Role of electrostatic interactions for ligand recognition and specificity of peptide transporters.

BACKGROUND Peptide transporters are membrane proteins that mediate the cellular uptake of di- and tripeptides, and of peptidomimetic drugs such as β-lactam antibiotics, antiviral drugs and antineoplastic agents. In spite of their high physiological and pharmaceutical importance, the molecular recognition by these transporters of the amino acid side chains of short peptides and thus the mechanisms for substrate binding and specificity are far from being understood. RESULTS The X-ray crystal structure of the peptide transporter YePEPT from the bacterium Yersinia enterocolitica together with functional studies have unveiled the molecular bases for recognition, binding and specificity of dipeptides with a charged amino acid residue at the N-terminal position. In wild-type YePEPT, the significant specificity for the dipeptides Asp-Ala and Glu-Ala is defined by electrostatic interaction between the in the structure identified positively charged Lys314 and the negatively charged amino acid side chain of these dipeptides. Mutagenesis of Lys314 into the negatively charged residue Glu allowed tuning of the substrate specificity of YePEPT for the positively charged dipeptide Lys-Ala. Importantly, molecular insights acquired from the prokaryotic peptide transporter YePEPT combined with mutagenesis and functional uptake studies with human PEPT1 expressed in Xenopus oocytes also allowed tuning of human PEPT1's substrate specificity, thus improving our understanding of substrate recognition and specificity of this physiologically and pharmaceutically important peptide transporter. CONCLUSION This study provides the molecular bases for recognition, binding and specificity of peptide transporters for dipeptides with a charged amino acid residue at the N-terminal position.

RNAs and ribonucleoproteins in recognition and catalysis.

CONTENTS. 1. Did life begin with catalytic RNA?–2. Self-splicing and self-cleaving RNAs–2.1 Self-splicing of group I introns – 2.2 Self-splicing of group II introns – 2.3 Self-cleaving RNAs–3. Splicing mediated by trans-acting factors–3.1 Group III introns – 3.2 Splicing of nuclear pre-mRNAs – 3.3 Trans-splicing – 3.4 Is nuclear pre-mRNA splicing evolutionarily related to group I and group II self-splicing?– 3.5 Non-RNA mediated splicing of tRNAs–4. Processing of ribosomal precursor RNAs–5. Processing of pre-mRNA 3′ ends–5.1 Polyadenylation – 5.2 Histone pre-mRNA 3′ processing–6. Other RNPs involved in metabolic mechanisms–6.1 5′ end processing of pre-tRNAs by RNase P – 6.2 The signal recognition particle – 6.3 Telomerase – 6.4 RNA editing in trypanosomatid mitochondria–7. Why RNA?

Food Recognition for Dietary Assessment Using Deep Convolutional Neural Networks

Diet management is a key factor for the prevention and treatment of diet-related chronic diseases. Computer vision systems aim to provide automated food intake assessment using meal images. We propose a method for the recognition of already segmented food items in meal images. The method uses a 6-layer deep convolutional neural network to classify food image patches. For each food item, overlapping patches are extracted and classified and the class with the majority of votes is assigned to it. Experiments on a manually annotated dataset with 573 food items justified the choice of the involved components and proved the effectiveness of the proposed system yielding an overall accuracy of 84.9%.

The contributions of the hippocampal formation, perirhinal cortex and areas TH/TF to object, spatial and contextual recognition memory in primates

Adult monkeys (Macaca mulatta) with lesions of the hippocampal formation, perirhinal cortex, areas TH/TF, as well as controls were tested on tasks of object, spatial and contextual recognition memory. ^ Using a visual paired-comparison (VPC) task, all experimental groups showed a lack of object recognition relative to controls, although this impairment emerged at 10 sec with perirhinal lesions, 30 sec with areas TH/TF lesions and 60 sec with hippocampal lesions. In contrast, only perirhinal lesions impaired performance on delayed nonmatching-to-sample (DNMS), another task of object recognition memory. All groups were tested on DNMS with distraction (dDNMS) to examine whether the use of active cognitive strategies during the delay period could enable good performance on DNMS in spite of impaired recognition memory (revealed by the VPC task). Distractors affected performance of animals with perirhinal lesions at the 10-sec delay (the only delay in which their DNMS performance was above chance). They did not affect performance of animals with areas TH/TF lesions. Hippocampectomized animals were impaired at the 600-sec delay (the only delay at which prevention of active strategies would likely affect their behavior). ^ While lesions of areas TH/TF impaired spatial location memory and object-in-place memory, hippocampal lesions impaired only object-in-place memory. The pattern of results for perirhinal cortex lesions on the different task conditions indicated that this cortical area is not critical for spatial memory. ^ Finally, all three lesions impaired contextual recognition memory processes. The pattern of impairment appeared to result from the formation of only a global representation of the object and background, and suggests that all three areas are recruited for associating information across sources. ^ These results support the view that (1) the perirhinal cortex maintains storage of information about object and the context in which it is learned for a brief period of time, (2) areas TH/TF maintain information about spatial location and form associations between objects and their spatial relationship (a process that likely requires additional time) and (3) the hippocampal formation mediates associations between objects, their spatial relationship and the general context in which these associations are formed (an integrative function that requires additional time). ^

