Evolution of the integral membrane desaturase gene family in moths and flies

Lepidopteran insects use sex pheromones derived from fatty acids in their species-specific mate recognition system. Desaturases play a particularly prominent role in the generation of structural diversity in lepidopteran pheromone biosynthesis as a result of the diverse enzymatic properties they have evolved. These enzymes are homologous to the integral membrane desaturases, which play a primary role in cold adaptation in eukaryotic cells. In this investigation, we screened for desaturase-encoding sequences in pheromone glands of adult females of eight lepidopteran species. We found, on average, six unique desaturase-encoding sequences in moth pheromone glands, the same number as is found in the genome database of the fly, Drosophila melanogaster, vs. only one to three in other characterized eukaryotic genomes. The latter observation suggests the expansion of this gene family in insects before the divergence of lepidopteran and dipteran lineages. We present the inferred homology relationships among these sequences, analyze nonsynonymous and synonymous substitution rates for evidence of positive selection, identify sequence and structural correlates of three lineages containing characterized enzymatically distinct desaturases, and discuss the evolution of this sequence family in insects.     

Butyrate regulation of glycosylation-related gene expression: evidence for galectin-1 upregulation in human intestinal epithelial goblet cells

Glycosylation of mucins produced by human intestinal goblet cells plays a crucial role in their functions: mucus gel physico-chemical protective properties, host-bacteria interactions, cell-cell adhesion, cell migration, and cell signaling. Colonic mucin glycosylation can be modified by luminal metabolites of fiber fermentation like butyrate. Our aim was to assess the effect of butyrate on the expression of a large panel of glycosylation-related genes in human intestinal epithelial goblet cells HT29-Cl.16E. We found that only a very scarce group of genes: 9 out of 252 were evidenced by microarray screening, and only three had their modulation significantly confirmed by real time PCR quantification. The most striking effect of butyrate was its 8- to 18-fold increase of galectin-1 gene expression, which was confirmed at the protein level, specifically with a central and apical intracellular localization. Significant butyrate effects will be discussed in regard to their possible link with mucins expressed by HT29-Cl.16E cells.     

Antibodies to the CFTR modulate the turgor pressure of guard cell protoplasts via slow anion channels

The plasma membrane guard cell slow anion channel is a key element at the basis of water loss control in plants allowing prolonged osmolite efflux necessary for stomatal closure. This channel has been extensively studied by electrophysiological approaches but its molecular identification is still lacking. Recently, we described that this channel was sharing some similarities with the mammalian ATP-binding cassette protein, cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel [Leonhardt, N. et al. (1999) Plant Cell 11, 1141-1151]. Here, using the patch-clamp technique and a bioassay, consisting in the observation of the change in guard cell protoplasts volume, we demonstrated that a functional antibody raised against the mammalian CFTR prevented ABA-induced guard cell protoplasts shrinking and partially inhibited the slow anion current. Moreover, this antibody immunoprecipitated a polypeptide from guard cell protein extracts and immunolabeled stomata in Vicia faba leaf sections. These results indicate that the guard cell slow anion channel is, or is closely controlled by a polypeptide, exhibiting one epitope shared with the mammalian CFTR.     

The Ban Don Mun artifacts: A chronological reappraisal of human occupations in the Lampang province of Northern Thailand

Despite recent stone tool evidence demonstrating a much older Early Pleistocene human presence in India, the timing and geography of human demographic expansions in continental Southeast Asia remains ambiguous. The recent discovery of a series of stone artifacts spread over a basalt level at Ban Don Mun in the Lampang province of northern Thailand presents an ideal opportunity for reevaluating lithic assemblages documented during the 1970s and 1980s in the same region. Both the position of these stone tools and new absolute dates indicate a Middle Pleistocene age and call into question the status of these artifacts as the oldest yet found in Southeast Asia. The uncertain geo-chronological context and technological analysis of the chopper industry from previous work in the Lampang area prompted us to undertake new surveys in continental Southeast Asia in order to help clarify the route and timing of Pleistocene human expansions in this part of the world.     

Ethanol Tolerance in the Yeast Saccharomyces cerevisiae Is Dependent on Cellular Oleic Acid Content

