Shikonin‐loaded liposomes as a new drug delivery system: Physicochemical characterization and in vitro cytotoxicity

Alkannin and shikonin are naturally occurring hydroxynaphthoquinones with a well‐established spectrum of wound healing, antimicrobial, anti‐inflammatory, and antioxidant activities. Recently, extensive scientific effort has been focused on their effectiveness on several tumors and mechanism(s) of antitumor activity. Liposomes have been proved as adequate drug carriers offering significant advantages over conventional formulations, such as controlled release and targeted drug delivery, leading to the appearance of several liposomal formulations in the market, some of them concerning anticancer drugs. The aim of the present study was to prepare shikonin‐loaded liposomes for the first time in order to enhance shikonin therapeutic index. An optimized technique based on the thin film hydration method was developed and liposomes characterization was performed in terms of their physicochemical characteristics, drug entrapment efficiency, and release profile. Results indicated the successful incorporation of shikonin into liposomes, using both 1,2‐dipalmitoylphosphatidylcholine and egg phosphatidylcholine lipids. Liposomes presented good physicochemical characteristics, high entrapment efficiency and satisfactory in vitro release profile. In vitro cytotoxicity of liposomes was additionally tested against three human cancer cell lines (breast, glioma, and non‐small cell lung cancer) showing a moderate growth inhibitory activity. Practical applications: Shikonin is a naturally occurring hydroxynaphthoquinone and extensive scientific research (in vitro, in vivo, and clinical trials) has been conducted during the last years, focusing on its effectiveness on several tumors and mechanism(s) of antitumor action. The purpose of this work was to prepare and characterize shikonin‐loaded liposomes as a new drug delivery system for shikonin. Liposomal formulations provide significant advantages over conventional dosage forms, such as controlled release and targeted drug delivery for anticancer agents. Thus, liposomes could reduce shikonin's side effects, enhance selectivity to cancer cells and protect shikonin from internal biotransformations and instability matters (oxidization and polymerization). Furthermore, liposomal delivery helps overcome the low aqueous solubility of shikonin, which is the major barrier to its oral and internal administration, since it cannot be dissolved and further absorbed from the receptor.     

Production and characterization of transparent MgAl2O4 prepared by hot pressing

Hot pressing has been investigated for the production of transparent MgAl₂O₄ aimed at the scaling up of the process. Other assessed techniques (hot isostatic pressing, spark plasma sintering) can hardly be used for the production of flat components with large dimensions and good planarity.Hot pressing of stoichiometric Al₂O₃–MgO powder mixtures has been preferred to the direct pressing of spinel powder for the readily availability of pure powders and to exploit the thermodynamic driving force of the spinel formation. LiF has been used as sintering additive.A thermodynamic investigation of the reactions involving LiF, MgO and Al₂O₃ has helped in the comprehension of the densification mechanisms affecting the transparency of spinel. Transparencies up to 70% in the visible range (highest value 78% at 1100 nm) have been obtained. Suitable soakings have been added for promoting the initial liquid phase sintering and the release of LiF through formation of vapour phases.     

Experimental features affecting the transparency of YAG ceramics

This article focuses on the importance of the ceramic production process for the final transparency and overall optical quality of materials to be used as laser hosts. YAG-based ceramics are prepared starting from commercial powders. The materials are prepared by reactive sintering in a clean atmosphere and under high vacuum. Nd or Yb are selected as active elements as the more appropriate for high energy and high peak power lasers. The powder type and treatment and the solvent removal technique are described in detail as well as the experimental conditions adopted during shaping. The influence of the pre-sintering and sintering cycles on the reaction among the involved oxides and on the microstructure after sintering is shown. The optical characterization is also reported.     

