Characteristics of 17Paracoccidioides brasiliensis isolates

We have studied the physiological and morphological features of 17 isolates ofParacoccidioides brasiliensis in order to define their phenotypes. The isolates were cultured at room temperature on potato dextrose agar (PDA, Difco) slants for mycelial growth and in 1% dextrose brain heart infusion agar (BHIA, Difco) at 37°C for the study of yeast forms. Most mycelial and yeast forms grew well between pH 5.6–9.4. In their response to osmotic pressure the isolates were separated in three groups: intolerant, intermediate and tolerant. They also varied in carbohydrate assimilation tests, which indicated important metabolic variation. No clear differences were observed in phenol oxidase tests, KNO₃, starch, casein and arbutin assimilation tests. Only 1 of the isolates, Bt-19, had gelatinase activity. No correlation was observed between the above differences and virulence. Two patterns of growth were observed in the mycelial cultures, glabrous and cottonous, the latter being correlated with increased virulence for ddY mice. Most yeast forms grew as cerebriform colonies, but Pb-HC and Bt-19 colonies had a cobblestone-like surface.     

Pneumococcal Bacteriophage Cp-1 Encodes Its Own Protease Essential for Phage Maturation

The major capsid protein of the pneumococcal phage Cp-1 that accounts for 90% of the total protein found in the purified virions is synthesized by posttranslational processing of the product of the open reading frame (ORF) orf9. Cloning of different ORFs of the Cp-1 genome in Escherichia coli and Streptococcus pneumoniae combined with Western blot analysis of the expressed products led to the conclusion that the product of orf13 is an endoprotease that cleaves off the first 48 amino acid residues of the major head protein. This protease appears to be a key enzyme in the morphopoietic pathway of the Cp-1 phage head. To our knowledge, this is the first case of a bacteriophage infecting gram-positive bacteria that encodes a protease involved in phage maturation.     

The effect of penicillin on competence in Bacillus subtilis cultures growing in chemostat at different doubling times

Summary The relationships between penicillin-resistance and competence at different doubling-times have been studied in a chemostat. Both subinhibitory and lethal levels of drug have a differential effect on competent fraction and cell viability. Transformation frequency was rather increased in the presence of inhibitory doses of penicillin when partially affected cells were protected from lysis by using an isotonic medium; however, transformation frequency decreases when protoplasts were detected.The influence of the doubling-time in the ability of the recipient population to undergo transformation was confirmed. Our results, in accordance with that previously found, might suggest that competence in log phase and competence in the stationary phase could differ in some important aspects.     

Circumtropical distribution of the epiphytic dinoflagellate Coolia malayensis (Dinophyceae): Morphology and molecular phylogeny from Puerto Rico and Brazil

The dinoflagellate genus Coolia, which contains potentially toxic species, is an important component of epiphytic assemblages in marine ecosystems. The morphology of C. malayensis has been illustrated from strains isolated in Asia and Oceania. In this study, strains of C. malayensis isolated from the Caribbean Sea in Puerto Rico, and for the first time from the South Atlantic Ocean in Brazil, were investigated by light, epifluorescence and scanning electron microscopies. No significant morphological differences between these new strains and other geographically distant strains of C. malayensis were observed. In the LSU rDNA phylogeny, the C. malayensis sequences from Brazil and Puerto Rico branched within the clade of strains from Oceania and Asia. The recently described species C. santacroce branched as a sister group of C. monotis, and C. palmyrensis was basal to the combined group of C. monotis/C. malayensis/C. santacroce. A tentative undescribed species from Florida and New Zealand branched as a sister group of C. malayensis. Our results confirm that C. malayensis showed a cosmopolitan distribution in tropical to subtropical waters, while the type species C. monotis remains endemic for the Mediterranean Sea and the temperate North Atlantic.     

