High-throughput synergy screening identifies microbial metabolites as combination agents for the treatment of fungal infections

The high mortality rate of immunocompromised patients with fungal infections and the limited availability of highly efficacious and safe agents demand the development of new antifungal therapeutics. To rapidly discover such agents, we developed a high-throughput synergy screening (HTSS) strategy for novel microbial natural products. Specifically, a microbial natural product library was screened for hits that synergize the effect of a low dosage of ketoconazole (KTC) that alone shows little detectable fungicidal activity. Through screening of approximate to 20,000 microbial extracts, 12 hits were identified with broadspectrum antifungal activity. Seven of them showed little cytotoxicity against human hepatoma cells. Fractionation of the active extracts revealed beauvericin (BEA) as the most potent component, because it dramatically synergized KTC activity against diverse fungal pathogens by a checkerboard assay. Significantly, in our immunocompromised mouse model, combinations of BEA (0.5 mg/kg) and KTC (0.5 mg/kg) prolonged survival of the host infected with Candida parapsilosis and reduced fungal colony counts in animal organs including kidneys, lungs, and brains. Such an effect was not achieved even with the high dose of 50 mg/kg KTC. These data support synergism between BEA and KTC and thereby a prospective strategy for antifungal therapy.

Wintertime meso-scale horizontal distribution of large tintinnids in the southern Yellow Sea

Spatial distribution of some large tintinnid species (nominally > 76 mu m) is investigated on samples vertically towed in the southern Yellow Sea in winters of 2001 to 2004. Nine tintinnid species are recorded: Codonellopsis morchella, Stenosemella pacifica, S. steini, Tintinnopsis schotti, T. radix, T. karajacensis, Eutintinnus tenuis, Parafavella sp., Leprotintinnus neriticus, of which C. morchella and T. radix dominated in the warm tongue-shaped zone of the Yellow Sea Warm Current (YSWC), and S. pacifica is the next in abundance. Our study shows that these tintinnids occur repeatedly in certain special distribution patterns.

Meso-scale spatial distribution of large tintinnids in early summer in southern Yellow Sea

The spatial distribution of some large tintinnid species (nominally>76 mu m) was investigated according to samples collected by vertical towing in cruises to the southern Yellow Sea in summer 2000-2002 and 2004. Eight species were identified: Codonellopsis mobilis, Leprotintinnus netritus, Tintinnopsis karajacensis, T. japonica, T. kiaochowensis, T. butschlii, T. radix, and Parafavella sp. With maximum abundance of 158.2 ind/L in June 2004, C mobilis was the dominant species, lasting from May to July 2004. Tintinnid communities were patchy and distributed mainly in shallow waters along the shore.

Two trichodinid ectoparasites from marine molluscs in the Yellow Sea, off China, with the description of Trichodina caecellae n. sp (Protozoa : Ciliophora : Peritrichia)

The morphology and infraciliature of two ectoparasitic ciliates, Trichodina caecellae n. sp. and T. ruditapicis Xu, Song & Warren, 2000, parasitising the gills of marine molluscs from the Shandong coast of the Yellow Sea, China, were investigated following wet silver nitrate and protargol impregnation. T. caecellae was found on the small marine sand clam Caecella chinensis Deshayes and is distinguished mainly by the acute triangle-like blade, the very delicate central part and the needle-shaped ray. T. ruditapicis was studied based on four populations from three clams: two populations from Ruditapes philippinarum (Adams) and one each from Saxidomus purpuratus (Sowerby) and Solen grandis Dunker. All four populations fell within the range of morphometry and agreed closely in the overall appearance of the adhesive disc. However, variability was found in the denticle structure, especially in populations from different host clams. Our observations suggest that denticle morphology may be more or less variable between and within populations, and that such minor differences should not be overestimated. It should be emphasised that, except for the denticle morphology, the bright granules or circles in the centre of the adhesive disc represent another important feature facilitating the identification of this trichodinid species.

Secure architectures for pairing based public key cryptography

Along with the growing demand for cryptosystems in systems ranging from large servers to mobile devices, suitable cryptogrophic protocols for use under certain constraints are becoming more and more important. Constraints such as calculation time, area, efficiency and security, must be considered by the designer. Elliptic curves, since their introduction to public key cryptography in 1985 have challenged established public key and signature generation schemes such as RSA, offering more security per bit. Amongst Elliptic curve based systems, pairing based cryptographies are thoroughly researched and can be used in many public key protocols such as identity based schemes. For hardware implementions of pairing based protocols, all components which calculate operations over Elliptic curves can be considered. Designers of the pairing algorithms must choose calculation blocks and arrange the basic operations carefully so that the implementation can meet the constraints of time and hardware resource area. This thesis deals with different hardware architectures to accelerate the pairing based cryptosystems in the field of characteristic two. Using different top-level architectures the hardware efficiency of operations that run at different times is first considered in this thesis. Security is another important aspect of pairing based cryptography to be considered in practically Side Channel Analysis (SCA) attacks. The naively implemented hardware accelerators for pairing based cryptographies can be vulnerable when taking the physical analysis attacks into consideration. This thesis considered the weaknesses in pairing based public key cryptography and addresses the particular calculations in the systems that are insecure. In this case, countermeasures should be applied to protect the weak link of the implementation to improve and perfect the pairing based algorithms. Some important rules that the designers must obey to improve the security of the cryptosystems are proposed. According to these rules, three countermeasures that protect the pairing based cryptosystems against SCA attacks are applied. The implementations of the countermeasures are presented and their performances are investigated.

