The Operation of Gliders Under the International Law of the Sea

In contrast to traditional ocean observation systems such as research vessels and moorings, gliders, a specific category of autonomous underwater vehicles, can be operated individually or as fleets. These devices are designed to allow for continuously and remotely commanded ocean observation from the surface down to at least 1 km depth and deliver data with high horizontal and temporal resolution. This article assesses the legal framework and identifies the legal conditions that ought to be fulfilled in order to deploy and operate gliders in conformity with the international law of the sea.     

Russia’s Annexation of Crimea and the Passage of Ships Through Kerch Strait: A Law of the Sea Perspective

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This article discusses potential Ukrainian rights of passage through Kerch Strait against the background of restrictions on passage imposed by Russia since the annexation of Crimea, taking into account the ongoing arbitral proceedings under Annex VII of the UN Convention on the Law of the Sea. The article addresses first the question of the status of Kerch Strait and the Sea of Azov under the international law of the sea. In a second step, the article deals with the issue of potential Ukrainian passage rights through Kerch Strait based on the legal status discussed in the first section as well as relevant bilateral treaties between Ukraine and Russia.     

On the relationship between the number of negative slope regions in the voltage-current curve of the Hodgkin-Huxley model and its parameter values

 We consider the problem of the existence of a negative slope region (NSR) in the voltage-current curve of the neuronal membrane and the relationship between this phenomenon and the membrane parameters. For the Hodgkin-Huxley model it is proposed to determine the dependence of the number of NSR on the values of the maximal sodium (gˉ_Na) and potassium (gˉ_K) conductances. The method is suggested for constructing the boundaries on the (gˉ_Na, gˉ_K) plane, in passing through which the number of NSR changes to 1. Using the method we partition the (gˉ_Na, gˉ_K) plane into the regions corresponding to the curves with the different number of NSR. This number can be changed from 0 to 2 in changing the values of gˉ_Na and gˉ_K over the physiologically possible range. Received: 9 December 1993/Accepted in revised form: 10 February 1995     

The Cytological Mechanism of Low Fertility in the Naked Seed Rice

The low fertility of naked seed rice (NSR) was investigated by the following observations: somatic chromosome constitute, behavior of pollen mother cells (PMCs), the germination of mature pollen grains, the development of male and female gametes and the structure of the anther opening. The results indicated that somatic chromosomal number was 2n = 24, behavior of PMCs were normal and most of pollen grains could regularly develop further to mature male gametophytes in NSR. And dehiscence chamber and thickened endothecium cell (TEC) in numerous anthers of the NSR were developed abnormally after dicaryotic phase, result in few anthers complete opening and most partly opening or failure to opening, therefore much fewer of pollen grains attach on the stigma as compared with normal variety. Furthermore most of embryo sacs possessed abnormal structure and were sterile. All of above illustrated that the failure of the anther opening and the abortion of female gametophyte were main factors controlling the low seed-setting rate of the NSR.     

Coastal States in the South China Sea and Submissions on the Outer Limits of the Continental Shelf

The purpose of this article is to examine the outer continental shelf submissions made by the coastal states of the South China Sea and their potential impact on legal and political developments in the South China Sea. In accordance with the United Nations Convention on the Law of the Sea of 1982 and the guidelines of the Commission on the Limits of the Continental Shelf, coastal states are to establish the outer limits of their continental shelf where it extends beyond 200 nautical miles. Meeting this obligation is complicated in enclosed or semienclosed seas where there are maritime disputes such as in the South China Sea.     

Nitrogen metabolism in Sinorhizobium meliloti-alfalfa symbiosis: dissecting the role of GlnD and PII proteins

To contribute nitrogen for plant growth and establish an effective symbiosis with alfalfa, Sinorhizobium meliloti Rm1021 needs normal operation of the GlnD protein, a bifunctional uridylyltransferase/uridylyl-cleavage enzyme that measures cellular nitrogen status and initiates a nitrogen stress response (NSR). However, the only two known targets of GlnD modification in Rm1021, the PII proteins GlnB and GlnK, are not necessary for effectiveness. We introduced a Tyr→Phe variant of GlnB, which cannot be uridylylated, into a glnBglnK background to approximate the expected state in a glnD-sm2 mutant, and this strain was effective. These results suggested that unmodified PII does not inhibit effectiveness. We also generated a glnBglnK-glnD triple mutant and used this and other mutants to dissect the role of these proteins in regulating the free-living NSR and nitrogen metabolism in symbiosis. The glnD-sm2 mutation was dominant to the glnBglnK mutations in symbiosis but recessive in some free-living phenotypes. The data show that the GlnD protein has a role in free-living growth and in symbiotic nitrogen exchange that does not depend on the PII proteins, suggesting that S. meliloti GlnD can communicate with the cell by alternate mechanisms.     

