Wound healing and in vitro antiradical activity of five Sedum species grown within two sites of community importance in Emilia-Romagna (Italy)

Sedum genus includes more than 400 different species, many of which having ethnobotany interest. The skin healing is one of the most common therapeutic indication of Sedum spp. In this work, for the first time, we compared five different Sedum species grown in two sites of community importance in Emilia Romagna (Italy): Sedum acre L., Sedum album L., Sedum hispanicum L., Sedum rupestre L. and Sedum sexangulare L., analysing their total phenolic and flavonoid content, their antiradical capacity and the in vitro healing activity on human keratinocytes. Total phenolic content of the five species ranged from 35.41?±?1.18 to 90.22?±?1.03?µg gallic acid equivalent/mg of dry extract, being S. rupestre the richest one. Total flavonoid content ranged from 22.39?±?0.51 to 47.93?±?2.82?µg rutin equivalent/mg of extract and S. album resulted the species with the highest flavonoid content. Antiradical capacity was found to be related to the phenolic content of the extracts. All the extracts were active in wound healing assay and each one showed different kinetic of action and concentration-activity relationship. This study proposes few investigated Sedum species grown in Italy as promising agents for skin healing and suggests further phytochemical and biological investigations.     

Systems biology identifies preserved integrity but impaired metabolism of mitochondria due to a glycolytic defect in Alzheimer's disease neurons

Mitochondrial dysfunction is implicated in most neurodegenerative diseases, including Alzheimer's disease (AD). We here combined experimental and computational approaches to investigate mitochondrial health and bioenergetic function in neurons from a double transgenic animal model of AD (PS2APP/B6.152H). Experiments in primary cortical neurons demonstrated that AD neurons had reduced mitochondrial respiratory capacity. Interestingly, the computational model predicted that this mitochondrial bioenergetic phenotype could not be explained by any defect in the mitochondrial respiratory chain (RC), but could be closely resembled by a simulated impairment in the mitochondrial NADH flux. Further computational analysis predicted that such an impairment would reduce levels of mitochondrial NADH, both in the resting state and following pharmacological manipulation of the RC. To validate these predictions, we utilized fluorescence lifetime imaging microscopy (FLIM) and autofluorescence imaging and confirmed that transgenic AD neurons had reduced mitochondrial NAD(P)H levels at rest, and impaired power of mitochondrial NAD(P)H production. Of note, FLIM measurements also highlighted reduced cytosolic NAD(P)H in these cells, and extracellular acidification experiments showed an impaired glycolytic flux. The impaired glycolytic flux was identified to be responsible for the observed mitochondrial hypometabolism, since bypassing glycolysis with pyruvate restored mitochondrial health. This study highlights the benefits of a systems biology approach when investigating complex, nonintuitive molecular processes such as mitochondrial bioenergetics, and indicates that primary cortical neurons from a transgenic AD model have reduced glycolytic flux, leading to reduced cytosolic and mitochondrial NAD(P)H and reduced mitochondrial respiratory capacity.     

1064‐nm‐resonant gold nanorods for photoacoustic theranostics within permissible exposure limits

Therapeutic and diagnostic methods based on photomechanical effects are attracting much current attention in contexts as oncology, cardiology and vascular surgery, for such applications as photoacoustic imaging or microsurgery. Their underlying mechanism is the generation of ultrasound or cavitation from the interaction of short optical pulses with endogenous dyes or targeted contrast agents. Among the latter, gold nanorods are outstanding candidates, but their use has mainly been reported for photoacoustic imaging and photothermal treatments. Conversely, much less is still known about their value as a precision tool for photomechanical manipulations, such as to impart local damage with high spatial resolution through the expansion and collapse of microbubbles. Here, we address the feasibility of gold nanorods exhibiting a distribution of surface plasmon resonances between about 900 to above 1100 nm as a contrast agent for photoacoustic theranostics. After testing their cytotoxicity and cellular uptake, we discuss their photostability and use to mediate cavitation and the photomechanical destruction of targeted cells. We find that the choice of a plasmonic band peaking around 1064 nm is key to enhance the translational potential of this approach. With respect to the standard alternative of 800 nm, at 1064 nm, relevant regulations on optical exposure are less restrictive and the photonic technology is more mature.      

Rational design,synthesis and characterization of potent,non-peptidic Smac mimics/XIAP inhibitors as proapoptotic agents for cancer therapy

Novel proapoptotic Smac mimics/IAPs inhibitors have been designed, synthesized and characterized. Computational models and structural studies (crystallography, NMR) have elucidated the SAR of this class of inhibitors, and have permitted further optimization of their properties. In vitro characterization (XIAP BIR3 and linker-BIR2–BIR3 binding, cytotox assays, early ADMET profiling) of the compounds has been performed, identifying one lead for further in vitro and in vivo evaluation.     

Dynamic HPLC of stereolabile iron(II) complexes on chiral stationary phases

A series of chiral tris-(1,10)-phenanthroline iron(II) complexes have been resolved by HPLC on chiral stationary phases based on either cellulose tris-(3,5-dimethylphenylcarbamate) or teicoplanin. At sub ambient temperatures, baseline separation of the enantiomers was observed for five different iron(II) complexes featuring substituted phenanthroline ligands. Dynamic HPLC profiles were observed near or above room temperature, indicating on-column Delta/Lambda enantiomerization. Rate constants for the Delta/Lambda interconversion in free solution and during chromatography were obtained by thermal racemization experiments and by computer simulation of the HPLC dynamic plots, respectively.     

