Genetic diversity and population structure of Culex modestus across Europe: does recent appearance in the United Kingdom reveal a tendency for geographical spread?

In mainland Europe, the mosquito species Culex modestus Ficalbi (1890) is a bridge vector for West Nile virus (WNV) from its natural bird-mosquito cycle to mammals. The present study assessed the genetic diversity of Cx. modestus, as well as related Culex species, using the mitochondrial COI DNA barcoding region and compared this with the population structure across Europe. A haplotype network was mapped to determine genealogical relationships among specimens. The intraspecific genetic diversity within individual Culex species was below 2%, whereas the interspecific genetic divergence varied from 2.99% to 13.74%. In total, 76 haplotypes were identified among 198 sequences. A median-joining network determined from 198 COI sequences identified two major lineages that were separated by at least four mutation steps. A high level of intraspecific genetic diversity was not detected in Cx. modestus in samples submitted from different European populations, which indicates that morphologically identified specimens represent a single species and not a species complex. Therefore, it is deduced that different populations of Cx. modestus will show a similar potential to transmit WNV, lending support to concerns that the population present in southeast England represents a risk of transmission to humans.

Photodynamic Effect of some Phthalocyanines on Enveloped and Naked Viruses.

Activity of three photosensitizing phthalocyanine derivatives was tested for photodynamic inactivation towards two coated and two non-enveloped viruses - bovine viral diarrhea virus (BVDV), influenza virus A(H3N2), poliovirus type 1 (PV-1) and human adenovirus type 5 (HAdV5). In the case of coated viruses, a combination of virucidal and irradiation effects was registered by octa-methylpyridyloxy-substituted Ga phthalocyanine (Ga8) toward BVDV, whereas tetra-methylpyridyloxy-substituted Ga phthalocyanine (Ga4) revealed a marked photoinactivation only. No such effect was observed towards influenza A virus. In contrast, the photoinactivating potential of Ga4 and Ga8 marked very high values on naked viruses, especially on HAdV5, at which both the virucidal as well as the irradiation effects became combined.

[COMBINED APPLICATION OF FETAL FIBRONECTIN AND INSULIN-LIKE GROWTH FACTOR IN PRETERM DELIVERY DIAGNOSIS--OUR RESULTS].

The insulin-like growth factor IGFBP-1 is a binding protein (IBP-1), also known as placental protein (PP12), is encoded in people as IGFBP-1 gene. The IGFBP-1 is an especially vital hormone in the female reproductive physiology. The presence of it in large quantities in the amniotic fluid can be used as a biochemical marker for preterm birth and premature rupture of membranes. Fetal fibronectin is a high-molecular glycoprotein, that is produced during pregnancy and has a role as biological glue, binding the foetus membranes to the endometrium. fFN can be found in the cervicovaginal secretions until 22nd gestational week and later on during the last trimester (1 to 3 weeks before birth). fFN is usually absent between 24th and 34th gestational week. Thirty sixth (30%) of all the 120 females tested had positive fFN test results and were, therefore, at high risk for preterm delivery. Despite the implemented tocolytic therapy, 15 (12.5%) of them delivered before the 37th gestational week. The results from the investigation for the presence of pLGFBP-1 in 120 pregnant women show that 35.8% (43 women) were positive and 15(12,5%) of them delivered before the 37th gestational week despite the implemented tocolytic therapy In both tests, the average gestational age of the premature fetus was 32.5 ÷ 2.8 gestational weeks.

Age dependent changes of arterial wall viscoelasticity.

Viscoelastic characteristics (VEC) of old rat aorta (Wistar, 10 months) were obtained by sinusoidal excitation of intraluminal pressure (p) in cylindrical arterial preparations. The pressure excitation frequency (f(exc)) was swept in the range 3-30 Hz up and down at several mean-pressure levels while response volume oscillations were recorded and resonance curves were plotted. Natural frequency (f(0)), dynamic modulus of elasticity (E') and coefficient of viscosity (beta) were estimated from resonance curves and the dependences of VEC on p were drawn. The results showed that f(0) decreased linearly with p whereas our previous data for young rat aorta (Wistar, 4 months) showed independence of f(0) on p. E' increased nonlinearly with p with the values being higher in comparison to young rat aorta. This means stiffening of rat aorta with age in accordance with the known literature data. beta-values increased linearly with p being higher in comparison to young rat aorta, demonstrative of raised intrinsic friction in the wall. VEC values were higher at decreasing f(exc) suggesting that the direction of excitation sweeping also determines the arterial wall biomechanical behaviour. It could be concluded that blood vessels VEC worsen with age, which endangers the arterial wall integrity, especially at higher intraluminal pressure.

