Search Results (17)

The main goal of this study is to present future changes in various precipitation indices at a kilometer-scale resolution for Bulgaria on an annual and seasonal basis. Numerical simulations were conducted using the Non-Hydrostatic Regional Climate Model version 4 (RegCM4-NH) following the Coordinated Regional Climate Downscaling Experiment Flagship Pilot Study protocol for three 10-year periods (1995–2004, 2041–2050, and 2090–2099), with horizontal grid resolutions of 15 km and 3 km, on the petascale supercomputer HPC Discoverer at Sofia Tech Park. Data from the Hadley Centre Global Environment Model version 2 (HadGEM2-ES), based on the Representative Concentration Pathway 8.5 (RCP8.5) scenario, were used as boundary conditions for the regional climate model (RCM) simulations, which were subsequently downscaled to the kilometer-scale (3 km) simulations using a one-way nesting approach. High-resolution model data were compared with high-resolution observational datasets as well as lower-resolution (15 km) data. Future changes in precipitation indices were analyzed on both annual and seasonal scales, including mean daily and hourly precipitation, the frequency and intensity of wet days (>1 mm/day) and wet hours (>0.1 mm/hour), extreme daily precipitation (99th percentile, p99), and extreme hourly precipitation (99.9th percentile, p99.9) for both future periods. Additionally, changes in near-surface (2 m) temperature and surface snow amount were also presented. There is no substantial difference in projected temperature change between the resolutions. A positive trend in annual mean precipitation is expected in the near future. Extreme precipitation (p99 and p99.9) is projected to increase in spring and winter, accompanied by a rise in daily and hourly precipitation intensity across both future periods. An increase in surface snow amount is observed in the central Danubian Plain, Thracian Lowland, and parts of the Rila and Pirin mountains for the near-future period. However, surface snow amount is expected to decrease by the end of the century. Full article

Background: Pirins are nuclear cupin proteins, one of several gene families within the plant cupin superfamily. However, the identification and functional analysis of Pirin proteins in Nicotiana benthamiana have not been explored. Methods: In this study, genome-wide analysis identifying NbPirin genes in N. benthamiana was conducted, as was phylogenetic analysis of Pirin genes in four Solanaceae species (including Capsicum annuum, Solanum lycopersicum, Solanum tuberosum, and N. benthamiana). In addition, we also evaluated the expression pattern of NbPirins under abiotic stress (temperature and phytohormones) and biotic stress (TMV, TuMV, and PVX). Results: A total of six Nbpirin genes were identified, which can be divided into three clades, and NbPirins also embraced a variety of abiotic or biotic cis-acting elements. The results showed that the expression of NbPirin1-6 was influenced by temperature variations, of which NbPirin6 was significantly upregulated at high temperatures (42 °C) but downregulated at low temperatures (4 °C). Notably, the expression of NbPirin6 exhibited a consistent decrease under ABA and MeJA treatments. Moreover, the expression of NbPirin1-6 was also affected by TMV, TuMV, and PVX infection. NbPirin1, NbPirin2, NbPirin3, and NbPirin5 showed higher expression levels under different viral infections compared to non-infection. Interestingly, NbPirin3 showed the highest expression level during TuMV infection (approximately a 20-fold increase compared to non-infection). Conclusions: Our study proposes the potential role of NbPirin6 in plant responses to abiotic stress, and the role of NbPirin3 in plant antiviral defense, and further lays the groundwork for future research on the functions of NbPirin proteins in responses to various stressors. Full article

In this study, we conducted a comprehensive survey aimed at assessing the diversity of yeast species inhabiting the guts of various insect species collected mainly from two Bulgarian National Parks, namely, Rila, and Pirin. The insect specimens encompass a broad taxonomic spectrum, including representatives from Coleoptera, Orthoptera, Lepidoptera, Hymenoptera, Dermaptera, Isopoda, and Collembola. Yeast strains were identified with DNA barcoding using the ribosomal markers, specifically, the D1/D2 domains of the ribosomal large subunit (LSU) and the internal transcribed spacers regions ITS 1 + 2 (ITS). The analysis unveiled the presence of 89 ascomycetous and 18 basidiomycetous yeast isolates associated with the insect specimens. Furthermore, our study identified 18 hitherto unknown yeast species. Full article

