Miracle-dz/UniFAR-FTUs huggingface dataset:
The Vibrio cholerae Pst2 Phosphate Transport System Is Upregulated in Biofilms and Contributes to Biofilm-Induced Hyperinfectivity

ABSTRACT Vibrio cholerae is the causative agent of the deadly diarrheal disease cholera. As part of its life cycle, V. cholerae persists in marine environments, where it forms surface-attached communities commonly described as biofilms. Evidence indicates that these biofilms constitute the infectious form of the pathogen during outbreaks. Previous work has shown that biofilm-derived V. cholerae cells, even when fully dispersed from the biofilm matrix, are vastly more infectious than planktonic (free-living) cells. Here, we sought to identify factors that contribute to biofilm-induced hyperinfectivity in V. cholerae, and we present evidence for one aspect of the molecular basis of this phenotype. We identified proteins upregulated during growth in biofilms and determined their contributions to the hyperinfectivity phenotype. We found that PstS2, the periplasmic component of the Pst2 phosphate uptake system, was enriched in biofilms. Another gene in the pst2 locus was transcriptionally upregulated in biofilms. Using the infant mouse model, we found that mutation of two pst2 components resulted in impaired colonization. Importantly, deletion of the Pst2 inner membrane complex caused a greater colonization defect after growth in a biofilm compared to shaking culture. Based on these data, we propose that V. cholerae cells in biofilms upregulate the Pst2 system and therefore gain an advantage upon entry into the host. Further characterization of factors contributing to biofilm-induced hyperinfectivity in V. cholerae will improve our understanding of the transmission of the bacteria from natural aquatic habitats to the human host.
Continuous monitoring reveals multiple controls on ecosystem metabolism in a suburban stream

SUMMARY 1. Primary production and respiration in streams, collectively referred to as stream ecosystem metabolism, are fundamental processes that determine trophic structure, biomass and nutrient cycling. Few studies have used high-frequency measurements of gross primary production (GPP) and ecosystem respiration (ER) over extended periods to characterise the factors that control stream ecosystem metabolism at hourly, daily, seasonal and annual scales. 2. We measured ecosystem metabolism at 5-min intervals for 23 months in Shepherd Creek, a small suburban stream in Cincinnati, Ohio (U.S.A.). 3. Daily GPP was best predicted by a model containing light and its synergistic interaction with water temperature. Water temperature alone was not significantly related to daily GPP, rather high temperatures enhanced the capacity of autotrophs to use available light. 4. The relationship between GPP and light was further explored using photosynthesis–irradiance curves (P–I curves). Light saturation of GPP was evident throughout the winter and spring and the P–I curve frequently exhibited strong counterclockwise hysteresis. Hysteresis occurred when water temperatures were greater in the afternoon than in the morning, although light was similar, further suggesting that light availability interacts synergistically with water temperature. 5. Storm flows strongly depressed GPP in the spring while desiccation arrested aquatic GPP and ER in late summer and autumn. 6. Ecosystem respiration was best predicted by GPP, water temperature and the rate of water exchange between the surface channel and transient storage zones. We estimate that c. 70% of newly fixed carbon was immediately respired by autotrophs and closely associated heterotrophs. 7. Interannual, seasonal, daily and hourly variability in ecosystem metabolism was attributable to a combination of light availability, water temperature, storm flow dynamics and desiccation. Human activities affect all these factors in urban and suburban streams, suggesting stream ecosystem processes are likely to respond in complex ways to changing land use and climate.
An Arcanobacterium(Actinomyces) pyogenes Mutant Deficient in Production of the Pore-Forming Cytolysin Pyolysin Has Reduced Virulence

ABSTRACT Pyolysin (PLO), the hemolytic exotoxin expressed byArcanobacterium (Actinomyces)pyogenes, is a member of the thiol-activated cytolysin family of bacterial toxins. Insertional inactivation of theplo gene results in loss of expression of PLO with a concomitant loss in hemolytic activity. The plo mutant, PLO-1, has an approximately 1.8-log10 reduction in the 50% infectious dose compared to that for wild-type A. pyogenesin a mouse intraperitoneal infection model. Studies involving cochallenge of wild-type and PLO-1 bacteria resulted in recovery of similar numbers of both strains, suggesting that PLO production is required for survival in vivo. Recombinant, His-tagged PLO (His-PLO) is cytotoxic for mouse peritoneal macrophages and J774 cells in a dose-dependent manner. Protection against challenge with A. pyogenes could be afforded by vaccination with formalin-inactivated His-PLO, suggesting that PLO is a host-protective antigen, as well as a virulence determinant.
Keratinocyte growth factor and the transcription factors C/EBPα, C/EBPδ, and SREBP-1c regulate fatty acid synthesis in alveolar type II cells

