Proteasome inhibitors increase missense mutated dysferlin in patients with muscular dystrophy

No treatment is available for patients affected by the recessively inherited, progressive muscular dystrophies caused by a deficiency in the muscle membrane repair protein dysferlin. A marked reduction in dysferlin in patients harboring missense mutations in at least one of the two pathogenic DYSF alleles encoding dysferlin implies that dysferlin is degraded by the cell's quality control machinery. In vitro evidence suggests that missense mutated dysferlin might be functional if salvaged from degradation by the proteasome. We treated three patients with muscular dystrophy due to a homozygous Arg555Trp mutation in dysferlin with the proteasome inhibitor bortezomib and monitored dysferlin expression in monocytes and in skeletal muscle by repeated percutaneous muscle biopsy. Expression of missense mutated dysferlin in the skeletal muscle and monocytes of the three patients increased markedly, and dysferlin was correctly localized to the sarcolemma of muscle fibers on histological sections. Salvaged missense mutated dysferlin was functional in a membrane resealing assay in patient-derived muscle cells treated with three different proteasome inhibitors. We conclude that interference with the proteasomal system increases expression of missense mutated dysferlin, suggesting that this therapeutic strategy may benefit patients with dysferlinopathies and possibly other genetic diseases.

Heavy metal and petroleum product concentrations in waters at the Obskaya 1 drill site

During recent years, the basins of the Kara Sea (Kamennomysskaya, Obskaya, and Chugor'yakhinskaya structures) in the Russian Federation have been considered as promising regions for oil and gas exploration and, simultaneously, as possible paths of relatively cheap pipeline and tanker transportation of hydrocarbons projected for recovery. On the other hand, exploration operations, recovery, and transportation of gas pose a considerable risk of accidents and environmental pollution, which causes a justified concern about the future state of the ecological system of the Gulf of Ob and the adjoining parts of the Kara Sea. Therefore, regular combined environmental investigations (monitoring) are the most important factor for estimating the current state and forecasting the dynamics of the development of estuary systems. The program of investigations (schedule, station network, and measured parameters) is standardized in accordance with the international practice of such work and accounts for the experience of monitoring studies of Russian and foreign researchers. Two measurement sessions were performed during ecological investigations in the region of exploration drilling: at the beginning at final stage of drilling operations and borehole testing; in addition, natural parameters were determined in various parts of the Ob estuary before the beginning of investigations. Hydrophysical and hydrochemical characteristics of the water medium were determined and bottom sediments and water were analyzed for various pollutants (petroleum products, heavy metals, and radionuclides). The forms of heavy-metal occurrence in river and sea waters were determined by the method of continuous multistep filtration, which is based on water component fractionation on membrane filters of various pore sizes. These investigations revealed environmental pollution by chemical substances during the initial stage of drilling operations, when remains of fuels, oils, and solutions could be spilled, and part of the chemical pollutants could enter the environment. Owing to horizontal and vertical turbulent diffusion, wave mixing, and the effect of the general direction of currents in the Ob estuary from south to north, areas are formed with elevated concentrations of the analyzed elements and compounds. However, the concentration levels of chemical pollutants are practically no higher than the maximum admissible concentrations, and their substantial dissipation to the average regional background contents can be expected in the near future. Our investigations allowed us to determine in detail the parameters of anthropogenic pollution in the regions affected by hydrocarbon exploration drilling in the Obskii and Kamennomysskii prospects in the Gulf of Ob and estimate their influence on the ecological state of the basin of the Ob River and the Kara Sea on the whole.

Benthic foraminifera and stable isotope composition in Paleocene-Eocene sediments

