiratory disease. Phytochemistry, 2004 Jun, 65(12), 1817 - 28 A proteomic approach to studying plant response to crenate broomrape (Orobanche crenata) in pea (Pisum sativum); Angeles Castillejo M et al.; Crenate broomrape (Orobanche crenata) is a parasitic plant that threatens legume production in Mediterranean areas . Pea (Pisum sativum) is severely affected, and only moderate levels of genetic resistance have so far been identified . In the present work we selected the most resistant accession available (Ps 624) and compared it with a susceptible (Messire) cultivar . Experiments were performed by using pot and Petri dish bioassays, showing little differences in the percentage of broomrape seed germination induced by both genotypes, but a significant hamper in the number of successfully installed tubercles and their developmental stage in the Ps 624 compared to Messire . The protein profile of healthy and infected P . sativum root tissue were analysed by two-dimensional electrophoresis . Approximately 500 individual protein spots could be detected on silver stained gels . At least 22 different protein spots differentiated control, non-infected, Messire and Ps 624 accessions . Some of them were identified by MALDI-TOF mass spectrometry and database searching as cysteine proteinase, beta-1,3-glucanase, endochitinase, profucosidase, and ABA-responsive protein . Both qualitative and quantitative differences have been found among infected and non-infected root extracts . Thus, in the infected susceptible Messire genotype 34 spots were decreased, one increased and three newly detected, while in Ps 624, 15 spots were increased, three decreased and one newly detected . In response to the inoculation, proteins that correspond to enzymes of the carbohydrate metabolism (fructokinase, fructose-bisphosphate aldolase), nitrogen metabolism (ferredoxin-NADP reductase) and mitochondrial electronic chain transport (alternative oxidase 2) decreased in the susceptible check, while proteins that correspond to enzymes of the nitrogen assimilation pathway (glutamine synthetase) or typical pathogen defence, PR proteins, including beta-1,3-glucanase and peroxidases, increased in Ps 624 . Results are discussed in terms of changes in the carbohydrate and nitrogen metabolism an induction of defence proteins in response to broomrape parasitism. Indian J Exp Biol, 2003 Jun, 41(6), 563 - 9 Tissue engineering: in vitro embryonal nidation in a murine endometrial construct; Stevenson AF; The epithelial and mesothelial cellular components of organs can be obtained as dissociated cells using adequate procedures of enzymatic digestion followed by pycnotic separation on density gradients . Using a specially developed procedure for tissue dissociation, the epithelial and connective tissue components of endometria from pseudopregnant mice were grown in culture using a combination of three dimensional culture of connective tissue components in collagen gel, with the superimposition of epithelial components in liquid medium on the surface of the gells . After a few days of growth, when the cultures became dense, murine blastocysts obtained on postcoital day 4.5 by fallopian flushing of hormonally primed and mated mice, were transferred onto the imitated endometria . The blastocysts hatched and grew on the endometrial epithelium as spherical coherent conglomerates of cells quite different from hatched blastocysts grown on the surface of a petri dish, in which the presumtive trophoblasts spred around the central mass . Light and electronmicroscopy of resin embeded sections (2 days after nidation on the simulated endometria) revealed that at least two populations of cell types were recognisable as layers . This is interpreted as an early sign of morphogenesis and the first visible steps of differentiation . The presence of mitotic figures indicates viability and continuing growth . Electronmicroscopy of cell types grown under conditions simulating in vivo tissue architectonics showed overtly less cytopathology and better cell function . Simulated endometria may, therefore, serve as an attractive model for studying early mammalian embryogenesis and the effects of toxic agents. J Struct Funct Genomics, 2004, 5(1-2), 59 - 61 Cloning grills: high throughput cloning for structural genomics; Mehlin C et al.; Cloning grills are aluminum grids designed to divide an agar plate into segments, thereby multiplying the number of E . coli cultures which can be streaked out on a single plate . The grills are autoclaved and placed in square petri dishes immediately after hot agar is poured . When the agar solidifies, the grill remains embedded in the media, and each of the 12 lanes accommodates the streaking out of a single culture . As the spacing of the grill lanes is the same as that of a 96-well plate, 12 cultures can be streaked at a time using a 12-channel pipette . This allows a plate of 96 cultures to be rapidly and accurately plated for colony isolation on only eight agar plates. J Allergy Clin Immunol, 2004 Jun, 113(6), 1172 - 7 Airborne cat allergen reduction in classrooms that use special school clothing or ban pet ownership; Karlsson AS et al.; BACKGROUND: Allergens from furred animals are brought to school mainly via clothing of pet owners . Asthmatic children allergic to cat have more symptoms when attending a class with many cat owners, and some schools allocate specific resources to allergen avoidance measures . OBJECTIVE: The aim of the current study was to evaluate the effect of school clothing or pet owner-free classes compared with control classes on airborne cat allergen levels and to investigate attitudes and allergic symptoms among the children . METHODS: Allergen measurements were performed prospectively in 2 classes with school clothing, 1 class of children who were not pet owners, and 3 control classes during a 6-week period in 2 consecutive years . Portable pumps and petri dishes were used for collection of airborne cat allergen, and a roller was used for sampling on children's clothes . Cat allergen (Fel d 1) was analyzed with enzyme-linked immunoassay and immunostaining . Both years, questionnaires were administered to the children . RESULTS: We found 4-fold to 6-fold lower airborne cat allergen levels in intervention classes compared with control classes . Levels of cat allergen were 3-fold higher on clothing of cat owners than of children without cats in control classes . Pet ownership ban seemed less accepted than school clothing as an intervention measure . CONCLUSION: For the first time, it has been shown that levels of airborne cat allergen can be reduced by allergen avoidance measures at school by using school clothing or pet ownership ban, and that both measures are equally efficient . The clinical effect of these interventions remains to be evaluated. J Plant Physiol, 2004 May, 161(5), 509 - 17 Manipulation of the onset of ectomycorrhiza formation by indole-3-acetic acid, activated charcoal or relative humidity in the association between oak microcuttings and Piloderma croceum: influence on plant development and photosynthesis; Herrmann S et al.; A Petri dish system in which development of oak (Quercus robur L.) microcuttings is stimulated by the late stage ectomycorrhizal (EM) fungus Piloderma croceum J . Erikss . & Hjortst . in a long pre-symbiotic stage was optimised to allow synchronous, rhythmic plant growth . Addition of indole-3-acetic acid or activated charcoal to the medium caused an early and more intensive EM formation coupled with suppression of most developmental effects of P . croceum . Leaf area, chlorophyll fluorescence, and content were compared