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To improve our knowledge of the migration pathway of a highly threatened fish species along the Mekong River, strontium isotope ratios (87Sr/86Sr) and 18 trace element concentrations were measured in the water and in the otoliths of an anadromous catfish, Pangasius krempfi, to infer its natal origin and potential migration pathways. Water was sampled at 18 locations along the mainstream, tributaries and distributaries of the Mekong River. To check for accuracy and precision, measurements of the 87Sr/86Sr ratios and trace element concentrations were then compared in two laboratories that use different analytical methods. Differences in trace element concentrations between locations were not significant and could not, therefore, be used to discriminate between migration pathways. However, the Mekong mainstream, tributaries and distributaries could all be discriminated using Sr isotopes. The 87Sr/86Sr profiles recorded in P. krempfi otoliths showed that there were three contingents with obligate freshwater hatching and variable spawning sites along the Mekong mainstream, from Phnom Penh (Cambodia) to Nong Khai (Thailand) or further. After hatching, the fish migrated more or less rapidly to the Mekong Delta and then settled for most of their lifetime in brackish water. Spawning habitats and migration routes may be threatened by habitat shifts and the increasing number of hydropower dams along the river, especially the contingents born above Khone Falls (Laos). The conservation of P. krempfi, as well as other migratory fish in the Mekong River, requires agreements, common actions and management by all countries along the Mekong River. This study highlighted the importance of using both Sr/Ca and 87Sr/86Sr ratios to understand life history of anadromous fishes as the 87Sr/86Sr ratio in the water was shown to be less effective than the Sr/Ca ratio in identifying movements between different saline areas., Competing Interests: The authors have declared that no competing interests exist.

Background: Acute mountain sickness (AMS) occurs in non-acclimatized people after an acute ascent to an altitude of 2,500 m or higher. The aim of this study was to examine the incidence and severity of AMS and associated symptoms in children., Methods: The prospective observational study included 197 healthy, non-acclimatized 11 and 12-year-old children trekking the round-trip from the trailhead to the summit of Xue Mountain, Taiwan (2,179 m to 3,886 m) over 3 days. AMS was evaluated at Qika Hut (2,460 m) on Day 1, at Sanliujiu Hut on Day 2 (3,100 m), and at the same altitude (3,100 m) after reaching the summit on Day 3. We used the Lake Louise Score (LLS) to diagnose AMS and record daily AMS-associated symptoms. We gave acetazolamide to children with mild to moderate AMS. Dexamethasone was reserved for individuals suffering from severe AMS. Acetaminophen was administrated to children with headache, and metoclopramide for those with nausea or vomiting., Results: There were 197 subjects eligible for analysis. The overall incidence of AMS was 40.6%, which was higher in males and in subjects with a higher body mass index (BMI) (p < 0.05). The prevalence of AMS on Day 1 was 5.6%, which was significantly lower than that on Day 2 (29.4%) and Day 3 (23.4%). The mean LLS of all subjects was 1.77 ± 2.08. The overall incidence of severe AMS (LLS ≥ 5) was 12.5%. The mean LLS of the AMS group (3.02 ± 2.46) was significantly higher than that of the non-AMS group (0.92 ± 1.16, p < 0.001). Among the AMS group, the mean LLS was 1.00 ± 1.55 on Day 1, 4.09 ± 1.97 on Day 2, and 3.98 ± 2.42 on Day 3. The most common symptom was sleep disturbance followed by dizziness, and headache. The prevalence of headache was 46.2% on Day 2 at 3,100 m, and 31.3% on Day 3 at the same altitude after climbing the summit (3,886 m). Males experienced significantly more headache and fatigue than females (p < 0.05). The LLS and prevalence of all AMS symptoms were significantly higher in the AMS than the non-AMS group (p < 0.05)., Conclusions: The AMS incidence among children trekking to Xue Mountain was 40.6%. AMS is common and mostly manifests as mild symptoms. Gender (male) and a higher BMI could be considered two independent risk factors of higher AMS incidence. Sleep disturbance is the most common symptom, and the lower prevalence of headache on Day 3 may be due to the effects of medication and/or acclimatization.

Koelreuteria henryi Dummer, an endemic plant of Taiwan, has been used as a folk medicine for the treatment of hepatitis, enteritis, cough, pharyngitis, allergy, hypertension, hyperlipidemia, and cancer. Austrobailignan-1, a natural lignan derivative isolated from Koelreuteria henryi Dummer, has anti-oxidative and anti-cancer properties. However, the effects of austrobailignan-1 on human cancer cells have not been studied yet. Here, we showed that austrobailignan-1 inhibited cell growth of human non-small cell lung cancer A549 and H1299 cell lines in both dose- and time-dependent manners, the IC50 value (48 h) of austrobailignan-1 were 41 and 22 nM, respectively. Data from flow cytometric analysis indicated that treatment with austrobailignan-1 for 24 h retarded the cell cycle at the G2/M phase. The molecular event of austrobailignan-1-mediated G2/M phase arrest was associated with the increase of p21Waf1/Cip1 and p27Kip1 expression, and decrease of Cdc25C expression. Moreover, treatment with 100 nM austrobailignan-1 for 48 h resulted in a pronounced release of cytochrome c followed by the activation of caspase-2, -3, and -9, and consequently induced apoptosis. These events were accompanied by the increase of PUMA and Bax, and the decrease of Mcl-1 and Bcl-2. Furthermore, our study also showed that austrobailignan-1 was a topoisomerase 1 inhibitor, as evidenced by a relaxation assay and induction of a DNA damage response signaling pathway, including ATM, and Chk1, Chk2, γH2AX phosphorylated activation. Overall, our results suggest that austrobailignan-1 is a novel DNA damaging agent and displays a topoisomerase I inhibitory activity, causes DNA strand breaks, and consequently induces DNA damage response signaling for cell cycle G2/M arrest and apoptosis in a p53 independent manner.

