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Query: "author" (Michael P. O'Connell) .

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1.
The digital MIQE guidelines update : minimum information for publication of quantitative digital PCR experiments for 2020
Jim F. Huggett, Alexandra S. Whale, Ward De Spiegelaere, Afif M. Abdel Nour, Young-Kyung Bae, Vladimír Beneš, Dan Burke, Megan Cleveland, Philippe Corbisier, Alison S. Devonshire, Lianhua Dong, Daniela Drandi, Carole A. Foy, Jeremy A. Garson, Hua-Jun He, Jan Hellemans, Mikael Kubista, Antoon Lievens, Mike G. Makrigiorgos, Mojca Milavec, Reinhold D. Mueller, Tania Nolan, Denise M. O'Sullivan, Michael W. Pfaffl, Stefan Rödiger, Erica L. Romsos, Gregory L. Shipley, Valérie Taly, Andreas Untergasser, Carl T. Wittwer, Stephen A. Bustin, Jo Vandesompele, 2020, review article

Abstract: Digital PCR (dPCR) has developed considerably since the publication of the Minimum Information for Publication of Digital PCR Experiments (dMIQE) guidelines in 2013, with advances in instrumentation, software, applications, and our understanding of its technological potential. Yet these developments also have associated challenges; data analysis steps, including threshold setting, can be difficult and preanalytical steps required to purify, concentrate, and modify nucleic acids can lead to measurement error. To assist independent corroboration of conclusions, comprehensive disclosure of all relevant experimental details is required. To support the community and reflect the growing use of dPCR, we present an update to dMIQE, dMIQE2020, including a simplified dMIQE table format to assist researchers in providing key experimental information and understanding of the associated experimental process. Adoption of dMIQE2020 by the scientific community will assist in standardizing experimental protocols, maximize efficient utilization of resources, and further enhance the impact of this powerful technology.
Published in DiRROS: 06.08.2024; Views: 27; Downloads: 77
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2.
CellTracker Green labelling vs. Rose Bengal staining: CTG wins by points in distinguishing living from dead anoxia-impacted copepods and nematods
Mateja Grego, Michael Stachowitsch, Marleen De Troch, Bettina Riedel, 2013, original scientific article

Abstract: Hypoxia and anoxia have become a key threat to shallow coastal seas. Much is known about their impact on macrofauna, less on meiofauna. In an attempt to shed more light on the latter group, in particular from a process-oriented view, we experimentally induced short-term anoxia (1 week) in the northern Adriatic Sea (Mediterranean) and examined the two most abundant meiofauna taxa – harpacticoid copepods and nematodes. Both taxa also represent different ends of the tolerance spectrum, with copepods being the most sensitive and nematodes among the most tolerant. We compared two methods: CellTracker Green (CTG) – new labelling approach for meiofauna – with the traditional rose bengal (RB) staining method. CTG binds to active enzymes and therefore colours live organisms only. The two methods show considerable differences in the number of living and dead individuals of both meiofauna taxa. Generally, RB will stain dead but not yet decomposed copepods and nematodes equally as it does live ones. Specifically, RB significantly overestimated the number of living copepods in all sediment layers in anoxic samples, but not in any normoxic samples. In contrast, for nematodes, the methods did not show such a clear difference between anoxia and normoxia. RB overestimated the number of living nematodes in the top sediment layer of normoxic samples, which implies an overestimation of the overall live nematofauna. For monitoring and biodiversity studies, the RB method might be sufficient, but for more precise quantification of community degradation, especially after an oxygen depletion event, CTG labelling is a better tool. Moreover, it clearly highlights the surviving species within the copepod or nematode community. As already accepted for foraminiferal research, we demonstrate that the CTG labelling is also valid for other meiofauna groups.
Keywords: morje, plankton, kopepodi, živalski plankton, trofični status, morski plankton, CTG, CellTracker Green
Published in DiRROS: 02.08.2024; Views: 73; Downloads: 77
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3.
TRIM28 and [beta]-actin identified via nanobody-based reverse proteomics approach as possible human glioblastoma biomarkers
Ivana Jovchevska, Neja Šamec, Nina Kočevar Britovšek, Daniela Cesselli, Neža Podergajs, Clara Limbaeck Stanic, Michael P. Myers, Serge Muyldermans, Gholamreza Hassanzadeh Ghassabeh, Helena Motaln, Maria Elisabetta Ruaro, Evgenia Bourkoula, Tamara Lah Turnšek, Radovan Komel, 2014, original scientific article

