The draft genome of Ciona intestinalis: Insights into chordate and vertebrate origins

The first chordates appear in the fossil record at the time of the Cambrian explosion, nearly 550 million years ago. The modern ascidian tadpole represents a plausible approximation to these ancestral chordates. To illuminate the origins of chordate and vertebrates, we generated a draft of the protein-coding portion of the genome of the most studied ascidian, Ciona intestinalis. The Ciona genome contains similar to16,000 protein-coding genes, similar to the number in other invertebrates, but only half that found in vertebrates. Vertebrate gene families are typically found in simplified form in Ciona, suggesting that ascidians contain the basic ancestral complement of genes involved in cell signaling and development. The ascidian genome has also acquired a number of lineage-specific innovations, including a group of genes engaged in cellulose metabolism that are related to those in bacteria and fungi.

The DMSO reductase family of microbial molybdenum enzymes; Molecular properties and role in the dissimilatory reduction of toxic elements

The dimethylsulfoxide (DMSO) reductase family of molybdenum enzymes is a large and diverse group that is found in bacteria and archaea. These enzymes are characterised by a bis(molybdopterin guanine dinucleotide)Mo form of the molybdenum cofactor, and they are particularly important in anaerobic respiration including the dissimilatory reduction of certain toxic oxoanions. The structural and phylogenetic relationship between the proteins of this family is discussed. High-resolution crystal structures of enzymes of the DMSO reductase family have revealed a high degree of similarity in tertiary structure. However, there is considerable variation in the structure of the molybdenum active site and it seems likely that these subtle but important differences lead to the great diversity of function seen in this family of enzymes. This diversity of catalytic capability is associated with several distinct pathways of electron transport.

Relationship between flocculation of activated sludge and composition of extracellular polymeric substances

Activated sludge floes are a flocculated mass of microorganisms, extracellular polymeric substances (EPS) and adsorbed organic and inorganic material. The structure of the floes is very heterogeneous and floes with very different properties and morphologies may occur, depending on the conditions in the activated sludge treatment plant and wastewater composition. Present thinking suggests that cations, such as calcium, create cationic bridges with EPS excreted by the bacteria and thereby hold the various floe constituents together. However, due to the complex and heterogeneous nature of activated sludge, the mechanisms have neither been thoroughly investigated nor successfully quantified. A better understanding and description of the biological flocculation process is necessary in order to establish more efficient operational strategies. The main aim of this study was to get a comprehensive and unique insight into the floe properties of activated sludge and to assess the relative impact of chemical and physical parameters. A variety of sludges from full scale treatment plants with different settling properties were characterised. The interrelationships between floe parameters such as composition of EPS, surface properties and floe structure, and their effect on the flocculation and separation properties were assessed. The results indicate that the EPS, both in terms of quantity and quality, are very important for the floe properties of the activated sludge. However, presence of filaments may alter the physical properties of the floes considerably. The EPS showed positive correlations to sludge volume index (SVI) if only sludges with low or moderate numbers of filaments were included. The surface properties were more affected by the composition of the EPS than by the number of filaments. The EPS showed positive correlation to negative surface charge and a negative correlation to relative hydrophobicity and flocculation ability. The negative correlation between flocculation ability and amount of EPS was surprising. The shear sensitivity, measured as degree of erosion of floes when subjected to shear, was more affected by floe size and number of filaments than amount of EPS.

Antibacterial activity of Baccharis trimera (Less.) DC. (carqueja) against bacteria of medical interest

Baccharis trimera (Less.) (Asteraceae), popularly know as "carqueja", is a species commonly used in folk medicine for the treatment or prevention of diseases. In this context, the purpose of this work was to study the antibacterial activity of crude hydroalcoholic extract from Baccharis trimera against Gram-positive bacterial strains (Staphylococcus aureus ATCC 29213, Staphylococcus saprophyticus ATCC 15305, Staphylococcus epidermidis ATCC 12228, Enterococcus faecalis ATCC 19433) and Gram-negative bacteria (Escherichia coli EHEC ATCC 43895, Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumoniae ATCC 27736, Salmonella typhi ATCC 19430) of clinical interest. Antibacterial susceptibility was evaluated by broth microdilution assay following the CLSI (formerly the NCCLS) guidelines. The extract from B. trimera showed antibacterial activity against Gram-positive bacteria and the most interesting result was obtained against S. epidermidis that presented Minimal Inhibitory Concentration of 250μg/mL. These results indicate that B. trimera have bacterisostatic potential against Gram-positive bacterial strains of medical interest and could serve as a base for further studies on the use of isolated compounds from this species as future antimicrobials.

