Excitation–contraction coupling is unaffected by drastic alteration of the sequence surrounding residues L720–L764 of the α1S II-III loop

The II-III loop of the skeletal muscle dihydropyridine receptor (DHPR) α1S subunit is responsible for bidirectional-signaling interactions with the ryanodine receptor (RyR1): transmitting an orthograde, excitation–contraction (EC) coupling signal to RyR1 and receiving a retrograde, current-enhancing signal from RyR1. Previously, several reports argued for the importance of two distinct regions of the skeletal II-III loop (residues R681–L690 and residues L720–Q765, respectively), claiming for each a key function in DHPR–RyR1 communication. To address whether residues 720–765 of the II-III loop are sufficient to enable skeletal-type (Ca2+ entry-independent) EC coupling and retrograde interaction with RyR1, we constructed a green fluorescent protein (GFP)-tagged chimera (GFP-SkLM) having rabbit skeletal (Sk) DHPR sequence except for a II-III loop (L) from the DHPR of the house fly, Musca domestica (M). The Musca II-III loop (75% dissimilarity to α1S) has no similarity to α1S in the regions R681–L690 and L720–Q765. GFP-SkLM expressed in dysgenic myotubes (which lack endogenous α1S subunits) was unable to restore EC coupling and displayed strongly reduced Ca2+ current densities despite normal surface expression levels and correct triad targeting (colocalization with RyR1). Introducing rabbit α1S residues L720–L764 into the Musca II-III loop of GFP-SkLM (substitution for Musca DHPR residues E724–T755) completely restored bidirectional coupling, indicating its dependence on α1S loop residues 720–764 but its independence from other regions of the loop. Thus, 45 α1S-residues embedded in a very dissimilar background are sufficient to restore bidirectional coupling, indicating that these residues may be a site of a protein–protein interaction required for bidirectional coupling.

Ethidium-dependent uncoupling of substrate binding and cleavage by Escherichia coli ribonuclease III

Ethidium bromide (EB) is known to inhibit cleavage of bacterial rRNA precursors by Escherichia coli ribonuclease III, a dsRNA-specific nuclease. The mechanism of EB inhibition of RNase III is not known nor is there information on EB-binding sites in RNase III substrates. We show here that EB is a reversible, apparently competitive inhibitor of RNase III cleavage of small model substrates in vitro. Inhibition is due to intercalation, since (i) the inhibitory concentrations of EB are similar to measured EB intercalation affinities; (ii) substrate cleavage is not affected by actinomycin D, an intercalating agent that does not bind dsRNA; (iii) the EB concentration dependence of inhibition is a function of substrate structure. In contrast, EB does not strongly inhibit the ability of RNase III to bind substrate. EB also does not block substrate binding by the C-terminal dsRNA-binding domain (dsRBD) of RNase III, indicating that EB perturbs substrate recognition by the N-terminal catalytic domain. Laser photocleavage experiments revealed two ethidium-binding sites in the substrate R1.1 RNA. One site is in the internal loop, adjacent to the scissile bond, while the second site is in the lower stem. Both sites consist of an A-A pair stacked on a CG pair, a motif which apparently provides a particularly favorable environment for intercalation. These results indicate an inhibitory mechanism in which EB site-specifically binds substrate, creating a cleavage-resistant complex that can compete with free substrate for RNase III. This study also shows that RNase III recognition and cleavage of substrate can be uncoupled and supports an enzymatic mechanism of dsRNA cleavage involving cooperative but not obligatorily linked actions of the dsRBD and the catalytic domain.

Rapid refolding of a proline-rich all-beta-sheet fibronectin type III module.

Fibronectin type III modules contain approximately 90 residues and are an extremely common building block of animal proteins. Despite containing a complex all-beta-sheet topology and eight prolines, the refolding of the 10th type III module of human fibronectin has been found to be very rapid, with native core packing, amide hydrogen bonding, and backbone conformation all recovered within 1 s at 5 degrees C. These observations indicate that this domain can overcome many structural characteristics often thought to slow the folding process.