in inoculated and uninoculated plants grown at two relative humidity levels (45 and 95%) and under consideration of three possible answers to inoculation, that is, no or EM formation after the 1st or the 2nd shoot flush . The culture conditions for uninoculated plants were suboptimal, leading toward photochemical stress reflected by a non photochemical quenching (qE) increase and a reduced Chl content at the end of the assay . Prior to EM formation, inoculation itself enhanced the optimal (Fv/Fm) and effective (phiPSII) quantum yield in leaves of the 1st shoot flush under reduced relative humidity . It also fully protected the plants against stress during the complete assays . The results indicate that inoculated plants only form EM once they have acquired a sufficient development level and C-providing capacity . However, the fungus actively improves the development and photosynthesis of plants up to the pre-mycorrhizal stage, helping them to reach this capacity. J Biomed Mater Res, 2004 Jul 15, 70B(1), 43 - 8 Effects of laser-modified polystyrene substrate on CHO cell growth and alignment; Zhu B et al.; Biomaterial surface chemistry and nanoscale topography of biomaterials can significantly influence cell behavior in vitro . Polystyrene (PS) Petri dishes were subjected to Nd:YAG laser irradiation at 266 nm, which resulted in well-defined three-dimensional (3D) periodic nanoscale surface topographies and surface oxidation . The surface changes were analyzed by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and a contact-angle goniometer . The samples were then used to investigate the cell behavior of Chinese hamster ovary (CHO) cells . The surface laser modification affected the CHO cell adhesion and alignment, and caused morphological changes in comparison with unmodified PS . The results obtained from the cell-behavior studies revealed that nanoscale hydrophilic surface topography cues affected the adhesion, extension, alignment, and morphology of cells . Mycorrhiza, 2004 Jul, 14(3), 203 - 7 Epub 2003 Aug 05. Comparison of 233U and 33P uptake and translocation by the arbuscular mycorrhizal fungus Glomus intraradices in root organ culture conditions; Rufyikiri G et al.; This study aimed to quantify and compare 233U and 33P uptake and translocation by hyphae of the arbuscular mycorrhizal (AM) fungus Glomus intraradices in root organ culture conditions with transformed carrot (Daucus carota L.) roots as host . Mycorrhizal roots were grown in two-compartment Petri dishes to spatially separate a root compartment (RC) and a hyphal compartment (HC) . The HC was labelled with 8.33 Bq 233U ml(-1) and 13.33 Bq 33P ml(-1) . After 2 weeks contact between hyphae and the labelled solution, 233U and 33P activities were measured in the RC and in the HC . 233U and 33P were taken up by the extraradical AM mycelium grown in the HC and this uptake represented 4.4% and 16% of the initial isotope supply, respectively . The translocation into roots developing in the RC via hyphae accounted for 5.9% and 72% of the initial isotope supply, respectively . Thus, both uptake and translocation were much higher for 33P than for 233U . This suggests (1) the existence in hyphal tissues of efficient mechanisms limiting the uptake and translocation of non-essential elements such as U, and (2) that the hyphae have a higher sequestration than translocation function for U, and the converse for P. Appl Environ Microbiol, 2004 Jun, 70(6), 3512 - 20 Detection and quantification of airborne conidia of Fusarium circinatum, the causal agent of pine pitch canker, from two California sites by using a real-time PCR approach combined with a simple spore trapping method; Schweigkofler W et al.; Pinus radiata (Monterey pine), a tree native to coastal California and Mexico, is widely planted worldwide for timber production . A major threat to Monterey pine plantations is the fungal disease pine pitch canker, caused by Fusarium circinatum (Hypocreales) . We present a novel trapping approach using filter paper in combination with a rapid molecular method to detect the presence of inoculum in the air . The assay is also useful for diagnosing the presence of the pathogen on plants . The test is based on the F . circinatum specific primer pair CIRC1A-CIRC4A, which amplifies a 360-bp DNA fragment in the intergenic spacer region of the nuclear ribosomal operon . Real-time PCR was used to calculate the number of fungal spores present in each reaction mixture by comparing the threshold cycle (Ct) of unknown spore samples to the Ct values of standards with known amounts of F . circinatum spores . The filter paper method allows prolonged and more sensitive spore sampling in the field compared to traditional traps using petri dishes filled with selective medium . A field test at two sites in coastal California infested with pine pitch canker was carried out during the summer and fall of 2002 . Spore counts were in the range of ca . 1 x 10(3) to ca . 7 x 10(5)/m(2), with the highest spore counts in the fall, suggesting a seasonal fluctuation. J Agric Food Chem, 2004 Jun 2, 52(11), 3389 - 93 Effect of heat treatment on the antioxidant activity of extracts from citrus peels; Jeong SM et al.; The effect of heat treatment on the antioxidant activity of extracts from Citrus unshiu peels was evaluated . Citrus peels (CP) (5 g) were placed in Pyrex Petri dishes (8.0 cm diameter) and heat-treated at 50, 100, or 150 degrees C for 10, 20, 30, 40, 50, and 60 min in an electric muffle furnace . After heat treatment, 70% ethanol extract (EE) and water extract (WE) (0.1 g/10 mL) of CP were prepared, and total phenol contents (TPC), radical scavenging activity (RSA), and reducing power of the extracts were determined . The antioxidant activities of CP extracts increased as heating temperature increased . For example, heat treatment of CP at 150 degrees C for 60 min increased the TPC, RSA, and reducing power of EE from 71.8 to 171.0 microM, from 29.64 to 64.25%, and from 0.45 to 0.82, respectively, compared to non-heat-treated control . In the case of WE from CP heat-treated at the same conditions (150 degrees C for 60 min), the TPC, RSA, and reducing power also increased from 84.4 to 204.9 microM, from 15.81 to 58.26%, and from 0.27 to 0.96, respectively . Several low molecular weight phenolic compounds such as 2,3-diacetyl-1-phenylnaphthalene, ferulic acid, p-hydroxybenzaldoxime, 5-hydroxyvaleric acid, 2,3-diacetyl-1-phenylnaphthalene, and vanillic acid were newly formed in the CP heated at 150 degrees C for 30 min . These results indicated that the antioxidant activity of CP extracts was significantly affected by heating temperature and duration of treatment on CP and that the heating process can be used as a tool for increasing the antioxidant activity of CP. Commun Agric Appl Biol Sci, 2003, 68(4 Pt B), 711 - 5 Activity of some chemicals in the control of Botrytis cinerea on roses; Wojdyla AT; In the control of grey mould (Botrytis cinerea) on rose petals all 15 tested fungicides used as a spray, suppressed very effectively development of necrosis . But the best results in suppression of necrosis development, more than 90% effectiveness gave: Amistar 250 SC (azoxystrobin), Bravo 500 SC (chlorothalonil), Discus 500 WG (kresoxim methyl), Euparen 50 WP (dichlofluanid) Euparen Multi 50 WG (tolyfluanid), Folpan 80 WG (folpet), Kaptan zaw . 