Functional taxonomy of bacterial hyperstructures.

The levels of organization that exist in bacteria extend from macromolecules to populations. Evidence that there is also a level of organization intermediate between the macromolecule and the bacterial cell is accumulating. This is the level of hyperstructures. Here, we review a variety of spatially extended structures, complexes, and assemblies that might be termed hyperstructures. These include ribosomal or "nucleolar" hyperstructures; transertion hyperstructures; putative phosphotransferase system and glycolytic hyperstructures; chemosignaling and flagellar hyperstructures; DNA repair hyperstructures; cytoskeletal hyperstructures based on EF-Tu, FtsZ, and MreB; and cell cycle hyperstructures responsible for DNA replication, sequestration of newly replicated origins, segregation, compaction, and division. We propose principles for classifying these hyperstructures and finally illustrate how thinking in terms of hyperstructures may lead to a different vision of the bacterial cell.     

Control of blood-transmitted infections in dentistry

As knowledge has accumulated on the blood-transmitted pathogenic agents, the contact with biological fluids (blood, plasma, saliva, etc.) from apparently healthy individuals has started to be regarded as a real professional risk for dentists. Theoretically, exposure to a contaminated biological specimen may have as a consequence transmission of infection from patient to dentist, from dentist to patient and from patient to patient via inadequately decontaminated and sterilized dental equipment. The present study is concerned with the analysis of the specific conditions that favor the occurrence of the epidemic process, the estimation of the risk degree of transmission of infections caused by hepatitis B, C viruses as well as of HIV infection in Romania. The data for the study were collected using two processes. First a self reporting survey and secondly an experimental procedure were performed. The testing of dentists' knowledge of blood transmissible diseases and infection control in their offices were performed using a questionnaire with 129 questions. The professional incidents/accidents representing a potential risk were counted using a questionnaire (with 37 questions). Serological markers were tested with ELISA kits. The monitoring of sterilization was accomplished with a questionnaire and biological tests. Many conclusions result from the study. There is an extremely reduced probability and infection transmission from the dentist to the patient. The transmission of infection from the patient to the dentist represents a low risk (for all that, the risk should not be minimized). The rigorous control and observation of infection prevention measures in dental offices is necessary to stop the infection transmission from patient to patient. The dentists' postgraduate training in infection control measures should be completed with knowledge regarding the blood transmissible infections epidemiology. Learning more about the epidemiological process enables the dentists to avoid wrong attitudes and behaviors.     

Ecosystem engineers activate mycorrhizal mutualism in salt marshes

Theory predicts that ecosystem engineers should have their most dramatic effects when they enable species, through habitat amelioration, to live in zones where physical and biological conditions would otherwise suppress or limit them. Mutualisms between mycorrhizal fungi and plants are key determinants of productivity and biodiversity in most terrestrial systems, but are thought to be unimportant in wetlands because anoxic sediments exclude fungal symbionts. Our field surveys revealed arbuscular mycorrhizal associations on salt marsh plant roots, but only in the presence of crabs that oxygenate soils as a by-product of burrowing. Field experiments demonstrate that fungal colonization is dependent on crab burrowing and responsible for nearly 35% of plant growth. These results highlight ecosystem engineers as ecological linchpins that can activate and maintain key mutualisms between species. Our findings align salt marshes with other important biogenic habitats whose productivity is reliant on mutualisms between the primary foundation species and micro-organisms.     

Behaviour of bacterial division protein FtsZ under a monolayer with phospholipid domains

Assembly of the tubulin-like protein FtsZ at or near the cytoplasmic membrane is one of the earliest steps in division of bacteria such as Escherichia coli. Exactly what constitutes the site at which FtsZ acts is less clear. To investigate the influence of the membrane phospholipids on FtsZ localization and assembly, we have elaborated with the Langmuir technique a two-lipid monolayer made of dilauryl-phosphatidylethanolamine (DLPE) and dipalmitoyl-phosphatidylglycerol (DPPG). This monolayer comprised stable condensed domains in an expanded continuous phase. In the presence of GTP, FtsZ assembly disrupts the condensed domains within 5 min. After several hours, with or without GTP, FtsZ assembled into large aggregates at the domain interface. We suggest that the GTP-induced polymerization of FtsZ is coupled to the association of FtsZ protofilaments with domain interfaces.     

Behaviour of bacterial division protein FtsZ under a monolayer with phospholipid domains

Assembly of the tubulin-like protein FtsZ at or near the cytoplasmic membrane is one of the earliest steps in division of bacteria such as Escherichia coli. Exactly what constitutes the site at which FtsZ acts is less clear. To investigate the influence of the membrane phospholipids on FtsZ localization and assembly, we have elaborated with the Langmuir technique a two-lipid monolayer made of dilauryl-phosphatidylethanolamine (DLPE) and dipalmitoyl-phosphatidylglycerol (DPPG). This monolayer comprised stable condensed domains in an expanded continuous phase. In the presence of GTP, FtsZ assembly disrupts the condensed domains within 5 min. After several hours, with or without GTP, FtsZ assembled into large aggregates at the domain interface. We suggest that the GTP-induced polymerization of FtsZ is coupled to the association of FtsZ protofilaments with domain interfaces.     

Molecular characterization of the Entamoeba histolytica enolase gene and modelling of the predicted protein

Entamoeba histolytica obtains its energy mainly from glucose fermentation. Enzymes involved in this pathway could be potential targets for antiparasite drugs. Here we report the molecular characterization of the E. histolytica enolase gene (Ehenl-1), which in a single copy is located on the 1.6 Mb chromosome. It is transcribed into a 1.4 kb mRNA which starts 13 nucleotides upstream of the ATG start codon. The sequence TATAAG, at −31, interacted with nuclear proteins suggesting that it has a TATA box function. Protein modelling allowed us to identify a putative specific region that differs from human enolase and could be a good target for the design of novel drugs against E. histolytica.     