The SNAP45 subunit of the small nuclear RNA (snRNA) activating protein complex is required for RNA polymerase II and III snRNA gene transcription and interacts with the TATA box binding protein.

The RNA polymerase II and III small nuclear RNA (snRNA) promoters contain a common basal promoter element, the proximal sequence element (PSE). The PSE binds a multisubunit complex we refer to as the snRNA activating protein complex (SNAPc). At least four polypeptides are visible in purified SNAPc preparations, which migrate with apparent molecular masses of 43, 45, 50, and 190 kDa on SDS/polyacrylamide gels. In addition, purified preparations of SNAPc contain variable amounts of TATA box binding protein (TBP). An important question is whether the PSEs of RNA polymerase II and III snRNA promoters recruit the exact same SNAP complex or slightly different versions of SNAPc, differing, for example, by the presence or absence of a subunit. To address this question, we are isolating cDNAs encoding different subunits of SNAPc. We have previously isolated the cDNA encoding the 43-kDa subunit SNAP43. We now report the isolation of the cDNA that encodes the p45 polypeptide. Antibodies directed against p45 retard the mobility of the SNAPc-PSE complex in an electrophoretic mobility shift assay, indicating that p45 is indeed part of SNAPc. We therefore refer to this protein as SNAP45. SNAP45 is exceptionally proline-rich, interacts strongly with TBP, and, like SNAP43, is required for both RNA polymerase II and III transcription of snRNA genes.

Inequality in analgesic prescription in Spain: a gender development issue

Objectives: It is well known that sex differences in analgesic prescription are not merely the logical result of greater prevalence of pain in women, since this therapeutic variability is related to factors such as educational level or social class. This study aims to analyse the relationship between analgesic prescription and gender development in different regions of Spain. Methods: Cross-sectional study of sex-differences in analgesic prescription according to the gender development of the regions studied. Analgesic prescription, pain and demographic variables were obtained from the Spanish Health Interview Survey in 2006. Gender development was measured with the Gender Development Index (GDI). A logistic regression analysis was conducted to compare analgesic prescription by sex in regions with a GDI above or below the Spanish average. Results: Once adjusted by pain, age and social class, women were more likely to be prescribed analgesics than men, odds ratio (OR) = 1.74 (1.59-1.91), as residents in regions with a lower GDI compared with those in region with a higher GDI: ORWomen = 1.26 (1.12-1.42), ORMen = 1.30 (1.13-1.50). Women experiencing pain in regions with a lower GDI were more likely than men to be treated by a general practitioner rather than by a specialist, OR = 1.32 (1.04-1.67), irrespective of age and social class. Conclusions: Gender bias may be one of the pathways by which inequalities in analgesic treatment adversely affect women's health. Moreover, research into the adequacy of analgesic treatment and the possible medicalisation of women should consider contextual factors, such as gender development.

Biodiversity of Archaea and floral of two inland saltern ecosystems in the Alto Vinalopó Valley, Spain

Background. The extraction of salt from seawater by means of coastal solar salterns is a very well-described process. Moreover, the characterization of these environments from ecological, biochemical and microbiological perspectives has become a key focus for many research groups all over the world over the last 20 years. In countries such as Spain, there are several examples of coastal solar salterns (mainly on the Mediterranean coast) and inland solar salterns, from which sodium chloride is obtained for human consumption. However, studies focused on the characterization of inland solar salterns are scarce and both the archaeal diversity and the plant communities inhabiting these environments remain poorly described. Results. Two of the inland solar salterns (termed Redonda and Penalva), located in the Alto Vinalopó Valley (Alicante, Spain), were characterized regarding their geological and physico-chemical characteristics and their archaeal and botanical biodiversity. A preliminary eukaryotic diversity survey was also performed using saline water. The chemical characterization of the brine has revealed that the salted groundwater extracted to fill these inland solar salterns is thalassohaline. The plant communities living in this environment are dominated by Sarcocornia fruticosa (L.) A.J. Scott, Arthrocnemum macrostachyum (Moris) K. Koch, Suaeda vera Forsk. ex Gmelin (Amaranthaceae) and several species of Limonium (Mill) and Tamarix (L). Archaeal diversity was analyzed and compared by polymerase chain reaction (PCR)-based molecular phylogenetic techniques. Most of the sequences recovered from environmental DNA samples are affiliated with haloarchaeal genera such as Haloarcula, Halorubrum, Haloquadratum and Halobacterium, and with an unclassified member of the Halobacteriaceae. The eukaryote Dunaliella was also present in the samples. Conclusions. To our knowledge, this study constitutes the first analysis centered on inland solar salterns located in the southeastern region of Spain. The results obtained revealed that the salt deposits of this region have marine origins. Plant communities typical of salt marshes are present in this ecosystem and members of the Halobacteriaceae family can be easily detected in the microbial populations of these habitats. Possible origins of the haloarchaea detected in this study are discussed.