Extracellular vesicles (EVs) are functional substances secreted by microbes and host cells, and it has been discovered that they participate in the interactions between different microorganisms. Our recent findings indicate that Limosilactobacillus reuteri-derived EVs have the potential to improve the intestinal microbiota of Oplegnathus fasciatus fish and inhibit pathogenic bacteria. Previous research has reported that the host intestinal cells play a regulatory role in the intestinal microbiota. This suggested that to investigate the mechanisms through which L. reuteri-derived EVs regulate the intestinal microbiota, a system that excludes interference from host intestinal cells should be established. In this study, an in vitro cultured intestinal bacteria system, without host factors, was used to simulate the intestinal microbiota of O. fasciatus fish. After adding L. reuteri-derived EVs to the system, the changes in the microbiota were analyzed. The results showed that L. reuteri-derived EVs effectively reduced the abundance of Vibrio spp. In the results of the in vitro experiments, it was also observed that L. reuteri-derived EVs have the ability to inhibit Vibrio alginolyticus. We further sequenced the small RNA contained in L. reuteri-derived EVs and found that these small RNAs can interfere with genes (LysR, pirin, MIpA/OmpV, CatB, and aspartate-semialdehyde dehydrogenase) related to the growth of V. alginolyticus. Taken together, the results indicate that in the absence of host involvement, the small RNAs present in L. reuteri-derived EVs have the function of inhibiting pathogenic bacteria and exhibit the potential to regulate the intestinal microbiota. Full article

Candida albicans Prn1 is a protein with an unknown function similar to mammalian Pirin. It also has orthologues in other pathogenic fungi, but not in Saccharomyces cerevisiae. Prn1 highly increases its abundance in response to H₂O₂ treatment; thus, to study its involvement in the oxidative stress response, a C. albicans prn1∆ mutant and the corresponding wild-type strain SN250 have been studied. Under H₂O₂ treatment, Prn1 absence led to a higher level of reactive oxygen species (ROS) and a lower survival rate, with a higher percentage of death by apoptosis, confirming its relevant role in oxidative detoxication. The quantitative differential proteomics studies of both strains in the presence and absence of H₂O₂ indicated a lower increase in proteins with oxidoreductase activity after the treatment in the prn1∆ strain, as well as an increase in proteasome-activating proteins, corroborated by in vivo measurements of proteasome activity, with respect to the wild type. In addition, remarkable differences in the abundance of some transcription factors were observed between mutant and wild-type strains, e.g., Mnl1 or Nrg1, an Mnl1 antagonist. orf19.4850, a protein orthologue to S. cerevisiae Cub1, has shown its involvement in the response to H₂O₂ and in proteasome function when Prn1 is highly expressed in the wild type. Full article

The study describes a pilot survey on bats in the highest areas of Pirin Mountain. The methods included examining subfossil bone remains, mist-netting, and recording echolocation calls. The study was conducted in August 2002 and 2013 and from 2019 to 2020. While in general, bat diversity tends to decrease with increasing altitude due to harsher environmental conditions, the present study, despite a short period, reveals high diversity. Twenty species, more than half of the Bulgarian bat fauna, were detected. The recording and analysis of vocal signatures proved to be the best way to inventory bat diversity. At least 13 species were detected by this method. Vespertilio murinus and Tadarida teniotis together make up more than 60% of all reliably determined echolocation sequences. Significant activity was found for Myotis myotis/blythii, Plecotus auritus, Eptesicus serotinus, and E. nilssonii. The registration of the latter species is of considerable faunistic interest. It was previously only known from a single specimen at one location in the country. The sex and age structure of the bat assemblage suggests that it is likely a swarming assemblage. The area is the highest swarming location in Europe. The results provide valuable information on bat ecology and behaviour, which can be used to inform management and protection efforts. Full article

