Influenza Virus Induces Apoptosis via BAD-Mediated Mitochondrial Dysregulation

Influenza virus infection results in host cell death and major tissue damage. Specific components of the apoptotic pathway, a signaling cascade that ultimately leads to cell death, are implicated in promoting influenza virus replication. BAD is a cell death regulator that constitutes a critical control point in the intrinsic apoptosis pathway, which occurs through the dysregulation of mitochondrial outer membrane permeabilization and the subsequent activation of downstream apoptogenic factors. Here we report a novel proviral role for the proapoptotic protein BAD in influenza virus replication. We show that influenza virus-induced cytopathology and cell death are considerably inhibited in BAD knockdown cells and that both virus replication and viral protein production are dramatically reduced, which suggests that virus-induced apoptosis is BAD dependent. Our data showed that influenza viruses induced phosphorylation of BAD at residues S112 and S136 in a temporal manner. Viral infection also induced BAD cleavage, late in the viral life cycle, to a truncated form that is reportedly a more potent inducer of apoptosis. We further demonstrate that knockdown of BAD resulted in reduced cytochrome c release and suppression of the intrinsic apoptotic pathway during influenza virus replication, as seen by an inhibition of caspases-3, caspase-7, and procyclic acidic repetitive protein (PARP) cleavage. Our data indicate that influenza viruses carefully modulate the activation of the apoptotic pathway that is dependent on the regulatory function of BAD and that failure of apoptosis activation resulted in unproductive viral replication.     

Discovery of 4-alkylamino-7-aryl-3-cyanoquinoline LRRK2 kinase inhibitors

Mutations in leucine-rich repeat kinase 2 (LRRK2) are associated with familial Parkinson’s disease (PD). The kinase activity of this complex protein is increased by pathogenic mutations. Inhibition of LRRK2 kinase activity has therefore emerged as a promising approach for the treatment of PD. Herein we report our findings on a series of 4-alkylamino-7-aryl-3-cyanoquinolines that exhibit kinase inhibitory activity against both wild type and G2019S mutant LRRK2. Activity was determined in both biochemical and cellular assays. Compound 14 was further evaluated in an in vivo pharmacodynamic study and found to significantly inhibit Ser935 phosphorylation after oral dosing.     

Estimation of accuracy of patient-specific musculoskeletal modelling: case study on a post polio residual paralysis subject

For patients with patterns ranging out of anthropometric standard values, patient-specific musculoskeletal modelling becomes crucial for clinical diagnosis and follow-up. However, patient-specific modelling using imaging techniques and motion capture systems is mainly subject to experimental errors. The aim of this study was to quantify these experimental errors when performing a patient-specific musculoskeletal model. CT scan data were used to personalise the geometrical model and its inertial properties for a post polio residual paralysis subject. After having performed a gait-based experimental protocol, kinematics data were measured using a VICON motion capture system with six infrared cameras. The musculoskeletal model was computed using a direct/inverse algorithm (LifeMod software). A first source of errors was identified in the segmentation procedure in relation to the calculation of personalised inertial parameters. The second source of errors was subject related, as it depended on the reproducibility of performing the same type of gait. The impact of kinematics, kinetics and muscle forces resulting from the musculoskeletal modelling was quantified using relative errors and the absolute root mean square error. Concerning the segmentation procedure, we found that the kinematics results were not sensitive to the errors (relative error < 1%). However, a strong influence was noted on the kinetics results (deviation up to 71%). Furthermore, the reproducibility error showed a significant influence (relative mean error varying from 5 to 30%). The present paper demonstrates that in patient-specific musculoskeletal modelling variations due to experimental errors derived from imaging techniques and motion capture need to be both identified and quantified. Therefore, the paper can be used as a guideline.     

