Rapid mass spectrometric determination of disulfide connectivity in peptides and proteins

Disulfide crosslinks are ubiquitous in natural peptides and proteins, providing rigidity to polypeptide scaffolds. The assignment of disulfide connectivity in multiple crosslinked systems is often difficult to achieve. Here, we show that rapid unambiguous characterisation of disulfide connectivity can be achieved through direct mass spectrometric CID fragmentation of the disulfide intact polypeptides. The method requires a direct mass spectrometric fragmentation of the native disulfide bonded polypeptides and subsequent analysis using a newly developed program, DisConnect. Technical difficulties involving direct fragmentation of proteins are surmounted by an initial proteolytic nick and subsequent determination of the structures of these proteolytic peptides through DisConnect. While the connectivity in proteolytic fragments containing one cystine is evident from the MS profile alone, those with multiple cystines are subjected to subsequent mass spectrometric fragmentation. The wide applicability of this method is illustrated using examples of peptide hormones, peptide toxins, proteins, and disulfide foldamers of a synthetic analogue of a marine peptide toxin. The method, coupled with DisConnect, provides an unambiguous, straightforward approach, especially useful for the rapid screening of the disulfide crosslink fidelity in recombinant proteins, determination of disulfide linkages in natural peptide toxins and characterization of folding intermediates encountered in oxidative folding pathways.

Detection of Hepatitis B DNA Sequences on Polyelectrolyte Based Non-Covalently Functionalized Flexible Plastic Substrates

Development of simple functionalization methods to attach biomolecules such as proteins and DNA on inexpensive substrates is important for widespread use of low cost, disposable biosensors. Here, we describe a method based on polyelectrolyte multilayers to attach single stranded DNA molecules to conventional glass slides as well as a completely non-standard substrate, namely flexible plastic transparency sheets. We then use the functionalized transparency sheets to specifically detect single stranded Hepatitis B DNA sequences from samples. We also demonstrate a blocking method for reducing non-specific binding of target DNA sequences using negatively charged polyelectrolyte molecules. The polyelectrolyte based functionalization method, which relies on surface charge as opposed to covalent surface linkages, could be an attractive platform to develop assays on inexpensive substrates for low cost biosensing.

Cross-Linked, Biodegradable, Cytocompatible Salicylic Acid Based Polyesters for Localized, Sustained Delivery of Salicylic Acid: An In Vitro Study

In order to suppress chronic inflammation while supporting cell proliferation, there has been a continuous surge toward development of polymers with the intention of delivering anti-inflammatory molecules in a sustained manner. In the above backdrop, we report the synthesis of a novel, stable, cross-linked polyester with salicylic acid (SA) incorporated in the polymeric backbone and propose a simple synthesis route by melt condensation. The as-synthesized polymer was hydrophobic with a glass transition temperature of 1 degrees C, which increases to 17 degrees C upon curing. The combination of NMR and FT-IR spectral techniques established the ester linkages in the as-synthesized SA-based polyester. The pH-dependent degradation rate and the rate of release of salicylic acid from the as-synthesized SA-based polymer were studied at physiological conditions in vitro. The polyester underwent surface erosion and exhibited linear degradation kinetics in which a change in degradation rate is observed after 4-10 days and 24% mass loss was recorded after 4 months at 37 degrees C and pH 7.4. The delivery of salicylic acid also showed a similar change in slopes, with a sustained release rate of 3.5% in 4 months. The cytocompatibility studies of these polyesters were carried out with C2C12 murine myoblast cells using techniques like MTT assay and flow cytometry. Our results strongly suggest that SA-based polyester supports cell proliferation for 3 days in culture and do not cause cell death (<7%), as quantified by propidium iodide (PI) stained cells. Hence, these polyesters can be used as implant materials for localized, sustained delivery of salicylic acid and have applications in adjuvant cancer therapy, chronic wound healing, and as an alternative to commercially available polymers like poly(lactic acid) and poly(glycolic acid) or their copolymers.

