Deciphering complex patterns of class-I HLA-peptide cross-reactivity via hierarchical grouping

T-cell responses in humans are initiated by the binding of a peptide antigen to a human leukocyte antigen (HLA) molecule. The peptide-HLA complex then recruits an appropriate T cell, leading to cell-mediated immunity. More than 2000 HLA class-I alleles are known in humans, and they vary only in their peptide-binding grooves. The polymorphism they exhibit enables them to bind a wide range of peptide antigens from diverse sources. HLA molecules and peptides present a complex molecular recognition pattern, as many peptides bind to a given allele and a given peptide can be recognized by many alleles. A powerful grouping scheme that not only provides an insightful classification, but is also capable of dissecting the physicochemical basis of recognition specificity is necessary to address this complexity. We present a hierarchical classification of 2010 class-I alleles by using a systematic divisive clustering method. All-pair distances of alleles were obtained by comparing binding pockets in the structural models. By varying the similarity thresholds, a multilevel classification was obtained, with 7 supergroups, each further subclassifying to yield 72 groups. An independent clustering performed based only on similarities in their epitope pools correlated highly with pocket-based clustering. Physicochemical feature combinations that best explain the basis of clustering are identified. Mutual information calculated for the set of peptide ligands enables identification of binding site residues contributing to peptide specificity. The grouping of HLA molecules achieved here will be useful for rational vaccine design, understanding disease susceptibilities and predicting risk of organ transplants.

Microwave spectroscopic and theoretical investigations of the strongly hydrogen bonded hexafluoroisopropanol...water complex

This paper reports microwave spectroscopic and theoretical investigations on the interaction of water with hexafluoroisopropanol (HFIP). The HFIP monomer can exist in two conformations, antiperiplanar (AP) and synclinical (SC). The former is about 5 kJ mol(-1) more stable than the latter. Theoretical calculations predicted three potential minima for the complex, two having AP and one having SC conformations. Though, the binding energy for the HFIP(SC)...H2O turned out to be larger than that for the other two conformers having HFIP in the AP form, the global minimum for the complex in the potential energy hypersurface had HFIP in the AP form. Experimental rotational constants for four isotopologues measured using a pulsed nozzle Fourier transform microwave spectrometer, correspond to the global minimum in the potential energy hypersurface. The structural parameters and the internal dynamics of the complex could be determined from the rotational spectra of the four isotopologues. The global minimum has the HFIP(AP) as a hydrogen bond donor forming a strong hydrogen bond with H2O. To characterize the strength of the bonding and to probe the other interactions within the complex, atoms in molecules, non-covalent interaction index and natural bond orbital theoretical analyses have been performed.

HuR Displaces Polypyrimidine Tract Binding Protein To Facilitate La Binding to the 3 ` Untranslated Region and Enhances Hepatitis C Virus Replication

HuR is a ubiquitous, RNA binding protein that influences the stability and translation of several cellular mRNAs. Here, we report a novel role for HuR, as a regulator of proteins assembling at the 3' untranslated region (UTR) of viral RNA in the context of hepatitis C virus (HCV) infection. HuR relocalizes from the nucleus to the cytoplasm upon HCV infection, interacts with the viral polymerase (NS5B), and gets redistributed into compartments of viral RNA synthesis. Depletion in HuR levels leads to a significant reduction in viral RNA synthesis. We further demonstrate that the interaction of HuR with the 3' UTR of the viral RNA affects the interaction of two host proteins, La and polypyrimidine tract binding protein (PTB), at this site. HuR interacts with La and facilitates La binding to the 3' UTR, enhancing La-mediated circularization of the HCV genome and thus viral replication. In addition, it competes with PTB for association with the 3' UTR, which might stimulate viral replication. Results suggest that HuR influences the formation of a cellular/viral ribonucleoprotein complex, which is important for efficient initiation of viral RNA replication. Our study unravels a novel strategy of regulation of HCV replication through an interplay of host and viral proteins, orchestrated by HuR. IMPORTANCE Hepatitis C virus (HCV) is highly dependent on various host factors for efficient replication of the viral RNA. Here, we have shown how a host factor (HuR) migrates from the nucleus to the cytoplasm and gets recruited in the protein complex assembling at the 3' untranslated region (UTR) of HCV RNA. At the 3' UTR, it facilitates circularization of the viral genome through interaction with another host factor, La, which is critical for replication. Also, it competes with the host protein PTB, which is a negative regulator of viral replication. Results demonstrate a unique strategy of regulation of HCV replication by a host protein through alteration of its subcellular localization and interacting partners. The study has advanced our knowledge of the molecular mechanism of HCV replication and unraveled the complex interplay between the host factors and viral RNA that could be targeted for therapeutic interventions.