Structure of the human telomere in Na+ solution: an antiparallel (2+2) G-quadruplex scaffold reveals additional diversity

Single-stranded DNA overhangs at the ends of human telomeric repeats are capable of adopting four-stranded G-quadruplex structures, which could serve as potential anticancer targets. Out of the five reported intramolecular human telomeric G-quadruplex structures, four were formed in the presence of K⁺ ions and only one in the presence of Na⁺ ions, leading often to a perception that this structural polymorphism occurs exclusively in the presence of K⁺ but not Na⁺. Here we present the structure of a new antiparallel (2+2) G-quadruplex formed by a derivative of a 27-nt human telomeric sequence in Na⁺ solution, which comprises a novel core arrangement distinct from the known topologies. This structure complements the previously elucidated basket-type human telomeric G-quadruplex to serve as reference structures in Na⁺-containing environment. These structures, together with the coexistence of other conformations in Na⁺ solution as observed by nuclear magnetic resonance spectroscopy, establish the polymorphic nature of human telomeric repeats beyond the influence of K⁺ ions.     

A new species of Carpinus (Betulaceae) from the Pliocene of Yunnan Province, China

Macrofossils of Carpinus have been widely reported from the Cenozoic of the Northern Hemisphere. However, the leaf cuticules of the genus have rarely been described. A new species, named Carpinus tengchongensis Dai et B.N. Sun, sp. nov., is identified based on 13 leaf fossils, collected from the late Pliocene Mangbang Formation, Tengchong County of Yunnan Province, China. The important characters of the fossil are its oblong-ovate leaf shape, obliquely cordate base, doubly serrulate margin, straight and moderately thick primary vein, pinnate secondary veins, percurrent tertiary veins, orthogonally reticulate areoles, absence of veinlets, anomocytic stomata with double-layered stomatal rim, well-formed T-pieces and trichome bases, which indicates an affinity within the genus Carpinus section Carpinus subsection Monbeigianae, especially with C. tsaiana. Carpinus fossils were present from the Eocene to Pliocene with disjunctive distribution in the North Temperate Zone, which broadly reflected the present distribution pattern and probably demonstrates the tolerance of Carpinus to environmental change.     

Novel URAT1 mutations caused acute renal failure after exercise in two Chinese families with renal hypouricemia

Renal hypouricemia (RHUC), as an infrequent hereditary disease, is associated with severe complications such as exercise-induced acute renal failure (EIARF). Loss-of-function mutations in urate transporter gene URAT1 (Type 1) and in glucose transporter gene GLUT9 (Type 2) are major causes of this disorder. In this study, URAT1 and GLUT9 were screened in two uncorrelated families from mainland China and a total of five mutations were identified in exons, including two novel heterozygous URAT1 mutations. In four members of the first family, c.151delG (p.A51fsX64) in exon 1 was detected, which resulted in a frameshift and truncated the original 553-residue-protein to 63 amino acid protein. A missense mutation c.C1546A (p.P516T) in exon 9 in GLUT9 was revealed in the second family, which caused a functional protein substitution at codon 516. These two novel mutations were neither identified in the subsequent scanning of 200 ethnically matched healthy control subjects with normal serum UA level nor in a 1000 genome project database. Thus our report identifies two novel loss-of-function mutations (c.151delG in URAT1 and p.P516T in GLUT9) which cause RHUC and renal dysfunction in two independent RHUC pedigrees.     

Wnt3a regulates proliferation,apoptosis and function of pancreatic NIT‐1 beta cells via activation of IRS2/PI3K signaling