Amide Functionalized Microporous Organic Polymer (Am-MOP) for Selective CO2 Sorption and Catalysis

We report the design and synthesis of an amide functionalized microporous organic polymer (Am-MOP) prepared from trimesic acid and p-phenylenediamine using thionyl chloride as a reagent. Polar amide (CONH) functional groups act as a linking unit between the node and spacer and constitute the pore wall of the continuous polymeric network. The strong covalent bonds between the building blocks (trimesic acid and p-phenylenediamine) through amide bond linkages provide high thermal and chemical stability to Am-MOP. The presence of a highly polar pore surface allows selective CO2 uptake at 195 K over other gases such as N-2, Ar, and O-2. The CO2 molecule interacts with amide functional groups via Lewis acid base type interactions as demonstrated through DFT calculations. Furthermore, for the first time Am-MOP with basic functional groups has been exploited for the Knoevenagel condensation reaction between aldehydes and active methylene compounds. Availability of a large number of catalytic sites per volume and confined microporosity gives enhanced catalytic efficiency and high selectivity for small substrate molecules.

Influence of anthropogenic contamination on fluoride concentration in groundwater: A study of Mulbagal town, Kolar district, Karnataka

Groundwater contamination is a serious concern in India. Major geogenic contaminants include fluoride, arsenic and iron, while common anthropogenic contaminants include nitrate, metals, organics and microbial contamination. Besides, known point and diffuse sources, groundwater c ontamination from inf iltration of pit to ilet leachate is an emerging concern. The study area of this paper is Kolar district in Karnataka that is hot spot of fluoride contamination. The absence of fluoride contamination in Mulbagal town and the alterations in groundwater chemistry from infiltration of pit toilet leachate motivated the author to examine the possible linkages between anthropogenic contamination and fluoride concentration in groundwater of Mulbagal town. Analysis of the groundwater chemistry revealed that the groundwater in Mulbagal town is under saturated with respect to calcite that suppresses the dissolution of fluorite and the fluoride concentration in the groundwater. The slightly acidic pH of the groundwater is considered responsible to facilitate calcite dissolution under saturation.

GHG footprint of major cities in India

Concentration of greenhouse gases (GHG) in the atmosphere has been increasing rapidly during the last century due to ever increasing anthropogenic activities resulting in significant increases in the temperature of the Earth causing global warming. Major sources of GHG are forests (due to human induced land cover changes leading to deforestation), power generation (burning of fossil fuels), transportation (burning fossil fuel), agriculture (livestock, farming, rice cultivation and burning of crop residues), water bodies (wetlands), industry and urban activities (building, construction, transport, solid and liquid waste). Aggregation of GHG (CO2 and non-CO2 gases), in terms of Carbon dioxide equivalent (CO(2)e), indicate the GHG footprint. GHG footprint is thus a measure of the impact of human activities on the environment in terms of the amount of greenhouse gases produced. This study focuses on accounting of the amount of three important greenhouses gases namely carbon dioxide (CO2), methane (CH4) and nitrous oxide (N2O) and thereby developing GHG footprint of the major cities in India. National GHG inventories have been used for quantification of sector-wise greenhouse gas emissions. Country specific emission factors are used where all the emission factors are available. Default emission factors from IPCC guidelines are used when there are no country specific emission factors. Emission of each greenhouse gas is estimated by multiplying fuel consumption by the corresponding emission factor. The current study estimates GHG footprint or GHG emissions (in terms of CO2 equivalent) for Indian major cities and explores the linkages with the population and GDP. GHG footprint (Aggregation of Carbon dioxide equivalent emissions of GHG's) of Delhi, Greater Mumbai, Kolkata, Chennai, Greater Bangalore, Hyderabad and Ahmedabad are found to be 38,633.2 Gg, 22,783.08 Gg, 14,812.10 Gg, 22,090.55 Gg, 19,796.5 Gg, 13,734.59 Gg and 91,24.45 Gg CO2 eq., respectively. The major contributors sectors are transportation sector (contributing 32%, 17.4%, 13.3%, 19.5%, 43.5%, 56.86% and 25%), domestic sector (contributing 30.26%, 37.2%, 42.78%, 39%, 21.6%, 17.05% and 27.9%) and industrial sector (contributing 7.9%, 7.9%, 17.66%, 20.25%, 1231%, 11.38% and 22.41%) of the total emissions in Delhi, Greater Mumbai, Kolkata, Chennai, Greater Bangalore, Hyderabad and Ahmedabad, respectively. Chennai emits 4.79 t of CO2 equivalent emissions per capita, the highest among all the cities followed by Kolkata which emits 3.29 t of CO2 equivalent emissions per capita. Also Chennai emits the highest CO2 equivalent emissions per GDP (2.55 t CO2 eq./Lakh Rs.) followed by Greater Bangalore which emits 2.18 t CO2 eq./Lakh Rs. (C) 2015 Elsevier Ltd. All rights reserved.

