In vitro and in vivo expression of foreign genes by transmissible gastroenteritis coronavirus-derived minigenomes

A helper-dependent expression system based on transmissible gastroenteritis coronavirus (TGEV) has been developed using a minigenome of 3·9 kb (M39). Expression of the reporter gene {beta}-glucuronidase (GUS) (2–8 µg per 106 cells) and the porcine respiratory and reproductive syndrome virus (PRRSV) ORF5 (1–2 µg per 106 cells) has been shown using a TGEV-derived minigenome. GUS expression levels increased about eightfold with the m.o.i. and were maintained for more than eight passages in cell culture. Nevertheless, instability of the GUS and ORF5 subgenomic mRNAs was observed from passages five and four, respectively. About a quarter of the cells in culture expressing the helper virus also produced the reporter gene as determined by studying GUS mRNA production by in situ hybridization or immunodetection to visualize the protein synthesized. Expression of GUS was detected in the lungs, but not in the gut, of swine immunized with the virus vector. Around a quarter of lung cells showing replication of the helper virus were also positive for the reporter gene. Interestingly, strong humoral immune responses to both GUS and PRRSV ORF5 were induced in swine with this virus vector. The large cloning capacity and the tissue specificity of the TGEV-derived minigenomes suggest that these virus vectors are very promising for vaccine development

Constructing TI-Friendly Substitution Boxes Using Shift-Invariant Permutations

The threat posed by side channels requires ciphers that can be efficiently protected in both software and hardware against such attacks. In this paper, we proposed a novel Sbox construction based on iterations of shift-invariant quadratic permutations and linear diffusions. Owing to the selected quadratic permutations, all of our Sboxes enable uniform 3-share threshold implementations, which provide first order SCA protections without any fresh randomness. More importantly, because of the shift-invariant property, there are ample implementation trade-offs available, in software as well as hardware. We provide implementation results (software and hardware) for a four-bit and an eight-bit Sbox, which confirm that our constructions are competitive and can be easily adapted to various platforms as claimed. We have successfully verified their resistance to first order attacks based on real acquisitions. Because there are very few studies focusing on software-based threshold implementations, our software implementations might be of independent interest in this regard

Shock absorbing reception surfaces for collecting fruit during the mechanical harvesting of citrus

Damage to fresh citrus caused by impact of fruits onto collecting surfaces has restricted the adoption of mechanical harvesting. Two different experiments were carried out: investigating free-falling citrus and investigating the shock absorbing capacity of various surfaces. In free-falling experiment damage to mandarin, orange and lemon was studied. Three collecting surfaces were studied: a concrete floor, an elevated canvas provided with a frame and wheels, and a concrete floor covered with a shock absorbing canvases. Three dropping heights were used. In the shock absorbing experiment, an electronic sphere and a triaxial accelerometer were used to measure the shock capacity of seven receiving surfaces: earth, earth covered with a shock absorbing canvas, earth covered with weeds, earth covered with a mulch and an elevated canvas provided with a frame and wheels. The elevated canvases had a higher shock absorbing capacity compared to the other surfaces (260 m s−2 maximum acceleration compared with 1753 m s−2 to 2772 m s−2). Weeds, mulch and the shock absorbing canvases showed significantly higher shock absorbing capacity than the bare earth. Also, the shock absorbing canvas covering the concrete floor reduce impact and fruit damage (1866 m s−2 maximum acceleration compared to 2477 m s−2). Citrus damage susceptibility during harvest depended on variety. Weeds, mulch and shock absorbing canvases were shown to reduce impact when they cover earth during the mechanical harvesting of citrus. Elevated canvases could be used as collection systems for the mechanical harvesting of fresh citrus.This study was funded by the Ministerio de Ciencia e Innovacion (research project RTA2009-00118-C02-02) and FEDER. The authors are most grateful to the following collaborators: Maria del Mar Lopez Quevedo, Montano Perez, Juan Jose Pena and Angel Perez.Ortiz Sánchez, MC.; Blasco, J.; Balasch Parisi, S.; Torregrosa Mira, A. (2011). Shock absorbing reception surfaces for collecting fruit during the mechanical harvesting of citrus. Biosystems Engineering. 110(1):2-9. https://doi.org/10.1016/j.biosystemseng.2011.05.006S29110

Actuación en zonas antiguas de pueblos y ciudades

Actuación en zonas antiguas de pueblos y ciudade

Milk yield estimation during suckling using the double oxytocin injection-milking and the double weighing-suckling methods in dairy goats

