Influence of tea tree oil (Melaleuca alternifolia) on the cattle tick Rhipicephalus microplus

The aim of this study was to verify the influence of tea tree oil (TTO) (Melaleuca alternifolia) tested in its pure and nanostructured (TTO nanoparticles) forms on the reproduction of female Rhipicephalus microplus. For our purpose, female ticks were collected from naturally infected animals and treated in vitro with TTO (1, 5, and 10 %) and TTO nanoparticles (0.075, 0.375, and 0.75 %). In order to validate the tests, they were performed in triplicate using positive (amitraz) and negative (untreated) controls. It was possible to observe that pure TTO (5 and 10 %) and TTO nanoparticles (0.375 and 0.75 %) showed 100 % reproductive inhibition on female ticks. Additionally, pure TTO (1 %) also showed an acaricide effect (70 %), similarly to the positive control (78.3 %). This is the first study demonstrating the activity of pure TTO and TTO nanoparticles on female ticks. Therefore, based on these results, we were able to show that both forms and all concentrations of M. alternifolia affected tick reproduction by inhibiting egg laying and hatching. We were also able to show that TTO nanoparticles potentiated the inhibitor effect of pure TTO on the reproduction of R. microplus.     

Interim Response of Wading Birds (Pelecaniformes and Ciconiiformes) and Waterfowl (Anseriformes) to the Kissimmee River Restoration Project,Florida, U.S.A

Success of the Kissimmee River Restoration Project will be evaluated in part by monitoring populations of wading birds (Pelecaniformes and Ciconiiformes) and waterfowl (Anseriformes). These two waterbird guilds were integral components of the pre‐channelization river–floodplain ecosystem, and both declined substantially following channelization. Restoration is expected to attract wading birds and waterfowl by reintroducing naturally fluctuating water levels, seasonal hydroperiods, and historic vegetation communities. Post‐construction aerial surveys (November 2001 to May 2008) within the Phase I restoration area indicate that the abundance and species richness of both wading birds and waterfowl have shown a positive restoration response thus far. Dry season abundance of aquatic wading birds and waterfowl has exceeded restoration expectations (≥30.6 birds/km² and ≥3.9 birds/km², respectively) each year since the completion of restoration Phase I in 2001. While there has been a significant positive restoration effect on waterfowl abundance, waterfowl species richness (n = 6) has not yet reached the restoration expectation of ≥13 species. Abundance of the terrestrial cattle egret (Bubulcus ibis), which increased dramatically after the majority of floodplain wetlands were converted to cattle pastures in the channelized system, has shown a significant negative response to restoration. It is anticipated that completion of the remaining phases of restoration (II/III), and implementation of the Kissimmee River Headwaters Revitalization water regulation schedule by 2019, will further increase and improve habitat for wading birds and waterfowl by reestablishing floodplain hydrology that more closely mimics historical conditions.     

L78部分氨基酸残基对SERCA1a钙转运功能的影响

跨肌浆网膜的钙离子ATP酶SERCA1a可在水解ATP的同时逆浓度梯度从胞浆转运钙离子入肌浆网,引发收缩的骨骼肌细胞舒张.目前对SERCA1a结构与功能的研究,要领先于对其他P型离子转运ATP酶的研究.为了解SERCA1a跨膜螺旋M7与M8膜内侧连接部Linker78(L78)的功能,我们评估了L78部分氨基酸残基突变对SERCA1a反应循环的影响,发现:除G864A之外的所有突变体均可不同程度地降低钙离子转运速率;G862或P863氨基酸残基的突变导致SERCA1a的ATP酶活性显著降低或丧失,并引起由ATP或无机磷酸Pi生成的磷酸化酶中间体EP量显著减少;A893被P取代对SERCA1a的影响与G862、P863突变相似;与野生型SERCA1a相比,G864A与FMQ873-875单突变体ATP酶活性及EP生成量无明显差别.上述结果表明,M7、M8膜内侧连接部L78不仅与跨膜区钙离子转运过程密切相关,而且L78在GPG862-864处及A893附近的正确转折对胞浆结构域P的磷酸化也具有关键的远距离调控作用,提示L78的结构及柔度对SERCA1a构象正常的周期性变化至关重要.     