Multiple regions of NSR1 are sufficient for accumulation of a fusion protein within the nucleolus

NSR1, a 67-kD nucleolar protein, was originally identified in our laboratory as a nuclear localization signal binding protein, and has subsequently been found to be involved in ribosome biogenesis. NSR1 has three regions: an acidic/serine-rich NH2 terminus, two RNA recognition motifs, and a glycine/arginine-rich COOH terminus. In this study we show that NSR1 itself has a bipartite nuclear localization sequence. Deletion of either basic amino acid stretch results in the mislocation of NSR1 to the cytoplasm. We further demonstrate that either of two regions, the NH2 terminus or both RNA recognition motifs, are sufficient to localize a bacterial protein, beta-galactosidase, to the nucleolus. Intensive deletion analysis has further defined a specific acidic/serine-rich region within the NH2 terminus as necessary for nucleolar accumulation rather than nucleolar targeting. In addition, deletion of either RNA recognition motif or point mutations in one of the RNP consensus octamers results in the mislocalization of a fusion protein within the nucleus. Although the glycine/arginine-rich region in the COOH terminus is not sufficient to bring beta-galactosidase to the nucleolus, our studies show that this domain is necessary for nucleolar accumulation when an RNP consensus octamer in one of the RNA recognition motifs is mutated. Our findings are consistent with the notion that nucleolar localization is a result of the binding interactions of various domains of NSR1 within the nucleolus rather than the presence of a specific nucleolar targeting signal.     

The U-Shaped Line and a Categorization of the Ocean Disputes in the South China Sea

A principal aspect of the territorial and boundary delimitation disputes in the South China Sea is the so-called U-shaped line. This article addresses the genesis and substantiation of the U-shaped line claims as well as the possible change in positions of the governments of the Republic of China and the People's Republic of China with respect to the historical waters claim, which is an integral part of their U-shaped line positions. A legal analysis of the various communications of the South China Sea players with respect to the U-shaped line helps to clarify and identify the nature of four kinds of legal disputes. It is also possible to differentiate the various degrees of difficulty involved in settling each of these disputes.     

NSR1 is required for pre-rRNA processing and for the proper maintenance of steady-state levels of ribosomal subunits.

NSR1 is a yeast nuclear localization sequence-binding protein showing striking similarity in its domain structure to nucleolin. Cells lacking NSR1 are viable but have a severe growth defect. We show here that NSR1, like nucleolin, is involved in ribosome biogenesis. The nsr1 mutant is deficient in pre-rRNA processing such that the initial 35S pre-rRNA processing is blocked and 20S pre-rRNA is nearly absent. The reduced amount of 20S pre-rRNA leads to a shortage of 18S rRNA and is reflected in a change in the distribution of 60S and 40S ribosomal subunits; there is no free pool of 40S subunits, and the free pool of 60S subunits is greatly increased in size. The lack of free 40S subunits or the improper assembly of these subunits causes the nsr1 mutant to show sensitivity to the antibiotic paromomycin, which affects protein translation, at concentrations that do not affect the growth of the wild-type strain. Our data support the idea that NSR1 is involved in the proper assembly of pre-rRNA particles, possibly by bringing rRNA and ribosomal proteins together by virtue of its nuclear localization sequence-binding domain and multiple RNA recognition motifs. Alternatively, NSR1 may also act to regulate the nuclear entry of ribosomal proteins required for proper assembly of pre-rRNA particles.     

