Research on the ability of propionic acid and vitamin B12 biosynthesis by <Emphasis Type="Italic">Propionibacterium freudenreichii</Emphasis> strain T82

The purpose of this study was to determine the potential for biosynthesis of propionic acid and vitamin B12 by Propionibacterium freudenreichii T82 in a medium containing various sources of carbon (glucose, fructose, and saccharose). These sugars are present in apple pomaces, which are the waste from the production of apple juice. Using statistical analysis design of experiments (DoE), the results allowed us to determine which sugars (carbon sources) exert the most beneficial influence on the biosynthesis of propionic acid and cobalamin. The highest production of propionic acid by the tested bacterial strain was obtained in a medium in which glucose accounted for at least 50% of the available carbon sources. Depending on the culture medium, the concentration of this metabolite ranged from 23 to 40 g/L. P. freudenreichii T82 produced the smallest amount of acid in medium in which the dominant nutrient source was saccharose. The results obtained indicated an inverse relationship between the amount of acid produced by the bacteria and vitamin B12 biosynthesis. Because of the high efficiency of propionic acid biosynthesis by P. freudenreichii T82, the prospect of using this strain to obtain propionate with the simultaneous disposal of waste materials (such as apple pomaces) which contain glucose and/or fructose is very promising.     

Engineering TaqII bifunctional endonuclease DNA recognition fidelity: the effect of a single amino acid substitution within the methyltransferase catalytic site

The aim of this study was to improve a useful molecular tool—TaqII restriction endonuclease-methyltransferase—by rational protein engineering, as well as to show an application of our novel method of restriction endonuclease activity modulation through a single amino acid change in the NPPY motif of methyltransferase. An amino acid change was introduced using site-directed mutagenesis into the taqIIRM gene. The mutated gene was expressed in Escherichia coli. The protein variant was purified and characterized. Previously, we described a TspGWI variant with an amino acid change in the methyltransferase motif IV. Here, we investigate a complex, pleiotropic effect of an analogous amino acid change on its homologue—TaqII. The methyltransferase activity is reduced, but not abolished, while TaqII restriction endonuclease can be reactivated by sinefungin, with an increased DNA recognition fidelity. The general method for engineering of the IIS/IIC/IIG restriction endonuclease activity/fidelity is developed along with the generation of an improved TaqII enzyme for biotechnological applications. A successful application of our novel strategy for restriction endonuclease activity/fidelity alteration, based on bioinformatics analyses, mutagenesis and the use of cofactor-analogue activity modulation, is presented.     

The influence of exposure to immunosuppressive treatment during pregnancy on renal function and rate of apoptosis in native kidneys of female Wistar rats

Pregnancy puts a significant additional strain on kidneys. The aim of our study was to investigate the impact of immunosuppressive drugs on changes in native kidneys in female Wistar rats after exposure during pregnancy. The study was conducted on 32 dams, subjected to immunosuppressive regimens commonly used in the therapy of human kidney transplant recipients (cyclosporine A, mycophenolate mofetil and prednisone; tacrolimus, mycophenolate mofetil and prednisone; cyclosporine A, everolimus and prednisone). The animals received drugs for 2 weeks before pregnancy and during 3 weeks of pregnancy. In all treated dams lower body weight (but not kidney mass) and alterations in serum sodium and chloride ions were found; serum creatinine concentration was increased in dams treated with cyclosporine A, everolimus and prednisone. All treatment groups of dams showed increased apoptosis in the distal tubules. In histological examination the changed intensity of acidophilic or basophilic cytoplasm of epithelial cells was found in kidneys of rats treated with calcineurin inhibitors, mycophenolate mofetil and prednisone. All immunosuppressive regimens caused abnormalities affecting nephron tubules. Regimens containing calcineurin inhibitors and mycophenolate mofetil caused higher rate of apoptosis and more pronounced histopathological changes. Regimen based on everolimus despite the lower rate of apoptosis in the proximal tubules and lower accumulation of kidney injury markers revealed higher serum creatinine concentration. Thus, interpretation which combination of drugs is better or worse for long-lasting functioning of kidneys in pregnant females requires further studies.     

The oxidizing power of the glutathione thiyl radical as measured by its electrode potential at physiological pH

The oxidizing power of the thiyl radical (GS*) produced on oxidation of glutathione (GSH) was determined as the mid-point electrode potential (reduction potential) of the one-electron couple E(m)(GS*,H+/GSH) in water, as a function of pH over the physiological range. The method involved measuring the equilibrium constants for electron-transfer equilibria with aniline or phenothiazine redox indicators of known electrode potential. Thiyl and indicator radicals were generated in microseconds by pulse radiolysis, and the position of equilibrium measured by fast kinetic spectrophotometry. The electrode potential E(m)(GS*,H+/GSH) showed the expected decrease by approximately 0.06 V/pH as pH was increased from approximately 6 to 8, reflecting thiol/thiolate dissociation and yielding a value of the reduction potential of GS*=0.92+/-0.03 V at pH 7.4. An apparently almost invariant potential between pH approximately 3 and 6, with potentials significantly lower than expected, is ascribed at least in part to errors arising from radical decay during the approach to the redox equilibrium and slow electron transfer of thiol compared to thiolate.     

