Desmoglein 2 Compensates for Desmoglein 3 but Does Not Control Cell Adhesion via Regulation of p38 Mitogen-activated Protein Kinase in Keratinocytes

Desmosomal cadherins are transmembrane adhesion molecules that provide cell adhesion by interacting in the intercellular space of adjacent cells. In keratinocytes, several desmoglein (Dsg1–4) and desmocollin (Dsc1–3) isoforms are coexpressed. We have shown previously that Dsg2 is less important for keratinocyte cohesion compared with Dsg3 and that the latter forms a complex with p38 MAPK. In this study, we compared the involvement of Dsg2 and Dsg3 in the p38 MAPK-dependent regulation of keratinocyte cohesion. We show that loss of cell adhesion and keratin filament retraction induced by Dsg3 depletion is ameliorated by specific p38 MAPK inhibition. Furthermore, in contrast to depletion of Dsg2, siRNA-mediated silencing of Dsg3 induced p38 MAPK activation, which is in line with immunoprecipitation experiments demonstrating the interaction of activated p38 MAPK with Dsg3 but not with Dsg2. Cell fractionation into a cytoskeleton-unbound and a cytoskeleton-anchored desmosome-containing pool revealed that Dsg3, in contrast to Dsg2, is present in relevant amounts in the unbound pool in which activated p38 MAPK is predominantly detectable. Moreover, because loss of cell adhesion by Dsg3 depletion was partially rescued by p38 MAPK inhibition, we conclude that, besides its function as an adhesion molecule, Dsg3 is strengthening cell cohesion via modulation of p38 MAPK-dependent keratin filament reorganization. Nevertheless, because subsequent targeting of Dsg3 in Dsg2-depleted cells led to drastically enhanced keratinocyte dissociation and Dsg2 was enhanced at the membrane in Dsg3 knockout cells, we conclude that Dsg2 compensates for Dsg3 loss of function.     

First evidence of Wolbachia infection in populations of grasshopper Podisma sapporensis (Orthoptera: Acrididae)

The brachypterous grasshopper Podisma sapporensis (Orthoptera: Acrididae) is distributed throughout the Sakhalin, Kunashir and Hokkaido Islands. Karyotypes of this species consist of two major chromosomal races with different sex chromosome systems, XO/XX and XY/XX. Molecular phylogeographic analysis of the chromosome races and subraces confirms the genetic divergence of the races and subraces in P. sapporensis. Here we first report that P. sapporensis is infected with Wolbachia consisting of three variants on wsp locus, while gatB locus was monomorphic. Furthermore, observation of cell tissue of P. sapporensis using electron microscopy confirmed the infection of Wolbachia that was inferred from polymerase chain reaction and revealed the distribution of the bacteria in the head, thorax and abdomen of P. sapporensis embryos. Our finding may shed new light on Wolbachia as a possible agent causing hybrid dysfunction resulting from experimental crosses between chromosome races or subraces of P. sapporensis.     

Scale- and taxon-dependent patterns of plant diversity in steppes of Khakassia,South Siberia (Russia)

The drivers of plant richness at fine spatial scales in steppe ecosystems are still not sufficiently understood. Our main research questions were: (i) How rich in plant species are the natural steppes of Southern Siberia compared to natural and semi-natural grasslands in other regions of the Palaearctic? (ii) What are the main environmental drivers of the diversity patterns in these steppes? (iii) What are the diversity–environment relationships and do they vary between spatial scales and among different taxonomic groups? We sampled the steppe vegetation (vascular plants, bryophytes and lichens) in Khakassia (Russia) with 39 nested-plot series (0.0001–100-m² plot size) and 54 additional 10-m² quadrats across the regional range of steppe types and measured various environmental variables. We measured β-diversity using z-values of power-law species–area relationships. GLM analyses were performed to assess the importance of environmental variables as predictors of species richness and z-value. Khakassian steppes showed both high α- and β-diversity. We found significant scale dependence for the z-values, which had their highest values at small spatial scales and then decreased exponentially. Total species richness was controlled predominantly by heat load index, mean annual precipitation, humus content and soil skeleton content. The positive role of soil pH was evident only for vascular plant species richness. Similar to other studies, we found that the importance of environmental factors strongly differed among taxonomic groups and across spatial scales, thus highlighting the need to study more than one taxon and more than one plot size to get a reliable picture.     

