Lea Vojta - Academia.edu (original) (raw)

Papers by Lea Vojta

Parasitology Research, 2017

The bacteria Anaplasma platys, Anaplasma phagocytophilum and Ehrlichia canis are tick-borne agent... more The bacteria Anaplasma platys, Anaplasma phagocytophilum and Ehrlichia canis are tick-borne agents that cause canine vector-borne disease. The prevalence of these pathogens in South Eastern Europe is unknown with the exception of an isolated case of A. platys detected in a dog imported into Germany from Croatia. To gain a better insight into their presence and prevalence, PCR-based screening for these bacterial pathogens was performed on domesticated dogs from different regions of Croatia. Blood samples from 1080 apparently healthy dogs from coastal and continental parts of Croatia as well as tissue samples collected from 63 deceased dogs with a history of anaemia and thrombocytopenia were collected for molecular screening by an Anaplasmataceaespecific 16S rRNA conventional PCR. Positive samples were confirmed using a second Anaplasmataceae-specific PCR assay with the PCR product sequenced for the purpose of bacterial species identification. All sequenced isolates were georeferenced and a kernel intensity estimator was used to identify clusters of greater case intensity. 42/1080 (3.8%; CI 2.7-5.0) of the healthy dogs were PCR positive for bacteria in the Anaplasmataceae. Sequencing of the 16S rRNA gene amplified from these positive samples revealed the presence of A. p l a t y s i n 2. 5 % (C I 1. 6-3. 4 % , 2 7 d o g s) , A. phagocytophilum in 0.3% (CI 0-0.6%, 3 dogs) and a Wolbachia endosymbiont in 1.1% (CI 0.4-1.6%, 12 dogs) of dogs screened in this study. Necropsied dogs were free from infection. Notably, no evidence of E. canis infection was found in any animal. This survey represents a rare molecular study of Anaplasmataceae in dogs in South Eastern Europe, confirming the presence of A. platys and A. phagocytophilum but not E. canis. The absence of E. canis was surprising given it has been described in all other Mediterranean countries surveyed and raises questions over the regional vector capacity of the Rhipicephalus sanguineus tick.

Frontiers in Plant Science, Mar 24, 2020

Periodicum biologorum, 2002

Background and Purpose: The molecular chaperones, peptidyl-prolyl isomerases (PPIases), catalyse ... more Background and Purpose: The molecular chaperones, peptidyl-prolyl isomerases (PPIases), catalyse isomerization around peptide bonds which neighbour proline residues. They participate in a wide variety of important cellular processes, although their true ...

Journal of Molecular Structure

Journal of Plant Physiology

International Journal of Molecular Sciences

Thylakoid rhodanase-like protein (TROL) is involved in the final step of photosynthetic electron ... more Thylakoid rhodanase-like protein (TROL) is involved in the final step of photosynthetic electron transport from ferredoxin to ferredoxin: NADP + oxidoreductase (FNR). TROL is located in two distinct chloroplast compartments-in the inner envelope of chloroplasts, in its precursor form; and in the thylakoid membranes, in its fully processed form. Its role in the inner envelope, as well as the determinants for its differential localization, have not been resolved yet. In this work we created six N-terminal amino acid substitutions surrounding the predicted processing site in the presequence of TROL in order to obtain a construct whose import is affected or localization limited to a single intrachloroplastic site. By using in vitro transcription and translation and subsequent protein import methods, we found that a single amino acid exchange in the presequence, Ala67 to Ile67 interferes with processing in the stroma and directs the whole pool of in vitro translated TROL to the inner envelope of chloroplasts. This result opens up the possibility of studying the role of TROL in the chloroplast inner envelope as well as possible consequence/s of its absence from the thylakoids.

Data in brief, 2016

In photosynthesis, the flavoenzyme ferredoxin:NADP(+) oxidoreductase (FNR) catalyses the final el... more In photosynthesis, the flavoenzyme ferredoxin:NADP(+) oxidoreductase (FNR) catalyses the final electron transfer from ferredoxin to NADP(+), which is considered as the main pathway of high-energy electron partitioning in chloroplasts (DOI: 10.1111/j.1365-313X.2009.03999.x[1], DOI: 10.1038/srep10085[2]). Different detergents and pH treatments of photosynthetic membranes isolated from the Arabidopsis wild-type (WT) and the loss-of-function mutants of the thylakoid rhodanase-like protein TROL (trol), pre-acclimated to either dark, growth-light, or high-light conditions, were used to probe the strength of FNR-membrane associations. Detergents β-DM (decyl-β-D-maltopyranoside) or β-DDM (n-dodecyl-β-D-maltopyranoside) were used to test the stability of FNR binding to the thylakoid membranes, and to assess different membrane domains containing FNR. Further, the extraction conditions mimicked pH status of chloroplast stroma during changing light regimes. Plants without TROL are incapable of ...

