1,137 research outputs found

    Transformation in Micrococcus radiodurans and the ultraviolet sensitivity of its transforming DNA.

    Full text link

    Impact of Radiation Biology on Fundamental in Biology

    Full text link
    Research supported by OHER and its predecessors has as one of its major goals an understanding of the effects of radiation at low doses and dose rates on biological systems, so as to predict their effects on humans. It is not possible to measure such effects directly. They must be predicted from basic knowledge on how radiation affects cellular components such as DNA and membranes and how cells react to such changes. What is the probability of radiation producing human mutations and what are the probabilities of radiation producing cancer? The end results of such studies are radiation exposure standards for workers and for the general population. An extension of these goals is setting standards for exposure to chemicals involved in various energy technologies. This latter problem is much more difficult because chemical dosimetry is is a primitive state compared to radiation dosimetry

    Elastic and inelastic light scattering from single bacterial spores in an optical trap allows the monitoring of spore germination dynamics

    Get PDF
    Raman scattering spectroscopy and elastic light scattering intensity (ESLI) were used to simultaneously measure levels of Ca-dipicolinic acid (CaDPA) and changes in spore morphology and refractive index during germination of individual B. subtilis spores with and without the two redundant enzymes (CLEs), CwlJ and SleB, that degrade sporesâ peptidoglycan cortex. Conclusions from these measurements include: 1) CaDPA release from individual wild-type germinating spores was biphasic; in a first heterogeneous slow phase, Tlag, CaDPA levels decreased â ¼15% and in the second phase ending at Trelease, remaining CaDPA was released rapidly; 2) in L-alanine germination of wild-type spores and spores lacking SleB: a) the ESLI rose â ¼2-fold shortly before Tlag at T1; b) following Tlag, the ESLI again rose â ¼2-fold at T2 when CaDPA levels had decreased â ¼50%; and c) the ESLI reached its maximum value at â ¼Trelease and then decreased; 3) in CaDPA germination of wild-type spores: a) Tlag increased and the first increase in ESLI occurred well before Tlag, consistent with different pathways for CaDPA and L-alanine germination; b) at Trelease the ESLI again reached its maximum value; 4) in L-alanine germination of spores lacking both CLEs and unable to degrade their cortex, the time Î Trelease (Treleaseâ Tlag) for excretion of â ¥75% of CaDPA was â ¼15-fold higher than that for wild-type or sleB spores; and 5) spores lacking only CwlJ exhibited a similar, but not identical ESLI pattern during L-alanine germination to that seen with cwlJ sleB spores, and the high value for Î Trelease. Originally published Analytical Chemistry, Vol. 81, No. 10, May 200

    Constructing Fluorogenic Bacillus Spores (F-Spores) via Hydrophobic Decoration of Coat Proteins

    Get PDF
    Background: Bacterial spores are protected by a coat consisting of about 60 different proteins assembled as a biochemically complex structure with intriguing morphological and mechanical properties. Historically, the coat has been considered a static structure providing rigidity and mainly acting as a sieve to exclude exogenous large toxic molecules, such as lytic enzymes. Over recent years, however, new information about the coat’s architecture and function have emerged from experiments using innovative tools such as automated scanning microscopy, and high resolution atomic force microscopy. Principal Findings: Using thin-section electron microscopy, we found that the coat of Bacillus spores has topologically specific proteins forming a layer that is identifiable because it spontaneously becomes decorated with hydrophobic fluorogenic probes from the milieu. Moreover, spores with decorated coat proteins (termed F-spores) have the unexpected attribute of responding to external germination signals by generating intense fluorescence. Fluorescence data from diverse experimental designs, including F-spores constructed from five different Bacilli species, indicated that the fluorogenic ability of F-spores is under control of a putative germination-dependent mechanism. Conclusions: This work uncovers a novel attribute of spore-coat proteins that we exploited to decorate a specific layer imparting germination-dependent fluorogenicity to F-spores. We expect that F-spores will provide a model system to gai

    A model for transition of 5 '-nuclease domain of DNA polymerase I from inert to active modes

    Get PDF
    Bacteria contain DNA polymerase I (PolI), a single polypeptide chain consisting of similar to 930 residues, possessing DNA-dependent DNA polymerase, 3'-5' proofreading and 5'-3' exonuclease (also known as flap endonuclease) activities. PolI is particularly important in the processing of Okazaki fragments generated during lagging strand replication and must ultimately produce a double-stranded substrate with a nick suitable for DNA ligase to seal. PolI's activities must be highly coordinated both temporally and spatially otherwise uncontrolled 5'-nuclease activity could attack a nick and produce extended gaps leading to potentially lethal double-strand breaks. To investigate the mechanism of how PolI efficiently produces these nicks, we present theoretical studies on the dynamics of two possible scenarios or models. In one the flap DNA substrate can transit from the polymerase active site to the 5'-nuclease active site, with the relative position of the two active sites being kept fixed; while the other is that the 5'-nuclease domain can transit from the inactive mode, with the 5'-nuclease active site distant from the cleavage site on the DNA substrate, to the active mode, where the active site and substrate cleavage site are juxtaposed. The theoretical results based on the former scenario are inconsistent with the available experimental data that indicated that the majority of 5'-nucleolytic processing events are carried out by the same PolI molecule that has just extended the upstream primer terminus. By contrast, the theoretical results on the latter model, which is constructed based on available structural studies, are consistent with the experimental data. We thus conclude that the latter model rather than the former one is reasonable to describe the cooperation of the PolI's polymerase and 5'-3' exonuclease activities. Moreover, predicted results for the latter model are presented

