P2X1 and P2X5 subunits form the functional P2X receptor in mouse cortical astrocytes

ATP plays an important role in signal transduction between neuronal and glial circuits and within glial networks. Here we describe currents activated by ATP in astrocytes acutely isolated from cortical brain slices by non-enzymatic mechanical dissociation. Brain slices were prepared from transgenic mice that express enhanced green fluorescent protein under the control of the human glial fibrillary acidic protein promoter. Astrocytes were studied by whole-cell voltage clamp. Exogenous ATP evoked inward currents in 75 of 81 astrocytes. In the majority (~65%) of cells, ATP-induced responses comprising a fast and delayed component; in the remaining subpopulation of astrocytes, ATP triggered a smoother response with rapid peak and slowly decaying plateau phase. The fast component of the response was sensitive to low concentrations of ATP (with EC50 of ~40 nM). All ATP-induced currents were blocked by pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS); they were insensitive to ivermectin. Quantitative real-time PCR demonstrated strong expression of P2X1 and P2X5 receptor subunits and some expression of P2X2 subunit mRNAs. The main properties of the ATP-induced response in cortical astrocytes (high sensitivity to ATP, biphasic kinetics, and sensitivity to PPADS) were very similar to those reported for P2X1/5 heteromeric receptors studied previously in heterologous expression systems

Bayes Turchin analysis of X ray absorption data above the Fe L2,3 edges

Extended X ray absorption fine structure EXAFS data and magnetic EXAFS MEXAFS data were measured at two temperatures 180 K and 296 K in the energy region of the overlapping L edges of bcc Fe grown on a V 110 crystal surface. In combination with a Bayes Turchin data analysis procedure these measurements enable the exploration of local crystallographic and magnetic structures. The analysis determined the atomic like background together with the EXAFS parameters which consisted of 10 shell radii, the Debye Waller parameters, separated into structural and vibrational components, and the third cumulant of the first scattering path. The vibrational components for 97 different scattering paths were determined by a two parameter force field model using a priori values adjusted to Born von Karman parameters of inelastic neutron scattering data. The investigations of the system Fe V 110 demonstrate that the simultaneous fitting of atomic background parameters and EXAFS parameters can be performed reliably. Using the L2 and L3 components extracted from the EXAFS analysis and the rigid band model, the MEXAFS oscillations can only be described when the sign of the exchange energy is changed compared to the predictions of the Hedin Lundquist exchange and correlation functiona

Theoretical X-Ray Absorption Debye-Waller Factors

An approach is presented for theoretical calculations of the Debye-Waller factors in x-ray absorption spectra. These factors are represented in terms of the cumulant expansion up to third order. They account respectively for the net thermal expansion $\sigma^{(1)}(T)$, the mean-square relative displacements $\sigma^2(T)$, and the asymmetry of the pair distribution function $\sigma^{(3)}(T)$. Similarly, we obtain Debye-Waller factors for x-ray and neutron scattering in terms of the mean-square vibrational amplitudes $u^2(T)$. Our method is based on density functional theory calculations of the dynamical matrix, together with an efficient Lanczos algorithm for projected phonon spectra within the quasi-harmonic approximation. Due to anharmonicity in the interatomic forces, the results are highly sensitive to variations in the equilibrium lattice constants, and hence to the choice of exchange-correlation potential. In order to treat this sensitivity, we introduce two prescriptions: one based on the local density approximation, and a second based on a modified generalized gradient approximation. Illustrative results for the leading cumulants are presented for several materials and compared with experiment and with correlated Einstein and Debye models. We also obtain Born-von Karman parameters and corrections due to perpendicular vibrations.Comment: 11 pages, 8 figure

Bienestar animal aplicado a la producción bovina

La preocupación acerca del bienestar animal ha ido en aumento, particularmente por la presión del público consumidor. Se entiende por bienestar al estado del individuo en relación a sus intentos de adaptarse al ambiente en un momento determinado. Al ser un concepto con bases científicas debe ser medido objetivamente. Por otra parte, la definición tiene algunas implicancias como la de ser una característica del animal, en lugar de algo que se le pueda brindar, así como variar dentro de un rango –desde muy bueno a muy malo– siendo malo cuando existen dificultades para mantener el control de las funciones o fallas en las tentativas de adaptarse al ambiente. En la cría y producción de bovinos existe una opinión generalizada que los sistemas extensivos son siempre buenos, porque mantienen a los animales lo más cerca posible del estado natural y que los sistemas intensivos son malos porque las libertades de elección y movimientos están restringidas. No obstante, en cualquier sistema productivo, extensivo o intensivo, pueden existir fracasos en la adaptación y consecuente estrés. Actualmente algunas organizaciones internacionales se han planteado como objetivo lograr que las nuevas tecnologías mejoren el bienestar sin producir mermas en la producción. Esto se puede alcanzar mediante el reconocimiento de los comportamientos naturales del bovino, teniendo en cuenta sus señales visuales, auditivas y olfativas, bases del manejo racional de los animales. Esta práctica no solo permite mejorar la relación humano–animal disminuyendo los signos de estrés, sino que también proporciona beneficios prácticos y económicos, generando efectos psicosociales positivos para el bienestar humano.

