Circulatory diseases and the wide sex and ethnic life expectancy gaps in Bulgaria since 2010.

OBJECTIVES: Despite extensive public health initiatives, Bulgaria still has the lowest life expectancy at birth (LE) in the European Union. Sex and ethnic differences in LE and mortality are also exceptionally large. We aimed to identify what causes of death drive these wide disparities and thus provide clear targets for future public health interventions. DESIGN AND METHODS: We conducted a retrospective analysis of mortality rates from 2010 to 2022 to assess sex disparities in LE by age and cause of death. Combining mortality data with the 2021 Bulgarian census also allowed us to study LE disparities among the three main ethnic groups (Bulgarians, Turks, and Roma). We implemented standard demographic decomposition methods to quantify the role of seven major causes of death on LE disparities. RESULTS: We found that the difference between male and female LE has persisted for around seven years. Circulatory diseases contribute 3.66 years, or around 50% of the male-female gap. Ethnic LE disparities are larger for women than for men. Circulatory diseases account for more than 60% of these ethnic LE gaps. COVID-19 mortality explained between 0.5 and 1.1 years of the male-female gap. We found minimal differences in COVID-19 mortality across ethnic groups in Bulgaria. CONCLUSION: In Bulgaria, circulatory diseases contributed more to both the sex and ethnic LE gaps than in any other previously studied country. Our findings suggest that future public health policy initiatives should focus on circulatory diseases to narrow the Bulgarian LE disparities. One possible target for such a policy would be to reduce excessive smoking and alcohol consumption.

A rapid differentiation between oak honeydew honey and nectar and other honeydew honeys by NMR spectroscopy.

An NMR-based approach for rapid differentiation of oak honeydew honey from all other honey types (floral and other honeydew honeys) was proposed. It is based on the identification of the signals of the protons and the carbon of the methylene group of quercitol in the (1)H and (13)C NMR spectra of honey. The presence of quercitol was supported by TOCSY spectroscopy. Quercitol is a deoxyinositol which is regarded as a good taxonomic marker for the genus Quercus. All samples of oak honeydew honey contained quercitol, while in floral honey samples and honeydew honeys from fir and spruce it was absent. In addition, the described approach is promising with respect to quercitol quantification in honey by qNMR.

Seasonal variations in the chemical composition of vine-grape leaf surface.

Surface leaf metabolites of two seedlings of a Bulgarian winemaking cultivar Storgozia I(1) were analyzed in two seasons - summer (July) and autumn (October). The resistance towards some fungal pathogens of one of the plants was estimated as superior to the resistance of the other one. Significant seasonal variations in the chemical constituents of the two seedlings were observed. The main metabolites of the summer samples were sterols, terpenes, fatty acids and heterocyclic compounds. In autumn, sterol and fatty acid contents decreased, mono- and diterpenes and heterocyclic compounds disappeared and instead of them hydrocarbons and alcohols were detected. Some individual components - stearic acid, alpha-amyrin, lupeol and squalene - correlated with the estimated resistance and were therefore proposed as biomarkers for the fungal resistance in grape-vine leaves.

Raman scattering of Si matrix with randomly distributed nanoparticles of semiconducting silicides.

Samples, representing Si matrix with nanoformations of the semiconducting silicides beta-FeSi2 and Mg2Si are studied by Raman scattering. The unpolarized Raman spectra of the samples are interpreted in the framework of the appearance of interface-phonon polaritons. The theoretical dispersion relations of the interface-phonon polaritons in the system Si/silicide/Si are obtained from the Maxwell equations. The correspondence of the theoretical calculations and the experimental observations appeared to be sufficiently good. An evolution of the features in the Raman spectra on the experimental conditions is observed.

Improved phytoaccumulation of cadmium by genetically modified tobacco plants (Nicotiana tabacum L.). Physiological and biochemical response of the transformants to cadmium toxicity.