Acinetobacter baumannii is a nosocomial pathogen of serious healthcare concern that is becoming increasingly difficult to treat due to antibiotic treatment failure. Recent studies have revealed that clinically defined antibiotic-susceptible strains upregulate the expression of a repertoire of putative drug efflux pumps during their growth under biologically relevant conditions, e.g., in human serum, resulting in efflux-associated resistance to physiologically achievable antibiotic levels within a patient. This phenomenon, termed Adaptive Efflux Mediated Resistance (AEMR), has been hypothesized to account for one mechanism by which antibiotic-susceptible A. baumannii fails to respond to antibiotic treatment. In the current study, we sought to identify genetic determinants that contribute to A. baumannii serum-associated AEMR by screening a transposon mutant library for members that display a loss of the AEMR phenotype. Results revealed that mutation of a putative pirin-like protein, YhaK, results in a loss of AEMR, a phenotype that could be complemented by a wild-type copy of the yhaK gene and was verified in a second strain background. Ethidium bromide efflux assays confirmed that the loss of AEMR phenotype due to pirin-like protein mutation correlated with reduced overarching efflux capacity. Further, flow cytometry and confocal microscopy measures of a fluorophore 7-(dimethylamino)-coumarin-4-acetic acid (DMACA)-tagged levofloxacin isomer, ofloxacin, further verified that YhaK mutation reduces AEMR-mediated antibiotic efflux. RNA-sequencing studies revealed that YhaK may be required for the expression of multiple efflux-associated systems, including MATE and ABC families of efflux pumps. Collectively, the data indicate that the A. baumannii YhaK pirin-like protein plays a role in modulating the organism’s adaptive efflux-mediated resistance phenotype. Full article

In the Pirin Mountains, at an elevation of around 1000 m, three populations of a new species of Bulgarian flora from the genus Crocus, series Verni, were discovered. The species was compared to the morphologically related C. veluchensis, and presented with diagnostic morphological and anatomical features. Despite the high degree of morphological similarity, the molecular analysis, which included sequences from all related species (C. cvijicii, C. dalmaticus, C. jablanicensis, C. rujanensis, C. sieberi subsp. atticus, and C. veluchensis), distinguished the Pirin Mountains’ populations, and revealed the closest relationship to C. heuffelianus. Despite the C. heuffelianus/C. verni complex’s uncertain taxonomic status, our findings on the local population, based on morphometric, anatomical, molecular, and geographic analyses, indicate its belonging to the putative allotetraploid C. heuffelianus of south-eastern Europe and the Balkans, and an expansion of its range to the southeast. Given the taxonomic uncertainty and unclear phylogenetic relationships of the taxa in the Crocus vernus complex, we considered it appropriate to accept our taxon as Crocus heuffelianus. So far, only C. tommasinianus Herb. has been found in Bulgarian flora from the Crocus series Verni, but in terms of altitude and morphological features, the species from our collection is close to the Balkan endemic C. veluchensis, which belongs to the C. sieberi aggregate. Morphologically, it differs by the dark, heart-shaped spots on the tip of the tepals, and the presence of one bract. A detailed comparative anatomical analysis between the three species of crocuses from the series Verni in Bulgaria shows discrete differences: the width of the white stripe and lacunar area are good distinguishing features, as are the number of conducting vessels. Full article