Detection of Theileria orientalis in mosquito blood meals in the United Kingdom.

Theileria spp. are tick-borne protozoan parasites that infect a wide range of wild and domestic animals. In this study, the utility of xenosurveillance of blood-fed specimens of Culiseta annulata for detecting the presence of piroplasms in livestock was investigated. Blood-fed mosquitoes were collected at Elmley National Nature Reserve, Kent, United Kingdom. All specimens were morphologically identified, and DNA barcoding was used to confirm the morphological identification. Both the vertebrate host species and Theileria genome was detected within the bloodmeal by real-time PCR. Sequencing was used to confirm the identity of all amplicons. In total, 105 blood-fed mosquitoes morphologically identified as Cs. annulata were collected. DNA barcoding revealed that 102 specimens were Cs. annulata (99%), while a single specimen was identified as Anopheles messeae. Two specimens could not be identified molecularly due to PCR amplification failure. Blood meal analysis revealed that Cs. annulata fed almost exclusively on cattle at the collection site (n=100). The application of a pan-piroplasm PCR detected 16 positive samples (15.2%) and sequence analysis of the amplicons demonstrated that the piroplasms present in the blood meal belonged to the Theileria orientalis group. This study demonstrates how xenosurveillance can be applied to detecting pathogens in livestock and confirms the presence of Theileria species in livestock from the United Kingdom.

BTEX degradation by fungi.

Aromatic hydrocarbons such as benzene, toluene, ethylbenzene and xylenes (BTEX) are environmental pollutants that cause serious problems in groundwater, in surface water and in soil. Specifically, the coastal area of Bourgas bay (Bulgarian Black Sea coast) is very vulnerable to such pollution as the large petrochemical industry situated there poses a continuing threat to the environment. The potential for degradation of BTEX by fungi was studied. Two fungal strains were used in the experiments carried out in a batch reactor. The fungi were isolated from gasoline-polluted sites and were identified as deuteromycetes belonging to the Cladophialophora (CPH) and Cladosporium (CS) genera. It was found that neither of the fungal strains was able to degrade benzene. Ethylbenzene was easily degraded in all cases; o- and m-xylene were fully degraded by Cladophialophora sp. both as single substrates and in mixtures with toluene, whereas Cladosporium sp. was able to degrade them fully in mixtures with toluene only. p-Xylene was only partially degraded in all tests; however, the experiments with Cladosporium sp. showed better results compared to those with Cladophialophora sp. in this particular case. Ethylbenzene, o- and m-xylene negatively affected toluene removal rate of both fungal strains, which suggests possible competitive substrate interactions. In general, the results obtained with Cladophialophora sp. were better than those with Cladosporium sp., except in the case of p-xylene degradation.

Probabilistic assessment of biodegradability based on metabolic pathways: catabol system.

A novel mechanistic modeling approach has been developed that assesses chemical biodegradability in a quantitative manner. It is an expert system predicting biotransformation pathway working together with a probabilistic model that calculates probabilities of the individual transformations. The expert system contains a library of hierarchically ordered individual transformations and matching substructure engine. The hierarchy in the expert system was set according to the descending order of the individual transformation probabilities. The integrated principal catabolic steps are derived from set of metabolic pathways predicted for each chemical from the training set and encompass more than one real biodegradation step to improve the speed of predictions. In the current work, we modeled O2 yield during OECD 302 C (MITI I) test. MITI-I database of 532 chemicals was used as a training set. To make biodegradability predictions, the model only needs structure of a chemical. The output is given as percentage of theoretical biological oxygen demand (BOD). The model allows for identifying potentially persistent catabolic intermediates and their molar amounts. The data in the training set agreed well with the calculated BODs (r2 = 0.90) in the entire range i.e. a good fit was observed for readily, intermediate and difficult to degrade chemicals. After introducing 60% ThOD as a cut off value the model predicted correctly 98% ready biodegradable structures and 96% not ready biodegradable structures. Crossvalidation by four times leaving 25% of data resulted in Q2 = 0.88 between observed and predicted values. Presented approach and obtained results were used to develop computer software for biodegradability prediction CATABOL.

