Highly exposed segment of the Spf1p P5AATPase near transmembrane M5 detected by limited proteolysis

The yeast Spf1p protein is a primary transporter that belongs to group 5 of the large family of P-ATPases. Loss of Spf1p function produces ER stress with alterations of metal ion and sterol homeostasis and protein folding, glycosylation and membrane insertion. The amino acid sequence of Spf1p shows the characteristic P-ATPase domains A, N, and P and the transmembrane segments M1-M10. In addition, Spf1p exhibits unique structures at its N-terminus (N-T region), including two putative additional transmembrane domains, and a large insertion connecting the P domain with transmembrane segment M5 (D region). Here we used limited proteolysis to examine the structure of Spf1p. A short exposure of Spf1p to trypsin or proteinase K resulted in the cleavage at the N and C terminal regions of the protein and abrogated the formation of the catalytic phosphoenzyme and the ATPase activity. In contrast, limited proteolysis of Spf1p with chymotrypsin generated a large N-terminal fragment containing most of the M4-M5 cytosolic loop, and a minor fragment containing the C-terminal region. If lipids were present during chymotryptic proteolysis, phosphoenzyme formation and ATPase activity were preserved. ATP slowed Spf1p proteolysis without detectable changes of the generated fragments. The analysis of the proteolytic peptides by mass spectrometry and Edman degradation indicated that the preferential chymotryptic site was localized near the cytosolic end of M5. The susceptibility to proteolysis suggests an unexpected exposure of this region of Spf1p that may be an intrinsic feature of P5A-ATPases.Fil: Petrovich, Guido Daniel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Corradi, Gerardo Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; ArgentinaFil: Pavan, Carlos Humberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; ArgentinaFil: Noli Truant, Sofia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Adamo, Hugo Pedro. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentin

The concept and measurement of violence against women and men

Available Open Access under CC-BY-NC licence. The extent of violence against women is currently hidden. How should violence be measured? How should research and new ways of thinking about violence improve its measurement? Could improved measurement change policy? The book is a guide to how the measurement of violence can be best achieved. It shows how to make femicide, rape, domestic violence, and FGM visible in official statistics. It offers practical guidance on definitions, indicators and coordination mechanisms. It reflects on theoretical debates on ‘what is gender’, ‘what is violence’, and ‘the concept of coercive control’. and introduces the concept of ‘gender saturated context’. Analysing the socially constructed nature of statistics and the links between knowledge and power, it sets new standards and guidelines to influence the measurement of violence in the coming decades

Loss of choline agonism in the inner ear hair cell nicotinic acetylcholine receptor linked to the alpha10 subunit

The α9α10 nicotinic acetylcholine receptor (nAChR) plays a fundamental role in inner ear physiology. It mediates synaptic transmission between efferent olivocochlear fibers that descend from the brainstem and hair cells of the auditory sensory epithelium. The α9 and α10 subunits have undergone a distinct evolutionary history within the family of nAChRs. Predominantly in mammalian vertebrates, the α9α10 receptor has accumulated changes at the protein level that may ultimately relate to the evolutionary history of the mammalian hearing organ. In the present work, we investigated the responses of α9α10 nAChRs to choline, the metabolite of acetylcholine degradation at the synaptic cleft. Whereas choline is a full agonist of chicken α9α10 receptors it is a partial agonist of the rat receptor. Making use of the expression of α9α10 heterologous receptors, encompassing wild-type, heteromeric, homomeric, mutant, chimeric, and hybrid receptors, and in silico molecular docking, we establish that the mammalian (rat) α10 nAChR subunit underscores the reduced efficacy of choline. Moreover, we show that whereas the complementary face of the α10 subunit does not play an important role in the activation of the receptor by ACh, it is strictly required for choline responses. Thus, we propose that the evolutionary changes acquired in the mammalian α9α10 nAChR resulted in the loss of choline acting as a full agonist at the efferent synapse, without affecting the triggering of ACh responses. This may have accompanied the fine-tuning of hair cell post-synaptic responses to the high-frequency activity of efferent medial olivocochlear fibers that modulate the cochlear amplifier.Fil: Moglie, Marcelo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Marcovich, Irina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Corradi, Jeremias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Carpaneto Freixas, Agustín Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Gallino, Sofia Ludmila. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Plazas, Paola Viviana. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Farmacologia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Lipovsek, Maria Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Elgoyhen, Ana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentin

Multi-wavelength observations of a nearby multi-phase interstellar cloud

High-resolution spectroscopic observations (UV HST/STIS and optical) are used to characterize the physical state and velocity structure of the multiphase interstellar medium seen towards the nearby (170 pc) star HD102065, located behind the tail of a cometary-shaped, infrared cirrus-cloud, in the area of interaction between the Sco-Cen OB association and the Local Bubble. We analyze interstellar components present along the line of sight by fitting multiple transitions from CO, CH, CH+, C I, S I, Fe I, Mg I, Mg II, Mn II, P II, Ni II, C II, N I, O I, Si III, C IV, and Si IV. The absorption spectra are complemented by H I, CO and C II emission-line spectra, H$_2$ column-densities derived from FUSE spectra, and IRAS images. Gas components of a wide range of temperatures and ionization states are detected along the line of sight. Most of the hydrogen column-density is in cold, diffuse, molecular gas at low LSR velocity. This gas is mixed with traces of warmer molecular gas traced by H2 in the J>2 levels, in which the observed CH+ must be formed. We also identify three distinct components of warm gas at negative velocities down to -20 km/s. The temperature and gas excitation are shown to increase with increasing velocity shift from the bulk of the gas. Hot gas at temperatures of several 10^5 K is detected in the most negative velocity component in the highly-ionized species. This hot gas is also detected in very strong lines of less-ionized species (Mg II, Si II* and C II*) for which the bulk of the gas is cooler. We relate the observational results to evidence for dynamical impact of the Sco-Cen stellar association on the nearby ISM. We propose a scenario where the cirrus cloud has been hit a few 10^5 yr ago by a supernova blast wave originating from the Lower Centaurus Crux group of the Sco-Cen association.Comment: 16 pages, 9 figures, to be published in A&

Interstellar extinction and polarization -- A spheroidal dust grain approach perspective

We extend and investigate the spheroidal model of interstellar dust grains used to simultaneously interpret the observed interstellar extinction and polarization curves. We compare our model with similar models recently suggested by other authors, study its properties and apply it to fit the normalized extinction $A(\lambda)/A_{\rm V}$ and the polarizing efficiency $P(\lambda)/A(\lambda)$ measured in the near IR to far UV region for several stars seen through one large cloud. We conclude that the model parameter $\Omega$ being the angle between the line of sight and the magnetic field direction can be more or less reliably determined from comparison of the theory and observations. This opens a way to study the spatial structure of interstellar magnetic fields by using multi-wavelength photometric and polarimetric observations.Comment: 11 pages, 4 figures and 4 tables, To appear in MNRAS (added

Lights4Violence: a quasi-experimental educational intervention in six European countries to promote positive relationships among adolescents.