Crosslinking of Chitosan with Dialdehyde Derivatives of Nucleosides and Nucleotides. Mechanism and Comparison with Glutaraldehyde

In medical and pharmaceutical applications, chitosan is used as a component of hydrogels–macromolecular networks swollen in water. Chemical hydrogels are formed by covalent links between the crosslinking reagents and amino functionalities of chitosan. To date, the most commonly used chitosan crosslinkers are dialdehydes, such as glutaraldehyde (GA). We have developed novel GA like crosslinkers with additional functional groups–dialdehyde derivatives of uridine (oUrd) and nucleotides (oUMP and oAMP)–leading to chitosan-based biomaterials with new properties. The process of chitosan crosslinking was investigated in details and compared to crosslinking with GA. The rates of crosslinking with oUMP, oAMP, and GA were essentially the same, though much higher than in the case of oUrd. The remarkable difference in the crosslinking properties of nucleoside and nucleotide dialdehydes can be clearly attributed to the presence of the phosphate group in nucleotides that participates in the gelation process through ionic interactions with the amino groups of chitosan. Using NMR spectroscopy, we have not observed the formation of aldimine bonds. It can be concluded that the real number of crosslinks needed to cause gelation of chitosan chains may be less than 1%.     

Comparison of Effects of Potassium Iodide and Iodosalicylates on the Antioxidant Potential and Iodine Accumulation in Young Tomato Plants

Iodine (I) is classified as a beneficial element for plants. Until now, there have been only hypotheses regarding the uptakes of organic iodine compounds by plant roots. The purpose of our research was to compare the uptakes and effects of the application of the following mineral and organic iodine compounds on young tomato plants: KI, 5-iodosalicylic (5-ISA), and 3,5-diiodosalicylic (3,5-diISA) acids. An additional control combination included the treatment with salicylic acid (SA) alone. All compounds were introduced into the nutrient solution in 5, 10, 25, and 50 μM I concentrations. It was established that after the application of 5-ISA and 3,5-diISA, iodine is taken up to a smaller extent than from KI. The tested KI, 3,5-diISA, and 5-ISA doses had no negative impact on the growth and development of plants, apart from the reduction of shoot biomass after the application of 3,5-diISA in 10 and 25 µM I doses. All applied compounds, except for SA, caused a reduction of ascorbic acid (AA) content and increase of dehydroascorbic acid (DHA) content in leaves. A significant increase of APX activity was noted only for the highest doses of KI and 5-ISA. None of the iodine compounds, in most tested doses, have substantially increased the CAT and POX activities in tomato leaves. Application of KI decreased the levels of all analyzed sugars in tomato leaves. The effect of iodosalicylates on sugar content varied depending on the compound: when applied in the highest dose 5-ISA increased, while 3,5-diISA decreased the sugar accumulation in tomato plants. In all tested treatments, a reduction of SA content in leaves was noted. We conclude that organic iodine compounds, i.e., 3,5-diISA and 5-ISA, can be taken up by the roots of tomato plants at an early stage of development.

