Biological evaluation and docking studies of recently identified inhibitors of phosphoinositide-3-kinases

The alpha isoform of the phosphatidylinositol-3-kinases (PI3Kα) is often mutated, amplified and overexpressed in human tumors. In an effort to develop new inhibitors targeting this enzyme, we carried out a pharmacophore model study based on six PI3Kα-selective compounds. The pharmacophore searching identified three structurally novel inhibitors of PI3Kα and its H1047R mutant. Our biological studies show that two of our hit molecules suppressed the formation of pAKT, a downstream effector of PI3Kα, and induced apoptosis in the HCT116 colon cancer cell line. QPLD-based docking showed that residues Asp933, Glu849, Val851, and Gln859 appeared to be key binding residues for active inhibitors.     

Migrating locusts can detect polarized reflections to avoid flying over the sea

The desert locust Schistocerca gregaria is a well known migrating insect, travelling long distances in swarms containing millions of individuals. During November 2004, such a locust swarm reached the northern coast of the Gulf of Aqaba, coming from the Sinai desert towards the southeast. Upon reaching the coast, they avoided flying over the water, and instead flew north along the coast. Only after passing the tip of the gulf did they turn east again. Experiments with tethered locusts showed that they avoided flying over a light-reflecting mirror, and when given a choice of a non-polarizing reflecting surface and a surface that reflected linearly polarized light, they preferred to fly over the former. Our results suggest that locusts can detect the polarized reflections of bodies of water and avoid crossing them; at least when flying at low altitudes, they can therefore avoid flying over these dangerous areas.     

Comparative study of the amount of backflow produced by four types of aortic valve prostheses

To determine the extent of backflow encountered with currently used prosthetic valves, four types of aortic valves with comparable orifice diameters were tested in a pulse duplicating system. These were a Hancock porcine valve, a Lillehei-Kaster pivoting disk valve, a St. Jude bileaflet valve and a Bj?rk-Shiley tilting disk valve. Mean aortic pressure was sequentially increased from 83 to 147 mmHg, keeping the pump rate essentially constant (69-73 strokes/min). The porcine valve produced the least amount of total backflow (backflow due to closure plus leakage backflow) (1.6 to 2.4 mL/stroke). Among the mechanical valves the Bj?rk-Shiley valve showed the least amount of total backflow (5.0 to 6.0 mL/stroke). At a mean aortic pressure of 100 mmHg and a low cardiac output of 2 L/min, the total backflow with the porcine valve was only 6 percent of forward flow; whereas it was 19 percent with the Lillehei-Kaster valve, 22 percent with the St. Jude valve and 18 percent with the Bj?rk-Shiley valve. Leakage backflow at a given level of mean aortic pressure was, as expected, directly related to the annular clearance area. It is concluded that the Hancock valve showed the least amount of backward flow, which would be particularly beneficial in low output states. In the presence of normal hemodynamics, the amount of backflow with the three mechanical valves appeared to be well below the level of backflow considered to be clinically significant.     

Contrasting levels of DNA polymorphism at the autosomal and X-linked visual color pigment loci in humans and squirrel monkeys

The X-linked color pigment (opsin) locus is known to be highly polymorphic in the squirrel monkey and other New World monkeys. To see whether this is also the case for the autosomal (blue) opsin locus, we obtained 32 squirrel monkey and 30 human blue opsin gene sequences. No amino acid polymorphism was found in either the squirrel monkey sample or the human sample, contrary to the situation at the X-linked opsin locus. This sharp contrast in the level of polymorphism might be due to differences in gene expression between the autosomal and the X-linked loci. At the X-linked locus, heterozygote advantage can occur because, owing to X-inactivation, the two alleles in a heterozygote are expressed in different cone cells, producing two types of cone cell, whereas at the autosomal locus, heterozygote advantage cannot occur because the two alleles in a heterozygote are expressed in the same cone cells, producing only one type of cone cell (i.e., phenotypically a homozygote). From the sequence data, the levels of nucleotide diversity (pi, i.e., the number of nucleotide differences per site) are estimated: for the human sample, pi = 0.00% per nondegenerate site, 0.00% per twofold degenerate site, and 0.04% per fourfold degenerate site in the coding regions and 0.01% per site in intron 4; for the squirrel monkey sample, pi = 0.00% per nondegenerate site, 0.00% per twofold degenerate site, and 0.15% per fourfold degenerate site in the coding regions and 0.17% per site in intron 4. The blue opsin genes from the common and pygmy chimpanzees, the gorilla, the capuchin, and the howler monkey were also sequenced. Features critical to the function of the opsin are well conserved in all known mammalian sequences. However, the interhelical loops are, on average, actually more conservative than the transmembrane helical regions. In addition, these sequence data and those from some other genes indicate that the common and pygmy chimpanzees are not closely related, their divergence data being from one third to one half the date of the human-chimpanzee divergence.      

