Clusters of ligands on dendrimer surfaces

The development of methodology that is designed to allow a significant increase in the patterning and in the functionalization of the dendrimer is the ultimate goal of the research described here. Glycoside clusters based on TRIS were formed using click chemistry and were attached to PAMAM dendrimers. A series of dendrimers bearing tris-mannoside and an ethoxyethanol group was synthesized, and the binding interactions of these dendrimers with Concanavalin A were evaluated using inhibition ELISAs. The results of the inhibition ELISAs suggest that tris-mannoside clusters can replace individual sugars on the dendrimer without loss of function. Since tris-mannoside clustering allows for a redistribution of the dendrimers’ surface functionalities, from this chemistry one can envision patterned dendrimers that incorporate multiple groups to increase the function and utility of the dendrimer.     

Alternative therapies in Staphylococcus aureus diseases

Staphylococcus aureus is a common pathogen responsible for health-care-associated infections as well as community acquired ones. It is the etiological factor of a wide spectrum of infections. Therapeutic problems are caused by resistance of S. aureus to many antibiotics, specifically to methicillin (methicillin-resistant S. aureus, MRSA). In such cases a limited spectrum of antibiotics may be used and prolonged hospitalization is costly. Hence, there is an urgent need for the development of alternative antibiotic therapeutics. This work reviews the current knowledge concerning prospective treatment of staphylococcal diseases.     

Individual Somatic H1 Subtypes Are Dispensable for Mouse Development Even in Mice Lacking the H10 Replacement Subtype

H1 linker histones are involved in facilitating the folding of chromatin into a 30-nm fiber. Mice contain eight H1 subtypes that differ in amino acid sequence and expression during development. Previous work showed that mice lacking H1(0), the most divergent subtype, develop normally. Examination of chromatin in H1(0-/-) mice showed that other H1s, especially H1c, H1d, and H1e, compensate for the loss of H1(0) to maintain a normal H1-to-nucleosome stoichiometry, even in tissues that normally contain abundant amounts of H1(0) (A. M. Sirotkin et al., Proc. Natl. Acad. Sci. USA 92:6434-6438, 1995). To further investigate the in vivo role of individual mammalian H1s in development, we generated mice lacking H1c, H1d, or H1e by homologous recombination in mouse embryonic stem cells. Mice lacking any one of these H1 subtypes grew and reproduced normally and did not exhibit any obvious phenotype. To determine whether one of these H1s, in particular, was responsible for the compensation present in H1(0-/-) mice, each of the three H1 knockout mouse lines was bred with H1(0) knockout mice to generate H1c/H1(0), H1d/H1(0), or H1e/H1(0) double-knockout mice. Each of these doubly H1-deficient mice also was fertile and exhibited no anatomic or histological abnormalities. Chromatin from the three double-knockout strains showed no significant change in the ratio of total H1 to nucleosomes. These results suggest that any individual H1 subtype is dispensable for mouse development and that loss of even two subtypes is tolerated if a normal H1-to-nucleosome stoichiometry is maintained. Multiple compound H1 knockouts will probably be needed to disrupt the compensation within this multigene family.     

Possible Involvement of GABAA and GABAB Receptors in the Inhibitory Action of Lindane on Transmitter Release from Cerebellar Granule Neurons

Cerebellar granule cells in culture express receptors for GABA belonging to the GABA_A and GABA_B classes. In order to characterize the ability of the insecticide lindane to interact with these receptors cells were grown in either plain culture media or media containing 150 M THIP as this is known to influence the properties of both GABA_A and GABA_B receptors. It was found that lindane regardless of the culture condition inhibited evoked (40 mM K⁺) release of neurotransmitter ([³H]D-aspartate as label for glutamate). In naive cells both GABA_A and GABA_B receptor active drugs prevented the inhibitory action of lindane but in THIP treated cultures none of the GABA_A and GABA_B receptor active drugs had any effect on the inhibitory action of lindane. This lack of effect was not due to inability of baclofen itself to inhibit transmitter release. It is concluded that lindane dependent on the state of the GABA_A and GABA_B receptors is able to indirectly interfere with both GABA_A and GABA_B receptors. In case of the latter receptors it was shown using [³H]baclofen to label the receptors that lindane could not displace the ligand confirming that lindane is likely to exert its action at a site different from the agonist binding site.     