     

The effect of the trabecular microstructure on the pullout strength of suture anchors

This study investigates how the microstructural properties of trabecular bone affect suture anchor performance. Seven fresh-frozen humeri were tested for pullout strength with a 5 mm Arthrex Corkscrew in the greater tuberosity, lesser tuberosity, and humeral head. Micro-computed tomography analysis was performed in the three regions of interest directly adjacent to individual pullout experiments. The morphometric properties of bone mineral density (BMD), structural model index (SMI), trabecular thickness (TbTh), trabecular spacing (TbS), trabecular number (TbN), and connectivity density were compared against suture anchor pullout strength. BMD (r=0.64), SMI (r=?0.81), and TbTh (r=0.71) showed linear correlations to the pullout strength of the suture anchor with p-values<0.0001. A predictive model was developed to explain the variances in the individual BMD, SMI, and TbTh correlations. The multi-variant model of pullout strength showed a stronger relationship (r=0.86) compared to the individual experimental results. This study helps confirm BMD is a major influence on the pullout strength of suture anchors, but also illustrates the importance of local microstructure in pullout resistance of suture anchors.     

Neuronal c-Jun is required for successful axonal regeneration, but the effects of phosphorylation of its N-terminus are moderate

Although neural c-Jun is essential for successful peripheral nerve regeneration, the cellular basis of this effect and the impact of c-Jun activation are incompletely understood. In the current study, we explored the effects of neuron-selective c-Jun deletion, substitution of serine 63 and 73 phosphoacceptor sites with non-phosphorylatable alanine, and deletion of Jun N-terminal kinases 1, 2 and 3 in mouse facial nerve regeneration. Removal of the floxed c-jun gene in facial motoneurons using cre recombinase under control of a neuron-specific synapsin promoter (junΔS) abolished basal and injury-induced neuronal c-Jun immunoreactivity, as well as most of the molecular responses following facial axotomy. Absence of neuronal Jun reduced the speed of axonal regeneration following crush, and prevented most cut axons from reconnecting to their target, significantly reducing functional recovery. Despite blocking cell death, this was associated with a large number of shrunken neurons. Finally, junΔS mutants also had diminished astrocyte and microglial activation and T-cell influx, suggesting that these non-neuronal responses depend on the release of Jun-dependent signals from neighboring injured motoneurons. The effects of substituting serine 63 and 73 phosphoacceptor sites (junAA), or of global deletion of individual kinases responsible for N-terminal c-Jun phosphorylation were mild. junAA mutants showed decrease in neuronal cell size, a moderate reduction in post-axotomy CD44 levels and slightly increased astrogliosis. Deletion of Jun N-terminal kinase (JNK)1 or JNK3 showed delayed functional recovery; deletion of JNK3 also interfered with T-cell influx, and reduced CD44 levels. Deletion of JNK2 had no effect. Thus, neuronal c-Jun is needed in regeneration, but JNK phosphorylation of the N-terminus mostly appears to not be required for its function.     

Interaction and functional cooperation between the serine/threonine kinase bone morphogenetic protein type II receptor with the tyrosine kinase stem cell factor receptor

Transmembrane receptors with intrinsic serine/threonine or tyrosine kinase domains regulate vital functions of cells in multicellular eukaryotes, e.g., differentiation, apoptosis, and proliferation. Here, we show that bone morphogenetic protein type II receptor (BMPR-II) which has a serine/threonine kinase domain, and stem cell factor receptor (c-kit) which contains a tyrosine kinase domain form a complex in vitro and in vivo; the interaction is induced upon treatment of cells with BMP2 and SCF. Stem cell factor (SCF) modulated BMP2-dependent activation of Smad1/5/8 and phosphorylation of Erk kinase. SCF also enhanced BMP2-dependent differentiation of C2C12 cells. We found that BMPR-II was phosphorylated at Ser757 upon co-expression with and activation of c-kit. BMPR-II phosphorylation required intact kinase activity of BMPR-II. Abrogation of the c-kit/SCF-dependent phosphorylation of BMPR-II at the Ser757 interfered with the cooperative effect of BMP2 and SCF. Our data suggest that the complex formation between c-kit and BMPR-II leads to phosphorylation of BMPR-II at Ser757, which modulates BMPR-II-dependent signaling.