    Procjena izloženosti UV zračenju tijekom ljetnih mjeseci u Hrvatskoj s pomoću jednostavne približne formule

    Get PDF
    The Tropospheric Ultraviolet-Visible (TUV) model, version 4.2 developed by Madronich (2003) was usedto estimate the extent of ultraviolet (UV) exposure of general population in Croatia over the summer. Solarnoon values (13 h local time, CEST) of the ultraviolet index (UVI) for the period April to October 2004 were calculated for 61 cities in Croatia. The results showed that the risk of sunburn at 13 h local time inclear weather was high between April and September (UVI >7) and very high in July (UVI >10). In July, the UVI exceeded 8 between 11 h and 15 h local time. In this study, we developed a simple approximate formula to estimate UVI. The formula includes data on the time, date, altitude and clouds. The difference between our estimate and the TUV model for the summer months of June, July and August at 10 h to16 h local time was less than 10 %.Tropospheric Ultraviolet-Visible (TUV) model, verzija 4.2 autora S. Madronicha (2003.) upotrijebljen je zaprocjenu izloženosti ultraljubičastom (UV) zračenju stanovništva u Hrvatskoj. Podnevne vrijednosti (13 hprema lokalnom vremenu) ultraljubičastog indeksa (UVI) izračunane su za 61 mjesto u Hrvatskoj za razdobljetravanj - listopad. Rezultati pokazuju da je u 13 h prema lokalnom vremenu rizik od nastanka opeklina izazvanih sunčevim zračenjem u danima bez naoblake visok između travnja i rujna (UVI > 7) te da je rizikvrlo visok tijekom srpnja (UVI >10). U srpnju tijekom dana UV indeks prelazi vrijednost 8 između 11 h i15 h prema lokalnom vremenu. U ovom radu za procjenu UV indeksa razvijena je jednostavna približna formula. Formula omogućava procjenu UV indeksa na temelju podataka o datumu, satu, nadmorskoj visini i naoblaci. Prilikom usporedbe rezultata dobivenih formulom i točnih rezultata dobivenih TUV modelom za ljetne mjesece lipanj, srpanj i kolovoz te razdoblje od 10 h do 16 h među rezultatima dobivena je razlikamanja od 10 %

    Analysis of Clostridium beijerinckii NCIMB 8052's transcriptional response to ferulic acid and its application to enhance the strain tolerance

    Get PDF
    Background: Plant-based cellulose presents the best source of renewable sugars for biofuel production. However, the lignin associated with plant cellulose presents a hurdle as hydrolysis of this component leads to the production of inhibitory compounds, such as ferulic acid. Results: The impacts of ferulic acid, a phenolic compound commonly found in lignin hydrolysates, on the growth, solvent production, and transcriptional responses of Clostridium beijerinckii NCIMB 8052 were determined. Addition of ferulic acid to growing cultures resulted in a decrease in the growth and solvent production by 30% and 25%, respectively, when compared to the control cultures. To better understand the toxicity of this compound, microarray analyses were performed using samples taken from these cultures at three different growth states. Several gene ontology terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were identified showing significant change at each status, including ATP-binding cassette (ABC) transporters, two component system, and oxidoreductase activity. Moreover, genes related with efflux systems and heat shock proteins were also strongly up-regulated. Among these, expression of the groESL operon was induced by more than fourfold and was consequently selected to improve C. beijerinckii tolerance to ferulic acid. Real-time quantitative PCR (RT-qPCR) analysis confirmed that C. beijerinckii harboring the plasmid, pSAAT-ptb_Gro, had a two-to fivefold increased groESL operon expression during growth of these cultures. Moreover, this strain was more tolerant to ferulic acid as the growth of this recombinant strain and its bioconversion of glucose into solvents were both improved. Conclusions: Using transcriptomics, we identified numerous genes that are differentially expressed when C. beijerinckii cultures were exposed to ferulic acid for varying amounts of time. The operon expressing groESL was consistently up-regulated, suggesting that this gene cluster may contribute to strain tolerance. This was confirmed as recombinant cultures showed both an enhanced growth and solvent yield in the presence of 0.5 g/L ferulic acidopen00
    corecore