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EL SARS-CoV-2 es el agente etiológico responsable de la enfermedad COVID-19 que inició una pandemia desde finales de 2019. Existen distintos modelos animales para esta enfermedad siendo el modelo de ratón transgénico K18-hACE2, originalmente desarrollado para el estudio del SARS-CoV-1, de gran relevancia en el contexto actual. Considerando que los ratones se comercializan como hemicigotas y dada la ausencia de ratones homocigotas para el transgén K18-hACE2, el objetivo de este trabajo fue generar una colonia de ratones homocigotas K18-hACE2-Tg/Tg bajo condiciones ambientales controladas. Para la F1 se utilizaron 10 ratones hemicigotas (7 hembras y 3 machos), de 6-8 semanas de edad, provenientes de The Jackson Laboratory (USA). De la F1 se identificaron mediante genotipificación por PCR 78 animales: 62 hemicigotas (Tg/0) y 16 salvajes (0/0). Aquellos animales identificados como transgénicos se cruzaron con ratones C57BL/6J para seleccionar los parentales homocigotas que generaron 100% de hemicigotas en la F2. De 224 crías analizadas, se obtuvieron 7 parentales homocigotas K18-hACE2-Tg/Tg que fueron utilizados para la colonia fundación de ratones homocigotas K18-hACE2-Tg/Tg. El modelo de ratón homocigota K18-hACE2-Tg/Tg desarrollado en este trabajo podrá ser empleado para el estudio de la patogénesis de la enfermedad y la evaluación de posibles terapéuticos contra el SARS-CoV-2

En este estudio se evaluó en una prueba de concepto, la seguridad y funcionalidad del equipo prototipo Ciclador Automático para Resucitador (CAR) en un cerdo de peso similar al de un humano adulto. Para ello, se estudiaron los volúmenes corrientes de 5.8, 7 y 9.7 ml/kg para las presiones positivas al final de la expiración (PEEP) de 5, 10 y 15 cmH2O. Los parámetros indicativos del estado homeostático del animal se obtuvieron a través de un equipo multiparamétrico y un sistema de análisis bioquímicos de sangre EPOC®. En los periodos en los que se evaluaron los volúmenes corrientes de 7 y 9.7 ml/kg los parámetros se mantuvieron dentro de rangos fisiológicos normales. Sin embargo, el volumen corriente de 5.8 ml/kg no fue suficiente para mantener una ventilación adecuada lo que se manifestó a través de hipercapnia y caída del pH. Como conclusión general se puede inferir que con adaptaciones en los patrones de ventilación (7 y 9.7 ml/kg), la utilización del prototipo CAR permitió mantener al animal con vida durante 24 h conservando su homeostasis.

Background: Hemolytic uremic syndrome associated with Shiga-toxin produced by Escherichia coli is a serious worldwide foodborne disease. Nowadays, no treatment is available, only supportive care can be provided, and 50 % of the patients require a period of dialysis. Recently, a therapy based on Neutralizing Equine Anti Shiga Toxin (NEAST) antibodies has been developed. NEAST is composed of F(ab’)2 fragments from equine immunoglobulins. Objective: The purpose of this study was to develop an ELISA to measure serum concentrations of NEAST in mice and rabbits, and to validate it according to international recommendations. The validated method was further used to analyze the NEAST PK during preclinical studies. Methods: A sandwich ELISA was developed, the performance of the calibration curve was assessed, and it was validated based on the parameters as accuracy, precision, specificity, selectivity, stability of the analyte, and dilutional linearity. Results: This immunoassay was specific, sensitive, accurate and precise in a dynamic range from 7.81 to 500 ng/mL and from 15.63 to 500 ng/mL for mice and rabbits, respectively. This method was successfully applied to PK studies of NEAST after intravenous administration. conclusion: The results obtained are expected for a robust ELISA used for macromolecule analysis. Since NEAST is an equine F(ab′)2, this immunoassay would serve for the evaluation of the PK profile of any biological product based on molecules with similar characteristics. This immunoassay may be useful for current and future preclinical trials conducted for registration purposes.

