Akt promotes post-irradiation survival of human tumor cells through initiation, progression, and termination of DNA-PKcs-dependent DNA double-strand break repair

Akt phosphorylation has previously been described to be involved in mediating DNA damage repair through the nonhomologous end-joining (NHEJ) repair pathway. Yet the mechanism how Akt stimulates DNA-protein kinase catalytic subunit (DNA-PKcs)-dependent DNA double-strand break (DNA-DSB) repair has not been described so far. In the present study, we investigated the mechanism by which Akt can interact with DNA-PKcs and promote its function during the NHEJ repair process. The results obtained indicate a prominent role of Akt, especially Akt1 in the regulation of NHEJ mechanism for DNA-DSB repair. As shown by pull-down assay of DNA-PKcs, Akt1 through its C-terminal domain interacts with DNA-PKcs. After exposure of cells to ionizing radiation (IR), Akt1 and DNA-PKcs form a functional complex in a first initiating step of DNA-DSB repair. Thereafter, Akt plays a pivotal role in the recruitment of AKT1/DNA-PKcs complex to DNA duplex ends marked by Ku dimers. Moreover, in the formed complex, Akt1 promotes DNA-PKcs kinase activity, which is the necessary step for progression of DNA-DSB repair. Akt1-dependent DNA-PKcs kinase activity stimulates autophosphorylation of DNA-PKcs at S2056 that is needed for efficient DNA-DSB repair and the release of DNA-PKcs from the damage site. Thus, targeting of Akt results in radiosensitization of DNA-PKcs and Ku80 expressing, but not of cells deficient for, either of these proteins. The data showed indicate for the first time that Akt through an immediate complex formation with DNA-PKcs can stimulate the accumulation of DNA-PKcs at DNA-DSBs and promote DNA-PKcs activity for efficient NHEJ DNA-DSB repair.     

Conversion of cholesterol to progesterone by turtle corpus luteum

Homogenates of corpora lutea from the snapping turtle, $\text{Chelydra}$$\text{serpentina serpentina}$ were capable of converting small (< 1%) amounts of cholesterol-³H to progesterone-³H. There was about twice as much steroidogenic activity in corpora lutea taken from animals still carrying oviducal eggs as in those from animals that had laid their eggs. However, the latter showed up to an 80% increase in conversion of cholesterol to progesterone when turtle pituitary homogenate was incubated along with the luteal homogenate. Approximately 4 and 9 mg/g of free and esterified sterol, respectively, were found in the turtle corpus luteum. These results suggest that the corpus luteum of the oviparous turtle functions as a steroidogenic organ.     

Current Prevalence Pattern of Hypertension in Nigeria: A Systematic Review

BackgroundThe global burden of hypertension and other non-communicable diseases (NCDs) is rapidly increasing, and the African continent seems to be the most affected region in the world. The prevalence of hypertension in Nigeria forms a substantial portion of the total burden in Africa because of the large population of the country currently estimated to be over 170 million.ObjectiveThe purpose of this systematic review is to summarise up to date data on the prevalence and distribution of hypertension in Nigeria from prevalence studies.MethodsA search of the following databases: PubMed, EMBase and WHO cardiovascular InfoBase from 1968 till date was conducted to identify studies which provide estimates of prevalence of hypertension in Nigeria.ResultsThe search yielded a total of 1748 hits from which 45 relevant studies met the inclusion criteria for the review. The overall crude prevalence of hypertension ranged from 0.1% (95%CI:-0.1 to 0.3) to 17.5% (95% CI: 13.6 to 21.4) in children and 2.1% (95%CI: 1.4 to 2.8) to 47.2% (95%CI: 43.6 to 50.8) in adults depending on the benchmark used for diagnosis of hypertension, the setting in which the study was conducted, sex and ethnic group. The crude prevalence of hypertension ranged from 6.2% (95%CI: 4.0 to 8.4) to 48.9% (95%CI: 42.3 to 55.5) for men and 10% (95%CI: 8.1 to 12) to 47.3% (95%CI: 43 to 51.6%) for women. In most studies, prevalence of hypertension was higher in males than females. In addition, prevalence across urban and rural ranged from 9.5% (95%CI: 13.6 to 21.4) to 51.6% (95%CI: 49.8 to 53.4) and 4.8% (95%CI: 2.9 to 6.7) to 43% (95%CI: 42.1 to 43.9) respectively.ConclusionsThe prevalence of hypertension is high among the Nigerian population. Appropriate interventions need to be developed and implemented to reduce the preventable burden of hypertension especially at Primary Health Care Centres which is the first point of call for over 55% of the Nigerian population.     

