Occurrence of 16s rRNA methylase ArmA producing Enterobacteriaceae in a general hospital in Warsaw, Poland

Resistance to gentamicin, amikacin and kanamycin was screened in 270 clinical isolates of Enterobacteriaceae originated from April 19 to May 19, 2010 in a regular hospital in Warsaw, Poland. Most of the isolated bacteria were considered pathogenic. Nineteen isolates (7%) were simultaneously resistant to two or three of the tested aminoglycosides. MICs of the three aminoglycosides ranged form 128 to 1024 mcg/ml for six isolates. These isolates were suspected to produce 16S rRNA methylase. Genes encoding for three methylases reported in Europe: ArmA, RmtB and RmtC were searched by PCR. The armA gene was detected in all of the six isolates. This group encompassed Enterobacter cloacae (n=4), Klebsiella pneumoniae (n=1) and Proteus mirabilis (n=1). Five isolates of this group carried the bla(CAX-M) gene for CTX-M type ESBL. The remaining isolate E. cloacae DM0340 was ESBL negative and lacked bla(CRX-M) that may suggest an altered genetic environment of the armA gene in this isolate. Our results showed that 2.2% of the tested isolates produced 16S rRNA methylase ArmA. This finding may argue for a high incidence of ArmA producing Enterobacteriaceae in Poland when compared to reports from other European countries.     

Morphological and physiological responses of canola (Brassica napus) siliquas and seeds to UVB and CO2 under controlled environment conditions

Combined effects of UVB radiation and CO₂ concentration on plant reproductive parts have received little attention. We studied morphological and physiological responses of siliquas and seeds of canola (Brassica napus L. cv. 46A65) to UVB and CO₂ under four controlled experimental conditions: UVB radiation (4.2 kJ m⁻² d⁻¹) with ambient level of CO₂ (370 μmol mol⁻¹) (control); UVB radiation (4.2 kJ m⁻² d⁻¹) with elevated level of CO₂ (740 μmol mol⁻¹); no UVB radiation (0 kJ m⁻² d⁻¹) with ambient level of CO₂ (370 μmol mol⁻¹); and no UVB radiation (0 kJ m⁻² d⁻¹) with elevated level of CO₂ (740 μmol mol⁻¹). UVB radiation affected the outer appearance of siliquas, such as colour, as well as their anatomical structures. At both CO₂ levels, the UVB radiation of 4.2 kJ m⁻² d⁻¹ reduced the size of seeds, which had different surface patterns than those from no UVB radiation. At both CO₂ levels, 4.2 kJ m⁻² d⁻¹ of UVB decreased net CO₂ assimilation (A_N) and water use efficiency (WUE), but had no effect on transpiration (E). Elevated CO₂ increased A_N and WUE, but decreased E, under both UVB conditions. At both CO₂ levels, the UVB radiation of 4.2 kJ m⁻² d⁻¹ decreased chlorophyll fluorescence, total chlorophyll (Chl), Chl a and Chl b, but had no effect on the ratio of Chl a/b and the concentration of UV-screening pigments. Elevated CO₂ increased total Chl and the concentration of UV-screening pigments under 4.2 kJ m⁻² d⁻¹ of UVB radiation. Neither UVB nor CO₂ affected wax content of siliqua surface. Many significant relationships were found between the above-mentioned parameters. This study revealed that UVB radiation exerts an adverse effect on canola siliquas and seeds, and some of the detrimental effects of UVB on these reproductive parts can partially be mitigated by CO₂.     

Anterior regions of monkey parietal cortex process visual 3D shape

The intraparietal cortex is involved in the control of visually guided actions, like reach-to-grasp movements, which require extracting the 3D shape and position of objects from 2D retinal images. Using fMRI in behaving monkeys, we investigated the role of the intraparietal cortex in processing stereoscopic information for recovering the depth structure and the position in depth of objects. We found that while several areas (CIP, LIP, and AIP on the lateral bank; PIP and MIP on the medial bank) are activated by stereoscopic stimuli, AIP and an adjoining portion of LIP are sensitive only to depth structure. Furthermore, only these two regions are sensitive to both the depth structure and the 2D shape of small objects. These results indicate that extracting 3D spatial information from stereo involves several intraparietal areas, among which AIP and anterior LIP are more specifically engaged in extracting the 3D shape of objects.     

Nf1 and Gmcsf interact in myeloid leukemogenesis

The NF1 tumor suppressor gene encodes neurofibromin, a GTPase-activating protein (GAP) for p21ras (Ras). Children with NF1 are predisposed to juvenile myelomonocytic leukemia (JMML). Some heterozygous Nf1 mutant mice develop a similar myeloproliferative disorder (MPD), and adoptive transfer of Nf1-deficient fetal liver cells consistently induces this MPD. Human JMML and murine Nf1-deficient cells are hypersensitive to granulocyte-macrophage colony-stimulating factor (GM-CSF) in methylcellulose cultures. We generated hematopoietic cells deficient in both Nf1 and Gmcsf to test whether GM-CSF is required to drive excessive proliferation of Nf1-/- cells in vivo. Here we show that GM-CSF play a central role in establishing and maintaining the MPD and that recipients engrafted with Nf1-/- Gmcsf-/- hematopoietic cells are hypersensitive to exogenous GM-CSF.     

