Sistematización de la metodología de validación e interpretación de resultados para medidas clínicas de la enfermedad de Parkinson

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Medicina Preventiva y Salud Pública y Microbiología. Fecha de lectura: 12-03-201

Biosafety steps in the manufacturing process of spray-dried plasma: a review with emphasis on the use of ultraviolet irradiation as a redundant biosafety procedure

Spray dried plasma (SDP) is a functional protein source obtained from blood of healthy animals, approved by the veterinary authorities from animals declared to be fit for slaughter for human consumption. Blood of these animals is collected at the slaughterhouse, treated with an anticoagulant, chilled and transported to industrial facilities in which blood is centrifuged to separate the red blood cells from the plasma fraction. Plasma is then concentrated, and spray dried at high temperatures (80 °C throughout its substance) to convert it in a powder. Such method preserves the biological activity of its proteins, mainly albumins and globulins. SDP is mainly used in pig feed diets to significantly improve daily gain, feed intake, production efficiency, and to reduce post-weaning lag caused by the appearance of post-weaning diarrhea. Although SDP is considered a safe product and its manufacturing process consists of several biosafety steps, the security of the SDP is often questioned due to its nature as raw blood by-product, especially when emergent or re-emergent pathogens appear. This review provides an evaluation and validation of the different safety steps present in the manufacturing process of SDP, with special focus on a new redundant pathogen inactivation step, the UV-C irradiation, that may be implemented in the manufacturing process of the SDP. Overall results showed that the manufacturing process of SDP is safe and the UV-C radiation was effective in inactivating a wide range of bacteria and viruses spiked and naturally present in commercially collected liquid animal plasma and it can be implemented as a redundant biosafety step in the manufacturing process of the SDP.info:eu-repo/semantics/publishedVersio

Commercial feed containing porcine plasma spiked with African swine fever virus is not infective in pigs when administered for 14 consecutive days

The objective of this study was to determine if commercially collected liquid porcine plasma contaminated with African swine fever virus (ASFV) and fed for 14 consecutive days would infect pigs. Commercially collected liquid porcine plasma was mixed with the serum from an ASFV experimentally infected pig. To simulate the potential of pigs slaughtered being ASFV viremic but asymptomatic and passing antemortem inspection, the ratio of liquid plasma from healthy animals to serum from an ASFV infected pig used in this study represented 0.4% or 2.0% of the pigs slaughtered being viremic (Studies 1 or 2, respectively). The contaminated liquid plasma was mixed on commercial feed and pigs were fed for 14 consecutive days providing to each pig 104.3 or 105.0 TCID50 ASFV daily (Studies 1 or 2, respectively). Pigs were observed for an additional 5 or 9 days (Studies 1 or 2, respectively). In both experiments, the pigs did not become infected with ASFV during the 14d feeding period or during the subsequent observation period. In these experiments, unprocessed liquid plasma contaminated with ASFV mixed on commercial feed and fed for 14 consecutive days did not infect pigs. From our results we can conclude that the infectious dose of ASFV on feed is much higher than that previously reported, at least with ASFV-spiked raw plasma.info:eu-repo/semantics/publishedVersio

Biosafety steps in the manufacturing process of spray-dried plasma : a review with emphasis on the use of ultraviolet irradiation as a redundant biosafety procedure

Ajuts: Secretaria de Universitats i Recerca del Departament d'Economia i Coneixement de la Generalitat de Catalunya 014 DI 066Spray dried plasma (SDP) is a functional protein source obtained from blood of healthy animals, approved by the veterinary authorities from animals declared to be fit for slaughter for human consumption. Blood of these animals is collected at the slaughterhouse, treated with an anticoagulant, chilled and transported to industrial facilities in which blood is centrifuged to separate the red blood cells from the plasma fraction. Plasma is then concentrated, and spray dried at high temperatures (80 °C throughout its substance) to convert it in a powder. Such method preserves the biological activity of its proteins, mainly albumins and globulins. SDP is mainly used in pig feed diets to significantly improve daily gain, feed intake, production efficiency, and to reduce post-weaning lag caused by the appearance of post-weaning diarrhea. Although SDP is considered a safe product and its manufacturing process consists of several biosafety steps, the security of the SDP is often questioned due to its nature as raw blood by-product, especially when emergent or re-emergent pathogens appear. This review provides an evaluation and validation of the different safety steps present in the manufacturing process of SDP, with special focus on a new redundant pathogen inactivation step, the UV-C irradiation, that may be implemented in the manufacturing process of the SDP. Overall results showed that the manufacturing process of SDP is safe and the UV-C radiation was effective in inactivating a wide range of bacteria and viruses spiked and naturally present in commercially collected liquid animal plasma and it can be implemented as a redundant biosafety step in the manufacturing process of the SDP

