The Essential Autophagy Gene ATG7 Modulates Organ Fibrosis via Regulation of Endothelial-to-Mesenchymal Transition

Pulmonary fibrosis is a progressive disease characterized by fibroblast proliferation and excess deposition of collagen and other extracellular matrix components. Although the origin of fibroblasts is multifactorial, recent data implicate endothelial-to-mesenchymal transition as an important source of fibroblasts. We report herein that loss of the essential autophagy gene ATG7 in endothelial cells (ECs) leads to impaired autophagic flux accompanied by marked changes in EC architecture, loss of endothelial, and gain of mesenchymal markers consistent with endothelial-to-mesenchymal transition. Loss of ATG7 also up-regulates TGFβ signaling and key pro-fibrotic genes in vitro. In vivo, EC-specific ATG7 knock-out mice exhibit a basal reduction in endothelial-specific markers and demonstrate an increased susceptibility to bleomycin-induced pulmonary fibrosis and collagen accumulation. Our findings help define the role of endothelial autophagy as a potential therapeutic target to limit organ fibrosis, a condition for which presently there are no effective available treatments.     

Disruption of NO-cGMP signaling by neonatal hyperoxia impairs relaxation of lung parenchyma

Exposure of immature lungs to hyperoxia for prolonged periods contributes to neonatal lung injury and airway hyperreactivity. We studied the role of disrupted nitric oxide-guanosine 3',5'-cyclic monophosphate (NO-cGMP) signaling in impairing the relaxant responses of lung tissue from hyperoxia-exposed rat pups. Pups were exposed to >/=95% O(2) or room air for 7 days starting from days 1, 5, or 14. The animals were killed, lungs were removed, and 1-mm-thick lung parenchymal strips were prepared. Lung parenchymal strips of room air or hyperoxic pups were preconstricted using bethanechol and then graded electrical field stimulation (EFS) was applied to induce relaxation. EFS-induced relaxation of lung parenchymal strips was greater at 7 and 12 days than at 21 days in room air-exposed rat pups. Hyperoxic exposure significantly reduced relaxation at 7 and 12 days but not 21 days compared with room air exposure. NO synthase blockade with N(omega)-nitro-l-arginine methyl ester diminished relaxant responses in room air but not in hyperoxic pups at 12 days. After incubation with supplemental l-arginine, the relaxation response of hyperoxic strips was restored. cGMP, a key mediator of the NO signaling pathway, also decreased in strips from hyperoxic vs. room air pups and cGMP levels were restored after incubation with supplemental l-arginine. In addition, arginase activity was significantly increased in hyperoxic lung parenchymal strips compared with room air lung parenchymal strips. These data demonstrate disruption of NO-cGMP signaling in neonatal rat pups exposed to hyperoxia and show that bioavailability of the substrate l-arginine is implicated in the predisposition of this model to airway hyperreactivity.     

Maximal eccentric and concentric strength discrepancies between young men and women for dynamic resistance exercise

Although research has demonstrated that isokinetic eccentric (ECC) strength is 20-60% greater than isokinetic concentric (CON) strength, few data exist comparing these strength differences in standard dynamic resistance exercises. The purpose of the study was to determine the difference in maximal dynamic ECC and CON strength for 6 different resistance exercises in young men and women. Ten healthy young men (mean +/- SE, 25.30 +/- 1.34 years), and 10 healthy young women (mean +/- SE, 23.40 +/- 1.37 years) who were regular exercisers with resistance training experience participated in the study. Two sessions were performed to determine CON and ECC 1 repetitions maximum for latissimus pull-down (LTP), leg press (LP), bench press (BP), leg extension (LE), seated military press (MP), and leg curl (LC) exercises. Maximal ECC and maximal CON strength were determined on weight stack machines modified to isolate ECC and CON contractions using steel bars and pulleys such that only 1 type of contraction was performed. Within 2 weeks, participants returned and completed a retest trial in a counterbalanced fashioned. Test-retest reliability was excellent (r = 0.99) for all resistance exercise trials. Men demonstrated 20-60% greater ECC than CON strength (LTP = 32%, LP = 44%, BP = 40%, LE = 35%, MP = 49%, LC = 27%). Women's strength exceeded the proposed parameters for greater ECC strength in 4 exercises, p < 0.05 (LP = 66%, BP = 146%, MP = 161%, LC = 82%). The ECC/CON assessment could help coaches capitalize on muscle strength differences in young men and women during training to aid in program design and injury prevention and to enhance strength development.     