Individual variation in feeding morphology,not diet,can facilitate the success of generalist species in urban ecosystems

Generalist species dominate urban ecosystems. The success of urban generalists is often related to a plastic diet and feeding traits that allow them to take advantage of a variety of food resources provided by humans in cities. The classification of a species as a generalist is commonly based on mean estimates of diet‐ and feeding‐related traits. However, there is increasing evidence that a generalist population can consist of individual specialists. In such cases, estimates based on mean can hide important individual variation that can explain trophic ecology and the success of urban dwellers. Here, we focus on guppies, Poecilia reticulata, a widespread alien fish species which has invaded both urban and non‐urban systems, to explore the effect of urbanization on individual diet and feeding morphology (cranium shape). Our results show that guppies in urban and non‐urban populations are not individual specialists, having a similar generalist diet despite the high population density. However, there is important individual variation in cranium shape which allow urban guppies to feed more efficiently on highly nutritious food. Our data suggest that individual variation in feeding efficiency can be a critical overlooked trait that facilitates the success of urban generalists.     

Effectiveness of near-infrared spectroscopy as a non-invasive tool to discriminate spectral profiles of in vitro cultured oocytes from goats

Here, we aimed to discriminate between the spectral profiles of spent culture media after oocyte in vitro maturation (IVM) and culture (IVC) from goats of different ages subjected to repeated hormonal treatments. The profiles were discriminated using near infrared (NIR) spectroscopy combined with multivariate methods. A total of 19 goats (young = 10; old = 9) were subjected to serial hormonal stimulation (HS) with gonadotropins. Cumulus oophorus complexes (COCs) were collected using laparoscopic ovum pick-up (LOPU) and subjected to IVM and parthenogenetic activation. The initial embryos were subjected to IVC. Spent culture media were collected after oocyte IVM and on day 2 of IVC and analyzed using NIR spectroscopy. NIR spectral data were interpreted through chemometric methods, such as principle component analysis (PCA) and partial least square discriminant analysis (PLS-DA). The results of PCA analysis clearly showed a separation in the spectral profiles between the experimental groups (HS sessions; young and old animals) both after IVM and IVC. Overall, the main absorption bands were attributed to the C-H group second overtone, first overtone of O-H and N-H, and C-H combinations and may serve as molecular markers. On the other hand, the spectral data obtained using PLS-DA models provided a better classification of the groups. The results showed the possibility of discriminating young and old groups as well as the three HS sessions with high specificity, sensitivity, and accuracy using NIR spectra. Thus, the culture medium analysis using NIR spectroscopy combined with multivariate methods indicated the dissimilarities between the groups and provided an insight into the in vitro development of goat oocytes. This technique serves as an efficient, objective, rapid, and non-invasive method to discriminate spectral profiles.     

Characterization of senescence- and apoptosis-dependent forms of terminin as derived from a precursor found in replicating and nonreplicating cells

Previously we have reported the production of a monoclonal antibody (Mab 1.2) which recognizes a cytoplasmic protein, terminin, in three different molecular weights: 90 (Tp90), 60 (Tp60), and 30 kDa (Tp30) forms. Further characterization shows that Tp90 is found in young growing and nongrowing quiescent fibroblasts, while Tp60 is found in permanently growth-arrested senescent fibroblasts and Tp30 in cells committed to undergo programmed cell death (apoptosis). In tissue, Tp90 is found in embryonic brain; later, in neonatal brain after terminal differentiation is completed, only Tp60 is found. Tp30 is found in crude liver fractions extracted without the protective action of protease inhibitors. In all these circumstances, Tp90 is mostly seen in the detergent-soluble fraction, while Tp60 and Tp30 are detergent-insoluble. We now report that in cultured fibroblasts, as well as in tissues such as brain and liver, Tp60 and Tp30 are derived from the Tp90 polypeptide, indicated by the fact that only the Tp90 species is identified by both immunoblotting and immunoprecipitation assays, when the cell or tissue extracts are prepared in the presence of protease inhibitors. Further evidence shows that immunoprecipitation of in vitro translation products from brain, liver, and cultured fibroblasts also present a single band of Tp90 polypeptide. Pulse-chase experiments show that during apoptosis, Tp90 is processed to Tp60, and eventually to Tp30. However, when the total protein extracts are fractionated, only Tp90 is found in the detergent-soluble fraction, with diminishing quantities during the time course of apoptosis, and Tp30, in contrast, is found as the only protein species in the insoluble fraction, with increasing quantity during the same time course. Newly processed Tp60 is not found in either of the fractions, reflecting its loss during the fractionation procedure. Limited one-dimensional peptide mapping of Tp90 yields three different bands at 30, 28, and 25 kDa, but only the one at 30 kDa is recognized by Mab 1.2. These results lead us to suggest that terminin protein is synthesized in the Tp90 form, and cleaved to lower molecular weight forms depends upon different physiologic conditions, with Tp60 processed in the terminally differentiated or senescent state and rapidly to Tp30 in apoptosis. Our findings further suggest that Tp90's processing to either Tp60 or Tp30 produces insoluble protein forms. Furthermore, the presence of Tp90 in nonapoptotic (either replicating or nonreplicating) cells may reflect the absence of necessary proteolytic action required for the execution of apoptosis. Future experiments will allow us to determine the nature of this proteolytic action, as well as whether this action is due to the autocatalytic action of Tp90 or by other endogenous proteases, and then to determine the significance of this biochemical action in cells. © 1996 Wiley-Liss, Inc.