Heat stress has been widely studied in relation to its effects on the production and reproduction of dairy cattle. However, the long-term effects of heat stress during intrauterine development on adult cows have been scarcely considered. Thus, the aim of this study was to evaluate possible changes in the reproductive performance of Holstein cows gestated under different values of the Temperature-Humidity Index (THI) during their intrauterine development. Data collected from a database of reproductive and productive records of 10,790 Holstein cows from the central region of Argentina and the THI data from the agrometeorological station of the Instituto Nacional de Tecnología Agropecuaria (INTA) EEA Rafaela, Santa Fe, Argentina, were used. The gestation of the cows evaluated was divided into trimesters, in which the highest THI cycles (sum of three or more consecutive days of exposure to a THI ≥72 during each trimester) and number of days with a THI ≥ 72 were calculated. The calving-to-conception and calving-to-first-service intervals of the cows evaluated were considered as reproductive variables associated with their first lactation. Generalized Linear Mixed Models were used, considering the cow's dairy farm as a random factor within the model. The exposure of the animals to environments with a THI ≥72 during the first trimester of gestation had a negative impact on the reproductive efficiency parameters analyzed. The results obtained indicate that the exposure of pregnant females to high THI values has a long-term impact on their daughters, which may contribute to a decrease in their reproductive performance, possibly through inherited epigenetic characteristics that remain in later generations through fetal programming.

Reproductive diseases in dairy cows as cystic ovarian disease (COD) represent a major problem that impacts on dairy production. COD is characterized by anovulation, persistence of the dominant follicle, and interruption of normal estrous cycles. Anovulation is attributable to a failure in the LH surge, due to endocrine imbalances and alterations in local factors, such as the insulin signaling pathway. Thus, the aim of this study was to examine the expression of critical nodes of the insulin pathway, including insulin receptor (IR), IR substrate (IRS), phosphatidylinositol-3-kinase (PI3K), and protein kinase B (also known as Akt or pan-Akt), in ovarian follicular structures of cows during the development of follicular persistence induced by long-term progesterone administration. Immunoexpression of IR, p-IR, IRS1, p-IRS1, PI3K, pan-Akt and p-pan-Akt was evaluated in situ by immunohistochemistry and the concentration of insulin in serum and follicular fluid was determined by radioimmunoassay. p-IR, p-IRS1, PI3K and p-pan-Akt expression was decreased in follicles at different times of persistence in relation to the control dominant follicles, in both granulosa and theca cells, whereas IR and IRS1 immunoexpression was decreased in persistent follicles at 5 and 15 days of persistence in granulosa cells. Serum and follicular fluid insulin concentration was higher in cows with persistent follicles than in control cows. These results show that decreased expression and/or activation of the receptors and other intermediates of the insulin signaling pathway in persistent follicles indicates that reduced response/resistance to insulin rather than the concentration of insulin per se may be one of the important molecular mechanisms in the development of persistent follicles in dairy cows.

Hemolytic uremic syndrome associated to Shiga-toxin produced by Escherichia coli is a serious worldwide foodborne disease. Nowadays, no treatment is available, only supportive care can be provided, and 50 % of the patients require a period of dialysis. Recently, a therapy based on Neutralizing Equine Anti Shiga Toxin (NEAST) antibodies has been developed. NEAST is composed of F(ab’)2 fragments from equine immunoglobulins. In order to conduct preclinical studies of NEAST, a quantitative bioanalytical method based on a sensitive enzyme-linked immunosorbent assay (ELISA) was developed to quantify specific F(ab’)2 in mouse and rabbit serum, and validated in accordance with FDA and EMA guidelines. This immunoassay has been demonstrated to be robust, reproducible and reliable to determine NEAST levels, and is in use in several current and upcoming preclinical trials conducted for registration purposes. Furthermore, since NEAST is an equine F(ab′)2, the developed ELISA would prove useful for the evaluation of the pharmacokinetic profile of any biological product based on molecules with similar characteristics.

