Network-based screen in iPSC-derived cells reveals therapeutic candidate for heart valve disease.

Mapping the gene-regulatory networks dysregulated in human disease would allow the design of network-correcting therapies that treat the core disease mechanism. However, small molecules are traditionally screened for their effects on one to several outputs at most, biasing discovery and limiting the likelihood of true disease-modifying drug candidates. Here, we developed a machine-learning approach to identify small molecules that broadly correct gene networks dysregulated in a human induced pluripotent stem cell (iPSC) disease model of a common form of heart disease involving the aortic valve (AV). Gene network correction by the most efficacious therapeutic candidate, XCT790, generalized to patient-derived primary AV cells and was sufficient to prevent and treat AV disease in vivo in a mouse model. This strategy, made feasible by human iPSC technology, network analysis, and machine learning, may represent an effective path for drug discovery.

Role of Chemical Reactivity and Transition State Modeling for Virtual Screening.

Every drug discovery research program involves synthesis of a novel and potential drug molecule utilizing atom efficient, economical and environment friendly synthetic strategies. The current work focuses on the role of the reactivity based fingerprints of compounds as filters for virtual screening using a tool ChemScore. A reactant-like (RLS) and a product- like (PLS) score can be predicted for a given compound using the binary fingerprints derived from the numerous known organic reactions which capture the molecule-molecule interactions in the form of addition, substitution, rearrangement, elimination and isomerization reactions. The reaction fingerprints were applied to large databases in biology and chemistry, namely ChEMBL, KEGG, HMDB, DSSTox, and the Drug Bank database. A large network of 1113 synthetic reactions was constructed to visualize and ascertain the reactant product mappings in the chemical reaction space. The cumulative reaction fingerprints were computed for 4000 molecules belonging to 29 therapeutic classes of compounds, and these were found capable of discriminating between the cognition disorder related and anti-allergy compounds with reasonable accuracy of 75% and AUC 0.8. In this study, the transition state based fingerprints were also developed and used effectively for virtual screening in drug related databases. The methodology presented here provides an efficient handle for the rapid scoring of molecular libraries for virtual screening.

A paradigm shift in the delivery of services for diagnosis of inherited retinal disease.

OBJECTIVES: Current technologies for delivering gene testing are labour-intensive and expensive. Over the last 3 years, new high-throughput DNA sequencing techniques (next generation sequencing; NGS), with the capability to analyse multiple genes or entire genomes, have been rapidly adopted into research. This study examines the possibility of incorporating NGS into a clinical UK service context. METHODS: The study applied NGS of 105 genes to 50 patients known to be affected by inherited forms of blindness in the setting of a UK National Health Service-accredited diagnostic molecular genetics laboratory. The study assessed the ability of an NGS protocol to identify likely disease-causing genetic variants when compared with current methodologies available through UK diagnostic laboratories. RESULTS: Conventional testing is only applicable to the minority of patients with inherited retinal disease and identifies mutations in fewer than one in four of those patients tested. By contrast, the NGS assay is directed at all patients with such disorders and identifies disease-causing mutations in 50--55%, which is a dramatic increase. This includes patients with apparently 'sporadic' disease, and those for whom clinical management and prognosis are altered as a consequence of defining their disease at a molecular level. CONCLUSIONS: The new NGS approach delivers a step change in the diagnosis of inherited eye disease, provides precise diagnostic information and extends the possibility of targeted treatments including gene therapy. The approach represents an exemplar that illustrates the opportunity that NGS provides for broadening the availability of genetic testing. The technology will be applied to many conditions that are associated with high levels of genetic heterogeneity.

Anthocyanin content, lipid peroxidation and cyclooxygenase enzyme inhibitory activities of sweet and sour cherries.