Detection of Cryptosporidium parvum Oocysts on Fresh Produce Using DNA Aptamers

There are currently no standard methods for the detection of Cryptosporidium spp., or other protozoan parasites, in foods, and existing methods are often inadequate, with low and variable recovery efficiencies. Food testing is difficult due to the low concentrations of parasites, the difficulty in eluting parasites from some foods, the lack of enrichment methods, and the presence of PCR inhibitors. The main objectives of the present study were to obtain DNA aptamers binding to the oocyst wall of C. parvum, and to use the aptamers to detect the presence of this parasite in foods. DNA aptamers were selected against C. parvum oocysts using SELEX (Systematic Evolution of Ligands by EXponential enrichment). Ten rounds of selection led to the discovery of 14 aptamer clones with high affinities for C. parvum oocysts. For detecting parasite-bound aptamers, a simple electrochemical sensor was employed, which used a gold nanoparticle-modified screen-printed carbon electrode. This aptasensor was fabricated by self-assembling a hybrid of a thiolated ssDNA primer and the anti- C. parvum aptamer. Square wave voltammetry was employed to quantitate C. parvum in the range of 150 to 800 oocysts, with a detection limit of approximately 100 oocysts. The high sensitivity and specificity of the developed aptasensor suggests that this novel method is very promising for the detection and identification of C. parvum oocysts on spiked fresh fruits, as compared to conventional methods such as microscopy and PCR.     

Adenovirus Entry From the Apical Surface of Polarized Epithelia Is Facilitated by the Host Innate Immune Response

Prevention of viral-induced respiratory disease begins with an understanding of the factors that increase or decrease susceptibility to viral infection. The primary receptor for most adenoviruses is the coxsackievirus and adenovirus receptor (CAR), a cell-cell adhesion protein normally localized at the basolateral surface of polarized epithelia and involved in neutrophil transepithelial migration. Recently, an alternate isoform of CAR, CAR^Ex8, has been identified at the apical surface of polarized airway epithelia and is implicated in viral infection from the apical surface. We hypothesized that the endogenous role of CAR^Ex8 may be to facilitate host innate immunity. We show that IL-8, a proinflammatory cytokine and a neutrophil chemoattractant, stimulates the protein expression and apical localization of CAR^Ex8 via activation of AKT/S6K and inhibition of GSK3β. Apical CAR^Ex8 tethers infiltrating neutrophils at the apical surface of a polarized epithelium. Moreover, neutrophils present on the apical-epithelial surface enhance adenovirus entry into the epithelium. These findings suggest that adenovirus evolved to co-opt an innate immune response pathway that stimulates the expression of its primary receptor, apical CAR^Ex8, to allow the initial infection the intact epithelium. In addition, CAR^Ex8 is a new target for the development of novel therapeutics for both respiratory inflammatory disease and adenoviral infection.     

Elevated Circulating Interleukin 33 Levels in Stable Renal Transplant Recipients at High Risk for Cardiovascular Events

Background

The Major Adverse Cardiovascular Events calculator (CRCRTR-MACE) estimates the burden of cardiovascular risk in renal transplant recipients (RTR). Our recent study of 95 RTR reported the 7-year median risk of cardiovascular events (CVE) to be 9.97%, ranging from 1.93 to 84.27%. Nearly a third (28.4%) of the cohort was above 20% risk for a CVE. Since interleukins (ILs) as part of the inflammatory response may play a role in the pathogenesis of cardiovascular disease (CVD), we extended this study to identify which ILs are associated with high cardiovascular risk in this population.

Methods

Twenty-two ILs were measured by multiplexed fluorescent bead-based immunoassay in 95 RTR and 56 normal controls. Stepwise analysis after multivariate determination of significant demographic and inflammatory variables was performed between the high and low-CVD risk groups (which were arbitrarily set at scores <10% and ≥20%, respectively). Normalized data was presented as mean ± SD and non-normalized data as median (minimum–maximum). Significance was measured at <0.05.

Results

27.5% of the low-risk and 31.3% of the high-risk groups had mean IL levels above the 95 percentile of the normal control levels. In the non-parametric analysis IL-6, 9, 16, 17 and 33 were significantly higher in the high-risk group compared to the control. Univariate analysis (UVA) of the high-risk group identified IL-33 as the only IL that remained significantly higher than the control and low-risk groups (p = 0.000). The percentage of patients with IL-33 levels above the 90 percentile of control value in the low and high-risk groups were 15.6% and 52.0%, respectively (p<0.002). UVA of factors significant to high IL-33 levels included estimated glomerular filtration rate (eGFR), while diabetes mellitus, serum phosphorus, microalbuminuria and age also remained significant in the multivariate analysis.

Conclusion

Circulating IL-33 level is positively associated with high CRCRTR-MACE score. Diminished eGFR, age, diabetes, serum phosphorus and microalbuminurea demonstrate significant relationship with elevated IL-33 levels, supporting the possible pathognomonic role of IL-33 in the cardiovascular burden in RTR.     