The aim was to verify the validity of the double oxytocin-milking (DOT) method as milk yield estimator during the suckling period of lactating dairy goats. To this end, it was necessary to determine whether the weighing suckling weighing (WSW) and DOT methods of milk yield estimation satisfied the criteria to be considered valuable, the accuracy between both methods and the suitability of DOT to evaluate actual milk. At parturition, sixty lactating Murciano Granadina breed goats were separated into 2 groups, in mixed (MS; n = 24) and artificial rearing (ARS; n = 36) management systems. Until the sixth week of lactation (weaning), MS goats suckled one kid while kids from ARS goats were artificially reared; moreover, goats in both systems were submitted to once-a-day milking. Once per week, actual milk yield for ARS goats and potential milk yield were recorded using DOT method for all goats, except for 12 goats in ARS which remained as a control. Twelve goats from each management system were used to evaluate diurnal variation in milk production (DVM) by DOT method for 6 consecutive days in week 4 of lactation. No difference in DVM was found by DOT method in 4-h milk production of goats in MS (P = 0.099) or ARS (P = 0.220), which allowed sixfold multiplication of milked milk volume to obtain potential milk per day. ARS goats subjected to a weekly DOT and control group goats showed a similar (P = 0.379) lactation curve for the first 6 weeks of lactation. The DOT method slightly overestimated (3.4%, P = 0.005) the milk yield evaluated by WSW method for goats under an MS, but fitted the actual milk obtained by common milk records for the group of goats in an ARS submitted to the DVM test (P = 0.357) and the group in ARS alone (P = 0.163). The DOT method applied for 8 consecutive days led to a drop of 6 12% in milk yield during the following week for both production systems. In conclusion, DOT was an accurate method to estimate milk yield during the first weeks of lactation both in MS and ARS under the conditions of this experiment. © 2013 Elsevier B.V. All rights reserved.Fernández Martínez, N.; Balasch Parisi, S.; Pérez Baena, I.; Rodríguez Garcia, M.; Peris Ribera, CJ. (2013). Milk yield estimation during suckling using the double oxytocin injection-milking and the double weighing-suckling methods in dairy goats. Small Ruminant Research. 112(1-3):181-185. doi:10.1016/j.smallrumres.2012.12.023S1811851121-