Context Dairy cattle experience stressful environmental situations that affect production. Heat stress during gestation can influence the intrauterine development of offspring, resulting in long-term damage that can affect the reproductive life of the adult offspring. Aim The aim of the present study was to evaluate changes in the expression and regulation of steroid hormone receptors in the ovary of Holstein cows gestated under different temperature-humidity index (THI) during their in utero development. Methods Animals were classified by their exposure to temperature-humidity index (THI) ≥72 during their development in utero according to date of birth or date of effective service of their mother. This study was not carried out under controlled conditions, but the conditions to which the cows were naturally exposed during their development were considered retrospectively, controlling the variables in the statistical analyses (age as a covariate, dairy farm as a random factor). Gestation was divided into two periods (P1 = days 0–150; and P2 = day 151 to calving) and three trimesters (T1 = days 0–90; T2 = days 91–180; and T3 = day 181 to calving), and the exposure to THI ≥72 was calculated in each one. The following characteristics were evaluated: gene expression of estrogen receptor (ESR) 1, ESR2 and progesterone receptor (PGR), CpG methylation in the 5′UTR of ESR1 and ESR2, and protein expression of ESR1, ESR2, PGR and coregulatory proteins in the dominant follicles of daughter cows in adulthood. Key results We found associations between heat stress variables during gestation and the methylation status of CpG sites in the 5′UTR of ESR1 and ESR2 in dominant follicles. Results also showed association between exposure to high THI values during intrauterine development and expression of ESR1, ESR2 and PGR and coregulatory proteins in dominant follicles of adult cows. Conclusions These results provide novel information about the impact of prenatal heat stress on molecular aspects at the ovary level in the offspring, during their adult life, which probably impacts the reproductive aspects of the herd.

Native potatoes are the most diverse among cultivated potato species and thus constitute a valuable source for identifying genes for potato improvement. Nevertheless, high-density mapping, needed to reveal allelic diversity, has not been performed for native Argentinian potatoes. We present a study of the genetic variability and population structure of 96 Andigena potatoes from Northwestern Argentina performed using a subset of 5035 SNPs with no missing data and full reproducibility. These high-density markers are distributed across the genome and present a good coverage of genomic regions. A Bayesian approach revealed the presence of: (I) a major group comprised of most of the Andean accessions; (II) a smaller group containing the out-group cv. Spunta and the sequenced genotype DM; and (III) a third group containing colored flesh potatoes. This grouping was also consistent when maximum likelihood trees were constructed and further confirmed by a principal coordinate analysis. A group of 19 accessions stored as Andean varieties clustered consistently with group Tuberosum accessions. This was in agreement with previous studies and we hypothesize that they may be reintroductions of European-bred long day-adapted potatoes. The present study constitutes a valuable source for allele mining of genes of interest and thus provides a tool for association mapping studies.

Aspartic proteases are proteolytic enzymes widely distributed in living organisms and viruses. Although they have been extensively studied in many plant species, they are poorly described in potatoes. The present study aimed to identify and characterize

