Insect-Symbiont Gene Expression in the Midgut Bacteriocytes of a Blood-Sucking Parasite.

Animals interact with a diverse array of both beneficial and detrimental microorganisms. In insects, these symbioses in many cases allow feeding on nutritionally unbalanced diets. It is, however, still not clear how are obligate symbioses maintained at the cellular level for up to several hundred million years. Exact mechanisms driving host-symbiont interactions are only understood for a handful of model species and data on blood-feeding hosts with intracellular bacteria are particularly scarce. Here, we analyzed interactions between an obligately blood-sucking parasite of sheep, the louse fly Melophagus ovinus, and its obligate endosymbiont, Arsenophonus melophagi. We assembled a reference transcriptome for the insect host and used dual RNA-Seq with five biological replicates to compare expression in the midgut cells specialized for housing symbiotic bacteria (bacteriocytes) to the rest of the gut (foregut-hindgut). We found strong evidence for the importance of zinc in the system likely caused by symbionts using zinc-dependent proteases when acquiring amino acids, and for different immunity mechanisms controlling the symbionts than in closely related tsetse flies. Our results show that cellular and nutritional interactions between this blood-sucking insect and its symbionts are less intimate than what was previously found in most plant-sap sucking insects. This finding is likely interconnected to several features observed in symbionts in blood-sucking arthropods, particularly their midgut intracellular localization, intracytoplasmic presence, less severe genome reduction, and relatively recent associations caused by frequent evolutionary losses and replacements

Quantitative optische Spektroskopie einzelner metallischer Nanostrukturen

Zur genauen optischen Charakterisierung einzelner Nanostrukturen wurde im Rahmen dieser Arbeit eine neuartige Fernfeld-Spektroskopiemethode entwickelt: das modulierte Gleichweg-Interferometer. Mit ihm ist es möglich, das Streu-, das Absorptions- und das Extinktionsquerschnittsspektrum einer einzelnen Nanostruktur direkt und quantitativ zu bestimmen. Mit dieser Technik untersuchten wir verschiedene Typen metallischer Nanoantennen und verglichen unsere Ergebnisse mit denen der Elektronen-Energieverlust-Spektroskopie sowie mit theoretischen Rechnungen

Polarization-Current-Based FDTD Near-to-Far-Field Transformation

A new near-to-far-field transformation algorithm for three-dimensional finite-different time-domain is presented in this article. This new approach is based directly on the polarization current of the scatterer, not the scattered near fields. It therefore eliminates the numerical errors originating from the spatial offset of the E and H fields, inherent in the standard near-to-far-field transformation. The proposed method is validated via direct comparisons with the analytical Lorentz-Mie solutions of plane waves scattered by large dielectric and metallic spheres with strong forward-scattering lobes.Comment: 8 pages, 2 figures. Submitted to publis

Phylogeny of Amphidinium (Dinophyceae) from Guam and Okinawa, with descriptions of A. pagoense sp. nov. and A. uduigamense sp. nov.

Marine benthic dinoflagellates within the genus Amphidinium were isolated from Guam and Okinawa. Isolated strains were identified to species-level using phylogenetic analyses of 28S rRNA and ITS-5.8S rRNA genes as well as microscopy. Of the six isolated strains, two were new species: A. pagoense sp. nov. and A. uduigamense sp. nov. Other isolates included strains of A. massartii and A. operculatum from Guam, and two strains of A. operculatum from Okinawa. Both new species were described using light and electron microscopy (SEM and TEM). The combination of characteristics that make A. pagoense sp. nov. unique includes a pair of centrally-located pyrenoids, variable cell shape, absence of scales and a long, curved ventral ridge. For A. uduigamense sp. nov., a combination of several morphological features distinguishes it from other species. These include a constriction near the anterior of the hypocone, two centrally located pyrenoids, a longitudinal flagellum inserted in the posterior one-third of the cell, cell size, cell division in the motile stage and the absence of scales. Toxicity was confirmed in these two novel species by testing methanol extracts in an Artemia bioassay. Previously unrecorded ITS rRNA gene sequences from A. operculatum were also sequenced from both locations. Species identified and newly described in this study expand the taxonomic knowledge of Amphidinium in the Pacific.journal articl