Machine milking parameters for Murciano-Granadina breed goats

[EN] In dairy ruminants, the combination of milking parameters must ensure good milking performance without harming udder conditions. Commonly, milking conditions for goats are established without having checked the admissible limits for optimal and fast milking. The aim of this study was to establish a limit combination of machine milking parameters that improves machine milking performance without altering milkability or udder status. To this end, we studied the effect of 2 combinations (42 kPa, 120 cpm, 60% vs. 44 kPa, 120 cpm, 60% in terms of kilopascals of vacuum level, cycles per minute of pulsator rate, and percentage of pulsator ratio, respectively) on milk production and composition, milk fractioning during milking, SCC, teat tissue thickness variation after milking, and the milk emission kinetics parameters throughout 1 lactation period (6 rno). The 42 and 44 kPa measured at the vacuum gauge level became average values of 37.5 and 39.3 kPa, respectively, measured at the teat sphincter level during milking. Milk flow significantly increased and total milking time decreased 25 s with the elevation of the vacuum level from 42 to 44 kPa without any adverse effect on milk fractioning at milking. However, the use of 44 kPa also showed an increase in tissue thickness above 5%, and we observed a tendency of average conductivity of milk to increase, although without any adverse effect on SCC. It seems that 44 kPa, 120 cpm, 60% is a possible limit combination of parameters to improve milking performance without altering milkability or udder conditions. We concluded that this combination can be used for milking Murciano-Granadina breed goats in conditions similar to those of this study (mid-level milking system and 1 milking/d), although further studies are necessary to verify its application in the case of 2 milkings/d.Fernández Martínez, N.; Martí Vicent, JV.; Rodríguez Garcia, M.; Peris Ribera, CJ.; Balasch Parisi, S. (2020). Machine milking parameters for Murciano-Granadina breed goats. Journal of Dairy Science. 103(1):507-513. https://doi.org/10.3168/jds.2019-16446S5075131031Alejandro, M., Roca, A., Romero, G., & Díaz, J. R. (2014). Short communication: Effects of milk removal on teat tissue and recovery in Murciano-Granadina goats. Journal of Dairy Science, 97(8), 5012-5016. doi:10.3168/jds.2014-7934Ali, A. K. A., & Shook, G. E. (1980). An Optimum Transformation for Somatic Cell Concentration in Milk. Journal of Dairy Science, 63(3), 487-490. doi:10.3168/jds.s0022-0302(80)82959-6Blasco, E., Gomez, E. A., Vicente, C., Vidal, G., & Peris, C. (2016). Factors affecting milking speed in Murciano-Granadina breed goats. Journal of Dairy Science, 99(12), 10102-10108. doi:10.3168/jds.2016-10869BOE (Boletín Oficial del Estado). 2013. Real Decreto 53/2013, de 1 de febrero, por el que se establecen las normas básicas aplicables para la protección de los animales utilizados en experimentación y otros fines científicos, incluyendo la docencia.Delgado-Pertíñez, M., Guzmán-Guerrero, J. L., Mena, Y., Castel, J. M., González-Redondo, P., & Caravaca, F. P. (2009). Influence of kid rearing systems on milk yield, kid growth and cost of Florida dairy goats. Small Ruminant Research, 81(2-3), 105-111. doi:10.1016/j.smallrumres.2008.12.007Díaz, J. R., Alejandro, M., Peris, C., & Fernández, N. (2013). Use of ultrasound scanning to estimate teat wall thickness in Murciano-Granadina goats. Livestock Science, 155(1), 114-122. doi:10.1016/j.livsci.2013.04.007Díaz, J. R., Romero, G., Muelas, R., Alejandro, M., & Peris, C. (2012). Effect of intramammary infection on milk electrical conductivity in Murciano-Granadina goats. Journal of Dairy Science, 95(2), 718-726. doi:10.3168/jds.2011-4698Diaz, J. R., Romero, G., Muelas, R., Sendra, E., Pantoja, J. C. F., & Paredes, C. (2011). Analysis of the influence of variation factors on electrical conductivity of milk in Murciano-Granadina goats. Journal of Dairy Science, 94(8), 3885-3894. doi:10.3168/jds.2011-4187Fernández, N., Martínez, A., Martí, J. V., Rodríguez, M., & Peris, C. (2015). Milkability and milking efficiency improvement in Murciano-Granadina breed goats. Small Ruminant Research, 126, 68-72. doi:10.1016/j.smallrumres.2015.03.001Gil, Z. (1988). Milk temperature fluctuations during milking in cows with subclinical mastitis. Livestock Production Science, 20(3), 223-231. doi:10.1016/0301-6226(88)90074-7Hamann, J., Mein, G. A., & Nipp, B. (1996). Recommended method for measuring changes in thickness of the bovine teat with spring-loaded calipers. Journal of Dairy Research, 63(2), 309-313. doi:10.1017/s0022029900031800Hamzaoui, S., Salama, A. A. K., Albanell, E., Such, X., & Caja, G. (2013). Physiological responses and lactational performances of late-lactation dairy goats under heat stress conditions. Journal of Dairy Science, 96(10), 6355-6365. doi:10.3168/jds.2013-6665Isaksson, A., & Lind, O. (1992). Teat Reactions in Cows Associated With Machine Milking*. Journal of Veterinary Medicine Series A, 39(1-10), 282-288. doi:10.1111/j.1439-0442.1992.tb00184.xLE DU, J., BENMEDERBEL, B., DANO, Y., COMBAUD, J. F., & TESSONNIÈRE, R. (1984). Aptitude des chèvres de race Saanen à la traite mécanique. Relations avec les caractéristiques physiques du trayon. Annales de Zootechnie, 33(3), 375-384. doi:10.1051/animres:19840307Peris, S., Caja, G., Such, X., Casals, R., Ferret, A., & Torre, C. (1997). Influence of Kid Rearing Systems on Milk Composition and Yield of Murciano-Granadina Dairy Goats. Journal of Dairy Science, 80(12), 3249-3255. doi:10.3168/jds.s0022-0302(97)76299-4Requena, R., Balasch, S., Peris, C., Rodríguez, M., & Fernández, N. (2010). Dose response of lactating dairy ewes during suckling and milking to bovine somatotropin1. Journal of Animal Science, 88(9), 3136-3144. doi:10.2527/jas.2009-2690Zecconi, A., Hamann, J., Bronzo, V., & Ruffo, G. (1992). Machine-induced teat tissue reactions and infection risk in a dairy herd free from contagious mastitis pathogens. Journal of Dairy Research, 59(3), 265-271. doi:10.1017/s002202990003054

The scope of language contact as a constraint factor in language change: The periphrasis haber de plus infinitive in a corpus of language immediacy in modern Spanish

