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The ability of Desulfovibrio vulgaris Hildenborough to reduce, and therefore contain, toxic and radioactive metal waste has made all factors that affect the physiology of this organism of great interest. Increased salinity is an important and frequent fluctuation faced by D. vulgaris in its natural habitat. In liquid culture, exposure to excess salt resulted in striking elongation of D. vulgaris cells. Using data from transcriptomics, proteomics, metabolite assays, phospholipid fatty acid profiling, and electron microscopy, we used a systems approach to explore the effects of excess NaCl on D. vulgaris . In this study we demonstrated that import of osmoprotectants, such as glycine betaine and ectoine, is the primary mechanism used by D. vulgaris to counter hyperionic stress. Several efflux systems were also highly up-regulated, as was the ATP synthesis pathway. Increases in the levels of both RNA and DNA helicases suggested that salt stress affected the stability of nucleic acid base pairing. An overall increase in the level of branched fatty acids indicated that there were changes in cell wall fluidity. The immediate response to salt stress included up-regulation of chemotaxis genes, although flagellar biosynthesis was down-regulated. Other down-regulated systems included lactate uptake permeases and ABC transport systems. The results of an extensive NaCl stress analysis were compared with microarray data from a KCl stress analysis, and unlike many other bacteria, D. vulgaris responded similarly to the two stresses. Integration of data from multiple methods allowed us to develop a conceptual model for the salt stress response in D. vulgaris that can be compared to those in other microorganisms.

Rising world demand for petroleum and natural gas makes the Caspian Sea and its resources the focus of increased international attention. This article examines the controversy over the legal condition of the Caspian Sea, both in terms of delimination questions and its status as a lake or a semi-enclosed sea. The natural resource potential of the Caspian is assessed, with a view to highlighting its hydrocarbon and fishery resources, as well as the environmental implications posed by development of industry and pipelines throughout the region. Also examined are the geopolitical priorities of the five Caspian littoral states, as well as the need to maintain the region as a zone for peaceful commercial enterprise and economic development. Toward these ends, the study concludes with an evaluation of the realistic prospects for collaboration among the Caspian states to protect the environment as evidenced by the recent negotiation of a special convention dedicated to that purpose.

Gas chromatography–multiplex coherent Raman (GC-MCR) is a new tandem technique that can be used for the high-speed analysis of volatile mixtures. BTEX serves as a useful and challenging test sample because of the similarity in boiling point and spectroscopic properties of its constituents. The ability to spectroscopically resolve isomers (e.g., m-xylene and p-xylene) allows GC-MCR to sacrifice chromatographic resolution for speed. The result is the analysis of BTEX in less than 5 min, which is relatively fast compared with other tandem GC techniques.

The ability to separate and sort peaks is explored using a new coherent two-dimensional form of resonance Raman spectroscopy. This experimental technique distributes normally congested rotational-vibrational peaks along a series of curved lines according to vibrational sequence, rotational quantum number, and selection rule. Each line consists of rotational-vibrational peaks that have the same vibrational sequence and the same value for DeltaJ, distributed in order by rotational quantum number. For diatomic molecules, these lines originate from points where they initially travel in opposite or orthogonal directions in two-dimensional space, which helps facilitate the separation between lines. Simulations and experimental results on C2 in a flame confirm the ability to separate and sort these normally congested rotational-vibrational peaks. This method appears to provide a solution to the long-standing problems of spectral congestion and disorder in gas-phase electronic spectra.

As the international community responds to the September 11 attacks and the general war on terrorism, member states in the United Nations (UN) must continue to address the balance between the rights of the individual and the security of the state. This study highlights the various roles that the UN plays in combating transnational terrorism through norm setting, codification of human rights law, as well as the drafting and effective implementation of 12 key multilateral agreements aimed at counter‐terrorism. In these ways, the UN provides direction and guidance for implementing human rights rules and the ways in which they should be enforced. Even so, national governments must make these rules work. In a system of sovereign states, the role of the UN organization in checking or reversing these human rights abuses remains severely limited and largely dependent upon the political will of the member states. As a consequence, part of the price paid for protecting national security against threats posed by terrorism may well be the curtailment of some human rights and civil liberties within the liberal democratic state.

A new detector for gas chromatography has been developed that is based upon nonlinear Raman spectroscopy. The resulting hyphenated instrument uses a simple windowless cell as the interface between the chromatograph and the spectroscopic detector. The eluents are detected using a unique spectrometer equipped with a broadband OPO. This spectrometer is capable of generating high-resolution coherent Raman signals that are suitable for rapid multichannel detection.