Polymorphisms of vitamin D receptor gene in the population of eastern Croatia with psoriasis vulgaris and diabetes mellitus

The aim of this study was to evaluate the possible association between polymorphisms in the Vitamin D receptor gene (VDR gene) and tendency for development of psoriasis vulgaris and diabetes mellitus in the population of Slavonia, which is a region in the Eastern Croatia. In order to conduct the mentioned evaluation the restriction fragment length polymorphisms (ApaI, BsmI and TaqI) in the Vitamin D receptor gene were researched in three groups of patients: patients suffering only from psoriasis vulgaris, patients suffering only from diabetes mellitus, and patients suffering at the same time from both diseases. Four most common genotypes were found in all standardized control patients: triple heterozygotes BbAaTt (in 29.3% of the studied patients), bbAaTT (in 18.6% of the studied patients), bbaaTT (in 12.9% of the studied patients) and BbAATt (in 8.6% of the studied patients). Three most common VDR 3'-RFLP haplotypes determined in this study were: three-component baT, Bat and bAT haplotype. Results of the Hardy-Weinberg equilibrium showed presence of BsmI polymorphism genotype frequencies disequilibrium in the group of patients suffering from psoriasis and ApaI polymorphism in the group of patients suffering from both diseases. According to the same statistical test all conditions for TaqI polymorphism genotype frequency were fulfilled in all groups of studied patients. There was no significant difference in distribution of BsmI, ApaI or TaqI polymorphism genotype frequencies between control patients and any of the subgroup of studied patients. In studied population none of analysed polymorphisms individually was associated with the risk of development of psoriasis, diabetes or combined phenotype.     

Molecular phylogeny of the Robust clade (Faviidae, Mussidae, Merulinidae, and Pectiniidae): An Indian Ocean perspective

Recent phylogenetic analyses have demonstrated the limits of traditional coral taxonomy based solely on skeletal morphology. In this phylogenetic context, Faviidae and Mussidae are ecologically dominant families comprising one third of scleractinian reef coral genera, but their phylogenies remain partially unresolved. Many of their taxa are scattered throughout most of the clades of the Robust group, and major systematic incongruences exist. Numerous genera and species remain unstudied, and the entire biogeographic area of the Indian Ocean remains largely unsampled. In this study, we analyzed a portion of the mitochondrial cytochrome c oxidase subunit 1 gene and a portion of ribosomal DNA for 14 genera and 27 species of the Faviidae and Mussidae collected from the Indian Ocean and New Caledonia and this is the first analysis of five of these species. For some taxa, newly discovered evolutionary relationships were detected, such as the evolutionary distinctiveness of Acanthastrea maxima, the genetic overlap of Parasimplastrea omanensis and Blastomussa merleti, and the peculiar position of Favites peresi in clade XVII together with Echinopora and Montastraea salebrosa. Moreover, numerous cases of intraspecific divergences between Indian Ocean and Pacific Ocean populations were detected. The most striking cases involve the genera Favites and Favia, and in particular Favites complanata, F. halicora, Favia favus, F. pallida, F. matthaii, and F. rotumana, but divergence also is evident in Blastomussa merleti, Cyphastrea serailia, and Echinopora gemmacea. High morphological variability characterizes most of these taxa, thus traditional skeletal characteristics, such as corallite arrangement, seem to be evolutionary misleading and are plagued by convergence. Our results indicate that the systematics of the Faviidae and the Mussidae is far from being resolved and that the inclusion of conspecific populations of different geographical origin represents an unavoidable step when redescribing the taxonomy and systematics of scleractinian corals. More molecular phylogenies are needed to define the evolutionary lineages that could be corroborated by known and newly discovered micromorphological characters.     