Strategies to stimulate endogenous surfactant production require a detailed understanding of the regulation of lipogenesis in alveolar type II cells. We developed culture conditions in which keratinocyte growth factor (KGF) stimulates fatty acid and phospholipid synthesis. KGF stimulated acetate incorporation into phosphatidylcholine, disaturated phosphatidylcholin, and phosphatidylglycerol more than 5% rat serum alone. To determine the mRNA levels of lipogenic enzymes and transport proteins, we analyzed gene expression by oligonucleotide microarrays. KGF increased the mRNA levels for fatty acid synthase, stearoyl-CoA desaturase-1 (SCD-1), and epidermal fatty acid–binding protein more than rat serum alone. In addition, KGF increased the mRNA levels of the transcription factors CCAAT/enhancer-binding protein α (C/EBPα) and C/EBPδ as well as SREBP-1c (ADD-1), but not PPARγ. These changes in C/EBPα and C/EBPδ were confirmed by in situ hybridization. SCD-1 was also found to be highly expressed in alveolar type II cells in vivo. Furthermore, KGF increased protein levels of fatty acid synthase, C/EBPα, C/EBPδ, SREBP-1, epidermal fatty acid–binding protein, and SCD. Finally, the liver X receptor agonist T0901317 increased acetate incorporation and SREBP-1 but not SREBP-2 protein levels. In summary, KGF stimulates lipogenesis in type II cells by a coordinated expression of lipogenic enzymes and transport proteins regulated by C/EBP isoforms and SREBP-1c.
Expression of Organic Anion Transporting Polypeptides (OATPs) in Cancer Cell Lines and Tissues

Das Ovarialkarzinom ist eine der haufigsten Krebsarten im weiblichen Genitaltrakt mit der schlechtesten Prognose aller gynakologischen Erkrankungen. Die Ursache dafur liegt in der spaten Diagnose und der Chemoresistenz. Ein sehr groses Problem dabei ist, dass prognostizierende Faktoren und Tumormarker, vor allem CA-125, nicht spezifisch genug sind um eine fruhe Diagnose in Stadien, in denen sich der Tumor noch nicht uber die Ovarien hinaus verbreitet hat, zu ermoglichen. Das „Ewing’s Sarcoma” ist ein kleinzelliger Tumor, der vorwiegend in den Knochen von Kindern lokalisiert ist. Er zeichnet sich durch hohe Expression von CD99 in den Zellmembranen aus und in 85% der Falle durch eine reziproke Translokation des Chromosoms 11 des EWS Gens und des Chromosoms 22 des fli1 Gens. Diese Translokation resultiert in der Bildung eines chimaren EWS-fli1 Fusionsproteins. In beiden Krebsarten ist eine Verbesserung der Therapie wunschenswert. Diese konnte durch eine fruhe Diagnose mit geeigneten Tumormarkern und einer Erhohung der Effektivitat der Standard-Zytostatika erreicht werden. Deshalb konnten moglicherweise Transportproteine der Familie der OATPs, verantwortlich fur den Transport verschiedener Zytostatika, neue Therapieansatze liefern. Untersuchungen von OATPs scheinen daher eine vielversprechende Moglichkeit zu sein neue Angriffsziele in der Therapie zu entwickeln, um die Prognose fur diese Krebsarten zu verbessern. Daher war es Ziel dieser Diplomarbeit die Verteilung aller 11 bekannten humanen OATPs, durch einen Vergleich ihres Expressionsmusters in Krebszellen und in gesundem Gewebe, zu analysieren. „Quantitative real time RT-PCR” wurde verwendet um die Expression aller 11 bekannten humanen OATPs im „Ewing´s Sarcoma“ zu analysieren. Expressionsdaten wurden mit gesundem Gewebe und kolorektalen Krebszelllinien verglichen, die beide als Kontrolle dienten. Weiters wurden Ovarialkrebszellen mit den Genen OATP1B1 und OATP1B3 transfeziert um diese anschliesend verschiedenen Konzentrationen des zytotoxischen Agens, Paclitaxel, auszusetzen. In den „Quantitativen real time RT-PCR” Experimenten zeigte die „Ewing’s Sarcoma“ Zelllinie TC-71 eine signifikant hohere Expressionsrate des OATP5A1-Gens verglichen mit normalem Gehirngewebe. Hinsichtlich des Zytotoxizitatstests waren Zellen der Paclitaxel-sensitiven Ovarialkarzinomzelllinie SK-OV-3 nach Transfektion mit den Genen OATP1B1 und OATP1B3 noch sensitiver gegenuber Paclitaxel, gemessen an niedrigeren IC50-Werten. Zusammenfassend bestatigen diese Daten eine signifikante Expression von OATPs in beiden Krebsarten und ihre Zelllinien bieten sich als Model fur weitere Zytostatikastudien an.
Biological Properties of Structurally Related α-Helical Cationic Antimicrobial Peptides

