Formation of lanthanide(III) texaphyrin complexes with DNA controlled by the size of the central metal cation

Interaction of lanthanide(III) texaphyrins (LnTEX) with calf thymus DNA was studied by UV-Vis, CD, resonance light scattering spectroscopy and by laser flash photolysis. The thermal stability of the LnTEX/DNA complexes decreases with the increasing lanthanide ion radius: LuTEX>DyTEX>GdTEX>EuTEX>CeTEX> or =LaTEX. Texaphyrins LaTEX, LuTEX and GdTEX produce O2(1Deltag) in methanol solutions. In a phosphate buffer, the concentration of O2(1Deltag) produced by these texaphyrins bound to DNA is affected by the formation of aggregates on the DNA backbone.     

Ultrastructure, development, and homology of insect embryonic cuticles

Ultrastructure and deposition of the cuticles secreted by embryos representing eight insect orders were examined by transmission and scanning electron microscopy. Embryos of the apterygote silverfish Thermobia domestica deposit two embryonic cuticles. Deposition of the first (EC1) is initiated at the beginning of appendage development when the intercalary segment and the neural groove are clearly visible. This cuticle lacks surface microsculpture and consists of an outer epicuticle and an underlying fibrous layer, thought to represent procuticle. At the time of dorsal closure, deposition of a second embryonic cuticle (EC2) begins; this bears sensilla and functions in the first instar larva. In representative embryos of seven pterygote orders (Ephemeroptera, Odonata, Plecoptera, Neuroptera, Coleoptera, Lepidoptera, and Mecoptera), three cuticles were found to be secreted. The first cuticle in pterygotes is homologous to EC1 of T. domestica, but consists solely of outer epicuticle. EC2, the "prolarval cuticle," bears a characteristic surface microsculpture in embryos of some species and egg-teeth and other hatching devices, and consists of outer and inner epicuticles and a more or less reduced procuticle. EC2 is reduced in the embryos of derived endopterygotes, where a procuticle is lacking and the inner epicuticle is reduced. After hatching, when EC2 is shed, the first instar larva is covered by a third embryonic cuticle (EC3), whose deposition was initiated while the insect was still within the egg. Presence of only two embryonic cuticles in cyclorrhaphous flies is due to the total loss of prolarval cuticle. Investigated exopterygote and endopterygote insects excluding flies thus deposit three embryonic cuticles, and their juveniles (exopterygote "nymphs"; endopterygote "larvae") seem to hatch at equivalent stages of development. Differences between the modes of cuticulogenesis in silverfish and pterygote embryos suggest that the apterygote first larval instar was embryonized and became a fully embryonic prolarva in pterygotes.     

Role of apoptosis and proliferation in differentiation of human retina,regio olfactoria and regio respiratoria

Apoptosis and proliferation are very important processes during central nervous system development. In our study we concentrated on human fetus retina, regio respiratoria and olfactoria. The aims of our work were to evaluate a proliferative activity proved by PCNA and KI-67 gene expressions, to measure an expression of anti-apoptotic gene bcl-2, and finally to compare levels of apoptosis and proliferation in the areas investigated. Altogether we studied 29 fetuses between the 5th to 13th week of gestation (WG). Parallel sections were elaborated by using standard methods of immunohistochemistry for the detection of proteins PCNA, Bcl-2 and Ki-67. Apoptotic cells were detected by two methods: TUNEL (for the detection of DNA fragmentation) and Apostain (for evidence of ssDNA). The results of semiquantitative evaluation show that expressions of proteins PCNA and KI-67 begins after the 6th WG, from the 8th to 13th WG they are very high in all investigated areas. Occurrence of apoptotic cells in regio respiratoria is sporadic between the 5th to 9th WG, while later it is widely spread. Apoptosis in regio olfactoria appears after the 9th WG and it is also very extensive. In the retina we found a higher frequency of apoptotic cells only in the 11th to 13th WG. The results of both methods for detection of apoptosis were concordant. Anti-apoptotic gene bcl-2 was detectable in all examined areas, but it followed no orderly pattern. We conclude, that the expression of apoptosis increases with the age from the 7th to 13th WG, and that both apoptosis and proliferation participate in the morphogenesis of fetuses between the 5th to 13th WG.     