In the late Paleocene to early Eocene, deep sea benthic foraminifera suffered their only global extinction of the last 75 million years and diversity decreased worldwide by 30-50% in a few thousand years. At Maud Rise (Weddell Sea, Antarctica; Sites 689 and 690, palaeodepths 1100 m and 1900 m) and Walvis Ridge (Southeastern Atlantic, Sites 525 and 527, palaeodepths 1600 m and 3400 m) post-extinction faunas were low-diversity and high-dominance, but the dominant species differed by geographical location. At Maud Rise, post-extinction faunas were dominated by small, biserial and triserial species, while the large, thick-walled, long-lived deep sea species Nuttallides truempyi was absent. At Walvis Ridge, by contrast, they were dominated by long-lived species such as N. truempyi, with common to abundant small abyssaminid species. The faunal dominance patterns at the two locations thus suggest different post-extinction seafloor environments: increased flux of organic matter and possibly decreased oxygen levels at Maud Rise, decreased flux at Walvis Ridge. The species-richness remained very low for about 50 000 years, then gradually increased. The extinction was synchronous with a large, negative, short-term excursion of carbon and oxygen isotopes in planktonic and benthic foraminifera and bulk carbonate. The isotope excursions reached peak negative values in a few thousand years and values returned to pre-excursion levels in about 50 000 years. The carbon isotope excursion was about -2 per mil for benthic foraminifera at Walvis Ridge and Maud Rise, and about -4 per mil for planktonic foraminifera at Maud Rise. At the latter sites vertical gradients thus decreased, possibly at least partially as a result of upwelling. The oxygen isotope excursion was about -1.5 per mil for benthic foraminifera at Walvis Ridge and Maud Rise, -1 per mil for planktonic foraminifera at Maud Rise. The rapid oxygen isotope excursion at a time when polar ice-sheets were absent or insignificant can be explained by an increase in temperature by 4-6°C of high latitude surface waters and deep waters world wide. The deep ocean temperature increase could have been caused by warming of surface waters at high latitudes and continued formation of the deep waters at these locations, or by a switch from dominant formation of deep waters at high latitudes to formation at lower latitudes. Benthic foraminiferal post-extinction biogeographical patterns favour the latter explanation. The short-term carbon isotope excursion occurred in deep and surface waters, and in soil concretions and mammal teeth in the continental record. It is associated with increased CaC03-dissolution over a wide depth range in the oceans, suggesting that a rapid transfer of isotopically light carbon from lithosphere or biosphere into the ocean-atmosphere system may have been involved. The rapidity of the initiation of the excursion (a few thousand years) and its short duration (50 000 years) suggest that such a transfer was probably not caused by changes in the ratio of organic carbon to carbonate deposition or erosion. Transfer of carbon from the terrestrial biosphere was probably not the cause, because it would require a much larger biosphere destruction than at the end of the Cretaceous, in conflict with the fossil record. It is difficult to explain the large shift by rapid emission into the atmosphere of volcanogenic CO2, although huge subaerial plateau basalt eruptions occurred at the time in the northern Atlantic. Probably a complex combination of processes and feedback was involved, including volcanogenic emission of CO2, changing circulation patterns, changing productivity in the oceans and possibly on land, and changes in the relative size of the oceanic and atmospheric carbon reservoirs.

Industrialization in Indonesia since the 1970s

Introduction: During the period from the latter half of the 1980s until just before the Asian currency crisis in 1997, Indonesia’s economic development had drawn expectations and attention from various quarters, along with Malaysia and Thailand within the same Association of Southeast Asian Nations (ASEAN). In fact, the 1993 report by the World Bank, entitled “East Asian Miracle: Economic Growth and Public Policy,” recognized Indonesia as one of the East Asian economies with the strong economic performance, i.e. sustained economic growth (World Bank [1993]). And it was the manufacturing industry that had been the driving force behind Indonesia’s economic growth during that period. Since the 1997 outbreak of the Asian currency crisis, however, the manufacturing sector in Indonesia has been mired in a situation that rules out the kind of bright prospects it had emanated previously. The Indonesian economy is still in the developing stage, and in accordance with the history of industrial structural changes in other countries, Indonesia’s manufacturing industry can still be expected to serve as the engine of the country’s economic development. But is it really possible in an environment where economic liberalization and globalization are forging ahead? And, what sort of problems have to be dealt with to make it possible? To answer these questions, it is necessary to know the current conditions of Indonesia’s manufacturing sector, and to do that, it becomes important to think back on the history of the country’s industrialization. Thus, this paper is intended to retrace and unlock the track of Indonesia’s industrialization up until the establishment of the manufacturing sector in its present form, with the ultimate goal being to give answers to the above-mentioned questions. Subject to an analysis in this paper is the period from the installment of President Soeharto’s administration onward when industrialization of the modern industrial sector2 moved into high gear. 　　　The composition of this paper is outlined below. Section 1 first shows why it is important to examine import substitution and export orientation, both of which are used as the measures of the analysis in this paper, in tracking the history of the industrialization, and then discuss indicators of import substitution and export orientation as well as statistical data and resources needed to develop those indicators. Section 2 clarifies the status of the manufacturing industry among all industries by looking at the composition ratio of the manufacturing industry in terms of value added, imports and exports. Section 3 to 5 cover three periods between 1971 and 1995 and make an analysis of import substitution, export orientation and changes in the industrial structure for each period. Section 3 analyzes the period from 1971 through 1985, when Indonesia pursued the import substitution policy amid the oil boom. Section 4 covers the period from 1985 through 1990, when the packages of deregulatory measures were announced successively under structural adjustment policies made necessary by the fall in oil prices. Section 5 examines the period from 1990 through 1995, which saw the alternate shifts between the overheating of the economy by sharply rising investment by both domestic and foreign investors in the wake of the liberalization of investment, trade and financial services, and polices to cool down the economy. Section 6, which covers the 1995-1999 period straddling the economic crisis, is designed for an analysis of the changes in production trends before and after the economic crisis as well as the changes in the industrial structure. Section 7, after summing up the history of Indonesia’s industrialization examined in the previous sections, discusses problems found in respective sectors and attempts to present future prospects for the country’s manufacturing industry.