50 WP (captan), Penncozeb 80 WP (mancozeb), Ronilan 50 WP (vinclozolin), Rovral Flo 255 SC (iprudione), Sumilex 500 SC (procymidone) and Teldor 500 SC (fenhexamid) . When Dithane M-45 80 WP (mancozeb), Sarbrawit 530 SC (chlorothalonil + carbendazim) or Topsin M 70 WP (thiophanate methyl) were used, the diameter of spots was about 6-times smaller than on control flowers (about 84% effectiveness) . All tested fungicides used for spraying of flower petals with abundant visible sporulation of B . cinerea were ineffective in the inhibition of spore germination . But when Petri dishes with potato-dextrose-agar were sprayed with tested fungicides and inoculated with spore suspension, Euparen 50 WP, Euparen Multi 50 WG, Folpan 80 WG, Kaptan zaw . 50 WP and Penncozeb 80 WP completely suppressed conidia germination. Commun Agric Appl Biol Sci, 2003, 68(4 Pt B), 599 - 607 Pathogenicity of Phytophthora infestans on solanaceous and asteraceous plant species in Cameroon; Fontem DA et al.; Experiments were conducted to detect potential hosts of Phytophthora infestans, causal agent of potato late blight . Isolates of P . infestans recovered from garden huckleberry (Solanum scabrum), potato (S . tuberosum) and tomato (Lycopersicon esculentum) were inoculated on detached leaves of 12 solanaceous and 14 asteraceous plant species collected in and around potato fields in the western highlands of Cameroon . The inoculated leaves were maintained in Petri dishes in the laboratory for 7 days at 20-22 degrees C . Isolates of P . infestans from huckleberry, potato and tomato infected the same host plants as well as the gboma eggplant (Solanum macrocarpon) and two asteraceous weeds, Ageratum conyzoides and Solanecio biafrae . Inocula recovered from infected S . macrocarpon caused late blight symptoms on huckleberry, potato and tomato both under laboratory and screen house conditions, while those from both asteraceous weeds reinfected potato in the laboratory and inoculum from S . biafrae reinfected huckleberry in the screen house . This is a first observation of late blight infection on S . macrocarpon, A . conyzoides and S . biafrae in Cameroon . Results indicate the importance of both asteraceous weeds and S . macrocarpon as potential alternative hosts for P . infestans in garden huckleberry, potato or tomato fields. Commun Agric Appl Biol Sci, 2003, 68(4 Pt A), 397 - 402 Cross-resistance to ACCase inhibitors of Lolium multiflorum, Lolium perenne and Lolium rigidum found in Chile; Michitte P et al.; APPs (aryloxyphenoxypropionates) and CHDs (cyclohexanediones) are two of the most important groups used post-emergence for the control of grass weeds . They inhibit the lipid synthesis in plants by interfering with the activity of the enzyme Acetyl-coenzyme A carboxylase (ACCase), acting at a meristematic level . The resistance patterns of the biotypes characterized seem to indicate the existence of different degrees of resistance . It is thus possible to identify biotypes presenting cross-resistance only to certain APPs, to APPs and CHDs, or only to CHDs . The objective of this work was to evaluate the cross-resistance to fenoxaprop-P-ethyl, fluazifop-P-butyl, propaquizafop, cyhalofop-butyl, haloxyfop-R-methyl, tralkoxydim and tepraloxydim in three species of Lolium (L . multiflorum, L . perenne, and L . rigidum) resistant to diclofop-methyl . The assays were conducted with petri-dishes in which, over increasing doses, fifty seeds per biotype and dose were located in each dish . Two weeks later, the following parameters were evaluated: germination (%), number of roots, radicle length, plumule length, and fresh weight reduction (%) . Based on plumule length and fresh weight reduction (%), diclofop-methyl resistant biotypes showed cross-resistance to fenoxaprop-P-ethyl, fluazifop-P-butyl, cyhalofop-butyl, haloxyfop-R-methyl, but not to propaquizafop, tralkoxydim and tepraloxydim . The parameters germination (%), number of roots or root length did not show a good relation between the dose and its efficacy (curves of dose response) for any of the susceptible and resistant biotypes studied. Commun Agric Appl Biol Sci, 2003, 68(4 Pt A), 331 - 4 Is it possible to detect Echinochloa spp . tolerance to ACCase-inhibiting herbicides using a simple quick tolerance test? Ruiz-Santaella JP, Bakkali Y, Fischer AJ, De Prado R. The objective of this work was to evaluate the reliability of one quick tolerance test that would enable us, in only six days, to quantify tolerance levels of one biotype to one or more herbicides . For this purpose, we evaluated tolerance levels to cyhalofop-butyl of five Echinochloa spp . biotypes: muricata, crus-galli, crus-pavonis, oryzicola and utilis, with unknown records of herbicide treatment . Moreover, two biotypes of Echinochloa phyllopogon were tested, one of them resistant (R) to fenoxaprop-P-ethyl and the other one susceptible (S) . The Oryza sativa Puntal var . and Oryza sativa var . (wild rice) were also checked in order to demonstrate cyhalofop-butyl selectivity profiles . The assays were conducted with petri-dishes where, over the following increasing doses, fifty seeds per biotype and dose, were placed in each one: 0, 3, 6, 12, 18, 24, 48 and 60 ppm of cyhalofop-butyl and codacide oil adjuvant . Six days later, the following parameters were evaluated: plumule length, root length, germination (%) and fresh weight reduction (%) . E . muricata germination was reduced by 50% with a 6 ppm dose . A non linear relation dose-germination was observed in the rest of the biotypes . The plumule length permitted the ranking of the biotypes into three groups: (1) resistant (E . phyllopogon R) or very tolerant (Oryza sativa Puntal var . and wild rice), those with a reduction of less than 30%, (2) tolerant, between 30 and 60% (E . oryzicola), and (3) susceptible, with a reduction of over 80% (E . muricata, E . crus-galli, E . crus-pavonis, E . utilis and E . phyllopogon S) . Only Oryzo sativa and E . phyllopogon R had a minimal reduction of fresh weight (less than 15%) . The assays were repeated with whole plants and we checked the accuracy of this test that indicated the resistance level of one biotype in a quick, reliable and economic way. Allergy, 2004 Jun, 59(6), 661 - 7 Allergen avoidance does not alter airborne cat allergen levels in classrooms; Karlsson AS et al.; BACKGROUND: Some schools in Sweden offer allergen avoidance classrooms for allergic children with severe asthma . However, the measures commonly used to achieve a reduction in allergen levels have not been properly evaluated . The aim of the present prospective study was to study whether the levels of airborne cat allergen are altered after introducing feasible intervention measures in classrooms, without interfering with peoples' freedom of choice regarding pet ownership . METHODS: Twenty-five classes, including five established allergy prevention classrooms participated in the study during a school year . After one term, six classes underwent a number of intervention measures recommended by the Swedish National Institute of Public Health . Curtains, upholstery and plants were removed, bookshelves were replaced with cupboards and regular cleaning was increased . Airborne dust was collected weekly (32 weeks) using duplicate Petri dishes (n = 1574) and on six occasions using two personal air samplers in each class (n = 264) . RESULTS: Airborne cat allergen levels were showing a similar variability throughout the whole study in all classes . Despite extensive measures in order to reduce allergen exposure, cat allergen levels were unaltered in the six classes after intervention . Allergen levels were not significantly lower in the established allergy prevention classes, compared with the other classes . Cat allergen levels differed, however, significantly between classes with few and many cat owners (P < 0.05) . CONCLUSIONS: We conclude that the recommended