The response of tobacco plants (Nicotiana tabacum L.)--non-transformed and transformed with a metallothionein gene MThis from Silene vulgaris L.--to increase cadmium supply in the nutrient solution was compared. The transgenic plants accumulated significantly more Cd both in the roots and the leaves. Visual toxicity symptoms and disturbance in water balance were correlated with Cd tissue content. Treatment with 300 microM CdCl(2) resulted in inhibition of photosynthesis and mobilization of the ascorbate-glutathione cycle. Treatment with 500 microM CdCl(2) led to irreversible damage of photosynthesis and oxidative stress. An appearance of a new peroxidase isoform and changes in the leaf polypeptide pattern were observed at the highest Cd concentration. The level of non-protein thiols gradually increased following the Cd treatment both in transgenic and non-transformed plants.

Multiple origins of cultivated grapevine (Vitis vinifera L. ssp. sativa) based on chloroplast DNA polymorphisms.

The domestication of the Eurasian grape (Vitis vinifera ssp. sativa) from its wild ancestor (Vitis vinifera ssp. sylvestris) has long been claimed to have occurred in Transcaucasia where its greatest genetic diversity is found and where very early archaeological evidence, including grape pips and artefacts of a 'wine culture', have been excavated. Whether from Transcaucasia or the nearby Taurus or Zagros Mountains, it is hypothesized that this wine culture spread southwards and eventually westwards around the Mediterranean basin, together with the transplantation of cultivated grape cuttings. However, the existence of morphological differentiation between cultivars from eastern and western ends of the modern distribution of the Eurasian grape suggests the existence of different genetic contribution from local sylvestris populations or multilocal selection and domestication of sylvestris genotypes. To tackle this issue, we analysed chlorotype variation and distribution in 1201 samples of sylvestris and sativa genotypes from the whole area of the species' distribution and studied their genetic relationships. The results suggest the existence of at least two important origins for the cultivated germplasm, one in the Near East and another in the western Mediterranean region, the latter of which gave rise to many of the current Western European cultivars. Indeed, over 70% of the Iberian Peninsula cultivars display chlorotypes that are only compatible with their having derived from western sylvestris populations.

Chlorsulfuron resistant transgenic tobacco as a tool for broomrape control.

Broomrape (Orobanche ramosa L.) is the most important parasitic plant that infests tobacco (Nicotiana tabacum L.). Chemical treatment of the soil is not effective and crop rotation is not acceptable to solve this problem because of the long viability period of Orobanche seeds in the soil. Application of systemic herbicides in the field with herbicide resistant tobacco could be a successful tool for broomrape control. Several tobacco cultivars were transformed with a mutant ahas3R gene for resistance to the herbicide chlorsulfuron (Glean, DuPont). Transformed plants were selfed and the segregation of resistance was followed in the next generation. The efficiency of the herbicide was demonstrated in greenhouse and field trials. An Orobanche/tobacco growth system was used in order to prove the lethal effect of the herbicide to the attached broomrape plants.

Microsatellite analysis of Rosa damascena Mill. accessions reveals genetic similarity between genotypes used for rose oil production and old Damask rose varieties.

Damask roses are grown in several European and Asiatic countries for rose oil production. Twenty-six oil-bearing Rosa damascena Mill. accessions and 13 garden Damask roses were assayed by molecular markers. Microsatellite genotyping demonstrated that R. damascena Mill. accessions from Bulgaria, Iran, and India and old European Damask rose varieties possess identical microsatellite profiles, suggesting a common origin. At the same time, the data indicated that modern industrial oil rose cultivation is based on a very narrow genepool and that oil rose collections contain many genetically identical accessions. The study of long-term vegetative propagation of the Damask roses also reveals high somatic stability for the microsatellite loci analyzed.

Assessment of polysomaty, embryo formation and regeneration in liquid media for various species of diploid annual Medicago.

To avoid polyploidy in regenerants the source of explant material should be monosomatic. Therefore, the leaf and petiole tissue of five diploid Medicago species (Medicago ciliaris, Medicago murex, Medicago orbicularis, Medicago polymorpha and Medicago truncatula cv. Jemalong, and the ecotype R108-1) was assessed for polysomaty by flow cytometry. For the species studied the frequency of 2C nuclei was about 90% in leaves compared with that in petioles. Embryos were readily formed from tissue of leaves in liquid media containing 1 mg l(-1) or 4 mg l(-1) dichlorophenoxyacetic acid (2,4-D). For embryo development two procedures were tested - prolonged use of induction medium and treatment with polyethylene glycol Mw 6000 (PEG). The highly regenerable genotypes M. truncatula cv. Jemalong and R108-1 showed efficient conversion of embryos after maturation in liquid medium. The regenerated plants were diploid and with normal phenotype.

