Mass spectrometry of intact V-type ATPases reveals bound lipids and the effects of nucleotide binding

The ability of electrospray to propel large viruses into a mass spectrometer is established and is rationalized by analogy to the atmospheric transmission of the common cold. Much less clear is the fate of membrane-embedded molecular machines in the gas phase. Here we show that rotary adenosine triphosphatases (ATPases)/synthases from Thermus thermophilus and Enterococcus hirae can be maintained intact with membrane and soluble subunit interactions preserved in vacuum. Mass spectra reveal subunit stoichiometries and the identity of tightly bound lipids within the membrane rotors. Moreover, subcomplexes formed in solution and gas phases reveal the regulatory effects of nucleotide binding on both ATP hydrolysis and proton translocation. Consequently, we can link specific lipid and nucleotide binding with distinct regulatory roles.     

Dynamics of public attitudes toward bears and the role of bear hunting in Croatia

Successful carnivore conservation depends on public attitudes and acceptance levels of carnivores, and these are likely to change as circumstances change. Attitude studies repeated in time that can demonstrate such change are rare. Our study surveyed Croatian rural inhabitants in 2002 and in 2008 and analyzed their responses to detect a change in attitudes toward brown bears (Ursus arctos) over time. Important developments occurring in Croatia at the time of our research included a more centralized and more clearly defined bear management strategy, and an increase in the bear population. We constructed models to explain respondent’s value orientations, their level of perceived threat and their acceptance capacity for bears. Findings show that while value orientations and the overall level of perceived threat did not change over time, bear acceptance capacity was reduced. This suggests that the increase in the bear population and perhaps the more centralized bear management reduced respondents’ willingness to accept a larger bear population. We conclude that continuous public involvement in bear management is essential in order to maintain a feeling of control over the bear among the local population. Furthermore we argue that hunting is an important form of public involvement in the region, serving to reinforce existence and bequest values of the bear and increase its public acceptance.     

Organic-geochemical Differentiation of Petroleum-type Pollutants and Study of Their Fate in Danube Alluvial Sediments and Corresponding Water (Pančevo Oil Refinery, Serbia)

A review is given in this paper of the up-to-date results observed in differentiation and transformation studies on petroleum-type pollutants in underground and surface waters. Water and particulate matter derived from the locality of Pan?evo Petroleum Refinery, Serbia (River Danube alluvial formations). It was shown that distributions of n-alkanes, steranes and triterpanes, and δ¹³C_PDB values of n-alkanes may successfully be used for qualitatively differentiating the petroleum-type pollutants from native organic matter in recent sedimentary formations. In underground waters, a petroleum-type pollutant is exposed to microbiological degradation which is manifested through relatively fast degradation of n-alkanes. Following an almost complete degradation of crude oil n-alkanes in underground water, the biosynthesis of novel, even carbon-number C₁₆–C₃₀ n-alkanes may be observed. It is shown that the n-alkane distribution observed in a petroleum-type pollutant may depend on the intensity of its previous interaction with water. The fate of petroleum-type pollutants in environmental waters may be predicted through laboratory simulative microbiological degradation experiments by using microorganism consortiums similar to those observed under relevant natural conditions, as well as on corresponding nutrient base.     

Spatial and vertical distribution of <Superscript>137</Superscript>Cs in soils in the erosive area of southeastern Serbia (Pčinja and South Morava River Basins)

Purpose

The area of southeastern Serbia, the P?inja and South Morava River Basins, is under the influence of very strong erosion, and the aim of this study was to investigate the vertical and spatial distribution of the ¹³⁷Cs in the eroded soils of this area.

Materials and methods

Vertical soil profiles were collected with 5-cm increments from the uppermost layer down to 20, 25, 30, 40, and 50 cm of depth, depending on the thickness of the soil layers, i.e., down to the underlying parent rocks. Measurements of ¹³⁷Cs activity concentration were performed by using the HPGe gamma-ray spectrometer ORTEC-AMETEK (34 % relative efficiency and high resolution 1.65 keV at 1.33 MeV for ⁶⁰Co), from its gamma-ray line at 661.2 keV.