Hydrological Response to Land Cover Changes and Human Activities in Arid Regions Using a Geographic Information System and Remote Sensing

The hydrological response to land cover changes induced by human activities in arid regions has attracted increased research interest in recent decades. The study reported herein assessed the spatial and quantitative changes in surface runoff resulting from land cover change in the Al-Baha region of Saudi Arabia between 1990 and 2000 using an ArcGIS-surface runoff model and predicted land cover and surface runoff depth in 2030 using Markov chain analysis. Land cover maps for 1990 and 2000 were derived from satellite images using ArcGIS 10.1. The findings reveal a 26% decrease in forest and shrubland area, 28% increase in irrigated cropland, 1.5% increase in sparsely vegetated land and 0.5% increase in bare soil between 1990 and 2000. Overall, land cover changes resulted in a significant decrease in runoff depth values in most of the region. The decrease in surface runoff depth ranged from 25-106 mm/year in a 7020-km² area, whereas the increase in such depth reached only 10 mm/year in a 243-km² area. A maximum increase of 73 mm/year was seen in a limited area. The surface runoff depth decreased to the greatest extent in the central region of the study area due to the huge transition in land cover classes associated with the construction of 25 rainwater harvesting dams. The land cover prediction revealed a greater than twofold increase in irrigated cropland during the 2000-2030 period, whereas forest and shrubland are anticipated to occupy just 225 km² of land area by 2030, a significant decrease from the 747 km² they occupied in 2000. Overall, changes in land cover are predicted to result in an annual increase in irrigated cropland and dramatic decline in forest area in the study area over the next few decades. The increase in surface runoff depth is likely to have significant implications for irrigation activities.     

Low concentration structural dynamics of lanreotide and somatostatin‐14

Lanreotide, a synthetic cyclic octapeptide, analogue of the peptide hormone somatostatin‐14 (SST‐14), is routinely used as a long‐acting medication in the management of neuroendocrine tumors. Despite its therapeutic importance, low concentration structural data is still lacking for lanreotide. In fact, the major part of the previous structural investigations were focused on the remarkable aggregation properties of this peptide, appearing at high concentrations (>5 mM). Here, we have applied three optical spectroscopic techniques, i.e. fluorescence, circular dichroism and Raman scattering, for analyzing the structural dynamics at the concentrations below 5 mM, where lanreotide exists either in a monomer state or at the first stages of aggregation. The obtained data from lanreotide were discussed through their comparison with those collected from SST‐14, leading us to the following conclusions: (i) The central D‐Trp residue, forming with its adjacent Lys the main receptor interacting part of lanreotide, keeps a constant high rotational freedom whatever the environment (water, water/methanol, methanol). (ii) A solvent‐dependent tight β‐turn, belonging to the type‐II' family, is revealed in lanreotide. (iii) Raman data analyzed by band decomposition in the amide (I and III) regions allowed estimation of different secondary structural elements within the millimolar range. Interestingly, the applied protocol shows a perfect agreement between the structural features provided by the amide I and amide III Raman markers. © 2014 Wiley Periodicals, Inc. Biopolymers 101: 1019–1028, 2014.     

Facies analysis of a large-scale Jurassic shelf-lagoon: the Kamar-e-Mehdi Formation of east-central Iran

The Callovian–Lower Kimmeridgian Kamar-e-Mehdi Formation of the Tabas Block (east-central Iran) is an up to 1,350-m-thick, fine-grained, marly-calcareous unit containing a basal Echellon Limestone Member (up to 180 m thick) and a terminal Nar Limestone Member (up to 100 m thick). The formation was deposited in a relatively deep shelf-lagoon that was part of the large-scale carbonate system of the Esfandiar Subgroup, extending N–S for about 500 km along the strike with a width of up to 100 km. The lagoonal Kamar-e-Mehdi Formation shows sedimentation rates of 150 m/myr, twice as high as those of the shelf-edge carbonate barrier (Esfandiar Platform). The repetitive lithologies and uniform depositional environment suggest equilibrium conditions between sedimentation and subsidence, related to constant slow rotation of the Tabas fault-block around a horizontal axis, the platform sitting on the crest, and the lagoon occupying the dip-slope. Lagoonal sedimentation was dominated by suspended carbonate mud and peloids from the eastern Esfandiar Platform whereas the subordinate siliciclastic material was derived from the west (Yazd Block). The diverse macrobenthos (mainly bivalves) suggests fully marine conditions for the major part of the Kamar-e-Mehdi Formation. However, towards the upper part, biotic impoverishment and the deposition of skeletal-poor, evaporitic sediments indicate increasing restriction. The overlying Magu Gypsum Formation marks the end of an arid basin-fill cycle and possibly forms an effective seal for hydrocarbon reservoirs in that area. The Esfandiar Subgroup was a Neotethys-facing carbonate margin, forming part of a belt of carbonate systems tracking the margins of the Iran Plate during Callovian to Late Jurassic times.