Simulium reptans (Linnaeus, 1758) and Simulium reptantoides Carlsson, 1962 from the Balkan Peninsula

Simulium reptans (Linnaeus, 1758) and Simulium reptantoides Carlsson, 1962 are two species of the Simulium reptans group whose distribution is unclear because of their confusing taxonomy and systematics. Their genetic variability is well known for populations in northern and central Europe and shows that both species have two forms; however, the genetic variability of these species in southern and eastern Europe is unknown. To identify the status of these two species in southeast Europe, mtDNA was extracted from 19 individuals from 12 localities across the Balkan Peninsula. Phylogenetic analysis confirmed the existence of two species with 7.38–7.94% divergence. Each species was comprised of two clades, with 2.31% and 1.43% interclade divergence for S. reptans and S. reptantoides , respectively. This study revealed the presence of both species across the Balkans and that S. reptans occurs in this area in only one form ( S. reptans B), while S. reptantoides is found in two genetic forms (A and B)

Bioindication potential of physiological responses of gypsy moth Lymantria dispar L. (Lepidoptera) on the benzo[a]pyren effects

Intenzivan antropogeni uticaj u poslednjih nekoliko decenija doveo je do značajnog porasta stepena zagađenja biosfere, kojem u velikoj meri doprinosi toksični i kancerogeni organski polutant - policiklični aromatični ugljovodonik, benzo[a]piren. Cilj ove doktorske disertacije bio je procena odgovora fizioloških i molekularnih parametara larvi Lymantria dispar kao potencijalnih biomarkera sredinskog zagađenja benzo[a]pirenom. Ispitan je hronični uticaj dve koncentracije ovog ksenobiotika (5 i 50 ng/g suve mase hrane) na larve L. dispar iz legala sakupljenih u nezagađenoj šumi (planina Majdanpek) i sakupljenih u zagađenoj šumi (okolina rudnika Bor D.O.O), sa svrhom procene efekta porekla populacije. Larve iz nezagađene šume pokazuju pad specifične aktivnosti katalaze u srednjem crevu, ali i značajan porast aktivnosti antioksidativnih enzima - superoksid dismutaze i katalaze, na nivou hemolimfe, dok je u populaciji iz zagađene šume zabeleženo povećanje aktivnosti ova dva enzima samo u okviru srednjeg creva. Detoksifikacioni enzimi, karboksilesteraze, pokazuju osetljivost i povećanje aktivnosti u prisustvu benzo[a]pirena, u srednjem crevu larvi iz nezagađene šume, kao i u mozgu larvi iz zagađene šume, koje pored toga, na nivou srednjeg creva odlikuje značajna indukcija aktivnosti glutation S-transferaza. Acetilholinesteraze nisu osetljive na primenjene koncentracije ovog ksenobiotika. Povećanje ekspresije proteina Hsp70 u okviru srednjeg creva uočeno je kod obe populacije larvi, a na nivou mozga jedino u populaciji iz zagađene šume i to pri dejstvu niže koncentracije benzo[a]pirena. Većina digestivnih enzima nije osetljiva na prisustvo benzo[a]pirena sa izuzetkom tripsina, alkalnih fosfataza i lipaza, koje odlikuje povećanje aktivnosti kod larvi iz nezagađene šume, a larve iz zagađene šume karakteriše jedino indukcija lipazne aktivnosti. Za populaciju larvi iz nezagađene šume svojstven je pad koncentracije ukupnih lipida u hemolimfi i nepromenjen sadržaj trehaloze, dok ovi parametri ne pokazuju promene u populaciji iz zagađene šume. Hronično prisustvo benzo[a]pirena i produženo angažovanje energetski zahtevnih, odbrambenih mehanizama, odrazilo se na komponente fitnesa larvi L. dispar pri čemu se populacija iz nezagađene šume pokazala manje podložnom energetskom deficitu, ispoljavajući smanjenje mase i relativne brzine rasta larvi petog larvenog stupnja, dok larve iz zagađene šume, pored iste promene prethodna dva parametra fitnesa, odlikuje i produženo razviće. Uočeno je da su odgovori antioksidativnih enzima - superoksid dismutaze i katalaze, u hemolimfi, kao i karboksilesteraza i digestivnih enzima - tripsina i alkalnih fosfataza, u srednjem crevu larvi, specifični i potencijalno primenjivi biomarkeri zagađenosti životne sredine sa benzo[a]pirenom