A new Raman spectrometer has been developed that can produce a complete high-resolution gas-phase vibrational spectrum during the time period of a single laser pulse. Conventional gas-phase Raman spectroscopy uses a single narrow-band laser to produce signals that are weak, requiring relatively long collection times. This spectrometer uses a degenerate OPO as a broad-band source that covers a continuous range of >3000 cm-1. When a beam from this broad-band source is added to a narrow-band laser beam, the intensity of the Raman signal is amplified by the nonlinear optical effect. The resulting instrument can generate spectra with high-spectral (

The Deepwater Horizon (DWH) oil spill in the spring of 2010 resulted in an input of ∼4.1 million barrels of oil to the Gulf of Mexico; >22% of this oil is unaccounted for, with unknown environmental consequences. Here we investigated the impact of oil deposition on microbial communities in surface sediments collected at 64 sites by targeted sequencing of 16S rRNA genes, shotgun metagenomic sequencing of 14 of these samples and mineralization experiments using 14 C-labeled model substrates. The 16S rRNA gene data indicated that the most heavily oil-impacted sediments were enriched in an uncultured Gammaproteobacterium and a Colwellia species, both of which were highly similar to sequences in the DWH deep-sea hydrocarbon plume. The primary drivers in structuring the microbial community were nitrogen and hydrocarbons. Annotation of unassembled metagenomic data revealed the most abundant hydrocarbon degradation pathway encoded genes involved in degrading aliphatic and simple aromatics via butane monooxygenase. The activity of key hydrocarbon degradation pathways by sediment microbes was confirmed by determining the mineralization of 14 C-labeled model substrates in the following order: propylene glycol, dodecane, toluene and phenanthrene. Further, analysis of metagenomic sequence data revealed an increase in abundance of genes involved in denitrification pathways in samples that exceeded the Environmental Protection Agency (EPA)'s benchmarks for polycyclic aromatic hydrocarbons (PAHs) compared with those that did not. Importantly, these data demonstrate that the indigenous sediment microbiota contributed an important ecosystem service for remediation of oil in the Gulf. However, PAHs were more recalcitrant to degradation, and their persistence could have deleterious impacts on the sediment ecosystem. © 2014 International Society for Microbial Ecology All rights reserved.

Worldwide interconnectivity through massive computer networks now makes states vulnerable to new threats. Foreign governments can launch computer-based assaults, or acts of information warfare, on another state's domestic systems such as energy grids, telecommunications, and financial facilities that could severely damage or disrupt national defence or vital social services. Even realizing the new forms of computer-generated weapons and changing concepts of sovereignty and territory brought about by global interdependence, international law is likely to rely on UN Charter principles to define the legal boundaries of cyberspace. While perhaps not armed force literally, resort to cyberforce may be viewed as a form of intervention that can produce harmful or coercive effects, and put at risk the national security of another state. There is need for modern international law to define more precisely the criteria used to distinguish which state actions are permissible as normal computer- generated transborder data flow from those cyberactivities that might qualify as an 'armed attack' against a state. Clearer rules are also needed for what responses are permissible as self-defence by a state targeted in an information warfare situation and how international institutions might facilitate the attainment of these objectives.

OBJECTIVESWe sought to evaluate whether oral anticoagulant (AC) therapy given for five months was superior to standard (control) therapy in patients with unstable angina receiving aspirin.BACKGROUNDThe long-term risk of myocardial infarction (MI) or death remains high in patients with unstable angina, despite the use of aspirin. Therefore, additional treatments are necessary.METHODSOf the 10,141 patients entering the main trial, 3,712 were randomized 12 to 48 h later to receive oral AC therapy (n = 1,848) or standard therapy (n = 1,864).RESULTSOne-hundred forty patients (7.6%) suffered from cardiovascular death, MI or stroke while receiving oral AC, compared with 155 patients (8.3%) on standard therapy (relative risk [RR] 0.90, 95% confidence interval [CI] 0.72 to 1.14; p = 0.40). The rates of the primary outcomes plus refractory angina were 16.7% (n = 308) versus 17.5% (n = 327) (RR 0.95, 95% CI 0.81 to 1.11; p = 0.53). Countries were divided into good or poor compliers (based on the use of oral AC above or below 70% at 35 days), without knowledge of results by country. In good-complier countries, oral AC was discontinued in only 10.4% of patients at seven days and in 23.6% by five months, compared with 27.6% and 44.9%, respectively, in poor complier countries. There were significant reductions in the risks of both the primary (6.1% vs. 8.9%; RR 0.68, 95% CI 0.48 to 0.95; p = 0.02) and secondary outcomes (11.9% vs. 16.5%; RR 0.70, 95% CI 0.55 to 0.90; p = 0.005) with oral AC in the good-complier countries. There was little difference in the poor-complier countries (9.0% vs. 7.8% for the primary and 21.3% vs. 18.5% for the secondary outcomes, tests for interactions comparing the RRs for the primary and secondary outcomes were p < 0.02 and p = 0.002, respectively, between the two sets of countries). In the overall study, there was an excess of major bleeding (2.7% vs. 1.3%; p = 0.004), which was larger in the good-complier countries (RR 2.71) compared with the poor-complier countries (RR 1.58). There were also reductions in cardiac catheterization (RR 0.80; p = 0.004) and coronary revascularization procedures (RR 0.82; p = 0.06) in the good-complier countries, but not in the poor-complier countries (RR 0.98 and 1.06, respectively, p for interaction of 0.06 and 0.04, respectively).CONCLUSIONSOverall, oral AC led to a small, nonsignificant reduction in the risk of the primary and secondary outcomes. Stratifying the countries or centers by their rates of compliance to oral AC suggested that good compliance to oral AC could potentially lead to clinically important reductions in major ischemic cardiovascular events.