In dairy cows, reproductive diseases such as cystic ovarian disease (COD) represent a major problem that impacts on dairy production. It has been postulated that the insulin-like growth factor (IGF) system may contribute to follicular persistence and development of COD. Thus, the aim of the present study was to analyze relevant members of the IGF system in a critical period immediately after the expected time of ovulation, to obtain information about their role in follicular persistence in dairy cows. Proteins of the IGF system were evaluated at 0 (expected day of ovulation), 5, 10 and 15 days of follicular persistence to determine whether the changes previously detected in cows with COD occur early in COD pathogenesis. The serum concentration of IGF1 was higher in cows with 10 and 15 days of follicular persistence than in control cows. IGF1 expression in granulosa cells was similar in the follicles analyzed. In contrast, in theca cells, persistent follicles of days 5 and 10 showed the lowest IGF1 expression. IGF binding protein (IGFBP) 2 and 3 expression was lower in persistent follicles than in dominant follicles of the control group. Although IGF receptor (IGFR) 1 expression was similar in the groups analyzed, p-IGFR1 expression was significantly higher in dominant follicles of the control group than in persistent follicles. These data suggest alterations in the IGF system at the early stages of follicular persistence. The evidences obtained allow supporting that the IGF system could plays a key role in dairy cattle reproduction.

Throughout the estrous cycle the mammalian endometrium undergoes morphological and functional changes that are essential for the establishment of pregnancy and proper ovarian and uterine functions. Among these changes, the most important are alterations in both inter- and intracellular signalling molecules, many of which modulate immune processes. In the endometrial tissue there are local innate (nonspecific) and adaptive (specific/acquired) response mechanisms which vary because of the endocrine status during the estrous cycle, pregnancy and postpartum period. Endometrial cells have responses that support the immune system by producing pro-inflammatory factors such as cytokines, sensors, effector molecules and chemokines. This response is important during gestation, pregnancy, and fetal growth, as well as in preventing infection, and immuno-rejection of the semi-allogeneic embryo. In dairy cows, both before and immediately after calving, there are marked changes in the values for hormonal and metabolic variables and the immune status is impaired. Thus, in several studies there has been assessment of the physiological and/or abnormal maternal immune changes and possible effects on dairy cow reproductive performance. The objective with this review is to summarize the novel information about the immune mechanisms involved during the postpartum period, subsequent peri-implantation period and pregnancy in dairy cows, and the possible effects on reproductive performance. This information provides for an enhanced understanding of the local and systemic immune responses associated with the metabolic and hormonal status of dairy cows, and alterations in the immune system of high producing cows and the possible effects on subsequent fertility. Fil: Velázquez, Melisa María del Luján. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Peralta, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Angeli, Emmanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Stassi, Antonela Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Gareis, Natalia Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Durante, Leandro Ivan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Cainelli, Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

The objective of this study was to examine the expression of transforming growth factor beta receptor (TGFBR)1, TGFBR2, TGFBR3, activin receptor (ACVR)1B and ACVR2B in ovaries of cows with cystic ovarian disease (COD). The expression of the selected receptors was determined by immunohistochemistry in sections of ovaries from cows with ACTH-induced and spontaneous COD. Expression of TGFBR1 and TGFBR3 was higher in granulosa cells of cysts from cows with spontaneous COD than in tertiary follicles from the control group. Additionally, TGFBR3 expression was higher in granulosa cells of cysts from cows with ACTH-induced COD than in those from the control group and lower in theca cells of spontaneous and ACTH-induced cysts than in tertiary control follicles. There were no changes in the expression of TGFBR2. ACVR1B expression was higher in granulosa cells of tertiary follicles of cows with spontaneous COD than in the control group, whereas ACVR2B expression was higher in cysts of the spontaneous COD group than in tertiary follicles from the control group. The alterations here detected, together with the altered expression of the ligands previously reported, indicate alterations in the response of the ligands in the target cells, modifying their actions at cellular level. Fil: Matiller, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Hein, Gustavo Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Stassi, Antonela Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Angeli, Emmanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Belotti, Eduardo Matías. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

