Genetic determinants of resistance of hospital-associated strains of <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis> to β-lactam antibiotics isolated in neonates

According to the results of analysis of whole genome sequencing, the presence of genes having resistance to β-lactam antibiotics in hospital-associated strains of Klebsiella pneumoniae was studied. The strains were isolated from neonatal intensive care units. The data obtained were compared with the results of antimicrobial susceptibility testing of isolated microorganisms. Among other strains resistant to cephalosporins, the dominance of genes of CTX-M-type extended-spectrum β-lactamases was shown. It was revealed that one of eight strains phenotypically resistant and moderately resistant to carbapenems have the blaOXA-48 carbapenemase gene.     

Amplification of the phage lambda DNA sequence by polymerase chain reaction using thermostable DNA polymerase]

The efficiency of phage DNA amplification by the method of polymerase chain reaction (PCR) with Tth DNA-polymerase was studied for optimization of PCR conditions. The effect on amplification efficiency of medium ionic strength and pH, the presence of univalent cations, detergents, gelatin, ATP, pyrophosphate, SH-reagents and ratio of concentrations of Mg and dNTPs, primers and template was studied. It has been found that a pH optimum for PCR with Tth DNA-polymerase varies from 8.5 to 9.0. An ionic strength optimum for PCR is about 0.08. The influence of univalent cations on the activity of Tth DNA-polymerase can be expressed as NH4+ greater than Na+ greater than K+. 0.01% Tween-20 significantly increases the efficiency of PCR and 0.01% gelatin inhibits it. Addition of ATP, pyrophosphate, SH-reagents to the reaction mixture did not increase the yield of PCR product. It has been also shown that for the given PCR-system an optimum Mg/dNTPs molar ratio is within the range of 1.5-2.0. An optimum concentration of each of the pair of primers for this PCR-system is about 0.3 microM. The possibility of PCR-amplification of 500-8500 b.p. DNA fragments has been demonstrated.     

Night,day, sunrise,sunset: do fish under snow and ice recognize the difference?

1. Although boreal lakes are ice‐covered for several months annually, little is known about the behaviour of fish under ice. To consider the reasons for diel vertical migrations (DVM) it is important to compare periods under ice as opposed to under open water. Echosounding provides a tool for non‐intrusive continuous monitoring of fish, even in winter. 2. Changes in the vertical distribution of fish through six 48‐h periods were monitored using a stationary, mounted echosounder that beamed vertically either from the bottom up or from the surface down from February to April, 2003. The up‐beaming and down‐beaming transducers were run alternately for 24 h each over the 48‐h period. Standard echo analysis software was used to detect fish traces and estimate the vertical and temporal distribution of fish abundance. Fish were sampled with a winter seine. 3. Prominent diel vertical migration in response to changing light level was detected throughout the study period (late winter to spring). Fish were highest in the water column at sunset and sunrise. In daylight, most detected fish were well below 10‐m depth. The number of fish detected was greatest during the night when they occurred throughout almost the whole water column, sometimes with a considerable number very close to the ice. Fish were mostly vendace and whitefish. 4. It became evident from data from the up‐beaming transducer that at night fish may occupy the layer closest to the surface. These fish would not have been detected if we had only used the down‐beaming transducer. The overall pattern of DVM in winter was very similar to that in summer. The results support the suggestion that DVM is a genetically fixed behavioural trait responding to the contemporary level of illumination.     

Atmospheric impact of bioenergy based on perennial crop (reed canary grass,Phalaris arundinaceae,L.) cultivation on a drained boreal organic soil

Marginal organic soils, abundant in the boreal region, are being increasingly used for bioenergy crop cultivation. Using long‐term field experimental data on greenhouse gas (GHG) balance from a perennial bioenergy crop [reed canary grass (RCG), Phalaris arundinaceae L.] cultivated on a drained organic soil as an example, we show here for the first time that, with a proper cultivation and land‐use practice, environmentally sound bioenergy production is possible on these problematic soil types. We performed a life cycle assessment (LCA) for RCG on this organic soil. We found that, on an average, this system produces 40% less CO₂‐equivalents per MWh of energy in comparison with a conventional energy source such as coal. Climatic conditions regulating the RCG carbon exchange processes have a high impact on the benefits from this bioenergy production system. Under appropriate hydrological conditions, this system can even be carbon‐negative. An LCA sensitivity analysis revealed that net ecosystem CO₂ exchange and crop yield are the major LCA components, while non‐CO₂ GHG emissions and costs associated with crop production are the minor ones. Net bioenergy GHG emissions resulting from restricted net CO₂ uptake and low crop yields, due to climatic and moisture stress during dry years, were comparable with coal emissions. However, net bioenergy emissions during wet years with high net uptake and crop yield were only a third of the coal emissions. As long‐term experimental data on GHG balance of bioenergy production are scarce, scientific data stemming from field experiments are needed in shaping renewable energy source policies.     

