Decreased neoblast progeny and increased cell death during starvation-induced planarian degrowth

The development of a complex multicellular organism requires a careful coordination of growth, cell division, cell differentiation and cell death. All these processes must be under intricate and coordinated control, as they have to be integrated across all tissues. Freshwater planarians are especially plastic, in that they constantly replace somatic tissues from a pool of adult somatic stem cells and continuously undergo growth and degrowth as adult animals in response to nutrient availability. During these processes they appear to maintain perfect scale of tissues and organs. These life history traits make them an ideal model system to study growth and degrowth. We have studied the unique planarian process of degrowth. When food is not available, planarians are able to degrow to a minimum size, without any signs of adverse physiological outcomes. For example they maintain full regenerative capacity. Our current knowledge of how this is regulated at the molecular and cellular level is very limited. Planarian degrowth has been reported to result from a decrease in cell number rather than a decrease in cell size. Thus one obvious explanation for degrowth would be a decrease in stem cell proliferation. However evidence in the literature suggests this is not the case. We show that planarians maintain normal basal mitotic rates during degrowth but that the number of stem cell progeny decreases during starvation and degrowth. These observations are reversed upon feeding, indicating that they are dependent on nutritional status. An increase in cell death is also observed during degrowth, which is not rapidly reversed upon feeding. We conclude that degrowth is a result of cell death decreasing cell numbers and that the dynamics of neoblast self-renewal and differentiation adapt to nutrient conditions to allow maintenance of the neoblast population during the period of starvation.     

High-yield secretion of multiple client proteins in Aspergillus

Production of pure and high-yield client proteins is an important technology that addresses the need for industrial applications of enzymes as well as scientific experiments in protein chemistry and crystallization. Fungi are utilized in industrial protein production because of their ability to secrete large quantities of proteins. In this study, we engineered a high-expression-secretion vector, pEXPYR that directs proteins towards the extracellular medium in two Aspergillii host strains, examine the effect of maltose-induced over-expression and protein secretion as well as time and pH-dependent protein stability in the medium. We describe five client proteins representing a core set of hemicellulose degrading enzymes that accumulated up to 50-100 mg/L of protein. Using a recyclable genetic marker that allows serial insertion of multiple genes, simultaneous hyper-secretion of three client proteins in a single host strain was accomplished.     

First description of the male and subgravid female of Philometra serranellicabrillae Janiszewska, 1949 (Nematoda: Philometridae), a gonad-infecting parasite of the comber Serranus cabrilla (Linnaeus) (Serranidae) off Tunisia

Systematic Parasitology - The male and subgravid female of Philometra serranellicabrillae Janiszewska, 1949 (Philometridae) collected from the gonads of Serranus cabrilla (Linnaeus) (Serranidae)...     

A phylogenetic study of the Thygater‐Trichocerapis group and new scopes for the subgenera of Thygater Holmberg, (Hymenoptera,Apidae)

The bee tribe Eucerini is a large monophyletic taxon occurring in all continents, except Oceania and Antarctica, but far more diverse in the Americas and, especially, in the Neotropics. The phylogenetic relationships within its subtribe Eucerina, which contains the bulk of Eucerini diversity, is poorly understood, this being especially true for the relationships among its Neotropical representatives, which have been poorly represented in all phylogenetic studies, including Eucerini, to date. This leads to a generalized lack of confidence on the monophyly of the groups currently accepted as genera and subgenera. Here, a phylogenetic study based on three molecular markers (COI, 18S, and 28S, totalling about 1700 bp) and 58 morphological characters is presented as a contribution to the understanding of the relationships of the so‐called Thygater‐Trichocerapis group, and especially of the genus Thygater Holmberg, which has not previously been extensively sampled. Representatives of Trichocerapis, including its monotypic subgenus T. (Dithygater), are included for the first time in a phylogenetic study. The main results were: (i) support for the monophyly of the Thygater‐Trichocerapis group; (ii) support for the monophyly of Thygater; (iii) recognition of two main clades in Thygater, each one containing the type species of one of the previously recognized subgenera; and (iv) additional support for the position of Alloscirtetica as sister to all remaining Eucerina. Based on these results a redefinition of the scope of the two subgenera of Thygater is proposed, with changes in the subordination of three of its species, T. (Nectarodiaeta) chaetaspis comb.n. , T. (Nectarodiaeta) paranaensis comb.n. and T. (Thygater) mexicana comb.n.     