Variability of the N-protein and the intergenic region of the S RNA of tomato spotted wilt tospovirus (TSWV).

The genetic heterogeneity of the N protein gene and the intergenic region (IGR) of the S RNA from tomato spotted wilt virus (TSWV) isolates, collected in Bulgaria, were compared with isolates from other parts of the world. The results substantiated the highly conserved nature of the N protein. Twenty six independent sequences revealed only seven variable amino acid positions, common to all isolates. The type of amino acids present in these positions seems to be independent of the geographical origin. In contrast to the structural N protein, comparisons of the related IGR-sequences led to clusters correlated with the geographical origin of the isolates. Although the overall sequence homology in the IGRs was much lower than for the N proteins, three conserved parts within this region were identified. The outstanding part was a central area of 31 nucleotides with a significantly increased GC-content. This was located in both viral- and viral-complement RNA at structures with similar foldings, which led to the assumption that this stabilised structure, rather than a sequence motive, might serve as a transcription terminator during the synthesis of the two mRNAs from the ambisense segments.

New members of a cold-responsive group-3 Lea/Rab-related Cor gene family from common wheat (Triticum aestivum L.).

A Cor (cold-responsive) cDNA that belongs to the group-3 Lea (late embryogenesis abundant)/Rab (responsive to abscisic acid, ABA) family was isolated from a winter-hardy cultivar of common wheat (Triticum aestivum L.). Screening of a cold-acclimated cDNA library was performed using an ABA- and other stress-responsive barley cDNA clone, Hva1, as a probe. A wheat cDNA clone (designated as Wrab19) putatively encoded a basic (pI = 10.3) and hydrophobic protein with 179 amino acids. The deduced protein showed characteristics of the group-3 LEA/RAB protein family. In contrast to the single copy barley Hva1, Wrab19 belonged to a multigene family in the hexaploid wheat genome and six loci were assigned to the homoeologous group 1 chromosomes. Using Wrab19 as a probe, four homologous cDNAs (designated as Wrab17) were isolated that encoded acidic (pI = 4.6-4.7) and hydrophobic proteins, all with 166 amino acids. The deduced proteins showed high homology (a mean of 84% identity) with a barley gibberellic acid (GA3)-inducible protein, ES2A, and several other group-3 LEA/RAB proteins. Wrab17 was considered to be a three-copy gene and each copy was assigned to chromosome 5A, 4B or 4D of hexaploid wheat. Transcripts of both Wrab19 and Wrab17 accumulated within 1 day of cold acclimation at 4 degrees C. They were responsive to ABA and/or GA3, but showed some cultivar differences in their response to these plant hormones. We conclude that the two genes are new members of the group-3 Lea/Rab-related Cor gene family in wheat.

A cold-responsive wheat (Triticum aestivum L.) gene wcor14 identified in a winter-hardy cultivar 'Mironovska 808'.

A cDNA library was constructed from a cold-acclimated winter-hardy common wheat (Triticum aestivum L.) cultivar 'Mironovska 808'. Using this library and a cold- and light-responsive barley cDNA clone cor14b as a probe, cDNAs of a homologous wheat gene wcor14 were isolated. Two identical cDNAs designated as wcor14a had an open reading frame encoding an acidic (pI = 4.71) and hydrophobic polypeptide with 140 amino acids (MW = 13.5 kDa). The deduced WCOR14a polypeptide showed 70% identity with the barley chloroplast-imported COR14b and had a nearly identical N-terminal, putative chloroplast transit peptide of 51 amino acid residues. Another cDNA clone wcor14b was assumed to encode a polypeptide WCORb which had 5 substitutions and a frame shift in the C-terminal region as compared with WCOR14a. RACE PCR, genomic PCR and Southern blot analyses suggested that wcor14 and its related sequences constitute a small multigene family with and without an intron in the hexaploid wheat genome. Northern blot analysis showed that transcripts of wcor14 accumulated within 3-6 hours of cold acclimation at 4 degrees C and the level reached a maximum at day 3. The transcripts became non-detectable within 3 hours after de-acclimation at room temperature. Contrary to the barley cor14b, a similar level of wcor14 transcripts was detected under the continuous darkness. Neither treatment with NaCl, ABA nor dehydration induced its expression. Based on these results we conclude that wcor14 is a wheat orthologue of the barley cor14b and specifically induced by low temperature.

