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Abstract: Jellyfish are becoming an increasingly important component of studies of the global marine environment, as their frequent mass phenomena affect ecosystem performance and have economic and health consequences. Recent research has focused primarily on mass occurrences of scyphozoans and ctenophores, while less attention has been paid to hydromedusae blooms, which can also have significant impacts on the marine ecosystem with economic consequences. Like many members of the Scyphozoa and Ctenophora, Aequorea species can be numerous and form blooms. Based on surveys from 2000 to 2020 and information provided by marine professionals, we examine the (multiannual) variability, seasonality, and spatial distribution patterns of the poorly studied temperate species Aequorea cf. forskalea (Hydrozoa: Leptothecata). We hypothesized that this macrogelatinous jellyfish with a bipartite life cycle (benthic polyps and planktonic medusae) is sensitive to seasonal and long-term temperature fluctuations, particularly because temperature has been shown to play a critical role in mediating transitions in the life cycle of cnidarians. The cold-water affinity of A. cf. forskalea was confirmed by a significant negative correlation with temperature and the most frequent occurrence during the winter-spring period. For the northern Adriatic (Gulf of Trieste), sea temperature data and semi-quantitative plankton data are available for the first decade of the 20th century, allowing comparison of the historical data (1900-1911) with our current data set. Compared to historical records, the current occurrence of A. cf. forskalea differs significantly. With the warming of the northern Adriatic Sea since the beginning of the 20th century, the current phenology of A. cf. forskalea has changed, probably related to the delayed production of medusae by hydroids in autumn, the absence of medusae in this season and their less frequent occurrence in winter.
Keywords: hydromedusae, blooms, long-term monitoring, multiannual variability, Mediterranean Sea
Published in DiRROS: 31.07.2024; Views: 561; Downloads: 178
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Abstract: Current methods based on DNA targets for the detection, identification and quantification of genetically modified organisms (GMOs) involve extraction of the DNA. Different extraction procedures have been developed for the great variety of samples from food, feed, seeds and particular plant parts. This makes the operation of routine analytical laboratories complex and workloads heavy. Here we present a decision-making system, developed over many years of GMO testing on different samples, that result in the application of only a few extraction methods for the majority of samples. Developed decision-making system enables quicker and more cost effective testing of GMOs. In addition, the performance of DNA extraction resulting from the use of the selected extraction methods is presented for use in subsequent testing of GMOs by real time PCR methods. This approach can be used as a model for similar systems based on nucleic acid analysis in food, feed, seeds and plants.
Keywords: extraction methods, genetically modified organisms, GMO, decision- making system (biology), GMO testing, cetyltrimethylammonium bromide, CTAB
Published in DiRROS: 29.07.2024; Views: 257; Downloads: 175
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Abstract: The Indo-Pacific egg-carrying copepod Pseudodiaptomus marinus Sato, 1913 was recorded for the first time in the Port of Koper, Slovenia (Gulf of Trieste) in February 2015. This is the fourth finding of this species in the Mediterranean Sea, and the third for the Adriatic Sea. A rather high abundance of 73 ind. m-3 was recorded, which is considerably higher than previous findings in the Adriatic. Maritime transport is presumed to be the main cause of its introduction, primarily through ballast water release. Repeat sampling in May confirmed the presence of this copepod, indicating the possibility that the species has established a stable population in the Port of Koper.
Keywords: copepods, Adriatic Sea, mesozooplankton, ballast waters
Published in DiRROS: 29.07.2024; Views: 270; Downloads: 97
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Abstract: Background Chinese hamster ovary (CHO) cells have become the host of choice for the production of recombinant proteins, due to their capacity for correct protein folding, assembly, and posttranslational modifications. The most widely used system for recombinant proteins is the gene amplification procedure that uses the CHO-Dhfr expression system. However, CHO cells are known to have a very unstable karyotype. This is due to chromosome rearrangements that can arise from translocations and homologous recombination, especially when cells with the CHO-Dhfr expression system are treated with methotrexate hydrate. The present method used in the industry for testing clones for their long-term stability of recombinant protein production is empirical, and it involves their cultivation over extended periods of time prior to the selection of the most suitable clone for further bioprocess development. The aim of the present study was the identification of marker genes that can predict stable expression of recombinant genes in particular clones early in the development stage. Results The transcriptome profiles of CHO clones with stable and unstable recombinant protein production were investigated over 10-weeks of cultivation, using a DNA microarray. We identified 14 genes that were differentially expressed between the stable and unstable clones already at 2 weeks from the beginning of the cultivation. Their expression was validated by reverse-transcription quantitative real-time PCR (RT-qPCR). Furthermore, the k-nearest neighbour algorithm approach shows that the combination of the gene expression patterns of only five of these 14 genes is sufficient to predict stable recombinant protein production in clones in the early phases of cell-line development. Conclusions The exact molecular mechanisms that cause unstable recombinant protein production are not fully understood. However, the expression profiles of some genes in clones with stable and unstable recombinant protein production allow prediction of such instability early in the cell-line development stage. We have thus developed a proof-of-concept for a novel approach to eliminate unstable clones in the CHO-Dhfr expression system, which saves time and labour-intensive work in cell-line development.
Keywords: CHO cell line, stable recombinant protein production, gene expression, RT-qPCR, DNA microarray, mMarker genes
Published in DiRROS: 29.07.2024; Views: 318; Downloads: 321
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Abstract: Glioblastoma multiforme is the most lethal of brain cancer, and it comprises a heterogeneous mixture of functionally distinct cancer cells that affect tumor progression. We examined the U87, U251, and U373 malignant cell lines as in vitro models to determine the impact of cellular cross-talk on their phenotypic alterations in co-cultures. These cells were also studied at the transcriptome level, to define the mechanisms of their observed mutually affected genomic stability, proliferation, invasion and resistance to temozolomide. This is the first direct demonstration of the neural and mesenchymal molecular fingerprints of U87 and U373 cells, respectively. U87-cell conditioned medium lowered the genomic stability of U373 (U251) cells, without affecting cell proliferation. In contrast, upon exposure of U87 cells to U373 (U251) conditioned medium, U87 cells showed increased genomic stability, decreased proliferation rates and increased invasion, due to a plethora of produced cytokines identified in the co-culture media. This cross talk altered the expression 264 genes in U87 cells that are associated with proliferation, inflammation, migration, and adhesion, and 221 genes in U373 cells that are associated with apoptosis, the cell cycle, cell differentiation and migration. Indirect and direct co-culturing of U87 and U373 cells showed mutually opposite effects on temozolomide resistance. In conclusion, definition of transcriptional alterations of distinct glioblastoma cells upon co-culturing provides better understanding of the mechanisms of glioblastoma heterogeneity, which will provide the basis for more informed glioma treatment in the future.
Keywords: glioblastoma heterogeneity, U87 cells, temozolomide resistance, cellular cross-talk, transcriptomics
Published in DiRROS: 29.07.2024; Views: 292; Downloads: 234
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