Cherries contain bioactive anthocyanins that are reported to possess antioxidant, anti-inflammatory, anticancer, antidiabetic and antiobese properties. The present study revealed that red sweet cherries contained cyanidin-3-O-rutinoside as major anthocyanin (>95%). The sweet cherry cultivar "Kordia" (aka "Attika") showed the highest cyanidin-3-O-rutinoside content, 185 mg/100 g fresh weight. The red sweet cherries "Regina" and "Skeena" were similar to "Kordia", yielding cyanidin-3-O-rutinoside at 159 and 134 mg/100 g fresh weight, respectively. The yields of cyanidin-3-O-glucosylrutinoside and cyanidin-3-O-rutinoside were 57 and 19 mg/100 g fresh weight in "Balaton" and 21 and 6.2 mg/100 g fresh weight in "Montmorency", respectively, in addition to minor quantities of cyanidin-3-O-glucoside. The water extracts of "Kordia", "Regina", "Glacier" and "Skeena" sweet cherries gave 89, 80, 80 and 70% of lipid peroxidation (LPO) inhibition, whereas extracts of "Balaton" and "Montmorency" were in the range of 38 to 58% at 250 microg/mL. Methanol and ethyl acetate extracts of the yellow sweet cherry "Rainier" containing beta-carotene, ursolic, coumaric, ferulic and cafeic acids inhibited LPO by 78 and 79%, respectively, at 250 microg/mL. In the cyclooxygenase (COX) enzyme inhibitory assay, the red sweet cherry water extracts inhibited the enzymes by 80 to 95% at 250 microg/mL. However, the methanol and ethyl acetate extracts of "Rainier" and "Gold" were the most active against COX-1 and -2 enzymes. Water extracts of "Balaton" and "Montmorency" inhibited COX-1 and -2 enzymes by 84, and 91 and 77, and 87%, respectively, at 250 microg/mL.

Pest-managing activities of plant extracts and anthraquinones from Cassia nigricans from Burkina Faso.

Insecticidal activity of eight plants collected from Burkina Faso was studied using mosquito (Ochlerotatus triseriatus), Helicoverpa zea and Heliothis virescens larvae and adult white fly (Bemisia tabaci). The n-hexane, ethyl acetate and methanol extracts of Pseudocedrela kotschyi, Strophantus hispidus, Securidaca longepedunculata, Sapium grahamii, Swartzia madagascariensis, Cassia nigricans, Jatropha curcas and Datura innoxia were used in this study. Extracts were tested at 250 microg/mL concentration. All three extracts of C. nigricans, J. curcas (skin and seeds) and D. innoxia exhibited 100% mortality on fourth instar mosquito (O. triseriatus) larvae. In addition, the n-hexane and ethyl acetate extracts of S. hispidus, S. longepedunculata, S. grahamii showed 100% mortality. The ethyl acetate extract of S. madagascariensis was the most active on adult white fly and exhibited 80% mortality. Extracts of all other plants exhibited 30-50% mortality on B. tabaci. In the antifeedant assays against H. zea and H. virescens, the MeOH extracts of C. nigricans, S. madagascarensis and S. hispidus were more effective against H. zea as indicated by 74% larval weight reduction as compared to the control. Since C. nigricans is commonly used in West Africa to protect grain storage from insects, we have characterized the insecticidal components present in its extract. Bioassay directed isolation of C. nigricans leaf extract yielded anthraquinones emodin, citreorosein, and emodic acid and a flavonoid, luteolin. Emodin, the most abundant and active anthraquinone in C. nigricans showed approximately 85% mortality on mosquito larvae Anopheles gambiaea and adult B. tabaci at 50 and 25 microg/mL, respectively, in 24 h. These results suggest that the extract of C. nigricans has the potential to be used as an organic approach to manage some of the agricultural pests.

Developmental changes in sinus node function in growing children: an updated analysis.

Sinus node dysfunction (SAND) may be congenital or acquired following injury or surgery for congenital heart lesions. Sinus node function is evaluated by electrophysiological (EP) parameters of corrected sinus node recovery time (CSNRT) and sinoatrial conduction time (SACT). The aim of this study was to determine age- and gender-specific values for CSNRT and SACT in pediatric patients without structural congenital heart disease. Data were collected on 152 patients who underwent an EP study for evaluation of supraventricular tachycardia between 1997 and 2002. All patients received midazolam and propofol and/or isoflurane for sedation and anesthesia, which are known to not affect EP parameters. The age of transition at which CSNRT changed significantly was 14 years (241.5 +/- 102.0 msec in patients younger than 14 years old and 285.6 +/- 144.3 msec in those older than 14 years, p < 0.05). The upper limits of normal CSNRT (mean + 2 SD) were significantly higher (445 vs 275 msec) and the upper limits of normal SACT values were lower (120 vs 200 msec) than the currently used norms in the younger age group. CSNRT values and atrial refractory period values were significantly longer in males compared to age-matched females [278.5 +/- 15.3 VS : 236.4 +/- 13.6 msec (p < 0.05) and 269.0 +/- 4.9 VS: 244.7 +/- 6.8 msec (p < 0.005), respectively]. The new age- and gender-specific values of EP parameters, which reflect sinus node function, may enable more precise recognition of SAND.