An Arg-Gly-Asp sequence within thrombin promotes endothelial cell adhesion

Thrombin, in addition to its central role in hemostasis, possesses diverse cellular bioregulatory functions implicated in wound healing, inflammation, and atherosclerosis. In the present study we demonstrate that thrombin molecules modified either at the procoagulant or catalytic sites induce endothelial cell (EC) adhesion, spreading, and cytoskeletal reorganization. The most potent adhesive thrombin analogue (NO2-alpha-thrombin) was obtained by nitration of tyrosine residues. The cell adhesion promoting activity of NO2-alpha-thrombin was blocked upon the formation of thrombin-antithrombin III (ATIII) complexes and by antiprothrombin antibodies, but was unaffected by hirudin. Arg-Gly-Asp-containing peptides, fully inhibited EC adhesion to NO2-alpha-thrombin, while synthetic peptides corresponding to thrombin "Loop B" mitogenic site and the thrombin-derived chemotactic fragment "CB67-129", were uneffective. Immunofluorescence studies indicated that EC adhesion to NO2-alpha-thrombin was followed by cell spreading, actin microfilament assembly, and formation of focal contacts. By the use of specific antibodies, the vitronectin (vn) receptor (alpha v beta 3) was found to be localized in clusters upon cell adhesion to NO2-alpha-thrombin. An anti alpha v beta 3 antibody blocked EC adhesion and spreading while antifibronectin (fn) receptor (alpha 5 beta 1) antibodies were uneffective. While native thrombin exhibited a very low cell attachment activity, thrombin that was incubated at 37 degrees C before coating of plastic surfaces induced EC attachment and spreading. We propose that under certain conditions the naturally hindered RGD domain within thrombin is exposed for interaction with alpha v beta 3 on EC. This in turn promotes cell adhesion, spreading, and reorganization of cytoskeletal elements, which may altogether contribute to repair mechanisms in the disturbed vessel wall. This study defines a new biological role of thrombin and characterizes a new recognition mechanism on EC for this molecule.     

Long-term prognostic significance of homocysteine in middle-aged and elderly

Objective: We investigated the association among increased levels of plasma homocysteine (Hcy), all-cause mortality, and cardiovascular events. Methods: Hcy was measured in 670 middle-aged and elderly subjects with no previous manifest cardiovascular disease. The follow-up period was 15 years. Results: Subjects with Hcy?≥?10.8?μmol/l (n?=?231) had a significant higher incidence of all-cause mortality (p?<?0.001) and CV events (p?<?0.001) compared with subjects with Hcy?<?10.8?μmol/l (n?=?439). However, there was no association on high levels of Hcy and VTE events or stroke. Conclusion: Increased levels of Hcy are associated with all-cause mortality and CV events.     

Associations of galectin-3 expression and LGALS-3 (rs4652) gene variant with coronary artery disease risk in diabetics

BackgroundGalectin-3 protein encoded by lectin galactoside-binding soluble-3 (LGALS-3) gene is an important genetic factor in type 2 diabetes mellitus (T2DM) and its cardiovascular obstacles in various populations. We aimed to elicit the pro-inflammatory effect of galectin-3 as determined by interleukin-6 (IL-6) serum levels and to explore the relationship between galectin-3 (LGALS-3 rs4652) gene variant and its expression levels with coronary artery disease (CAD) risk among T2DM Egyptian patients.Methods112 lean subjects were compared to 100 T2DM without CAD and 84 T2DM with CAD. A tetra-primer amplification refractory mutation system polymerase chain reaction was used to test LGALS-3 (rs4652), and galectin3 expression was tested with a quantitative real-time polymerase chain reaction. Serum IL-6 was measured using an enzyme-linked immunosorbent assay.ResultsWe found that the prevalence of LGALS-3 (rs4652) AC genotype and galectin-3 gene expression levels in T2DM with CAD were significantly higher than the additional 2 groups and were correlated positively to IL-6 circulating levels. Also, the C allele carriers (AC+CC) had significantly higher relative Galectin-3 expression levels compared to the A allele carriers (AA).ConclusionsWe concluded that galectin-3 expression levels and LGALS-3 (rs4652) AC genotype were coronary artery disease risk factors in people with type two diabetes among an Egyptian sample.     