Background Other than aspirin, there are few oral antithrombotic treatments with proven efficacy in patients with acute coronary syndrome. In this report, we present the rationale, design and baseline characteristics of the Clopidogrel in Unstable angina to prevent Recurrent ischaemic Events (CURE) trial, which includes a meta-analysis of the effects of thienopyridines in patients with vascular disease. Methods and Results Combined data from randomized trials of thienopyrindines in patients with atherosclerotic disease demonstrated a 29% reduction in vascular events when compared with placebo/control (n=2392) (OR 0.71, 95% CI 0.58-0.86, P=0.0006) and a 10% reduction in vascular events when compared with aspirin (n=22 254) (OR 0.91, 95% Cl 0.84-0.99, P=0.039). Similarly, randomized trials of aspirin plus thienopyridines in patients undergoing intracoronary stenting, demonstrated marked benefit of aspirin plus ticlopidine in reducing death or myocardial infarction compared with aspirin alone (OR 0.23, 95% CI 0.11-0.49, P=0.0001) or aspirin plus warfarin (OR 0.51, 95% CI 0.33-0.78, P=0.002). Whether these benefits extend to the much larger population of patients with acute coronary syndrome is unknown. CURE is an international, randomized, double-blind trial, in which patients with acute coronary syndrome will be randomized to receive either a bolus dose of clopidogrel (300 mg) followed by 75 mg per day for 3-12 months, or matching placebo. Both groups will receive aspirin. The co-primary efficacy end-points of CURE are: (1) the composite of cardiovascular death, myocardial infarction or stroke; and (2) the composite of cardiovascular death, myocardial infarction, stroke or refractory ischaemia. CURE will recruit approximately 12 500 patients with acute coronary syndrome (from 28 countries) and its power to detect moderate treatment benefits will be in the region of 80-90%, while maintaining an overall type I error (a) of 0.05. The baseline characteristics of the study population are consistent with at least a moderate risk group of patients with acute coronary syndrome. Conclusions Randomized trials of thienopyridines in patients with vascular disease demonstrate that thienopyridines are effective in reducing vascular events when compared with placebo/control or aspirin, as well as when used in combination with aspirin in patients undergoing intracoronary stent implantation. The CURE trial is a large international study to determine if acute and longterm treatment with the combination of clopidogrel and aspirin is superior to aspirin alone in patients with acute coronary syndrome. (C) 2000 The European Society of Cardiology.

This study surveys the international regime governing the use of ocean space by examining various agreements that have come to comprise it. While the regime for the oceans is commonly viewed within the context of a general, universal convention for the law of the sea, the contemporary scope of global oceans law extends far beyond that single instrument. Indeed, the law of the sea is broad, far-reaching and reflective of the increasingly complex, interdependent nature of contemporary international relations. In this regard, by end of the 20th century there has emerged a pronounced trend toward worldwide participation in and adherence to the multifaceted legal regime established to manage the oceans. This panorama of legal instruments focuses both on international instruments and regional accords. The study's premise is this: Since 1958 several multilateral agreements have been negotiated and implemented specifically to regulate various uses of ocean space. Viewed as an integrated whole, these agreements comprise at the beginning of the 21st century an intertwined, sophisticated international legal regime for managing ocean activities. The range of agreements discussed here, however, is neither exhaustive nor meant to suggest that sufficient law is available for regulating all activities affecting the oceans now or in the future. Rather, the survey furnishes a starting point for appreciating just how far the international community has come in establishing a legal regime for managing people and their technological activities that impact on ocean space.

During the 1990s, the World Trade Organization (WTO) rejected as impermissible unilateral efforts by the United States to promote conservation of endangered marine species through trade sanctions against other governments. These acts of unilateral economic coercion were held to be discriminatory tactics done in unfair restraint of international trade. But in so doing, the WTO findings aroused the ire of environmentalists worldwide. These findings by the WTO became portrayed not as decisions upholding free trade, but as mandates against marine conservation and environmental protection. Even so, this denial of lawful permissibility to use unilateral economic coercion to protect endangered species internationally does not signal the demise of national efforts to conserve living marine resources. Rather, these WTO findings point up the manner in which potential trade and marine conservation disputes should be handled, i.e., through means of peaceful settlement. The key to future international marine conservat...