Clinical significance of blood-based miRNAs as diagnostic and prognostic nucleic acid markers in breast cancer: Comparative to conventional tumor markers

microRNAs (miRNAs) are implicated in carcinogenesis and their expression in biological fluids offer great potential as nucleic acid markers for cancer detection and progression. Authors investigated the expression level of miRNAs (miRNA-21, miRNA-126, and miRNA-155) to evaluate their role as diagnostic and prognostic markers for breast cancer compared with other commonly used protein-based markers (CEA and CA15-3). Serum samples from patients with breast cancer (n = 96), patients with benign breast lesion (n = 47), and healthy individuals (n = 39) were enrolled for detection of miRNA expression levels and protein-based tumor markers using fluorescent real-time quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Correlation among investigated markers with clinicopathological factors and clinical outcomes were determined. Expression of miRNA-21 and miRNA-155 revealed significant increases in patients with breast cancer compared with both benign and control groups, the same result was reported for tumor markers; on the other hand, miRNA-126 was significantly decreased in breast cancer group as compared with the other two groups. miRNA frequencies were significantly related to clinical staging and histological grading as compared with tumor markers. Patients with breast cancer with increased miRNA-21 and miRNA-155 and decreased miRNA-126 expressions had significantly worse disease-free survival, while only miRNA-21 and miRNA-126 showed poor OS (P< 0.005). In conclusion, investigated miRNAs were superior over tumor markers for the early stage of breast cancer especially those with high-risk factor and their assessment in blood facilitates their role as a potential prognostic molecular marker.     

Discovery of novel thienoquinoline-2-carboxamide chalcone derivatives as antiproliferative EGFR tyrosine kinase inhibitors

Novel thienoquinoline carboxamide-chalcone derivatives were prepared via the cyclization of acylated chalcones and 2-mercaptoquinoline-3-carbaldehyde in DMF with K₂CO₃. Thienoquinolines 9a–f, h exhibited promising antiproliferative effect against all the tested cell lines and gave a significant activity as EGFR inhibitors, with IC₅₀ values ranging from 0.5 and 3.2?µM, and compounds 9e and 9f being the most active of the series. They also showed better activity than Erlotinib against melanoma cancer cell line A375. Moreover, compound 9f influenced pre G1 apoptosis and cell cycle arrest at G2/M phase. The binding mode of the best EGFR inhibitor 9e in the EGFR active site revealed that the thienoquinoline ring occupied the ATP-binding site while the chalcone moiety is located in the allosteric site and is responsible for the enhanced activity of these compounds.     

Effect of spdC gene expression on virulence and antibiotic resistance in clinical Staphylococcus aureus isolates

International Microbiology - Surface protein display C (SpdC) protein was described as a novel virulence factor of Staphylococcus aureus that affects biofilm formation and pathogenesis and favors...     

Biodiversity,Anti-Trypanosomal Activity Screening,and Metabolomic Profiling of Actinomycetes Isolated from Mediterranean Sponges