Pedogenesis, permafrost, and soil moisture as controlling factors for soil nitrogen and carbon contents across the Tibetan Plateau

We investigated the main parameters [e.g. mean annual air temperature , mean annual soil temperature, mean annual precipitation, soil moisture (SM), soil chemistry, and physics] influencing soil organic carbon (C_org), soil total nitrogen (N_t) as well as plant available nitrogen (N_min) at 47 sites along a 1200 km transect across the high‐altitude and low‐latitude permafrost region of the central‐eastern Tibetan Plateau. This large‐scale survey allows testing the hypothesis that beside commonly used ecological variables, diversity of pedogenesis is another major component for assessing carbon (C) and nitrogen (N) cycling. The aim of the presented research was to evaluate consequences of permafrost degradation for C and N stocks and hence nutrient supply for plants, as the transect covers all types of permafrost including heavily degraded areas and regions without permafrost. Our results show that SM is the dominant parameter explaining 64% of C_org and 60% of N variation. The extent of the effect of SM is determined by permafrost, current aeolian sedimentation occurring mostly on degraded sites, and pedogenesis. Thus, the explanatory power for C and N concentrations is significantly improved by adding CaCO₃ content (P=0.012 for C_org; P=0.006 for N_t) and soil texture (P=0.077 for C_org; P=0.015 for N_t) to the model. For soil temperature, no correlations were detected indicating that in high‐altitude grassland ecosystems influenced by permafrost, SM overrides soil temperature as the main driving parameter at landscape scale. It was concluded from the current study that degradation of permafrost and corresponding changes in soil hydrology combined with a shift from mature stages of pedogenesis to initial stages, have severe impact on soil C and plant available N. This may alter biodiversity patterns as well as the development and functioning of the ecosystems on the Tibetan Plateau.     

Higher Memory Responses in HIV-Infected and Kidney Transplanted Patients than in Healthy Subjects following Priming with the Pandemic Vaccine

Background

Memory responses require immune competence. We assessed the influence of priming with AS03-adjuvanted pandemic vaccine (Pandemrix®) on memory responses of HIV patients, kidney recipients (SOT) and healthy controls (HC).

Method

Participants (HIV: 197, SOT: 53; HC: 156) were enrolled in a prospective study and 390/406 (96%) completed it. All had been primed in 2009/2010 with 1 (HC) or 2 (patients) doses of Pandemrix®, and were boosted with the 2010/2011 seasonal influenza vaccine. Geometric mean titres and seroprotection rates were measured 12 months after priming and 4 weeks after boosting. Primary and memory responses were directly compared in 191 participants (HCW: 69, HIV: 71, SOT: 51) followed during 2 consecutive seasons.

Results

Most participants (HC: 77.8%, HIV: 77.6%, SOT: 66%) remained seroprotected at 12 months post-priming. Persisting A/09/H1N1 titers were high in HIV (100.2) and HC (120.1), but lower in SOT (61.4) patients. Memory responses reached higher titers in HIV (507.8) than in HC (253.5) and SOT (136.9) patients. Increasing age and lack of HAART reduced persisting and memory responses, mainly influenced by residual antibody titers. Comparing 2009/2010 and 2010/2011 titers in 191 participants followed for 2 seasons indicated lower post-2010/2011 titers in HC (240.2 vs 313.9), but higher titers in HIV (435.7 vs 338.0) and SOT (136 vs 90.3) patients.

Conclusions

Priming with 2 doses of Pandemrix® elicited persistent antibody responses and even stronger memory responses to non-adjuvanted seasonal vaccine in HIV patients than 1 dose in healthy subjects. Adjuvanted influenza vaccines may improve memory responses of immunocompromised patients.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT01022905","term_id":"NCT01022905"}}NCT01022905