Pharmacological studies of lumisantonin derivatives, with special reference to anti-inflammatory effect and to histamine-release inhibitory action

A number of derivatives and related compounds of lumisantonin were submitted to evaluatien for the action of histamine-release inhibition and antiinflammatory effect, as they structurally resemble guaiazulene in which these actions had been proved. Nineteen compounds of these suppressed 50 per cent or more of the increase in urinary excretion of histamine due to ovomucoid injection. Five of them markedly inhibited all the edemas in the rat hind paws induced by local inoculation of dextran, hyaluronidase, histamine, and 5-hydroxytryptamine. Among these compounds, #32(methyl pyrophotosantoninate) showed a superior effect of inhibition than guaiazulene on all of these edemas, although the effects of two drugs were comparable in the case of oral administration. The members showing the edema inhibition likewise evidently protected passive cutaneous anaphylaxis in guinea pigs by the intraperitoneal administration; the effect of #32 was more marked than guaiazulene. This effect could be observed when applied to the skin with an ointment containing the compouhd in a concentration of more than 0.03 per cent 24 hours before. In vitro histamine releases from the minced lung tissue of sensitized guinea pig elicited by antigen and sinomenine were both inhibited by these compounds. These findings indicate that the main sites of the histamine-release inhibition and of the anti-inflammatory effect of these compounds are in the local tissue. Compound #32 failed to show any analgesic effect in mice, but possessed a considerable antipyretic action in rats. Some of the compounds in the tests depressed guinea-pig ileal strip while guaiazulene increased peristalsis, but any of these actions was not recognized with #32 even in a high concentration. Most of the members effective in inhibiting edemas as well as histamine release proved to be less toxic than guaiazulene. #32 was well tolerated in the doses of 6g/kg orally and of 4g/kg intraperitoneally by mice. The growth curves for three weeks of rats practically did not deviate from that of the controls by daily administration of 1g/kg of #32 by stomach tube and there were no gross and microscopical abnormalities in the main organs and blood.</p

Multiple binding modes of a moderate ice-binding protein from a polar microalga

Ice-binding proteins (IBPs) produced by cold-tolerant organisms interact with ice and strongly control crystal growth. The molecular basis for the different magnitudes of activity displayed by various IBPs (moderate and hyperactive) has not yet been clarified. Previous studies questioned whether the moderate activity of some IBPs relies on their weaker binding modus to the ice surface, compared to hyperactive IBPs, rather than relying on binding only to selected faces of the ice crystal. We present the structure of one moderate IBP from the sea-ice diatom Fragilariopsis cylindrus (fcIBP) as determined by X-ray crystallography and investigate the protein's binding modes to the growing ice-water interface using molecular dynamics simulations. The structure of fcIBP is the IBP-1 fold, defined by a discontinuous β-solenoid delimitated by three faces (A, B and C-faces) and braced by an α-helix. The fcIBP structure shows capping loops on both N- and C-terminal parts of the solenoid. We show that the protein adsorbs on both the prism and the basal faces of ice crystals, confirming experimental results. The fcIBP binds irreversibly to the prism face using the loop between the B and the C-faces, involving also the B-face in water immobilization despite its irregular structure. The α-helix attaches the protein to the basal face with a partly reversible modus. Our results suggest that fcIBP has a looser attachment to ice and that this weaker binding modus is the basis to explain the moderate activity of fcIBP

Cardiosphere-derived exosomal microRNAs for myocardial repair in pediatric dilated cardiomyopathy