HCV RNA levels in serum, liver, and peripheral blood mononuclear cells of chronic hepatitis C patients and their relationship to liver injury

OBJECTIVE: We sought to evaluate the relationship between HCV RNA levels in serum, liver, and peripheral blood mononuclear cells (PBMC) and the degree of liver injury in chronic hepatitis C (CHC) patients. METHODS: Thirty-six consecutive CHC patients were included in the study. The liver damage was evaluated by the histological activity index (HAI) score. The HCV RNA levels in the three compartments studied were assessed by bDNA assay. Nineteen patients were treated with alpha-interferon 2b (IFN). RESULTS: Serum and liver HCV RNA levels in CHC patients were significantly associated with an increasing HAI score irrespective of the HCV genotypes. Cirrhotic patients showed higher HCV RNA levels than the CHC patients with HAI score 1-4 (p < 0.05), but had lower levels than the group with HAI score > 8 (p < 0.03). Patients with HAI score 1-4 showed the lowest levels of HCV RNA in PBMC. There was a strong relation (r = 0.78; p < 0.001) between serum and liver HCV RNA levels, but not between either serum or liver HCV RNA levels and those of PBMC. Seven patients showed a response to IFN and three of these had a sustained response. Pretreatment levels of HCV RNA in PBMC of the IFN responder patients were lower than those of the nonresponder patients (p < 0.02). CONCLUSIONS: The data indicate a relation between serum or liver HCV RNA levels and the degree of liver injury in CHC patients, and show that serum HCV RNA level mirrors the hepatic viral burden.     

Abnormal expression of a 170-kilodalton P-glycoprotein encoded by MDR1 gene, a metabolically active efflux pump, in CD4+ and CD8+ T cells from patients with human immunodeficiency virus type 1 infection

Peripheral blood CD4+ and CD8+ T cells from 16 patients with HIV-1 infection, 8 each with CD4+ T cell counts of > 200/mm3 (group I) and with CD4+ T cell counts of < 200/mm3 (group II), and 8 age- and sex-matched controls, were examined for the expression of P-glycoprotein (P-gp), a 170-kDa phosphoglycoprotein encoded by the MDR1 gene, using dual-color flow cytometric analysis. The function of P-glycoprotein was assessed by the accumulation of rhodamine-123 (Rh123) dye in the presence or absence of cyclosporin A (which inhibits Rh123 efflux). A significantly increased proportion of CD4+ T cells from patients with HIV-1 infection expressed P-glycoprotein as compared to controls, resulting in a significantly increased ratio of the proportions of CD4+P-gp+/CD8+P-gp+ cells. The ratio of CD4+P-gp+/CD8+P-gp+ in group II patients was significantly higher (p = 0.02) than in group I patients, suggesting a progressive increase in P-gp expression with the advancement of HIV-1 infection. The proportions of CD4+P-gp+ and CD8+P-gp+ T cells did not differ significantly between those who received AZT and those who were not treated with AZT. Contrary to expectation, both CD4+ and CD8+ T cells from patients accumulated significantly more Rh123 as compared to controls. Furthermore, cyclosporin A failed to increase intracellular accumulation of Rh123 in CD4+ and CD8+ T cells from patients. These data suggest a functionally defective P-gp expression in HIV-1 infection that appears to increase with the progression of HIV-1 infection. A study of a large number of patients with HIV-1 infection is needed to determine the effects of opportunistic infection and antiretroviral therapy on the expression of P-gp and to determine whether the expression of P-gp could serve as another surrogate marker for the progression of HIV-1 infection.     

simpA: An agent-oriented approach for programming concurrent applications on top of Java

More and more aspects of concurrency and concurrent programming are becoming part of mainstream programming and software engineering, due to several factors such as the widespread availability of multi-core/parallel architectures and Internet-based systems. This leads to the extension of mainstream object-oriented programming languages and platforms-Java is a main example-with libraries providing fine-grained mechanisms and idioms to support concurrent programming, in particular for building efficient programs. Besides this fine-grained support, a main research goal in this context is to devise higher-level, coarse-grained abstractions that would help building concurrent programs, as pure object-oriented abstractions help building large component-based programs. To this end, in this paper we present simpA, a Java-based framework that provides programmers with agent-oriented abstractions on top of the basic OO layer, as a means to organize and structure concurrent applications. We first describe the application programming interface (API) and annotation framework provided to Java programmers for building simpA applications, and then we discuss the main features of the approach from a software engineering point of view, by showing some programming examples. Finally, we define an operational semantics formalizing the main aspects of this programming model.     