A Stochastic Model of Nonenzymatic Nucleic Acid Replication: “Elongators” Sequester Replicators

The origin of nucleic acid template replication is a major unsolved problem in science. A novel stochastic model of nucleic acid chemistry was developed to allow rapid prototyping of chemical experiments designed to discover sufficient conditions for template replication. Experiments using the model brought to attention a robust property of nucleic acid template populations, the tendency for elongation to outcompete replication. Externally imposed denaturation-renaturation cycles did not reverse this tendency. For example, it has been proposed that fast tidal cycling could establish a TCR (tidal chain reaction) analogous to a PCR (polymerase chain reaction) acting on nucleic acid polymers, allowing their self-replication. However, elongating side-reactions that would have been prevented by the polymerase in the PCR still occurred in the simulation of the TCR. The same finding was found with temperature and monomer cycles. We propose that if cycling reactors are to allow template replication, oligonucleotide phenotypes that are capable of favorably altering the flux ratio between replication and elongation, for example, by facilitating sequence-specific cleavage within templates, are necessary; accordingly the minimal replicase ribozyme may have possessed restriction functionality.     

Diversity and distribution of orchid bees (Hymenoptera: Apidae) with a revised checklist of species

The aim of this study was to investigate the diversity and distribution patterns of orchid bees (Euglossina). Cluster and correlation analyses were applied to data extracted from 28 orchid-bee surveys throughout the Neotropical Region. The 28 sampling sites were grouped in three main biogeographic areas that roughly correspond to the Amazonian Basin, the Atlantic Forest and Central America. These three regions, as well as subregions within each of them, correspond approximately to biogeographic components identified through phylogeny-based analyses for other bees and organisms. The Amazonian Forest as a whole has the richest fauna and the highest levels of endemism. The Atlantic Forest, on the other hand, showed the poorest fauna and the lowest levels of endemism. However, a major neotropical biome, in which orchid bees are known to occur, has not been sampled yet, the savanna-like cerrado. At least 30% of the species are endemic to each biome. An updated checklist of the species of Euglossina is provided.     

In vivo site-specific recombination using the beta-rec/six system

The prokaryotic beta serine recombinase (beta-rec) catalyzes site-specific recombination between two directly oriented six sites (93 bp) in mammalian cells, both in episomal and in chromosomally integrated substrates. The beta-rec/six exclusive intramolecular site-specific recombination (SSR) system has been proposed as a suitable approach when several independently controlled recombination events are needed in a single cell. Here we explored the use of the beta-rec/six system for selective induction of genome-targeted modifications. We generated and analyzed mouse transgenic lines (Tgbeta) expressing beta-rec under the control of the Lck promoter. beta-rec activity was demonstrated, and there was no evidence of alterations to thymic or peripheral T cell development. We developed two transgenic mouse lines harboring different target sequences (Tgrec and KOsix) and analyzed the effect of beta-rec expression on these animals. The results indicate that the beta-rec/six SSR system is functional for in vivo gene-targeting applications.     

First homo-peptides undergoing a reversible 3(10)-helix/alpha-helix transition: critical main-chain length

The difference in length between the more elongated peptide 3(10)-helix and the more compact alpha-helix is about 0.4 A/residue. This property makes the 3(10)-/alpha-helix reversible conversion very promising as a molecular switching tool between the N- and C-terminal functions of a peptide backbone. In this work, using homo-peptides of various main-chain length, all based on the strongly helicogenic, Calpha-tetrasubstituted alpha-amino acid Calpha-methyl-L-valine, we show that a well defined, solvent controlled, reversible 3(10)-/alpha-helix transition takes place even in a homo-oligomer as short as a terminally blocked hexapeptide. Homo-peptide sequences blocked as a urethane or an acetamide at the N-terminus and as a methyl ester or an N-alkyl amide at the C-terminus are all appropriate. The nature of the occurring helical species in the various solvents tested was assessed by electronic or vibrational circular dichroism.