he UN has been maligned over the past decade for peacekeeping failures in Somalia and Bosnia and for being a bloated, inefficient, JL ineffective bureaucracy. Yet, at the same time, the organization has become increasingly active in promoting the democratic process around the world, particularly in developing countries. It is, in fact, not hyperbole to assert that the UN Secretariat has assumed the impressive role of inter national agent for democratization, as it has engaged the UN organization in various activities toward that end in more than seventy-five states and territories over the past decade. Unfortunately, this formidable UN effort at assisting the democratic process worldwide has gone largely neglected and unappreciated. This study aims to remedy this neglect by pursuing three main pur poses. First, I examine the concept of "democracy" as interpreted and ap plied by the UN so that a clearer picture can be derived of the particular political system envisioned, promoted, and supported by the UN Secre tariat. I also offer an appraisal of the Secretariat's rationale for undertak ing such efforts at promoting democracy and the special role, if any, that certain member states might play in these efforts. Second, I examine the ways and means the UN has adopted for im plementing assistance to states that have requested special help from the UN in the democratization process. This analysis provides insights into how the UN facilitates the democratic process and which states have been most active in seeking UN support. For the discussion, a clearer picture emerges of which facets of democracy are considered paramount by the UN and how the organization has contributed to institutionalizing those facets in various countries around the world. A third purpose of the study is to gauge what success the UN has had in its efforts to promote democracy. I do this by examining certain key questions, among them: Have the UN's efforts been worthwhile? Where has the UN succeeded in promoting democracy, and where has it failed? Are there political, legal, and ideological lessons to be learned from the UN experience in promoting democracy? In sum, can the UN make a dif ference in promoting the acceptability of democracy worldwide? If so, how, and to what degree? Finally, the conclusion aims to draw lessons

Part I. The UN System as a Source of Law: 1. The UN legal order: an overview Oscar Schachter 2. General law-making processes Paul Szasz 3. Specialized law-making processes Frederic L. Kirgis, Jr Part II. Substantive Law: 4. Force and arms John F. Murphy 5. Human rights Hurst Hannum 6. Refugees and migration David A. Martin 7. Women Rebecca J. Cook 8. Labor Virginia A. Leary 9. Economic relations and development Stephen Zamora 10. Environment Ved P. Nanda 11. Law of the sea Bernard H. Oxman 12. Outer space Ralph G. Steinhardt 13. International crimes John F. Murphy Part III. International Law: 14. Law of the international civil service Robert S. Jordan 15. Financial responsibility of members Jose E. Alvarez 16. Conclusion: the United Nations as international law-giver Christopher C. Joyner.

The United States has important national interests vested in both the Arctic and Antarctic Oceans. Thus, in recent decades the United States has progressively codified its national commitment to conserve and manage both these marine regions and their resources and to protect them from activities that might produce adverse impacts. The U.S. legislation passed since the 1960s selectively affects both polar regions and supports major American policy objectives there: to maintain the Arctic and Antarctic as areas of international cooperation for peaceful purposes; to satisfy economic needs, especially hydrocarbon, mineral, and living resources; to protect opportunities for scientific research; to protect the marine environment; and to conserve living resources in the circumpolar seas. Future trends suggest a shift in U.S. policy attention to the Arctic. Accordingly, new U.S. legislation will be needed to regulate increased activities in the polar north.

The Spratly Islands archipelago has become pivotal as a strategic, economic and political asset in the South China Sea. This situation has become particularly evident since the end of the Cold War, as the Spratlys have been used by six littoral states as legal basepoints to project claims over strategic sea-lanes, fishery waters and submarine hydrocarbon resources in the South China Sea. The claimant state having predominant military and economic influence in the region is China, which has used military force on occasion to substantiate its claims. It is not inconceivable that the dispute over the Spratlys could lead to military conflict. Successful settlement of sovereignty disputes for the Timor Gap area, Antarctica and the Svalbard archipelago provide models for creating a joint resource development authority to promote regional co-operation among Spratly claimants. The most critical ingredient for mitigating tensions in the South China Sea is the political willingness of Spratly claimants, especially China, to make dispute settlement happen.

The N- and C-terminal domains of human somatic angiotensin I converting enzyme (sACE-1) demonstrate distinct physiological functions, with resulting interest in the development of domain-selective inhibitors for specific therapeutic applications. Herein, the activity of lisinopril-coupled transition metal chelates was tested for both reversible binding and irreversible catalytic inactivation of each domain of sACE-1. C/N domain binding selectivity ratios ranged from 1 to 350, while rates of irreversible catalytic inactivation of the N- and C-domains were found to be significantly greater for the N-domain, suggesting a more optimal orientation of M-chelate-lisinopril complexes within the active site of the N-domain of sACE-1. Finally, the combined effect of binding selectivity and inactivation selectivity was assessed for each catalyst (double-filter selectivity factors), and several catalysts were found to cause domain-selective catalytic inactivation. The results of this study demonstrate the ability to optimize the target selectivity of catalytic metallopeptides through both binding and catalytic factors (double-filter effect).