The aim of this study was to investigate the role of tocopherols in the susceptibility of Star Ruby grapefruit to postharvest chilling injury (CI). Fruit exposed to normal sunlight (NC, non-covered) and deprived of light (C, covered) in the last stages of development were used. Tocopherol contents increased in the flavedo of both NC and C fruit during development, concomitantly with the up-regulation of TAT1 and most genes of the tocopherol-core pathway. Fruit shading reduced total contents by repressing γ-tocopherol accumulation, associated to a down-regulation of GGDR and VTE1 and, to a lesser extent, of VTE2, VTE3a and VTE4. During cold storage, total and α-tocopherol contents increased in NC and C fruit, and no direct relationship between tocopherol accumulation and CI tolerance was found. Cold stress up-regulated most genes involved in the synthesis of tocopherol precursors and down-regulated those of the tocopherol-core pathway, but changes seemed to be cold-mediated and not related to CI development., This work was supported by research grants RTI2018-095131-B-I00 of the Ministry of Science and Innovation (Spanish Government). F. Rey is the recipient of a PhD scholarship (POS_EXT_2016_1_133720) from ANII (Uruguay).

Plants are sessile organisms that, to survive they develop response mechanisms under water deficit conditions. Plant proteases play an essential role in a diversity of biological processes, among them tolerance to drought stress. Proteolysis is a critical regulator of stomatal development. Plant proteases are involved in the crosstalk among phytohormones and adjustment of stomatal aperture. Plant proteases are also related to the increment in reactive oxygen species (ROS) production detected in the plant biochemical response to drought. Plant proteases mitigate this process by degrading damaged, denatured, and aggregated proteins, remobilizing amino acids, and generating molecules involved in signal transductions. Although many roles for proteases have been proposed, molecular bases that regulate these mechanisms remain unknown. In this review, we summarize the current knowledge on the participation of proteases in the signaling pathways of plants in response to water deficit and their relationship with plant stress tolerance.

Stressors activate the hypothalamic-pituitary-adrenal (HPA) axis, reducing fertility by interfering with the mechanisms that regulate the timing of events within the follicular phase of the estrous cycle. In the HPA axis, melanocortin 2 receptor (MC2R) mediates responses to adrenocorticotropic hormone (ACTH) in concert with melanocortin receptor accessory protein 2 (MRAP2). The aims of the present study were: (1) to evaluate the effects of ACTH administered in cows in the preovulatory period on the expression of the MC2R/MRAP2 complex in the dominant follicle; and (2) to analyze the involvement of Extracellular signal Regulated Kinase 1 (ERK1) signaling in the activation of MC2R and the expression of key enzymes involved in the biosynthesis of glucocorticoids (GCs) in the dominant follicle. To this end, 100 IU ACTH was administered to Holstein cows from a local dairy farm during pro-estrus every 12 h for four days until ovariectomy, which was performed before ovulation. Protein immunostaining of MC2R was higher in the dominant follicles of ACTH-treated cows (p < 0.05). Also, Western blot analysis showed higher activation of the ERK1 signaling pathway in ACTH-treated cows (p < 0.05). Finally, immunohistochemistry performed in the dominant follicles of ACTH-treated cows detected higher expression of CYP17A1 and CYP21A2 (p < 0.05). These results suggest that the bovine ovary is able to respond locally to ACTH as a consequence of stress altering the expression of relevant steroidogenic enzymes. The results also confirm that the complete GC biosynthesis pathway is present in bovine dominant follicle and therefore GCs could be produced locally.