za populaciju larvi poreklom iz nezagađene šume. Sličnu primenu, kada je u pitanju populacija larvi iz zagađene šume, mogu naći specifične aktivnosti superoksid dismutaze, katalaza i glutation S-transferaza u srednjem crevu larvi, aktivnost karboksilesteraza i ekspresija Hsp70 u mozgu larvi L. dispar.Over the last few decades intense anthropogenic influence has led to a significant increase of the pollution levels in the biosphere, which can greatly be attributed to the toxic and carcinogenic organic pollutants - polycyclic aromatic hydrocarbones, and especially to the benzo[a]pyrene. The aim of this dissertation was to evaluate responses of the physiological and molecular parameters of the Lymantria dispar larvae as a potential biomarkers of environmental pollution by benzo[a]pyrene. For the purpose of assessing the effects of population origin, L. dispar egg-masses were collected from unpolluted forest (Majdanpek mountain) and polluted forest (vicinity of Bor D.O.O mine). The chronic effects of the two benzo[a]pyrene concentrations in the diet (5 and 50 ng/g dry food weight) on these two population of larvae were investigated. Larvae from unpolluted forest showed a decrease of the catalase specific activity in the midgut, and a significant increase in the activity of antioxidative enzymes - superoxide dismutase and catalase, in the hemolymph, while the population from the polluted forest expressed an increase in the activity of these two enzymes only within the midgut. Detoxification enzymes - carboxylesterases, showed sensitivity and increase in activity in the presence of benzo[a]pyrene in the midgut of larvae from unpolluted forest, as well as in the brain of larvae from polluted forest, which in addition, are characterized by significant induction of glutathione S-transferase activity at the midgut level. Acetylcholinesterases are non inducible to the applied concentrations of this xenobiotic. An increase in expression of Hsp70 protein at midgut level was observed in both larval populations, while only the population from contaminated forest showed rise in brain Hsp70 concentration, after treatment with lower dose of benzo[a]pyrene. Most digestive enzymes are insensitive to the presence of benzo[a]pyrene with the exception of trypsin, alkaline phosphatases, and lipases which demonstrated increased activity in larvae from unpolluted forest. Larvae from polluted forest exibited only induction of lipase activity. The population of non-polluted larvae is characterized by a decrease in the concentration of total lipids in the hemolymph and an unchanged trehalose content, while these parameters do not show changes in the population from the contaminated forest. The chronic presence of benzo[a]pyrene and prolonged engagement of energy-demanding defense mechanisms have reflected on the fitness components of L. dispar larvae, with population from unpolluted forest showing less susceptibility to energy deficiency and exhibiting reduced mass and relative growth rates of the fifth instar larvae, while the larvae from the polluted forest, in addition to the same changes in previous two fitness parameters, have also showed prolonged development. It have been noticed that responses of the antioxidative enzymes - superoxide dismutase and catalase in hemolymph, as well as carboxylesterases and digestive enzymes - trypsin and alkaline phosphatases, from the larvae midgut, are specific and potentially applicable biomarkers of environmental contamination with benzo[a]pyrene for larval populations originally from unpolluted forest systems. Similarly, when it comes to larval populations from contaminated forests, specific activities of superoxide dismutase, catalase and glutathione S-transferases in the larvae midgut, as well as carboxylesterase activity and expression of Hsp70 in the brain of L. dispar larvae, could be potentially used in biomonitoring

Douglas fir impact of the dynamics and composition of humus in the soil of indigenous beech forest in Western Serbia