In this investigation, we examined the effects of different unsaturated fatty acid compositions of Saccharomyces cerevisiae on the growth-inhibiting effects of ethanol. The unsaturated fatty acid (UFA) composition of S. cerevisiae is relatively simple, consisting almost exclusively of the mono-UFAs palmitoleic acid (Δ⁹Z-C_16:1) and oleic acid (Δ⁹Z-C_18:1), with the former predominating. Both UFAs are formed in S. cerevisiae by the oxygen- and NADH-dependent desaturation of palmitic acid (C_16:0) and stearic acid (C_18:0), respectively, catalyzed by a single integral membrane desaturase encoded by the OLE1 gene. We systematically altered the UFA composition of yeast cells in a uniform genetic background (i) by genetic complementation of a desaturase-deficient ole1 knockout strain with cDNA expression constructs encoding insect desaturases with distinct regioselectivities (i.e., Δ⁹ and Δ¹¹) and substrate chain-length preferences (i.e., C_16:0 and C_18:0); and, (ii) by supplementation of the same strain with synthetic mono-UFAs. Both experimental approaches demonstrated that oleic acid is the most efficacious UFA in overcoming the toxic effects of ethanol in growing yeast cells. Furthermore, the only other UFA tested that conferred a nominal degree of ethanol tolerance is cis-vaccenic acid (Δ¹¹Z-C_18:1), whereas neither Δ¹¹Z-C_16:1 nor palmitoleic acid (Δ⁹Z-C_16:1) conferred any ethanol tolerance. We also showed that the most ethanol-tolerant transformant, which expresses the insect desaturase TniNPVE, produces twice as much oleic acid as palmitoleic acid in the absence of ethanol and undergoes a fourfold increase in the ratio of oleic acid to palmitoleic acid in response to exposure to 5% ethanol. These findings are consistent with the hypothesis that ethanol tolerance in yeast results from incorporation of oleic acid into lipid membranes, effecting a compensatory decrease in membrane fluidity that counteracts the fluidizing effects of ethanol.     

Biofilm dispersal: multiple elaborate strategies for dissemination of bacteria with unique properties

In most environments, microorganisms evolve in a sessile mode of growth, designated as biofilm, which is characterized by cells embedded in a self‐produced extracellular matrix. Although a biofilm is commonly described as a “cozy house” where resident bacteria are protected from aggression, bacteria are able to break their biofilm bonds and escape to colonize new environments. This regulated process is observed in a wide variety of species; it is referred to as biofilm dispersal, and is triggered in response to various environmental and biological signals. The first part of this review reports the main regulatory mechanisms and effectors involved in biofilm dispersal. There is some evidence that dispersal is a necessary step between the persistence of bacteria inside biofilm and their dissemination. In the second part, an overview of the main methods used so far to study the dispersal process and to harvest dispersed bacteria was provided. Then focus was on the properties of the biofilm‐dispersed bacteria and the fundamental role of the dispersal process in pathogen dissemination within a host organism. In light of the current body of knowledge, it was suggested that dispersal acts as a potent means of disseminating bacteria with enhanced colonization properties in the surrounding environment.     

The perforated stones of the Doi Pha Kan burials (Northern Thailand): A Mesolithic singularity?

Throughout continental Southeast Asia, the Hoabinhian techno-complex stands out in clear contrast with the universal chrono-cultural model essentially established on the basis of western prehistory. Following this model, early authors considered perforated stones and associated lithic artefacts as markers of what was then believed to pertain to a Southeast Asian Mesolithic. However, Southeast Asian Mesolitithic has progressively been abandoned in favour of a ubiquitous Hoabinhian spanning from 30,000 to 3000 BP. Here, we present and discuss the discovery of perforated stones at the Doi Pha Kan site in northern Thailand. Perforated stones have almost never been found in undisturbed stratigraphic conditions nor dated with any sufficient degree of certainty. At Doi Pha Kan site, such a kind of artefacts was found in burials intersecting sedimentary layers that could be ascertained as Hoabinhian. In contrast with similar perforated stones described in the literature, that found at Doi Pha Kan are well-dated (13,000 BP), thus providing a time-reference for a putative Southeast Asian Mesolithic. We therefore advocate that such non-Hoabinhian artefacts support the early authors’ hypothesis of the existence of a Southeast Asian Mesolithic. Finally, the funerary practices, the unusually high stature of individuals found at Doi Pha Kan in conjunction with the particular lithic assemblages further contributes to raise the question of the co-occurrence of several cultures or populations at the Pleistocene–Holocene interface in continental Southeast Asia.     

Multi-millennial occupation in northwestern Laos: Preliminary results of excavations at the Ngeubhinh Mouxeu rock-shelter

With over half a century of political instability, resulting from armed conflicts, decolonisation and the Cold War, archaeological investigations in Laos have been rare, leaving little more than a blank page in the chapter of Southeast Asia's prehistory. Recent research has shown that Laos holds a rich prehistoric heritage. In conjunction with the research initiated by J. White who conducted the first professional archaeological survey of northern Laos since decades, we have extended the investigations to the Luang Namtha province. This work allowed us to gather important data about Hoabinhian stone tool assemblages and former cultures. In particular, the archaeological remains and dating from the Ngeubhinh Mouxeu rock-shelter indicate that this mountainous region of Laos has been inhabited over a long period of time that possibly spans as far back as 56,000 ± 3000 BP.