Dissection of the network of interactions that links RNA processing with glycolysis in the Bacillus subtilis degradosome

The RNA degradosome is a multiprotein macromolecular complex that is involved in the degradation of messenger RNA in bacteria. The composition of this complex has been found to display a high degree of evolutionary divergence, which may reflect the adaptation of species to different environments. Recently, a degradosome-like complex identified in Bacillus subtilis was found to be distinct from those found in proteobacteria, the degradosomes of which are assembled around the unstructured C-terminus of ribonuclease E, a protein not present in B. subtilis. In this report, we have investigated in vitro the binary interactions between degradosome components and have characterized interactions between glycolytic enzymes, RNA-degrading enzymes, and those that appear to link these two cellular processes. The crystal structures of the glycolytic enzymes phosphofructokinase and enolase are presented and discussed in relation to their roles in the mediation of complex protein assemblies. Taken together, these data provide valuable insights into the structure and dynamics of the RNA degradosome, a fascinating and complex macromolecular assembly that links RNA degradation with central carbon metabolism.     

Outdoor thermal comfort study in a sub-tropical climate: a longitudinal study based in Hong Kong

This paper presents the findings of an outdoor thermal comfort study conducted in Hong Kong using longitudinal experiments—an alternative approach to conventional transverse surveys. In a longitudinal experiment, the thermal sensations of a relatively small number of subjects over different environmental conditions are followed and evaluated. This allows an exploration of the effects of changing climatic conditions on thermal sensation, and thus can provide information that is not possible to acquire through the conventional transverse survey. The paper addresses the effects of changing wind and solar radiation conditions on thermal sensation. It examines the use of predicted mean vote (PMV) in the outdoor context and illustrates the use of an alternative thermal index—physiological equivalent temperature (PET). The paper supports the conventional assumption that thermal neutrality corresponds to thermal comfort. Finally, predictive formulas for estimating outdoor thermal sensation are presented as functions of air temperature, wind speed, solar radiation intensity and absolute humidity. According to the formulas, for a person in light clothing sitting under shade on a typical summer day in Hong Kong where the air temperature is about 28°C and relative humidity about 80%, a wind speed of about 1.6 m/s is needed to achieve neutral thermal sensation.     

Extracellular and intracellular esterase processing of SCFA–hexosamine analogs: Implications for metabolic glycoengineering and drug delivery

This report provides a synopsis of the esterase processing of short chain fatty acid (SCFA)-derivatized hexosamine analogs used in metabolic glycoengineering by demonstrating that the extracellular hydrolysis of these compounds is comparatively slow (e.g., with a t_1/2 of ～4 h to several days) in normal cell culture as well as in high serum concentrations intended to mimic in vivo conditions. Structure–activity relationship (SAR) analysis of common sugar analogs revealed that O-acetylated and N-azido ManNAc derivatives were more refractory against extracellular inactivation by FBS than their butanoylated counterparts consistent with in silico docking simulations of Ac₄ManNAc and Bu₄ManNAc to human carboxylesterase 1 (hCE1). By contrast, all analogs tested supported increased intracellular sialic acid production within 2 h establishing that esterase processing once the analogs are taken up by cells is not rate limiting.     

The β-subunit of the SnRK1 complex is phosphorylated by the plant cell death suppressor Adi3