Abstract: Malignant gliomas are among the rarest brain tumours, and they have the worst prognosis. Grade IV astrocytoma, known as glioblastoma multiforme (GBM), is a highly lethal disease where the standard therapies of surgery, followed by radiation and chemotherapy, cannot significantly prolong the life expectancy of the patients. Tumour recurrence shows more aggressive form compared to the primary tumour, and results in patient survival from 12 to 15 months only. Although still controversial, the cancer stem cell hypothesis postulates that cancer stem cells are responsible for early relapse of the disease after surgical intervention due to their high resistance to therapy. Alternative strategies for GBM therapy are thus urgently needed. Nanobodies are single-domain antigen-binding fragments of heavy-chain antibodies, and together with classical antibodies, they are part of the camelid immune system. Nanobodies are small and stable, and they share a high degree of sequence identity to the human heavy chain variable domain, and these characteristics offer them advantages over classical antibodies or antibody fragments. We first immunised an alpaca with a human GBM stem-like cell line prepared from primary GBM cultures. Next, a nanobody library was constructed in a phage-display vector. Using nanobody phage-display technology, we selected specific GBM stem-like cell binders through a number of affinity selections, using whole cell protein extracts and membrane protein-enriched extracts from eight different GBM patients, and membrane protein-enriched extracts from two established GBM stem-like cell lines (NCH644 and NCH421K cells). After the enrichment, periplasmic extract ELISA was used to screen for specific clones. These nanobody clones were recloned into the pHEN6 vector, expressed in Escherichia coli WK6, and purified using immobilised metal affinity chromatography and size-exclusion chromatography. Specific nanobody:antigen pairs were obtained and mass spectrometry analysis revealed two proteins, TRIM28 and β-actin, that were up-regulated in the GBM stem-like cells compared to the controls.
Keywords: malignant gliomas, cancer stem cells, nanobodies
Published in DiRROS: 02.08.2024; Views: 89; Downloads: 68
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4.
Artificially induced migration of redox layers in a coastal sediment from the Northern Adriatic
Eduard Metzger, D. Langlet, E. Viollier, Neža Koron, Bettina Riedel, Michael Stachowitsch, Jadran Faganeli, M. Tharaud, E. Geslin, F. Jorissen, 2014, original scientific article

Abstract: Long-term experimental studies suggest that, under transient anoxic conditions, redox fronts within the sediment shift upwards, causing sequential rise and fall of benthic fluxes of reduced species (Mn(II), Fe(II) and S(-II)). Infaunal benthic organisms are associated with different redox fronts as micro-habitats and must be affected by such changes during natural hypoxia events. In order to document the geochemical evolution of the sediment during prolonged anoxia in the framework of an in situ experiment designed to mimic natural conditions, benthic chambers were deployed on the seafloor of the Northern Adriatic and sampled after 9, 30 and 315 days of incubation. Oxygen and sulfide were measured continuously in the early stages (9 days) of the experiment. High-resolution pore water profiles were sampled by DET probes and redox-sensitive species (S(VI), Mn(II) and Fe(II)) and alkalinity were measured. Starting oxygen saturation was about 80% within the chamber. After 7 days, anoxia was established in the bottom waters within the chambers. Mn(II) and Fe(II) started diffusing towards the anoxic water column until they reached the surficial sediment. Being reoxidized there, Mn and Fe reprecipitated, giving a rusty coloration to the seafloor. Infaunal species appeared at the sediment surface. After 20 days, all macro-organisms were dead. Decomposition of macro-organisms at the sediment–water interface generated S(-II) within the entire height of the chamber, leading to a downward flux of sulfides into the sediment, where they were quickly oxidized by metallic oxides or precipitated as FeS. S(-II) was below detection in the water column and pore waters at the end of the experiment. Our results suggest that S(-II) enrichment in the water column of coastal systems, which are episodically anoxic, is strongly controlled by the biomass of benthic macrofauna and its decay during anoxia, whereas its residence time in the water column is controlled by iron availability (as solid oxides or as dissolved reduced cations) within the sediment, even without water circulation.
Keywords: Adriatic Sea, marine sediments, anoxia, Northern Adriatic
Published in DiRROS: 02.08.2024; Views: 62; Downloads: 85
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5.
Meiofauna winners and losers of coastal hypoxia: case study harpacticoid copepods
Mateja Grego, Bettina Riedel, Michael Stachowitsch, Marleen De Troch, 2014, original scientific article

