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Parent-Offspring Relationships Following Mass Spawning of Wild Adriatic Sea Bass (Dicentrarchus labrax)

Jelena Lončar1, Renata Barić2, Lav Bavčević3 and Branko Kozulić1*


1
Gentius d.o.o, Petra Kasandrića 6, HR-23000 Zadar, Croatia

2Cromaris d.d., Gaženička cesta 4b, HR-23000 Zadar, Croatia
3Croatian Agricultural Advisory Service, Ivana Mažuranića 30, HR-23000 Zadar, Croatia

Article history:

Received May 14, 2013

Accepted: December 18, 2013

Key words:

sea bass, selective breeding, genotyping, microsatellites

Summary:

We have genotyped 44 fishes caught in the wild (Novigrad Sea, Croatia) using 11
microsatellite tetranucleotide markers. They were divided in two groups and after natural mass spawning, we selected 500 offspring for genotyping following their labelling withn electronic tags. All fish in the parental group could be identified based on unique genotypes, and a significant number of private alleles, the alleles found only in one fish, greatly facilitated subsequent parent-offspring assignments. The majority of potential parents left no progeny, while just six of them generated over 70 %. Such highly skewed reproduction success, observed also in other studies, can quickly lead to a highly inbred population in just a few selection cycles. This kind of genetic analysis is helpful for planning of future selective breeding, where faster progress will be made possible based on the soon to be completed sea bass genome sequence.

 

 

*Corresponding author:        branko.kozulic@yahoo.com
                                                          +385 23 331 077

                                            +385 23 331 08

 


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Flavonoids as Inhibitors of Human Butyrylcholinesterase Variants

Maja Katalinić§, Anita Bosak§ and Zrinka Kovarik*


Institute for Medical Research and Occupational Health, Ksaverska cesta 2, HR-10000 Zagreb, Croatia

Article history
:

Received March 25, 2013
Accepted September 23, 2013

Key words:

cholinesterase, reversible inhibition, galangin, quercetin, fisetin, luteolin

Summary:

The inhibition of butyrylcholinesterase (BChE, EC 3.1.1.8) appears to be of interest in
treating diseases with symptoms of reduced neurotransmitter levels, such as Alzheimer’s disease. However, BCHE gene polymorphism should not be neglected in research since it could have an effect on the expected outcome. Several well-known cholinergic drugs (e.g. galantamine, huperzine and rivastigmine) originating from plants, or synthesised as derivatives of plant compounds, have shown that herbs could serve as a source of novel target-directed compounds. We focused our research on flavonoids, biologically active polyphenolic compounds found in many plants and plant-derived products, as BChE inhibitors. All of the tested flavonoids: galangin, quercetin, fisetin and luteolin reversibly inhibited usual, atypical, and fluoride-resistant variants of human BChE. The inhibition potency increased in the following order, identically for all three BChE variants: luteolin<fisetin< quercetin<galangin. The determined enzyme-inhibitor dissociation constants (Ki) ranged from 10 to 170 μmol/L. We showed that no significant change in the inhibition potency of selected flavonoids exists in view of BChE polymorphism. Our results suggested that flavonoids could assist the further development of new BChE-targeted drugs for treating symptoms of neurodegenerative diseases and dementia.

 

 

*Corresponding author:        zkovarik@imi.hr                                        
                                       
   +385 1 4682 555
                                            +385 1 4673 303

§Both authors contributed equally to this work


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Ion-Exchange Sample Displacement Chromatography as a Method for Fast and Simple Isolation of Low- and High-Abundance Proteins from Complex Biological Mixtures

Martina Srajer Gajdosik1, Spomenka Kovac1, Nela Malatesti2, Egbert Müller3
and Djuro Josic2,4*

1
Department of Chemistry, Josip Juraj Strossmayer University, Cara Hadrijana 8/A,
HR-31000 Osijek, Croatia

2Department of Biotechnology, University of Rijeka, Radmile Matejčić 2, HR-51000 Rijeka, Croatia
3Tosoh Bioscience GmbH, Zettachring 6, DE-70567 Stuttgart, Germany
4Warren Alpert Medical School, Brown University, Richmond Street 222, RI-02903 Providence, USA

Article history:

Received April 18, 2013

Accepted December 18, 2013

Key words:

sample displacement chromatography, ion-exchange mode, plasma proteins

Summary:

Sample displacement chromatography (SDC) in reversed phase and ion-exchange
modes was introduced at the end of 1980s. This chromatographic method was first used for preparative purification of synthetic peptides, and subsequently adapted for protein fractionation, mainly in anion-exchange mode. In the past few years, SDC has been successfully used for enrichment of low- and medium-abundance proteins from complex biological fluids on both monolithic and bulk chromatographic supports. If aqueous mobile phase is used with the application of mild chromatographic conditions, isolated proteins are not denatured and can also keep their biological activity. In this paper, the use of SDC in anion-exchange mode on a high-capacity chromatographic resin for separation of proteins from complex biological mixtures such as human plasma is demonstrated. By use of three and more columns coupled in series during sample application, and subsequent parallel elution of detached columns, additional separation of bound proteins was achieved. Highly enriched human serum albumin fraction and a number of physiologically active medium- and low-abundance proteins could be fractionated and detected by electrospray ionization tandem mass spectrometry (ESI-MS/MS) and matrix-assisted laser desorption/ ionization time-of-flight tandem mass spectrometry (MALDI-TOF/TOF-MS). The use of the aforementioned columns that can be sanitized with 1 M sodium hydroxide for further application of SDC in biotechnology and food technology was discussed.


 

*Corresponding author:        djuro_josic@brown.edu                                        
                                       
   +385 51 584 560
                                            +385 51 584 599

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Generation of New Genotypic and Phenotypic Features in Artificial and Natural Yeast Hybrids

Walter P. Pfliegler1, Lea Atanasova2, Edina Karanyicz1, Matthias Sipiczki1,
Ursula Bond3, Irina S. Druzhinina2, Katja Sterflinger4 and Ksenija Lopandic4*


1
Department of Genetics and Applied Microbiology, University of Debrecen, Egyetem tér 1,
H-4032 Debrecen, Hungary

2
Research Area Biotechnology and Microbiology, Institute of Chemical Engineering,
Vienna University of Technology, Gumpendorferstrasse 1a, A-1060 Vienna, Austria

3
The School of Genetics and Microbiology, Trinity College, College Green, Dublin 2, Ireland

4VIBT-BOKU, University of Natural Resources and Applied Life Sciences,
Department of Biotechnology, Muthgasse 11, A-1190 Vienna, Austria


Article history:

Received March 29, 2013

Accepted September 23, 2013


Key words:

Saccharomyces cerevisiae, S. uvarum, S. kudriavzevii, yeast interspecies hybrids, AFLP, karyotyping


Summary:

Evolution and genome stabilization have mostly been studied on the Saccharomyces hybrids isolated from natural and alcoholic fermentation environments. Genetic and phenotypic properties have usually been compared to the laboratory and reference strains, as the true ancestors of the natural hybrid yeasts are unknown. In this way the exact impact of different parental fractions on the genome organization or metabolic activity of the hybrid yeasts is difficult to resolve completely. In the present work the evolution of geno- and phenotypic properties is studied in the interspecies hybrids created by the cross-breeding of S. cerevisiae with S. uvarum or S. kudriavzevii auxotrophic mutants. We hypothesized that the extent of genomic alterations in S. cerevisiae × S. uvarum and S. cerevisiae × S. kudriavzevii should affect the physiology of their F1 offspring in different ways. Our results, obtained by amplified fragment length polymorphism (AFLP) genotyping and karyotyping analyses, showed that both subgenomes of the S. cerevisiae x S. uvarum and of S. cerevisiae × S. kudriavzevii hybrids experienced various modifications. However, the S. cerevisiae × S. kudriavzevii F1 hybrids underwent more severe genomic alterations than the S. cerevisiae × S. uvarum ones. Generation of the new genotypes also influenced the physiological performances of the hybrids and the occurrence of novel phenotypes. Significant differences in carbohydrate utilization and distinct growth dynamics at increasing concentrations of sodium chloride, urea and miconazole were observed within and between the S. cerevisiae × S. uvarum and S. cerevisiae × S. kudriavzevii hybrids. Parental strains also demonstrated different contributions to the final metabolic outcomes of the hybrid yeasts. A comparison of the genotypic properties of the artificial hybrids with several hybrid isolates from the wine-related environments and wastewater demonstrated a greater genetic variability of the S. cerevisiae × S. kudriavzevii hybrids. Saccharomyces cerevisiae × S. uvarum artificial and natural hybrids showed considerable differences in osmolyte tolerance and sensitivity to miconazole, whereas the S. cerevisiae × S. kudriavzevii hybrids exhibited differences also in maltotriose utilization. The results of this study suggest that chromosomal rearrangements and genomic reorganizations as post-hybridization processes may affect the phenotypic properties of the hybrid progeny substantially. Relative to their ancestors, the F1 segregants may generate different phenotypes, indicating novel routes of evolution in response to environmental growth conditions.
 


*Corresponding author:
       
ksenija.lopandic@boku.ac.at                                         
                                      
   +43 1 47654 6943

                                          +43 1 47654 6675
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