Overproduction of N-terminal pyroglutamate (pGlu)-modified proteins utilizing Escherichia coli or eukaryotic cells is a challenging work owing to the fact that the recombinant proteins need to be recovered by proteolytic removal of fusion tags to expose the N-terminal glutaminyl or glutamyl residue, which is then converted into pGlu catalyzed by the enzyme glutaminyl cyclase. Herein we describe a new method for production of N-terminal pGlu-containing proteins in vivo via intracellular self-cleavage of fusion tags by tobacco etch virus (TEV) protease and then immediate N-terminal cyclization of passenger target proteins by a bacterial glutaminyl cyclase. To combine with the sticky-end PCR cloning strategy, this design allows the gene of target proteins to be efficiently inserted into the expression vector using two unique cloning sites (i.e., SnaB I and Xho I), and the soluble and N-terminal pGlu-containing proteins are then produced in vivo. Our method has been successfully applied to the production of pGlu-modified enhanced green fluorescence protein and monocyte chemoattractant proteins. This design will facilitate the production of protein drugs and drug target proteins that possess an N-terminal pGlu residue required for their physiological activities.

Poly (ADP-ribose) polymerase-1 (PARP-1) and telomerase, as well as DNA damage response pathways are targets for anticancer drug development, and specific inhibitors are currently under clinical investigation. The purpose of this work is to evaluate anticancer activities of anthraquinone-derived tricyclic and tetracyclic small molecules and their structure-activity relationships with PARP-1 inhibition in non-small cell lung cancer (NSCLC) and NSCLC-overexpressing Oct4 and Nanog clone, which show high-expression of PARP-1 and more resistance to anticancer drug. We applied our library selected compounds to NCI's 60 human cancer cell-lines (NCI-60) in order to generate systematic profiling data. Based on our analysis, it is hypothesized that these drugs might be, directly and indirectly, target components to induce mitochondrial permeability transition and the release of pro-apoptotic factors as potential anti-NSCLC or PARP inhibitor candidates. Altogether, the most active NSC747854 showed its cytotoxicity and dose-dependent PARP inhibitory manner, thus it emerges as a promising structure for anti-cancer therapy with no significant negative influence on normal cells. Our studies present evidence that telomere maintenance should be taken into consideration in efforts not only to overcome drug resistance, but also to optimize the use of telomere-based therapeutics. These findings will be of great value to facilitate structure-based design of selective PARP inhibitors, in general, and telomerase inhibitors, in particular. Together, the data presented here expand our insight into the PARP inhibitors and support the resource-demanding lead optimization of structurally related small molecules for human cancer therapy.

Protein-protein interactions are critical determinants in biological systems. Engineered proteins binding to specific areas on protein surfaces could lead to therapeutics or diagnostics for treating diseases in humans. But designing epitope-specific protein-protein interactions with computational atomistic interaction free energy remains a difficult challenge. Here we show that, with the antibody-VEGF (vascular endothelial growth factor) interaction as a model system, the experimentally observed amino acid preferences in the antibody-antigen interface can be rationalized with 3-dimensional distributions of interacting atoms derived from the database of protein structures. Machine learning models established on the rationalization can be generalized to design amino acid preferences in antibody-antigen interfaces, for which the experimental validations are tractable with current high throughput synthetic antibody display technologies. Leave-one-out cross validation on the benchmark system yielded the accuracy, precision, recall (sensitivity) and specificity of the overall binary predictions to be 0.69, 0.45, 0.63, and 0.71 respectively, and the overall Matthews correlation coefficient of the 20 amino acid types in the 24 interface CDR positions was 0.312. The structure-based computational antibody design methodology was further tested with other antibodies binding to VEGF. The results indicate that the methodology could provide alternatives to the current antibody technologies based on animal immune systems in engineering therapeutic and diagnostic antibodies against predetermined antigen epitopes.

Pole-to-pole oscillations of the Min proteins in Escherichia coli are required for the proper placement of the division septum. Direct interaction of MinE with the cell membrane is critical for the dynamic behavior of the Min system. In vitro, this MinE-membrane interaction led to membrane deformation; however, the underlying mechanism remained unclear. Here we report that MinE-induced membrane deformation involves the formation of an amphipathic helix of MinE(2-9), which, together with the adjacent basic residues, function as membrane anchors. Biochemical evidence suggested that the membrane association induces formation of the helix, with the helical face, consisting of A2, L3, and F6, inserted into the membrane. Insertion of this helix into the cell membrane can influence local membrane curvature and lead to drastic changes in membrane topology. Accordingly, MinE showed characteristic features of protein-induced membrane tubulation and lipid clustering in in vitro reconstituted systems. In conclusion, MinE shares common protein signatures with a group of membrane trafficking proteins in eukaryotic cells. These MinE signatures appear to affect membrane curvature.