Heterozygosity–fitness correlations in blue tit nestlings (<Emphasis Type="Italic">Cyanistis caeruleus</Emphasis>) under contrasting rearing conditions

Understanding the relation between genetic variation and fitness remains a key question in evolutionary biology. Although heterozygosity has been reported to correlate with many fitness-related traits, the strength of the heterozygosity–fitness correlations (HFCs) is usually weak and it is still difficult to assess the generality of these associations in natural populations. It has been suggested that HFCs may become meaningful only under particular environmental conditions. Moreover, existing evidence suggests that HFCs may also differ between sexes. The aim of this study was to investigate correlations between heterozygosity in neutral markers (microsatellites) and fitness-related traits in a natural population of blue tits (Cyanistes caeruleus). Additionally, we tested whether sex and environmental conditions may influence the magnitude and direction of HFCs. We found a positive relationship between heterozygosity and body mass of 14 days post-hatching nestlings, but only among females. Our results suggest that the correlation between heterozygosity and nestling body mass observed among female offspring could be attributed to within-brood effects. We failed to find any evidence that environmental conditions as simulated by brood size manipulation affect HFCs.     

Refugial debate: on small sites according to their function and capacity

The occurrence and location of long-term refugia determine the current patterns of biodiversity on Earth. The importance of the refugial debate is certain to increase in response to observed and expected species extinctions caused by climate change. Small areas where species survive outside their core range are important, as unique natural phenomena and model systems for observing the response of species to climate change. They can play a crucial role as potential sources for species recovery in the future or can act as progenitors of a new species. While most authors believe that sites connected with only long-term isolation should be included into the refugium concept, this approach can result in the loss of linkage between the ecological and evolutionary processes taking place during different phases of the species range dynamics. Moreover, the papers often interpret the nature of described phenomena in different ways. In response, the conceptual scheme given in our letter summarises the patterns which occur during species range shift. It proposes an equivalent scheme for small refugial sites according to their function and capacity, based on the relict species concept. This approach and proposed terminology is tested on the example of two plant species with different pattern of the long term range dynamics. Our paper highlights the importance of sites harboring ‘trailing-edge’ young relicts for the future long-term persistence of the species (as old relicts) under unfavorable regional conditions. By considering the age gradients of small refugial sites it is possible to reveal community interactions, species traits or genes that drive the responses of biota to climate changes.     

Accumulation of amino acids and phenolic compounds in biochemical plant responses to feeding of two different herbivorous arthropod pests

The study aimed at comparing the changes in the content of free amino acids, phenolic compounds and the activity of PAL and TAL caused by two piercing-sucking arthropods: the grape mealybug (Pseudococcus maritimus Ehrh.) and the two-spotted spider mite (Tetranychus urticae Koch) in the leaves of orchid and strawberry, respectively. The obtained results show that the amino acid content and the ratio of amino acids to phenolic compounds increased in both plant species infested by the mealybug and the mite. However, such response was weakly dependent on changes in activity of the analysed enzymes. The pest feeding affected accumulation of the phenolic compounds, since the induction of the PAL activity in mealybug-infested orchid leaves during the first 5 h of the experiment preceded the increase in phenolic compounds during the first week of insect feeding. Instead, the increased activity of TAL was accompanied by elevated levels of phenolic compounds in the leaves of strawberry infested by mites. Mechanisms of biochemical plant responses induced by infestation of the studied herbivorous arthropods are discussed.     

Sexual dimorphism,asymmetry, and the effect of reproduction on pelvis bone in the bank vole, <Emphasis Type="Italic">Myodes glareolus</Emphasis>

Sexual dimorphism in anatomical traits has been widely studied in animals. Although pelvis dimorphism was mostly studied in humans, it occurs also in many other mammalian species. Here, we investigated sexual dimorphism in the pelvis of the bank vole Myodes glareolus using individuals with known sex and reproductive status of females (parous vs nulliparous). The analyses revealed that the size and shape of pelvis differed significantly between sexes, as well as between nulliparous and parous females. In comparison with males, females had a significantly longer pelvis and pubis bones and a longer obturator foramen length, but a smaller pubis width. Interestingly, the difference between parous and nulliparous females resembles that between females and males: parous females had bigger pelvis, which probably resulted from changes during pregnancy and after birth. Left bones were on average larger than right ones, but the magnitude of directional asymmetry was not different between sex and reproduction group. Moreover, we noticed that fluctuating asymmetry of pelvis and pubis length was higher in females than in males and higher in parous than in nulliparous females, what is presumably associated with locomotor performance. A discriminant function analysis performed for the four bone size traits showed that the traits can be effectively used for a nearly perfect recognition of sexes and also a quite reliable recognition of the reproductive status of females.     

16S rDNA Pyrosequencing Analysis of Bacterial Community in Heavy Metals Polluted Soils

Soil contamination with heavy metals is a widespread problem, especially prominent on grounds lying in the vicinity of mines, smelters, and other industrial facilities. Many such areas are located in Southern Poland; they are polluted mainly with Pb, Zn, Cd, or Cu, and locally also with Cr. As for now, little is known about most bacterial species thriving in such soils and even less about a core bacterial community—a set of taxa common to polluted soils. Therefore, we wanted to answer the question if such a set could be found in samples differing physicochemically and phytosociologically. To answer the question, we analyzed bacterial communities in three soil samples contaminated with Pb and Zn and two contaminated with Cr and lower levels of Pb and Zn. The communities were assessed with 16S rRNA gene fragments pyrosequencing. It was found that the samples differed significantly and Zn decreased both diversity and species richness at species and family levels, while plant species richness did not correlate with bacterial diversity. In spite of the differences between the samples, they shared many operational taxonomic units (OTUs) and it was possible to delineate the core microbiome of our sample set. The core set of OTUs comprised members of such taxa as Sphingomonas, Candidatus Solibacter, or Flexibacter showing that particular genera might be shared among sites ~40 km distant.