Research lines in Hyperthermia at the Bioinstrumentation Laboratory of the Centre for Biomedical Technology

The Bioinstrumentation Laboratory belongs to the Centre for Biomedical Technology (CTB) of the Technical University of Madrid and its main objective is to provide the scientific community with devices and techniques for the characterization of micro and nanostructures and consequently finding their best biomedical applications. Hyperthermia (greek word for “overheating”) is defined as the phenomenon that occurs when a body is exposed to an energy generating source that can produce a rise in temperature (42-45ºC) for a given time [1]. Specifically, the aim of the hyperthermia methods used in The Bioinstrumentation Laboratory is the development of thermal therapies, some of these using different kinds of nanoparticles, to kill cancer cells and reduce the damage on healthy tissues. The optical hyperthermia is based on noble metal nanoparticles and laser irradiation. This kind of nanoparticles has an immense potential associated to the development of therapies for cancer on account of their Surface Plasmon Resonance (SPR) enhanced light scattering and absorption. In a short period of time, the absorbed light is converted into localized heat, so we can take advantage of these characteristics to heat up tumor cells in order to obtain the cellular death [2]. In this case, the laboratory has an optical hyperthermia device based on a continuous wave laser used to kill glioblastoma cell lines (1321N1) in the presence of gold nanorods (Figure 1a). The wavelength of the laser light is 808 nm because the penetration of the light in the tissue is deeper in the Near Infrared Region. The first optical hyperthermia results show that the laser irradiation produces cellular death in the experimental samples of glioblastoma cell lines using gold nanorods but is not able to decrease the cellular viability of cancer cells in samples without the suitable nanorods (Figure 1b) [3]. The generation of magnetic hyperthermia is performed through changes of the magnetic induction in magnetic nanoparticles (MNPs) that are embedded in viscous medium. The Figure 2 shows a schematic design of the AC induction hyperthermia device in magnetic fluids. The equipment has been manufactured at The Bioinstrumentation Laboratory. The first block implies two steps: the signal selection with frequency manipulation option from 9 KHz to 2MHz, and a linear output up to 1500W. The second block is where magnetic field is generated ( 5mm, 10 turns). Finally, the third block is a software control where the user can establish initial parameters, and also shows the temperature response of MNPs due to the magnetic field applied [4-8]. The Bioinstrumentation Laboratory in collaboration with the Mexican company MRI-DT have recently implemented a new research line on Nuclear Magnetic Resonance Hyperthermia, which is sustained on the patent US 7,423,429B2 owned by this company. This investigation is based on the use of clinical MRI equipment not only for diagnosis but for therapy [9]. This idea consists of two main facts: Magnetic Resonance Imaging can cause focal heating [10], and the differentiation in resonant frequency between healthy and cancer cells [11]. To produce only heating in cancer cells when the whole body is irradiated, it is necessary to determine the specific resonant frequency of the target, using the information contained in the spectra of the area of interest. Then, special RF pulse sequence is applied to produce fast excitation and relaxation mechanism that generates temperature increase of the tumor, causing cellular death or metabolism malfunction that stops cellular division