This study set out to assess the performance of an artificial intelligence (AI) algorithm based on clinical data and dermatoscopic imaging for the early diagnosis of melanoma, and its capacity to define the metastatic progression of melanoma through serological and histopathological biomarkers, enabling dermatologists to make more informed decisions about patient management. Integrated analysis of demographic data, images of the skin lesions, and serum and histopathological markers were analyzed in a group of 196 patients with melanoma. The interleukins (ILs) IL-4, IL-6, IL-10, and IL-17A as well as IFNγ (interferon), GM-CSF (granulocyte and macrophage colony-stimulating factor), TGFβ (transforming growth factor), and the protein DCD (dermcidin) were quantified in the serum of melanoma patients at the time of diagnosis, and the expression of the RKIP, PIRIN, BCL2, BCL3, MITF, and ANXA5 proteins was detected by immunohistochemistry (IHC) in melanoma biopsies. An AI algorithm was used to improve the early diagnosis of melanoma and to predict the risk of metastasis and of disease-free survival. Two models were obtained to predict metastasis (including “all patients” or only patients “at early stages of melanoma”), and a series of attributes were seen to predict the progression of metastasis: Breslow thickness, infiltrating BCL-2 expressing lymphocytes, and IL-4 and IL-6 serum levels. Importantly, a decrease in serum GM-CSF seems to be a marker of poor prognosis in patients with early-stage melanomas. Full article

Methanosarcina spherical virus (MetSV), infecting Methanosarcina species, encodes 22 genes, but their role in the infection process in combination with host genes has remained unknown. To study the infection process in detail, infected and uninfected M. mazei cultures were compared using dual-RNAseq, qRT-PCRs, and transmission electron microscopy (TEM). The transcriptome analysis strongly indicates a combined role of virus and host genes in replication, virus assembly, and lysis. Thereby, 285 host and virus genes were significantly regulated. Within these 285 regulated genes, a network of the viral polymerase, MetSVORF6, MetSVORF5, MetSVORF2, and the host genes encoding NrdD, NrdG, a CDC48 family protein, and a SSB protein with a role in viral replication was postulated. Ultrastructural analysis at 180 min p.i. revealed many infected cells with virus particles randomly scattered throughout the cytoplasm or attached at the cell surface, and membrane fragments indicating cell lysis. Dual-RNAseq and qRT-PCR analyses suggested a multifactorial lysis reaction in potential connection to the regulation of a cysteine proteinase, a pirin-like protein and a HicB-solo protein. Our study’s results led to the first preliminary infection model of MetSV infecting M. mazei, summarizing the key infection steps as follows: replication, assembly, and host cell lysis. Full article

The Pirin Mountains in Bulgaria are refuge of endemic Pinus peuce and Pinus heldreichii forests. Due to the steep and long slopes, the forests are affected by avalanches and many trees keep record of the past avalanche activity in their tree rings. In our study, we used a combination of dendrochronology, satellite images and historical aerophotos to study the effects of avalanches on forests in the Bunderitsa valley. Our findings show that avalanches are the main shaping factor for the structure of forests in the valley, followed by fires. Past avalanche activity is responsible for the opened long-lasting avalanche tracks in the forests. About 60% of the potential forests (i.e., territories below the treeline, outside of the avalanche couloirs, streams, rock formations and screes) are strongly affected by avalanches. Of them, almost 40% are in avalanche runout zones, 12% are in avalanche tracks in the forests and 48% are forests, which are periodically strongly affected by bigger avalanches. Comparisons of newer satellite images with older aerophotos showed that in the 1970s, there were larger openings in the forests due to the high frequency of avalanches in the very snowy 1950s and 1960s. Although, recently, the avalanche activity has decreased, there are still years with larger avalanches, which affect forests. In addition, tourist development in the vicinity has increased the risk for human health and life due to avalanche accidents, including in forests. Tree-ring analysis allowed for the reconstructing of past avalanches that affected certain areas. The big couloirs are affected by smaller avalanches almost annually, while bigger avalanches have hit the neighboring forests almost every decade. Our findings demonstrate that avalanches in the valley are of a high importance and require more attention from authorities both as a risk factor for human health and life and as a natural disturbance, shaping the structure and dynamics of the forest. Full article