Citrus fruit are sensitive to chilling injury (CI) during cold storage, a peel disorder that causes economic losses. C-repeat binding factors (CBFs) are related to cold acclimation and tolerance in different plants. To explore the role of Citrus CBFs in fruit response to cold, an in silico study was performed, revealing three genes (CBF1, CBF2, and CBF3) whose expression in CI sensitive and tolerant cultivars was followed. Major changes occurred at the early stages of cold exposure (1–5 d). Interestingly, CBF1 was the most stimulated gene in the peel of CI-tolerant cultivars (Lisbon lemon, Star Ruby grapefruit, and Navelina orange), remaining unaltered in sensitive cultivars (Meyer lemon, Marsh grapefruit, and Salustiana orange). Results suggest a positive association of CBF1 expression with cold tolerance in Citrus cultivars (except for mandarins), whereas the expression of CBF2 or CBF3 genes did not reveal a clear relationship with the susceptibility to CI. Light avoidance during fruit growth reduced postharvest CI in most sensitive cultivars, associated with a rapid and transient enhance in the expression of the three CBFs. Results suggest that CBFs-dependent pathways mediate at least part of the cold tolerance responses in sensitive Citrus, indicating that CBF1 participates in the natural tolerance to CI., This work was supported by a research grant from the National Agency of Research and Innovation (ANII-Uruguay) by the code FCE_3_2016_1_126714 and by a research grant RTI2018–095131-B-I00 of the Ministry of Science and Innovation (Spanish Government). F. Rey is the recipient of a predoctoral scholarship (POS_EXT_2016_1_133720) from ANII (Uruguay).

Tocopherols are plant-derived isoprenoids with potent antioxidant activity, which have been implicated in the tolerance of plants to different stresses. However, tocopherol accumulation and biosynthesis in fruit, and their potential implication in postharvest chilling injury (CI), has been scarcely studied. Therefore, in this work, we have investigated tocopherol accumulation and biosynthesis in the peel of mandarin fruit of three cultivars with contrasting susceptibility to CI during storage at 2 °C (‘Fortune’>‘Nova’> ‘Nadorcott’). α- and γ-tocopherol were the isoforms detected in the flavedo of the fruit, and a direct relationship between tocopherols content and CI-tolerance was found, since CI-tolerant fruit accumulated the highest tocopherol content whereas CI-sensitive fruit the lowest. Moreover, the transcriptional profiling of 14 genes related to the specific steps of tocopherol biosynthesis, and to their precursor’s synthesis, were analyzed. Upstream genes DXS1 and DXS2 (1-deoxy-d-xylulose-5-phosphate synthase) and GGDR (geranylgeranyl diphosphate reductase), involved in the supply of phytyl pyrophosphate, and the VTE3 (2-methyl-6-phytyl-1,4-benzoquinol methyltransferase) isoforms appear to be key for the differences in total tocopherol content among the cultivars at harvest. During cold storage, most genes involved in the precursors supply were up-regulated, whereas genes of the tocopherol-core pathway were in general repressed. Changes in VTE4 during cold storage may account for the differences in γ-tocopherol among cultivars. Collectively, results suggest that the concentration of tocopherols at harvest may play a function in the natural tolerance of mandarin fruit to CI, and that changes in the expression of genes during storage appear to be cold-regulated responses, rather than involved in CI tolerance., This work was funded by the research grant RTI2018-095131-B-I00 of the Ministry of Science and Innovation (Spanish Government). F.R. is the recipient of a PhD scholarship (POS_EXT_2016_1_133720) from ANII (Uruguay)

Citrus fruit is one of the most important contributors to the ascorbic acid (AsA) intake in humans. Here, we report a comparative analysis of AsA content and transcriptional changes of genes related to its metabolism during development of petals, leaves and fruits of Valencia Late oranges (Citrus sinensis). Petals of close flowers and at anthesis contained the highest concentration of AsA. In fruits, AsA content in the flavedo reached a maximum at color break, whereas the pulp accumulated lower levels and experienced minor fluctuations during development. AsA levels in leaves were similar to those in the flavedo at breaker stage. The transcriptional profiling of AsA biosynthetic, degradation, and recycling genes revealed a complex and specific interplay of the different pathways for each tissue. The D-galacturonic acid pathway appeared to be relevant in petals, whereas in leaves the L-galactose pathway (GGP and GME) also contributed to AsA accumulation. In the flavedo, AsA content was positively correlated with the expression of GGP of the L-galactose pathway and negatively with DHAR1 gene of the recycling pathway. In the pulp, AsA appeared to be mainly controlled by the coordination among the D-galacturonic acid pathway and the MIOX and GalDH genes. Analysis of the promoters of AsA metabolism genes revealed a number of cis-acting elements related to developmental signals, but their functionalities remain to be investigated., This work was supported by research grants FP7-PEOPLE-2011-CIG-2011-303652 (Marie Curie Actions, European Union), Proyectos de I+D para Grupos de Investigación Emergentes GV/2012/036 (Generalitat Valenciana, Spain). The financial support of the projects AGL2015-70218 (Ministerio Economía y Competitividad, Spain) and PROMETEOII/2014/027 (Generalitat Valenciana) are also acknowledged. Enriqueta Alós was recipient of a JAE-Doc (CSIC) post-doctoral contract that was co-funded by Fondo Social Europeo de Desarrollo Regional (FEDER).