Roles of mismatch repair proteins in the recognition and processing of DNA interstrand crosslinks in human cells

DNA interstrand crosslinks (ICLs) are among the most toxic type of damage to a cell. Many ICL-inducing agents are widely used as therapeutic agents, e.g. cisplatin, psoralen. A bettor understanding of the cellular mechanism that eliminates ICLs is important for the improvement of human health. However, ICL repair is still poorly understood in mammals. Using a triplex-directed site-specific ICL model, we studied the roles of mismatch repair (MMR) proteins in ICL repair in human cells. We are also interested in using psoralen-conjugated triplex-forming oligonucleotides (TFOs) to direct ICLs to a specific site in targeted DNA and in the mammalian genomes. ^ MSH2 protein is the common subunit of two MMR recognition complexes, and MutSα and MutSβ. We showed that MSH2 deficiency renders human cell hypersensitive to psoralen ICLs. MMR recognition complexes bind specifically to triplex-directed psoralen ICLs in vitro. Together with the fact that psoralen ICL-induced repair synthesis is dramatically decreased in MSH2 deficient cell extracts, we demonstrated that MSH2 function is critical for the recognition and processing of psoralen ICLs in human cells. Interestingly, lack of MSH2 does not reduce the level of psoralen ICL-induced mutagenesis in human cells, suggesting that MSH2 does not contribute to error-generating repair of psoralen ICLs, and therefore, may represent a novel error-free mechanism for repairing ICLs. We also studied the role of MLH1, anther key protein in MMR, in the processing of psoralen ICLs. MLH1-deficient human cells are more resistant to psoralen plus UVA treatment. Importantly, MLH1 function is not required for the mutagenic repair of psoralen ICLs, suggesting that it is not involved in the error-generating repair of this type of DNA damage in human cells. ^ These are the first data indicating mismatch repair proteins may participate in a relatively error-free mechanism for processing psoralen ICL in human cells. Enhancement of MMR protein function relative to nucleotide excision repair proteins may reduce the mutagenesis caused by DNA ICLs in humans. ^ In order to specifically target ICLs to mammalian genes, we identified novel TFO target sequences in mouse and human genomes. Using this information, many critical mammalian genes can now be targeted by TFOs.^

Searching for the ideal DNA recognition code of covalent ligands

The combitiatorial approach restriction endonuclease protection selection and amplification REPSA was successfully used to determine ideal DNA interactions sites of covalent ligands. Unlike most other combinatorial methods, REPSA is based on inhibition of enzymatic cleavage by specific ligand-DNA complexes, which enables identification of binding sites of various ligands. However, the inherent nature of this technique posses a problem during selection of binding sites of covalent ligands. By modifying the technique according to the nature of the ligand, we demonstrate the flexibility of REPSA in identifying the preferred binding sites for monocovalent ligands, topoisomerase I and tallimustine, and the bicovalent ligand topoisomerase II. From among the preferred binding sites, we identified the consensus binding sequence of camptothecin induced topoisomerase I cleavage as ‘aGWT/Gc’, and tallimustine consensus sequences as ‘GTTCTA’ and ‘TTTTTTC’. We have shown for the first time that preferential binding of tallimustine occurs at sequences not previously reported. Furthermore, our data indicate that tallimustine is a novel DNA minor groove, guanine-specific alkylating agent. ^ Additionally, we have demonstrated in vivo that sequence-specific covalent DNA-binding small molecules have the ability to regulate transcription by inhibiting RNA polymerase II. Tallimustine, binding to its preferred sequences located in the 5′ untranslated region were an effective impediment for transcribing polymerase II. The ability of covalent binding small molecules to target predetermined DNA sequences located downstream of the promoter suggests a general approach for regulation of gene expression. ^

Turbidite recognition and radiocarbon ages of marine sediment cores along the continental margin of Chile

Much progress has been made in estimating recurrence intervals of great and giant subduction earthquakes using terrestrial, lacustrine, and marine paleoseismic archives. Recent detailed records suggest these earthquakes may have variable recurrence periods and magnitudes forming supercycles. Understanding seismic supercycles requires long paleoseismic archives that record timing and magnitude of such events. Turbidite paleoseismic archives may potentially extend past earthquake records to the Pleistocene and can thus complement commonly shorter-term terrestrial archives. However, in order to unambiguously establish recurring seismicity as a trigger mechanism for turbidity currents, synchronous deposition of turbidites in widely spaced, isolated depocenters has to be ascertained. Furthermore, characteristics that predispose a seismically active continental margin to turbidite paleoseismology and the correct sample site selection have to be taken into account. Here we analyze 8 marine sediment cores along 950 km of the Chile margin to test for the feasibility of compiling detailed and continuous paleoseismic records based on turbidites. Our results suggest that the deposition of areally widespread, synchronous turbidites triggered by seismicity is largely controlled by sediment supply and, hence, the climatic and geomorphic conditions of the adjacent subaerial setting. The feasibility of compiling a turbidite paleoseismic record depends on the delicate balance between sufficient sediment supply providing material to fail frequently during seismic shaking and sufficiently low sedimentation rates to allow for coeval accumulation of planktonic foraminifera for high-resolution radiocarbon dating. We conclude that offshore northern central Chile (29-32.5°S) Holocene turbidite paleoseismology is not feasible, because sediment supply from the semi-arid mainland is low and almost no Holocene turbidity-current deposits are found in the cores. In contrast, in the humid region between 36 and 38°S frequent Holocene turbidite deposition may generally correspond to paleoseismic events. However, high terrigenous sedimentation rates prevent high-resolution radiocarbon dating. The climatic transition region between 32.5 and 36°S appears to be best suited for turbidite paleoseismology.