Marine sponge–associated actinomycetes are considered as promising sources for the discovery of novel biologically active compounds. In the present study, a total of 64 actinomycetes were isolated from 12 different marine sponge species that had been collected offshore the islands of Milos and Crete, Greece, eastern Mediterranean. The isolates were affiliated to 23 genera representing 8 different suborders based on nearly full length 16S rRNA gene sequencing. Four putatively novel species belonging to genera Geodermatophilus, Microlunatus, Rhodococcus and Actinomycetospora were identified based on a 16S rRNA gene sequence similarity of < 98.5% to currently described strains. Eight actinomycete isolates showed bioactivities against Trypanosma brucei brucei TC221 with half maximal inhibitory concentration (IC₅₀) values <20 μg/mL. Thirty four isolates from the Milos collection and 12 isolates from the Crete collection were subjected to metabolomic analysis using high resolution LC-MS and NMR for dereplication purposes. Two isolates belonging to the genera Streptomyces (SBT348) and Micromonospora (SBT687) were prioritized based on their distinct chemistry profiles as well as their anti-trypanosomal activities. These findings demonstrated the feasibility and efficacy of utilizing metabolomics tools to prioritize chemically unique strains from microorganism collections and further highlight sponges as rich source for novel and bioactive actinomycetes.     

Inflammasome Priming Is Similar for Francisella Species That Differentially Induce Inflammasome Activation

Inflammasome activation is a two-step process where step one, priming, prepares the inflammasome for its subsequent activation, by step two. Classically step one can be induced by LPS priming followed by step two, high dose ATP. Furthermore, when IL-18 processing is used as the inflammasome readout, priming occurs before new protein synthesis. In this context, how intracellular pathogens such as Francisella activate the inflammasome is incompletely understood, particularly regarding the relative importance of priming versus activation steps. To better understand these events we compared Francisella strains that differ in virulence and ability to induce inflammasome activation for their relative effects on step one vs. step two. When using the rapid priming model, i.e., 30 min priming by live or heat killed Francisella strains (step 1), followed by ATP (step 2), we found no difference in IL-18 release, p20 caspase-1 release and ASC oligomerization between Francisella strains (F. novicida, F. holarctica –LVS and F. tularensis Schu S4). This priming is fast, independent of bacteria viability, internalization and phagosome escape, but requires TLR2-mediated ERK phosphorylation. In contrast to their efficient priming capacity, Francisella strains LVS and Schu S4 were impaired in inflammasome triggering compared to F. novicida. Thus, observed differences in inflammasome activation by F. novicida, LVS and Schu S4 depend not on differences in priming but rather on their propensity to trigger the primed inflammasome.     

Isolation of immunologically active uterine luminal proteins associated with follicular and luteal phases of the ovary in buffalo (Bubalus bubalus)

Uterine luminal proteins (ULP) collected from the genital tract of buffalo during the follicular (Group F) and luteal (Group L) phases of the estrous cycle were chromatographed using sephacryl S-200 gel. Five peaks were detected in each group. Different protein concentrations (10 to 200 microg) from Peaks I and V in each group were examined for immunological activity on polymorph nuclear leukocytic cells (PMNL) in vitro. All concentrations except 10 microg of ULP Peak I (< or = 250 kDa) in Group F enhanced phagocytic activity of PMNL. Peak V (56 kDa) in the same group enhanced phagocytic activity of PMNL only at low protein concentrations (10, 20 and 40 microg protein), while at greater concentrations (80, 150 and 200 microg protein) PMNL activity was suppressed. On the other hand, all protein concentrations from Peak 1 (> or = 250 kDa) in Group L suppressed PMNL activity in a dose-dependent manner. Proteins from Peak V (31 kDa) in Group L suppressed PMNL phagocytic activity at all concentrations but not to the same extent as in Peak I. Electrophoretic analysis of Peaks I and V in both groups revealed only 3 detectable protein bands (subunits) in Peak I and 1 detectable subunit in Peak V. Several additional proteins were probably not detected. The molecular weights of the detected subunits in Peaks I and V in Group F were greater than those in Group L as indicated by SDS-PAGE analysis. The results of this study show that ULP collected from buffalo possessed proteins that modulated phagocytic activity of PMNL in vitro. Proteins collected during the follicular phase, especially Peak I, enhanced phagocytic activity of the PMNL, whereas those collected during the luteal phase (Peaks I and V) suppressed activity. Changes in the molecular weights of ULP detected in this experiment may be related to the changes in phagocytic activity of PMNL tested in vitro.