Activated char from the co-pyrolysis of polystyrene and olive stone mixtures for the adsorption of CO2

Yogurt plastic containers made of polystyrene (PS), olive stone, and mixtures of both have been converted into activated carbon materials transforming them firstly into char via pyrolysis and secondly with activation using either KOH or H2SO4. The pyrolysis of the olive stone gave a higher yield of material than the plastic PS. However, the activation of the PS char with KOH was more effective, reaching surface areas of 508 vs 194 m2 g−1 of the corresponding prepared with olive stone. The prepared materials were tested as CO2 adsorbent in thermobalance and fixed-be column assays. The materials activated with H2SO4 slightly enhanced the adsorption ability of the original char but were far from the performance obtained with KOH activation. The CO2 isotherms showed high synergy of CO2 uptake and selectivity when using activated chars prepared with the char from the mixture of raw materials, specially at a 1:1 mass ratio. The isosteric heat of adsorption values were those expected for a physisorption process. Further experiments in a fixed-bed column were also studied at atmospheric pressure at different inlet CO2 concentrations (10–50%). The CO2 retention increased as the partial CO2 pressure rose. Besides, a very similar performance of the material prepared with plastic and olive stones was obtained at 50%, i.e. 220 and 197 mg g−1 respectively. At low CO2 concentrations, the materials enriched with plastic displayed better performance than those prepared with olive stone. Cycles of adsorption-desorption were carried out in the column to assess the stability of the materials. The curves obtained did not display any substantial change, demonstrating the lack of adsorption retention

Estimated quantity of swine virus genomes based on quantitative PCR analysis in spray-dried porcine plasma samples collected from multiple manufacturing plants

This survey was conducted to estimate the incidence and level of potential viral contamination in commercially collected porcine plasma. Samples of spray dried porcine plasma (SDPP) were collected over a 12- month period from eight spray drying facilities in Spain, England, Northern Ireland, Brazil, Canada, and the United States. In this survey, viral load for several porcine pathogens including SVA, TGEV, PRRSV (EU and US strains), PEDV, PCV-2, SIV, SDCoV and PPV were determined by qPCR. Regression of Ct on TCID of serial diluted stock solution of each virus allowed the estimate of potential viral level in SDPP and unprocessed liquid plasma (using typical solids content of commercially collected porcine plasma). In this survey SVA, TGEV or SDCoV were not detected in any of the SDPP samples. Brazil SDPP samples were free of PRRSV and PEDV. Samples of SDPP from North America primarily contained the PRRSV-US strain while the European samples contained the PRRSV-EU strain (except for one sample from each region containing a relatively low estimated level of the alternative PRRSV strain). Estimated viral level tended to be in the range from <1.0 log TCID to <2.5 log TCID. Estimated level of SIV was the exception with a very low incidence rate but higher estimated viral load <3.9 log TCID. In summary, the incidence of potential viral contamination in commercially collected porcine plasma was variable and estimated virus level in samples containing viral DNA/RNA was relatively low compared with that occurring at the peak viremia during an infection for all viruses or when considering the minimal infectious dose for each of them

Evaluation of the effectiveness of the SurePure Turbulator ultraviolet-C irradiation equipment on inactivation of different enveloped and non-enveloped viruses inoculated in commercially collected liquid animal plasma