Although cardiosphere-derived cells (CDCs) improve cardiac function and outcomes in patients with single ventricle physiology, little is known about their safety and therapeutic benefit in children with dilated cardiomyopathy (DCM). We aimed to determine the safety and efficacy of CDCs in a porcine model of DCM and translate the preclinical results into this patient population. A swine model of DCM using intracoronary injection of microspheres created cardiac dysfunction. Forty pigs were randomized as preclinical validation of the delivery method and CDC doses, and CDC-secreted exosome (CDCex)–mediated cardiac repair was analyzed. A phase 1 safety cohort enrolled five pediatric patients with DCM and reduced ejection fraction to receive CDC infusion. The primary endpoint was to assess safety, and the secondary outcome measure was change in cardiac function. Improved cardiac function and reduced myocardial fibrosis were noted in animals treated with CDCs compared with placebo. These functional benefits were mediated via CDCex that were highly enriched with proangiogenic and cardioprotective microRNAs (miRNAs), whereas isolated CDCex did not recapitulate these reparative effects. One-year follow-up of safety lead-in stage was completed with favorable profile and preliminary efficacy outcomes. Increased CDCex-derived miR-146a-5p expression was associated with the reduction in myocardial fibrosis via suppression of proinflammatory cytokines and transcripts. Collectively, intracoronary CDC administration is safe and improves cardiac function through CDCex in a porcine model of DCM. The safety lead-in results in patients provide a translational framework for further studies of randomized trials and CDCex-derived miRNAs as potential paracrine mediators underlying this therapeutic strategy

Intracoronary Autologous Cardiac Progenitor Cell Transfer in Patients With Hypoplastic Left Heart Syndrome (TICAP) : A Prospective Phase 1 Controlled Trial

RATIONALE: Hypoplastic left heart syndrome (HLHS) remains a lethal congenital cardiac defect. Recent studies have suggested that intracoronary administration of autologous cardiosphere-derived cells (CDCs) may improve ventricular function. OBJECTIVE: The aim of this study was to test whether intracoronary delivery of CDCs is feasible and safe in patients with hypoplastic left heart syndrome. METHODS AND RESULTS: Between January 5, 2011, and January 16, 2012, 14 patients (1.8±1.5 years) were prospectively assigned to receive intracoronary infusion of autologous CDCs 33.4±8.1 days after staged procedures (n=7), followed by 7 controls with standard palliation alone. The primary end point was to assess the safety, and the secondary end point included the preliminary efficacy to verify the right ventricular ejection fraction improvements between baseline and 3 months. Manufacturing and intracoronary delivery of CDCs were feasible, and no serious adverse events were reported within the 18-month follow-up. Patients treated with CDCs showed right ventricular ejection fraction improvement from baseline to 3-month follow-up (46.9%±4.6% to 52.1%±2.4%; P=0.008). Compared with controls at 18 months, cardiac MRI analysis of CDC-treated patients showed a higher right ventricular ejection fraction (31.5%±6.8% versus 40.4%±7.6%; P=0.049), improved somatic growth (P=0.0005), reduced heart failure status (P=0.003), and lower incidence of coil occlusion for collaterals (P=0.007). CONCLUSIONS: Intracoronary infusion of autologous CDCs seems to be feasible and safe in children with hypoplastic left heart syndrome after staged surgery. Large phase 2 trials are warranted to examine the potential effects of cardiac function improvements and the long-term benefits of clinical outcomes

Isolation of alveolar epithelial type II progenitor cells from adult human lungs

Resident stem/progenitor cells in the lung are important for tissue homeostasis and repair. However, a progenitor population for alveolar type II (ATII) cells in adult human lungs has not been identified. The aim of this study is to isolate progenitor cells from adult human lungs with the ability to differentiate into ATII cells. We isolated colony-forming cells that had the capability for self-renewal and the potential to generate ATII cells in vitro. These undifferentiated progenitor cells expressed surface markers of mesenchymal stem cells (MSCs) and surfactant proteins associated with ATII cells, such as CD90 and pro-surfactant protein-C (pro-SP-C), respectively. Microarray analyses indicated that transcripts associated with lung development were enriched in the pro-SP-C+/CD90+ cells compared with bone marrow-MSCs. Furthermore, pathological evaluation indicated that pro-SP-C and CD90 double-positive cells were present within alveolar walls in normal lungs, and significantly increased in ATII cell hyperplasias contributing to alveolar epithelial repair in damaged lungs. Our findings demonstrated that adult human lungs contain a progenitor population for ATII cells. This study is a first step toward better understanding of stem cell biology in adult human lung alveoli