Problemas de agresividad canina. Estudio preliminar en las ciudades de Corrientes y Resistencia

La agresión canina es un problema de comportamiento que supone un riesgo potencial para la sociedad. En este estudio se evaluó el comportamiento de 120 perros mediante el “Cuestionario de Evaluación e Investigación del Comportamiento de los Caninos” (C-BARQ) con el objeto de evidenciar la presencia de conductas agresivas en perros cuyos propietarios acudieron a la consulta clínica privada por causas ajenas a este problema, en las ciudades de Corrientes y Resistencia (Argentina). Del análisis descriptivo de los datos se pudo apreciar que un 55% de los perros mostró agresión hacia los propietarios. Si bien la frecuencia promedio no fue alta, la presencia de conducta agresiva implica un riesgo potencial para las personas. Además, se debe tener en cuenta que ninguno de los propietarios de los canes acudió a la clínica veterinaria para consultar por un problema asociado al comportamiento. Se observa que los dueños de los perros no consideran importantes a las señales sutiles de agresión, ignorando el peligro que podrían significar. Ello impone la necesidad de continuar realizando estudios sobre el tema y de analizar una forma efectiva de educación e información

Prescission neutron multiplicity and fission probability from Langevin dynamics of nuclear fission

A theoretical model of one-body nuclear friction which was developed earlier, namely the chaos-weighted wall formula, is applied to a dynamical description of compound nuclear decay in the framework of the Langevin equation coupled with statistical evaporation of light particles and photons. We have used both the usual wall formula friction and its chaos-weighted version in the Langevin equation to calculate the fission probability and prescission neutron multiplicity for the compound nuclei $^{178}$W, $^{188}$Pt, $^{200}$Pb, $^{213}$Fr, $^{224}$Th, and $^{251}$Es. We have also obtained the contributions of the presaddle and postsaddle neutrons to the total prescission multiplicity. A detailed analysis of our results leads us to conclude that the chaos-weighted wall formula friction can adequately describe the fission dynamics in the presaddle region. This friction, however, turns out to be too weak to describe the postsaddle dynamics properly. This points to the need for a suitable explanation for the enhanced neutron emission in the postsaddle stage of nuclear fission.Comment: RevTex, 14 pages including 5 Postscript figures, results improved by using a different potential, conclusions remain unchanged, to appear in Phys. Rev.

Weight management: a comparison of existing dietary approaches in a work-site setting

<b>OBJECTIVES:</b> (1) To compare the effectiveness a 2512 kJ (600 kcal) daily energy deficit diet (ED) with a 6279 kJ (1500 kcal) generalized low-calorie diet (GLC) over a 24 week period (12 weeks weight loss plus 12 weeks weight maintenance). (2) To determine if the inclusion of lean red meat at least five times per week as part of a slimming diet is compatible with weight loss in comparison with a diet that excludes lean red meat. DESIGN: Randomized controlled trial. <b>SETTING:</b> Large petrochemical work-site. <b>PARTICIPANTS:</b> One-hundred and twenty-two men aged between 18 and 55 y. <b>MAIN OUTCOME MEASURES:</b> Weight loss and maintenance of weight loss. <b>INTERVENTION:</b> Eligible volunteers were randomized to one of the four diet=meat combinations (ED meat, ED no meat, GLC meat, GLC no meat). One-third of subjects in each diet/meat combination were randomized to an initial control period prior to receiving dietary advice. All subjects attended for review every 2 weeks during the weight loss period. For the 12 week structured weight maintenance phase, individualized energy prescriptions were re-calculated for the ED group as 1.4 (activity factor)x basal metabolic rate. Healthy eating advice was reviewed with subjects in the GLC group. All subjects were contacted by electronic mail at 2 week intervals and anthropometric and dietary information requested. <b>RESULTS:</b> No difference was evident between diet groups in mean weight loss at 12 weeks (4.3 (s.d. 3.4) kg ED group vs 5.0 (s.d. 3.5) kg GLC group, P=0.34). Mean weight loss was closer to the intended weight loss in the 2512 kJ (600 kcal) ED group. The dropout rate was also lower than the GLC group. The inclusion of lean red meat in the diet on at least five occasions per week did not impair weight loss. Mean weight gain following 12 weeks weight maintenance was þ1.1 (s.d. 1.8) kg, P<0.0001. No differences were found between groups. <b>CONCLUSIONS:</b> This study has shown that the individualized 2512 kJ (600 kcal) ED approach was no more effective in terms of weight loss than the 6279 kJ (1500 kcal) GLC approach. However the ED approach might be considered preferable as compliance was better with this less demanding prescription. In terms of weight loss the elimination of red meat from the diet is unnecessary. The weight maintenance intervention was designed as a low-input approach, however weight regain was significant and weight maintenance strategies require further development