Import of a hybrid construct consisting of the transit sequence of SSU, the N-proximal part of ma... more Import of a hybrid construct consisting of the transit sequence of SSU, the N-proximal part of mature Tic110 and the mature SSU into chloroplasts led to the appearance of a soluble stromal import intermediate and the proposal that Tic110 might use a re-export pathway from the stroma to the inner envelope membrane. For full length Tic110 no soluble intermediate has been observed yet. One of the goals of this work was to investigate the import pathway of Tic110 in more detail. In this research the soluble stromal intermediate of Tic110 was observed, its re-export to the membrane was followed, and finally, the intermediate was isolated and co-immunoprecipitated with the stromal chaperones Hsp93, Hsp70 and to a lesser extent Cpn60. The obtained results indicate that Tic110, as proposed, uses a re-export pathway (conservative sorting) during its import into the chloroplast inner envelope membrane. Tic110 also requires stromal chaperones for achieving its native conformation, prior to the...

Periodicum Biologorum

Background and purpose: The molecular chaperones, peptidyl-prolyl isomerases (PPIases), catalyse ... more Background and purpose: The molecular chaperones, peptidyl-prolyl isomerases (PPIases), catalyse isomerization around peptide bonds which neighbour proline residues. They participate in a wide variety of important cellular processes, although their true physiological functions are still a subject of intense research. We recently identified a unique chloroplast PPIase, TLP40, which is involved in the regulation of oxygenic photosynthesis. Here, we describe the reverse genetic analysis of a cyanobacterial gene which could be the prokaryotic ancestor of TLP40 isomerase. Materials and Methods: The deduced amino acid sequence of the open reading frame sll0408 was analysed in detail and compared with related proteins. For the knock-out construct, the nucleotide sequence within the locus sll0408 was amplified using the polymerase chain reaction. The gentamycin-resistance cassette was inserted at a single BamHI site within the reading frame, and the construct was ligated with the pBluescrip...

Periodicum Biologorum, Mar 30, 2012

-dependent malic enzyme, which can form NADPH in an alternative pathway. Thus, NADPH synthesis, m... more -dependent malic enzyme, which can form NADPH in an alternative pathway. Thus, NADPH synthesis, mediated by FNR-TROL interaction, may be the source element in metabolic retrograde signal-transduction pathway linking light reactions with nuclear gene expression.

Veterinarski Arhiv

Infection with Hepatozoon canis has been described recently in 11.8% of Croatian dogs. Hematologi... more Infection with Hepatozoon canis has been described recently in 11.8% of Croatian dogs. Hematological and biochemical analyses of 14 apparently asymptomatic, but Hepatozoon-positive dogs were performed for the first time in Croatia. Haematological analyses showed severe eosinophilia in 9 cases (64.3%), leukocytosis in 5 (35.7%), neutrophilia in 3 (21.4%), decreased values of HCT and HGB in 2 (14.3%), monocytosis in 3 (21.4%), lymphocytosis in 6 (42.9%), thrombocytopenia in 2 (14.3%) and thrombocytosis in 3 cases (21.4%), where platelet aggregates were detected. Biochemical analyses of dog sera demonstrated highly increased ALP in 7 (50%) and ALT and CK in 2 dogs (14.3% each). AST and GGT were slightly increased in 2 (21.4%) samples. Also, β1-hyperglobulinemia, β1-hypoglobulinemia and γ-hypoglobulinemia were detected in certain samples (28.6%, 21.4% and 92.9%, respectively). Many changes in the biochemical and haematological parameters in this study correlate with previous researches,...

Biotechnology Reports, 2015

We report the production of hGM-CSF cytokine in leaves of industrial tobacco cultivars DH-17 and ... more We report the production of hGM-CSF cytokine in leaves of industrial tobacco cultivars DH-17 and DH-27 by using Agrobacterium-mediated transient expression. We prove the concept that very high biomass industrial tobacco plants are suitable platforms for rapid, low cost production of foreign proteins. Successful transient expression of the GM-CSF was achieved in less than three months, opening the possibility for future applications of this approach in rapid response production of various proteins of non-plant origin in industrial tobacco.