(1) Background: What is the role of serum CMV PCR in the diagnosis of recent primary infection (PI) in pregnant women when IgG avidity is uninformative? (2) Methods: Retrospective cohort study to compare serum versus whole blood CMV PCR. (a) Qualitative assessment: CMV PCR was performed on 123 serum samples and 74 whole blood samples collected from 132 pregnant women with recent CMV PI. PCR positivity rate was used to calculate sensitivity in serum and whole blood. (b) Quantitative assessment: CMV PCR was performed on 72 paired samples of serum and whole blood collected on the same day from 57 patients. (3) Results: In pregnant women, PCR positivity rate was 89% for serum samples versus 100% in whole blood in the case of very recent PI (<15 days), but only 27% in serum versus 68% in whole blood for PI occurring from 6 weeks to 3 months before. Comparing CMV viral loads between serum and whole blood, we determined the limit of CMV DNA detection in serum as 3 log copies/mL (whole blood equivalent). (4) Conclusions: Serum CMV PCR is reliable in confirming PI in cases when only IgM is detected. It is therefore a valuable tool in introducing valaciclovir treatment as early as possible to prevent mother-to-child CMV transmission. Full article

Very small glaciers (glacierets) react strongly to climatic variations. This is well expressed in their interannual size changes, which are most evident in autumn, at the end of the glacial mass balance year. This study presents results from the detailed research of two very small glaciers in the highest northern part of the Pirin Mountains of Bulgaria: Snezhnika and Banski suhodol. Systematic size measurements of these firn-ice bodies, which started in the 1990s and have been made simultaneously for a period of 13 years, show large inter-annual amplitudes against the background of a decreasing trend in response to climate warming. However, the relations are not straightforward, which is demonstrated when comparing size changes to climate data, including logger data obtained from glacier vicinity. This fact makes predictions for the changes in the local climate of high mountain cirques still relatively uncertain. Full article

Vascular smooth muscle cells (VSMCs) can switch from their contractile state to a synthetic phenotype resulting in high migratory and proliferative capacity and driving atherosclerotic lesion formation. The cysteine-rich LIM-only protein 4 (CRP4) reportedly modulates VSM-like transcriptional signatures, which are perturbed in VSMCs undergoing phenotypic switching. Thus, we hypothesized that CRP4 contributes to adverse VSMC behaviours and thereby to atherogenesis in vivo. The atherogenic properties of CRP4 were investigated in plaque-prone apolipoprotein E (ApoE) and CRP4 double-knockout (dKO) as well as ApoE-deficient CRP4 wildtype mice. dKO mice exhibited lower plaque numbers and lesion areas as well as a reduced content of α-smooth muscle actin positive cells in the lesion area, while lesion-associated cell proliferation was elevated in vessels lacking CRP4. Reduced plaque volumes in dKO correlated with significantly less intra-plaque oxidized low-density lipoprotein (oxLDL), presumably due to upregulation of the antioxidant factor peroxiredoxin-4 (PRDX4). This study identifies CRP4 as a novel pro-atherogenic factor that facilitates plaque oxLDL deposition and identifies the invasion of atherosclerotic lesions by VSMCs as important determinants of plaque vulnerability. Thus, targeting of VSMC CRP4 should be considered in plaque-stabilizing pharmacological strategies. Full article

The evolutionary conserved non-heme Fe-containing protein pirin has been implicated as an important factor in cell proliferation, migration, invasion, and tumour progression of melanoma, breast, lung, cervical, prostate, and oral cancers. Here we found that pirin is overexpressed in human colorectal cancer in comparison with matched normal tissue. The overexpression of pirin correlates with activation of transcription factor nuclear factor erythroid 2 p45-related factor 2 (Nrf2) and increased expression of the classical Nrf2 target NAD(P)H:quinone oxidoreductase 1 (NQO1), but interestingly and unexpectedly, not with expression of the aldo-keto reductase (AKR) family members AKR1B10 and AKR1C1, which are considered to be the most overexpressed genes in response to Nrf2 activation in humans. Using pharmacologic and genetic approaches to either downregulate or upregulate Nrf2, we show that pirin is regulated by Nrf2 in human and mouse cells and in the mouse colon in vivo. The small molecule pirin inhibitor TPhA decreased the viability of human colorectal cancer (DLD1) cells, but this decrease was independent of the levels of pirin. Our study demonstrates the Nrf2-dependent regulation of pirin and encourages the pursuit for specific pirin inhibitors. Full article