Growth and yield of corn hybrids in response to association with Azospirillum brasilense and nitrogen fertilization

There is a growing interest in optimizing the positive effects of the association between Azospirillum bacteria and corn crop in order to reduce the use of nitrogen fertilizers. This study aimed to evaluate the inoculation efficiency of an A. brasilense-based commercial product in association with different rates of nitrogen fertilization in two corn genotypes. The experiment was arranged in a 2 x 2 x 5 factorial randomized block design, with four replications. The treatments consisted of two corn hybrids (30F53 and CD386); with and without inoculation with a commercial product based on A. brasilense and five nitrogen rates (0, 40, 80, 120 and 160 kg ha-1). The variables plant height, basal stem diameter, leaf area, shoot dry matter, leaf nitrogen content, length and diameter of the cob, weight of 100 grains and grain yield were evaluated. Inoculation with A. brasilense provided increases of 11 and 12% in leaf area and shoot dry matter, respectively. There were differences in the response of the corn hybrids for most variables and the increase in nitrogen supply provided increments in the growth and yield of corn.

Synthesis, Antimicrobial and Antiproliferative Activity of Novel Silver(I) Tris(pyrazolyl)methanesulfonateand 1,3,5-Triaza-7-phosphadamantane Complexes

Five new silver(I) complexes of formulas [Ag(Tpms)] (1), [Ag(Tpms)-(PPh3)] (2), [Ag(Tpms)(PCy3)] (3), [Ag(PTA)][BF4] (4), and [Ag(Tpms)(PTA)] (5) {Tpms = tris(pyrazol-1-yl)methanesulfonate, PPh3 = triphenylphosphane, PCy3 = tricyclohexylphosphane, PTA = 1,3,5-triaza-7-phosphaadamantane) have been synthesized and fully characterized by elemental analyses, H-1, C-13, and P-31 NMR, electrospray ionization mass spectrometry (ESI-MS), and IR spectroscopic techniques. The single crystal X-ray diffraction study of 3 shows the Tpms ligand acting in the N-3-facially coordinating mode, while in 2 and 5 a N2O-coordination is found, with the SO3 group bonded to silver and a pendant free pyrazolyl ring. Features of the tilting in the coordinated pyrazolyl rings in these cases suggest that this inequivalence is related with the cone angles of the phosphanes. A detailed study of antimycobacterial and antiproliferative properties of all compounds has been carried out. They were screened for their in vitro antimicrobial activities against the standard strains Enterococcus faecalis (ATCC 29922), Staphylococcus aureus (ATCC 25923), Streptococcus pneumoniae (ATCC 49619), Streptococcus pyogenes (SF37), Streptococcus sanguinis (SK36), Streptococcus mutans (UA1S9), Escherichia coli (ATCC 25922), and the fungus Candida albicans (ATCC 24443). Complexes 1-5 have been found to display effective antimicrobial activity against the series of bacteria and fungi, and some of them are potential candidates for antiseptic or disinfectant drugs. Interaction of Ag complexes with deoxyribonucleic acid (DNA) has been studied by fluorescence spectroscopic techniques, using ethidium bromide (EB) as a fluorescence probe of DNA. The decrease in the fluorescence of DNA EB system on addition of Ag complexes shows that the fluorescence quenching of DNA EB complex occurs and compound 3 is particularly active. Complexes 1-5 exhibit pronounced antiproliferative activity against human malignant melanoma (A375) with an activity often higher than that of AgNO3, which has been used as a control, following the same order of activity inhibition on DNA, i.e., 3 > 2 > 1 > 5 > AgNO3 >> 4.

Antimicrobial and cytotoxic assessment of marine cyanobacteria - synechocystis and synechococcus

Aqueous extracts and organic solvent extracts of isolated marine cyanobacteria strains were tested for antimicrobial activity against a fungus, Gram-positive and Gram-negative bacteria and for cytotoxic activity against primary rat hepatocytes and HL-60 cells. Antimicrobial activity was based on the agar diffusion assay. Cytotoxic activity was measured by apoptotic cell death scored by cell surface evaluation and nuclear morphology. A high percentage of apoptotic cells were observed for HL-60 cells when treated with cyanobacterial organic extracts. Slight apoptotic effects were observed in primary rat hepatocytes when exposed to aqueous cyanobacterial extracts. Nine cyanobacteria strains were found to have antibiotic activity against two Gram-positive bacteria, Clavibacter michiganensis subsp. insidiosum and Cellulomonas uda. No inhibitory effects were found against the fungus Candida albicans and Gram-negative bacteria. Marine Synechocystis and Synechococcus extracts induce apoptosis in eukaryotic cells and cause inhibition of Gram-positive bacteria. The different activity in different extracts suggests different compounds with different polarities.