The importance of pH in regulating the function of the Fasciola hepatica cathepsin L1 cysteine protease

The helminth parasite Fasciola hepatica secretes cathepsin L cysteine proteases to invade its host, migrate through tissues and digest haemoglobin, its main source of amino acids. Here we investigated the importance of pH in regulating the activity and functions of the major cathepsin L protease FheCL1. The slightly acidic pH of the parasite gut facilitates the auto-catalytic activation of FheCL1 from its inactive proFheCL1 zymogen; this process was approximately 40-fold faster at pH 4.5 than at pH 7.0. Active mature FheCL1 is very stable at acidic and neutral conditions (the enzyme retained approximately 45% activity when incubated at 37 degrees C and pH 4.5 for 10 days) and displayed a broad pH range for activity peptide substrates and the protein ovalbumin, peaking between pH 5.5 and pH 7.0. This pH profile likely reflects the need for FheCL1 to function both in the parasite gut and in the host tissues. FheCL1, however, could not cleave its natural substrate Hb in the pH range pH 5.5 and pH 7.0; digestion occurred only at pH

Proteomics and phylogenetic analysis of the cathepsin L protease family of the helminth pathogen Fasciola hepatica:Expansion of a repertoire of virulence-associated factors

Cathepsin L proteases secreted by the helminth pathogen Fasciola hepatica have functions in parasite virulence including tissue invasion and suppression of host immune responses. Using proteomics methods alongside phylogenetic studies we characterized the profile of cathepsin L proteases secreted by adult F. hepatica and hence identified those involved in host-pathogen interaction. Phylogenetic analyses showed that the Fasciola cathepsin L gene family expanded by a series of gene duplications followed by divergence that gave rise to three clades associated with mature adult worms (Clades 1, 2, and 5) and two clades specific to infective juvenile stages (Clades 3 and 4). Consistent with these observations our proteomics studies identified representatives from Clades 1, 2, and 5 but not from Clades 3 and 4 in adult F. hepatica secretory products. Clades 1 and 2 account for 67.39 and 27.63% of total secreted cathepsin Ls, respectively, suggesting that their expansion was positively driven and that these proteases are most critical for parasite survival and adaptation. Sequence comparison studies revealed that the expansion of cathepsin Ls by gene duplication was followed by residue changes in the S2 pocket of the active site. Our biochemical studies showed that these changes result in alterations in substrate binding and suggested that the divergence of the cathepsin L family produced a repertoire of enzymes with overlapping and complementary substrate specificities that could cleave host macromolecules more efficiently. Although the cathepsin Ls are produced as zymogens containing a prosegment and mature domain, all secreted enzymes identified by MS were processed to mature active enzymes. The prosegment region was highly conserved between the clades except at the boundary of prosegment and mature enzyme. Despite the lack of conservation at this section, sites for exogenous cleavage by asparaginyl endopeptidases and a Leu-Ser[downward arrow]His motif for autocatalytic cleavage by cathepsin Ls were preserved.

Structural and functional relationships in the virulence-associated cathepsin L proteases of the parasitic liver fluke, Fasciola hepatica

The helminth parasite Fasciola hepatica secretes cysteine proteases to facilitate tissue invasion, migration, and development within the mammalian host. The major proteases cathepsin L1 (FheCL1) and cathepsin L2 (FheCL2) were recombinantly produced and biochemically characterized. By using site-directed mutagenesis, we show that residues at position 67 and 205, which lie within the S2 pocket of the active site, are critical in determining the substrate and inhibitor specificity. FheCL1 exhibits a broader specificity and a higher substrate turnover rate compared with FheCL2. However, FheCL2 can efficiently cleave substrates with a Pro in the P2 position and degrade collagen within the triple helices at physiological pH, an activity that among cysteine proteases has only been reported for human cathepsin K. The 1.4-A three-dimensional structure of the FheCL1 was determined by x-ray crystallography, and the three-dimensional structure of FheCL2 was constructed via homology-based modeling. Analysis and comparison of these structures and our biochemical data with those of human cathepsins L and K provided an interpretation of the substrate-recognition mechanisms of these major parasite proteases. Furthermore, our studies suggest that a configuration involving residue 67 and the "gatekeeper" residues 157 and 158 situated at the entrance of the active site pocket create a topology that endows FheCL2 with its unusual collagenolytic activity. The emergence of a specialized collagenolytic function in Fasciola likely contributes to the success of this tissue-invasive parasite.