Small-molecule inhibitors of nisin resistance protein NSR from the human pathogen Streptococcus agalactiae

Lantibiotics are antimicrobial peptides produced by Gram-positive bacteria and active in the nanomolar range. Nisin is the most intensely studied and used lantibiotic, with applications as food preservative and recognized potential for clinical usage. However, different bacteria that are pathogenic for humans and do not produce nisin, including Streptococcus agalactiae, show an innate resistance that has been related to the nisin resistance protein (NSR), a membrane-associated protease. Here, we report the first-in-class small-molecule inhibitors of SaNSR identified by virtual screening based on a previously derived structural model of the nisin/NSR complex. The inhibitors belong to three different chemotypes, of which the halogenated phenyl-urea derivative NPG9 is the most potent one. Co-administration of NPG9 with nisin yields increased potency compared to nisin alone in SaNSR-expressing bacteria. The binding mode of NPG9, predicted with molecular docking and validated by extensive molecular dynamics simulations, confirms a structure-activity relationship derived from the in vivo data. Saturation transfer difference-NMR experiments demonstrate direct binding of NPG9 to SaNSR and agree with the predicted binding mode. Our results demonstrate the potential to overcome SaNSR-related lantibiotic resistance by small molecules.     

Enhancement of inward current by serotonin in neurons of Aplysia

In RB cells of Aplysia, serotonin, in the presence of TEA, 4AP and Ba, elicits a voltage-dependent inward current. In Ba-TEA-4AP seawater, RB cells showed a negative slope region (NSR) in their current-voltage (I-V) relationship when measured at the end of 2-s commands from a holding potential of -60 mV. Addition of serotonin to the bathing solution enhanced the NSR. When holding potential was lowered to -10 mV, the NSR as well as the effects of serotonin were greatly reduced. Addition of 20 mM cobalt to the bathing solution blocked both the NSR and the inward current produced by serotonin. Changes in potassium concentration produced no consistent shift in voltage sensitivity nor change in amplitude of the current elicited by serotonin. Intracellular injection of cesium sufficient to broaden action potentials did not block the enhancement of NSR by serotonin. These results support the conclusion that in RB cells, serotonin produces a voltage-dependent current carried by calcium ions.     

Role for nucleolin/Nsr1 in the cellular localization of topoisomerase I

Nucleolin functions in ribosome biogenesis and contains an acidic N terminus that binds nuclear localization sequences. In previous work we showed that human nucleolin associates with the N-terminal region of human topoisomerase I (Top1). We have now mapped the topoisomerase I interaction domain of nucleolin to the N-terminal 225 amino acids. We also show that the Saccharomyces cerevisiae nucleolin ortholog, Nsr1p, physically interacts with yeast topoisomerase I, yTop1p. Studies of isogenic NSR1(+) and Deltansr1 strains indicate that NSR1 is important in determining the cellular localization of yTop1p. Moreover, deletion of NSR1 reduces sensitivity to camptothecin, an antineoplastic topoisomerase I inhibitor. By contrast, Deltansr1 cells are hypersensitive to the topoisomerase II-targeting drug amsacrine. These findings indicate that nucleolin/Nsr1 is involved in the cellular localization of Top1 and that this localization may be important in determining sensitivity to drugs that target topoisomerases.     

Preliminary Evaluation of a Bunyavirus Vector for Cancer Immunotherapy

Replicon particles of Rift Valley fever virus, referred to as nonspreading Rift Valley fever virus (NSR), are intrinsically safe and highly immunogenic. Here, we demonstrate that NSR-infected human dendritic cells can activate CD8⁺ T cells in vitro and that prophylactic and therapeutic vaccinations of mice with NSR encoding a tumor-associated CD8 peptide can control the outgrowth of lymphoma cells in vivo. These results suggest that the NSR system holds promise for cancer immunotherapy.     

Localization of insulin-like immunoreactivity in the synganglion of nymphal and adult Dermacentor variabilis (Acari: Ixodidae)

Immunocytochemical staining based on a peroxidase-antiperoxidase method showed neurosecretory cells (NSC) reactive to bovine insulin in five of 18 paraldehyde fuchsin-positive neurosecretory regions (NSR) in the synganglion of unfed adult Dermacentor variabilis. This is the first report of a neuropeptide in an ixodid tick. The insulin-specific immunoreactive cells included the posterior medial group of the protocerebral center, posterior group of dorsal opisthosomal center, anterior lateral group of the dorso-lateral cheliceral center, dorsal group of the frontal stomodeal center, and anterior group of the ventral palpal center. After feeding and mating, females no longer had immunoreactive cells in three of five NSR found in virgin, unfed females. However, two cells of the posterior group in dorsal opisthosomal center and anterior lateral group of the dorso-lateral cheliceral center remained immunoreactive throughout feeding. Fed, mated males continued to display immunoreactive cells in four of five NSR found in the virgin, unfed males. All developmental stages of nymphs examined had insulin-specitic immunoreactive cells in two of the five NSR found in unfed adults, including two positively stained cells of the posterior group in dorsal opisthosomal center and anterior group of ventral palpal neurosecretory center.     

