Oxidative injury in V79 Chinese hamster cells: protective role of the superoxide dismutase mimetic MnTM-4-PyP

Oxidative cell injury could be induced by different reactive oxygen species (ROS) operating in multiple pathways. The present work is focused on three different models of oxidative stress: the xanthine/xanthine oxidase system (XXO), an extracellular superoxide anion generator; tert-butylhydroperoxide (TBHP), an analogue of lipid hydroperoxides; and doxorubicin (Dox), an anticancer drug. Superoxide and peroxyl radicals, among other ROS, could be effectively scavenged by MnTM-4-PyP, a polyfunctional catalytic antioxidant. In this report, we have addressed the role of MnTM-4-PyP on the protection against the cytotoxicity induced by the three aforementioned oxidants. The effect of MnTM-4-PyP (0.1–100 μM) was evaluated in V79 fibroblasts using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide reduction and the crystal violet assays, as well as the mitotic index. Also, the generation of intracellular ROS was studied by the fluorescent probe dihydroethidium. MnTM-4-PyP has shown significant protective effects against the cytotoxicity of XXO and TBHP, increasing the cell viability in approximately 40% and reducing the intracellular level of ROS. However, no considerable protection occurred against Dox. The three oxidants caused a mitotic index reduction that was not altered by MnTM-4-PyP. In summary, MnTM-4-PyP appears to be a promising agent for the protection against oxidative injury. However, it has shown differential responses, reinforcing the need to study different experimental models for the adequate evaluation of its potentialities as a catalytic antioxidant.     

A radioassay for phosphofructokinase-1 activity in cell extracts and purified enzyme

Phosphofructokinase-1 plays a key role in the regulation of carbohydrate metabolism. Its activity can be used as an indicator of the glycolytic flux in a tissue sample. The method most commonly employed to determine phosphofructokinase-1 activity is based on oxidation of NADH by the use of aldolase, triosephosphate isomerase, and alpha-glycerophosphate dehydrogenase. This method suffers from several disadvantages, including interactions of the auxiliary enzymes with phosphofructokinase-1. Other methods that have been used also require auxiliary enzymes or are less sensitive than a coupled assay. Here, we propose a direct method to determine phosphofructokinase-1 activity, without the use of auxiliary enzymes. This method employs fructose-6-phosphate and ATP labeled with 32P in the gamma position ([gamma-32P]ATP), and leads to the formation of ADP and fructose-1,6-bisphosphate labeled with 32P ([1-32P]fructose-1,6-bisphosphate). Activated charcoal is used to adsorb unreacted [gamma-32P]ATP, and the radioactive product in the supernatant, [1-32P]fructose-1,6-bisphosphate, is analyzed on a liquid scintillation counter. The proposed method is precise and relatively inexpensive, and can be applied to determine phosphofructokinase-1 activity in cellular extracts as well as in the purified enzyme.     

On the phylogeny of Ischyroceridae (Amphipoda,Senticaudata, Corophiida), with the description of a new genus and eight new species from deep‐sea Brazilian waters

The family Ischyroceridae is analysed herein by cladistic methods based on morphological characters, using both PAUP 4.0b and TNT. The data matrix of 41 characters × 32 terminal taxa was constructed using DELTA. Based on the results, we comment on the phylogenetic relationships of certain genera and their synapomorphic characters, also discussing the phylogenetic position of M yersius gen. nov. , which appeared as the sister group of Bathyphotis. In addition, Pseudischyrocerus crenatipes is removed to Bathyphotis, for which a new diagnosis is provided. A taxonomic study with the Ischyroceridae collected on the continental slope (depth, 700–2000 m) in the Campos Basin (20.5–24°S, 40–41°W) was also performed. Samples were collected in November–December 2002 and July–August 2003 using a box core device. As a result, a new genus and eight new species are described: B onnierella campensis sp. nov. , B onnierella laurensi sp. nov. , M yersius denticaudatus gen. et sp. nov. , N otopoma lowryi sp. nov. , N otopoma teresae sp. nov. , P seudericthonius bousfieldi sp. nov. , P seudericthonius concavus sp. nov. , and P seudischyrocerus caecus sp. nov. The genus Bonnierella is recorded for the first time from Brazilian waters, and the subspecies Bonnierella linearis linearis and Bonnierella linearis californica are elevated to species rank. Keys to the genera of Ischyroceridae used in the cladistic analysis and the world species of Notopoma are given. © 2014 The Linnean Society of London     

Combined evaluation of bovine tuberculosis in wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>) and red deer (<Emphasis Type="Italic">Cervus elaphus</Emphasis>) from Central-East Portugal

Bovine tuberculosis (bTB), widely considered a disease of livestock, can also affect large game species despite their poorly understood role in the epidemiology of the endemic state of the disease. The Idanha-a-Nova County, situated in the eastern region of central Portugal, is one of the most important regions for large game hunting activity. In order to investigate the role of large game and the presence of bTB in Idanha-a-Nova, 132 wild boar (Sus scrofa) and 339 red deer (Cervus elaphus) were inspected for bTB-compatible lesions (bTBCL) during the 2008–2009 hunting season. Tissue samples with bTBCL were collected and processed for bacteriological confirmation and spoligotyping of Mycobacterium bovis isolates. In wild boar, bTB lesions were single, mostly calcified (21/29, 72.4%) and situated in the mandibular lymph nodes. Lesions were more severe and largely non-calcified (36/40, 90%) in red deer. Wild boar presented a consistently higher bTB infection rate than the red deer; however, the difference was only significant in the southern B area. This area was significantly affected by bTB for both animal species (up to 72.7% in wild boar and 34.8% in red deer) and requires the urgent implementation of intervention measures to contain the disease. Wild boar may assume an important role in the emergence of new disease foci in distant areas that infected red deer cannot reach. Spoligotyping of M. bovis isolates grouped the strains into three clusters, revealing patterns common to wild boar, red deer, and also to local cattle, indicating possible transmission among different animal species.     