A rapid effect of applied brassinolide on abscisic acid concentrations in <Emphasis Type="Italic">Brassica napus</Emphasis> leaf tissue subjected to short-term heat stress

Three-week old canola (Brassica napus L.) seedlings grown at 20/16°C (day/night) were subjected to short-term (4 and 8 h) heat stress (45°C) or maintained at a normal temperature of 20°C. Half of the plants under each treatment received a 10⁻⁶ M solution of brassinolide (BL) 1 h prior to beginning the temperature treatments. The concentration (ng/g dry weight) of endogenous abscisic acid (ABA) was subsequently determined in young leaves via the stable isotope dilution method. Applied BL had no effect on endogenous ABA for plants maintained at normal temperatures. However, ABA concentration was significantly elevated by heat stress alone and doubled by heat stress + BL. These results suggest that the well-known enhancement of tolerance to high temperature stress that can be obtained by BL or 24-epi-BL applications may be caused by a brassinosteroid-induced elevation in endogenous ABA concentration.     

Identification of Risks in the Life Cycle of Nanotechnology‐Based Products

In order to realize the projected market potential of nanotechnology, the environmental, health, and safety (EHS) uncertainties posed by a nano‐product (i.e., a nanotechnology‐enabled product) need to be characterized through the identification of risks and opportunities in early stages of product development. We present a methodology to identify risks from nano‐products using a scenario analysis approach that allows for expert elicitation on a set of preidentified use and disposal scenarios and what we have labeled “risk triggers” to obtain scores on their likelihood of occurrence and severity. Use and disposal scenarios describe product life‐cycle stages that could result in risk attributed to the nano‐product, whereas risk triggers are particular to nanoparticle properties. These are potential risks, as the risk assessment community is currently debating the specific risks attributed to nanotechnology. Through such a framework, our goal is to identify which products pose greater risks, where these risks occur in the product life cycle, and the impacts of these environmental risks on society. The comparison of risk triggers across nano‐products allows relative risk ranking on axes of exposure‐ and hazard‐related risk triggers. For the specific case of air fresheners, areas of acute risks resulted from bioavailability of nanoparticles in air release and water entrainment exposure scenarios; catalytic activity of nanoparticles in inhalation and air release exposure scenarios; the harmful effects due to the antibacterial property on useful bacteria particularly in susceptible populations; and, finally, risks from the lack of nanoparticle coating stability in air release scenarios.     

Methane emissions from six crop species exposed to three components of global climate change: temperature, ultraviolet-B radiation and water stress

The l ‐ascorbate (AsA) content and the expression of six l ‐galactose pathway‐related genes were analyzed in peach flesh during fruit development. Fluctuation of AsA during peach fruit development was divided into four phases based on the overall total AsA (T‐AsA) content per fruit: AsA I, 0–36 days after full bloom (DAFB); AsA II, 37–65 DAFB; AsA III, 66–92 DAFB and AsA IV, 93–112 DAFB. Phase AsA III was a lag phase for AsA accumulation, but did not coincide with the lag phase for fruit development. The T‐AsA concentration was highest at the early stage until 21 DAFB [2–3μmol per gram of fresh weight (g^?1 FW)], and decreased to 1/4 and 1/15 of this value at 50 and 92 DAFB, respectively. T‐AsA then remained at 0.15–0.20μmol g^?1 FW until harvest at 112 DAFB. More than 90% of the T‐AsA was in the reduced form until 21 DAFB. The proportion of reduced form of AsA then decreased concomitantly with the decrease in AsA concentration. To determine the main pathway of AsA biosynthesis and the AsA biosynthetic capacity of peach flesh, several precursors were incubated with immature whole fruit (59 DAFB). The AsA concentration increased markedly with l ‐galactono‐1,4‐lactone or l ‐galactose (Gal), but d ‐galacturonate and l ‐gulono‐1,4‐lactone failed to increase AsA, indicating dominance of the Gal pathway and potent AsA biosynthetic capabilities in immature peach flesh. The expression of genes involved in the last six steps of the Gal pathway was measured during fruit development. The genes studied included GDP‐d ‐mannose pyrophosphorylase (GMPH), GDP‐ d ‐mannose‐3′,5′‐epimerase (GME), GDP‐ l ‐galactose guanylyltransferase (GGGT), l ‐galactose‐1‐phosphate phosphatase (GPP), l ‐galactose‐1‐dehydrogenase (GDH) and l ‐galactono‐1,4‐lactone dehydrogenase (GLDH). GMPH, GME and GGGT had similar expression patterns that peaked at 43 DAFB. GPP, GDH and GLDH also had similar expression patterns that peaked twice at 21 and 91 DAFB, although the expression of GDH was quite low. High level of T‐AsA concentration was roughly correlated with the level of gene expression in the early period of fruit development (AsA I), whereas no such relationships were apparent in the other periods (e.g. AsA III and IV). On the basis of these findings, we discuss the regulation of AsA biosynthesis in peach fruit.     