Wnt‐signaling pathway is implicated in pancreatic development and functional regulation of mature beta‐cells. Wnt3a/Wnt pathway activation expands islet cell mass in vitro by increasing proliferation and decreasing apoptosis of beta‐cells, thereby enhancing its function. However, the signaling pathways that mediate these effects remain unknown. By using a clonal beta‐cell line (NIT‐1), we examined the role of IRS2/PI3K in the mediation of Wnt3a‐stimulated beta‐cell growth. Real‐time PCR and Western blot were employed to investigate the activity of Wnt/β‐catenin and IRS2/PI3K signaling. Proliferation of NIT‐1 cells was assessed by BrdU incorporation, and apoptosis was quantitatively determined by TUNEL and flow cytometry (FCM). Dkk1, an inhibitor of Wnt signaling, and wortmannin, an inhibitor of PI3K, were also used. Results showed that Wnt3a rapidly activated Wnt/β‐catenin signaling, promoted IRS2 expression and Akt phosphorylation in NIT‐1 cells. These effects were completely abrogated by Dkk1 or partially eliminated by wortmannin. Wnt3a also promoted NIT‐1 cell proliferation, inhibited cytokine‐induced beta‐cell apoptosis, and increased insulin secretion. Both of these effects were also eliminated by Dkk1 or wortmannin. Our results demonstrated that Wnt3a regulates proliferation, apoptosis and enhances function of pancreatic NIT‐1 beta cells via activation of Wnt/β‐catenin signaling, involving crosstalk with IRS2/PI3K signaling, with the effect of Wnt signaling on beta‐cells also being IRS2/PI3K/AKT dependent. J. Cell. Biochem. 114: 1488–1497, 2013. © 2013 Wiley Periodicals, Inc.     

Annotating Diseases Using Human Phenotype Ontology Improves Prediction of Disease-Associated Long Non-coding RNAs

Recently, many long non-coding RNAs (lncRNAs) have been identified and their biological function has been characterized; however, our understanding of their underlying molecular mechanisms related to disease is still limited. To overcome the limitation in experimentally identifying disease–lncRNA associations, computational methods have been proposed as a powerful tool to predict such associations. These methods are usually based on the similarities between diseases or lncRNAs since it was reported that similar diseases are associated with functionally similar lncRNAs. Therefore, prediction performance is highly dependent on how well the similarities can be captured. Previous studies have calculated the similarity between two diseases by mapping exactly each disease to a single Disease Ontology (DO) term, and then use a semantic similarity measure to calculate the similarity between them. However, the problem of this approach is that a disease can be described by more than one DO terms. Until now, there is no annotation database of DO terms for diseases except for genes. In contrast, Human Phenotype Ontology (HPO) is designed to fully annotate human disease phenotypes. Therefore, in this study, we constructed disease similarity networks/matrices using HPO instead of DO. Then, we used these networks/matrices as inputs of two representative machine learning-based and network-based ranking algorithms, that is, regularized least square and heterogeneous graph-based inference, respectively. The results showed that the prediction performance of the two algorithms on HPO-based is better than that on DO-based networks/matrices. In addition, our method can predict 11 novel cancer-associated lncRNAs, which are supported by literature evidence.     

A Malian native plant growth promoting Actinomycetes based biofertilizer improves maize growth and yield

This study was carried out to evaluate the effect of Actinomycetes with Plant growoth promoting activity on the growth and yield of maize. This allowed the selection of three Actinomycetes: Actinomycetes sp. H7, O19 and AHB12 for their ability to solubilize phosphates, fix atmospheric nitrogen, and produce antimicrobial substances, enzymes, phytohormones and for their high vigour index. Five seeds, previously sanitized, inoculated and coated, were sown per pot. The best isolates selected in greenhouses were tested in station experiments. In pot, the inoculated seeds, with Actinomycete sp. H7 gave the best growth in plant size, 19.3% more compared to the uninoculated control (P?≤?0.05). In station experiments, it was found that Actinomycete sp. H7 significantly increased the fresh and dry biomass of the aerial part with 919.7 g and 405.6 g, respectively, against 636.70 g and 297.36 g respectively for the control. The best yield of seeds was obtained with the combination O19-AHB12 with a yield of 311.5 g for 1000 seeds compared to 178.28 g for the uninoculated control. All corn seeds inoculated showed better growth than controls. These results confirm the value of the PGPRs and above all open a way for the formulation and the use of biofertilizers based on PGPRs in Mali.