Veterinary Parasitology

An epizootiological field study on tick-borne protozoan infections in foxes (Vulpes vulpes) was c... more An epizootiological field study on tick-borne protozoan infections in foxes (Vulpes vulpes) was carried out in different parts of Croatia. Spleen samples of 191 carcasses of red foxes killed in sanitary hunting, were examined for the presence of hematozoa by polymerase chain reaction (PCR) and subsequent sequencing. The investigation revealed four species of hematozoa in 57 foxes (30%), namely Theileria annae, Theileria sp. 3182/05 and Hepatozoon canis. T. annae was found in 10 foxes (5%), Theileria sp. 3182/05 in a single animal (1%), H. canis in 44 (23%) and Hepatozoon sp. was detected in two foxes (1%). T. annae and H. canis were distributed through all the studied regions, while Theileria sp. 3182/05 and Hepatozoon sp. were restricted to the Zagreb and Zagorje, and Istria regions, respectively. Detection of T. annae in all regions of Croatia indicates the presence of the natural cycle of the parasite and raises the possibility of other vectors other than the proposed Ixodes hexa...

Scientific Reports, 2015

In photosynthesis, final electron transfer from ferredoxin to NADP + is accomplished by the flavo... more In photosynthesis, final electron transfer from ferredoxin to NADP + is accomplished by the flavo enzyme ferredoxin:NADP + oxidoreductase (FNR). FNR is recruited to thylakoid membranes via integral membrane thylakoid rhodanase-like protein TROL. We address the fate of electrons downstream of photosystem I when TROL is absent. We have employed electron paramagnetic resonance (EPR) spectroscopy to study free radical formation and electron partitioning in TROLdepleted chloroplasts. DMPO was used to detect superoxide anion (O 2 .− ) formation, while the generation of other free radicals was monitored by Tiron. Chloroplasts from trol plants pre-acclimated to different light conditions consistently exhibited diminished O 2 .− accumulation. Generation of other radical forms was elevated in trol chloroplasts in all tested conditions, except for the plants pre-acclimated to high-light. Remarkably, dark-and growth light-acclimated trol chloroplasts were resilient to O 2 .− generation induced by methyl-viologen. We propose that the dynamic binding and release of FNR from TROL can control the flow of photosynthetic electrons prior to activation of the pseudo-cyclic electron transfer pathway.

Advances in Photosynthesis - Fundamental Aspects, 2012

Veterinary Parasitology, 2010

The Plant Journal, 2000

Working in tandem, two photosystems in the chloroplast thylakoid membranes produce a linear elect... more Working in tandem, two photosystems in the chloroplast thylakoid membranes produce a linear electron flow from H 2 O to NADP + . Final electron transfer from ferredoxin to NADP + is accomplished by a flavoenzyme ferredoxin:NADP + oxidoreductase (FNR). Here we describe TROL (thylakoid rhodanese-like protein), a nuclearencoded component of thylakoid membranes that is required for tethering of FNR and sustaining efficient linear electron flow (LEF) in vascular plants. TROL consists of two distinct modules; a centrally positioned rhodanese-like domain and a C-terminal hydrophobic FNR binding region. Analysis of Arabidopsis mutant lines indicates that, in the absence of TROL, relative electron transport rates at high-light intensities are severely lowered accompanied with significant increase in non-photochemical quenching (NPQ). Thus, TROL might represent a missing thylakoid membrane docking site for a complex between FNR, ferredoxin and NADP + . Such association might be necessary for maintaining photosynthetic redox poise and enhancement of the NPQ.

FEBS Journal, 2007

The import of proteins destined for the intermembrane space of chloroplasts has not been investig... more The import of proteins destined for the intermembrane space of chloroplasts has not been investigated in detail up to now. By investigating energy requirements and time courses, as well as performing competition experiments, we show that the two intermembrane space components Tic22 and MGD1 (E.C. 2.4.1.46) both engage the Toc machinery for crossing the outer envelope, whereas their pathways diverge thereafter. Although MGD1 appears to at least partly cross the inner envelope, Tic22 very likely reaches its mature form in the intermembrane space without involving stromal components. Thus, different pathways for intermembrane space targeting probably exist in chloroplasts.