Mechano-Chemical Coupling at Molecular Level: Forced Dissociation of Selectin/Ligand Binding

Mechano-chemical coupling is a common phenomenon that exists in various biological processes at different physiological levels. Bone tissue remodeling strongly depends on the local mechanical load. Leukocytes are sheared to form the transient aggregates with platelets or other leukocytes in the circulation. Flow pattern affects the signal transduction pathways in endothelial cells. Receptor/ligand interactions are important to cell adhesion since they supply the physical linkages...

Structure-flexoelastic properties of bimesogenic liquid crystals

In this paper, we report on the flexoelastic and viscoelastic ratios for a number of bimesogens compounds with the same generic structure. Values are obtained indirectly by measuring the flexoelectro-optic response in the chiral nematic phase. By varying the molecular structure we alter the bend angle, transverse dipole moment, and length of the molecule. First, to examine the influence of the bend angle we use a homologous series whereby the only alteration in the molecular structure is the number of methylene units in the aliphatic spacer, n. Results show that the flexoelastic ratio, e K, and the effective flexoelectric coefficient, e, both exhibit an odd-even effect with values for n=odd being greater than that for n=even. This is understood in terms of an increase in the bend angle of the molecule and an increase in the transverse dipole moment. Second, in order to investigate the impact of the dipole moment, we have altered the mesogenic units so as to vary the longitudinal dipole moment and used different linkages in the aliphatic spacer in an attempt to alter the transverse dipole moment. Qualitatively, the results demonstrate that the odd-spaced bimesogen with larger transverse dipole moments exhibit larger flexoelastic ratios. © 2007 The American Physical Society.

PICES-GLOBEC International Program on Climate Change and Carrying Capacity

Introduction [pdf, 0.27 MB] Methods [pdf, 0.15 MB] Results and discussion [pdf, 2.1 MB] Conclusions [pdf, 0.12 MB] Appendix A: Data gathering review, results and balancing [pdf, 0.3 MB] Appendix B: Data tables [pdf, 0.35 MB] Appendix C: BASS Workshop on the "Development of a conceptual model of the subarctic Pacific Basin ecosystems" [pdf, 0.16 MB] Appendix D: BASS/MODEL Workshop on "Higher trohic level modeling" [pdf, 0.24 MB] Appendix E: BASS/MODEL Workshop to review ecosystem models for the subarctic Pacific gyres [pdf, 4.39 MB] Appendix F: BASS/MODEL Workshop on "Perturbation analysis" on subarctic Pacific gyre ecosystem models using ECOPATH/ECOSIM" [pdf, 0.37 MB] Appendix G: Proposal for a BASS Workshop on "Linkages between open and coastal systems" [pdf, 0.15 MB] References [pdf, 0.14 MB] (97 page document)

Charting multidisciplinary research and action priorities towards the conservation and sustainable management of sea turtles in the Pacific Ocean: a focus on Malaysia

The four sea turtle species found in Malaysia are the leatherback, olive ridley, green and hawksbill. The threats to these species are acute. Populations of leatherback, olive ridley and hawksbill turtles are on the brink of collapse – threatening a biodiversity crisis in Malaysia and the region. This proceedings contains 8 technical papers presented at a workshop convened in Kijal, Terengganu to chart new directions in the conservation of Malaysia's critically endangered sea turtles and to reverse population decline. They represent a wide range of issues from aspects of biology to a review of 40 years of sea turtle conservation. A paper on the socioeconomic linkages and impacts of fisheries was also included as the workshop adopted a multidisciplinary approach to address the issues. Two case studies, including successful restoration examples from international experiences and restoration efforts in Sabah, pave the way for enhancing turtle conservation in the country.