Results and discussion

The mean ¹³⁷Cs activity concentration across all 18 soil profiles (for all soil layers) was found to be 20 Bq kg^?1. In the greatest number of soil profiles, the ¹³⁷Cs activity concentration was generally highest in the first soil layer (0–5 cm) and decreased with soil depth, while in a few soil profiles, the peak of either the ¹³⁷Cs activity concentration occurred in the second soil layer (5–10 cm) or the ¹³⁷Cs activity concentration was almost equal throughout the entire soil profile. The mean ¹³⁷Cs activity concentration in the first soil layer (0–5 cm) was found to be 61 Bq kg^?1, and the high coefficient of variation of 92 % pointed out high spatial variability and large range of the ¹³⁷Cs activity concentrations in the study area.

Conclusions

The obtained results indicate that in the greatest number of soil profiles, ¹³⁷Cs is present in the upper layers, with concentration decreasing with depth, as is typical in uncultivated soil. Its spatial distribution was very uneven among the surface soil layers of the investigated sites. One of the main reasons for such pattern of ¹³⁷Cs in the study area may be soil erosion. Additional investigations which would support this hypothesis are required.

     

Antimutagenic effects of extracts from sage (Salvia officinalis) in mammalian system in vivo

Naturally occurring antimutagenic factors, especially those of plant origin, have recently become a subject of intensive research. Antimutagenic properties of terpenoid fractions of sage (Salvia officinalis) were tested in mammalian system in vivo through examining the ability of sage to decrease the frequency of aberrant cells induced by a potent mutagen. First, groups of mice were treated with three concentrations of sage alone and it was established that the frequency of aberrant cells after treatment with a concentration of 25 microL/kg was not significantly different from the negative control (olive oil), while that found after treatment with the 50 microL/kg concentration differed significantly (chi2(1) = 4.05, p < 0.05). Sage used at a concentration of 100 microL/kg was cytotoxic. Mitomycin C (MMC), known as a potent mutagen, was used for induction of chromosome aberrations. Post-treatment with sage suppressed the effects of MMC significantly. Both concentrations (25 microL/kg and 50 microL/kg) produced a significant decrease in the frequency of aberrations relative to MMC (chi2(1) = 5.42, p < 0.02, chi2(1) = 14.93, p < 0.001, respectively). The percent of aberrations decreased with increasing concentrations of sage. Only nontoxic concentrations of sage without mutagenic effects can be recommended for use as inhibitors of mutagenesis or carcinogenesis.     

Comparative analysis of using cereal grains and compound feed in semi-intensive common carp pond production

The semi-intensive system of carp farming is dominant in inland aquaculture. Although it appears to be simple, this system has a whole range of options, from those based on traditional rearing methods with use of plants from the region where the fish are reared to production based on the use of high-quality concentrated feeds. In this paper, we conduct a comparative analysis of the effect of using cereal grains as opposed to compound feed (pelleted and extruded) on the quantity of fish produced, flesh quality, histological changes in the digestive tract, water quality, and structure of the biocoenosis in the pond ecosystem. When the fish are fed cereal grains, production ranges from 500 to 1500 kg/ha, while using compound feed gives production that is twice as great. The proportions of protein and water are higher and that of lipids lower in flesh of common carp reared on compound feed in relation to fish fed cereal grains. Use of extruded feed gives better content of n-3 polyunsaturated fatty acids and a better n-3/n-6 ratio than when pelleted feed and cereal grains are used. Comparison of water quality in carp ponds under conditions of using various types of feed indicates that there are no significant differences. Although it was expected that feed with a higher percentage of proteins would cause disturbances in the aquatic ecosystem, their continual incorporation into the cycling of matter in the water contributes to maintenance of ecological equilibrium and even indirectly promotes growth of the common carp.     