This study investigates the impact of organic matter from Douglas fir (Pseudotsuga menziesii [Mirb.] Franco) on the amount and composition of humus in acid brown soil in a climatoregional beech forest (Fagetum moesiacae montanum B. Jov. 1967 s.l.) on Mt. Maljen. To accomplish this objective, we performed a one-year litterbag decomposition experiment with litterfall from Fagus moesiaca and Pseudotsuga menziesii. The quantitative and qualitative content of humus and the intensity of the decomposition process of organic mat-ter from beech and Douglas fir were analysed. Less humus was found during the experiment under Douglas fir than under autochthonous beech at the control site, as well as a decreasing trend for humus levels and quality (unfavourable chemical composition). It was concluded that these changes, caused by the effects of the clearcutting of beech and, in the future, of Douglas fir, and the slower decomposition of organic matter from Douglas fir will contribute to further degradation of the beech habitat on Mt. Maljen in terms of productivityУ раду је истраживан утицај органске материје дуглазије (Pseudotsuga menziesii [Mirb.] Franco) на количину и састав хумуса киселог смеђег земљишта климарегионалне букове шуме (Fagetum moesiacae montanum B. Jov. 1967 s.l.) на Маљену. За постизање наведеног циља обављен је једногодишњи in situ експеримент, коришћењем “litter bag” методе/технике са стељом букве (Fagus moesiaca) и дуглазије (Pseudotsuga menziesii). Анализиран је квантитативан и квалитативан садржај хумуса и интезитет процеса разлагања органске материје букве и дуглазије. Утврђена је нижа количина хумуса, уочен је тренд смањивања количине хумуса током истраживања и опадање квалитета хумуса (неповољнији хемијски састав) у култури дуглазије у односу на контролну површину под аутох-то ном буквом. Закључено је да ће ове промене, изазване ефектима чисте сече бу кве и у будућности дуглазије, и успорено разлагање органске материје дуглазије, до принети даљој деградацији буковог станишта у погледу продуктивности на пла нини Маљен

Antibacterial activity of griseofulvin analogues as an example of drug repurposing

Griseofulvin is a well-known antifungal drug that was launched in 1962 by Merck & Co. for the treatment of dermatophyte infections. However, according to predictions using the Way2Drug computational drug repurposing platform, it may also have antibacterial activity. As no confirmation of this prediction was found in the published literature, this study estimated in-silico antibacterial activity for 42 griseofulvin derivatives. Antibacterial activity was predicted for 33 of the 42 compounds, which led to the conclusion that this activity might be considered as typical for this chemical series. Therefore, experimental testing of antibacterial activity was performed on a panel of Gram-positive and Gram-negative micro-organisms. Antibacterial activity was evaluated using the microdilution method detecting the minimal inhibitory concentration (MIC) and the minimal bactericidal concentration (MBC). The tested compounds exhibited potent antibacterial activity against all the studied bacteria, with MIC and MBC values ranging from 0.0037 to 0.04 mg/mL and from 0.01 to 0.16 mg/mL, respectively. Activity was 2.5–12 times greater than that of ampicillin and 2–8 times greater than that of streptomycin, which were used as the reference drugs. Similarity analysis for all 42 compounds with the (approximately) 470,000 drug-like compounds indexed in the Clarivate Analytics Integrity database confirmed the significant novelty of the antibacterial activity for the compounds from this chemical class. Therefore, this study demonstrated that by using computer-aided prediction of biological activity spectra for a particular chemical series, it is possible to identify typical biological activities which may be used for discovery of new applications (e.g. drug repurposing)

Seed oil and seed oil byproducts of common purslane (Portulaca oleracea L.): A new insight to plant-based sources rich in omega-3 fatty acids