Extinction trends of threatened invertebrates in peninsular Spain

Using information from two recently published atlases of threatened invertebrate species in peninsular Spain, we examined the climatic, land use and geographic characteristics of the 100 km2 UTM cells with most likelihood of suffering extinctions (extinction cells), as well as the attributes of the species prone to population extinctions. Extinction cells have had significantly (1) lower precipitation values, (2) higher temperatures, (3) higher percentages of anthropic land uses or (4) higher rates of anthropization during the last 20 years than the remaining cells. Nevertheless, probable extinctions may occur under a wide range of climatic and anthropization change rates and these variables can only explain a low proportion (~5 %) of variability in the occurrence or number of extinction cells. Aquatic species seem to suffer higher local extinction rates than terrestrial species. Interestingly, many invertebrate species with approximately 25 or less occurrence cells are on a clear trajectory towards extinction. These results outline the difficulties and uncertainties in relating probable population extinctions with climatic and land use changes in the case of invertebrate data. However, they also suggest that a third of the considered Spanish threatened species could have lost some of their populations, and that current conservation efforts are insufficient to reverse this tendency.

Social and immigration factors in intimate partner violence among Ecuadorians, Moroccans and Romanians living in Spain

Background: Intimate partner violence (IPV) against women occurs in all countries, all cultures and at every level of society; however, some populations may be at greater risk than others. The aim of this study was to explore IPV prevalence among Ecuadorian, Moroccan and Romanian immigrant women living in Spain and its possible association with their personal, family, social support and immigration status characteristics. Methods: Cross-sectional study of 1607 adult immigrant women residing in Barcelona, Madrid and Valencia (2011). Prevalence rates and adjusted odds ratios (AORs) were calculated, with current IPV being the outcome. Different women’s personal (demographic), family, social support and immigration status characteristics were considered as explicative and control variables. All analyses were separated by women’s country of origin. Results: Current IPV prevalence was 15.57% in Ecuadorians, 10.91% in Moroccans and 8.58% in Romanians. Some common IPV factors were found, such as being separated and/or divorced. In Romanians, IPV was also associated with lack of social support [AOR 5.96 (1.39–25.62)] and low religious involvement [AOR 2.17 (1.06–4.43)]. The likelihood of current IPV was lower among women without children or other dependants in this subgroup [AOR 0.29 (0.093–0.92)]. Conclusion: The IPV prevalence rates obtained for Moroccan, Romanian and Ecuadorian women residing in Spain were similar. Whereas the likelihood of IPV appeared to be relatively evenly distributed among Moroccan and Ecuadorian women, it was higher among Romanian women in socially vulnerable situations related to family responsibilities and the lack of support networks. The importance of intervention in the process of separation and divorce was common to all women.