allergen avoidance measures used in this study did not reduce airborne cat allergen . It seems plausible that measures that fail to reduce allergen levels also fail to influence health status in allergic children but this remains to be shown. J Environ Sci Health B, 2004 Mar, 39(2), 297 - 309 A study on the environmental degradation of pesticides azinphos methyl and parathion methyl; Athanasopoulos PE et al.; The effect of environmental parameters (temperature and relative humidity) on the degradation rate of azinphos methyl and parathion methyl was studied . Proprietary emulsifiable concentrates were diluted and added to each of 90 glass Petri dishes for each pesticide and were left overnight to dry . Petri dishes were placed in 18 air-tight containers (9 for each pesticide) in which were created environments with relative humidity (RH) of 60, 82, and 96% . The containers were stored at 0, 20, and 40 degrees C . From the experimental results best fit curves, kinetic equations, rate constants, and half-lives were calculated . Half-lives of azinphos methyl for the RH studied were, from 124 to 267 days at 0 degrees C, from 89 to 231 days at 20 degrees C, and from 25 to 71 days at 40 degrees C . Corresponding half-lives for parathion methyl were from 48 to 57 days at 0 degrees C, from 9.2 to 10.5 days at 20 degrees C and from 1.3 to 1.5 days at 40 degrees C . The results were correlated with relevant results from the decomposition of the same or similar pesticides on apples both, on the trees and during refrigerated storage . These correlations are suggesting that biological factors strongly affected the decomposition rate of azinphos methyl . On the contrary the decomposition of parathion methyl was mainly affected by environmental rather than biological factors. Biomaterials, 2004 Aug, 25(19), 4741 - 8 Surface properties and biocompatibility of solvent-cast poly{-caprolactone} films; Tang ZG et al.; Poly(epsilon-caprolactone) (PCL) was dissolved in four solvent systems, chloroform, tetrahydrofuran, acetone and ethyl acetate, and cast onto glass Petri dishes . The surface properties of the resulting films were investigated . The extent to which their properties were determined by the solvent used in each case was quantified in terms of contact angle, surface morphology, attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR), and the adhesion and proliferation of fibroblasts by direct contact . The surface of the PCL film in contact with glass was denoted the SG surface, and the other, which was exposed to the gas phase, a mixture of air and residual solvent vapour, was denoted the SA surface . In the case of hydrophobic solvent systems, the advancing contact angle of the SG surface was always lower than that of the SA surface . With hydrophilic solvent systems, on the other hand, the advancing contact angle of the SG film surface was higher when the contact angle of the Petri dish was higher than that of the gaseous mixture of the air and solvent vapour, otherwise it was lower or equal to that of the surface on which it was cast . The surface morphology was dictated by the solubility of PCL in the respective solvent systems: high dissolution solvents such as chloroform and tetrahydrofuran produced films that comprised PCL aggregates, the particles being larger in the case of chloroform, whereas the less efficient solvents (acetone and ethyl acetate) resulted in a filamentous structure . The ATR-FTIR results confirmed that the chemistry of the SA surfaces differed according to the solvent system used . Preliminary cell culture experiments carried out with the PCL films established that murine (L929) fibroblasts grew well on all surfaces regardless of the solvent used, although the rates of adhesion and proliferation were not as great as on tissue culture plastic controls . Of all the surfaces examined in this study, the cells favoured the SG aspect of ethyl acetate cast PCL films, the surface of which had the finest pore size and relatively low contact angle. Can J Microbiol, 2004 Mar, 50(3), 221 - 4 Disinfection of selected Aspergillus spp . using ultraviolet germicidal irradiation; Green CF et al.; AIMS: The efficacy of ultraviolet germicidal irradiation (UVGI) and the UVGI dose necessary to inactivate fungal spores on an agar surface for cultures of Aspergillus flavus and Aspergillus fumigatus were determined . METHODS AND RESULTS: A four-chambered UVGI testing unit with a 9-W, Phillips, low pressure, mercury UVGI lamp in each chamber was used in this study . An aperture was adjusted to provide 50, 100, 150, and 200 micro W/cm2 of uniform flux to the surfaces of the Petri dish, resulting in a total UVGI dose to the surface of the Petri dishes ranging from 12 to 96 mJ/cm2 . The UVGI dose necessary to inactivate 90% of the A . flavus and A . fumigatus was 35 and 54 mJ/cm2, respectively . CONCLUSIONS: UVGI can be used to inactivate culturable fungal spores . Aspergillus flavus was more susceptible than A . fumigatus to UVGI . SIGNIFICANCE AND IMPACT OF THE STUDY: These results may not be directly correlated to the effect of UVGI on airborne fungal spores, but they indicate that current technology may not be efficacious as a supplement to ventilation unless it can provide higher doses of UVGI to kill spores traveling through the irradiated zone. Phys Med Biol, 2004 Mar 21, 49(6), N83 - 92 Novel high-resolution temperature probe for radiofrequency dosimetry; Schuderer J et al.; A novel integrated thermistor probe for temperature evaluations in radiofrequency-heated environments was realized . The probe's sensitive area is based on a highly resistive 50 microm x 100 microm layer of amorphous germanium processed on a glass tip . The small dimensions allow measurements with a distance as close as 150 microm from solid boundaries . Due to its high temperature resolution of 4 mK and its short response time of the order of 10 ms, the sensor is very well suited for dosimetric measurements in strong absorption gradients . The influence of radiofrequency (RF) electric fields on the signal is minimized due to the high resistance of the sensor and the leads . The probe was successfully used to determine the highly nonuniform absorption distribution resulting from the RF exposure of cell cultures placed in Petri dishes. Neuroscience, 2004, 125(3), 591 - 604 Alignment of glial cells stimulates directional neurite growth of CNS neurons in vitro; Deumens R et al.; Olfactory ensheathing cells (OECs) together with olfactory nerve fibroblasts (ONFs) and neonatal astrocytes are potent stimulators of neurite growth in adulthood and during development, respectively . Since it is known that alignment of glial cells is important for the correct outgrowth of axon tracts, it was hypothesized that the alignment of glial cells stimulates directional and enhanced neurite outgrowth . Adult OEC/ONF and neonatal astrocytes were cultured either on biodegradable poly(d,l)-lactide matrices or in Petri dishes for 4 days . Thereafter neonatal cerebral cortical neurons were added . After a 2-days coculture period the cultures were fixed and processed for a combined MAP-2 and phosphorylated neurofilament (RT97) staining . The neurite growth (neurite elongation and neurite formation) and the neurite direction were assessed . We show that (1) . OEC/ONF cultures are more potent in stimulating the length of the longest neurite of cocultured neurons, (2) . alignment of glial is achieved in vitro on our biomatrices, (3) . aligned glial/biomatrix complexes do not enhance neurite growth, and (4) . aligned glial/biomatrix complexes direct neurite outgrowth . These data have significant