Candidate gene studies of diabetic retinopathy in human

Diabetic retinopathy (DR) is a multifactorial disease with complex pathophysiology. It is the main cause of blindness among the people in productive age. The purpose of this literature review is to highlight recent achievements in the genetics of diabetic retinopathy with particular focus on candidate gene studies. We summarized most of the available published data about candidate genes for diabetic retinopathy with the goal to identify main genetic aspects. We conclude that genetic studies reported contradictory findings and no genetic variants meet criteria of a diagnostic marker, or significantly elucidate the root of DR development. Based on these findings it is important to continue with the research in the field of DR genetics, mainly due to the fact that currently new possibilities and approaches associated with utilization of next-generation sequencing are available.     

Effects of Dietary Phosphate on Adynamic Bone Disease in Rats with Chronic Kidney Disease – Role of Sclerostin?

High phosphate intake is known to aggravate renal osteodystrophy along various pathogenetic pathways. Recent studies have raised the possibility that dysregulation of the osteocyte Wnt/β-catenin signaling pathway is also involved in chronic kidney disease (CKD)-related bone disease. We investigated the role of dietary phosphate and its possible interaction with this pathway in an experimental model of adynamic bone disease (ABD) in association with CKD and hypoparathyroidism. Partial nephrectomy (Nx) and total parathyroidectomy (PTx) were performed in male Wistar rats. Control rats with normal kidney and parathyroid function underwent sham operations. Rats were divided into three groups and underwent pair-feeding for 8 weeks with diets containing either 0.6% or 1.2% phosphate: sham 0.6%, Nx+PTx 0.6%, and Nx+PTx 1.2%. In the two Nx+PTx groups, serum creatinine increased and blood ionized calcium decreased compared with sham control group. They also presented hyperphosphatemia and reduced serum parathyroid hormone (PTH) and fibroblast growth factor 23 (FGF23) levels. Fractional urinary excretion of phosphate increased in Nx+PTx 1.2% rats despite lower PTH and FGF23 levels than in sham group. These biochemical changes were accompanied by a decrease in bone formation rates. The Nx+PTx 1.2% group had lower bone volume (BV/TV), higher osteoblast and osteocyte apoptosis, and higher SOST and Dickkopf-1 gene expression than the Nx+PTx 0.6% group. Nx+PTx 0.6% rat had very low serum sclerostin levels, and Nx+PTx 1.2% had intermediate sclerostin levels compared with sham group. Finally, there was a negative correlation between BV/TV and serum sclerostin. These results suggest that high dietary phosphate intake decreases bone volume in an experimental model of CKD-ABD, possibly via changes in SOST expression through a PTH-independent mechanism. These findings could have relevance for the clinical setting of CKD-ABD in patients who low turnover bone disease might be attenuated by optimal control of phosphate intake and/or absorption.     

The use of esterase polymorphism for analysis of the genetic diversity and structure of stevia (Stevia rebaudiana Bert. Bertoni) populations

Native polyacrylamide gel electrophoresis was used in the current study to identify polymorphisms in α- and β-esterase loci in the leaves of Stevia rebaudiana Bert. Bertoni. The esterases produced from the Est-2 and Est-4 loci were observed as strongly stained bands, and four alleles were detected at each of these loci from the 428 analysed S. rebaudiana plants. The observed and expected mean heterozygosity were higher in populations maintained by genetically breeding plants for higher glycoside production than in plant populations propagated by seeds or maintained by vegetative propagation. The esterase analysis employed in this study showed that the artificial selection of plant samples with specific uniform characteristics, such increased height and precocious flowering, may lead to the fixation of alleles and α/β-esterase phenotype patterns in S. rebaudiana populations. The uniform α/β-esterase phenotype may be applied in the monitoring of genetic stability in selected populations for specific traits of interest.     