In this paper we present a method for estimating the height of the nocturnal stable boundary layer by using lidar measurements and a single radiosonde for unambiguous initial guess. The method relies on the correlation between aerosol stratifications in the stable boundary layer and minimum variance levels in the attenuated backscatter profile. The method is based on calculating either temporal or spatial variance vertical profiles of the attenuated backscatter and threshold-limited decision. A radiosonde temperature-based estimation is used to provide an initial guess if several minimum variance regions are detected. Two study cases using ceilometer data are shown. Comparison with temperature-based estimations from a collocated microwave radiometer have been used for validation. The method can be useful for estimating the stable boundary layer height in sites with a ceilometer but without any available temperature profiler. This work was supported via Spanish Government–European Regional Development Funds project PGC2018-094132-B-I00 and EU H2020 ACTRIS-IMP (GA 871115). CommSensLab is a María-de-Maeztu Unit of Excellence funded by the Agencia Estatal de Investigación, Spain. The work of M.P. Araujo da Silva was supported by the Spanish Ministry of Science, Innovation and Universities under Grant PRE2018-086054. Data were provided by Jülich Observatory for Cloud Evolution (JOYCE-CF), a core facility funded by Deutsche Forschungsgemeinschaft via grant DFG LO 901/7-1.

Durante el período de Aislamiento Social Preventivo y Obligatorio, en el contexto de pandemia por Covid-19, el teletrabajo en el hogar emergió como una solución válida para dar continuidad al habitual funcionamiento del sistema productivo ante la imposibilidad de traslado y de trabajo en forma presencial. El crecimiento exponencial de esta modalidad de trabajo, sumado al cierre de las instituciones educativas y de cuidados, acentuó la división sexual del trabajo no remunerado. Al calor de estas transformaciones se llevó al Congreso el debate por la Ley de Teletrabajo, que fue sancionada durante el año 2020. Entre las regulaciones que incorpora la ley, nos interesan particularmente las vinculadas a los sindicatos, en tanto nuevas problemáticas que surgen de esta modalidad laboral en el marco de los debates dados en torno al trabajo no remunerado. El objetivo central de este artículo es esgrimir los desafíos que enfrentarán las organizaciones sindicales frente a la reciente Ley de Teletrabajo, fundamentalmente en materia de género. Para ello recurriremos a la revisión bibliográfica, que nos permitirá problematizar la modalidad de teletrabajo en relación con el trabajo doméstico y de cuidados no remunerado y realizar un análisis crítico del articulado de la ley.