Impact of picolinic acid on the chromium accumulation in fodder radish and komatsuna

Effects of picolinic acid (2-pyridinecarboxylic acid) and chromium(III) picolinate was studied on the chromium (Cr) accumulation of fodder radish (Raphanus sativus L. convar. oleiformis Pers., cv. Leveles olajretek) and komatsuna (Brassica campestris L. subsp. napus f. et Thoms. var. komatsuna Makino, cv. Kuromaru ) grown in a pot experiment. Control cultures, grown in an uncontaminated soil (UCS; humous sand with pH_KCl 7.48, sand texture with 12.4% clay+silt content, organic carbon 0.56%, CaCO₃ 2.2%, CEC 6.2 cmol_c kg^–1, Cr 10.6 mg kg^–1), accumulated low amounts of chromium (less than 5.4 g g^–1) in their roots or shoots. When this UCS was artificially contaminated with 100 mg kg^–1 Cr (CrCl₃) later picolinic acid treatment promoted the translocation of chromium into the shoots of both species. In fodder radish shoots Cr concentration reached 30.4 g g^–1 and in komatsuna shoots 44.5 g g^–1. Application of ethylene diamine tetra-acetic acid (EDTA) to this Cr contaminated soil had similar effect to picolinic acid. When the UCS was amended with leather factory sewage sediment (which resulted in 853 mg kg^–1 Cr in soil), Cr mobilization was observed only after repeated soil picolinic acid applications. From a galvanic mud contaminated soil (brown forest soil with pH_KCl 6.77, loamy sand texture with 26.6% clay+silt content, organic carbon 1.23%, CaCO₃ 0.7%, CEC 24.5 cmol_c kg^–1, Cd 5.0 mg kg^–1, Cr 135 mg kg^–1, and Zn 360 mg kg^–1) the rate of Cr mobilization was negligible, only a slight increase was observed in Cr concentration of fodder radish shoots after repeated picolinic acid treatments of soil. Presumably picolinic acid forms a water soluble complex (chromium(III) picolinate) with Cr in the soil, which promotes translocation of this element (and also Cu) into the shoots of plants. The rate of complex formation may be related to the binding forms and/or concentration of Cr in soil and also to soil characteristics (i.e. pH, CEC), since the rate of Cr translocation was the following: artificially contaminated soil > leather factory sewage sediment amended soil > galvanic mud contaminated soil. Four times repeated 10 mg kg^–1 chromium(III) picolinate application to UCS multiplied the transport of chromium to shoots, as compared to single 10 mg kg^–1 CrCl₃ treatment. This also suggests that chromium(III) picolinate is forming in the picolinic acid treated Cr-contaminated soils, and plants more readily accumulates and translocates organically bound Cr than ionic Cr. Picolinic acid promotes Cr translocation in soil-plant system. This could be useful in phytoextraction (phytoremediation) of Cr contaminated soils or in the production of Cr enriched foodstuffs.     

Neurotoxicity of Lindane and Picrotoxin: Neurochemical and Electrophysiological Correlates in the Rat Hippocampus In Vivo

In the present study, we compared in vivo changes of extracellular amino acid levels and nucleotide derivatives to a single ip dose of lindane (10-60 mg/kg) and picrotoxin (5 mg/kg) in the hippocampus of halothane anaesthetized rat by microdialysis-coupled HPLC analysis. Brain activity was monitored by EEG. The effects of lindane and picrotoxin on EEG pattern of rats as well as on hippocampal amino acid and nucleotide status were studied in 0-50 min, 50-100 min and 100-150 min periods post-dosing. Significant decreases in Glu and Asp were found after picrotoxin treatment. After 50-100 min post-dosing, hippocampal hypoxanthine and inosine levels increased to both lindane (10 mg/kg) and picrotoxin whereas xanthine and uridine levels increased to picrotoxin, only. Lindane elicited a dose-dependent occurrence of negative spikes accompanied with rhythmic activity at 4-5 Hz. The picrotoxin-induced 4-5 Hz activity did not display negative sharp waves and was accompanied by 10 Hz oscillations.     

