NEFROPATIJA UZROKOVANA BK VIRUSOM U BOLESNIKA S TRANSPLANTIRANIM SRCEM: PRVI DOKUMENTIRANI SLUČAJ U HRVATSKOJ

As outcomes following heart transplantation have improved significantly over the last years, chronic kidney disease has become an increasingly prevalent complication in this population. Polyomavirus-associated nephropathy (PVAN) of native kidneys has also been recognized increasingly as a cause of kidney failure. We report the first case of PVAN occurring in the native kidneys of a solid-organ transplant recipient in Croatia as the eighth case described in the literature worldwide. A 65-year-old female with dilatative cardiomyopathy and good kidney function had a heart transplanted in 2012. Initial immunosuppressive therapy consisted of antithymocyte immunoglobulin with cyclosporine, mycophenolate mofetil and corticosteroids. Soon after transplantation, her kidney function began to fail progressively. Biopsy of the native kidneys revealed PVAN, and everolimus was introduced in immunosuppressive therapy. Nevertheless, her renal dysfunction progressed and she is now being evaluated for cadaveric kidney transplantation. PVAN should be considered in the differential diagnosis of new-onset renal failure following non-kidney solid organ transplantation. Early diagnosis is essential for prevention of irreversible renal damage.Unazad nekoliko godina sa značajnim poboljšanjem preživljenja bolesnika s transplantiranim srcem kronično zatajenje bubrega postalo je sve češća komplikacija u toj populaciji. Nefropatija uzrokovana poliomavirusom (PVAN) nativnih bubrega također se sve češće prepoznaje kao uzrok zatajenja bubrega. Prikazujemo prvi slučaj PVAN nativnih bubrega kod primatelja transplantata solidnog organa u Hrvatskoj i osmi takav slučaj dosad opisan u literaturi. Bolesnici u dobi od 65 godina s dilatativnom kardiomiopatijom i dobrom bubrežnom funkcijom je 2012. godine transplantirano srce. Inicijalna imunosupresivna terapija sastojala se od antitimocitnog imunoglobulina s ciklosporinom, mikofenolat mofetilom i kortikosteroidima. Ubrzo nakon transplantacije dolazi do zatajenja bubrega. Biopsijom nativnih bubrega postavljena je dijagnoza PVAN, a u imunosupresivnu terapiju je uveden everolimus. Usprkos tome dolazi do daljnjeg napredovanja zatajenja bubrega i bolesnica je trenutno u pripremi za kadaveričnu transplantaciju bubrega. PVAN treba razmotriti u diferencijalnoj dijagnozi novonastalog zatajenja bubrega nakon transplantacije solidnih organa. Rana dijagnoza PVAN je bitna u sprječavanju razvoja ireverzibilnog bubrežnog zatajenja

Genetic diversity of Nepeta rtanjensis based on RAPD data

Nepeta rtanjensis Diklić et Milojević (Lamiaceae) is an endemic, herbaceous plant, found only in a very small area of Mount Rtanj in Southeast Serbia. The whole species is represented by several hundred specimens distributed within three known populations: Greda, Mirčin del and Javor. Like many species of the genus Nepeta, N. rtanjensis is also famous for its secondary metabolite production (mainly nepetalactones and phenolics) and it is well known that these compounds show antibacterial, antifungal, cytotoxic, phytotoxic and antioxidant activities. Because of its limited distribution and the status of critically endangered species, it is important to find as much as possible information about the population genetic structure of this species. Therefore, we performed randomly amplified polymorphic DNA (RAPD) analysis to get an insight into the genetic diversity and variation of this species within and between the populations. DNA material used in this process was isolated from leaves collected from plants in their natural habitats. To our knowledge, this is the first employment of RAPD markers in the assessment of the N. rtanjenis genetic structure. Genetic variability between populations was not detected, but only small variations between individual plants from the same populations were recorded. These preliminary data give us a good base for further studies towards elucidating the genetic structure of this rare species using more reliable EST-SSR markers, designed based on N. rtanjensis transcriptome

Chemosensory Responses to the Repellent Nepeta Essential Oil and Its Major Component Nepetalactone by Aedes aegypti (Diptera: Culicidae), a Vector of Zika Virus