Antibacterial diterpenic acids from Brazilian propolis.

Four labdane-type diterpenic acids and syringaldehyde were isolated and identified from Brazilian propolis. All the compounds exhibit antibacterial activity. The diterpenes, found for the first time in propolis, are typical for some Araucaria species and thus indicate a possible plant source of Brazilian propolis.

In vitro investigation on the effect of a plant preparation with antiviral activity on the functions of mice phagocyte cells.

A polyphenol extract from the aerial roots of the medicinal plant Geranium sanguineum L. (PC) inhibited the reproduction of influenza viruses type A and B in vitro and in ovo and protected mice from mortality in the experimental influenza infection. The in vivo protective effect was connected with multiple biological activities of the preparation. The present paper focuses on the in vitro effects of the polyphenol extract on the functions of peritoneal and alveolar macrophages and blood polymorphonuclear leucocytes (PMNs), isolated from healthy ICR mice. It was found that PC in doses of 12.5 and 25 microg ml(-1) stimulated the phagocytic activity of peritoneal macrophages and blood PMNs. PC in the same doses did not significantly affect the phagocytic activity of alveolar macrophages, the migration of alveolar and peritoneal macrophages or the adherent activity of PMNs. Used in concentrations of 3.1-25.0 microg ml(-1), PC suppressed spontaneous NO production from peritoneal macrophages, while inducible NO production, provoked by LPS-, Ifn-gamma and LPS + Ifn-gamma inductions was not affected. The cell-toxic concentration of 100 microg ml(-1) increased spontaneous and LPS-inducible NO production. The experimental results demonstrated a stimulating effect of PC on the phagocytic activity of murine PMNs and peritoneal macrophages as well as a beneficial effect of the preparation on spontaneous NO production.

[A radionuclide study of esophageal motility disorders in patients with diabetic polyneuropathy]. / Radionuklidnoe izuchenie motilitetnykh narushenii pishchevoda u bol'nykh s diabeticheskoi polinevropatiei.

The radionuclide method was used to examine transport function of the esophagus in 24 patients suffering from diabetes mellitus and symmetric distal polyneuropathy. 99Tc-sulfocolloid was employed as a radiopharmaceutical agent. Radioactivity movement was visualized by means of a gamma chamber. In 9 patients (37.5%), measurements were made of the prolonged transit time for the whole esophagus as well as the prolonged transit time for the interior, median and upper third of the esophagus. The same patients demonstrated a delay of the rate of evacuation of the entire esophagus and respective parts thereof. The radionuclide method described is rapid and noninvasive. It may be included into the diagnostic complex for early identification of vegetative disorders in patients suffering from diabetic polyneuropathy.

[Radionuclide study of disorders of esophageal motility in patients with diabetic polyneuropathy]. / Radionuklidnoe izuchenie dvigatel'nykh narushenii pishchevoda u bol'nykh s diabeticheskoi polinevropatiei.

Radionuclide method was used to examine transport function of the esophagus in 24 patients with diabetes mellitus and symmetric distal polyneuropathy. 99mTc-+sulfur colloid was employed as a radiopharmaceutical agent. The movement of radioactivity was elucidated visually with the aid of a gamma-chamber. In 9 patients, measurements were made of the prolonged transit time of the whole esophagus as well as prolonged transit time for the lower, median and upper third of the esophagus. The same patients manifested deceleration of the evacuation of the entire esophagus and respective parts thereof. That method can be included into the diagnostic complex intended for early diagnosis of vegetative disorders in patients with diabetic polyneuropathy and for evaluating the intensity of autonomous neuropathy.