The protein kinase AvrPto-dependent Pto-interacting protein3 (Adi3) is a known suppressor of cell death, and loss of its function has been correlated with cell death induction during the tomato (Solanum lycopersicum) resistance response to its pathogen Pseudomonas syringae pv tomato. However, Adi3 downstream interactors that may play a role in cell death regulation have not been identified. We used a yeast two-hybrid screen to identify the plant SnRK1 (for Sucrose non-Fermenting-1-Related Protein Kinase1) protein as an Adi3-interacting protein. SnRK1 functions as a regulator of carbon metabolism and responses to biotic and abiotic stresses. SnRK1 exists in a heterotrimeric complex with a catalytic α-subunit (SnRK1), a substrate-interacting β-subunit, and a regulatory γ-subunit. Here, we show that Adi3 interacts with, but does not phosphorylate, the SnRK1 α-subunit. The ability of Adi3 to phosphorylate the four identified tomato β-subunits was also examined, and it was found that only the Galactose Metabolism83 (Gal83) β-subunit was phosphorylated by Adi3. This phosphorylation site on Gal83 was identified as serine-26 using a mutational approach and mass spectrometry. In vivo expression of Gal83 indicates that it contains multiple phosphorylation sites, one of which is serine-26. An active SnRK1 complex containing Gal83 as the β-subunit and sucrose nonfermenting4 as the γ-subunit was constructed to examine functional aspects of the Adi3 interaction with SnRK1 and Gal83. These assays revealed that Adi3 is capable of suppressing the kinase activity of the SnRK1 complex through Gal83 phosphorylation plus the interaction with SnRK1 and suggested that this function may be related to the cell death suppression activity of Adi3.     

Yeast and yeast-like fungi associated with dry indehiscent fruits of Nothofagus nervosa in Patagonia, Argentina

Nothofagus nervosa (Raulí) is a native tree species that yields valuable timber. It was overexploited in the past and is currently included in domestication and conservation programs. Several research programs have focused on the characterization of epiphytic microorganisms because it has been demonstrated that they can affect plant-pathogen interactions and/or promote plant growth. Although the microbial ecology of leaves has been well studied, less is known about microorganisms occurring on seeds and noncommercial fruits. In this work, we analyzed the yeast and yeast-like fungi present on N.?nervosa fruits destined for the propagation of this species, as well as the effects of fruit preservation and seed dormancy-breaking processes on fungal diversity. Morphological and molecular methods were used, and differences between fungal communities were analyzed using a similarity index. A total of 171 isolates corresponding to 17 species were recovered, most of which belong to the phylum Ascomycota. The majority of the species develop mycelia, produce pigments and mycosporines, and these adaptation strategies are discussed. It was observed that the preservation process considerably reduced yeast and yeast-like fungal diversity. This is the first study concerning microbial communities associated with this ecologically and economically important species, and the information presented is relevant to domestication programs.     

Host cell invasion by Trypanosoma cruzi: a unique strategy that promotes persistence

The intracellular protozoan parasite Trypanosoma cruzi is the causative agent of Chagas' disease, a serious disorder that affects millions of people in Latin America. Despite the development of lifelong immunity following infections, the immune system fails to completely clear the parasites, which persist for decades within host tissues. Cardiomyopathy is one of the most serious clinical manifestations of the disease, and a major cause of sudden death in endemic areas. Despite decades of study, there is still debate about the apparent preferential tropism of the parasites for cardiac muscle, and its role in the pathology of the disease. In this review, we discuss these issues in light of recent observations, which indicate that T. cruzi invades host cells by subverting a highly conserved cellular pathway for the repair of plasma membrane lesions. Plasma membrane injury and repair is particularly prevalent in muscle cells, suggesting that the mechanism used by the parasites for cell invasion may be a primary determinant of tissue tropism, intracellular persistence, and Chagas' disease pathology.     

Mercury methylation and hydrogen sulfide production among unexpected strains isolated from periphyton of two macrophytes of the Amazon

The periphyton of macrophytes had previously been identified as important spots for mercury methylation in the Amazon basin, but the microorganisms that facilitate methylation in such compartment are still to be identified. Here, bacteria were isolated from periphyton associated with Eichhornia crassipes and Polygonum densiflorum in Widdel and Pfennig medium and tested for mercury methylation with a stable isotope tracer technique using (198)HgCl, hydrogen sulfide production and molybdate inhibition. Three Pleomorphomona spp., one unidentified Deltaproteobacteria, two Klebsiella spp., and one Tolumonas sp. were isolated. All except Tolumonas sp. were able to methylate mercury (up to 5% of the (198)HgCl added) and produce up to 4 mM of H(2)S, while the Deltaproteobacteria was also able to demethylate methylmercury. Although these bacteria may not be as strong mercury methylators as sulfate-reducing bacteria, they have the potential to contribute to methylmercury accumulation in the system.     