Visualization and quantification of cardiac mitochondrial protein clusters with STED microscopy

The visualization and quantification of mitochondria-associated proteins with high power microscopy methods is of particular interest to investigate protein architecture in this organelle. We report the usage of a custom-made STimulated Emission Depletion (STED) fluorescence nanoscope with ~30nm lateral resolution for protein mapping of Percoll-purified viable mitochondria from murine heart. Using this approach, we were able to quantify and resolve distinct protein clusters within mitochondria; specifically, cytochrome c oxidase subunit 2 is distributed in clusters of ~28nm; whereas the voltage dependent anion channel 1 displays three size distributions of ~33, ~55 and ~83nm.     

Differentiation Increases the Resistance of Neuronal Cells to Amyloid Toxicity

A substantial lack of information is recognized on the features underlying the variable susceptibility to amyloid aggregate toxicity of cells with different phenotypes. Recently, we showed that different cell types are variously affected by early aggregates of a prokaryotic hydrogenase domain (HypF-N). In the present study we investigated whether differentiation affects cell susceptibility to amyloid injury using a human neurotypic SH-SY5Y cell differentiation model. We found that retinoic acid-differentiated cells were significantly more resistant against Aβ1-40, Aβ1-42 and HypF-N prefibrillar aggregate toxicity respect to undifferentiated cells treated similarly. Earlier and sharper increases in cytosolic Ca²⁺ and ROS with marked lipid peroxidation and mitochondrial dysfunction were also detected in exposed undifferentiated cells resulting in apoptosis activation. The reduced vulnerability of differentiated cells matched a more efficient Ca²⁺-ATPase equipment and a higher total antioxidant capacity. Finally, increasing the content of membrane cholesterol resulted in a remarkable reduction of vulnerability and ability to bind the aggregates in either undifferentiated and differentiated cells. Special issue article in honor of Dr. Anna Maria Giuffrida-Stella.     

Genotoxicity of aminohydroxynaphthoquinones in bacteria, yeast, and Chinese hamster lung fibroblast cells

There are few studies on the biological activity of aminohydroxy derivates of 1,4-naphthoquinone (1,4-NQ) on prokaryotic and eukaryotic cells. We determined the mutagenic activity of 5-amino-8-hydroxy-1,4-naphthoquinone (ANQ) and 5-amino-2,8-dihydroxy-1,4-naphthoquinone (ANQ-OH) as compared to the unsubstituted 1,4-NQ in Salmonella/microsome assay. Potential mutagenic and recombinogenic effects and cytotoxicity were analyzed in haploid and diploid cultures of the yeast Saccharomyces cerevisiae. In Salmonella/microsome assay, 1,4-NQ was not mutagenic, whereas aminohydroxynaphthoquinones were weakly mutagenic in TA98 and TA102 strains. In haploid yeast in stationary growth phase (STAT), mutagenic response was only observed for the hom3 locus at the highest dose. In diploid yeast, aminohydroxynaphthoquinones did not induce any recombinogenic events, but 1,4-NQ was shown to be a recombinogenic agent. These results suggest that aminohydroxynaphthoquinones are weak mutagenic agents only in prokaryotic cells. The cytotoxicity of 1,4-NQ in yeast stationary cells was more significant in diploid cells as compared to that observed in haploid cells. However, ANQ and ANQOH were slightly cytotoxic in all treatments. Genotoxicity of these naphthoquinone compounds was also determined in V79 Chinese hamster lung fibroblast cells using standard Comet, as well as modified Comet assay with the bacterial enzymes formamidopyrimidine DNA-glycosylase (FPG) and endonuclease III (ENDOIII). Both 1,4-NQ and ANQ induced pronounced DNA damage in the standard Comet assay. The genotoxic effect of ANQ-OH was observed only at the highest dose. In presence of metabolic activation all substances showed genotoxic effects on V79 cells. Post-treatment of V79 cells with ENDOIII and FPG proteins did not have a significant effect on ANQ-OH-induced oxidative DNA damage as compared to standard alkaline Comet assay. However, all naphthoquinones were genotoxic in V79 cells in the presence of metabolic activation and post-treatment with enzymes, indicating that all compounds induced oxidative DNA damage in V79 cells. Our data suggest that aminohydroxynaphthoquinone pro-oxidant activity, together with their capability of DNA intercalation, have an important role in mutagenic and genotoxic activities.