The objective of this study was to evaluate the effectiveness of the SurePure Turbulator ultraviolet-C (UV-C, 254 nm wavelength) irradiation equipment on inactivation of different enveloped and non-enveloped viruses in commercially collected liquid animal plasma. Specifically, Pseudorabies virus (PRV), Porcine reproductive and respiratory syndrome virus (PRRSV), Porcine epidemic diarrhea virus (PEDV), Bovine viral diarrhea virus (BVDV), Classical swine fever virus (CSFV), Swine influenza virus (SIV) as enveloped viruses and Porcine parvovirus (PPV), Swine vesicular disease virus (SVDV), Porcine circovirus type 2 (PCV-2) and Senecavirus A (SVA) as non-enveloped viruses, were inoculated in bovine or porcine plasma and subjected to different UV-C irradiation doses (0, 750, 1500, 3000, 6000 and 9000 J/L) using an UV-C device developed for opaque liquid working under turbulent flow. The enveloped viruses tested were inactivated at < 3000 J/L of UV-C, being the dose needed to inactivate 4 log TCID50 (4D) of 1612 J/L for PRV,1004 J/L for PRRSV, 1953 J/L for PEDV, 1639 J/L for SIV, 1641 J/L for CSFV and 1943 J/L for BVDV. The non-enveloped viruses tended to have higher 4D values: 2161 J/L for PPV, 3223 J/L for SVA and 3708 J/L for SVDV. Because the initial viral concentration was <4.0 Log for PCV-2, it was not possible to calculate the 4D value for this virus. In conclusion, these results demonstrated that the SurePure Turbulator UV-C treatment system is capable of inactivating significant levels of swine viruses inoculated in commercially collected porcine or bovine plasma. It was concluded that irradiation with UV-C can provide an additional redundant biosafety feature in the manufacturing process of spray-dried animal plasmainfo:eu-repo/semantics/publishedVersio

Effect of the Lights4Violence intervention on the sexism of adolescents in European countries

Background: Sexism results in a number of attitudes and behaviors that contribute to gender inequalities in social structure and interpersonal relationships. The objective of this study was to evaluate the effectiveness of Lights4Violence, an intervention program based on promoting health assets to reduce sexist attitudes in young European people. Methods: We carried out a quasi-experimental study in a non-probabilistic population of 1146 students, aged 12–17 years. The dependent variables were the difference in the wave 1 and wave 2 values in the subscales of the Ambivalent Sexism Inventory: benevolent sexism (BS) and hostile sexism (HS). The effect of the intervention was evaluated through linear regression analyses stratified by sex. The models were adjusted by baseline subscales scores, socio-demographic and psychological variables. Results: In girls, we observed a decrease in BS in the intervention group compared to the control group (β = − 0.101; p = 0.006). In the wave2,, BS decreased more in the intervention group compared to the control group in girls with mothers with a low level of education (β = − 0.338; p = 0.001), with a high level of social support (β = − 0.251; p < 0.001), with greater capacity for conflict resolution (β = − 0.201; p < 0.001) and lower levels of aggressiveness (β = − 0.232, p < 0.001). In boys, the mean levels of HS and BH decreased in wave 2 in both the control and intervention groups. The changes observed after the wave 2 were the same in the control group and in the intervention group. No significant differences were identified between both groups. Conclusions: The implementation of the Lights4Violence was associated with a significant reduction in BS in girls, which highlights the potential of interventions aimed at supporting the personal competencies and social support. It is necessary to reinforce the inclusion of educational contents that promote reflection among boys about the role of gender and the meaning of the attributes of masculinity. Trial registration: Clinicaltrials.gov : NCT03411564. Unique Protocol ID: 776905. Date registered: 26-01-2018.The project “Lights, Camera and Action against Dating Violence” (Ligts4Violence) was funded by the European Commission Directorate-General Justice and Consumers Rights, Equality and Citizen Violence Against Women Program 2016 for the period 2017–2019 to promote healthy dating relationship assets among secondary school students from different European countries, under grant agreement No. 776905. It was also co-supported by the CIBER of Epidemiology and Public Health of Spain for its aid to the Gender-based Violence and Youth Research Program

Calmodulin Supports TRPA1 Channel Association with Opioid Receptors and Glutamate NMDA Receptors in the Nervous Tissue