implications for in vivo experiments focusing on glial transplantation . Transplanting glial/biomatrix complexes may stimulate the directional regrowth of severed axons across a lesion site. Lasers Surg Med, 2004, 34(4), 323 - 8 Effect of low-power He-Ne laser irradiation on rabbit articular chondrocytes in vitro; Jia YL et al.; BACKGROUND AND OBJECTIVES: In the orthopaedic field, the repair of articular cartilage is still a difficult problem, because of the physiological characters of cartilaginous tissues and chondrocytes . To find an effective method of stimulating their regeneration, this in vitro study focuses on the biostimulation of rabbit articular chondrocytes by low-power He-Ne laser . STUDY DESIGN/MATERIALS AND METHODS: The articular chondrocytes isolated from the cartilage of the medial condyle of the femur of the rabbit were incubated in DMEM/HamF(12) medium . The second passage culture were spread on 24 petri dishes and were irradiated with laser at power output of 2-12 mW for 6.5 minutes, corresponding to the energy density of 1-6 J/cm(2) . Laser treatment was performed three times at a 24-hour interval . After lasering, incubation was continued for 24 hours . Non-irradiated cells were kept under the same conditions as the irradiated ones . The cell proliferation activity was evaluated with a XTT colorimetric method and the cell secretion activity was analyzed by metachromasia and immunocytochemistry . RESULTS: Irradiation of 4-6 J/cm(2) increased the cell numbers and revealed a considerably higher cell proliferation activity comparing to control cultures . Thereinto, the energy density of 4 and 5 J/cm(2) remarkably increased cell growth, with positive effect on synthesis and secretion of extracellular matrix . CONCLUSIONS: The present study showed that a particular laser irradiation stimulates articular chondrocytes proliferation and secretion . These findings might be clinically relevant, indicating that low-power laser irradiation treatment is likely to achieve the repair of articular cartilage in clinic . Invest Radiol, 2004 Mar, 39(3), 187 - 95 Detection of individual microbubbles of ultrasound contrast agents: imaging of free-floating and targeted bubbles; Klibanov AL et al.; RATIONALE AND OBJECTIVES: During echo examinations with microbubble contrast, individual "dots" of ultrasound reflection can be visualized . To address the question whether these signals represent individual microbubbles, very dilute suspensions of ultrasound contrast agents or individual microbubbles attached to Petri dishes were prepared and studied by ultrasound imaging . METHODS: Microbubble suspensions were diluted in saline and evaluated by a clinical ultrasound imaging system . Microbubble concentration was verified by Coulter counter . Single microbubble preparation on a Petri dish was established by streptavidin-biotin interaction under microscopy control and subjected to ultrasound imaging . RESULTS: Ultrasound of dilute microbubble dispersions demonstrated distinct white foci; concentration of these sites was consistent with signals from individual microbubbles as determined by Coulter . Individual microbubbles immobilized on polystyrene were also visualized by ultrasound . CONCLUSION: Ultrasound medical systems can resolve backscatter signals from individual microbubbles of ultrasound contrast, both in solution and in the targeted immobilized state, implying picogram sensitivity. J Med Entomol, 2004 Mar, 41(2), 249 - 54 Comparative activity of deet and AI3-37220 repellents against the ticks Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae) in laboratory bioassays; Carroll JF et al.; The repellents N,N-diethyl-3-methylbenzamide (deet) and racemic 2-methylpiperidinyl-3-cyclohexene-1-carboxamide (AI3-37220) were evaluated using two different laboratory bioassays to determine their relative effectiveness against host-seeking nymphs of the blacklegged tick, Ixodes scapularis Say, and the lone star tick, Amblyomma americanum (L.) . In a petri dish bioassay, ticks were released within a ring of repellent on a horizontal filter paper disk . In the second bioassay, ticks were allowed to climb a vertical strip of filter paper whose central portion was treated with a repellent . Deet and AI3-37220 were more effective against I . scapularis than A . americanum nymphs . In the petri dish bioassay, none of the concentrations of deet or AI3-37220 tested confined A . americanum within the treated ring . However, in the vertical bioassay, both species exhibited avoidance of the repellents, and I . scapularis was repelled by much lower concentrations than A . americanum . I . scapularis were repelled by lower concentrations in the vertical bioassay than in the petri dish bioassay . Deet was slightly more effective against I . scapularis than AI3-37220 in both bioassays, but AI3-37220 was significantly more effective than deet against A . americanum in the vertical bioassay. Fungal Genet Biol, 2004 May, 41(5), 563 - 70 Nuclear reassortment between vegetative mycelia in natural populations of the basidiomycete Heterobasidion annosum; Johannesson H et al.; Somatic incompatibility is not an absolute block to nuclear exchange between incompatible mycelia of the basidiomycete Heterobasidion annosum in vitro . Under laboratory conditions new heterokaryotic genotypes can be isolated from the gap between incompatible heterokaryons, and nuclear migration between pairs of heterokaryons grown in Petri dishes has been observed . In this study, we test the hypothesis of nuclear transfer and reassortment between heterokaryotic mycelia in natural populations of H . annosum . We developed six microsatellite markers to genotype nuclei populating 21 somatically incompatible mycelia of H . annosum isolated from a single stump of Picea abies, and found that the detected heterokaryons share nuclei; 10 of the nuclear haplotypes were found in more than one mycelium . In one isolate, four nuclear types were found in the same mycelium . These findings indicate that new heterokaryons can be formed as a result of nuclear reassortment between incompatible heterokaryotic mycelia in nature. Cryo Letters, 2004 Jan-Feb, 25(1), 51 - 8 Cryopreservation of in vitro-grown shoot tips of cassava by encapsulation-vitrification method; Charoensub R et al.; Shoot tips of cassava (Manihot esculenta Crantz) in vitro plantlets were successfully cryopreserved using the encapsulation-vitrification technique . Nodal cuttings of 5 mm length with one leaf were cultured on modified MS medium in Petri dishes (90 mm x 20 mm) for about 28 days . Excised shoot tips were precultured on sucrose enriched (0.3 M) medium for 16 h, encapsulated and osmoprotected with a mixture of 2 M glycerol and 0.6 M sucrose for 90 min at 25 degree c before dehydration with PVS2 at 0 degree C for 4 h, then plunged in liquid nitrogen . Successfully vitrified shoot tips resumed growth within 3 days, without intermediary callus formation, and developed shoots . Shoot tips sampled from 21 day-old plantlets produced the highest survival of 80 % . The percentage survival of vitrified shoot tips differed from 38 to 80 % depending on the day of excision . The protocol was successfully applied to four cultivars of cassava with about 80 % average percentage of survival. Biochim Biophys Acta, 2004 Mar 17, 1671(1-3), 18 - 25 A simple model for the study of effects of the extracellular matrix on the cell morphology in vitro; Kuz'minykh EV et al.; In the present work, a simple technique is proposed to study the effects of native extracellular matrix (ECM) of one cell type on the properties of other cell types . It is based on a