Background Preventing intimate partner violence or dating violence (DV) among adolescents is a public health priority due to its magnitude and damaging short and long-term consequences for adolescent and adult health. In our study protocol, we complement prior experiences in DV prevention by promoting protective factors (or assets) against gender violence such as communication skills, empathy and problem-solving capability through “Cinema Voice”, a participatory educational intervention based on adolescents’ strengths to tackle DV. Methods/design A longitudinal quasi-experimental educational intervention addressed to boys and girls ages 13–17 years, enrolled in secondary education schools in Alicante (Spain), Rome (Italy), Cardiff (UK), Iasi (Romania), Poznan (Poland) and Matosinhos (Portugal). Both process and results evaluations will be carried out with 100–120 intervention and 120–150 control group students per city at three time periods: before, after and 6 months after the implementation of the following interventions: 1) Training seminar with teachers to promote knowledge and skills on the core issues of intervention; 2) Workshops with intervention groups, where participants produce their own digital content presenting their perspective on DV; and 3) Short film exhibitions with participants, their families, authorities and other stakeholders with the objective of share the results and engage the community. Outcome measures are self-perceived social support, machismo, sexism, tolerance towards gender violence, social problem-solving and assertiveness as well as involvement in bullying/cyberbullying. Other socio-demographic, attitudes and violence-related co-variables were also included. Discussion This study may provide relevant information about the effectiveness of educational interventions that combine a positive youth development framework with educational awareness about the importance of achieving gender equality and preventing and combating gender violence. To our knowledge, this is the first study that involves six European countries in an educational intervention to promote violence protective assets among enrolled adolescents in secondary schools. This study may provide the needed tools to replicate the experience in other contexts and other countrie

The concept and measurement of violence against women and men

Available Open Access under CC-BY-NC licence. The extent of violence against women is currently hidden. How should violence be measured? How should research and new ways of thinking about violence improve its measurement? Could improved measurement change policy? The book is a guide to how the measurement of violence can be best achieved. It shows how to make femicide, rape, domestic violence, and FGM visible in official statistics. It offers practical guidance on definitions, indicators and coordination mechanisms. It reflects on theoretical debates on ‘what is gender’, ‘what is violence’, and ‘the concept of coercive control’. and introduces the concept of ‘gender saturated context’. Analysing the socially constructed nature of statistics and the links between knowledge and power, it sets new standards and guidelines to influence the measurement of violence in the coming decades

Non-invAsive VentIlation for early General wArd respiraTory failurE (NAVIGATE):A multicenter randomized controlled study. Protocol and statistical analysis plan

Few randomized trials have evaluated the use of non-invasive ventilation (NIV) for early acute respiratory failure (ARF) in non-intensive care unit (ICU) wards. The aim of this study is to test the hypothesis that early NIV for mild-moderate ARF in non-ICU wards can prevent development of severe ARF

Exhaled and arterial levels of endothelin-1 are increased and correlate with pulmonary systolic pressure in COPD with pulmonary hypertension

BACKGROUND: Endothelin-1 (ET-1) and Nitric Oxide (NO) are crucial mediators for establishing pulmonary artery hypertension (PAH). We tested the hypothesis that their imbalance might also occur in COPD patients with PAH. METHODS: The aims of the study were to measure exhaled breath condensate (EBC) and circulating levels of ET-1, as well as exhaled NO (FENO) levels by, respectively, a specific enzyme immunoassay kit, and by chemiluminescence analysis in 3 groups of subjects: COPD with PAH (12), COPD only (36), and healthy individuals (15). In order to evaluate pulmonary-artery systolic pressure (PaPs), all COPD patients underwent Echo-Doppler assessment. RESULTS: Significantly increased exhaled and circulating levels of ET-1 were found in COPD with PAH compared to both COPD (p < 0.0001) only, and healthy controls (p < 0.0001). In COPD with PAH, linear regression analysis showed good correlation between ET-1 in EBC and PaPs (r = 0.621; p = 0.031), and between arterial levels of ET-1 and PaPs (r = 0.648; p = 0.022), while arterial levels of ET-1 inversely correlated with FEV1%, (r = -0.59, p = 0.043), and PaPs negatively correlated to PaO2 (r = -0.618; p = 0.032). Significantly reduced levels of FENO were found in COPD associated with PAH, compared to COPD only (22.92 +/- 11.38 vs.35.07 +/- 17.53 ppb; p = 0.03). Thus, we observed an imbalanced output in the breath between ET-1 and NO, as expression of pulmonary endothelium and epithelium impairment, in COPD with PAH compared to COPD only. Whether this imbalance is an early cause or result of PAH due to COPD is still unknown and deserves further investigations