Rhodolith forming coralline algae in the Upper Miocene of Santa Maria Island (Azores, NE Atlantic) : a critical evaluation

The Late Miocene Malbusca outcrop is located in the southeastern coast of Santa Maria Island (Azores, NE Atlantic), interspersed in volcanic formations. At ~20 meters above present sea level, a prominent discontinuous layer of rhodoliths seizes with an extension of ~250 meters. This paper presents the first taxonomic record of fossil rhodolith forming coralline algae for the Miocene of the Azores. The preserved taxonomic features used were the following: (1) arrangement of basal filaments, (2) epithallial cells (when observable), (3) presence of cell fusions, (4) conceptacle type, (5) number of cells layers which conceptacle chamber floors are situated below the surrounding thallus surface and (6) for the sporangial pores, the orientation of the filaments around the conceptacle pores. Based on these characters, six taxa were identified encompassing three Corallinaceae (Lithophyllum prototypum, Lithophyllum sp., Spongites sp., Hydrolithon sp.) and one Hapalidaceae (Phymatolithon calcareum and cf. Phymatolithon sp.). An unidentified coaxial thallus was also present, the coaxial construction ascribing the specimens to the genus Mesophyllum or Neogoniolithon. Taxonomic accounts for the identified taxa are described, illustrated and an identification key is provided. The report of L prototypum represents the first Miocene record and the preservation of the specimens is very good. Miocene coralline algae seem very consistent among deposits but some species are relevant for particular areas, like in the Azores.

Diversity and characterisation of coagulasenegative staphylococci isolated from healthy individuals in Portugal

In the past few years the interest in coagulase-negative staphylococci (CoNS) has significantly increased in human medicine. CoNS are common commensal colonisers of the human skin, although now also recognised as major nosocomial pathogens. Over the last decades, several studies have been carried out in order to understand the pathogenicity mechanisms of CoNS. The well known determinants in the pathogenesis of CoNS infections are their ability to form biofilms and an exceptional resistance to several antibiotics. Nevertheless, there is a lack of studies regarding the commensal lifestyle of these microorganisms. Additionally, it is now hypothesised that commensal bacteria might be a reservoir of pathogenic determinants. Therefore, the work described throughout this thesis was aimed to perform a phenotypic and genotypic characterisation of different CoNS species isolated from healthy Portuguese individuals. A total of 61 CoNS isolates, comprising 7 different species, were obtained and characterised at the level of biofilm formation and antibiotic susceptibility profiles. According to the results, biofilm formation ability and presence of biofilm-associated genes were commonly found features, highlighting their pivotal role in the colonising lifestyle of CoNS. This study also addressed the correlation between phenotypic and genotypic characteristics of biofilm formation, corroborating and raising questions about the importance of some genes in this process. Moreover, it was observed a great proportion of isolates with decreased susceptibility and multiple resistances to some important antibiotics. A significant association between antibiotic resistance and biofilm formation was also demonstrated, and some hypotheses about the nature of such association were provided. Lastly, the expression patterns of two biofilm-associated genes at two distinct biofilm developmental stages were determined, confirming their importance in the accumulative stage of biofilm formation. Overall, the results presented in this thesis indicate that staphylococcal skin flora might be an important reservoir of potentially pathogenic bacteria and, simultaneously, bring to light new perceptions about the molecular basis of staphylococcal biofilm formation, and the nature of the association between antibiotic resistance and biofilm formation.