procedure in which, after cells of one type are removed from the substrate, cells of another type are seeded on the same substrate . To obtain preparations of native ECM, cells were removed from the substrate by 0.02% EDTA only, without any proteolytic enzymes . Cells were placed on coverslips in standard Petri dishes and incubated in a culture medium for a time sufficient for adhesion and spreading, but not long enough to undergo mitosis . Up to four coverslips per Petri dish can be incubated, and various combinations of ECM and cell types can be used in one dish . It is important, therefore, that the different "ECM-cell" combinations are present in the same culture medium . For evaluation of ECM effects, the area occupied by the cell on a substrate and the perimeter of the cell were measured, and frequencies of cell distribution were calculated according to these parameters. J Altern Complement Med, 2004 Feb, 10(1), 113 - 22 Measuring effects of music, noise, and healing energy using a seed germination bioassay; Creath K et al.; OBJECTIVE: To measure biologic effects of music, noise, and healing energy without human preferences or placebo effects using seed germination as an objective biomarker . METHODS: A series of five experiments were performed utilizing okra and zucchini seeds germinated in acoustically shielded, thermally insulated, dark, humid growth chambers . Conditions compared were an untreated control, musical sound, pink noise, and healing energy . Healing energy was administered for 15-20 minutes every 12 hours with the intention that the treated seeds would germinate faster than the untreated seeds . The objective marker was the number of seeds sprouted out of groups of 25 seeds counted at 12-hour intervals over a 72-hour growing period . Temperature and relative humidity were monitored every 15 minutes inside the seed germination containers . A total of 14 trials were run testing a total of 4600 seeds . RESULTS: Musical sound had a highly statistically significant effect on the number of seeds sprouted compared to the untreated control over all five experiments for the main condition (p < 0.002) and over time (p < 0.000002) . This effect was independent of temperature, seed type, position in room, specific petri dish, and person doing the scoring . Musical sound had a significant effect compared to noise and an untreated control as a function of time (p < 0.03) while there was no significant difference between seeds exposed to noise and an untreated control . Healing energy also had a significant effect compared to an untreated control (main condition, p < 0.0006) and over time (p < 0.0001) with a magnitude of effect comparable to that of musical sound . CONCLUSION: This study suggests that sound vibrations (music and noise) as well as biofields (bioelectromagnetic and healing intention) both directly affect living biologic systems, and that a seed germination bioassay has the sensitivity to enable detection of effects caused by various applied energetic conditions. Environ Int, 2004 May, 30(3), 375 - 81 Assessment of application-rate dependent effects of a long-term fire retardant chemical (Fire Trol 934) on Typha domingensis germination; Angeler DG et al.; Although long-term fire retardants (LTR) gain increasingly acceptance as effective tools for wildfire management, recent studies indicate their potential harmfulness in the environment, especially in aquatic ecosystems . This study comprises a first laboratory impact assessment of LTR action in temporal Mediterranean wetlands, using seed germination of Typha domingensis as indicator of impact . Our aim was to identify application rates upon which seed germination could be significantly affected . We tested for low (1 l m(-2)) and high (3 l m(-2)) application rates of Fire Trol 934 which are recommended by the manufacturers as a function of fuel characteristics . In addition, we simulated the impact of a higher application rate of 5 l m(-2) because inhomogeneous dispersal of the LTR during fire control and prevention operations can result in locally elevated applications . Results of a microcosm experiment indicate that application rates of 1 or 3 l m(-2) can impact Typha germination rates in the short-term via indirect LTR-mediated effects on water quality, which suppressed necessary cues for germination . However, a subsequent experiment with Petri dishes, using seeds isolated from the LTR treated sediments did not show significantly different germination rates between the control and the treatments with application rates of 1 or 3 l m(-2) . This suggests that retardant pre-application germination success could be recovered in nature once the retardant is eliminated . By contrast, seeds almost completely failed to germinate in the microcosm experiment and the subsequent Petri dish essay when an application rate of 5 l m(-2) was used . This suggests a critical level upon which Typha seed germination may be perpetually limited . Research should be extended to other plant species to provide fire managers with guidelines for environmentally safe use of LTR in the Mediterranean region. Appl Radiat Isot, 2004 Feb-Apr, 60(2-4), 227 - 32 A hybrid method to compute accurate efficiencies for volume samples in gamma-ray spectrometry; Garcia-Talavera M et al.; In recent years, Monte Carlo (MC) methods have been increasingly applied to cope with variability in photopeak efficiencies due to matrix effects . But to obtain proper results only by numerical simulation, especially at low energies, sample bulk density and chemical composition must be well characterized . In this paper, we propose a method that combines both experimental measurements and MC simulations, being applicable to matrices of unknown composition . A transmission measurement of a 210Pb point source through the sample allows one to compute accurately its photopeak efficiencies at energies above 46.5 keV . The method is validated for several inorganic and organic matrices measured in Petri dishes geometry. Tree Physiol, 1987 Dec, 3(4), 375 - 86 Ecophysiological characteristics of the seed of the tropical forest pioneer Urera caracasana (Urticaceae); Orozco-Segovia A et al.; Urera caracasana (Jacq.) Griseb is a small, fast-growing evergreen pioneer tree which colonizes openings in the tropical rain forest of 'Los Tuxtlas', Veracruz, Mexico . Annual seed production by 10 trees was estimated to range from 0.4 x 10(5) to 1.6 x 10(6) seeds per tree . Fifteen species of resident and migratory birds were observed to visit the plants and disperse the seeds . Most seeds imbibed in petri dishes placed on the forest floor beneath a small opening in the canopy, where the red/far-red ratio of the light was around 1.0, germinated within 2 weeks . Germination beneath the forest canopy, where the red/far-red ratio of the light was approximately 0.2, was much slower and did not exceed 40% . A minimum of 4 h daily exposure to unfiltered natural light was required for rapid germination in the forest . In the laboratory rapid germination at 25 degrees C required a minimum of 4 h white light . However, if temperature during the light period was 35 degrees C, 30 min exposure to white light daily was sufficient to induce germination . Seeds that failed to germinate when imbibed for 60-360 days in petri dishes placed on the forest floor beneath the forest canopy, germinated rapidly in the laboratory when held at 25 degrees C and exposed daily to 12 h white or red light . Seeds imbibed for 120 days or more beneath the forest canopy were also induced to germinate by 12 h daily exposure to far-red light when this was combined with a 35/25 