Professionals’ Perceptions of Support Resources for Battered Immigrant Women Chronicle of an Anticipated Failure

The aim of this study was to explore the experience of service providers in Spain regarding their daily professional encounters with battered immigrant women and their perception of this group’s help-seeking process and the eventual abandonment of the same. Twenty-nine in-depth interviews and four focus group discussions were conducted with a total of 43 professionals involved in providing support to battered immigrant women. We interviewed social workers, psychologists, intercultural mediators, judges, lawyers, and public health professionals from Spain. Through qualitative content analysis, four categories emerged: (a) frustration with the victim’s decision to abandon the help-seeking process, (b) ambivalent positions regarding differences between immigrant and Spanish women, (c) difficulties in the migratory process that may hinder the help-seeking process, and (d) criticisms regarding the inefficiency of existing resources. The four categories were cross-cut by an overarching theme: helping immigrant women not to abandon the help-seeking process as a chronicle of anticipated failure. The main reasons that emerged for abandoning the help-seeking process involved structural factors such as economic dependence, loss of social support after leaving their country of origin, and limited knowledge about available resources. The professionals perceived their encounters with battered immigrant women to be frustrating and unproductive because they felt that they had few resources to back them up. They felt that despite the existence of public policies targeting intimate partner violence (IPV) and immigration in Spain, the resources dedicated to tackling gender-based violence were insufficient to meet battered immigrant women’s needs. Professionals should be trained both in the problem of IPV and in providing support to the immigrant population.

Catheter-related bloodstream infection: burden of disease in a tertiary hospital

Background: Surveillance programmes have become the most effective tool for controlling catheter-related bloodstream infections (CRBSI). However, few studies have investigated programmes covering all hospital settings. Aim: To describe the results of a control and prevention programme for CRBSI based on compliance with recommendations for insertion and maintenance, using annual burden of disease in a tertiary level hospital. Methods: A CRBSI control and prevention programme involving all hospital settings was implemented. The programme consisted of CRBSI surveillance, direct observation of insertion and maintenance of catheters to determine performance, and education for healthcare workers. Findings: In total, 2043 short-term catheters were inserted in 1546 patients for 18,570 catheter-days, and 279 long-term catheters were inserted in 243 patients for 40,440 catheter-days. The annual incidence density was 5.98 (first semester 6.40, second semester 5.64) CRBSI per 1000 catheter-days for short-term catheters, and 0.57 (first semester 0.66, second semester 0.43) CRBSI per 1000 catheter-days for long-term catheters. One hundred and forty insertion procedures were observed, with an average insertion time of 13 (standard deviation 7) min. Compliance with recommendations was as follows: hand hygiene, 86.8%; use of alcoholic chlorhexidine solution for skin disinfection, 35.5%; use of mask, 93.4%; use of gloves, 98.7%; use of gown, 75.0%; use of sterile cloth, 93.8%; use of cap, 92.2%; bandage application, 62.7%; and use of aseptic technique, 89.5%. Forty-five maintenance procedures were observed, and compliance rates were as follows: hand hygiene, 42.1%; use of gloves, 78.1%; and port disinfection with alcoholic chlorhexidine solution, 32.5%. Conclusion: The CRBSI control and prevention programme implemented at the study hospital has decreased the rate of CRBSI, provided important information about the total burden of disease, and revealed possible ways to improve interventions in the future.

Brackish groundwater desalination by reverse osmosis in southeastern Spain. Presence of emerging contaminants and potential impacts on soil-aquifer media

Desalinated brackish groundwater is becoming a new source of water supply to comply with growing water demands, especially in (semi-) arid countries. Recent publications show that some chemical compounds may persist in an unaltered form after the desalination processes and that there is an associated risk of mixing waters with different salinity for irrigation. At the university of Alicante campus (Spain), a mix of desalinated brackish groundwater and water from the existing aquifer is currently applied for landscape irrigation. The presence of 209 emerging compounds, surfactants, priority substances according to the 2008/105/EC Directive, 11 heavy metals and microbiological organisms in blended water and aquifer samples was investigated. Thirty-five compounds were detected (pesticides, pharmaceuticals and surfactants) among them two priority substances α-endosulfan and Ni were found above the permitted maximum concentration. Blended water used for landscape irrigation during the summer period is supersaturated with respect to carbonates, which may ultimately lead to mineral precipitation in the soil-aquifer media and changes in hydraulic parameters.