Genetic Engineering of an Industrial Strain of Saccharomyces cerevisiae for L-Malic Acid Degradation via an Efficient Malo-Ethanolic Pathway

The optimal ratio of L-malic and L-tartaric acid in relation to other wine components is one of the most important aspects that ultimately determine wine quality during winemaking. Winemakers routinely rely on the judicious use of malolactic fermentation (MLF) after alcoholic fermentation to deacidify and stabilise their wines. However, due to theunreliability of the process and unsuitable sensory modifications in some grape cultivars, especially for fruity-floral wines, MLF is often regarded as problematic and undesirable. Alternative methods for reducing the amounts of L-malic acid in wine will contribute to improving the production of quality wines in the future, especially in coolclimate regions. Most wine yeast strains of Saccharomyces are unable to effectively degrade L-malic acid, whereas the fission yeast Schizosaccharomyces pombe efficiently degrades high concentrations of L-malic acid by means of malo-ethanolic fermentation. However, strains of S. pombe are not suitable for vinification due to the production of undesirable off-flavours. Previously, the 5. pombe malate permease (mael) and malic enzyme (mae2) genes were  successfully expressed under the 3-phosphoglycerate kinase (PGK1) regulatory elements in 5. cerevisiae, resulting in a recombinant laboratory strain of S. cerevisiae with an efficient malo-ethanolic pathway. Stable integration of the S. pombe malo-ethanolic pathway genes has now been obtained through the construction of a unique integration strategy in a commercial wine yeast strain. Co-transformation of the linear integration cassette containing the mael and mae2 genes and PGK1 regulatory elements and a multi-copy plasmid containing the phleomycin-resistance marker into a commercial Saccharomyces cerevisiae strain resulted in the successful transformation and integration of the malo-ethanolic genes. The recombinant 5. cerevisiae strain was successfully cured of phleomycin-resistance plasmid DNA in order to obtain malo-ethanolic yeast containing only yeast-derived DNA. The integrated malo-ethanolic genes were stable in 5. cerevisiae and during synthetic and grape must fermentation, L-malic acid was completely fermented to ethanol without any negative effect on fermentation kinetics and wine quality

Implementasi Branding Produk UMKM Desa Tumpakrejo Guna Meningkatkan Nilai Jual Produk

Pandemi Covid-19 menjadi sebuah krisis baru bagi setiap negara dan berdampak pada sektor ekonomi pada seluruh lapisan masyarakat. Salah satu daerah terdampak pada sektor ekonomi khususnya perdagangan yaitu Desa Tumpakrejo, Kecamatan Gedangan, Kabupaten Malang. Sumber daya alam yang melimpah menjadikan sebagian warga di Desa Tumpakrejo bermata pencaharian sebagai pelaku usaha industry rumah tangga atau UMKM. Bagaimanapun, penerapan kebijakan pembatasan sosial masyarakat menyebabkan turunnya tingkat perekonomian di Desa Tumpakrejo akibat dari aktivitas jual beli antara konsumen dan pedagang menurun. Kegiatan ini bertujuan untuk meningkatkan produksi dan perekonomian masyarakat lokal dengan melakukan kegiatan branding produk-produk industri rumah tangga dan UMKM. Secara rinci, program kegiatan ini meliputi analisis situasi, branding dengan pembuatan banner dan logo berupa stiker produk, pemberian edukasi seputar penjualan dan daya saing produksi serta membantu memasarkan produk melalui platform media sosial. Hasil pelaksanaan kegiatan ini mampu mendukung produsen dalam meningkatkan nilai jual dan memiliki identitas produk serta promosi yang lebih kuat untuk bersaing dengan produk lainnya di luar Desa Tumpakrejo. Respon produsen dan konsumen terkait hasil branding produk sangat baik dan produsen akan menggunakan sticker brand yang telah didesain untuk diaplikasikan terhadap produknya. Kata kunci— Pembangunan ekonomi lokal, Branding produk, Strategi pemasaran, Sektor perekonomian desa    Abstract The Covid-19 pandemic has raised a new crisis for every country and it has an impact on the economic sector at all levels of society. One of the areas affected by the economic sector, especially trade, is Tumpakrejo Village, Gedangan District, Malang Regency, Indonesia. Abundant natural resources make some residents in Tumpakrejo Village a livelihood as home industry business actors or SMEs. However, the implementation of the community social restriction policy caused a decrease in the level of the economy in Tumpakrejo Village as a result of declining buying and selling activities between consumers and traders. This community service aims to increase the production and economy of the local community by carrying out branding activities for home industry products and SMEs. In detail, this activity program includes situation analysis, branding by making banners and logos in the form of product stickers, providing education about sales and production competitiveness and helping to market products through social media platforms. The results of the implementation of this activity can support producers in increasing selling value and have a stronger product identity and promotion to compete with other products outside Tumpakrejo Village. The response of producers and consumers regarding the results of product branding is remarkable and producers will involve brand stickers that have been designed to be applied to their products. Keywords— Local economic development, Product branding, Marketing strategy, Rural economic secto