Most drugs function by binding reversibly to specific biological targets, and therapeutic effects generally require saturation of these targets. One means of decreasing required drug concentrations is incorporation of reactive metal centers that elicit irreversible modification of targets. A common approach has been the design of artificial proteases/nucleases containing metal centers capable of hydrolyzing targeted proteins or nucleic acids. However, these hydrolytic catalysts typically provide relatively low rate constants for target inactivation. Recently, various catalysts were synthesized that use oxidative mechanisms to selectively cleave/inactivate therapeutic targets, including HIV RRE RNA or angiotensin converting enzyme (ACE). These oxidative mechanisms, which typically involve reactive oxygen species (ROS), provide access to comparatively high rate constants for target inactivation. Target-binding affinity, co-reactant selectivity, reduction potential, coordination unsaturation, ROS products (metal-associated vs metal-dissociated; hydroxyl vs superoxide), and multiple-turnover redox chemistry were studied for each catalyst, and these parameters were related to the efficiency, selectivity, and mechanism(s) of inactivation/cleavage of the corresponding target for each catalyst. Important factors for future oxidative catalyst development are 1) positioning of catalyst reduction potential and redox reactivity to match the physiological environment of use, 2) maintenance of catalyst stability by use of chelates with either high denticity or other means of stabilization, such as the square planar geometric stabilization of Ni- and Cu-ATCUN complexes, 3) optimal rate of inactivation of targets relative to the rate of generation of diffusible ROS, 4) targeting and linker domains that afford better control of catalyst orientation, and 5) general bio-availability and drug delivery requirements.

The 1991 Protocol on Environmental Protection to the Antarctic Treaty and its annexes contribute substantially to contemporary international law prohibiting marine pollution. As the Protocol approaches obtaining the number of ratifications necessary for entry into force, its relevance for international ocean law accordingly becomes more salient. Thus, measures in the Protocol and its annexes take on particular importance, as they are made to apply to all sources of pollution affecting the Southern Ocean ecosystem (i.e., sources from land, dumping, vessels, the atmosphere and the seabed). Such an all-encompassing approach well serves to bolster both the normative and policy framework for effecting anti-pollution marine law in the region.

The Third United Nations Conference on the Law of the Sea (UNCLOS III) provides valuable lessons for future formulation of law to govern global commons, areas that lie beyond the limits of national jurisdiction and to which all peoples have free and open access. Although endowed with the advantage of a certain degree of scientific certainty about the need for regulation of the ocean environment, UNCLOS III fell victim to a North‐South schism that impaired the search for consensus on important issues and undermined the final product of the negotiations. An examination of the UNCLOS III experience suggests that agreements that exclude specially affected states are unlikely to succeed, and points to a variety of advantages and disadvantages that come from linking several issues under one negotiating framework. The consensus approach to negotiation used at the Conference tends to expand the time and effort needed to reach a successful outcome, which can lead the negotiations themselves to be outstripped by te...

Canada's seizure of the Spanish fishing vessel Estai in March 1995 touched off severe political tensions between Canada and the European Community. Known as the "Turbot War", the dispute arose over quota levels agreed to by the Northwest Atlantic Fisheries Organization for turbot stocks in fishing grounds straddling the high seas beyond Canada's proclaimed 200-mile national fishery zone. While underscoring tensions between coastal states and foreign fishing nations, this dispute became a diplomatic catalyst for the promulgation in August 1995 of a special UN Convention on the Conservation and Management of Straddling Fish Stocks and Highly Migratory Stocks. The "Turbot War" thus highlighted the utility of regional fishery organizations in monitoring and enforcing international fishing regulations and strengthened the international regulatory framework for managing straddling stocks under the 1982 LOS Convention.

This article examines the objections by the United States to the 1982 Law of the Sea Convention and explains why the government rejected a treaty that it had worked so long and hard to produce. This study also evaluates UN efforts since 1990 to reconcile international difficulties over seabed mining. The focus here falls on how deep seabed provisions of the 1982 text were revised to render the Convention more acceptable to the United States. Certain lessons for multilateral negotiations from the protracted law of the sea experience are also proffered, especially how circumstances of time, economics, politics, and personalities can affect the outcomes of complex international negotiations. Finally, the 1994 Implementation Agreement is assessed. The conclusion is that this Agreement makes notable improvements benefitting U.S. interests as a principal ocean miner. It replaces an overly detailed and costly regime in Part XI of the 1982 Law of the Sea Convention text with a more streamlined approach to deep se...