Abstract: The impact of anoxia on meiobenthic copepod species was assessed by means of a field experiment. Four plexiglass chambers were deployed in situ in 24 m depth to simulate an anoxic event of 9 days, 1 month, 2 months and 10 months. From normoxic to anoxic conditions, we recorded a drop in copepod density and species richness. With increasing duration of anoxia the relative abundance of the individuals of the family Cletodidae increased, and they survived the 1 month and 2 month anoxia, the latter with few specimens. They were the true "winners" of the experimentally induced anoxia. Dominance did not increase in the deployments because not one, but several species from this family were tolerant to anoxia. The overall rate of survival was the same for males and females, but no juvenile stages of copepods survived in anoxia. During a recovery phase of 7 days after a short-term anoxia of 9 days, harpacticoid copepod density did not increase significantly, and there was only a slight increase in species diversity. We concluded that no substantial colonisation from the surrounding sediment took place. The survivors, however, showed a high potential for recovery according to the number of gravid females, whose number increased significantly once the oxygen was available again. These findings imply that substantial energy is allocated to reproduction in the recovery phase.
Published in DiRROS: 01.08.2024; Views: 93; Downloads: 59
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6.
NanoSIMS and tissue autoradiography reveal symbiont carbon fixation and organic carbon transfer to giant ciliate host
Jean-Marie Volland, Arno Schintlmeister, Helena Zambalos, Siegfried Reipert, Patricija Mozetič, Salvador Espada-Hinojosa, Valentina Turk, Michael Wagner, Monika Bright, 2018, original scientific article

Abstract: The giant colonial ciliate Zoothamnium niveum harbors a monolayer of the gammaproteobacteria Cand. Thiobios zoothamnicoli on its outer surface. Cultivation experiments revealed maximal growth and survival under steady flow of high oxygen and low sulfide concentrations. We aimed at directly demonstrating the sulfur-oxidizing, chemoautotrophic nature of the symbionts and at investigating putative carbon transfer from the symbiont to the ciliate host. We performed pulse-chase incubations with 14C- and 13C-labeled bicarbonate under varying environmental conditions. A combination of tissue autoradiography and nanoscale secondary ion mass spectrometry coupled with transmission electron microscopy was used to follow the fate of the radioactive and stable isotopes of carbon, respectively. We show that symbiont cells fix substantial amounts of inorganic carbon in the presence of sulfide, but also (to a lesser degree) in the absence of sulfide by utilizing internally stored sulfur. Isotope labeling patterns point to translocation of organic carbon to the host through both release of these compounds and digestion of symbiont cells. The latter mechanism is also supported by ultracytochemical detection of acid phosphatase in lysosomes and in food vacuoles of ciliate cells. Fluorescence in situ hybridization of freshly collected ciliates revealed that the vast majority of ingested microbial cells were ectosymbionts.
Keywords: microbial ecology, symbiosis
Published in DiRROS: 24.07.2024; Views: 88; Downloads: 101
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7.
New alien Mediterranean biodiversity records 2020
Michel Bariche, Sara A. A. Al-Mabruk, Maria A. Ateş, Adnan Büyük, Fabio Crocetta, Michail Dritsas, Diala Edde, Ana Fortič, Lovrenc Lipej, 2020, original scientific article

Abstract: The current article presents 18 new records from seven Mediterranean countries. These records include one rhodophyte, four nudibranchs, two crustaceans, one stingray and 10 bony fishes. They are grouped by country as follows: Lebanon - first record of the Striped bass Morone saxatilis, the stingray Himantura leoparda, the Areolate grouper Epinephelus areolatus and the Spot-fin porcupinefish Diodon hystrix from various parts of the country; Turkey - first record of the invasive red alga Grateloupia turuturu from the sea of Marmara (region of Bandırma), the sea slug Goniobranchus obsoletus and the crab Arcania brevifrons from the Gulf of Antalya and the cladoceran Pleopis schmackeri from several locations along the Aegean Sea; Cyprus - first record of the alien sea slug Berthellina citrina from the region of Cape Greco and an observation of a butterflyfish Heniochus sp. from the northeastern side of the island; Greece - first record of the alien sea slug Anteaeolidiella lurana from the region of Heraklion in Crete and the record of the Atlantic spadefish Chaetodipterus faber and the Black surgeonfish Acanthurus cfr gahhm from Salamina Island; Slovenia - first record of the alien sea slug Thecacera pennigera from Izola; Italy - first record of the hybrid Striped bass (Morone saxatilis × Morone chrysops) from the northern Tyrrhenian Sea and a first record of the goldfish Carassius auratus from the region of Apulia; Libya - first record of the Red Sea goatfish Parupeneus forsskali and the African surgeonfish Acanthurus monroviae, respectively from the eastern (Al-Tamimi area) and the western shore (Al-Khums area).
Published in DiRROS: 23.07.2024; Views: 227; Downloads: 100
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8.
Genotype-environment interactions rule the response of a widespread butterfly to temperature variation
Franziska Günter, Michaël Beaulieu, Kasimir F. Freiberg, Ines Welzel, Nia Toshkova, Anamarija Žagar, Tatjana Simčič, Klaus Fischer, 2020, original scientific article