Supplementary Materials: The following are available online at https://www.mdpi.com/1422-006 7/22/1/229/s1, Figure S1: TRPA1 association with MORs in spinal cord in HINT1-/- and 1R-/- mice. Figure S2: The HINT1 protein or 1R does not support the MOR association with the Nt or Ct regions of TRPA1 channels. Figure S3: CaM mediates the TRPA1 Ct association with MOR in the absence of Ca2+. Figure S4: Pharmacological modulation of TRPA1 associations with MORs and glutamate NMDARs. Figure S5: Formalin-induced inflammatory pain alters TRPA1 associations with MORs and NMDARs. Figure S6: TRPA1 associations with opioid receptors and NMDARs in the CCI model of neuropathic pain.We would like to thank Gabriela de Alba and María José López for their excellent technical assistanceTransient receptor potential ankyrin member 1 (TRPA1) belongs to the family of thermo TRP cation channels that detect harmful temperatures, acids and numerous chemical pollutants. TRPA1 is expressed in nervous tissue, where it participates in the genesis of nociceptive signals in response to noxious stimuli and mediates mechanical hyperalgesia and allodynia associated with different neuropathies. The glutamate N-methyl-d-aspartate receptor (NMDAR), which plays a relevant role in allodynia to mechanical stimuli, is connected via histidine triad nucleotide-binding protein 1 (HINT1) and type 1 sigma receptor (σ1R) to mu-opioid receptors (MORs), which mediate the most potent pain relief. Notably, neuropathic pain causes a reduction in MOR antinociceptive efficacy, which can be reversed by blocking spinal NMDARs and TRPA1 channels. Thus, we studied whether TRPA1 channels form complexes with MORs and NMDARs that may be implicated in the aforementioned nociceptive signals. Our data suggest that TRPA1 channels functionally associate with MORs, delta opioid receptors and NMDARs in the dorsal root ganglia, the spinal cord and brain areas. These associations were altered in response to pharmacological interventions and the induction of inflammatory and also neuropathic pain. The MOR-TRPA1 and NMDAR-TRPA1 associations do not require HINT1 or σ1R but appear to be mediated by calcium-activated calmodulin. Thus, TRPA1 channels may associate with NMDARs to promote ascending acute and chronic pain signals and to control MOR antinociception.MICINN Plan Nacional I+D+i RT 2018-093677B-100University of Granada PPJIB2019.11MECD FPU 15/02356 FPU16/0321

Sex differences in placental protein expression and efficiency in a rat model of fetal programming induced by maternal undernutrition

Fetal undernutrition programs cardiometabolic diseases, with higher susceptibility in males. The mechanisms implicated are not fully understood and may be related to sex differences in placental adaptation. To evaluate this hypothesis, we investigated placental oxidative balance, vascularization, glucocorticoid barrier, and fetal growth in rats exposed to 50% global nutrient re-striction from gestation day 11 (MUN, n = 8) and controls (n = 8). At gestation day 20 (G20), we analyzed maternal, placental, and fetal weights; oxidative damage, antioxidants, corticosterone, and PlGF (placental growth factor, spectrophotometry); and VEGF (vascular endothelial growth factor), 11β-HSD2, p22phox, XO, SOD1, SOD2, SOD3, catalase, and UCP2 expression (Western blot). Compared with controls, MUN dams exhibited lower weight and plasma proteins and higher corticosterone and catalase without oxidative damage. Control male fetuses were larger than female fetuses. MUN males had higher plasma corticosterone and were smaller than control males, but had similar weight than MUN females. MUN male placenta showed higher XO and lower 11β-HSD2, VEGF, SOD2, catalase, UCP2, and feto-placental ratio than controls. MUN females had similar feto-placental ratio and plasma corticosterone than controls. Female placenta expressed lower XO, 11β-HSD2, and SOD3; similar VEGF, SOD1, SOD2, and UCP2; and higher catalase than controls, being 11β-HSD2 and VEGF higher compared to MUN males. Male placenta has worse adaptation to un-dernutrition with lower efficiency, associated with oxidative disbalance and reduced vasculariza-tion and glucocorticoid barrier. Glucocorticoids and low nutrients may both contribute to programming in MUN malesThis research was funded by the Ministerio de Ciencia, Innovación y Universidades (Spain), grant number RTI2018-097504-B-I00, cofinanced with FEDER funds and by the Faculty of Medicine, Khon Kaen University (Thailand), grant number KKU:0514.7.I.12-194