Evaluation of domestic wastewater treatment using microalgal-bacterial processes: effect of CO<Subscript>2</Subscript> addition on pathogen removal

Microalgal-bacterial processes represent a sustainable and cost-effective biotechnology able to promote efficient wastewater treatment, including natural pathogen removal (disinfection), as well as being able to perform CO₂ uptake and biogas upgrading. In this context, the influence of CO₂ supply from a synthetic gas mixture (30% v/v CO₂) on the removal of pathogens (Pseudomonas, enterococci, and Escherichia coli) and total coliforms during secondary domestic wastewater treatment by a microalgal-bacterial symbiosis in a 180-L high-rate algal pond (HRAP) was investigated. The supply of CO₂ in the HRAP positively influenced the Pseudomonas aeruginosa removal, with the removal efficiency increasing from 97.4% (1.6 log) to 99.6% (2.5 log) without and with CO₂ supply, respectively. Likewise, the total coliform removal efficiency rose from 88.7% (1.1 log) to 99.4% (2.8 log). On the other hand, the effect of CO₂ supply on enterococci (99.7% and 2.6 log) and Escherichia coli (98.6% and 2.2 log) removal was negligible.     

Life history told by a whale-louse: a possible interaction of a southern right whale <Emphasis Type="Italic">Eubalaena australis</Emphasis> calf with humpback whales <Emphasis Type="Italic">Megaptera novaeangliae</Emphasis>

Southern right whales (Eubalaena australis) are known to host three species of whale-lice, Cyamus gracilis, Cyamus ovalis and Cyamus erraticus. Such cyamids usually are generalists in toothed whales (Cetacea: Odontoceti) and host-specific in baleen whales (Cetacea: Mysticeti), and because they have no free-swimming stage, transmission only occurs by contact between whales. One southern right whale stranded at the southeastern coast of Brazil was found parasitized by a different species of cyamid. Over 300 specimens were collected and the only species identified based on morphological and molecular data was Cyamus boopis, a typical ectoparasite of humpback whales (Megaptera novaeangliae). This finding is the first record of C. boopis on the southern right whale. Both E. australis and M. novaeangliae are found in Brazilian waters and the presence of humpback’s whale-louse together with the lack of the three specific parasites of right whales suggest an interspecific interaction between these whales based on the parasite’s biology.     

Cladistic revision of talitroidean amphipods (Crustacea, Gammaridea), with a proposal of a new classification

This paper reports the results of a cladistic analysis of the Talitroidea s.l. , which includes about 400 species, in 96 genera distributed in 10 families. The analysis was performed using paup and was based on a character matrix of 34 terminal taxa and 43 morphological characters. Four most parsimonious trees were obtained with 175 steps (CI = 0.617, RI = 0.736). A strict consensus tree was calculated and the following general conclusions were reached. The superfamily Talitroidea is elevated herein as infraorder Talitrida, which is subdivided into three main branches: a small clade formed by Kuria and Micropythia (the Kurioidea), and two larger groups maintained as distinct superfamilies (Phliantoidea, including six families, and Talitroidea s.s ., including four). Within the Talitroidea s.s. , the following taxonomic changes are proposed: Hyalellidae and Najnidae are synonymized with Dogielinotidae, and treated as subfamilies; a new family rank is proposed for the Chiltoniinae.     

Macaronesian islands as promoters of diversification in amphipods: The remarkable case of the family Hyalidae (Crustacea,Amphipoda)

The north‐east Atlantic (NEA) is an important and complex biogeographic region with a very rich marine fauna. However, little is known about the role of the Macaronesian islands in the evolutionary history and diversification of marine invertebrates in the NEA. Among the amphipods, the members of the family Hyalidae are particularly common and abundant in intertidal rocky shores of NEA. In this study, we aimed to investigate the genetic structure and diversity of seven hyalid species inhabiting the Macaronesian, European and Moroccan Atlantic coasts, with a focus on the genetic differentiation between island and Continental populations. Analysis of mitochondrial DNA sequences of the cytochrome oxidase I gene, unravelled a very high level of hidden diversity, consisting of 26–32 molecular operational taxonomic units (MOTUs), the majority of them recorded in Macaronesian populations. Except for Apohyale stebbingi, all remaining MOTUs were in general allopatrically distributed, with a trend for segregation between islands and Continental populations on one side, but also for the occurrence of private MOTUs among islands. Results indicate distinct evolutionary and diversification patterns among Hyalidae species, but a strong separation between Continental and islands’ lineages appears to be a common feature to all of them. Apparently, the complex geomorphological history of the Macaronesian archipelagos served as an important promoter of extensive diversification of marine invertebrates in NEA, a phenomenon which only now starts to be fully appreciated through the use of molecular data.