Abstract: Understanding how organisms adapt to complex environments is a central goal of evolutionary biology and ecology. This issue is of special interest in the current era of rapidly changing climatic conditions. Here, we investigate clinal variation and plastic responses in life history, morphology and physiology in the butterfly Pieris napi along a pan‐European gradient by exposing butterflies raised in captivity to different temperatures. We found clinal variation in body size, growth rates and concomitant development time, wing aspect ratio, wing melanization and heat tolerance. Individuals from warmer environments were more heat‐tolerant and had less melanised wings and a shorter development, but still they were larger than individuals from cooler environments. These findings suggest selection for rapid growth in the warmth and for wing melanization in the cold, and thus fine‐tuned genetic adaptation to local climates. Irrespective of the origin of butterflies, the effects of higher developmental temperature were largely as expected, speeding up development; reducing body size, potential metabolic activity and wing melanization; while increasing heat tolerance. At least in part, these patterns likely reflect adaptive phenotypic plasticity. In summary, our study revealed pronounced plastic and genetic responses, which may indicate high adaptive capacities in our study organism. Whether this may help such species, though, to deal with current climate change needs further investigation, as clinal patterns have typically evolved over long periods.
Published in DiRROS: 23.07.2024; Views: 75; Downloads: 40
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9.
New alien Mediterranean biodiversity records (October 2020)
Michail Ragkousis, Nardjes Abdelali, Ernesto Azzurro, Ali Badreddine, Michel Bariche, Ghazi Bitar, Fabio Crocetta, Francesco Denitto, Ana Fortič, Borut Mavrič, 2020, original scientific article

Abstract: This article includes 23 new records of alien and cryptogenic species in the Mediterranean Sea, belonging to 4 Phyla (Chordata, Echinodermata, Arthropoda and Mollusca), distributed from the Alboran to the Levantine Sea. Records are reported from eight countries listed from West to East as follows: Algeria: new records of the Atlantic blue crab Callinectes sapidus; Spain: further spread and establishment of the sea slug Lamprohaminoea ovalis in continental shores; Tunisia: first record of the Atlantic Blue Crab Callinectes sapidus in the Gulf of Gabes; Italy: a new occurrence of the pufferfish Lagocephalus sceleratus in Northern Ionian waters; first record of Cephalopholis taeniops in the Ionian Sea; first record of the redlip blenny, Ophioblennius atlanticus in the Ionian Sea; Slovenia: first record of the isopod Paranthura japonica in Slovenia; Greece: first record of the molluscs Eunaticina papilla, Plocamopherus ocellatus and the fish Cheilodipterus novemstriatus; first record of the ascidian Ecteinascidia turbinata in Kriti; the long-spined sea urchin Diadema setosum in the Ionian Sea; Turkey: first record of the sea spider Ammothea hilgendorfi; the stomatopod Cloridina cf. ichneumon; the fishes Pempheris rhomboidea from the Sea of Marmara and Paranthias furcifer from the Aegean Sea; Lebanon: new records of the fishes Arothron hispidus, Rachycentron canadum, Heniochus intermedius and Acanthurus monroviae; first record of Acanthostracion polygonius. The records of Cloridina cf. ichneumon from southern Turkey and the fish Acanthostracion polygonius from Lebanon, both being the first Mediterranean records, are noteworthy.
Published in DiRROS: 22.07.2024; Views: 233; Downloads: 100
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10.
Evidence for senescence in survival but not in reproduction in a short-lived passerine
Rémi Fay, Michael Schaub, Jennifer A. Border, Ian G. Henderson, Georg Fahl, Jürgen Feulner, Petra Horch, Mathis Müller, Helmut Rebstock, Dmitry Shitikov, Davorin Tome, Matthias Vögeli, Martin U. Grüebler, 2020, original scientific article

Abstract: Senescence has been studied since a long time by theoreticians in ecology and evolution, but empirical support in natural population has only recently been accumulating. One of the current challenges is the investigation of senescence of multiple fitness components and the study of differences between sexes. Until now, studies have been more frequently conducted on females than on males and rather in long-lived than in short-lived species. To reach a more fundamental understanding of the evolution of senescence, it is critical to investigate age-specific survival and reproduction performance in both sexes and in a large range of species with contrasting life histories. In this study, we present results on patterns of age-specific and sex-specific variation in survival and reproduction in the whinchat Saxicola rubetra, a short-lived passerine. We compiled individual-based long-term datasets from seven populations that were jointly analyzed within a Bayesian modeling framework. We found evidence for senescence in survival with a continuous decline after the age of 1 year, but no evidence of reproductive senescence. Furthermore, we found no clear evidence for sex effects on these patterns. We discuss these results in light of previous studies documenting senescence in short-lived birds. We note that most of them have been conducted in populations breeding in nest boxes, and we question the potential effect of the nest boxes on the shape of age-reproductive trajectories.
Keywords: actuarial senescence, age-specific demographic rate, aging, Saxicola rubetra, whinchat
Published in DiRROS: 22.07.2024; Views: 97; Downloads: 58
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