degrees C day/night temperature regime . Seeds buried in vermiculite-filled nylon mesh bags disappeared rapidly with few remaining after 2 months . Seeds that survived remained viable and germinated when incubated in petri dishes at 25 degrees C . Initially, buried seeds required light for germination . However, after 17 months' burial, seeds germinated in darkness when transferred to the laboratory and incubated on agar at 25 degrees C. Exp Appl Acarol, 2003, 31(1-2), 1 - 14 Propensity towards cannibalism among Hypoaspis aculeifer and H . miles, two soil-dwelling predatory mite species; Berndt O et al.; In biological control programmes, the two predatory soil mites Hypoaspis aculeifer and H . miles are often applied against soil-borne pests like mushroom flies, springtails and mites . Although the mites show high consumption rates on varying prey types in Petri dish experiments and in greenhouses, their overall efficiency is sometimes limited . We hypothesized that intraspecific interactions, like cannibalism, could contribute to this decreased competence . Therefore, experiments were conducted to show the propensity of H . aculeifer and H . miles to cannibalise . Adult mites and nymphs were introduced as predators with conspecific eggs, larvae, nymphs, adult females or males as prey and the number of killed individuals was recorded . Additionally, the oviposition rate on conspecific prey was quantified and the correlation with the number of prey consumed was calculated to assess the influence of cannibalism on egg production . The results illustrate that cannibalism occurs infrequently in both Hypoaspis spp., the only exception being H . aculeifer nymphs, which cannibalised one conspecific egg per day . Moreover, cannibalism never occurred in the presence of alternative prey . Oviposition rate decreased during the experiment in both species but it was positively correlated with the cannibalism rate only for H . aculeifer . The benefit of cannibalism for populations of H . aculeifer and H . miles is discussed. Scand J Surg, 2003, 92(4), 241 - 7 Tissue engineering--body parts from the Petri dish; Kratz G et al.; The development of methods to regenerate human tissues and organs by tissue engineering (TE), will have a dramatic influence on many medical specialities in the future . The essence of plastic surgery is to reconstruct disrupted and damaged tissues by the use of a plethora of techniques spanning from small local skin flaps to highly advanced microsurgery and free composite grafts . However, these methods only focus on moving tissue from one part of the patient to another without actual regeneration . To be able to take the next step in development of the speciality it is of necessity to address this issue . Hence it follows naturally that plastic surgeons lead and represent the driving force of the development within the research of tissue engineering . In this paper we would like to present active research and also give an overview of areas in tissue engineering that are of special interest to the plastic surgeon. J Photochem Photobiol B, 2004 Jan 23, 73(1-2), 59 - 66 Alternative methods to evaluate the protective ability of sunscreen against photo-genotoxicity; Toyoshima M et al.; Numerous epidemiological investigations show that sunlight is carcinogenic to humans and that the use of sunscreen may be effective in decreasing the risk of skin cancer . The biological activity of a sunscreen is evaluated by its ability to protect human skin from erythema as represented by a Sun Protection Factor (SPF) . We propose that the sunscreen's protective effect against sunlight-induced genotoxicity, including mutation, should also be taken into account . In this study we examined the protective ability of sunscreens against natural sunlight and UV-induced genotoxicity in Drosophila somatic cells . We prepared three kinds of sunscreen samples, each with an SPF value of 20, 40 or 60 and compared their protective activities with commercial sunscreens . When a sunscreen of SPF 20, 40 or 60 was pasted on the plastic cover of a petri dish in which Drosophila larvae were exposed to the sun or UV lamps, genotoxicity decreased as the SPF of the sunscreen increased, relative to levels of genotoxicity observed in samples without sunscreen . However, the protective abilities of sunscreens were unexpectedly not so different from each other . To reveal the relationship between the protective activity of sunscreen and the wavelength of light with which larvae were irradiated through the sunscreen, we measured the transmittance of light through the petri dish cover on which the sunscreen was pasted . Effective protection was demonstrated by removing components of light whose wavelengths were below 315 nm . We suggest, that the measurement of anti-genotoxic activity and the determination of the wavelengths of light transmitted through the sunscreen should be an alternative method for evaluating the effectiveness of a sunscreen. Bull Entomol Res, 2003 Dec, 93(6), 499 - 505 Intraguild predation among ladybeetles and a green lacewing: do the larval spines of Curinus coeruleus(Coleoptera: Coccinellidae) serve a defensive function? Michaud JP, Grant AK. Laboratory experiments examined interspecific interactions between larvae of three coccinellid species, Curinus coeruleus Mulsant (Chilocorinae), Harmonia axyridis Pallas and Olla v-nigrum (Mulsant) (Coccinellinae), and between these and larvae of the green lacewing, Chrysoperla rufilabris (Burmeister) . Larvae of C . coeruleus, although defended on their dorsal surface with long spines, had the smallest mandibles, were the slowest-moving, and the least successful in interspecific larval combat . The long spines of third instar C . coeruleusappeared to reduce their palatability as food to H . axyridis and O . v-nigrum larvae in choice tests with dead larvae, but were not an effective defence against these species in Petri dish arenas . Larvae of O . v-nigrum had a smooth dorsal surface, were intermediate in terms of mandible size, but were the fastest moving, a trait that benefited their survival in intraguild combat . Larvae of H . axyridis were intermediate with respect to dorsal spines and speed of movement, but had the largest mandibles . This species was the most effective intraguild combatant among the coccinellids and the only one to successfully compete against C . rufilabris larvae of similar age . The speed, manoeuverability and long mandibles of C . rufilabris enabled them to impale coccinellid larvae at a relatively safe distance . The spines of C . coeruleus larvae impeded laterally oriented attacks by C . rufilabris, but did not provide sustained protection from repeated attacks . Success in these interactions appeared largely a function of offensive weaponry (mandible size and morphology) and speed of movement, although the role of dorsal spines as defensive structures was not ruled out . Rates of larval cannibalism were highest for C . rufilabris and largely mirrored the level of aggression observed in interspecific combat for each species. J Chem Ecol, 2003 Nov, 29(11), 2413 - 24 Physiological assessment and path coefficient analysis to improve evaluation of alfalfa autotoxicity; Chon SU et al.; Reseeding of alfalfa is affected until autotoxic chemicals break down or are dispersed, often requiring a year or more . Bioassays of seed germination and early seedling growth, on agar medium in petri dishes, were conducted to evaluate autotoxic responses of 20 alfalfa germplasms to water-soluble extracts of alfalfa leaf tissue . Root length, 120 hr after placing imbibed seed on agar, was more sensitive