Uptake of Escherichia coli, Vibrio cholerae non-O1 and Enterococcus durans by, and depuration of mussels (Mytilus galloprovincialis)

The uptakes of Escherichia coli, Vibrio cholerae non-O1 and Enterococcus durans by mussels (Mytilus galloprovincialis) and the times for depuration were investigated in order to determine the most useful indicator of vibrio contamination. The mussels were maintained in tanks of static seawater contaminated with bacteria at 5 log10 CFU/ml for bioaccumulation. Depuration was carried out by circulating fresh seawater through the tanks. Each organism was presented alone and with others to mussels, at temperatures of 14 and 21 degrees C. In water contaminated with either single or mixed organisms, the bacteria accumulated rapidly in the mussels reaching high concentrations after 1 h. With both single and mixed organisms, the maximum numbers of E. coli in mussels were 6.6 log10 CFU/g at 14 degrees C and 5.4 log10 CFU/g at 21 degrees C. Both V. cholerae non-O1 and E. durans alone or with other organisms reached a number ranging from 6.5 to 7 log10 CFU/g at both temperatures. During depuration the numbers of all the organisms slowly decreased, with E. coli alone, numbers ranged from 2.8 to 2 log10 CFU/g after 72 h at both 14 and 21 degrees C, and the organisms were undetectable after 144 h. With mixed organisms at 14 degrees C E. coli became undetectable after 168 h but at 21 degrees C no E. coli were recovered after 72 h. At 14 degrees C V. cholerae non-O1 alone also was undetectable after 168 h, but at 21 degrees C and with mixed organisms at both temperatures. V. cholerae was recovered after 168 h at numbers about 1 log10 CFU/g. After 168 h numbers of E. durans alone ranged from 2.6 log10 CFU/g at 14 degrees C to 1.5 log10 CFU/g at 21 degrees C, and with mixed organisms the numbers ranged from 2.3 to 2.0 log10 CFU/g at both temperatures. Of the three bacteria of faecal origin, E. durans is quickly acquired by mussels and released more slowly than the others, while E. coli quickly becomes undetectable. The results suggest that, for this kind of seafood, enterococci may be a more appropriate indicator than E. coli of risks to consumers from vibrios.     

Role of powder properties and shaping techniques on the formation of pore-free YAG materials

A key feature of transparent ceramics is the absence of residual porosity because boundary between pores and ceramic grains is the origin for light scattering. Powders characterized by a grain size in the nanometric range are generally adopted for obtaining transparent ceramics because of their superior reactivity, but the formation of undesired secondary phase related to the presence of impurities, is observed. The present study shows the results obtained with alternative, highly pure, micrometric powders with two shaping techniques, cold isostatic pressing (CIP) and slip casting (SC). The powder treatment and shaping process are easier when coarser powders are adopted. The influence of the powder properties and of the dispersant system on the particle packing, on the density and on the porosity are studied in relation to the two shaping techniques. The role of the aforementioned features on the final microstructure and on the optical properties are also discussed.     

Successful treatment with ampicillin and fluoroquinolones of human endocarditis due to high-level gentamicin-resistant enterococci

Two cases of endocarditis, one caused by high-level gentamicin-resistant Enterococcus durans and the other by high-level gentamicin- and glycopeptide-resistant Enterococcus faecalis. successfully treated with a combination of ampicillin and a fluoroquinolone are reported. Both strains were susceptible to ampicillin. Enterococcus faecalis was susceptible to ciprofloxacin and to ofloxacin, but Enterococcus durans was moderately resistant to these agents. Microbiological and clinical cure was obtained with a 6-week course of ampicillin plus ciprofloxacin in one case and with ofloxacin in the second case due to intolerance to ciprofloxacin. The efficacy of the treatment was predicted in vitro by time-kill studies and by adequate serum bactericidal titres.