Relevance between arthroscopic pathology and clinical characteristics in knee osteoarthritis.

UNLABELLED: Arthroscopy can be used to identify accurately cartilage and synovial changes in knee osteoarthritis (OA). The objective of this study was to find the correlation between arthroscopically established knee chondropathy and synovitis and the corresponding algofunctional characteristics in OA. METHODS: 41 patients with OA in only one knee joint were investigated. The diagnosis of knee OA fulfilled the clinical, laboratory and radiological ARA criteria. Low pressure arthroscopy of the affected knee was performed under local anaesthesia. Chondropathy was evaluated according to the method of R E Outerbridge. Synovial inflammation was assessed by the scoring system of S. Lindblad and E. Hedfors. RESULTS: We found statistically significant correlation (Spearman's rho test) between chondropathy of the knee and the following clinical indices: pain at night, pain after standing 30 minutes, pain on walking, pain on getting up from a chair without help of arms, maximum walking distance. We failed to demonstrate statistical significant correlation between chondropathy and morning stiffness and activities of daily living. There was also no statistical correlation between knee synovitis and any of the Lequesne's clinical indices. CONCLUSION: The arthroscopically established scores of the cartilage pathology were consistent with most of the clinical algofunctional symptoms. The degree of synovitis in our study was not correlated with clinical activity symptoms of OA, according to Leguesne. Five positive algofunctional Lequesne's scores corresponded to heavy breakdown of knee cartilage in OA.

Expression of an anther-specific chalcone synthase-like gene is correlated with uninucleate microspore development in Nicotiana sylvestris.

Two cDNA clones, specifically expressed in Nicotiana sylvestris anthers during uninucleate microspore development, were isolated using a subtractive hybridization approach. Sequence analysis showed that one of them, NSCHSLK, displayed a high level of similarity to several anther-specific chalcone synthase-like (CHSLK) proteins and an ORF from chromosome I of Arabidopsis thaliana. A lower, but significant, similarity to chalcone synthases and closely related enzymes (CHSRE) was also detected. The structure of the nschslk gene was found to be typical of the chalcone (chs)/stilbene (sts) synthase family. Expression of NSCHSLK mRNA was confined to microspores and tapetal cells. UV-irradiation or infection with Phytophthora parasitica var. nicotianae of transgenic Nicotiana benthamiana plants carrying a chimeric nschslk/GUS gene indicated that the nschslk promoter exhibits the same anther-specific, developmentally regulated expression pattern. Comparison of CHSRE and CHSLK polypeptide sequences revealed some important similarities and differences between the two groups. The data presented in this study, suggest that the anther-specific chslk genes represent a separate sub-family of plant polyketide synthases related to chs/sts in terms of gene structure, polypeptide sequence and the possible catalytic mechanism, but differing in substrate/product specificity. The putative role of CHSLK enzymes in anther development and particularly in exine synthesis is discussed.

Arthrosonography in the evaluation of osteoarticular and soft-tissue structures in rheumatology.

BACKGROUND: Real-time ultrasonography is a widely available, reliable, diagnostically sensitive and specific noninvasive modality which allows measuring of the thickness of different anatomical landmarks which form joints, of the periarticular soft tissue and connective tissue structures, of their echogenicity and the amount of joint effusion. The present study was designed to assess the clinical and diagnostic applicability of high-frequency ultrasonography in rheumatology. METHODS: We used SONOACE 1500 (Medison Europe GmbH) with a 7.5 MHz transducer. RESULTS: We demonstrate the most characteristic ultrasonographic patterns which have diagnostic significance in the seven most common rheumatologic disorders. DISCUSSION: The pathological ultrasonographic patterns of the joints, periarticular and soft-tissue structures in rheumatologic disorders we present give an insight into the clinical applicability of arthrosonography and its inherent advantages over the conventional radiological examination--namely, ultrasonography is noninvasive, uses no radiation, which allows reproducibility and follow-up, widely available and relatively inexpensive and as such can be performed at bedside or on an outpatient basis.