Functional annotation of human long noncoding RNAs via molecular phenotyping

Long noncoding RNAs (lncRNAs) constitute the majority of transcripts in the mammalian genomes, and yet, their functions remain largely unknown. As part of the FANTOM6 project, we systematically knocked down the expression of 285 lncRNAs in human dermal fibroblasts and quantified cellular growth, morphological changes, and transcriptomic responses using Capped Analysis of Gene Expression (CAGE). Antisense oligonucleotides targeting the same lncRNAs exhibited global concordance, and the molecular phenotype, measured by CAGE, recapitulated the observed cellular phenotypes while providing additional insights on the affected genes and pathways. Here, we disseminate the largest-todate lncRNA knockdown data set with molecular phenotyping (over 1000 CAGE deep-sequencing libraries) for further exploration and highlight functional roles for ZNF213-AS1 and lnc-KHDC3L-2.Peer reviewe

Ice-binding proteins from the fungus Antarctomyces psychrotrophicus possibly originate from two different bacteria through horizontal gene transfer

Various microbes, including fungi and bacteria, that live in cold environments produce ice-binding proteins (IBPs) that protect them from freezing. Ascomycota and Basidiomycota are two major phyla of fungi, and Antarctomyces psychrotrophicus is currently designated as the sole ascomycete that produces IBP (AnpIBP). However, its complete amino acid sequence, ice-binding property, and evolutionary history have not yet been clarified. Here, we determined the peptide sequences of three new AnpIBP isoforms by total cDNA analysis and compared them with those of other microbial IBPs. The AnpIBP isoforms and ascomycete-putative IBPs were found to be phylogenetically close to the bacterial ones but far from the basidiomycete ones, which is supported by the higher sequence identities to bacterial IBPs than basidiomycete IBPs, although ascomycetes are phylogenetically distant from bacteria. In addition, two of the isoforms of AnpIBP share low sequence identity and are not close in the phylogenetic tree. It is hence presumable that these two AnpIBP isoforms were independently acquired from different bacteria through horizontal gene transfer (HGT), which implies that ascomycetes and bacteria frequently exchange their IBP genes. The non-colligative freezing-point depression ability of AnpIBP was not very high, whereas it exhibited significant abilities of ice recrystallization inhibition, ice shaping, and cryo-protection against freeze-thaw cycles even at submicromolar concentrations. These results suggest that HGT is crucial for the cold-adaptive evolution of ascomycetes, and their IBPs offer freeze resistance to organisms to enable them to inhabit the icy environments of Antarctica. Databases Nucleotide sequence data are available in the DDBJ database under the accession numbers , , for AnpIBP1a, AnpIBP1b, AnpIBP2, respectively

Investigation of Active Principles in Anaphylactic and Peptone Shock in Dogs

In dogs immediately after the shock induced by Witte peptone 300mg/kg or by bovine serum anaphylaxis, the presence of adenine nucleotides or 5-hydroxytryptamine in the blood plasma obtained from the femoral and hepatic veins were examined. But our present methods of spectrophotometry of the deproteinized plasma or paper chromtography of the deproteinized, freeze-dried plasma failed to reveal any increase in either substance of these

Anti-inflammatory Effect and Histamine-release Inhibitory Effect of Some Derivatives of Acetanilide and Pyrazolone

Three acetanilide and two pyrazolene derivatives were tested for anti-inflammatory effect on the edema of rat's hind-paws induced by the local injection of dextran, hyaluronidase, formaldehyde, histamine and 5-hydroxyptamine, and for histamine-release inhibitory effect on the rat receiving an intraperitoneal injection of ovomucoid which increases urinary output of histamine as a result of histamine release in the whole body. Phenacetin, acetanilide, aminopyrine, aminopropylone and butazolidine served as controls. GP-I, one of the pyrazolone derivatives (for formula cf. table 1), showed fairly marked supression on all 5 kinds of edema with effects comparable to aminopyrine and butazolidine. Acetanilide derivatives tested, also exerted marked inhibition on the edema other than by hyaluronidase or formaldehyde. All the compounds inhibited the histamine release due to ovomucoid injection, but no graded difference could be seen