Nepeta essential oil (Neo; catnip) and its major component, nepetalactone, have long been known to repel insects including mosquitoes. However, the neural mechanisms through which these repellents are detected by mosquitoes, including the yellow fever mosquito Aedes aegypti (L.), an important vector of Zika virus, were poorly understood. Here we show that Neo volatiles activate olfactory receptor neurons within the basiconic sensilla on the maxillary palps of female Ae. aegypti. A gustatory receptor neuron sensitive to the feeding deterrent quinine and housed within sensilla on the labella of females was activated by both Neo and nepetalactone. Activity of a second gustatory receptor neuron sensitive to the feeding stimulant sucrose was suppressed by both repellents. Our results provide neural pathways for the reported spatial repellency and feeding deterrence of these repell ents. A better understanding of the neural input through which female mosquitoes make decisions to feed will facilitate design of new repellents and management strategies involving their use.The final publication is now available at: [https://doi.org/10.1093/jme/tjx059

Pursuit for est microsatellites in a tetraploid model from de novo transcriptome sequencing

Available scientific literature reports very few microsatellite markers derived from tetraploid genomes using de novo transcriptome sequencing, mostly because their gain usually represents a major computational challenge due to complicated combinatorics during assembly of sequence reads. Here we present a novel approach for mining polymorphic microsatellite loci from transcriptome data in a tetraploid species with no reference genome available. Pairs of 114 bp long de novo sequenced transcriptome reads of Centaurium erythraea were merged into short contigs of 170-200 bp each. High accuracy assembly of the pairs of reads was accomplished by a minimum of 14 bp overlap. Sequential bioinformatics operations involved fully free and open-source software and were performed using an average personal computer. Out of the 13 150 candidate contigs harboring SSR motifs obtained in a final output, we randomly chose 16 putative markers for which we designed primers. We tested the effectiveness of the established bioinformatics approach by amplifying them in eight different taxa within the genus Centaurium having various ploidy levels (diploids, tetraploids and hexaploids). Nine markers displayed polymorphism and/or transferability among studied taxa. They provided 54 alleles in total, ranging from 2 to 14 alleles per locus. The highest number of alleles was observed in C. erythraea, C. littorale and a hybridogenic taxon C. pannonicum. The developed markers are qualified to be used in genetic population studies on declining natural populations of Centaurium species, thus providing valuable information to evolutionary and conservation biologists. The developed cost-effective methodology provides abundant de novo assembled short contigs and holds great promise to mine numerous additional EST-SSR-containing markers for possible use in genetics population studies of tetraploid taxa within the genus Centauriu

Phenotypic and Genetic Variation of an Interspecific Centaurium Hybrid (Gentianaceae) and Its Parental Species.

Interspecific hybridization is one of the major actuators of evolutionary changes in plants. As the result of allopolyploid hybridization, offspring may gain different ploidy levels in comparison to parental species, which can provide them instant reproductive isolation. Two tetraploid sister species, Centaurium erythraea and C. littorale, readily cross-fertilize, resulting in hybrids of various ploidy. In northern Serbia, two stable populations of a hexaploid taxon C. pannonicum have been documented. It has been proposed previously that this taxon emerged after an interspecific hybridization event between two tetraploid sister-species: C. erythraea and C. littorale subsp. compressum. The existing populations of the hybridogenic taxon, as well as neighboring populations of the two parental taxa were here characterized by both morphometrics and molecular markers (EST-SSR and trnL-F). Three leaf and two flower characteristics were found to be informative in delimitation of the parental taxa and in their discernment from hybrid individuals, the latter having intermediate values. Eight microsatellite markers were found to have good ability to distinguish studied taxa, placing C. pannonicum in closer relationship with C. erythraea. Conversely, trnL-F plastid marker nominated C. littorale subsp. compressum to be the donor of the C. pannonicum plastid DNA. Reproductive isolation of the hexaploid hybrid individuals from the parental species should be examined as the next logical step in describing the new species

Functional iridoid synthases from iridoid producing and non-producing Nepeta species (subfam. Nepetoidae, fam. Lamiaceae)