HadA is an atypical new multifunctional trimeric coiled-coil adhesin of Haemophilus influenzae biogroup aegyptius, which promotes entry into host cells

The Oca (Oligomeric coiled-coil adhesin) family is a subgroup of the bacterial trimeric autotransporter adhesins, which includes structurally related proteins, such as YadA of Yersinia enterocolitica and NadA of Neisseria meningitidis . In this study, we searched in silico for novel members of this family in bacterial genomes and identified HadA ( Haemophilus adhesin A), a trimeric autotransporter expressed only by Haemophilus influenzae biogroup aegyptius causing Brazilian purpuric fever (BPF), a fulminant septicemic disease of children. By comparative genomics and sequence analysis we predicted that the hadA gene is harboured on a mobile genetic element unique to BPF isolates. Biological analysis of HadA in the native background was limited because this organism is not amenable to genetic manipulation. Alternatively, we demonstrated that expression of HadA confers to a non-invasive Escherichia coli strain the ability to adhere to human cells and to extracellular matrix proteins and to induce in vitro bacterial aggregation and microcolony formation. Intriguingly, HadA is predicted to lack the typical N-terminal head domain of Oca proteins generally associated with cellular receptor binding. We propose here a structural model of the HadA coiled-coil stalk and show that the N-terminal region is still responsible of the binding activity and a KGD motif plays a role. Interestingly, HadA promotes bacterial entry into mammalian cells. Our results show a cytoskeleton re-arrangement and an involvement of clathrin in the HadA-mediated internalization. These data give new insights on the structure-function relationship of oligomeric coiled-coil adhesins and suggest a potential role of this protein in the pathogenesis of BPF.     

Tetrapisispora phaffii killer toxin is a highly specific β-glucanase that disrupts the integrity of the yeast cell wall

Background  

Killer yeasts have been used to combat contaminating wild yeasts in food, to control pathogenic fungi in plants, and in the medical field, to develop novel antimycotics for the treatment of human and animal fungal infections. Among these killer yeasts, Tetrapisispora phaffii (formerly known as Kluyveromyces phaffii) secretes a glycoprotein known as Kpkt that is lethal to spoilage yeasts under winemaking conditions. In the present study, the mode of action of Kpkt, and the specific damage produced by this toxin on sensitive yeasts is investigated.     

Dynamic Interaction of Amphiphysin with N-WASP Regulates Actin Assembly

Amphiphysin 1, an endocytic adaptor concentrated at synapses that couples clathrin-mediated endocytosis to dynamin-dependent fission, was also shown to have a regulatory role in actin dynamics. Here, we report that amphiphysin 1 interacts with N-WASP and stimulates N-WASP- and Arp2/3-dependent actin polymerization. Both the Src homology 3 and the N-BAR domains are required for this stimulation. Acidic liposome-triggered, N-WASP-dependent actin polymerization is strongly impaired in brain cytosol of amphiphysin 1 knock-out mice. FRET-FLIM analysis of Sertoli cells, where endogenously expressed amphiphysin 1 co-localizes with N-WASP in peripheral ruffles, confirmed the association between the two proteins in vivo. This association undergoes regulation and is enhanced by stimulating phosphatidylserine receptors on the cell surface with phosphatidylserine-containing liposomes that trigger ruffle formation. These results indicate that actin regulation is a key function of amphiphysin 1 and that such function cooperates with the endocytic adaptor role and membrane shaping/curvature sensing properties of the protein during the endocytic reaction.     

Vowel Identity between Note Labels Confuses Pitch Identification in Non-Absolute Pitch Possessors

The simplest and likeliest assumption concerning the cognitive bases of absolute pitch (AP) is that at its origin there is a particularly skilled function which matches the height of the perceived pitch to the verbal label of the musical tone. Since there is no difference in sound frequency resolution between AP and non-AP (NAP) musicians, the hypothesis of the present study is that the failure of NAP musicians in pitch identification relies mainly in an inability to retrieve the correct verbal label to be assigned to the perceived musical note. The primary hypothesis is that, when asked to identify tones, NAP musicians confuse the verbal labels to be attached to the stimulus on the basis of their phonetic content. Data from two AP tests are reported, in which subjects had to respond in the presence or in the absence of visually presented verbal note labels (fixed Do solmization). Results show that NAP musicians confuse more frequently notes having a similar vowel in the note label. They tend to confuse e.g. a 261 Hz tone (Do) more often with Sol than, e.g., with La. As a second goal, we wondered whether this effect is lateralized, i.e. whether one hemisphere is more responsible than the other in the confusion of notes with similar labels. This question was addressed by observing pitch identification during dichotic listening. Results showed that there is a right hemispheric disadvantage, in NAP but not AP musicians, in the retrieval of the verbal label to be assigned to the perceived pitch. The present results indicate that absolute pitch has strong verbal bases, at least from a cognitive point of view.