A new Nawaz–Enscore–Ham-based heuristic for permutation flow-shop problem with bicriteria of makespan and machine idle-time

A new heuristic based on Nawaz–Enscore–Ham (NEH) algorithm is proposed for solving permutation flowshop scheduling problem in this paper. A new priority rule is proposed by accounting for the average, mean absolute deviation, skewness and kurtosis, in order to fully describe the distribution style of processing times. A new tie-breaking rule is also introduced for achieving effective job insertion for the objective of minimizing both makespan and machine idle-time. Statistical tests illustrate better solution quality of the proposed algorithm, comparing to existing benchmark heuristics.

Feature analysis of spammers in social networks with active honeypots:a case study of Chinese microblogging networks

In this poster we presented our preliminary work on the study of spammer detection and analysis with 50 active honeypot profiles implemented on Weibo.com and QQ.com microblogging networks. We picked out spammers from legitimate users by manually checking every captured user's microblogs content. We built a spammer dataset for each social network community using these spammer accounts and a legitimate user dataset as well. We analyzed several features of the two user classes and made a comparison on these features, which were found to be useful to distinguish spammers from legitimate users. The followings are several initial observations from our analysis on the features of spammers captured on Weibo.com and QQ.com. ¦The following/follower ratio of spammers is usually higher than legitimate users. They tend to follow a large amount of users in order to gain popularity but always have relatively few followers. ¦There exists a big gap between the average numbers of microblogs posted per day from these two classes. On Weibo.com, spammers post quite a lot microblogs every day, which is much more than legitimate users do; while on QQ.com spammers post far less microblogs than legitimate users. This is mainly due to the different strategies taken by spammers on these two platforms. ¦More spammers choose a cautious spam posting pattern. They mix spam microblogs with ordinary ones so that they can avoid the anti-spam mechanisms taken by the service providers. ¦Aggressive spammers are more likely to be detected so they tend to have a shorter life while cautious spammers can live much longer and have a deeper influence on the network. The latter kind of spammers may become the trend of social network spammer. © 2012 IEEE.

Online trendy topics detection in microblogs with selective user monitoring under cost constraints

As microblog services such as Twitter become a fast and convenient communication approach, identification of trendy topics in microblog services has great academic and business value. However detecting trendy topics is very challenging due to huge number of users and short-text posts in microblog diffusion networks. In this paper we introduce a trendy topics detection system under computation and communication resource constraints. In stark contrast to retrieving and processing the whole microblog contents, we develop an idea of selecting a small set of microblog users and processing their posts to achieve an overall acceptable trendy topic coverage, without exceeding resource budget for detection. We formulate the selection operation of these subset users as mixed-integer optimization problems, and develop heuristic algorithms to compute their approximate solutions. The proposed system is evaluated with real-time test data retrieved from Sina Weibo, the dominant microblog service provider in China. It's shown that by monitoring 500 out of 1.6 million microblog users and tracking their microposts (about 15,000 daily) with our system, nearly 65% trendy topics can be detected, while on average 5 hours earlier before they appear in Sina Weibo official trends.

A computational approach to measuring the correlation between expertise and social media influence for celebrities on microblogs

Existing approaches of social influence analysis usually focus on how to develop effective algorithms to quantize users' influence scores. They rarely consider a person's expertise levels which are arguably important to influence measures. In this paper, we propose a computational approach to measuring the correlation between expertise and social media influence, and we take a new perspective to understand social media influence by incorporating expertise into influence analysis. We carefully constructed a large dataset of 13,684 Chinese celebrities from Sina Weibo (literally 'Sina microblogging'). We found that there is a strong correlation between expertise levels and social media influence scores. In addition, different expertise levels showed influence variation patterns: high-expertise celebrities have stronger influence on the 'audience' in their expertise domains.

We know what you want to buy:a demographic-based system for product recommendation on microblogs

Product recommender systems are often deployed by e-commerce websites to improve user experience and increase sales. However, recommendation is limited by the product information hosted in those e-commerce sites and is only triggered when users are performing e-commerce activities. In this paper, we develop a novel product recommender system called METIS, a MErchanT Intelligence recommender System, which detects users' purchase intents from their microblogs in near real-time and makes product recommendation based on matching the users' demographic information extracted from their public profiles with product demographics learned from microblogs and online reviews. METIS distinguishes itself from traditional product recommender systems in the following aspects: 1) METIS was developed based on a microblogging service platform. As such, it is not limited by the information available in any specific e-commerce website. In addition, METIS is able to track users' purchase intents in near real-time and make recommendations accordingly. 2) In METIS, product recommendation is framed as a learning to rank problem. Users' characteristics extracted from their public profiles in microblogs and products' demographics learned from both online product reviews and microblogs are fed into learning to rank algorithms for product recommendation. We have evaluated our system in a large dataset crawled from Sina Weibo. The experimental results have verified the feasibility and effectiveness of our system. We have also made a demo version of our system publicly available and have implemented a live system which allows registered users to receive recommendations in real time. © 2014 ACM.