In the present study, nutritional value, chemical composition and bioactive properties of purslane seeds, seed oils and seedcakes were examined. Data were analyzed by a one-way ANOVA, while means were compared with Tukey's HSD test. For seed oil extraction mechanical and ultrasound assisted methods were tested. Cold extraction methods (CE1 and CE2) resulted in higher oil yield (increased by 33.7% and 38.1%, respectively) comparing to hot extraction (HE) method. Seeds contained the highest amount of fats and energy (15.03 ± 0.06 g/100 g dry basis (db) and 459 ± 1 kcal/100 g db, respectively), while seedcakes from CE2 had the highest content in proteins and ash (31.20 ± 0.03 and 4.27 ± 0.06 g/100 g db, respectively). Seeds and seedcakes contained a balanced content of linoleic and α-linolenic acids (33.80–34.74% and 32.83–34.64%, respectively). HE and CE1 oils had slightly higher amounts of α-linolenic (39.67% and 39.57%, respectively) than linoleic acid (35.44% and 35.13%, respectively), whereas CE2 oils contained twice as much linoleic as α-linolenic acid (49.77% and 24.18%, respectively). In conclusion, the tested materials are good sources of omega-6 and omega-3 fatty acids and proteins, while extraction method affected oil yield and fatty acids composition of seed oils

Sex differences in Tfh cell help to B cells contribute to sexual dimorphism in severity of rat collagen-induced arthritis.

The study examined germinal centre (GC) reaction in lymph nodes draining inflamed joints and adjacent tissues (dLNs) in male and female Dark Agouti rat collagen type II (CII)-induced arthritis (CIA) model of rheumatoid arthritis. Female rats exhibiting the greater susceptibility to CIA mounted stronger serum CII-specific IgG response than their male counterparts. This correlated with the higher frequency of GC B cells in female compared with male dLNs. Consistently, the frequency of activated/proliferating Ki-67+ cells among dLN B cells was higher in females than in males. This correlated with the shift in dLN T follicular regulatory (Tfr)/T follicular helper (Tfh) cell ratio towards Tfh cells in females, and greater densities of CD40L and CD40 on their dLN T and B cells, respectively. The higher Tfh cell frequency in females was consistent with the greater dLN expression of mRNA for IL-21/27, the key cytokines involved in Tfh cell generation and their help to B cells. Additionally, in CII-stimulated female rat dLN cell cultures IFN-γ/IL-4 production ratio was shifted towards IFN-γ. Consistently, the serum IgG2a(b)/IgG1 CII-specific antibody ratio was shifted towards an IgG2a(b) response in females. Thus, targeting T-/B-cell interactions should be considered in putative further sex-based translational pharmacology research

MIF and insulin: Lifetime companions from common genesis to common pathogenesis

Pro-inflammatory nature of macrophage migration inhibitory factor (MIF) has been generally related to the propagation of inflammatory and autoimmune diseases. But this molecule possesses many other peculiar functions, unrelated to the immune system, among which is its supportive role in the post-translational modifications of insulin. In this way MIF enables proper insulin conformation within the pancreatic beta cell and its full activity. The inherent or acquired changes in MIF expression might therefore lead to different insulin processing and initiation of autoimmunity. The relation between MIF and insulin does not stop at this point; these two molecules continue to interact during pathological states characterized by inflammation and insulin resistance. In this context, MIF indirectly and negatively influences insulin action by boosting inflammatory environment and disabling target cells to respond to insulin. On the other side, insulin might interfere with MIF action as well, acting as an anti-inflammatory mediator. Therefore, the proper interaction between MIF and insulin is crucial for maintaining homeostasis, while anti-inflammatory therapies based on the systemic MIF blockage may disturb this balance. This review covers MIF-insulin relationship in the physiological and pathological conditions and discusses the approaches for MIF inhibition and their net effect specifically considering possible impact on insulin misfolding and the possible misinterpretation of previous results due to the discovery of MIF functional homolog D-dopachrome tautomerase

Inner Ear Malformations in Congenital Deafness Are Not Associated with Increased Risk of Breech Presentation.

