Dihydrotanshinone,a Natural Diterpenoid,Preserves Blood-Retinal Barrier Integrity via P2X7 Receptor

Activation of P2X7 signaling, due to high glucose levels, leads to blood retinal barrier (BRB) breakdown, which is a hallmark of diabetic retinopathy (DR). Furthermore, several studies report that high glucose (HG) conditions and the related activation of the P2X7 receptor (P2X7R) lead to the over-expression of pro-inflammatory markers. In order to identify novel P2X7R antagonists, we carried out virtual screening on a focused compound dataset, including indole derivatives and natural compounds such as caffeic acid phenethyl ester derivatives, flavonoids, and diterpenoids. Molecular Mechanics/Generalized Born Surface Area (MM/GBSA) rescoring and structural fingerprint clustering of docking poses from virtual screening highlighted that the diterpenoid dihydrotanshinone (DHTS) clustered with the well-known P2X7R antagonist JNJ47965567. A human-based in vitro BRB model made of retinal pericytes, astrocytes, and endothelial cells was used to assess the potential protective effect of DHTS against HG and 2′(3′)-O-(4-Benzoylbenzoyl)adenosine-5′-triphosphate (BzATP), a P2X7R agonist, insult. We found that HG/BzATP exposure generated BRB breakdown by enhancing barrier permeability (trans-endothelial electrical resistance (TEER)) and reducing the levels of ZO-1 and VE-cadherin junction proteins as well as of the Cx-43 mRNA expression levels. Furthermore, HG levels and P2X7R agonist treatment led to increased expression of pro-inflammatory mediators (TLR-4, IL-1β, IL-6, TNF-α, and IL-8) and other molecular markers (P2X7R, VEGF-A, and ICAM-1), along with enhanced production of reactive oxygen species. Treatment with DHTS preserved the BRB integrity from HG/BzATP damage. The protective effects of DHTS were also compared to the validated P2X7R antagonist, JNJ47965567. In conclusion, we provided new findings pointing out the therapeutic potential of DHTS, which is an inhibitor of P2X7R, in terms of preventing and/or counteracting the BRB dysfunctions elicited by HG conditions.     

Endoplasmic reticulum Ca(2+)-ATPase in microsomal vesicles isolated from bovine retinae

Active Ca2+ transport was measured in microsomal vesicles prepared from bovine retinae and was compared with that in disk membranes of the photoreceptor cells of the same retina. The 45Ca uptake was dependent on the presence of Mg(2+)-ATP and was inhibited by vanadate or when GTP substituted for ATP. The dependence of calcium uptake on the external free Ca2+ concentration gave a KM = 13 microM or a KM = 0.1 microM for disks and microsomal vesicles, respectively. A phosphorylated intermediate (E-P) of Ca(2+)-ATPase of about 100 kDa was isolated in microsomal vesicles. The E-P formation was strongly inhibited by thapsigargin and partially by 2,5-di-(-butyl)benzohydroquinone. Digestion of disks or microsomes with calpain had no effect on the phosphorylated intermediate, while digestion with trypsin produced two fragments of approximately 55 kDa and 35 kDa. These results suggest that bovine retinal microsomes contain a calcium pump belonging to the SERCA family.     

Neuroprotective Effects of Testosterone in the Hypothalamus of an Animal Model of Metabolic Syndrome

Metabolic syndrome (MetS) is known to be associated to inflammation and alteration in the hypothalamus, a brain region implicated in the control of several physiological functions, including energy homeostasis and reproduction. Previous studies demonstrated the beneficial effects of testosterone treatment (TTh) in counteracting some MetS symptoms in both animal models and clinical studies. This study investigated the effect of TTh (30 mg/kg/week for 12 weeks) on the hypothalamus in a high-fat diet (HFD)-induced animal model of MetS, utilizing quantitative RT-PCR and immunohistochemical analyses. The animal model recapitulates the human MetS features, including low testosterone/gonadotropin plasma levels. TTh significantly improved MetS-induced hypertension, visceral adipose tissue accumulation, and glucose homeostasis derangements. Within hypothalamus, TTh significantly counteracted HFD-induced inflammation, as detected in terms of expression of inflammatory markers and microglial activation. Moreover, TTh remarkably reverted the HFD-associated alterations in the expression of important regulators of energy status and reproduction, such as the melanocortin and the GnRH-controlling network. Our results suggest that TTh may exert neuroprotective effects on the HFD-related hypothalamic alterations, with positive outcomes on the circuits implicated in the control of energy metabolism and reproductive tasks, thus supporting a possible role of TTh in the clinical management of MetS.     

The AFLATOX® Project: Approaching the Development of New Generation,Natural-Based Compounds for the Containment of the Mycotoxigenic Phytopathogen Aspergillus flavus and Aflatoxin Contamination

The control of the fungal contamination on crops is considered a priority by the sanitary authorities of an increasing number of countries, and this is also due to the fact that the geographic areas interested in mycotoxin outbreaks are widening. Among the different pre- and post-harvest strategies that may be applied to prevent fungal and/or aflatoxin contamination, fungicides still play a prominent role; however, despite of countless efforts, to date the problem of food and feed contamination remains unsolved, since the essential factors that affect aflatoxins production are various and hardly to handle as a whole. In this scenario, the exploitation of bioactive natural sources to obtain new agents presenting novel mechanisms of action may represent a successful strategy to minimize, at the same time, aflatoxin contamination and the use of toxic pesticides. The Aflatox^® Project was aimed at the development of new-generation inhibitors of aflatoxigenic Aspergillus spp. proliferation and toxin production, through the modification of naturally occurring molecules: a panel of 177 compounds, belonging to the thiosemicarbazones class, have been synthesized and screened for their antifungal and anti-aflatoxigenic potential. The most effective compounds, selected as the best candidates as aflatoxin containment agents, were also evaluated in terms of cytotoxicity, genotoxicity and epi-genotoxicity to exclude potential harmful effect on the human health, the plants on which fungi grow and the whole ecosystem.     

