Shelf-life extension of refrigerated Mediterranean mullet (Mullus surmuletus) using modified atmosphere packaging

The present work evaluated the quality and freshness characteristics and the effect of modified atmosphere packaging (MAP) on the shelf-life extension of refrigerated Mediterranean mullet using microbiological, biochemical, and sensory analyses. Fresh open sea red mullet (Mullus surmuletus) were packaged in four different atmospheres: M1, 10%/20%/70% (O2/ CO2/N2); M2, 10%/40%/50% (O2/CO2/N2); M3, 10%/60%/30% (O2/CO2/N2); identical fish samples were packaged in air. All fish were kept under refrigeration (4 +/- 0.5 degrees C) for 14 days. Of the three gas atmospheres, the 10%/40%/50% (M2) and 10%/ 60%/30% (M3) gas mixtures were the most effective for inhibiting growth of aerobic microflora in mullet samples until day 10 of refrigerated storage. H2S-producing bacteria and pseudomonads were part of the mullet microflora and their growth was partly inhibited under MAP conditions. Between these two bacterial groups, H2S-producing bacteria (including Shewanella putrefaciens) were dominant toward the end of the storage period, regardless of the packaging conditions. Brochothrix thermosphacta and lactic acid bacteria were found to be members of the final microbial flora of MAP and air-packaged mullet, whereas the Enterobacteriaceae population was lower than other bacterial groups. Of the chemical freshness indices determined, thiobarbituric acid values were variable in mullet samples irrespective of packaging conditions indicative of no specific oxidative rancidity trend. Based on sensorial data and aerobic plate count, trimethylamine nitrogen and total volatile basic nitrogen limit values in the range of ca. 15 to 23 and 52 to 60 mg N/100 g of fish muscle were obtained, respectively, for mullet packaged under modified atmosphere and air. Sensory analyses (odor and taste attributes) showed that the limit of sensorial acceptability was reached after ca. 6 days for the samples packaged in air, 8 days for the M1 and M3 samples, and after 10 days for the M2 samples. Respective shelf-life extension for fresh whole mullet was ca. 2 days (M1 and M3 gas mixtures), and 4 days (M2 gas mixture).     

Evaluation of contaminant removal of reverse osmosis and advanced oxidation in full-scale operation by combining passive sampling with chemical analysis and bioanalytical tools

Advanced water treatment of secondary treated effluent requires stringent quality control to achieve a water quality suitable for augmenting drinking water supplies. The removal of micropollutants such as pesticides, industrial chemicals, endocrine disrupting chemicals (EDC), pharmaceuticals, and personal care products (PPCP) is paramount. As the concentrations of individual contaminants are typically low, frequent analytical screening is both laborious and costly. We propose and validate an approach for continuous monitoring by applying passive sampling with Empore disks in vessels that were designed to slow down the water flow, and thus uptake kinetics, and ensure that the uptake is only marginally dependent on the chemicals' physicochemical properties over a relatively narrow molecular size range. This design not only assured integrative sampling over 27 days for a broad range of chemicals but also permitted the use of a suite of bioanalytical tools as sum parameters, representative of mixtures of chemicals with a common mode of toxic action. Bioassays proved to be more sensitive than chemical analysis to assess the removal of organic micropollutants by reverse osmosis, followed by UV/H?O? treatment, as many individual compounds fell below the quantification limit of chemical analysis, yet still contributed to the observed mixture toxicity. Nonetheless in several cases, the responses in the bioassays were also below their quantification limits and therefore only three bioassays were evaluated here, representing nonspecific toxicity and two specific end points for estrogenicity and photosynthesis inhibition. Chemical analytical techniques were able to quantify 32 pesticides, 62 PCPPs, and 12 EDCs in reverse osmosis concentrate. However, these chemicals could explain only 1% of the nonspecific toxicity in the Microtox assay in the reverse osmosis concentrate and 0.0025% in the treated water. Likewise only 1% of the estrogenic effect in the E-SCREEN could be explained by the quantified EDCs after reverse osmosis. In comparison, >50% of the estrogenic effect can typically be explained in sewage. Herbicidal activity could be fully explained by chemical analysis as the sampling period coincided with an illegal discharge and two herbicides dominated the mixture effect. The mass balance of the reverse osmosis process matched theoretical expectations for both chemical analysis and bioanalytical tools. Overall the investigated treatment train removed >97% estrogenicity, >99% herbicidal activity, and >96% baseline toxicity, confirming the suitability of the treatment train for polishing water for indirect potable reuse. The product water was indistinguishable from local tap water in all three bioassays. This study demonstrates the suitability and robustness of passive sampling linked with bioanalytical tools for semicontinuous monitoring of advanced water treatment with respect to micropollutant removal.     