Biomarkers of exposure and effect—interpretation in human risk assessment

The effect of exposure to carcinogenic polycyclic aromatic hydrocarbons adsorbed onto respirable air particles (PM2.5, diameter < 2.5 μm) on DNA adducts and chromosomal aberrations was repeatedly studied in Prague, Czech Republic, in groups of policemen working in the downtown area and in bus drivers. Personal exposure was evaluated using personal samplers during working shifts. DNA adducts were analyzed in lymphocytes by the 32P-postlabeling assay and chromosomal aberrations were analyzed by conventional cytogenetic analysis and fluorescent in situ hybridization (FISH). The impact of environmental pollution on DNA adducts and chromosomal aberrations was studied in a total of 950 subjects. Our results suggest that the environmental exposure of nonsmokers to concentrations higher than 1 ng benzo[a]pyrene/m3 represents a risk of DNA damage, as indicated by an increase in DNA adducts and the genomic frequency of translocations determined by FISH

Multiplexed profiling of GPCR activities by combining split TEV assays and EXT-based barcoded readouts

G protein-coupled receptors (GPCRs) are the largest class of cell surface receptors and are implicated in the physiological regulation of many biological processes. The high diversity of GPCRs and their physiological functions make them primary targets for therapeutic drugs. For the generation of novel compounds, however, selectivity towards a given target is a critical issue in drug development as structural similarities between members of GPCR subfamilies exist. Therefore, the activities of multiple GPCRs that are both closely and distantly related to assess compound selectivity need to be tested simultaneously. Here, we present a cell-based multiplexed GPCR activity assay, termed GPCRprofiler, which uses a beta-arrestin recruitment strategy and combines split TEV protein-protein interaction and EXT-based barcode technologies. This approach enables simultaneous measurements of receptor activities of multiple GPCR-ligand combinations by applying massively parallelized reporter assays. In proof-of-principle experiments covering 19 different GPCRs, both the specificity of endogenous agonists and the polypharmacological effects of two known antipsychotics on GPCR activities were demonstrated. Technically, normalization of barcode reporters across individual assays allows quantitative pharmacological assays in a parallelized manner. In summary, the GPCRprofiler technique constitutes a flexible and scalable approach, which enables simultaneous profiling of compound actions on multiple receptor activities in living cells

Alteraciones hematológicas en perros (Canis lupus familiaris) diagnosticados con Ehrlichia spp. por PCR, en clínicas veterinarias del Nordeste Argentino

Ehrlichiosis is a vector-borne disease that affects numerous species. It is caused by Ehrlichia spp. gram-negative, obligate intracellular bacteria with a coccoid or pleomorphic appearance that infects monocytes. Diagnosis, through clinical symptoms, is difficult given the similarity of presentation with other vector-borne disease. The evaluation of hematological parameters (hemogram and blood smear) is the first indication of the clinical veterinarian, since it is possible to find intracytoplasmic inclusions (morulae) in monocytes. The aim of this study was to evaluate hematological alterations in dogs diagnosed with Ehrlichia spp. by peripheral blood smear, and to confirm it by molecular technique (PCR). From June/2021 to July/2022 dog samples were analyzed and intracytoplasmic inclusions were identified in seven samples. The presence of anemia and thrombocytopenia without alterations in the number of leukocytes was observed in 6 of 7 blood samples. The PCR technique confirmed the initial diagnosis in 2/7 samples, amplifying the dsb 330/728 gene segment. The usefulness of hematological parameters together with the diagnostic confirmation by PCR technique is emphasized, the latter being a highly sensitive and specific method, which offers up to 100% diagnostic confidence in detecting and amplifying the DNA of Ehrlichia spp.La ehrlichiosis es una enfermedad transmitida por vectores (ETV) que afecta a numerosas especies. Es causada por Ehrlichia spp. una bacteria gramnegativa, intracelular obligada, de aspecto cocoide o pleomórfica, que infecta los monocitos. El diagnóstico, a través de los síntomas clínicos, se dificulta dada la similitud de presentación con otras ETV. La evaluación de los parámetros hematológicos (hemograma y extendido sanguíneo) es la primera indicación del veterinario clínico, ya que es posible encontrar inclusiones intracitoplasmáticas (mórulas) en los monocitos. El objetivo del trabajo fue evaluar las alteraciones hematológicas de perros diagnosticados con Ehrlichia spp. por extendido de sangre periférica, y confirmarlo por técnica molecular (PCR). De junio/2021 a julio/2022 se analizaron muestras de perros identificando inclusiones intracitoplasmáticas en siete muestras. Se observó en 6 de 7 muestras de sangre la presencia de anemia y trombocitopenia sin alteraciones del número de leucocitos. La técnica de PCR confirmo el diagnóstico inicial en 2/7 muestras, amplificando el segmento del gen dsb 330/728. Se destaca la utilidad de los parámetros hematológicos junto a la confirmación diagnóstica por la técnica de PCR, siendo esta última, un método altamente sensible y específico, que ofrece hasta un 100% de seguridad en el diagnóstico detectando y amplificando el ADN de Ehrlichia spp