Iridoids, a class of atypical monoterpenes, exhibit exceptional diversity within the Nepeta genus (subfam. Nepetoidae, fam. Lamiaceae).The majority of these plants produce iridoids of the unique stereochemistry, with nepetalactones (NLs) predominating; however, a few Nepeta species lack these compounds. By comparatively analyzing metabolomics, transcriptomics, gene co-expression, and phylogenetic data of the iridoid-producing N. rtanjensis Diklić & Milojević and iridoid-lacking N. nervosa Royle & Bentham, we presumed that one of the factors responsible for the absence of these compounds in N. nervosa is iridoid synthase (ISY). Two orthologues of ISY were mined from leaves transcriptome of N. rtanjensis (NrPRISE1 and NrPRISE2), while in N. nervosa only one (NnPRISE) was identified, and it was phylogenetically closer to the representatives of the Family 1 isoforms, designated as P5βRs. Organ-specific and MeJA-elicited profiling of iridoid content and co-expression analysis of IBG candidates, highlighted NrPRISE2 and NnPRISE as promising candidates for ISY orthologues, and their function was confirmed using in vitro assays with recombinant proteins, after heterologous expression of recombinant proteins in E. coli and their His-tag affinity purification. NrPRISE2 demonstrated ISY activity both in vitro and likely in planta, which was supported by the 3D modeling and molecular docking analysis, thus reclassification of NrPRISE2 to NrISY is accordingly recommended. NnPRISE also displays in vitro ISY-like activity, while its role under in vivo conditions was not here unambiguously confirmed. Most probably under in vivo conditions the NnPRISE lacks substrates to act upon, as a result of the loss of function of some of the upstream enzymes of the iridoid pathway. Our ongoing work is conducted towards re-establishing the biosynthesis of iridoids in N. nervosa

Supplementary data for the article: Aničić, N.; Gašić, U.; Lu, F.; Ćirić, A.; Ivanov, M.; Jevtić, B.; Dimitrijević, M.; Anđelković, B.; Skorić, M.; Nestorović Živković, J.; Mao, Y.; Liu, J.; Tang, C.; Soković, M.; Ye, Y.; Mišić, D. Antimicrobial and Immunomodulating Activities of Two Endemic Nepeta Species and Their Major Iridoids Isolated from Natural Sources. Pharmaceuticals 2021, 14 (5), 414. https://doi.org/10.3390/ph14050414.

Supplementary material for: [https://doi.org/10.3390/ph14050414]Related to published version: [https://cherry.chem.bg.ac.rs/handle/123456789/4444

Phytochemical characterization and antioxidant potential of rustyback fern (Asplenium ceterach L.)

Asplenium ceterach (syn. Ceterach officinarum Willd.) or rustyback fern, is a plant species traditionally used in Mediterranean countries as expectorant, diuretic, against spleen complaints, kidney stones and hemorrhoids. Phytochemical analysis of gametophytes and sporophytes of A. ceterach was performed, followed by comparative analysis of phenolic composition and the antioxidant properties of the extracts (scavenging capacities against ABTS·+ and DPPH·).Totally 16 phenolic compounds belonging to the classes of phenolic acids (hydroxybenzoic and hydroxycynnamic acids), flavonoids (flavan-3-ols, flavonols), and xanthones were identified using UHPLC/DAD/–HESI-MS/MS analysis. Differential phenolics’ composition of the two phases of the life cycle of this fern was observed which significantly determined their radical scavenging activities. In sporophytes of A. ceterach considerably high amounts of chlorogenic acid were found, while xanthones were mostly abundant in gametophytes, recommending this fern species as a valuable source of bioactive compounds

Diploid vs. tetraploid Centaurium erythraea Rafn: a comparative study of regenerative in vitro potential and biosynthetic capacity

The impact of ploidy level on both the regenerative potential under in vitro conditions and the production of major bioactive specialized metabolites, such are iridoids and xanthones, was examined in Centaurium erythraea Rafn. Shoot regeneration frequency was genotype dependent, but not affected by explant ploidy level. In most cases, the regenerated shoots of autotetraploid (4x) genotypes were more robust than diploid (2x) ones. Regeneration efficiency of root explants declined from the apical to the basal root segment. Shoot and root biomass production of two month-old shoots was not significantly different between 2x and 4x genotypes. Both 4x and 2x genotypes were characterized by the predominance of secoiridoid glucoside gentiopicrin in shoots and roots, which is followed by sweroside and swertiamarin. Loganic acid, loganin and secologanin were much less abundant. Methylbellidifolin was the major xanthone in both shoots and roots. Diploid plants showed higher biosynthetic capacity for the production of secoiridoids and xanthones in both shoots and roots. Results highlight a higher potential of diploid C. erythraea genotypes for biotechnology-based sustainable production of secoiridoids in comparison to tetraploid genotypes