Cloning novel alfalfa cyclin sequences--a RACE-PCR approach.

Cyclins are a complex group of proteins involved in regulation of the eukaryotic cell division cycle via their interaction with cyclin dependent kinases (Cdks). Cyclin gene sequences have been cloned from a number of plant species, including alfalfa, but the diversity of these genes suggests that there are many plant cyclins which have yet to be characterized. A RACE-PCR strategy has been adopted for cloning cyclin gene sequences expressed during direct somatic embryogenesis in alfalfa. RT-PCR with nested degenerate primers was used to amplify the highly conserved "cyclin box" region of a novel A-like cyclin mRNA sequence expressed after induction of somatic embryogenesis. The sequence of this PCR product was used to design primers for 5'- and 3'-RACE protocols. 5'-RACE using a modified SLIC (single strand ligation to single stranded cDNA) procedure revealed considerable sequence heterogeneity in the N-terminal region of the coding sequence with several closely related sequences apparent. Conventional 3'-RACE generated a single cyclin sequence. The complete coding sequence of one member of this A-like cyclin subgroup has been obtained by this RACE strategy and confirmed by PCR amplification and sequencing of alfalfa genomic DNA.

Blood gases, electrolytes and metabolic monitoring in children with acute failure of vital functions.

The priority of direct monitoring of blood gases in Paediatric Intensive Care Units (PICU) increased substantially after introduction of the Deep Picture method and Oxygen Status Algorithm (OSA) (1) into medical practice. We used the advantages of these methods as a prerequisite for a more detailed and deeper analysis of the blood oxygen profile (2, 3). The aims of the present paper were: 1. To illustrate the applicability of the capacity coefficients beta 1.0, beta 2.3, beta 5-4 of the transported oxygen and the "Useful Ratio" (UR) index of the haemoglobin oxygen, previously described by us, and the benefit derived from differentiation of the states of hyperoxia, normoxia and hypoxia; hyperoxaemia, normoxaemia and hypoxaemia on the Blood Oxygen Binding Curve (BOBC) in critically ill newborns, infants and children. 2. To expand the diagnostic capacity of the Blood Gas Map (BGM) used with the OSA in children and to supplement the arterial oxygen diagnostics with new indices that reflect the relationship between oxygen uptake and oxygen transported in the body. 3. To share our experience in PICU related to the acid-base-electrolytes relationship and to the possibility of assessing the reno-hepatic regulation according to the changes of the acid-base status in critically ill children.

Effect of 2,4-D precultivation on regeneration capacity of cultivated barley.

A reliable protocol for regeneration of barley from seedling explants including leaf bases with the complete apical meristem has been developed. Callus induction and shoot regeneration was only achieved when 28 days-old-seedlings were grown in the presence of high concentrations of 2,4-D. The Bulgarian barley variety Ruen commercially important for brewery industry was used as a model. The regeneration response of a range of Bulgarian barley genotypes was evaluated. Cytological analyses were performed at the level of callus and regenerants.

In vitro production of haploid plants.

Although several methods have been developed for producing haploid plants, the in vitro techniques are much more efficient than inter-specific hybridization or treatment with plant-growth regulators, temperature or irradiation. Androgenesis is the most universal of these techniques but ovule culture and the bulbosum method could complement or replace anther culture in those species or genotypes with less responsive male gametes. Genotype, environment, physiological status of the donor plant, and culture conditions and components all need to be taken into account when developing procedures for producing haploid and dihaploid plants. Suitable methods are already well established for a number of important crops. However, many problems, related to regeneration frequency, gametoclonal variation and albinism, are still unsolved. It is now clear that haploids and dihaploids form the ideal system for genetic manipulation in plants. Their key role in producing new theoretical and applied knowledge in plant science is an important aspect of our review.