Background: There is speculation that an immature vestibular system may be associated with breech presentation at delivery. Our aim was to determine whether syndromes with congenital inner ear malformations were accompanied by a higher frequency of breech presentation/malpresentations than in the general population (2%-3%). Methods: A review was conducted for published literature using PubMed/MEDLINE (1936-2016), to determine frequency of breech presentation and transverse lie in cases with congenital deafness (Michel aplasia, Wildervanck syndrome, Mondini-Alexander dysplasia, Waardenburg syndrome, CHARGE syndrome, Large vestibular aqueductal syndrome, Pendred syndrome, Oculo-aurico-vertebral spectrum, Jervel and Lange-Nielsen syndrome, Usher syndrome, and Scheibe dysplasia) and vestibular nerve aplasia. Results: Identified were total of 122 cases. The frequency of breech presentation was 1.64%, and of transverse lie 1.64%, giving a total of 3.28% malpresentations. Conclusion: The results of the study suggest that congenital malformations of the vestibular apparatus are not associated with the increased risk of breech presentation at delivery

Establishing the chromatographic fingerprints of flavan‐3‐ols and proanthocyanidins from rose hip ( Rosa sp.) species

The profile of flavan-3-ols and proanthocyanidins in five different Rosa species (R. canina, R. glutinosa, R. rubiginosa, R. multiflora, and R. spinosissima) was estimated on high performance thin layer chromatography cellulose plates. Differences in flavanol and proanthocyanidin profiles of the extracts were evident, among which Rosa spinosissima stood out with catechin as the only detected flavanol and red zones as indication of anthocyanins. Furthermore, the elution solvent for thin layer chromatography with mass spectrometry analyses of glycosylated flavan-3-ols and proanthocyanidins was optimized, enabling identification of catechin, (epi)catechin hexoside, proanthocyanidin dimer, and proanthocyanidin dimers and trimers hexosides. A total of 15 flavanols and their derivatives were identified using ultra-high-performance liquid chromatography with linear trap quadrupole-Orbitrap mass analyzer and epicatechin, gallocatechin, and proanthocyanidin trimer were identified only using this technique. However, proanthocyanidin trimer trihexoside was identified only by thin-layer chromatography with mass spectrometry. To establish the relationships between the flavanols and proanthocyanidins composition of rose hip and their origin, principal component analysis was performed on the entire set of liquid chromatography/mass spectrometry data. Both principal components’ scores plots showed that Rosa spinosissima could be considered as an outlier. Our study demonstrated that flavanol and proanthocyanidin profiles of different rose hips depend on the geographical origin rather than on the cultivar and genotype

Morphological integration and serial homology: A case study of the cranium and anterior vertebrae in salamanders

Serial homology or the repetition of equivalent developmental units and their derivatives is a phenomenon encountered in a variety of organisms, with the vertebrate axial skeleton as one of the most notable examples. Serially homologous structures can be viewed as an appropriate model system for studying morphological integration and modularity, due to the strong impact of development on their covariation. Here, we explored the pattern of morphological integration of the cranium and the first three serially homologous structures (atlas, first, and second trunk vertebrae) in salamandrid salamanders, using micro‐CT scanning and three‐dimensional geometric morphometrics. We explored the integration between structures at static and evolutionary levels. Effects of allometry on patterns of modularity were also taken into account. At the static level (within species), we analyzed inter‐individual variation in shape to detect functional modules and intra‐individual variation to detect developmental modules. Significant integration (based on inter‐individual variation) among all structures was detected and allometry is shown to be an important integrating factor. The pattern of intra‐individual, asymmetric variation indicates statistically significant developmental integration between the cranium and the atlas and between the first two trunk vertebrae. At the evolutionary level (among species), the cranium, atlas, and trunk vertebrae separate as different modules. Our results show that morphological integration at the evolutionary level coincides with morphological and functional differentiation of the axial skeleton, allowing the more or less independent evolutionary changes of the cranial skeleton and the vertebral column, regardless of the relatively strong integration at the static level. The observed patterns of morphological integration differ across levels, indicating different impacts of developmental and phylogenetic constraints and functional demands