Forging Peace: Intervention, Human Rights and the Management of Media Space. Edited by Monroe E. Price, Mark Thompson. Bloomington: Indiana University Press, 2002. 416 pp., $64.95 cloth (ISBN: 0-253-34197-3), $27.95 paper (ISBN: 0-253-21573-0). How should the international community react to the incendiary use of the media to encourage genocide during civil wars, as occurred in Rwanda and Bosnia? How can international peace operations confront and overcome the manipulative propaganda often used by host governments to incite violence against segments of their populations? What roles do international legal rules play in forging acceptable standards and practices for the media in war-torn societies? These questions underpin the main aim of Forging Peace , namely, to examine the ways in which international organizations, both governmental and nongovernmental, have promoted pluralist, independent media in recent postconflict situations. The actions advocated—called information intervention—offer feasible and lawful ways to prevent mass human rights atrocities, the leitmotiv of this volume. Forging Peace begins with an excellent introductory chapter by Mark Thompson and Monroe Price that spells out the characteristics and instruments of information intervention as a lawful instrument of human rights protection. Thereafter, the volume is divided into four sections. The first section contains an interview by Mark Thompson of Jamie Metzl, in which Metzl elaborates on his original definition of information intervention (Metzl 1997). Having thus defined the concept, the second section analyzes the moral, legal, and practical implications of information intervention. In an authoritative discussion, Stephanie Farrior assesses hate propaganda's relevance for human rights law. Such broadcasts violate the norms of decency inherent in free information freely broadcast, and Farrior rightly concludes that we need to rethink the legal status of such pernicious misinformation. In a companion piece, Eric Blinderman critiques the lawfulness of information intervention as it relates to sovereignty under international legal rules. Although current law prohibits jamming broadcasts within a state …

Artificial nucleases containing Rev-coupled metal chelates based on combinations of the transition metals Fe(2+), Co(2+), Ni(2+), and Cu(2+) and the chelators DOTA, DTPA, EDTA, NTA, tripeptide GGH, and tetrapeptide KGHK have been tested for DNA nuclease activity. Originally designed to target reactive transition metal chelates (M-chelates) to the HIV-1 Rev response element mRNA, attachment to the arginine-rich Rev peptide also increases DNA-binding affinity for the attached M-chelates. Apparent K(D) values ranging from 1.7 to 3.6 µM base pairs for binding of supercoiled pUC19 plasmid DNA by Ni-chelate-Rev complexes were observed, as a result of electrostatic attraction between the positively-charged Rev peptide and negatively-charged DNA. Attachment of M-chelates to the Rev peptide resulted in enhancements of DNA nuclease activity ranging from 1-fold (no enhancement) to at least 13-fold (for Cu-DTPA-Rev), for the rate of DNA nicking, with second order rate constants for conversion of DNA(supercoiled) to DNA(nicked) up to 6 × 10(6) M(-1) min(-1), and for conversion of DNA(nicked) to DNA(linear) up to 1 × 10(5) M(-1) min(-1). Freifelder-Trumbo analysis and the ratios of linearization and nicking rate constants (k(lin)/k(nick)) revealed concerted mechanisms for nicking and subsequent linearization of plasmid DNA for all of the Rev-coupled M-chelates, consistent with higher DNA residency times for the Rev-coupled M-chelates. Observed rates for Rev-coupled M-chelates were less skewed by differing DNA-binding affinities than for M-chelates lacking Rev, as a result of the narrow range of DNA-binding affinities observed, and therefore relationships between DNA nuclease activity and other catalyst properties, such as coordination unsaturation, the ability to consume ascorbic acid and generate diffusible radicals, and the identity of the metal center, are now clearly illustrated in light of the similar DNA-binding affinities of all M-chelate-Rev complexes. This work paints a clearer picture of the factors governing DNA nuclease activity by redox active M-chelates than was previously possible. The results demonstrate enhancement of DNA cleavage by use of a targeting sequence, but also clearly underscore that significant orientational factors are required for optimal reactivity at the metal center. Moreover, the studies confirm high selectivity for the target HIV RRE RNA at the most likely dosage concentrations, lending further support to the feasibility of designing and applying targeted catalytic metallodrugs.

Research indicates that chronic daily cocaine injections produce sensitization to, while the chronic continuous infusion of cocaine produces tolerance to, its behavioral and neurochemical effects. The present experiments examined whether flupenthixol administration during withdrawal would attenuate/eliminate the behavioral effects produced by these administration regimens. The rats were pretreated for 14 days with either continuous or intermittent daily injections of cocaine, and were then withdrawn from the pretreatment regimen for 7 days. On days 1-5 of the withdrawal period, subjects received a daily 0.125-2.0 mg/kg IP injection of flupenthixol. Then on day 7 of withdrawal from the cocaine pretreatment, all rats were given a 15.0 mg/kg IP injection of cocaine. Their behavior was rated according to a modified version of the Ellinwood and Balster (6) scale for 60 min. The results indicated that flupenthixol treatment during withdrawal eliminated the tolerance normally associated with the continuous infusion of cocaine. However, this effect of flupenthixol was not dose dependent: the lowest dose had the same effect as the highest dose of flupenthixol. In the cocaine-injection subjects, flupenthixol had a slight but statistically significant reduction in the behavioral effects of cocaine. The same was true in the saline-control rats, except for the highest dose of flupenthixol, which had a significant enhancing effect on the behavioral response to cocaine. The present results suggest that the current procedures may represent an effective screening methodology for potential cocaine pharmacotherapies.