Visible light-induced cytotoxicity of a dinuclear iron(III) complex of curcumin with low-micromolar IC50 value in cancer cells

Photoactive metal complexes have emerged as potential candidates in the photodynamic therapy (PDT) of cancer. We present here the synthesis, characterization and visible light-triggered anticancer activity of two novel mixed-ligand oxo-bridged iron(III) complexes, viz., {Fe(L)(acac)}(2)(mu-O)](ClO4)(2) (1) and {Fe (L)(cur)}(2)(mu-O)](ClO4)(2) (2) where L is bis-(2-pyridylmethyl)-benzylamine, acac is acetylacetonate and cur is the monoanion of curcumin (bis(4-hydroxy-3-methoxyphenyl)-1,6-diene-3,5-dione). The crystal structure of complex 1 (as PF6 salt, 1a) shows distorted octahedral geometry of each iron(III) centre formed by the FeN3O3 core. The 1: 2 electrolytic complexes are stable in solution and retain their oxo-bridged identity in aqueous medium. Complex 2 has a strong absorption band in the visible region and shows promising photocytotoxicity in HeLa and MCF-7 cancer cells in visible light giving respective IC50 values of 3.1 +/- 0.4 lM and 4.9 +/- 0.5 lM while remains non-toxic in the dark (IC50 > 50 lM). The control complex 1 is inactive both in the light and dark. Complex 2 accumulates in cytoplasm of HeLa and MCF-7 cells as evidenced from fluorescence microscopy and triggers apoptotic cell death via light-assisted generation of reactive oxygen species (ROS). Taken together, complex 2 with its promising photocytotoxicity but negligible dark toxicity in cancer cells has significant photochemotherapeutic potential for applications in PDT. (C) 2015 Elsevier B.V. All rights reserved.

Mitochondrial selectivity and remarkable photocytotoxicity of a ferrocenyl neodymium(III) complex of terpyridine and curcumin in cancer cells

A series of four novel neodymium(III) complexes of the formulation Nd(R-tpy)(O-O)(NO3)(2)] (1-4), where R-tpy is 4'-phenyl-2,2': 6', 2''-terpyridine (Ph-tpy; 1, 2) and 4'-ferrocenyl-2,2': 6', 2''-terpyridine (Fc-tpy; 3, 4); O-O is the conjugate base of acetylacetone (Hacac; 1, 3) or curcumin (Hcurc; 2, 4), are synthesized and characterized. The single crystal structure of 1 shows that the complex is a discrete mononuclear species with the Nd(III) centre in a nine coordinate environment provided by a set of O6N3 donor atoms. Complexes 1 and 3 having the simple acac ligand are prepared as control compounds. Complex 4, possessing an appended ferrocenyl (Fc) and the curcumin moiety, is remarkably photocytotoxic to HeLa and MCF-7 cancer cells in visible light giving respective IC50 values of 0.7 mu M and 2.1 mu M while being significantly less toxic to MCF-10A normal cells (IC50 = 34 mu M) and in the dark (IC50 > 50 mu M). The phenyl appended complex 2, lacking a ferrocenyl moiety, is significantly less toxic to both the cell lines when compared with 4. Complexes 1 and 3, lacking the photoactive curcumin moiety, do not show any apparent toxicity both in light and in the dark. The cell death is apoptotic in nature and is mediated by the light-induced formation of reactive oxygen species (ROS). Fluorescence imaging experiment with HeLa cells reveals mitochondrial accumulation of complex 4 within 4 h of incubation. The complexes bind to calf thymus (ct) DNA with moderate affinity giving K-b values in the range of 10(4)-10(5) M-1. The curcumin complexes 2 and 4 cleave plasmid supercoiled DNA to its nicked circular form in visible light via O-1(2) and (OH)-O-center dot pathways. The presence of the ferrocenyl moiety is likely to be responsible for the enhanced cellular uptake and photocytotoxicity of complex 4. Thus, the mitochondria targeting complex 4, being remarkably cytotoxic in light but non-toxic in the dark and to normal cells, is a potential candidate for photochemotherapeutic applications.

Complex stress functions and plastic zone sizes for notch cracks subjected to various loading conditions

In this paper, the conformal mapping method is used to solve the plane problem of an infinite plate containing a central lip-shaped notch subjected to biaxial loading at a remote boundary or a surface uniform pressure on the notch. The stress intensity factors KI and KII are obtained by the derived complex stress functions. The simple analytical expressions can be applied to the situation of cracks originating from a circular or an elliptical notch. The plastic zone sizes for such notch cracks are subsequently evaluated in light of the Dugdale strip yield concept. The results are consistent with available numerical data.