Peces continentales de la Argentina: Bibliografía. Gymnocharacinus bergii.

Espanol: Este trabajo tiene como objetivo la recopilación de la información existente sobre las especies presentes en nuestro territorio. Para ello, se elaboraron fichas que contienen las referencias argentinas y de aquellas revisiones de autores extranjeros que involucran taxas de nuestro país. Como principales fuentes de información se consultaron Ringuelet et al. (1967), CLOFFSCA (2003), López et al. (2003; 2006), Liotta (2006) y las bases de datos on-line de W. N. Eschmeyer y Fish Base. Esta es una publicación abierta, por lo que requerirá de actualización permanente, para lo cual sería necesario se sume a nuestra tarea la buena voluntad y colaboración de la comunidad ictiológica. Esperamos con este aporte colaborar en la construcción de una base de datos de los peces continentales de la Argentina. La concreción de esta herramienta dará soporte a los planes generados desde los diferentes sectores involucrados en el tema, además de fomentar la difusión de la disciplina y la reafirmación de los derechos soberanos sobre nuestros recursos naturales. English: The goal of this work is to compile the existing information about the species present in our country. This has been done by means of datasheets that list both local references and reviews by foreign authors that include Argentine taxa. The main information sources consulted were Ringuelet et al. (1967), CLOFFSCA (2003), López et al. (2003; 2006), Liotta (2006) and the online databases of W. N. Eschmeyer and Fish Base. This is an open-ended publication. As such, it will require permanent updating, which will depend on the good will and collaboration of the ichthyological community. With this contribution, we hope to help to build a database of the freshwater fish of Argentina. The completion of this project will provide support to diverse plans emanating from different sectors, as well as promote the diffusion of this field of research and reaffirm Argentina’s sovereign rights over our natural resources. (Texto en Espanol Y English. PDF tiene nueve paginas.)

Peces continentales de la Argentina: Bibliografía. Lepidosiren paradoxa.

Espanol: Este trabajo tiene como objetivo la recopilación de la información existente sobre las especies presentes en nuestro territorio. Para ello, se elaboraron fichas que contienen las referencias argentinas y de aquellas revisiones de autores extranjeros que involucran taxas de nuestro país. Como principales fuentes de información se consultaron Ringuelet et al. (1967), CLOFFSCA (2003), López et al. (2003; 2006), Liotta (2006) y las bases de datos on-line de W. N. Eschmeyer y Fish Base. Esta es una publicación abierta, por lo que requerirá de actualización permanente, para lo cual sería necesario se sume a nuestra tarea la buena voluntad y colaboración de la comunidad ictiológica. Esperamos con este aporte colaborar en la construcción de una base de datos de los peces continentales de la Argentina. La concreción de esta herramienta dará soporte a los planes generados desde los diferentes sectores involucrados en el tema, además de fomentar la difusión de la disciplina y la reafirmación de los derechos soberanos sobre nuestros recursos naturales. English: The goal of this work is to compile the existing information about the species present in our country. This has been done by means of datasheets that list both local references and reviews by foreign authors that include Argentine taxa. The main information sources consulted were Ringuelet et al. (1967), CLOFFSCA (2003), López et al. (2003; 2006), Liotta (2006) and the online databases of W. N. Eschmeyer and Fish Base. This is an open-ended publication. As such, it will require permanent updating, which will depend on the good will and collaboration of the ichthyological community. With this contribution, we hope to help to build a database of the freshwater fish of Argentina. The completion of this project will provide support to diverse plans emanating from different sectors, as well as promote the diffusion of this field of research and reaffirm Argentina’s sovereign rights over our natural resources. (Texto en English y Espanol. PDF tiene ocho paginas.)