Imipenem and cefepime in empirical therapy of infectious complications after radical surgeries in patients with invasive cancer of the urinary bladder]

Urinary tract injection (UTI) is one of the main causes of severe complications and death in oncologic patients and particularly in patients with invasive cancer of the urinary bladder. In our Clinic an up-to-date system for infection control recording all cases of UTI in oncologic patients is being exploited since 2001. The paper presents the data on our observations during the period from 2001 to 2003 concerning typical UTI in patients with invasive cancer of the urinary bladder, etiological agents of UTI and their susceptibility to a wide range of antibacterials, including imipenem and cefepime, a 4th generation cephalosporin, used for adequate therapy of UTI in oncourologic patients.     

Up-regulation of multiple serine proteinases during earthworm tail regeneration

Summary

Previous studies have shown that spatiotemporal regulation of extracellular matrix (ECM) by proteinases is implicated in the initial step of regeneration. In amphibian regeneration, the up-regulation of proteinases such as metalloproteinases (MMPs) and cathepsin D, and proteinase-related proteins such as proteinase tissue inhibitors and activators has been demonstrated. Since the earthworm could provide a unique and valuable model to investigate the mechanism of regeneration, we studied the developmental change in proteinase expression during earthworm tail regeneration. Zymographic analysis revealed that proteinase activities began to increase within 1 h after amputation and reached a maximum at 7 days post-amputation. This peak in activity was approximately 22-fold greater than the unamputated controls. Thereafter, the proteinase activities tended to decrease followed by another peak at 30 days before returning to control levels. At least four types of proteinase were distinguishable at 7 and 30 days post-amputation, with molecular weights of 25, 28, 38, and 44 kDa, respectively. All proteinase activities were strongly inhibited by addition of phenylmethylsulfonyl fluoride (PMSF) and aprotinin, specific inhibitors for serine proteinase. Pepstatin A, E-64, iodoacetamide and a metal ion-free medium were not effective inhibitors, indicating that proteinases expressed during earthworm tail regeneration would be serine proteinases. In addition, we were able to detect two types of plasminogen activator (PA) with molecular weights of 40 and 47 kDa, respectively. PA activities were predominantly expressed at 1, 5, and 25 days post-amputation, which preceded two peaks of serine proteinase activities appearing at approximately 7 and 30 days after amputation, respectively. This fact supports the view that serine proteinases expressed in respond to tail amputation may be plasmin-like proteinases activated by PA.     

Noninvasive diagnostics of bladder cancer based on analysis of telomerase activity and expression hTERT and hTR coding its subunits

In order to develop noninvasive diagnostics of bladder cancer (BC), telomerase activity has been examined by means the TRAP method (telomerase repeat amplification protocol) in tumor tissue and urine pellet samples taken from patients with bladder cancer. The levels of relative expression of genes encoding telomerase catalytic subunit (hTERT) and its RNA subunit (hTR) were evaluated by RT-PCR. Telomerase activity and expression of genes encoding its subunits were detected in both tumor tissues and in the urine cell pellet from each BC patient. Results of our study demonstrate possibility of noninvasive BC diagnostics using combination of these methods with sensitivity of 96% and specificity of 100% in the case of telomerase detection and with sensitivity of 80% and specificity of 100% in the case of hTERT detection in urine pellet samples.     

Effects of elevated CO2 and temperature on leaf characteristics,photosynthesis and carbon storage in aboveground biomass of a boreal bioenergy crop (Phalaris arundinacea L.) under varying water regimes

This work examined the effects of elevated CO₂ and temperature and water regimes, alone and in interaction, on the leaf characteristics [leaf area (LA), specific leaf weight (SLW), leaf nitrogen content (N_L) based on LA], photosynthesis (light‐saturated net carbon fixation rate, P_sat) and carbon storage in aboveground biomass of leaves (C_l) and stem (C_s) for a perennial reed canary grass (Phalaris arundinacea L., Finnish local cultivar). For this purpose, plants were grown under different water regimes (ranging from high to low soil moisture) in climate‐controlled growth chambers under the elevated CO₂ and/or temperature (following a factorial design) over a whole growing season (May–September in 2009). The results showed that the elevated temperature increased the leaf growth, photosynthesis and carbon storage of aboveground biomass the most in the early growing periods, compared with ambient temperature. However, the plant growth declined rapidly thereafter with a lower carbon storage at the end of growing season. This was related to the accelerated phenology regulation and consequent earlier growth senescence. Consequently, the elevation of CO₂ increased the P_sat, LA and SLW during the growing season, with a significant concurrent increase in the carbon storage in aboveground biomass. Low soil moisture decreased the P_sat, leaf stomatal conductance, LA and carbon storage in above ground biomass compared with high and normal soil moisture. This water stress effect was the largest under the elevated temperature. The elevated CO₂ partially mitigated the adverse effects of high temperature and low soil moisture. However, the combination of elevated temperature and CO₂ did not significantly increase the carbon storage in aboveground biomass of the plants.