IP Simulation of Micro Gas Flows under 3-D Head Sliders

Gas film lubrication of a three-dimensional flat read-write head slider is calculated using the information preservation (IP) method and the direct simulation Monte Carlo (DSMC) method, respectively. The pressure distributions on the head slider surface at different velocities and flying heights obtained by the two methods are in excellent agreement. IP method is also employed to deal with head slider with three-dimensional complex configuration. The pressure distribution on the head slider surface and the net lifting force obtained by the IP method also agree well with those of DSMC method. Much less (of the order about 10(2) less) computational time (the sum of the time used to reach a steady stage and the time used in sampling process) is needed by the IP method than the DSMC method and such an advantage is more remarkable as the gas velocity decreases.

An Integrated Assessment of the Introduction of Lionfish (Pterois volitans/miles complex) to the Western Atlantic Ocean.

Lionfish (Pterois volitans/miles complex) are venomous coral reef fishes from the Indian and western Pacific oceans that are now found in the western Atlantic Ocean. Adult lionfish have been observed from Miami, Florida to Cape Hatteras, North Carolina, and juvenile lionfish have been observed off North Carolina, New York, and Bermuda. The large number of adults observed and the occurrence of juveniles indicate that lionfish are established and reproducing along the southeast United States coast. Introductions of marine species occur in many ways. Ballast water discharge, a very common method of introduction for marine invertebrates, is responsible for many freshwater fish introductions. In contrast, most marine fish introductions result from intentional stocking for fishery purposes. Lionfish, however, likely were introduced via unintentional or intentional aquarium releases, and the introduction of lionfish into United States waters should lead to an assessment of the threat posed by the aquarium trade as a vector for fish introductions. Currently, no management actions are being taken to limit the effect of lionfish on the southeast United States continental shelf ecosystem. Further, only limited funds have been made available for research. Nevertheless, the extent of the introduction has been documented and a forecast of the maximum potential spread of lionfish is being developed. Under a scenario of no management actions and limited research, three predictions are made: ● With no action, the lionfish population will continue to grow along the southeast United States shelf. ● Effects on the marine ecosystem of the southeast United States will become more noticeable as the lionfish population grows. ● There will be incidents of lionfish envenomations of divers and/or fishers along the east coast of the United States. Removing lionfish from the southeast United States continental shelf ecosystem would be expensive and likely impossible. A bounty could be established that would encourage the removal of fish and provide specimens for research. However, the bounty would need to be lower than the price of fish in the aquarium trade (~$25-$50 each) to ensure that captured specimens were from the wild. Such a low bounty may not provide enough incentive for capturing lionfish in the wild. Further, such action would only increase the interaction between the public and lionfish, increasing the risk of lionfish envenomations. As the introduction of lionfish is very likely irreversible, future actions should focus on five areas. 1) The population of lionfish should be tracked. 2) Research should be conducted so that scientists can make better predictions regarding the status of the invasion and the effects on native species, ecosystem function, and ecosystem services. 3) Outreach and education efforts must be increased, both specifically toward lionfish and more generally toward the aquarium trade as a method of fish introductions. 4) Additional regulation should be considered to reduce the frequency of marine fish introduction into U.S. waters. However, the issue is more complicated than simply limiting the import of non-native species, and these complexities need to be considered simultaneously. 5) Health care providers along the east coast of the United States need to be notified that a venomous fish is now resident along the southeast United States. The introduction and spread of lionfish illustrates the difficulty inherent in managing introduced species in marine systems. Introduced species often spread via natural mechanisms after the initial introduction. Efforts to control the introduction of marine fish will fail if managers do not consider the natural dispersal of a species following an introduction. Thus, management strategies limiting marine fish introductions need to be applied over the scale of natural ecological dispersal to be effective, pointing to the need for a regional management approach defined by natural processes not by political boundaries. The introduction and success of lionfish along the east coast should change the long-held perception that marine fish invasions are a minimal threat to marine ecosystems. Research is needed to determine the effects of specific invasive fish species in specific ecosystems. More broadly, a cohesive plan is needed to manage, mitigate and minimize the effects of marine invasive fish species on ecosystems that are already compromised by other human activities. Presently, the magnitude of marine fish introductions as a stressor on marine ecosystems cannot be quantified, but can no longer be dismissed as negligible. (PDF contains 31 pages)

Modeling smart grids as complex systems through the implementation of intelligent hubs

ICINCO 2010