31. Stink bug communication and signal detection in a plant environmentAndrej Čokl, Alenka Žunič Kosi, Nataša Stritih Peljhan, Maria Carolina Blassioli Moraes, Raúl Alberto Laumann, Miguel Borges, 2021, review article Abstract: Plants influenced the evolution of plant-dwelling stink bugs’ systems underlying communication with chemical and substrate-borne vibratory signals. Plant volatiles provides cues that increase attractiveness or interfere with the probability of finding a mate in the field. Mechanical properties of herbaceous hosts and associated plants alter the frequency, amplitude, and temporal characteristics of stink bug species and sex-specific vibratory signals. The specificity of pheromone odor tuning has evolved through highly specific odorant receptors located within the receptor membrane. The narrow-band low-frequency characteristics of the signals produced by abdomen vibration and the frequency tuning of the highly sensitive subgenual organ vibration receptors match with filtering properties of the plants enabling optimized communication. A range of less sensitive mechanoreceptors, tuned to lower vibration frequencies, detect signals produced by other mechanisms used at less species-specific levels of communication in a plant environment. Whereas the encoding of frequency-intensity and temporal parameters of stink bug vibratory signals is relatively well investigated at low levels of processing in the ventral nerve cord, processing of this information and its integration with other modalities at higher neuronal levels still needs research attention.
Published in DiRROS: 05.08.2024; Views: 209; Downloads: 209
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32. Prilagajanje gospodarjenja z gozdovi podnebnim spremembam: zasnova in predlog strategijAndrej Bončina, 2024, review article Abstract: Podnebne spremembe se kažejo v postopnem spreminjanju povprečnih vrednosti podnebnih spremenljivk in večji pogostnosti ekstremnih vremenskih dogodkov. Podnebne spremembe vplivajo na razvoj gozdov. Ocena ranljivosti gozdov je izhodišče za gospodarjenje z gozdovi, prilagojeno podnebnim spremembam. S prilagojenim gospodarjenjem izboljšamo odpornost in prožnost gozdov ter pospešimo tranzicijo gozdov. Prilagojeno gospodarjenje je opredeljeno s strategijami, smernicami in ukrepi. V prispevku predstavljamo i) zasnovo prilagojenega gospodarjenja z gozdovi, ii) predlog strategij za prilagajanje gozdov podnebnim spremembam v Sloveniji in iii) postopek vključevanja prilagoditvenega gospodarjenja v celovito zasnovo gospodarjenja z gozdovi.
Keywords: ocena ranljivosti, odpornost gozdov, prožnost gozdov, prilagajanje gospodarjenja, strategije, ukrepi, načrtovalni postopek
Published in DiRROS: 05.08.2024; Views: 188; Downloads: 53
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33. Reproductive biology, mating behavior, and vibratory communication of the brown-winged stink bug, Edessa meditabunda (Fabr.) (Heteroptera: Pentatomidae)Cleonor Cavalcante A. Silva, Raúl Alberto Laumann, Jonatas Barbosa Cavalcante Ferreira, Maria Carolina Blassioli Moraes, Miguel Borges, Andrej Čokl, 2012, original scientific article Abstract: We describe different aspects of the reproductive biology, mating behavior, and vibratory communication of the pentatomid Edessa meditabunda (Fabr.). This species shows lower copulation frequency and reproductive potential with longer sexual maturation period compared to other species of pentatomids. Females with multiple mating show increased fecundity when compared with single-mated females and both increased fecundity and reduced longevity when compared with virgin females. Courtship and mating behavior and vibratory signals are typical and similar to what was observed in other species of pentatomids, except that males started the courtship. These results constitute the first paper on biology, behavior, and vibratory communication among species of the subfamily Edessinae.
Keywords: animal ecology, animal communication
Published in DiRROS: 05.08.2024; Views: 305; Downloads: 242
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34. Gene expression profiling of recombinant protein producing E. coli at suboptimal growth temperatureMitja Mahnič, Špela Baebler, Andrej Blejec, Špela Jalen, Kristina Gruden, Viktor Menart, Simona Jevševar, 2012, original scientific article Abstract: Recent studies have revealed that at lower cultivation temperatures (25 °C) much higher percentage of correctly folded recombinant hG-CSF protein can be extracted from inclusion bodies. Hence, the goal of our research was to investigate mechanisms determining characteristics of non-classical inclusion bodies production using gene expression profiling, focusing on proteases and chaperones gene expression. Statistical analysis of microarray data showed prominent changes in energy metabolism, in metabolism of amino acids and nucleotides, as well as in biosynthesis of cofactors and secondary metabolites if the culture was grown below its optimal temperature. Moreover, 24 differentially expressed up to now known genes classified among proteases, chaperones and other heat or stress related genes. Among chaperones UspE and among proteases YaeL and YeaZ might play an important role in accumulation of correctly folded recombinant proteins. Membrane localized protease yaeL gene was found to have higher activity at 25 °C and is thus potentially functionally related to the more efficient recombinant protein production at lower temperatures. The results of this study represent advance in the understanding of recombinant protein production in E. coli. Genes potentially influencing production of recombinant protein at lower growth temperature represent basis for further research towards improvement of E. coli production strains as well as fermentation process.