GIPC, a PDZ domain containing protein, interacts specifically with the C terminus of RGS-GAIP

We have identified a mammalian protein called GIPC (for GAIP interacting protein, C terminus), which has a central PDZ domain and a C-terminal acyl carrier protein (ACP) domain. The PDZ domain of GIPC specifically interacts with RGS-GAIP, a GTPase-activating protein (GAP) for Gαi subunits recently localized on clathrin-coated vesicles. Analysis of deletion mutants indicated that the PDZ domain of GIPC specifically interacts with the C terminus of GAIP (11 amino acids) in the yeast two-hybrid system and glutathione S-transferase (GST)-GIPC pull-down assays, but GIPC does not interact with other members of the RGS (regulators of G protein signaling) family tested. This finding is in keeping with the fact that the C terminus of GAIP is unique and possesses a modified C-terminal PDZ-binding motif (SEA). By immunoblotting of membrane fractions prepared from HeLa cells, we found that there are two pools of GIPC–a soluble or cytosolic pool (70%) and a membrane-associated pool (30%). By immunofluorescence, endogenous and GFP-tagged GIPC show both a diffuse and punctate cytoplasmic distribution in HeLa cells reflecting, respectively, the existence of soluble and membrane-associated pools. By immunoelectron microscopy the membrane pool of GIPC is associated with clusters of vesicles located near the plasma membrane. These data provide direct evidence that the C terminus of a RGS protein is involved in interactions specific for a given RGS protein and implicates GAIP in regulation of additional functions besides its GAP activity. The location of GIPC together with its binding to GAIP suggest that GAIP and GIPC may be components of a G protein-coupled signaling complex involved in the regulation of vesicular trafficking. The presence of an ACP domain suggests a putative function for GIPC in the acylation of vesicle-bound proteins.

Yeast Dam1p Is Required to Maintain Spindle Integrity during Mitosis and Interacts with the Mps1p Kinase

We have identified a mutant allele of the DAM1 gene in a screen for mutations that are lethal in combination with the mps1-1 mutation. MPS1 encodes an essential protein kinase that is required for duplication of the spindle pole body and for the spindle assembly checkpoint. Mutations in six different genes were found to be lethal in combination with mps1-1, of which only DAM1 was novel. The remaining genes encode a checkpoint protein, Bub1p, and four chaperone proteins, Sti1p, Hsc82p, Cdc37p, and Ydj1p. DAM1 is an essential gene that encodes a protein recently described as a member of a microtubule binding complex. We report here that cells harboring the dam1-1 mutation fail to maintain spindle integrity during anaphase at the restrictive temperature. Consistent with this phenotype, DAM1 displays genetic interactions with STU1, CIN8, and KAR3, genes encoding proteins involved in spindle function. We have observed that a Dam1p-Myc fusion protein expressed at endogenous levels and localized by immunofluorescence microscopy, appears to be evenly distributed along short mitotic spindles but is found at the spindle poles at later times in mitosis.

Class I release factors in ciliates with variant genetic codes

In eukaryotes with the universal genetic code a single class I release factor (eRF1) most probably recognizes all stop codons (UAA, UAG and UGA) and is essential for termination of nascent peptide synthesis. It is well established that stop codons have been reassigned to amino acid codons at least three times among ciliates. The codon specificities of ciliate eRF1s must have been modified to accommodate the variant codes. In this study we have amplified, cloned and sequenced eRF1 genes of two hypotrichous ciliates, Oxytricha trifallax (UAA and UAG for Gln) and Euplotes aediculatus (UGA for Cys). We also sequenced/identified three protist and two archaeal class I RF genes to enlarge the database of eRF1/aRF1s with the universal code. Extensive comparisons between universal code eRF1s and those of Oxytricha, Euplotes and Tetrahymena, which represent three lineages that acquired variant codes independently, provide important clues to identify stop codon-binding regions in eRF1. Domain 1 in the five ciliate eRF1s, particulary the TASNIKS heptapeptide and its adjacent region, differs significantly from domain 1 in universal code eRF1s. This observation suggests that domain 1 contains the codon recognition site, but that the mechanism of eRF1 codon recognition may be more complex than proposed by Nakamura et al. or Knight and Landweber.