to the autotoxin(s) than was hypocotyl length, germination speed, and final germination percentage . Path coefficient analyses showed variation in root length had 7-17 times more effect than variation in hypocotyl length in determining autotoxic effects on total seedling length . Although variations in seed size and germination rate were negatively associated (P < 0.05) with final root length, the autotoxin had little effect on these factors relative to that on root length . Germplasms in the control differed (P < 0.05) in root length, requiring tolerance to be evaluated as percent of control . Germplasms, as percent of control, differed significantly (P < 0.05) at extract concentrations of 1.0 and 4.0 g l(-1), but the range and LSD were more favorable for selection at 1.0 g l(-1) . Root length is appropriate for genetic assessments of tolerance to the autotoxin when expressed as percent of control. Mycopathologia, 2003, 156(4), 375 - 82 Pathogenicity of Metarhizium anisopliae (Metsch.) Sorokin and Beauveria bassiana (Balsamo) Vuillemin, to three adult fruit fly species: Ceratitis capitata (Weidemann), C . rosa var . fasciventris Karsch and C . cosyra (Walker) (Diptera :Tephritidae); Dimbi S et al.; The pathogenicity of two isolates of Beauveria bassiana and 12 of Metarhizium anisopliae towards adult fruit flies, Ceratitis capitata and Ceratitis rosa var . fasciventris was tested in the laboratory . Fruit flies were exposed to dry conidia evenly spread on velvet material covering the inner side of a cylindrical plastic tube . All isolates tested were pathogenic to both species of fruit flies . Mortality ranged from 7 to 100% in C . capitata and from 11.4 to 100% in C . rosa var . fasciventris at 4 days post-inoculation . Six isolates, M . anisopliae ICIPE 18, 20, 32, 40, 41 and 62, were highly pathogenic to both C . capitata and C . rosa var . fasciventris . The LT90 values of the most pathogenic isolates ranged between 3-4 days in both insects . Because of the difficulties in rearing C . cosyra, only the isolates that were highly pathogenic to both C . rosa var . fasciventris and C . capitata were tested against adult C . cosyra . They caused mortality of between 72-78% at 4 days post-inoculation . The LT90 values in all the isolates did not exceed 4 days . One of the most pathogenic isolates, M . anisopliae ICIPE 20, was evaluated against C . capitata and C . rosa var . fasciventris in cage experiments using three autoinoculators (maize cob, cheesecloth and Petri dish) in an autoinoculative device consisting of plastic mineral bottle . Mortality of between 70-93% was observed in flies of both species that were captured from the cages and held under laboratory conditions . These results indicate the possibility of fruit fly suppression with entomopathogenic fungi using an autoinoculative device. J Oral Maxillofac Surg, 2003 Dec, 61(12), 1455 - 62 Bone tissue engineering by primary osteoblast-like cells in a monolayer system and 3-dimensional collagen gel; Wiesmann HP et al.; PURPOSE: To engineer living bone tissue in vitro, bone cells must be multiplied and differentiated in cell culture . Osteoblasts are known to be the crucial cells responsible for the bone modeling process . Periosteal-derived osteoblasts were therefore cultured for up to 3 weeks in Petri dishes as well as in a 3-dimensional collagen gel . METHODS: Proliferation, migration, and differentiation of cells as well as the synthesis of extracellular matrix proteins were monitored during the culture period by histology, electron microscopy, and immunohistochemistry . Mineral formation was investigated by electron diffraction studies and element analysis . RESULTS: Osteoblasts proliferated and migrated in Petri dishes as well as in the collagen gel without loss of viability during the whole experimental period . They demonstrated a mature osteoblast phenotype as indicated by the synthesis of a bone-like extracellular matrix . They formed an extracellular matrix containing osteocalcin, osteonectin, and newly synthesized collagen type I in both environments . Mineral formation was seen in colocalization with the bone-like extracellular matrix proteins in Petri dishes . Microanalytical investigations revealed a matrix vesicle-mediated mineral formation at early stages of culture . CONCLUSIONS: Our cell culture confirmed the ability to multiplicate differentiated and viable osteoblast-like cells in 2- and 3-dimensional space . Additionally, bone-like mineralization can be induced by primary osteoblasts in monolayer culture . The data suggest that this approach can be used as a tool in bone tissue engineering. Appl Environ Microbiol, 2003 Dec, 69(12), 7210 - 5 Laboratory cultivation of widespread and previously uncultured soil bacteria; Joseph SJ et al.; Most soil bacteria belong to family-level phylogenetic groups with few or no known cultivated representatives . We cultured a collection of 350 isolates from soil by using simple solid media in petri dishes . These isolates were assigned to 60 family-level groupings in nine bacterial phyla on the basis of a comparative analysis of their 16S rRNA genes . Ninety-three (27%) of the isolates belonged to 20 as-yet-unnamed family-level groupings, many from poorly studied bacterial classes and phyla . They included members of subdivisions 1, 2, 3, and 4 of the phylum Acidobacteria, subdivision 3 of the phylum Verrucomicrobia, subdivision 1 of the phylum Gemmatimonadetes, and subclasses Acidimicrobidae and Rubrobacteridae of the phylum ACTINOBACTERIA: In addition, members of 10 new family-level groupings of subclass Actinobacteridae of the phylum Actinobacteria and classes Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria of the phylum Proteobacteria were obtained . The high degree of phylogenetic novelty and the number of isolates affiliated with so-called unculturable groups show that simple cultivation methods can still be developed further to obtain laboratory cultures of many phylogenetically novel soil bacteria. Cell Tissue Res, 2004 Feb, 315(2), 271 - 7 Epub 2003 Nov 26. Implantation in vitro: co-culture of rat blastocyst and epithelial cell vesicles; Tiwari R et al.; Implantation of blastocysts involves conversion of maternal and embryonic cell surfaces from a nonadhesive to an adhesive state in response to the internally driven developmental program or to externally generated factors . However, the intricacies of the cellular and subcellular changes that promote the attachment are not known, because these changes are difficult to determine in situ because of the nonaccessibility of the site . To overcome this, an in vitro model of implantation was developed by co-culturing rat blastocysts and uterine epithelial cells of the same gestational age (day 5 postcoitum; plug day as day 1) in drops hanging from the lid of a Petri dish . The system was used to study the changes on the surface membranes of the cells of the trophectoderm and uterine epithelium and to evaluate the antiadhesive activity of the newly designed test substances . The isolated epithelial cell vesicles were co-cultured with zona-free blastocysts in the microdrops (40-50 microl) hanging from the lid of a 60-mm Petri dish . The lid was placed over the lower dish, which was presaturated with the medium . The culture was examined 48 h later to determine the site of adhesion of epithelial cell vesicles with the trophoblasts lining the blastocyst . The cell-cell adhesion was monitored on a computerized image analyzer . To validate the adhesion of blastocysts and epithelial cell vesicles in co-culture, the expression of a cell adhesion molecule, uvomorulin, was stu