Keywords: recombinant protein production, non-classical inclusion bodies, expression microarrays, YaeL protease, GroEL chaperone
Published in DiRROS: 05.08.2024; Views: 334; Downloads: 284
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35. Identification of plasma biomarker candidates in glioblastoma using an antibody-array-based proteomic approachKlemen Zupančič, Marjan Koršič, Urška Verbovšek, Primož Rožman, Tamara Lah Turnšek, Andrej Blejec, Kristina Gruden, Helena Motaln, Miomir Knežević, Matija Veber, Ana Herman, 2014, original scientific article Abstract: Background. Glioblastoma multiforme (GBM) is a brain tumour with a very high patient mortality rate, with a median survival of 47 weeks. This might be improved by the identification of novel diagnostic, prognostic and predictive
therapy-response biomarkers, preferentially through the monitoring of the patient blood. The aim of this study was to define the impact of GBM in terms of alterations of the plasma protein levels in these patients.
Materials and methods. We used a commercially available antibody array that includes 656 antibodies to analyse
blood plasma samples from 17 healthy volunteers in comparison with 17 blood plasma samples from patients with
GBM.
Results. We identified 11 plasma proteins that are statistically most strongly associated with the presence of GBM.
These proteins belong to three functional signalling pathways: T-cell signalling and immune responses; cell adhesion and migration; and cell-cycle control and apoptosis. Thus, we can consider this identified set of proteins as potential diagnostic biomarker candidates for GBM. In addition, a set of 16 plasma proteins were significantly associated with the overall survival of these patients with GBM. Guanine nucleotide binding protein alpha (GNAO1) was associated with both GBM presence and survival of patients with GBM.
Conclusions. Antibody array analysis represents a useful tool for the screening of plasma samples for potential cancer biomarker candidates in small-scale exploratory experiments; however, clinical validation of these candidates requires their further evaluation in a larger study on an independent cohort of patients.
Keywords: glioblastoma, proteomics, biomarker
Published in DiRROS: 02.08.2024; Views: 214; Downloads: 161
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36. Reverse transcriptase droplet digital PCR shows high resilience to PCR inhibitors from plant, soil and water samplesNejc Rački, Tanja Dreo, Ion Gutiérrez-Aguirre, Andrej Blejec, Maja Ravnikar, 2014, original scientific article Abstract: Background
Detection and quantification of plant pathogens in the presence of inhibitory substances can be a challenge especially with plant and environmental samples. Real-time quantitative PCR has enabled high-throughput detection and quantification of pathogens; however, its quantitative use is linked to standardized reference materials, and its sensitivity to inhibitors can lead to lower quantification accuracy. Droplet digital PCR has been proposed as a method to overcome these drawbacks. Its absolute quantification does not rely on standards and its tolerance to inhibitors has been demonstrated mostly in clinical samples. Such features would be of great use in agricultural and environmental fields, therefore our study compared the performance of droplet digital PCR method when challenged with inhibitors common to plant and environmental samples and compared it with quantitative PCR.
Results
Transfer of an existing Pepper mild mottle virus assay from reverse transcription real-time quantitative PCR to reverse transcription droplet digital PCR was straight forward. When challenged with complex matrices (seeds, plants, soil, wastewater) and selected purified inhibitors droplet digital PCR showed higher resilience to inhibition for the quantification of an RNA virus (Pepper mild mottle virus), compared to reverse transcription real-time quantitative PCR.
Conclusions
This study confirms the improved detection and quantification of the PMMoV RT-ddPCR in the presence of inhibitors that are commonly found in samples of seeds, plant material, soil, and wastewater. Together with absolute quantification, independent of standard reference materials, this makes droplet digital PCR a valuable tool for detection and quantification of pathogens in inhibition prone samples.