Characterization of Bifidobacterium spp. strains for the treatment of enteric disorders in newborns

Several studies support the use of probiotics for the treatment of minor gastrointestinal problems in infants. Positive effects on newborn colics have been evidenced after administration of Lactobacillus strains, whereas no studies have been reported regarding the use of bifidobacteria for this purpose. This work was therefore aimed at the characterization of Bifidobacterium strains capable of inhibiting the growth of pathogens typical of the infant gastrointestinal tract and of coliforms isolated from colic newborns. Among the 46 Bifidobacterium strains considered, 16 showed high antimicrobial activity against potential pathogens; these strains were further characterized from a taxonomic point of view, for the presence and transferability of antibiotic resistances, for citotoxic effects and adhesion to nontumorigenic gut epithelium cell lines. Moreover, their ability to stimulate gut health by increasing the metabolic activity and the immune response of epithelial cells was also studied. The examination of all these features allowed to identify three Bifidobacterium breve strains and a Bifidobacterium longum subsp. longum strain as potential probiotics for the treatments of enteric disorders in newborns such as infantile colics. A validation clinical trial involving the selected strains is being planned.     

HPV-16 E2 contributes to induction of HPV-16 late gene expression by inhibiting early polyadenylation

We provide evidence that the human papillomavirus (HPV) E2 protein regulates HPV late gene expression. High levels of E2 caused a read-through at the early polyadenylation signal pAE into the late region of the HPV genome, thereby inducing expression of L1 and L2 mRNAs. This is a conserved property of E2 of both mucosal and cutaneous HPV types. Induction could be reversed by high levels of HPV-16 E1 protein, or by the polyadenylation factor CPSF30. HPV-16 E2 inhibited polyadenylation in vitro by preventing the assembly of the CPSF complex. Both the N-terminal and hinge domains of E2 were required for induction of HPV late gene expression in transfected cells as well as for inhibition of polyadenylation in vitro. Finally, overexpression of HPV-16 E2 induced late gene expression from a full-length genomic clone of HPV-16. We speculate that the accumulation of high levels of E2 during the viral life cycle, not only turns off the expression of the pro-mitotic viral E6 and E7 genes, but also induces the expression of the late HPV genes L1 and L2.     

Myrionema strangulans (Chordariales, Phaeophyceae) epiphyte on Ulva spp. (Ulvophyceae) from Patagonian Atlantic coasts

Fronds of Ulva spp. from Patagonian Atlantic coasts exhibited brown spots produced by the presence of Myrionema strangulans (Chordariales, Phaeophyceae). The occurrence of M. strangulans on Ulva spp. is widely reported from several regions of the world, but there were no detailed studies about the subject. In the present study, we describe the morphology and interactions of M. strangulans with Ulva spp. as observed under light and electron microscopes, and we reconstruct all stages of its life cycle based upon in vitro experiments. The prevalence of infection by M. strangulans was 100%. In case of the strongest epiphytism, the host cuticle exhibited perforations, massive depigmentation, cellular disorganization, and cuticle rupture. It was possible to demonstrate a purely epiphytic life strategy of the organism by transmission electron microscopy. M. strangulans formed discoid thalli constituted by vegetative filaments and radiating from a central zone to a peripheral zone. Transversally, the discs were formed by two strata: a basal monostromatic and a filamentous erect stratum. From the monostromatic stratum, hyaline hairs and reproductive structures were produced. Both plurilocular and unilocular sporangia were present. Zoids from both plurilocular and unilocular sporangia were able to germinate in culture. M. strangulans exhibited a haploid–diploid, heteromorphic life cycle with thalli with three different morphologies. The haploid chromosome number was 12?±?2 chromosomes.