Research indicates that daily cocaine injections produce sensitization to, while the continuous infusion of cocaine produces tolerance to, its behavioral and neurochemical effects. The effects of the continuous infusion of cocaine are consistent with the withdrawal syndrome reported by human cocaine abusers. The present experiment examined whether amantadine administrations during withdrawal from continuous or intermittent cocaine attenuate and/or eliminate the behavioral effects produced by these administration regimens. The rats were pretreated for 14 days with either continuous or intermittent daily injections of cocaine, and were then withdrawn from the pretreatment regimen for 7 days. On days 1-5 of the withdrawal period, half the subjects received a 5.0 mg/kg IP injection of amantadine, and the other half received a 20.0 mg/kg IP injection of amantadine. On day 7 of withdrawal from the cocaine pretreatment, all rats were given a 15.0 mg/kg IP injection of cocaine. Their behavior was rated according to the modified Ellinwood and Balster (6) scale for 60 min. The results indicated that amantadine treatment during withdrawal eliminated the tolerance normally associated with the continuous infusion of cocaine. In contrast, in both the saline control and daily injection subjects amantadine treatment during withdrawal resulted in a slight, but statistically significant, reduction in the behavioral effects of cocaine. The present results therefore indicate that low doses of amantadine should be considered as a potential pharmacotherapy for the early stages of cocaine withdrawal. Furthermore, the present experimental procedures may represent an effective screening methodology for potential cocaine pharmacotherapies.

The present experiments examined alterations in 5-HT3 receptors during withdrawal from continuous or intermittent cocaine. Rats were pretreated with 40 mg/kg/day cocaine for 14 days by either SC injections or osmotic minipumps. The rats were then withdrawn from the pretreatment regimen for 7 days. In Experiment 1, rats received 0-16 mg/kg IP injections of ondansetron, a selective 5-HT3 receptor antagonist. In Experiment 2, the rats received 0-16 mg/kg IP ondansetron in combination with a 15 mg/kg IP injection of cocaine. In Experiment 3, the subjects received 0-16 mg/kg IP injections of ondansetron in combination with a 7.5 mg/kg IP injection of cocaine. Following these injections, the subjects' behavior was rated using the Ellinwood and Balster (18) rating scale. The results of Experiment 1 indicated that ondansetron had no effect on the behavior of the subjects, nor was there a differential effect of pretreatment regimen the effects of ondansetron. The results of Experiment 2 indicated that ondansetron had no effect on cocaine-induced locomotion in the saline control rats, but did have a slight, statistically significant, suppressive effect in the injection rats. In contrast, ondansetron had a robust facilitative effect on cocaine-induced locomotion in the continuous infusion rats. The results of Experiment 3 indicated that ondansetron had no effect on cocaine-induced locomotion in the saline control rats or the cocaine injection pretreatment subjects. In the continuous infusion subjects, ondansetron did have a slight, statistically significant, facilitative effect on cocaine-induced locomotion.(ABSTRACT TRUNCATED AT 250 WORDS)

Protein–protein interactions offer an insight into cellular processes beyond what may be obtained by the quantitative functional genomics tools of proteomics and transcriptomics. The aforementioned tools have been extensively applied to study Escherichia coli and other aerobes and more recently to study the stress response behavior of Desulfovibrio vulgaris Hildenborough, a model obligate anaerobe and sulfate reducer and the subject of this study. Here we carried out affinity purification followed by mass spectrometry to reconstruct an interaction network among 12 chromosomally encoded bait and 90 prey proteins based on 134 bait-prey interactions identified to be of high confidence. Protein-protein interaction data are often plagued by the lack of adequate controls and replication analyses necessary to assess confidence in the results, including identification of potential false positives. We addressed these issues through the use of biological replication, exponentially modified protein abundance indices, results from an experimental negative control, and a statistical test to assign confidence to each putative interacting pair applicable to small interaction data studies. We discuss the biological significance of metabolic features of D. vulgaris revealed by these protein-protein interaction data and the observed protein modifications. These include the distinct role of the putative carbon monoxide-induced hydrogenase, unique electron transfer routes associated with different oxidoreductases, and the possible role of methylation in regulating sulfate reduction.

Despite rapid evolution in international fisheries law and establishment of the exclusive economic zone (EEZ), straddling stocks still remain susceptible to heavy harvesting in high seas areas by distant‐water fishing states there by undermining coastal state management. The notion mar presencial (presential sea) has recently been proposed by Chile as a solution for the problem of straddling stocks. The presential sea concept was nationally designed and promoted to curtail such foreign fishing in areas adjacent to Chile's EEZ. This article examines the presential sea as a geostrategic concept, its justification for being, and the question of its permissibility under contemporary international fisheries law. Attention is also given to recent international developments that challenge the legal viability of the presential sea concept. The authors conclude that if this concept were to be widely adopted by coastal states, the traditional freedom to fish on the high seas might be severely compromised. ...