In this work an empirical study grounded in the principles and methods of the comparative variationist framework is conducted to measure the scope of language contact as a factor constraining some potentially diverging uses of a Spanish verbal periphrasis that has undergone a sharp decline over the last century (haber de plus infinitive). The analysis is based on three independent samples of text that correspond to three dialectal areas of peninsular Spanish (monolingual zones, Catalan-speaking linguistic territories and the north-western linguistic area). These samples, extracted from a corpus made up of texts of communicative immediacy from the 19th and the first half of the 20th centuries, confirm the existence of a certain linguistic convergence in the expressive habits of the speakers in the bilingual communities. In each region, however, the outcomes are different, due to parallel differences in the structural position of the periphrasis in each language. However, a thorough analysis of the variable context that surrounds the periphrasis shows that the observed differences do not affect the essence of the underlying grammar of this variant, whose decline (which favours tener que plus infinitive and becomes faster as the 20th century advances) is constrained by identical linguistic and extralinguistic conditioning factors in all the dialectal areas

RNA-Seq Reveals an Integrated Immune Response in Nucleated Erythrocytes

Background: Throughout the primary literature and within textbooks, the erythrocyte has been tacitly accepted to have maintained a unique physiological role; namely gas transport and exchange. In non-mammalian vertebrates, nucleated erythrocytes are present in circulation throughout the life cycle and a fragmented series of observations in mammals support a potential role in non-respiratory biological processes. We hypothesised that nucleated erythrocytes could actively participate via ligand-induced transcriptional re-programming in the immune response. Methodology/Principal Findings: Nucleated erythrocytes from both fish and birds express and regulate specific pattern recognition receptor (PRR) mRNAs and, thus, are capable of specific pathogen associated molecular pattern (PAMP) detection that is central to the innate immune response. In vitro challenge with diverse PAMPs led to de novo specific mRNA synthesis of both receptors and response factors including interferon-alpha (IFNα) that exhibit a stimulus-specific polysomal shift supporting active translation. RNA-Seq analysis of the PAMP (Poly (I:C), polyinosinic:polycytidylic acid)-erythrocyte response uncovered diverse cohorts of differentially expressed mRNA transcripts related to multiple physiological systems including the endocrine, reproductive and immune. Moreover, erythrocyte-derived conditioned mediums induced a type-1 interferon response in macrophages thus supporting an integrative role for the erythrocytes in the immune response. Conclusions/Significance: We demonstrate that nucleated erythrocytes in non-mammalian vertebrates spanning significant phylogenetic distance participate in the immune response. RNA-Seq studies highlight a mRNA repertoire that suggests a previously unrecognized integrative role for the erythrocytes in other physiological systems

Swedish high school students' knowledge and attitudes regarding fertility and family building

<p>Abstract</p> <p>Background</p> <p>Infertility is a serious problem for those who suffer. Some of the risks for infertility are preventable and the individual should therefore have knowledge of them. The purposes of this study were to investigate high-school students' knowledge about fertility, plans for family building and to compare views and knowledge between female and male students.</p> <p>Methods</p> <p>A questionnaire containing 34 items was answered by 274 students. Answers from male and female students were compared using student's <it>t</it>-test for normally distributed variables and Mann-Whitney <it>U</it>-test for non-normal distributions. The chi-square test was used to compare proportions of male and female students who answered questions on nominal and ordinal scales. Differences were considered as statistically significant at a <it>p</it>-value of 0.05.</p> <p>Results</p> <p>Analyses showed that 234 (85%) intended to have children. Female students felt parenthood to be significantly more important than male students: <it>p </it>= <it><</it>0.01. The mean age at which the respondents thought they would like to start to build their family was 26 (± 2.9) years. Men believed that women's fertility declined significantly later than women did: <it>p </it>= <it><</it>0.01. Women answered that 30.7% couples were involuntarily infertile and men answered 22.5%: <it>p </it>= <it><</it>0.01. Females thought it significantly more likely that they would consider IVF or adoption than men, <it>p = </it>0.01. Men felt they were more likely to abstain from having children than women: <it>p = <</it>0.01. Women believed that body weight influenced fertility significantly more often than men: <it>p = <</it>0.01 and men believed significantly more often that smoking influenced fertility: <it>p </it>= 0.03. Both female and male students answered that they would like to have more knowledge about the area of fertility.</p> <p>Conclusions</p> <p>Young people plan to start their families when the woman's fertility is already in decline. Improving young people's knowledge about these issues would give them more opportunity to take responsibility for their sexual health and to take an active role in shaping political change to improve conditions for earlier parenthood.</p