Distribution and Genotypic Characterization of Campylobacter jejuni Isolated from Poultry in Split and Dalmatia County,Croatia

Consumption of poultry contaminated with Campylobacter jejuni has been recognized worldwide as the leading cause of campylobacteriosis. Therefore, the aim of our study was to investigate the prevalence and genotype diversity of Campylobacter jejuni in poultry meat intended for consumption in Split and Dalmatia County, which is the second biggest County in Croatia. Furthermore, we also wanted to discover possibly stable clones of C. jejuni appearing in different samples and periods of time, which would indicate their ability to persist in or adapt to poultry. In the period from March 2008 until June 2010, 834 samples of poultry from various sources were examined using a surface swab technique. Isolation of C. jejuni was performed by Preston broth and Karmali agar. Identification of the isolates was carried out using biochemical tests. C. jejuni was found in 84 of 574 chicken samples (14.6%) and in nine of 260 samples of turkey (3.5%). Pulse‐field gel electrophoresis (PFGE) was used to analyse 61 obtained isolates using SmaI and KpnI. Of 22 different macrorestriction profiles (MRP) that were found, five were detected in poultry from both different locations and periods of time. Samples from 11 locations were found to be contaminated with more than two different genotypes of C. jejuni. Interestingly, the same MRP were found both in poultry declared to be of domestic origin and in the poultry imported from abroad. The prevalence of C. jejuni in poultry samples was in accordance with previously reported results. Genotypic analysis indicated that the population of C. jejuni in Split and Dalmatia County was diverse and that multiple strains of C. jejuni could be found in the same poultry samples. Furthermore, the same genotypes were identified from the samples obtained from different locations and periods of time, which could support the theory of a global existence of certain MRP that are able to persist in or adapt to poultry.     

Fructose 1,6-biphosphate aldolase in the sera of Simmental young bulls Connection with the fattening capacity, muscle quantity and protein content

SUMMARY: The activity of enzyme aldolase (ALD) was determined in the sera of Simmental young bulls, with the purpose to see whether there exists a relationship between the enzyme activity and fattening capacity and whether the ALD could be an indicator of the muscle quantity and protein content. Early ALD activity was correlated with slaughter weight and ADG in the last fattening month. The regression analysis suggests that young bulls with higher serum ALD acitivity at the start of the experiment had more total muscles and less bone in the analysed rib part. On the basis of ALD activity in the first fattening month it was possible to estimate protein and fat content in the MLD. ZUSAMMENFASSUNG: Fructose-1,6-biphosphat Aldolase beim Serum Simmentaler Jungbullen. Beziehung zwischen Mastf?higkeit, Muskelmenge und Proteingehalt Aldolase (ALD) Enzymt?tigkeit im Serum von Simmentaler Jungbullen wurden in Hinblick auf eine Beziehung mit Mastf?higkeit, Muskelmenge und Proteingehalt untersucht. Es zeigt sich eine Korrelation zwischen früher ALD Aktivit?t und Schlachtgewicht und Tageszuwachs im Endmastmonat. Regressionsanalyse ergab, da? Jungbullen mit h?herer ALD Aktivit?t bei Mastbeginn mehr Muskelanteil im m. longissimus dorsi und weniger Knochen besitzen. ALD Aktivit?t im ersten Mastmonat erlaubt Sch?tzung des Protein- und Fettgehalts im m. long, dorsi. RESUMEN: Fructosa 1,6-bifosfato aldolasa en el suero de los terneros simentales. Relatión con sus capacidades de ser cebados, su cantidad de músculos y el contenido de proteínas Con el fin de establecer la relación entre la actividad de enzimas y las capacidades durante el proceso de la ceba, asf como para precisar si el ALD puede servir como indicador de la cantidad de músculos y el contenido de proteinas, intentaba determinarse la actividad de los enzimas de aldolasa (ALD) en el suero de los terneros simentales. A base de la actividad indicial del ALD, es posible estimar el peso final de matadero y la crecencia diaria media durante el último mes de la ceba. Tras una análisis regresiva se ha probado que los terneros con una actividad mayor del ALD en el suero a principios del proceso de su ceba tienen más músculos en total en la parte de costillas y m. longissimus dorsi mientras que tienen menos huesos. A base de la actividad del ALD durante el primer mes de la ceba es posible estimar el contenido de proteinas y grasa en m. long, dorsi. RéSUMé: Fructose 1.6-biphosphate aldolase dans le sérum des taurillons de Simmental. Le rapport entre l'aptitude à l'angraissement, la proportion des muscles et la teneur en protéines On a déterminé l'activité de l'enzyme aldolase (ALD) dans le sérum des taurillons de Simmental afin de vérifier s'il existait un rapport entre l'activité des enzymes et l'aptitude à l'engraissement et si l'ALD pouvait être indicatrice de la proportion de muscles et de la teneur en protéines. A la base de l'activité initiale de l'ALD, il est possible d'évaluer le poids de la carcasse et le gain quotidien moyen au cours du dernier mois d'engrais. Par analyse régressive, il a été établi que les taurillons à l'activité supérieure de l'ALD dans le sérum avaient au début de l'engrais plus de muscles au total dans la région costale et le m. longissimus dorsi et moins d'os. A la base de l'activité de l'ALD au cours du premier mois d'engrais il est possible d'évaluer la teneur en protéines et en graisse dans le muscle longissimus dorsi.     