Insights into the metabolism of elemental sulfur by the hyperthermophilic archaeon Pyrococcus furiosus: characterization of a coenzyme A- dependent NAD(P)H sulfur oxidoreductase

The hyperthermophilic archaeon Pyrococcus furiosus uses carbohydrates as a carbon source and produces acetate, CO2, and H2 as end products. When S(0) is added to a growing culture, within 10 min the rate of H2 production rapidly decreases and H(2)S is detected. After 1 hour cells contain high NADPH- and coenzyme A-dependent S(0) reduction activity (0.7 units/mg, 85 degrees C) located in the cytoplasm. The enzyme responsible for this activity was purified to electrophoretic homogeneity (specific activity, 100 units/mg) and is termed NAD(P)H elemental sulfur oxidoreductase (NSR). NSR is a homodimeric flavoprotein (M(r), 100,000) and is encoded by PF1186. This designation was previously assigned to the gene encoding an enzyme that reduces coenzyme A disulfide, which is a side reaction of NSR. Whole-genome DNA microarray and quantitative PCR analyses showed that the expression of NSR is up-regulated up to sevenfold within 10 min of S(0) addition. This primary response to S(0) also involves the up-regulation (>16-fold) of a 13-gene cluster encoding a membrane-bound oxidoreductase (MBX). The cluster encoding MBX is proposed to replace the homologous 14-gene cluster that encodes the ferredoxin-oxidizing, H2-evolving membrane-bound hydrogenase (MBH), which is down-regulated >12-fold within 10 min of S(0) addition. Although an activity for MBX could not be demonstrated, it is proposed to conserve energy by oxidizing ferredoxin and reducing NADP, which is used by NSR to reduce S(0). A secondary response to S(0) is observed 30 min after S(0) addition and includes the up-regulation of genes encoding proteins involved in amino acid biosynthesis and iron metabolism, as well as two so-called sulfur-induced proteins termed SipA and SipB. This novel S(0)-reducing system involving NSR and MBX has been found so far only in the heterotrophic Thermococcales and is in contrast to the cytochrome- and quinone-based S(0)-reducing system in autotrophic archaea and bacteria.     

China's U-Shaped Line in the South China Sea Revisited

Despite its existence on the Chinese maps for more than six decades, the U-shaped line, as a traditional maritime boundary line of China in the South China Sea, has never received a wide recognition in the world community, much less by the other claimant states in the South China Sea. The U-shaped line is a legal conundrum not only for China but also for the world community, particularly after the map with the U-shaped line, together with China's Notes Verbale with respect to the claims to the outer continental shelves made by Malaysia and Vietnam, were submitted to the UN Commission on the Limits of Continental Shelf in May 2009. This article discusses China's recent practice relating to the U-shaped line as well as the external factors that affect the validity of the line and tries to unravel the legal puzzle posed by the line.     

Protection of the antarctic ecosystem: A study in international environmental law

Abstract

The Antarctic region constitutes a fragile eco‐system closely related to the unique features of the physical environment of that continent. The Antarctic Ocean is central to the region's living systems, with krill as the ecological basis of life in the ocean and on land. So far, man's impact upon the Antarctic environment has been negligible, but there is concern that overharvesting of krill and possible marine pollution resulting from any future offshore oil exploration may undermine the krill basis of the Antarctic ecosystem. The legal status of Antarctica is largely determined by the 1959 Antarctic Treaty, and especially by the inner circle of the currently fourteen “consultative”; status signatories. These states have given high priority to ecological considerations by enacting a series of environmental and conservationist regulations, as well as two conventions, one protecting the Antarctic seals and the other marine living resources in general. Environmental regulations will form an important part of the legal regime for the exploration and exploitation of the Antarctic mineral resources, primarily oil and gas. The Antarctic Treaty framework does not legally bind nonsignatory states, but under general international law all states are bound to refrain from inflicting damage upon the planet's environment. Also, some marine pollution conventions apply to the Antarctic waters, and the 1982 U.N. Convention on the Law of the Sea comprehensively covers the protection and preservation of the marine environment of all oceans and seas. The preservation of the Antarctic environment will remain a high priority irrespective of what legal regime will govern Antarctica after 1991 when the Antarctic Treaty may, and most probably will, be subject to review.