Negative electrospray droplet exposure to gaseous bases for the manipulation of protein charge state distributions

The exposure of electrospray droplets to vapors of reagents of various base strengths affects protein negative charge state distributions independent of initial solution conditions. Volatile bases are introduced into the counter-current nitrogen drying gas of an electrospray interface to interact with charged droplets as they undergo desolvation/disintegration, shifting charge state distributions of proteins to higher, more negative, charge states. Alterations of charge state distributions can implicate protein folding/unfolding phenomena. Species bound by relatively weak interactions can be preserved, at least to some extent, allowing for the observation of high charge states of protein-ligand complexes, such as high negative charge states of holomyoglobin. The binding of carbonic anhydrase with its Zn(2+) cofactor is apparently preserved when the holo-form of the protein is exposed to basic vapors (i.e., the Zn(2+) ion remains associated with the protein), but this prevents the appearance of charge states higher than -17. Charge state distributions of proteins containing disulfide bonds shift slightly with the leak-in of basic vapors, but when these disulfide bonds are reduced with dithiothreitol in solution, charge states higher than the number of acidic sites (Asp, Glu, and C-terminus) are observed. Since there is no observed change in the distributions of buffered proteins exposed to these reagent vapors, the charge state changes are attributed largely to a pH affect. High pK(a) and highly volatile reagents have been found to be the most effective in terms of observing the maximum negative charge state of the biomolecule of interest.     

The Effects of Acidosis on eNOS in the Systemic Vasculature: A Focus on Early Postnatal Ontogenesis

The activity of many vasomotor signaling pathways strongly depends on extracellular/intracellular pH. Nitric oxide (NO) is one of the most important vasodilators produced by the endothelium. In this review, we present evidence that in most vascular beds of mature mammalian organisms metabolic or respiratory acidosis increases functional endothelial NO-synthase (eNOS) activity, despite the observation that direct effects of low pH on eNOS enzymatic activity are inhibitory. This can be explained by the fact that acidosis increases the activity of signaling pathways that positively regulate eNOS activity. The role of NO in the regulation of vascular tone is greater in early postnatal ontogenesis compared to adulthood. Importantly, in early postnatal ontogenesis acidosis also augments functional eNOS activity and its contribution to the regulation of arterial contractility. Therefore, the effect of acidosis on total peripheral resistance in neonates may be stronger than in adults and can be one of the reasons for an undesirable decrease in blood pressure during neonatal asphyxia. The latter, however, should be proven in future studies.     

Capsule-Targeting Depolymerases Derived from Acinetobacter baumannii Prophage Regions

In this study, several different depolymerases encoded in the prophage regions of Acinetobacter baumannii genomes have been bioinformatically predicted and recombinantly produced. The identified depolymerases possessed multi-domain structures and were identical or closely homologous to various proteins encoded in other A. baumannii genomes. This means that prophage-derived depolymerases are widespread, and different bacterial genomes can be the source of proteins with polysaccharide-degrading activities. For two depolymerases, the specificity to capsular polysaccharides (CPSs) of A. baumannii belonging to K1 and K92 capsular types (K types) was determined. The data obtained showed that the prophage-derived depolymerases were glycosidases that cleaved the A. baumannii CPSs by the hydrolytic mechanism to yield monomers and oligomers of the K units. The recombinant proteins with established enzymatic activity significantly reduced the mortality of Galleria mellonella larvae infected with A. baumannii of K1 and K92 capsular types. Therefore, these enzymes can be considered as suitable candidates for the development of new antibacterials against corresponding A. baumannii K types.     

Killing for museums: European bison as a museum exhibit

The European bison is one of the last remnants of the megafauna that once roamed through Europe. By the early modern period, it had already disappeared from most of its former range and had become a coveted natural curiosity as well as been designated as royal game. In the 18th century, the last population of lowland European bison surviving in the Bia?owie?a Forest became an object of study for naturalists. When the forest became a part of the Russian Empire during the 19th and early 20th centuries, it was the Russian Imperial family's attention to the species that ensured its survival. In the same period, the European bison was of the utmost importance to the scientific community—as a research subject (on the taxonomic position of the species, its relation to extinct Bovidae, etc.) and as a museum exhibit (to attract audiences interested in seeing the largest European mammal). Obtaining such a specimen demanded the scientific community's adept maneuvering through diplomatic and bureaucratic channels and depended on the Russian monarch's goodwill. The tsar's permission was only a start, as naturalists themselves had to organize hunts, process the pelt and bones, and deliver them to museums. Despite the considerable organizational and logistical difficulties, by the late 19th and early 20th centuries, many museums were displaying the European bison, which succeeded in making the species well known to the wider public. In 1919, the last wild Bia?owie?a bison was killed, but the popularity of this species helped the reintroduction of the animal a decade later. Today, the historical specimens are a source of samples for both genetic and molecular research.