Acoustic accelerometry reveals diel activity patterns in premigratory Port Jackson sharks

Distinguishing the factors that influence activity within a species advances understanding of their behavior and ecology. Continuous observation in the marine environment is not feasible but biotelemetry devices provide an opportunity for detailed analysis of movements and activity patterns. This study investigated the detail that calibration of accelerometers measuring root mean square (RMS) acceleration with video footage can add to understanding the activity patterns of male and female Port Jackson sharks (Heterodontus portusjacksoni) in a captive environment. Linear regression was used to relate RMS acceleration output to time‐matched behavior captured on video to quantify diel activity patterns. To validate captive data, diel patterns from captive sharks were compared with diel movement data from free‐ranging sharks using passive acoustic tracking. The RMS acceleration data showed captive sharks exhibited nocturnal diel patterns peaking during the late evening before midnight and decreasing before sunrise. Correlation analysis revealed that captive animals displayed similar activity patterns to free‐ranging sharks. The timing of wild shark departures for migration in the late breeding season corresponded with elevated diel activity at night within the captive individuals, suggesting a form of migratory restlessness in captivity. By directly relating RMS acceleration output to activity level, we show that sex, time of day, and sex‐specific seasonal behavior all influenced activity levels. This study contributes to a growing body of evidence that RMS acceleration data are a promising method to determine activity patterns of cryptic marine animals and can provide more detailed information when validated in captivity.     

Receptor Tyrosine Kinase EphA5 Is a Functional Molecular Target in Human Lung Cancer

Lung cancer is often refractory to radiotherapy, but molecular mechanisms of tumor resistance remain poorly defined. Here we show that the receptor tyrosine kinase EphA5 is specifically overexpressed in lung cancer and is involved in regulating cellular responses to genotoxic insult. In the absence of EphA5, lung cancer cells displayed a defective G₁/S cell cycle checkpoint, were unable to resolve DNA damage, and became radiosensitive. Upon irradiation, EphA5 was transported into the nucleus where it interacted with activated ATM (ataxia-telangiectasia mutated) at sites of DNA repair. Finally, we demonstrate that a new monoclonal antibody against human EphA5 sensitized lung cancer cells and human lung cancer xenografts to radiotherapy and significantly prolonged survival, thus suggesting the likelihood of translational applications.     

Evaluation of biofilm production and prevalence of the icaD gene in methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains isolated from patients with nosocomial infections and carriers

Staphylococcus aureus is one of the most important etiological agents responsible for healthcare-associated infections and is capable of producing many virulence factors including biofilm. The aim of the present study was to analyze the correlation between the presence of the icaD and icaA genes and the ability to produce biofilm in vitro in 302 methicillin-resistant (MRSA) and 268 methicillin-sensitive S. aureus (MSSA) strains isolated in the Provincial Hospital in Gdansk. Presence of the icaD and icaA genes was detected by PCR and the ability to produce biofilm in vitro was measured both spectrophotometrically and via Congo Red Agar plate culture methods. We found that 91% of MRSA strains harbored the icaD gene. Moreover, all icaD-negative strains were icaA-positive. Of MRSA and MSSA strains, 47% and 69%, respectively, produced biofilm in vitro. The level of consistency between the two applied phenotypic methods was 96%. Additionally, we found that strains with the same biofilm status may be present in asymptomatic carriers and cause infections.