2D Kinetics and Forced Dissociation of Selectin-ligand Bindings

Cell adhesion is crucial to many pathophysiological processes, such as inflammatory reaction and tumor metastasis. It is mediated by specific interactions between receptors and ligands, and provides the physical linkages among cells. For example, interactions between selectins and glycoconjugate ligands mediate leukocyte initially tethering to and subsequently rolling on vascular surfaces in sites of inflammation or injury, which is determined by their fast kinetic rates. To mediate cell adhesion, the interacting receptors and ligands must anchor to apposing surfaces of two cells or a cell and the substratum, i.e. , the so-called two-dimensional (2D) binding, which differs from interactions in the fluid phase, i.e. , the three-dimensional (3D) binding. How structural variations and surface environments of interacting molecules affect their 2D kinetics, and how external forces manipulate their dissociation has little been known quantitatively, and nowadays attracts more and more attentions.

An assessment of chemical contaminants in the marine sediments of southwest Puerto Rico

This report summarizes the results of a characterization of chemical contaminants in the sediments in southwest Puerto Rico. The report is part of a project to integrate various analytical specialties to assess linkages between chemical contaminants and the condition of coral reefs. In this phase of the project, over 120 chemical contaminants were analyzed in sediments collected, including a number of organic (e.g., hydrocarbons), inorganic (e.g., metals), and biological (bacterial) compounds/analytes. The report also provides a preliminary analysis of the association between sediment contaminants and coral species richness. Overall, the levels of chemical contaminants in the study area between Guanica Bay and the town of La Parguera were fairly low. At most of the sites sampled, particularly adjacent to the town of La Parguera, concentrations of organic and inorganic contaminants were below the median values from NOAA’s National Status and Trends Program, which monitors the Nation’s coastal and estuarine waters for chemical contaminants. Elevated levels of a number of contaminant classes were seen at the two sites sampled within Guanica Bay. An initial analysis of modeled PAH (hydrocarbon) data and coral species richness (reef building species) indicated a strong negative correlation between the presence of PAHs in the sediments and coral species richness. Additional work is needed to assess possible reasons for this observed pattern. (PDF contains 126 pages).

PICES Press, Vol. 16, No. 2, July 2008

The 2008 Inter-Sessional Science Board Meeting (pp.1-2, pdf, 0.1 Mb) FUTURE – From Science Plan to Implementation Plan (pp. 3-4, pdf, 0.1 Mb) CFAME Task Team Workshop – Linking and Visualising (p. 5, pdf, 0.1 Mb) PICES WG 21 Meets in Busan, Korea: The Database Meeting (pp. 6-7, pdf, 0.1 Mb) ICES-PICES-IOC Symposium on Climate Change (pp. 8-12, pdf, 1.2 Mb) Zooplankton and Climate: Response Modes and Linkages (pp. 13-15, pdf, 0.2 Mb) PICES Fishery Science Committee Workshop in Gijón (pp. 16-18, pdf, 0.1 Mb) The North Pacific Continuous Plankton Recorder Survey (pp. 19-21, pdf, 0.4 Mb) PICES Ecosystem Status Report Wins Design Award (p. 21, pdf, 0.4 Mb) Canada’s Three Oceans (C3O): A Canadian Contribution to the International Polar Year (pp. 22-25, pdf, 0.8 Mb) New Surface Mooring at Station Papa Monitors Climate (pp. 26-27, pdf, 0.2 Mb) The State of the Western North Pacific in the Second Half of 2007 (pp. 28-29, pdf, 0.4 Mb) The Bering Sea: Current Status and Recent Events (pp. 30-31, pdf, 0.4 Mb) Recent Trends in Waters of the Subarctic NE Pacific (pp.32-33, pdf, 0.3 Mb) 2009 Vintage of Fraser River Sockeye Salmon: A Complex Full Bodied Redd with Mysterious Bouquet (p. 34, pdf, 0.1 Mb) Pacific Biological Station Celebrates Centennial Anniversary, 1908–2008 (p. 35, pdf, 0.3 Mb) Marine and Coastal Fisheries: American Fisheries Society Open Access E-journal (p. 36, pdf, 0.1 Mb) Latest and Upcoming PICES Publications (p. 36, pdf, 0.1 Mb)