Effects of single and combined cell treatments based on low pH and high concentrations of ethanol on the growth and fermentation of Dekkera bruxellensis and Saccharomyces cerevisiae

The alcoholic fermentation in Brazil displays some peculiarities because the yeast used is recycled in a non-aseptic process. After centrifugation, the cells are treated with acid to control the bacterial growth. However, it is difficult to manage the indigenous yeasts without affecting the main culture of Saccharomyces cerevisiae. This work evaluated how the cell treatment could be modified to combat contaminant yeasts based on the differential sensitivities to low pH and high concentrations of ethanol displayed by an industrial strain of S. cerevisiae and three strains of Dekkera bruxellensis, which are common contaminant yeasts in Brazilian fermentation processes. The tests were initially performed in rich medium with a low pH or a high concentration of ethanol to analyse the yeast growth profile. Then, the single and combined effects of low pH and ethanol concentration on the yeast cell viability were evaluated under non-proliferative conditions. The effects on the fermentation parameters were also verified. S. cerevisiae grew best when not subjected to the stresses, but this yeast and D. bruxellensis had similar growth kinetics when exposed to a low pH or increased ethanol concentrations. However, the combined treatments of low pH (2.0) and ethanol (11 or 13 %) resulted in a decrease of D. bruxellensis cell viability almost three times higher than of S. cerevisiae, which was only slightly affected by all cell treatments. The initial viability of the treated cells was restored within 8 h of growth in sugar cane juice, with the exception of the combined treatment for D. bruxellensis. The ethanol-based cell treatment, in despite of slowing the fermentation, could decrease and maintain D. bruxellensis population under control while S. cerevisiae was taking over the fermentation along six fermentative cycles. These results indicate that it may be possible to control the growth of D. bruxellensis without major effects on S. cerevisiae. The cells could be treated between the fermentation cycles by the parcelled addition of 13 % ethanol to the tanks in which the yeast cream is treated with sulphuric acid at pH 2.0.     

Dysfunction of heterotrimeric kinesin-2 in rod photoreceptor cells and the role of opsin mislocalization in rapid cell death

Due to extensive elaboration of the photoreceptor cilium to form the outer segment, axonemal transport (IFT) in photoreceptors is extraordinarily busy, and retinal degeneration is a component of many ciliopathies. Functional loss of heterotrimeric kinesin-2, a major anterograde IFT motor, causes mislocalized opsin, followed by rapid cell death. Here, we have analyzed the nature of protein mislocalization and the requirements for the death of kinesin-2-mutant rod photoreceptors. Quantitative immuno EM showed that opsin accumulates initially within the inner segment, and then in the plasma membrane. The light-activated movement of arrestin to the outer segment is also impaired, but this defect likely results secondarily from binding to mislocalized opsin. Unlike some other retinal degenerations, neither opsin–arrestin complexes nor photoactivation were necessary for cell loss. In contrast, reduced rod opsin expression provided enhanced rod and cone photoreceptor survival and function, as measured by photoreceptor cell counts, apoptosis assays, and ERG analysis. The cell death incurred by loss of kinesin-2 function was almost completely negated by Rho^−/−. Our results indicate that mislocalization of opsin is a major cause of photoreceptor cell death from kinesin-2 dysfunction and demonstrate the importance of accumulating mislocalized protein per se, rather than specific signaling properties of opsin, stemming from photoactivation or arrestin binding.     

Tanchintongia —eine aberrante permische Bivalve in Europa

Tanchintongia ogulineci n. sp. occurs in the Permian limestone of theNeoschwagerina craticulifera zone of the Velebit mountains (Croatia, Yugoslavia). The shells with a length up to 64 cm are flattened along the flanks, and below are bent to a keel. Instead of the beaks there is an oval opening for the byssus right in front at the top.     