Engineering a pharmacologically superior form of leptin for the treatment of obesity

Leptin plays a central role in the homeostasis of body weight through its regulatory effects on appetite and energy expenditure, yet in trials as a therapeutic agent for the treatment of obesity in humans it has been disappointing. The poor clinical efficacy of leptin results from its short circulating half-life, low potency and poor solubility, necessitating large and frequent doses to obtain even modest clinical benefit. Engineered Fc-leptin immunofusins, consisting of the Fc fragment of an immunoglobulin gamma chain followed by leptin, exhibit improved pharmacological properties with very consistent and potent biological activities. Furthermore, in extending the circulating half-life of the protein in vivo from a few minutes for leptin to many hours for Fc-leptin, these proteins have the potential to reduce drastically the dosage and frequency of administration required to obtain clinical benefit. The results of this study show that the engineered leptin immunofusins described here have significantly enhanced pharmacological properties in comparison with the recombinant leptin that was used in clinical trials. As such, they could represent an important step towards a therapeutically superior form of leptin if the disappointing performance of leptin in early clinical trials was due to its poor pharmacological properties rather than any conceptual weakness in the strategy of using leptin for the treatment of obesity and its related disorders.     

Assessment of weld line performance of PP/Talc moldings produced in hot runner injection molds

Weld lines are weak regions in thermoplastic injection moldings caused by low molecular entanglement and unfavorable orientation. Their occurrence may lead to a significantly reduced mechanical performance of the products. Therefore, when weld lines are likely to occur in molded products, they must be taken into account during the mechanical and technological design processes. The weld lines become more critical when particulate fillers are compounded with the polymer. The performance of weld lines in talc‐filled polypropylene box moldings produced with a double‐gated hot runner mold is assessed in this work. The processing conditions were varied in order to cause morphology and tensile‐impact resistance changes. The weld performance at room temperature was assessed in terms of the energy absorbed in the impact tests. It was found that the performance depends on the injection temperature, the injection rate, and the orientation of the talc particles in the weld‐line plane. J. VINYL ADDIT. TECHNOL., 13:159–165, 2007. © 2007 Society of Plastics Engineers     

Enzymatic hydrolysis of cotton wastes. Effect of γ-ray pretreatments

Textile cotton wastes were irradiated with γ-rays at different doses and solubilized with 10% NaOH solutions. The regenerated products underwent enzymatic hydrolysis and their degradation pattern was compared with that of the cotton celluloses untreated or treated only with γ-rays. The different samples were characterized by measurements of X-ray diffraction, degree of polymerization, moisture regain and water retention. The structural and morphological modifications induced by the pretreatments were correlated with the variations of kinetic parameters, particularly the V_max/K_m ratio, defined as specificity constant. Only the combination of irradiation with dissolution pretreatments gave a considerable increase of hydrolysis reaction effectiveness.     

On the human ability to discriminate audio ambiances from similar locations of an urban environment

When developing advanced location-based systems augmented with audio ambiances, it would be cost-effective to use a few representative samples from typical environments for describing a larger number of similar locations. The aim of this experiment was to study the human ability to discriminate audio ambiances recorded in similar locations of the same urban environment. A listening experiment consisting of material from three different environments and nine different locations was carried out with nineteen subjects to study the credibility of audio representations for certain environments which would diminish the need for collecting huge audio databases. The first goal was to study to what degree humans are able to recognize whether the recording has been made in an indicated location or in another similar location, when presented with the name of the place, location on a map, and the associated audio ambiance. The second goal was to study whether the ability to discriminate audio ambiances from different locations is affected by a visual cue, by presenting additional information in form of a photograph of the suggested location. The results indicate that audio ambiances from similar urban areas of the same city differ enough so that it is not acceptable to use a single recording as ambience to represent different yet similar locations. Including an image was found to increase the perceived credibility of all the audio samples in representing a certain location. The results suggest that developers of audio-augmented location-based systems should aim at using audio samples recorded on-site for each location in order to achieve a credible impression.     

Temporal intensity noise characteristics and discrete numeric accuracy of analog liquid-crystal-based spatial light modulators

We investigate the temporal intensity noise characteristics of analog liquid-crystal-based spatial light modulators and how they affect the device's achievable discrete numeric accuracies in an optical computing system. First we present an analytical development that defines the concept of precision in analog computing systems, then we define a noise metric and a precision-optimal quantizer for determining the discrete numeric characteristics of the devices. Second we present an experimental discussion in which a low-noise test facility constructed for this investigation is described, and the noise characteristics of three commercially available liquid-crystal-based modulators are measured and analyzed. The accuracy implications of this measured noise are then discussed within the context of the analytical model for each modulator.