Microbes live in changing environments and change their phenotype via gene regulation in response. Although this transcriptional response is important for fitness, very little is known about how it evolves in microbes. We started by asking a number of high-level questions about the evolution of transcriptional phenotype: (1) To what extent is transcriptional response conserved, i.e. do conserved genes respond similarly to the same condition; (2) To what extent are transcriptional modules conserved; and (3) Does there exist a general stress response to a variety of stressors? To illuminate these questions, we analyzed more than 500 microarray experiments across the bacterial domain. We looked for conservation of transcriptional regulation both in close sister species and vastly divergent clades. In addition, we produced and analyzed an extensive in-house compendium of environmental stress data in three metal-reducing bacteria.

Hexavalent Chromium is a widespread contaminant found in soil, sediment, and groundwater. In order to stimulate microbially-mediated reduction of Cr(VI), a poly-lactate compound (HRC) was injected into the Chromium-contaminated aquifer at the Hanford (WA) 100H site in 2004. Cr(VI) concentrations rapidly declined to below the detection limit and remained so for more than three years after injection. Based on the results of the bacterial community composition using high-density DNA 16S rRNA gene microarrays, we observed the community to transition through denitrifying, ironreducing and sulfate-reducing populations. As a result, we specifically focused isolation efforts on three bacterial species that were significant components of the community. Positive enrichments in defined anaerobic media resulted in the isolation of an iron-reducing Geobacter metallireducens-like isolate, a sulfate-reducing Desulfovibrio vukgaris-like strain and a nitrate-reducing Pseudomonas stutzeri-like isolate among several others. All of these isolates were capable of reducing Cr(VI) anoxically and have been submitted for genome sequencing to JGI. To further characterize the microbial, and geochemical mechanisms associated with in situ Cr(VI) reduction at the site, additional HRC was injected in 2008. The goal was to restimulate the indigenous microbial community and to regenerate the reducing conditions necessary for continued Cr(VI) bio-immobilization in the groundwater. Analysismore » of the microbial populations post-injection revealed that they recovered to a similar density as after the first injection in 2004. In this study, we present the results from our investigation into microbially-mediated Cr(VI) reduction at Hanford, and a comparison of the microbial community development following two HRC injections four years apart.« less

Sulfate-reducing bacteria have been extensively studied for their potential in heavy-metal bioremediation. However, the occurrence of elevated nitrate in contaminated environments has been shown to inhibit sulfate reduction activity. Although the inhibition has been suggested to result from the competition with nitrate-reducing bacteria, the possibility of direct inhibition of sulfate reducers by elevated nitrate needs to be explored. Using Desulfovibrio vulgaris as a model sulfate-reducing bacterium, functional genomics analysis reveals that osmotic stress contributed to growth inhibition by nitrate as shown by the upregulation of the glycine/betaine transporter genes and the relief of nitrate inhibition by osmoprotectants. The observation that significant growth inhibition was effected by 70 mM NaNO(3) but not by 70 mM NaCl suggests the presence of inhibitory mechanisms in addition to osmotic stress. The differential expression of genes characteristic of nitrite stress responses, such as the hybrid cluster protein gene, under nitrate stress condition further indicates that nitrate stress response by D. vulgaris was linked to components of both osmotic and nitrite stress responses. The involvement of the oxidative stress response pathway, however, might be the result of a more general stress response. Given the low similarities between the response profiles to nitrate and other stresses, less-defined stress response pathways could also be important in nitrate stress, which might involve the shift in energy metabolism. The involvement of nitrite stress response upon exposure to nitrate may provide detoxification mechanisms for nitrite, which is inhibitory to sulfate-reducing bacteria, produced by microbial nitrate reduction as a metabolic intermediate and may enhance the survival of sulfate-reducing bacteria in environments with elevated nitrate level.

Rats were pretreated with 40 mg/kg/day cocaine for 14 days by either subcutaneous injections or osmotic minipumps. Rats were then withdrawn from the pretreatment regime for 1 or 7 days and given a 20-mg/kg IP cocaine challenge (day 1) or a 0-, 20-, 40-, or 40-mg/kg IP cocaine challenge (day 7). The results indicate that rats receiving intermittent, daily injections exhibited sensitization to the behavioral effects of a cocaine challenge on days 1 and 7 of withdrawal. In contrast, rats receiving continuous cocaine exhibited tolerance to the behavioral effects of a cocaine challenge on days 1 and 7 of withdrawal. The present results support and extend previous research that indicates that the route and temporal pattern of administration influences the effects of chronic cocaine. Furthermore, the present results indicate that the continuous infusion paradigm may represent an alternative animal model of some aspects of high-dose cocaine abuse, as compared to the typical procedure of single, or multiple, daily cocaine injections.