Automated Tools for Layer Recognition (BMPix) and Counting (PEAK)

We present tools for rapid and quantitative detection of sediment lamination. The BMPix tool extracts color and gray-scale curves from images at pixel resolution. The PEAK tool uses the gray-scale curve and performs, for the first time, fully automated counting of laminae based on three methods. The maximum count algorithm counts every bright peak of a couplet of two laminae (annual resolution) in a smoothed curve. The zero-crossing algorithm counts every positive and negative halfway-passage of the curve through a wide moving average, separating the record into bright and dark intervals (seasonal resolution). The same is true for the frequency truncation method, which uses Fourier transformation to decompose the curve into its frequency components before counting positive and negative passages. We applied the new methods successfully to tree rings, to well-dated and already manually counted marine varves from Saanich Inlet, and to marine laminae from the Antarctic continental margin. In combination with AMS14C dating, we found convincing evidence that laminations in Weddell Sea sites represent varves, deposited continuously over several millennia during the last glacial maximum. The new tools offer several advantages over previous methods. The counting procedures are based on a moving average generated from gray-scale curves instead of manual counting. Hence, results are highly objective and rely on reproducible mathematical criteria. Also, the PEAK tool measures the thickness of each year or season. Since all information required is displayed graphically, interactive optimization of the counting algorithms can be achieved quickly and conveniently.

The gap between recognition and the 'compensation business' : the claim against Britain for compensation by Kenya's former Mau Mau fighters

During the first Kibaki administration (2002-2007), a movement by the former Mau Mau fighters demanded recognition for the role that they had played in the achievement of independence. They began to demand, also, monetary compensation for past injustices. Why had it taken over 40 years (from independence in 1963) for the former Mau Mau fighters to initiate this movement? What can be observed as the outcome of their movement? To answer these questions, three different historical currents need to be taken into account. These were, respectively, changing trends in the government of Kenya, progress in historical research into the actual circumstances of colonial control, and a realization, based on mounting experience, that launching a legal action against Britain could turn out to be a lucrative initiative. This paper concludes that, regardless of the actual purpose of the legal case, neither of their objectives was certain to be achieved. Two inescapable realities remain: the doubts cast on the reputation of the government by its decision to lift the Mau Mau‟s outlaw status – a decision that was widely seen as a latter-day example of the „Kikuyu favouritism‟ policy followed by the first Kibaki administration – and the popular interpretation of the involvement of Leigh Day, well known in Kenya ever since the unexploded bombs case for its success in obtaining substantial compensation payments, as a vehicle for squeezing large amounts of money from the British government for the benefit of the Kikuyu people.

Enhancing Activity Recognition by Fusing Inertial and Biometric Information

Activity recognition is an active research field nowadays, as it enables the development of highly adaptive applications, e.g. in the field of personal health. In this paper, a light high-level fusion algorithm to detect the activity that an individual is performing is presented. The algorithm relies on data gathered from accelerometers placed on different parts of the body, and on biometric sensors. Inertial sensors allow detecting activity by analyzing signal features such as amplitude or peaks. In addition, there is a relationship between the activity intensity and biometric response, which can be considered together with acceleration data to improve the accuracy of activity detection. The proposed algorithm is designed to work with minimum computational cost, being ready to run in a mobile device as part of a context-aware application. In order to enable different user scenarios, the algorithm offers best-effort activity estimation: its quality of estimation depends on the position and number of the available inertial sensors, and also on the presence of biometric information.

Area-delay Trade-offs of Texture Decompressors for a Graphics Processing Unit

Graphics Processing Units have become a booster for the microelectronics industry. However, due to intellectual property issues, there is a serious lack of information on implementation details of the hardware architecture that is behind GPUs. For instance, the way texture is handled and decompressed in a GPU to reduce bandwidth usage has never been dealt with in depth from a hardware point of view. This work addresses a comparative study on the hardware implementation of different texture decompression algorithms for both conventional (PCs and video game consoles) and mobile platforms. Circuit synthesis is performed targeting both a reconfigurable hardware platform and a 90nm standard cell library. Area-delay trade-offs have been extensively analyzed, which allows us to compare the complexity of decompressors and thus determine suitability of algorithms for systems with limited hardware resources.