PCR-restriction fragment length polymorphism assay (PCR-RFLP) as an useful tool for detection of mutation in gyrA gene at 86-THR position associated with fluoroquinolone resistance in Campylobacter jejuni

The aim of this study was to investigate the mutations in gyrA gene at Thr-86 position in fluoroquinolone resistant C. jejuni clinical isolates (2003-2005). The change of Thr to Ile at 86 position is associated with high-level resistance to fluoroquinolone in C. jejuni. Thirty five (58%) of 65 C. jejuni strains were found to be resistant to ciprofloxacin using E-test method. PCR-RFLP technique with the RsaI enzyme was used for the identification of mutation in gyrA gene. The primers spanning a part of the fluoroquinolone resistance determining region (QRDR) were designed based on the article of Alonso et al. and the gyrA sequence of C. jejuni (Gen Bank accession number LO4566). One of this primer had mismatch introduced at the second nucleotide from 3' end of the primer what gives an artificial RsaI cleavage site. All of the ciprofloxacin-resistant isolates contained a single)point mutation in the gyrA gene: the replacement of Thr 86 by Ile. The results showed that PCR-RFLP is a rapid and simple method for the detection of the high-level fluoroquinolone resistance in C. jejuni.     

Mutation to Bax beyond the BH3 domain disrupts interactions with pro-survival proteins and promotes apoptosis

Pro-survival members of the Bcl-2 family of proteins restrain the pro-apoptotic activity of Bax, either directly through interactions with Bax or indirectly by sequestration of activator BH3-only proteins, or both. Mutations in Bax that promote apoptosis can provide insight into how Bax is regulated. Here, we describe crystal structures of the pro-survival proteins Mcl-1 and Bcl-x(L) in complex with a 34-mer peptide from Bax that encompasses its BH3 domain. These structures reveal canonical interactions between four signature hydrophobic amino acids from the BaxBH3 domain and the BH3-binding groove of the pro-survival proteins. In both structures, Met-74 from the Bax peptide engages with the BH3-binding groove in a fifth hydrophobic interaction. Various Bax Met-74 mutants disrupt interactions between Bax and all pro-survival proteins, but these Bax mutants retain pro-apoptotic activity. Bax/Bak-deficient mouse embryonic fibroblast cells reconstituted with several Bax Met-74 mutants are more sensitive to the BH3 mimetic compound ABT-737 as compared with cells expressing wild-type Bax. Furthermore, the cells expressing Bax Met-74 mutants are less viable in colony assays even in the absence of an external apoptotic stimulus. These results support a model in which direct restraint of Bax by pro-survival Bcl-2 proteins is a barrier to apoptosis.     

The occurrence and characterisation of oxy-imino-beta-lactams resistant strains among Salmonella enterica subsp. enterica isolated in Poland

Extended-spectrum beta-lactamases (ESBLs) are enzymes manifesting considerable hydrolyzing activity on a wide variety of beta-lactam antibiotics including oxyiminocephalosporins and aztreonam. In the study reported here we investigated the types of ESBL produced by Salmonella enterica subsp. enterica strains isolated from clinical samples in the microbiological laboratories of sanitary-epidemiological units in Poland from 1999 to 2004. Among 239 ampicillin-resistant Salmonella enterica subsp. enterica strains isolated from clinical samples in the microbiological laboratories of sanitary-epidemiological units in Poland, 68 isolates of oximino-beta-lactams resistant of 6 serovars were found. There were 16 epidemiological unrelated strains (6 isolates of S. Enteritidis, 5 isolates of S. Thompson, 3 isolates of S. Typhimurium, one-fold isolate of S. Muenster and S. enterica 1,9,12:-:-) coming from different areas of country and 52 epidemiologically related isolates of S. Oranienburg, coming from a prolonged outbreak in an orphanage in Lód?. All the strains were identified as the ESBLs producers. The molecular analysis revealed that most of them expressed CTX-M-3 ESBL which is widely observed in Poland and additional enzyme TEM-1. All tested isolates of S. Thompson and one of three S. Typhimurium isolates were found to produce SHV-5 ESBL. This is the first report regarding the presence of SHV-5 in the genus Salmonella in Poland.