Generation of nitrogen functionalities on activated carbons by amidation reactions and Hofmann rearrangement: Chemical and electrochemical characterization

Nitrogen functionalization of a highly microporous activated carbon (BET surface area higher than 3000 m2/g) has been achieved using the following sequence of treatments: (i) chemical oxidation using concentrated nitric acid, (ii) amidation by acyl chloride substitution with NH4NO3 and (iii) amination by Hoffman rearrangement. This reaction pathway yielded amide and amine functional groups, and a total nitrogen content higher than 3 at.%. It is achieved producing only a small decrease (20%) of the starting microporosity, being most of it related to the initial wet oxidation of the activated carbon. Remarkably, nitrogen aromatic rings were also formed as a consequence of secondary cyclation reactions. The controlled step-by-step modification of the surface chemistry allowed to assess the influence of individual nitrogen surface groups in the electrochemical performance in 1 M H2SO4 of the carbon materials. The largest gravimetric capacitance was registered for the pristine activated carbon due to its largest apparent surface area. The nitrogen-containing activated carbons showed the highest surface capacitances. Interestingly, the amidated activated carbon showed the superior capacitance retention due to the presence of functional groups (such as lactams, imides and pyrroles) that enhance electrical conductivity through their electron-donating properties, showing a capacitance of 83 F/g at 50 A/g.

Construction of 17-19th century domes in the province of Alicante (Spain): the case of San Juan Bautista in Cox

From the late seventeenth to early nineteenth centuries, many religious temples have been built in the province of Alicante (south east of Spain) with brick domes as their main characteristic feature. Often, the limited data available about these remarkable constructions make rehabilitation interventions become into real research projects, with a high value for their historic conservation over time. The aim of this paper is to show a detailed refurbishment analysis of a religious temple built in 1778, showing the need of preservation of historic buildings as a part of the architectural heritage by establishing a common pattern of materials, geometry and constructive systems, specifically in their domes. In most cases, there was not an architectural project for the construction, that is why the analysis of any documentary and archival sources available is essential to find different ways to proceed on the use and maintenance of these religious buildings.

Validation of the Spanish version of mackey childbirth satisfaction rating scale

Background: The “Mackey Childbirth Satisfaction Rating Scale” (MCSRS) is a complete non-validated scale which includes the most important factors associated with maternal satisfaction. Our primary purpose was to describe the internal structure of the scale and validate the reliability and validity of concept of its Spanish version MCSRS-E. Methods: The MCSRS was translated into Spanish, back-translated and adapted to the Spanish population. It was then administered following a pilot test with women who met the study participant requirements. The scale structure was obtained by performing an exploratory factorial analysis using a sample of 304 women. The structures obtained were tested by conducting a confirmatory factorial analysis using a sample of 159 women. To test the validity of concept, the structure factors were correlated with expectations prior to childbirth experiences. McDonald’s omegas were calculated for each model to establish the reliability of each factor. The study was carried out at four University Hospitals; Alicante, Elche, Torrevieja and Vinalopo Salud of Elche. The inclusion criteria were women aged 18–45 years old who had just delivered a singleton live baby at 38–42 weeks through vaginal delivery. Women who had difficulty speaking and understanding Spanish were excluded. Results: The process generated 5 different possible internal structures in a nested model more consistent with the theory than other internal structures of the MCSRS applied hitherto. All of them had good levels of validation and reliability. Conclusions: This nested model to explain internal structure of MCSRS-E can accommodate different clinical practice scenarios better than the other structures applied to date, and it is a flexible tool which can be used to identify the aspects that should be changed to improve maternal satisfaction and hence maternal health.

High Speed Rail Passenger Profile in Sun and Beach Tourism Destinations: The Case of Alicante (Spain)

This paper aims to study and highlight the profile of the high speed rail (HSR) passenger in the case of the well-known sun and beach destination of Alicante, located in the Southeast of Spain. This region, which is directly connected with Madrid, differs from others because of its warm and sunny weather. The province is a tourist destination for both Spanish citizens and foreigners. However, the studies on the dynamics of the corridor towards Madrid and Alicante regarding HSR are quite recent and it is not possible to draw final conclusions about its impact on tourist mobility.