Keywords: PCR amplification, inhibition, qPCR, droplet digital PCR, environmental samples
Published in DiRROS: 02.08.2024; Views: 210; Downloads: 120
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37. Evaluation of cyanobacteria biomass derived from upgrade of phycocyanin fluorescence estimationTinkara Rozina, Bojan Sedmak, Maja Zupančič Justin, Andrej Meglič, 2017, original scientific article Abstract: The number of harmful cyanobacterial blooms has increased significantly at the global level in recent years. One of the characteristics of cyanobacteria that gives them advantage over other phytoplankton organisms are auxiliary photosynthetic pigments, such as phycocyanin. This fluorescent pigment emits light at a different wavelength as chlorophyll and can therefore be used for detection of cyanobacteria in situ. In this study we used submersible phycocyanin fluorescence sensors and compare their voltage output to concentration of extracted phycocyanin, cell counts and biovolume. The relation was linear in all three cases; however, the variability of regression line slopes between different cyanobacteria strains was high in the case of PC extract concentration and cell count. The highest uniformity in the linear fits was between fluorescence signal and biovolume therefore making it the best candidate for fluorescence sensor voltage output conversion. In the context of this work we also compared different methods for PC extraction. Modifying the equations by subtracting the absorption at 750 nm almost entirely reduces the false PC concentration estimation due to sample turbidity.
Keywords: fluorescence measurements, phycocyanin, cyanobacteria
Published in DiRROS: 01.08.2024; Views: 212; Downloads: 102
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38. Combined use of chlorophyll a and phycocyanin fluorescence sensors for quantification and differentiation of phytoplankton : a useful approach for small surface water bodiesTinkara Rozina, Tina Eleršek, Maja Zupančič Justin, Andrej Meglič, 2018, original scientific article Abstract: Sensors based on in vivo measurements of photosynthetic pigments fluorescence enable real-time phytoplankton monitoring with high spatial and temporal resolution. A combination of chlorophyll a (CHL) and phycocyanin (PC) fluorescence sensors was used for phytoplankton quantification and differentiation in two small water bodies, Koseze Pond and pond in Hotinja vas. The high correlation of CHL and PC fluorescence signals with biovolume was confirmed during the two-year monitoring in anatural pond environment in spite of a seasonal succession of the phytoplankton. Additionally, disturbances of the sensors were investigated. Water bodies containing predominantly algae yielded false positive signals of the PC sensor, which reached up to 1% of the intensity of the CHL signal. Similarly, underestimated counts of cyanobacteria measured with CHL fluorescence sensor can be adjusted using PC fluorescence sensor.
Keywords: small water bodies, algae, cyanobacteria, fluorescence sensors, biovolume
Published in DiRROS: 01.08.2024; Views: 187; Downloads: 130
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39. Transcriptome study and identification of potential marker genes related to the stable expression of recombinant proteins in CHO clonesAndrej Blejec, Marjanca Blas, Petra Nikolić, Dominik Gaser, Kristina Gruden, Aleš Belič, Špela Baebler, Uroš Jamnikar, Andrej Francky, Holger Laux, 2015, original scientific article 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: 222; Downloads: 279
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40. The effect of timing of female vibrational reply on male signalling and searching behaviour in the leafhopper Aphrodes makaroviMeta Virant-Doberlet, Maarten De Groot, Andrej Blejec, Ana Kuhelj, 2015, original scientific article Abstract: Sexual communication in animals often involves duetting characterized by a coordinated reciprocal exchange of acoustic signals. We used playback experiments to study the role of timing of a female reply in the species-specific duet structure in the leafhopper Aphrodes makarovi (Hemiptera: Cicadellidae). In leafhoppers, mate recognition and location is mediated exclusively by species- and sex-specific substrate-borne vibrational signals and a female signal emitted in reply to male advertisement calls is essential for recognition and successful location of the female. In A. makarovi, males have to initiate each exchange of vibrational signals between partners, and in a duet the beginning of a female reply overlaps the end of the male advertisement call. Results of playback treatments in which female replies were delayed and did not overlap with the male call revealed that in order to trigger an appropriate behavioural response of the male, female reply has to appear in a period less than 400 ms after the end of the initiating male call. Results also suggest that males are not able to detect a female reply while calling, since female reply that did not continue after the end of male call triggered male behaviour similar to behaviour observed in the absence of female reply. Together, our results show that vibrational duets are tightly coordinated and that the species-specific duet structure plays an important role in mate recognition in location processes.
Keywords: vibration, insects, bioacoustics, animal signaling and communication
Published in DiRROS: 29.07.2024; Views: 193; Downloads: 143
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