We observed the changes in electrical activity, biometric and haemodynamic parameters of hearts in animals with experimental diabetes mellitus (DM). As well the effect of carvedilol, PycnogenolR and its combination with carvedilol on DM heart function was tested. DM was induced by streptozotocin over three sequential days at a dose of 25 mg/kg body weight i.p. We started therapy by suspension of carvedilol, PycnogenolR and their combination for six weeks. Blood pressure was measured using tail cuff plethysmography. ECG, haemodynamic and biometric parameters were measured in isolated hearts perfused according to the Langendorff. DM rats had increased systolic arterial blood pressure, thicker free wall of left ventricle but weakened myocardial contractility compared with controls. In contrast to controls, electrophysiological parameters showed prolonged QT interval and increased incidence of dysrhythmias in DM rats. The PycnogenolR administration induced regression of left ventricular hypertrophy, improved left ventriculi contraction and increased coronary flow; however, it did not improve the electrical activity of the myocardium compared with DM ones. Carvedilol also reversed the myocardial remodelling, shortened the duration of QT interval and suppressed the incidence of dysrhythmias. The common combination of drugs improved biometric and haemodynamic parameters compared with DM animals, however, not so significantly as monotherapy. On the other hand, the combination of carvedilol and PycnogenolR significantly reduced the duration of the QT interval and shortened the incidence of dysrhythmias. We can conclude that the administration of PycnogenolR effectively improved haemodynamic parameters, and carvedilol affected biometric parameters and also electrical parameters in DM animals. We observed the marked synergic effect of the combination of both drugs on the electrical activity of myocardium. This combination shortened the most pathologically prolonged QT interval and reduced the number of dysrhythmias.
Diphyllobothriosis is a fish-borne parasitic zoonosis caused by so-called “broad tapeworms” or “fish tapeworms” of different genera of the order Diphyllobothriidea. Dibothriocephalus l atus (Linnaeus 1758), (syn. Diphyllobothrium latum), is a medically important type species of the genus, whose occurrence in various European regions is either regular, e.g. in the Alpine lakes region, or occasional and sporadic, e. g. in the Danube River region. For the latter, data on the detection of D. latus plerocercoids in the second intermediate fish host (European perch Perca fluviatilis), as well as in definitive hosts (human and dog), in which infection was directly linked to the consumption of infected fish from the Danube, were published more than 50 years ago. In order to assess the current situation, we aimed to find out whether D. latus is present in the natural environment of the Danube River. In total, 700 perch from five sampling sites in the Slovak part of the Danube River were examined. Plerocercoids were not detected in any fish examined, which leads to the conclusion that D. latus is currently not present in the studied aquatic environment.
Ribosomal internal transcribed spacer 2 (ITS2) structure was studied in monozoic tapeworms Caryophyllaeus laticeps (Pallas, 1781) from freshwater bream Abramis brama (L.) and white-eyed bream Ballerus sapa (Pallas) and Caryophyllaeus brachycollis Janiszewska, 1951 from A. brama and Mediterranean barbel Barbus meridionalis (Risso). Homologous intragenomic ITS2 structure (474 bp) was determined for C. brachycollis from both fish hosts and for C. laticeps from A. brama (486 bp). Contrary to this, divergent intragenomic ITS2 copies (ITS2 paralogues) were detected in C. laticeps from B. sapa. They were mostly induced by different numbers of short repetitive motif (TA)n within the sequences, allowing their assortment into two ITS2 variants (457 and 467 bp). Current data represent first information on ITS2 structure/ITS paralogues in the caryophyllidean family Caryophyllaeidae with focus on their applicability in the molecular taxonomy of the genus Caryophyllaeus.
Molecular comparative analysis of eggs of four liver and stomach flukes of cervids and domestic ruminants, Fasciola hepatica, Fascioloides magna, Dicrocoelium dendriticum and Paramphistomum cervi, was performed using a new methodological approach for eggshell disintegration. Eggs of all species were crushed mechanically by the Teflon method (PTFE) without use of chemical reagents and an efficient disruption of eggshell was checked microscopically. The egg suspension was then subjected to DNA isolation and PCR amplification using species-specific primers that annealed to the internal transcribed spacer 2 (ITS2) region of ribosomal DNA. The size of PCR products of individual species corresponded well to the size of amplicons obtained from adult flukes. The results provided evidence that the Teflon method does not destroy the structure of egg DNA, thus making the procedure broadly applicable during coprological examinations. Molecular markers introduced here are particularly important for blanket screening and differentiation of morphologically hardly distinguishable F. hepatica, F. magna and P. cervi eggs.