Allosteric communication in myosin V: from small conformational changes to large directed movements

The rigor to post-rigor transition in myosin, a consequence of ATP binding, plays an essential role in the Lymn-Taylor functional cycle because it results in the dissociation of the actomyosin complex after the powerstroke. On the basis of the X-ray structures of myosin V, we have developed a new normal mode superposition model for the transition path between the two states. Rigid-body motions of the various subdomains and specific residues at the subdomain interfaces are key elements in the transition. The allosteric communication between the nucleotide binding site and the U50/L50 cleft is shown to result from local changes due to ATP binding, which induce large amplitude motions that are encoded in the structure of the protein. The triggering event is the change in the interaction of switch I and the P-loop, which is stabilized by ATP binding. The motion of switch I, which is a relatively rigid element of the U50 subdomain, leads directly to a partial opening of the U50/L50 cleft; the latter is expected to weaken the binding of myosin to actin. The calculated transition path demonstrates the nature of the subdomain coupling and offers an explanation for the mutual exclusion of ATP and actin binding. The mechanism of the uncoupling of the converter from the motor head, an essential part of the transition, is elucidated. The origin of the partial untwisting of the central beta-sheet in the rigor to post-rigor transition is described.     

Traditional Knowledge and Management of <Emphasis Type="Italic">Feijoa (Acca sellowiana)</Emphasis> in Southern Brazil

Traditional Knowledge and Management of Feijoa (Acca sellowiana) in Southern Brazil. This paper investigates traditional knowledge of the use and management of Acca sellowiana in southern Brazil. Fifty-six informants from three rural communities were assigned to one of four subgroups (“maintainers,” “managers,” “cultivators,” or “users”) based on their responses regarding management and use of A. sellowiana. Traditional knowledge related to use of this species is widespread among rural residents, but traditional knowledge related to management is fragmented depending on whether one uses, manages, or cultivates the species. Knowledge held in rural communities suggests that A. sellowiana could play an expanded role in local economies as well as biodiversity conservation. We suggest that participatory research could stimulate greater local use as well as on-farm conservation of A. sellowiana.     

Presence and environmental circulation of enteric viruses in three different wastewater treatment plants

Aims:  The aim of the work was to evaluate the circulation of the viruses and to determine a correlation between faecal indicators and viruses.
Methods and Results:  Raw wastewater and effluent samples were collected from three wastewater treatment plants, during three sampling periods, and analysed, using cultural and molecular methods, to determine bacteria and virus presence. The results show a removal of bacterial indicators, but a limited reduction of the phages. The viral analysis displays the circulation of cultivable enteroviruses and differences in the seasonal-geographical distribution. Hepatitis A virus was found with only two genotypes: IA-IB. Rotavirus was present in 11·11%, 24·14%, 2·78% of the samples in the 1st, 2nd and 3rd sampling periods; Astrovirus in 33·33%, 6·9%, 25%; Adenovirus in 7·41%, 3·45%, 2·78%; Norovirus in 7·41%, 10·34%, 5·56% respectively. Adenovirus was never identified in plants B and C as Rotavirus in plant C.
Conclusions:  The presence of faecal indicators was not predictive of the enteric virus presence, whereas a different circulation of Enteroviruses was found in the wastewater treatment plants.
Significance and Impact of the Study:  The study shows the importance and the usefulness of molecular methods to evaluate the virus circulation and the genetic variability of Enteroviruses.     

Preparation of fluorescent tocopherols for use in protein binding and localization with the alpha-tocopherol transfer protein

Sixteen fluorescent analogues of the lipid-soluble antioxidant vitamin alpha-tocopherol were prepared incorporating fluorophores at the terminus of omega-functionalized 2-n-alkyl-substituted chromanols (1a-d and 4a-d) that match the methylation pattern of alpha-tocopherol, the most biologically active form of vitamin E. The fluorophores used include 9-anthroyloxy (AO), 7-nitrobenz-2-oxa-1,3-diazole (NBD), N-methyl anthranilamide (NMA), and dansyl (DAN). The compounds were designed to function as fluorescent reporter ligands for protein-binding and lipid transfer assays. The fluorophores were chosen to maximize the fluorescence changes observed upon moving from an aqueous environment (low fluorescence intensity) to an hydrophobic environment such as a protein's binding site (high fluorescence intensity). Compounds 9d (anthroyloxy) and 10d (nitrobenzoxadiazole), having a C9-carbon chain between the chromanol and the fluorophore, were shown to bind specifically and reversibly to recombinant human tocopherol transfer protein (alpha-TTP) with dissociation constants of approximately 280 and 60 nM, respectively, as compared to 25 nM for the natural ligand 2R,4'R,8'R-alpha-tocopherol. Thus, compounds have been prepared that allow the investigation of the rate of alpha-TTP-mediated inter-membrane transfer of alpha-tocopherol and to investigate the mechanism of alpha-TTP function at membranes of different composition.