ADP-Dependent Phosphorylation Regulates Association of a DNA-Binding Complex with the Barley Chloroplast psbD Blue-Light-Responsive Promoter1

The chloroplast gene psbD encodes D2, a chlorophyll-binding protein located in the photosystem II reaction center. Transcription of psbD in higher plants involves at least three promoters, one of which is regulated by blue light. The psbD blue-light-regulated promoter (BLRP) consists of a −10 promoter element and an activating complex, AGF, that binds immediately upstream of −35. A second sequence-specific DNA-binding complex, PGTF, binds upstream of AGF between −71 and −100 in the barley (Hordeum vulgare) psbD BLRP. In this study we report that ADP-dependent phosphorylation selectively inhibits the binding of PGTF to the barley psbD BLRP. ATP at high concentrations (1–5 mm) inhibits PGTF binding, but in the presence of phosphocreatine and phosphocreatine kinase, this capacity is lost, presumably due to scavenging of ADP. ADP inhibits PGTF binding at relatively low concentrations (0.1 mm), whereas other nucleotides are unable to mediate this response. ADP-mediated inhibition of PGTF binding is reduced in the presence of the protein kinase inhibitor K252a. This and other results suggest that ADP-dependent phosphorylation of PGTF (or some associated protein) inhibits binding of PGTF to the psbD BLRP and reduces transcription. ADP-dependent phosphorylation is expected to increase in darkness in parallel with the rise in ADP levels in chloroplasts. ADP-dependent phosphorylation in chloroplasts may, therefore, in coordination, inactivate enzymes involved in carbon assimilation, protein synthesis, and transcription during diurnal light/dark cycles.

The replication initiator protein pi of the plasmid R6K specifically interacts with the host-encoded helicase DnaB.

The replication initiator protein pi of plasmid R6K is known to interact with the seven iterons of the gamma origin/enhancer and activate distant replication origins alpha and beta (ori alpha and ori beta) by pi-mediated DNA looping. Here we show that pi protein specifically interacts in vitro with the host-encoded helicase DnaB. The site of interaction of pi on DnaB has been localized to a 37-aa-long region located between amino acids 151 and 189 of DnaB. The surface of pi that interacts with DnaB has been mapped to the N-terminal region of the initiator protein between residues 1 and 116. The results suggest that during initiation of replication, the replicative helicase DnaB is first recruited to the gamma enhancer by the pi protein. In a subsequent step, the helicase probably gets delivered from ori gamma to ori alpha and ori beta by pi-mediated DNA looping.

Injury incidence in judokas at the Spanish National University Championship

Background and Study Aim: Understanding injury incidence rates will be a great help with regards to preventing potential future damages. It is for this reason that this study suggests studying a large number of variables. The purpose of research is the relationship of events (empirical variables) that are usually taken into account in developing injury prevention programs during the battles and training in judo tournament. Material and methods: In this research project, 57 male judokas taking part in the Spanish National University Championship in 2009 were asked to complete a retrospective questionnaire. We analysed the following events: the most commonly injured body regions, the medical diagnosis, how and when the injury happened, the type of injury, the side of the body and the type of medical attention received. For the statistical analysis, we used the SPSS statistics programme to apply the Chi-square test in order to determine the significance levels for non-parametric tests from p<.05. Results: Significant differences were found in the most commonly injured body region, the shoulder/clavicle (p<.05), and in the most common diagnosis, the sprain (p<.05). Impact injuries (p<.05) are the most common and training (p<.05) is the most dangerous time. About the type of injury, 78.38% are new injuries (p<.05) and 69.05% affect the right hand side of the body (p<.05). Doctors are the most consulted specialists, but the physiotherapists obtained the best marks. Have been out due to injury for over 21 days 36.36% of the participants, but not for the entire season. Conclusions: The most common diagnosis in university student judokas coincides with those of elite judokas, with the sprain being the most common. University student judokas have a higher rate of shoulder/clavicle injuries, while professional judokas are prone to a higher rate of knee injuries. Training is the most common moment in which injuries occur, both in university student judokas and professional judokas. New injuries are the most common types of injuries in university student judokas and, while doctors are the most consulted specialists, the physiotherapists obtained the best marks.