ABSTRACT A series of α-helical cationic antimicrobial peptide variants with small amino acid changes was designed. Alterations in the charge, hydrophobicity, or length of the variant peptides did not improve the antimicrobial activity, and there was no statistically significant correlation between any of these factors and the MIC forPseudomonas aeruginosa, Escherichia coli, orSalmonella typhimurium. Individual peptides demonstrated synergy with conventional antibiotics against antibiotic-resistant strains of P. aeruginosa. The peptides varied considerably in the ability to bind E. coli O111:B4 lipopolysaccharide (LPS), and this correlated significantly with their antimicrobial activity and ability to block LPS-stimulated tumor necrosis factor and interleukin-6 production. In general, the peptides studied here demonstrated a broad range of activities, including antimicrobial, antiendotoxin, and enhancer activities.
Fostering Conservation via an Integrated Use of Conventional Approaches and High-Throughput SPET Genotyping: A Case Study Using the Endangered Canarian Endemics Solanum lidii and S. vespertilio (Solanaceae)

Islands provide unique opportunities to integrated research approaches to study evolution and conservation because boundaries are circumscribed, geological ages are often precise, and many taxa are greatly imperiled. We combined morphological and hybridization studies with high-throughput genotyping platforms to streamline relationships in the endangered monophyletic and highly diverse lineage of Solanum in the Canarian archipelago, where three endemic taxa are currently recognized. Inter-taxa hybridizations were performed, and morphological expression was assessed with a common-garden approach. Using the eggplant Single Primer Enrichment Technology (SPET) platform with 5,093 probes, 74 individuals of three endemic taxa (Solanum lidii, S. vespertilio subsp. vespertilio, and S. vespertilio subsp. doramae) were sampled for SNPs. While morphological and breeding studies showed clear distinctions and some continuous variation, inter-taxon hybrids were fertile and heterotic for vigor traits. SPET genotyping revealed 1,421 high-quality SNPs and supported four, not three, distinct taxonomic entities associated with post-emergence geological, ecological and geographic factors of the islands. Given the lack of barriers to hybridization among all the taxa and their molecular differences, great care must be taken in population management. Conservation strategies must take account of the sexual and breeding systems and genotypic distribution among populations to successfully conserve and restore threatened/endangered island taxa, as exemplified by Solanum on the Canary Islands.
Engineered Bacterial Outer Membrane Vesicles as Multifunctional Delivery Platforms

Outer membrane vesicles (OMVs) are valued for their unique, convenient, and amendable functions. They are flexible, controllable nanoparticles, which can be modified in different ways, for use in a wide array of applications like adjuvants, vaccines, drug presentation, and fluorescence tracking. Attempts have been made to alter the nanoparticles in liposomes and bacterial ghosts using traditional methods. However, it has not been highly successful owing to certain unavoidable disadvantages. OMVs have unique advantages and are considered novel platforms for heterologous protein presentation. Although previous reviews have described this to some extent, owing to the rapid development in this field, there is a need for the regular update of the reviews. This review focuses on few novel applications of engineered OMVs, and redefines the significance of OMVs as novel multifunctional delivery platforms. Additionally, this review also supplements and provides an update on the OMV transformation methods.
Vegetative Propagation of Mature Teak Trees (Tectona grandis L.)