Characterizing the fungal and bacterial microflora and concentrations in fitness centres

Fitness centres are special places where conditions for microbiological proliferation should be considered. Moisture due to human perspiration and water condensation as a result of human physical activities are prevalent in this type of buildings. Exposure to microbial contaminants is clinically associated with respiratory disorders and people who work out in polluted environments would be susceptible to contaminants. This work studied the indoor air contamination in three gymnasiums in Lisbon. The sampling was performed at two periods: at the opening (morning) and closing (night) of the three gymnasiums. The airborne bacterial and fungal populations were sampled by impaction directly onto Tryptic Soy Agar (for bacteria) and Malt Extract Agar (for fungi) plates, using a Merck MAS-100 air sampler. Higher bacterial concentrations were found at night as compared to the morning but the same behaviour was not found for fungal concentrations. Gram-negative catalase positive cocci were the dominant bacteria in indoor air samples of the studied gymnasiums. In this study, 21 genera/species of fungal colonies were identified. Chrysosporium sp., Chrysonilia sp., Neoscytalidium hialinum, Sepedonium sp. and Penicillium sp. were the most prevalent species identified in the morning, while Cladosporium sp., Penicillium sp., Chrysosporium sp., Acremonium sp. and Chrysonilia sp. were more prevalent at night. A well-designed sanitation and maintenance program for gymnasiums is needed to ensure healthier space for indoor physical activity.

Approaches to tuberculosis mucosal vaccine development using nanoparticles and microparticles: a review

Next-generation vaccines for tuberculosis should be designed to prevent the infection and to achieve sterile eradication of Mycobacterium tuberculosis. Mucosal vaccination is a needle-free vaccine strategy that provides protective immunity against pathogenic bacteria and viruses in both mucosal and systemic compartments, being a promising alternative to current tuberculosis vaccines. Micro and nanoparticles have shown great potential as delivery systems for mucosal vaccines. In this review, the immunological principles underlying mucosal vaccine development will be discussed, and the application of mucosal adjuvants and delivery systems to the enhancement of protective immune responses at mucosal surfaces will be reviewed, in particular those envisioned for oral and nasal routes of administration. An overview of the essential vaccine candidates for tuberculosis in clinical trials will be provided, with special emphasis on the potential different antigens and immunization regimens.

Anatase as an alternative application for preventing biodeterioration of mortars: evaluation and comparison with other biocides

International Biodeterioration & Biodegradation 64(2010)388 e 396

The snakes (Reptilia, Serpentes) of the Miocene of Portugal

The ophidian fauna coming from the Miocene of Amor (MN 5), Quinta das Pedreiras and Quinta do Pombeiro (both MN4) (Portugal) consists of the following taxa: cf. Bavarioboa sp. (Boidae); cf. Coluber sp. (Coluhridae); two unidentified Colubridae; Vipera sp. ("Oriental vipers" complex) (Viperidae). The assemblage is characteristic for the lower/middle Miocene transition and it resembles snake faunas known from other European localities of similar age.

On the hydrogeology of the Lower Tagus Basin and its Cenozoic geologic evolution

The chemical features of the ground water in the Lower Tagus Cenozoic deposits are strongly influenced by lithology, by the velocity and direction of the water movement as well as by the localization of the recharge and discharge zones. The mineralization varies between 80 and 900 mg/l. It is minimal in the recharge zones and in the Pliocene sand and maximum in the Miocene carbonated and along the alluvial valley. Mineralization always reflects the time of permanence, the temperature and the pressure. The natural process of water mineralization is disturbed in agricultural areas because the saline concentration of the infiltration water exceeds that of the infiltrated rainwater. In the discharge zones, the rise of the more mineralized, some times thermal deep waters related to tectonic accidents give rise to anomalies in the distribution of the aquiferous system mineralization model. The diversity of the hydrochemical facies of the ground water may be related to several factors whose identification is some times difficult.

Antibacterial activity of Ionic Liquids based on ampicillin against resistant bacteria

Antibacterial activity of novel Active Pharmaceutical Ingredient Ionic Liquids (API-ILs) based on ampicillin anion [Amp] have been evaluated. They showed growth inhibition and bactericidal properties on some sensitive bacteria and especially some Gram-negative resistant bacteria when compared to the [Na][Amp] and the initial bromide and chloride salts. For these studies were analysed the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBIC) against sensitive Gram-negative bacteria Escherichia coli ATCC 25922 and Klebsiella pneumonia (clinically isolated), as well as sensitive Gram positive S. Aureus ATCC 25923, Staphylococcus epidermidis and Enterococcus faecalis and completed using clinically isolated resistent strains: E. coli TEM CTX M9, E. coli CTX M2 and E. coli AmpC Mox. From the obtained MIC values of studied APIs-ILs and standard [Na][Amp] were derived RDIC values (relative decrease of inhibitory concentration). High RDIC values of [C16Pyr][Amp] especially against two resistant Gram-negative strains E. coli TEM CTX M9 (RDIC>1000) and E. coli CTX M2 (RDIC>100) point clearly to a potential promising role of APIs-ILs as antimicrobial drugs especially against resistant bacterial strains.