Analysis of binding of mannosides in relation to Langerin (CD207) in Langerhans cells of normal and transformed epithelia

Tandem-repeat C-type lectins (pattern-recognition receptors) with specificity for mannosides are intimately involved in antigen recognition, uptake, routing and presentation in macrophages and dendritic cells. In Langerhans cells, Langerin (CD207), a type-II transmembrane protein with a single C-type carbohydrate recognition domain attached to a heptad repeat in the neck region, which is likely to establish oligomers with an -coiled-coil stalk, has been implicated in endocytosis and the formation of Birbeck granules. The structure of Langerin harbours essential motifs for Ca²⁺-binding and sugar accommodation. Lectin activity has previously been inferred by diminished antibody binding to cells in the presence of the glycan ligand mannan. In view of the complexity of the C-type lectin/lectin-like network, it is unclear what role Langerin plays for Langerhans cells in binding mannosides. In order to reveal in frozen tissue sections to what extent mannose-binding activity co-localizes with Langerin, we have used a synthetic marker, i.e. a neoglycoprotein carrying mannose maxiclusters, as a histochemical ligand, and computer-assisted fluorescence monitoring in a double-labelling procedure. Mannoside-binding capacity was detected in normal epithelial cells. Double labelling ensured the unambiguous assessment of the binding of the neoglycoprotein in Langerhans cells. Light-microscopically, its localization profile resembled the pattern of immunohistochemical detection of Langerin. This result has implications for suggesting rigorous controls in histochemical analysis of this cell type, because binding of kit reagents, i.e. mannose-rich glycoproteins horseradish peroxidase or avidin, to Langerin (or a spatially closely associated lectin) could yield false-positive signals. To show that recognition of carbohydrate ligands in dendritic cells is not restricted to mannose clusters, we have also documented binding of carrier-immobilized histo-blood group A trisaccharide, a ligand of galectin-3, which was not affected by the presence of a blocking antibody to Langerin. Remarkably, access to the carbohydrate recognition domain of Langerin appeared to be impaired in proliferatively active environments (malignancies, hair follicles), indicating presence of an endogenous ligand with high affinity to saturate the C-type lectin under these conditions.     

Google Trends reflect allergic rhinitis symptoms related to birch and grass pollen seasons

Google Trends (GT) describes the variation of the relevant interest of internet searches toward medical conditions and related symptoms. Allergic rhinitis symptom levels result from the intensity of exposure to aeroallergens in combination with relevant medication use. We analyze data from Germany to examine the relationship between hay fever-related Google search terms, symptom levels, medication use, and pollen count levels. For doing so, we also employ the new definitions on pollen season and peak pollen period start and end as proposed by the European Academy of Allergy and Clinical Immunology in a recently published position paper. We extract GT data for a number of search terms related to allergic rhinitis for Germany. We use total nasal symptom and mediation scores as reported by patients via a patient hay fever diary in the Berlin and Brandenburg areas in Germany for 3 years (2014–2016), accompanied by pollen data. Then a Pearson and Spearman correlation analysis is performed between symptom data and GT data. A graphical analysis is conducted, and the identification of pollen season and peak pollen periods is done based on the EAACI criteria. The analysis reveals that GT data are highly correlated with symptom levels and follow peak pollen period start–end, concerning grass and birch pollen-induced allergic rhinitis symptoms. GT data can be used as a proxy for the identification of the onset and variation of nasal symptom and medication score for allergic rhinitis sufferers.     

Rapid Identification of Medically Important Candida Isolates Using High Resolution Melting Analysis

An increasing trend in non albicans infections and various susceptibility patterns to antifungal agents implies a requirement for the quick and reliable identification of a number of medically important Candida species. Real-time PCR followed by high resolution melting analysis (HRMA) was developed, tested on 25 reference Candida collection strains and validated on an additional 143 clinical isolates in this study. All reference strains and clinical isolates inconclusive when using phenotypic methods and/or HRMA were analysed using ITS2 sequencing. Considering reference and clinical strains together, 23 out of 27 Candida species could be clearly distinguished by HRMA, while the remaining 4 species were grouped in 2 pairs, when applying the mean Tm ± 3 SD values, the shape of the derivative melting curve (dMelt curve) and, in some cases, the normalized and temperature—shifted difference plot against C. krusei. HRMA as a simple, rapid and inexpensive tool was shown to be useful in identifying a wide spectrum of clinically important Candida species. It may complement the current clinical diagnostic approach based on commercially available biochemical kits.     

New insights into Legionella pneumophila biofilm regulation by c-di-GMP signaling

The waterborne pathogen Legionella pneumophila grows as a biofilm, freely or inside amoebae. Cyclic-di-GMP (c-di-GMP), a bacterial second messenger frequently implicated in biofilm formation, is synthesized and degraded by diguanylate cyclases (DGCs) and phosphodiesterases (PDEs), respectively. To characterize the c-di-GMP-metabolizing enzymes involved in L. pneumophila biofilm regulation, the consequences on biofilm formation and the c-di-GMP concentration of each corresponding gene inactivation were assessed in the Lens strain. The results showed that one DGC and two PDEs enhance different aspects of biofilm formation, while two proteins with dual activity (DGC/PDE) inhibit biofilm growth. Surprisingly, only two mutants exhibited a change in global c-di-GMP concentration. This study highlights that specific c-di-GMP pathways control L. pneumophila biofilm formation, most likely via temporary and/or local modulation of c-di-GMP concentration. Furthermore, Lpl1054 DGC is required to enable the formation a dense biofilm in response to nitric oxide, a signal for biofilm dispersion in many other species.