Satisfaction of clinical nurses with the formation of a university tutorial program

Degree in nursing from the Universitat Jaume I (UJI) maintains the continuity of learning with an integrated learning methodology (theory, simulated practice and clinical practice). The objective of this methodology is to achieve consistency between the knowledge, abilities and skills acquired in the classroom, laboratory and clinic to ensure skills related. Reference Nurse is a key figure in this process, you receive accredited training on Educational Methods, assessment of competence, and Evidence-Based Practice that plays the role of evaluating in conjunction with the subjects. It does not perceive economic remuneration. The main objective of this study is to determine the level of satisfaction of clinical nurses on the Nurses Training Program Reference in UJI (Castellon- Spain). A cross sectional study was performed and conducted on 150 nurses. 112 questionnaires were completed, collected and analysed at the end of training. The survey consists of 12 items measured with the Likert scale with 5 levels of response and two open questions regarding the positive and negative aspects of the course and to add in this formation. The training is always performed by the same faculty and it's used four sessions of 2012. We perform a quantitative analysis of the variables under study using measures of central tendency. The completion rate of the survey is 95.53% (n=107). Anonymity rate of 54,14% The overall satisfaction level of training was 3.65 (SD = 0.89) on 5 points. 54.2% (n = 58) of the reference nurses made a contribution in the open questions described in the overall results. The overall satisfaction level can be considered acceptable. It is considered necessary to elaborate a specific survey to detect areas of improvement of nurse training program reference and future recruitment strategies. The main objective of the present work is the selection and integration of different methodologies among those applicable within the framework of the European Higher Education Area to combine teaching methods with high implication from both lecturers and students.

Abkhazia, South Ossetia, Nagorno-Karabakh: unfrozen conflicts between Russia and the West. OSW Special Report, 9 July 2008

The Southern Caucasus is the site of three armed conflicts with separatist backgrounds, which have remained unsolved for years: the conflicts in Georgia's Abkhazia and South Ossetia, and Azerbaijan's conflict over Nagorno-Karabakh (including the areas around Nagorno-Karabakh which were seized by Armenian separatists in the course of the war). Neither Georgia nor Azerbaijan have had any control over the disputed areas since the early 1990s. Both states are simultaneously in conflict with the separatists' informal patrons, respectively Russia and Armenia. After over a decade of relative peace during which the conflicts remained frozen, tension has recently risen considerably: in the case of Abkhazia and South Ossetia, large-scale fighting may break out in the coming months, whereas in the case of Nagorno-Karabakh and the Azeri-Armenian conflict, such a threat may materialise within the next five years. The current formula for politically resolving the conflicts is ineffective and close to exhaustion, and the prospect of any alternative peace plans being developed is rather distant. The conflicts in the Southern Caucasus are of increasing concern to the West, mainly because of the Western actors' constantly growing political and economic involvement in Georgia and Azerbaijan (including support for reforms and development of the gas and oil transmission infrastructures), as well as its less intensive commitments in Armenia. An outbreak of open fighting over the separatist regions would destabilise the Southern Caucasus, largely undoing the results of the actions which the EU, NATO and the USA have taken in the region in recent years. Moreover, the situation in the Southern Caucasus, especially the separatisms themselves, have in fact become an element in the wider geopolitical game between the West and Russia. For Russia, the stakes are maintaining its influence in the region, and for the West, demonstrating its ability to effectively promote democracy and economic modernisation in the countries bordering it.

Economic relations with the U.S.S.R.,

At head of title: The Committee on International Economic Policy in cooperation with the Carnegie Endowment for International Peace.

Message from the President of the United States transmitting a report of the Secretary of agriculture in relation to the forests, rivers, and mountains of the Southern Appalachian region. December 19, 1901. -- Read, referred to the Committee on forest reservations and the protection of game and ordered to be printed.

Prepared in cooperation with the Department of the Interior. Appendix A is submitted by the Bureau of forestry, B-C by the Geological survey and D by the Weather bureau.