Shifts in the dominance between diatoms and cryptophytes during three late summers in the Bransfield Strait (Antarctic Peninsula)

Recent global warming reduces surface water salinity around the Antarctic Peninsula as a result of the glacial meltwater runoff, which increases the occurrence and abundance of certain phytoplankton groups, such as cryptophytes. The dominance of this particular group over diatoms affects grazers, such as Antarctic krill, which preferentially feed on diatoms. Using three late summer data sets from the Bransfield Strait (2008–2010), we observed variations in the dominant phytoplankton groups determined by HPLC/CHEMTAX pigment analysis and confirmed by microscopy. Results indicate that the dominance of diatoms, particularly in 2008 and 2009, was associated with a deeper upper mixed layer (UML), higher salinity and warmer sea surface temperature. In contrast, cryptophytes, which were dominant in 2010, were associated with a shallower UML, lower salinity and colder sea surface temperatures. The low diatom biomass observed in the summer of 2010 was associated with high nutrient concentration, particularly silicate, and low chlorophyll a (summer monthly average calculated from satellite images). The interannual variability here observed suggests a delayed seasonal succession cycle of phytoplankton in the summer of 2010 associated with a cold summer and a late ice retreat process in the region. This successional delay resulted in a notable decrease of primary producers’ biomass, which is likely to have impacted regional food web interactions. This study demonstrates the susceptibility of the Antarctic phytoplankton community structure to air temperature, which directly influences the timing of ice melting and consequently the magnitude of primary production and succession pattern of phytoplankton groups.     

Light control on phytoplankton production in a shallow and turbid estuarine system

Tagus estuary is one of the largest estuaries of Western Europe. With the aim of unravelling the drivers of primary production in this shallow and turbid nutrient replete estuary, we tested the hypothesis that light availability is a major factor controlling phytoplankton production. Environmental parameters, phytoplankton biomass, community composition, and photosynthetic parameters were monitored at two sites in the estuary during a complete annual cycle. Despite the fact that nutrient concentrations were always above growth-limiting values, Chl a concentrations were relatively low throughout the study period. High water column turbidity, due to riverine inputs, promoted a rapid attenuation of light and created a compressed profile with optimal photosynthetic conditions. Therefore, the phytoplankton community, dominated by small cells, such as diatoms and cryptophycean flagellates, displayed highly photosynthetic efficiency and low light-saturated photosynthetic rates as a photo-acclimation response to low light conditions year-round. Primary production rate was unimodal, peaking in the summer months. In such estuarine system, gross primary production could thus be predicted by an existing robust empirical model based on pigment standing crop (Chl a), surface irradiance (E ₀) and optical depth (Z _eup). Compared to other shallow estuaries, the Tagus can be classified as a low- to moderately productive estuary, being the turbidity-induced low light conditions the principal factor limiting phytoplankton growth.     

Tetra-, penta- and hexacoordinate copper(II) complexes with N3 donor isoindoline-based ligands: Characterization and SOD-like activity

Reaction of 1,3-bis(2′-Ar-imino)isoindolines (HLⁿ, n = 1-7, Ar = benzimidazolyl, N-methylbenzimidazolyl, thiazolyl, pyridyl, 3-methylpyridyl, 4-methylpyridyl, and benzthiazolyl, respectively) with Cu(OCH₃)₂ yields mononuclear hexacoordinate complexes with Cu(Lⁿ)₂ composition. With cupric perchlorate square-pyramidal [Cu^II(HLⁿ)(NCCH₃)(OClO₃)]ClO₄ complexes (n = 1, 3, 4) were isolated as perchlorate salts, whereas with chloride Cu^II(HLⁿ)Cl₂ (n = 1, 4), or square-planar Cu^IICl₂(HLⁿ) (n = 2, 3, 7) complexes are formed. The X-ray crystal structures of Cu(L³)₂, Cu(L⁵)₂, [Cu^II(HL⁴)(NCCH₃)(OClO₃)]ClO₄, Cu^IICl(L²) and Cu^IICl(L⁷) are presented along with electrochemical and spectral (UV-Vis, FT-IR and X-band EPR) characterization for each compound. When combined with base, the isoindoline ligands in the [Cu^II(HLⁿ)(NCCH₃)(OClO₃)]ClO₄ complexes undergo deprotonation in solution that is reversible and induces UV-Vis spectral changes. Equilibrium constants for the dissociation are calculated. X-band EPR measurements in frozen solution show that the geometry of the complexes is similar to the corresponding X-ray crystallographic structures. The superoxide scavenging activity of the compounds determined from the McCord-Fridovich experiment show dependence on structural features and reduction potentials.