Summary Clonal propagation technology for mature teak trees (63 year-old) has been achieved for the first time. Indole butyric acid (IBA) was found to be the most effective auxin tested. Cuttings from coppice shoots of mature trees rooted between 74% to 91% with a 2000 ppm IBA treatment, while the cuttings from 1 to 2 year-old root-stocks (stumps) rooted between 79% to 100% with 1000ppm IBA applied at different times of the year. Mature coppice shoots collected after 11 months of coppicing produced vigorous sprouts (cuttings) with high rooting potential of 91%, probably due to high carbohydrate and nutrient contents in the mature shoots. Adventitious root formation and root system of cuttings collected from planted coppice shoots of 63 year-old mature trees were similar to those of one year-old root-stocks, suggesting the juvenility of coppice shoots regarding rooting. In teak, rooting depends upon the physiological status of the cutting, and season is no longer a major barrier in rhizogenesis.
Structure and Organization of Effective Peanut and Cowpea Root Nodules Induced by Rhizobial Strain 32H1

Sen, D. , Weaver, R. W. and Bal, A. K. 1986. Structure and organization of effective peanut and cowpea root nodules induced by rhizobial strain 32H1. —J. exp. Bot. 37: 356-363. A comparison of the structure and organization of nodular tissues and bacteroids of peanut and cowpea induced by Rhizobium sp. strain 32H1 was made 4 to 5 weeks after inoculation when nitrogenase activity reaches the peak. Observations revealed major differences that may have a role in the different rates of nitrogen fixation shown by the two species. All cell types in cowpea nodules were larger than those of peanut. The inner cortex of cowpea had an 'endodermis-like' layer of cells which was absent in peanut. All cells in the bacteroidal zone of peanut were infected but in cowpea many remained free of bacteroids. The bacteroid containing cells of peanut were isodiametrical and uniform in size with a central vacuole and a nucleus surrounded by tightly arranged bacteroids enclosed singly in peribacteroidal membrane sacs. Such cells in cowpea were mostly elongated with a nucleus and one or more vacuoles. The bacteroids within cowpea cells were arranged without any particular order with more space for host cellular material. They were mostly present singly in peribacteroidal membrane sacs which sometimes fused to enclose more than one bacteroid. The hosts seem to play the dominant role in the differentiation of nodular tissue and the morphogenesis of bacteroids in symbiotic systems induced by the same strain of Rhizobium. Key words—Peanut, cowpea, nodule structure. Correspondence to: Department of Soil and Crop Sciences, Texas Agricultural Experiment Station and Texas A. & M. University, College Station, Texas, U.S.A. 77843.
A Plaque Assay for Nuclear Polyhedrosis Viruses using a Solid Overlay

Summary The nuclear polyhedrosis viruses of Trichoplusia ni and Autographa californica produce plaques in monolayers of Spodoptera frugiperda cells under a solid overlay containing agarose as the solidifying agent. Plaques are visible macroscopically after staining the cells with neutral red or with the tetrazolium salt, INT, and a linear dose response is observed. The sensitivity of the assay is less than that obtained using an end-point dilution technique; however, plaque formation does provide a simple means of cloning virus.
Bacterial leaf glands in Styrax camporum (Styracaceae): first report for the family

In this study, we recorded, for the first time, the occurrence of leaf glands in a member of Styracaceae and their association with bacteria. Samples of Styrax camporum Pohl shoot apices and leaves at different developmental stages were prepared according to the conventional techniques for light and electron microscopy. Glands are emergences constituted by epidermal and parenchyma cells and are differentiated into a secretory body on a short nonsecretory stalk supplied with phloem. Actively secreting glands occur from leaf primordia to mature leaves and produce mucilage that accumulates inside schizogenous intercellular spaces. The epidermal secretory cells have abundant cytoplasm rich in hyperactive dictyosomes, an extensive endoplasmic reticulum, and modified plastids. Bacteria enter the gland via the intact surface and proliferate in the intercellular spaces of the glands. Once inside the intercellular spaces of the glands, bacteria enter the cells owing to the weakening of the anticlinal and inner per.
UPQC System Configuration for Single Phase and Three Phase Network : A Review