Sex‐specific expression and DNA methylation in a species with extreme sexual dimorphism and paternal genome elimination

Phenotypic differences between sexes are often mediated by differential expression and alternative splicing of genes. However, the mechanisms that regulate these expression and splicing patterns remain poorly understood. The mealybug, Planococcus citri, displays extreme sexual dimorphism and exhibits an unusual instance of sex-specific genomic imprinting, paternal genome elimination (PGE), in which the paternal chromosomes in males are highly condensed and eliminated from the sperm. Planococcus citri has no sex chromosomes and both sexual dimorphism and PGE are predicted to be under epigenetic control. We recently showed that P. citri females display a highly unusual DNA methylation profile for an insect species, with the presence of promoter methylation associated with lower levels of gene expression. Here, we therefore decided to explore genome-wide differences in DNA methylation between male and female P. citri using whole-genome bisulphite sequencing. We identified extreme differences in genome-wide levels and patterns between the sexes. Males display overall higher levels of DNA methylation which manifest as more uniform low levels across the genome. Whereas females display more targeted high levels of methylation. We suggest these unique sex-specific differences are due to chromosomal differences caused by PGE and may be linked to possible ploidy compensation. Using RNA-Seq, we identify extensive sex-specific gene expression and alternative splicing, but we find no correlation with cis-acting DNA methylation

Human airway tuft cells influence the mucociliary clearance through cholinergic signalling

Background Airway tuft cells, formerly called brush cells have long been described only morphologically in human airways. More recent RNAseq studies described a chemosensory cell population, which includes tuft cells, by a distinct gene transcription signature. Yet, until which level in the tracheobronchial tree in native human airway epithelium tuft cells occur and if they function as regulators of innate immunity, e.g., by regulating mucociliary clearance, remained largely elusive. Methods We performed immunohistochemistry, RT-PCR and immunoblotting analyses for various tuft cell markers to confirm the presence of this cell type in human tracheal samples. Immunohistochemistry was conducted to study the distribution of tuft cells along the intrapulmonary airways in humans. We assessed the influence of bitter substances and the taste transduction pathway on mucociliary clearance in mouse and human tracheal samples by measuring particle transport speed. Results Tuft cells identified by the expression of their well-established marker POU class 2 homeobox 3 (POU2F3) were present from the trachea to the bronchioles. We identified choline acetyltransferase in POU2F3 expressing cells as well as the transient receptor potential melastatin 5 (TRPM5) channel in a small population of tracheal epithelial cells with morphological appearance of tuft cells. Application of bitter substances, such as denatonium, led to an increase in mucociliary clearance in human tracheal preparations. This was dependent on activation of the TRPM5 channel and involved cholinergic and nitric oxide signalling, indicating a functional role for human tuft cells in the regulation of mucociliary clearance. Conclusions We were able to detect tuft cells in the tracheobronchial tree down to the level of the bronchioles. Moreover, taste transduction and cholinergic signalling occur in the same cells and regulate mucociliary clearance. Thus, tuft cells are potentially involved in the regulation of innate immunity in human airways

Impact of targeted counseling on reported vaginal hygiene practices and bacterial vaginosis: the HIV Prevention Trials Network 035 study

We describe the impact of intense counseling to reduce vaginal hygiene practices and its effect on bacterial vaginosis (BV)