Phosphorus-use efficiency of kale genotypes from coastal Croatia

Understanding the mechanisms of phosphorus (P)-use efficiency (PUE) may contribute to enhancing crop P nutrition because species growth variability at low-P is well known. The experiment was carried out to evaluate the response of kale genotypes to different P supply in randomized block design in three replications. Low-P supply led to a decrease in most parameters, whereas an increase was recorded in root growth parameters. Genotypes differed in shoot dry weight (DW), leaf area, root length and area, and shoot and root P content and concentration. Root traits significantly positively correlated with PUE. Genotypes Red Russian (RR) and IJK 17 were superior in terms of shoot DW production at low P supply, and had the highest uptake efficiency. Genotypes IJK 17 and 81 had the highest P utilization efficiency, while Vates blue curled (VBC) showed the lowest PUE. Genotypes had similar shoot P content and concentration at low P supply, but large PUE differences, implying the importance of P utilization efficiency.     

Oxidant/antioxidant properties of Croatian native propolis

Native propolis was defined as propolis powder collected from the continental part of Croatia and prepared according to a patented process that preserves all the propolis natural nutritional and organoleptic qualities. Nine phenolic compounds (out of thirteen tested) in propolis sample were detected by high performance liquid chromatography (HPLC) analysis. Among them chrysin was the most abundant (2478.5 microg/g propolis). Contrary to moderate antioxidant activity of propolis examined in vitro (ferric reduction antioxidant power; FRAP-assay), propolis as a food supplement modulated antioxidant enzymes (AOE) and significantly decreased lipid peroxidation processes (LPO) in plasma, liver, lungs, and brain of mice. The effect was dose- and tissue-dependent. The lower dose (100 mg/kg bw) protected plasma from oxidation, whereas the higher dose (300 mg/kg bw) was pro-oxidative. Hyperoxia (long-term normobaric 100% oxygen) increased LPO in all three organs tested. The highest vulnerability to oxidative stress was observed in lungs where hyperoxia was not associated with augmentation of AOE. Propolis protected lungs from hyperoxia by increased catalase (CAT) activity. This is of special importance for lungs since lungs of adult animals are highly vulnerable to oxidative stress because of their inability to augment AOE activity. Because of its strong antioxidant and scavenging abilities, native propolis might be used as a strong plant-based antioxidant effective not only in physiological conditions but also in cases that require prolonged high concentration of oxygen.