This paper presents a literature review on the broad overview on the different possible UPQC system configurations for single-phase (two-wire) and three-phase (three-wire and four-wire) networks, different compensation approaches, and recent developments to enhance the electric power quality at distribution levels. Therefore, a list on the types of different UPQC configuration is developed and presented to highlight the distinguishing feature offered by a particular UPQC.
An ex vivo Approach to Study Hormonal Control of Spermatogenesis in the Teleost Oreochromis niloticus

As the male reproductive organ, the main task of the testis is the production of fertile, haploid spermatozoa. This process, named spermatogenesis, starts with spermatogonial stem cells, which undergo a species-specific number of mitotic divisions until starting meiosis and further morphological maturation. The pituitary gonadotropins, luteinizing hormone, and follicle stimulating hormone, are indispensable for vertebrate spermatogenesis, but we are still far from fully understanding the complex regulatory networks involved in this process. Therefore, we developed an ex vivo testis cultivation system which allows evaluating the occurring changes in histology and gene expression. The experimental circulatory flow-through setup described in this work provides the possibility to study the function of the male tilapia gonads on a cellular and transcriptional level for at least 7 days. After 1 week of culture, tilapia testis slices kept their structure and all stages of spermatogenesis could be detected histologically. Without pituitary extract (tilPE) however, fibrotic structures appeared, whereas addition of tilPE preserved spermatogenic cysts and somatic interstitium completely. We could show that tilPE has a stimulatory effect on spermatogonia proliferation in our culture system. In the presence of tilPE or hCG, the gene expression of steroidogenesis related genes (cyp11b2 and stAR2) were notably increased. Other testicular genes like piwil1, amh, or dmrt1 were not expressed differentially in the presence or absence of gonadotropins or gonadotropin containing tilPE. We established a suitable system for studying tilapia spermatogenesis ex vivo with promise for future applications.
The Alternative Sigma Factor, ςE, Is Critically Important for the Virulence of Salmonella typhimurium

ABSTRACT In Escherichia coli, extracytoplasmic stress is partially controlled by the alternative sigma factor, RpoE (ςE). In response to environmental stress or alteration in the protein content of the cell envelope, ςEupregulates the expression of a number of genes, includinghtrA. It has been shown that htrA is required for intramacrophage survival and virulence in Salmonella typhimurium. To investigate whether ςE-regulated genes other than htrA are involved in salmonella virulence, we inactivated the rpoE gene of S. typhimurium SL1344 by allelic exchange and compared the phenotype of the mutant (GVB311) in vitro and in vivo with its parent and an isogenic htrA mutant (BRD915). UnlikeE. coli, ςE is not required for the growth and survival of S. typhimurium at high temperatures. However, GVB311 did display a defect in its ability to utilize carbon sources other than glucose. GVB311 was more sensitive to hydrogen peroxide, superoxide, and antimicrobial peptides than SL1344 and BRD915. Although able to invade both macrophage and epithelial cell lines normally, the rpoE mutant was defective in its ability to survive and proliferate in both cell lines. The effect of the rpoE mutation on the intracellular behavior of S. typhimurium was greater than that of the htrA mutation. Both GVB311 and BRD915 were highly attenuated in mice. Neither strain was able to kill mice via the oral route, and the 50% lethal dose (LD50) for both strains via the intravenous (i.v.) route was very high. The i.v. LD50s for SL1344, BRD915, and GVB311 were <10, 5.5 × 105, and 1.24 × 107 CFU, respectively. Growth in murine tissues after oral and i.v. inoculation was impaired for both thehtrA and rpoE mutant, with the latter mutant being more severely affected. Neither mutant was able to translocate successfully from the Peyer’s patches to other organs after oral infection or to proliferate in the liver and spleen after i.v. inoculation. However, the htrA mutant efficiently colonized the livers and spleens of mice infected i.v. , but the rpoE mutant did not. Previous studies have shown that salmonella htrA mutants are excellent live vaccines. In contrast, oral immunization of mice with GVB311 was unable to protect any of the mice from oral challenge with SL1344. Furthermore, i.v. immunization with a large dose (∼106 CFU) of GVB311 protected less than half of the orally challenged mice. Thus, our results indicate that genes in the ςE regulon other than htrA play a critical role in the virulence and immunogenicity of S. typhimurium.