The combination of multicriteria analysis (MCA), particularly analytic hierarchy process (AHP) and geographic information system (GIS) were applied for transmission risk assessment of Fascioloides magna (Trematoda; Fasciolidae) in south-western Slovakia. Based on the details on F. magna life cycle, the following risk factors (RF) of parasite transmission were determined: intermediate (RFIH) and final hosts (RFFH) (biological factors), annual precipitation (RFAP), land use (RFLU), flooded area (RFFA), and annual mean air temperature (RFAT) (environmental factors). Two types of risk analyses were modelled: (1) potential risk analysis was focused on the determination of the potential risk of parasite transmission into novel territories (data on F. magna occurrence were excluded); (2) actual risk analysis considered also the summary data on F. magna occurrence in the model region (risk factor parasite occurrence RFPO included in the analysis). The results of the potential risk analysis provided novel distribution pattern and revealed new geographical area as the potential risk zone of F. magna occurrence. Although the actual risk analysis revealed all four risk zones of F. magna transmission (acceptable, moderate, undesirable and unacceptable), its outputs were significantly affected by the data on parasite occurrence what reduced the informative value of the actual transmission risk assessment.
The species-specific ribosomal internal transcribed spacer 2 (ITS2) markers were designed for PCR-based molecular differentiation of Fasciola hepatica, Fascioloides magna, Dicrocoelium dendriticum and Paramphistomum cervi, liver and stomach flukes of domestic and free living ruminants. Complete ITS2 sequences were obtained for D. dendriticum and P. cervi, for the later species, ITS2 structure was determined for the first time. Intraspecific variation within geographically distant populations was found to be either very low (F. hepatica; D. dendriticum) or even absent (F. magna; P. cervi). ITS2 regions with the absence of intraspecific polymorphisms but with interspecific sequence heterogeneity were applied for design of speciesspecific primers. The specificity of developed primers was tested on genomic DNA isolated from adult individuals of studied fluke species. Application of the primers is of particular value for molecular differentiation of morphologically hardly distinguishable F. hepatica, F. magna and P. cervi eggs after coprological examinations.
Fascioloidosis of wild and domestic ruminants is caused by giant liver fluke, Fascioloides magna (Trematoda; Fasciolidae). In Slovakia, the parasite is present in the Danube floodplain forests permanent focus for almost 30 years. Here we provide data on 11-year survey of F. magna acquired from 137 red deer (Cervus elaphus) hunted in the southwestern hunting grounds (districts Komárno and Dunajská Streda). Almost 47 % of all examined deer, including males, females and fawns, were infected with F. magna. During the studied period, the prevalence ranged between 33.3 % (2009) and 63.6 % (2007). Prevalence of fascioloidosis varied between sexes and age categories; while the lowest overall prevalence was detected in females (33.3 %), higher values were documented for red deer males (50.6 %) and fawns (43.3 %). A presence of giant liver fluke in studied regions of southwestern Slovakia deserves future attention and ongoing monitoring due to a possible threat of F. magna infection of domestic ruminants in overlapping regions.
Blood pressure (BP) rhythm is exhibited in a circadian pattern regulated by complex system of endogenous factors. Administration of pharmacological treatment at the right time can influence the efficacy of treatment; but while kidneys play significant role in BP regulation, little is known about their role in chronopharmacotherapy. This study aimed to compare differences between morning and evening dosing with valsartan and amlodipine combination in both short-term and long-term settings and to elucidate the role of kidneys in chronopharmacology. Spontaneously hypertensive rats aged between 8 and 10 weeks were daily treated with 10mg/kg of valsartan and 4 mg/kg of amlodipine, either in the morning or in the evening with treatment duration of 1 and 6 weeks. After short-term treatment, only morning treatment group demonstrated significantly better outcomes in terms of BP control when compared to placebo. After long-term treatment, both treatment groups gained superior results in BP control against placebo; however, no significant difference was seen between morning and evening treatment. Interestingly, clock gene expression in kidney has been significantly modulated only in the evening-treated groups, with treatment intensifying the reduced Bmal1 levels, while Per2 expression was less altered. However, no direct relation with the outcomes of the therapy has been observed, suggesting that pharmacotherapy may serve as an independent modulator of peripheral circadian clock in the kidney.
The present study evaluates antihyperglycemic activity of fractionated Pycnogenol® and its ability to improve endothelial dysfunction
in diabetic animals. The aim of this study was to isolate from Pycnogenol® mixture its active compounds and compare
their efficacy on observed parameters. Pycnogenol® mixture was fractioned by re-extracting with petroleum ether, chloroform,
ethyl acetate and butanol, subsequently. Pycnogenol® mixture and fractions (butanolic, water, ethyl acetate) were administered
during 6 weeks to diabetic rats. Blood glucose levels were assessed from the arterio-venous blood at the beginning of experiment
and at the end of experiment. Endothelial dysfunction was evaluated as the contractile responses to phenylephrine and
acetylcholine. The amount of collagen I and III was assessed from thoracic aorta after picrosirius red staining. For the confirmation
of the changes on molecular level, we determinated endothelial NO synthase (eNOS) and heat shock protein 90 (Hsp90)
expression from left ventricle.
Overall, the result suggest, that fractions are not so effective on observed parameters as Pycnogenol® mixture itself, indicating
synergistic effect of the plant constituents.