The purpose of this study was to evaluate the specific contribution of carotenoids and vitamin C to the lipophilic and hydrophilic antioxidant capacity, respectively, of the pulp of citrus fruits using the genetic diversity in pigmentation and in the carotenoid complement. To this end, six citrus varieties were selected: two mandarins, Clemenules (Citrus clementina) and Nadorcott (C. reticulata); two grapefruits (C. paradisi), Marsh and Star Ruby; and two sweet oranges (C. sinensis), Valencia late and Valencia Ruby. Total carotenoid content and composition in the pulp of fruits were very different, in relation to their color singularities. Valencia Ruby and Nadorcott had the highest carotenoid content, accumulating the former large amounts of linear carotenes (phytoene, phytofluene, and lycopene) and Nadorcott of β-cryptoxanthin. Orange fruits contained the highest amount of vitamin C while in Nadorcott mandarin it was substantially lower. Analysis of antioxidant capacity, evaluated by 2,2’-azino-di-(3-ethylbenzthiazoline sulfonate) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays, in the pulp of the different fruit varieties indicated a high and positive correlation between vitamin C content and hydrophilic antioxidant capacity. Nevertheless, a weak correlation was observed between carotenoids content and lipophilic antioxidant capacity in the pulp extracts assayed by ABTS. Overall, vitamin C in the pulp of citrus fruit had an important contribution to the hydrophilic antioxidant capacity, whereas that of carotenoids to lipophilic antioxidant capacity was very variable, being the highest that of Valencia Ruby orange, with large concentrations of lycopene and phytoene, followed by Nadorcott mandarin, with high β-cryptoxanthin content., JZ is the recipient of a FDEGENT/2018/007 predoctoral scholarship from Generalitat Valenciana (Spain) and FR is the recipient of a predoctoral scholarship from the Agencia Nacional de Investigación e Innovación (ANII, Uruguay). This work was supported by a research grants AGL2015-70218 and RTI2018-095131-B-I00 of the Ministry of Science, Innovation and Universities (Spanish Government). Authors are members of the CaRed Excellence Network (BIO2015-71703-REDT and BIO2017-90877-REDT) and European COST Action CA15113 EUROCAROTEN.

Andean Potato is a major crop for farmers in the Andes and represents an important gene pool for potato improvement. We present the population structure and genetic diversity of 88 Andigena accessions collected in Northwestern Argentina based on functional markers (25 SSR) distributed along 12 chromosomes. Polymorphic information content ranged from 0.40 to 0.87. A Bayesian approach, a Principal Coordinate Analysis and a Cluster Analysis revealed the presence of: I) a major group containing most of the Andean accessions and II) a smaller group including the out-group cv. Spunta and the sequenced genotype DM. Several group specific alleles were detected. AMOVA showed that 81% of the variability was within each group. Eleven of the SSRs analyzed are linked or within genes reported to regulate traits of nutritional and industrial interest. Additionally, the allelic variant of a photoperiod dependent tuberization regulator gene, StCDF1.2, was exclusively detected in accessions clustered in group II. Fil: Monte, Marcelo Nicolás. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Rey, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Carboni, Martín Federico. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Castellote, Martín Alfredo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Sucar, Sofía. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Norero, Natalia Sigrid. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Colman, Silvana Lorena. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Massa, Gabriela Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Colavita, Mónica Liliana. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Feingold, Sergio. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina

Reseña del libro: Feminismos y sindicatos en Iberoamérica. Ciudad Autónoma de Buenos Aires: Consejo Latinoamericano de Ciencias Sociales (CLACSO) y Universidad Nacional José C. Paz (UNPAZ), 2020. Nora Goren y Vanesa Lorena Prieto (Ed.)

During postpartum, high-production dairy cows show a temporary period of insulin resistance, during which glucose uptake by peripheral tissues is reduced to prioritize milk production. However, this can further increase their negative energy balance by compromising liver function, especially in cows with excessive body condition score (BCS) and a pro-inflammatory state. Based on this, the aim of this study was to evaluate the hepatic expression of proteins of the insulin signaling pathway (PI3K) and of the cytokines TNFα, IL-6 and NF-κB, as well as the plasma concentrations of non-esterified fatty acids (NEFA), beta-hydroxybutyrate, glucose, triglycerides (TAG), insulin and insulin-like growth factor-1, insulin sensitivity indexes, and the hepatic content of TAG during the transition period in cows with different BCS. Sixteen Holstein cows were selected 14 days before the expecting calving date and classified into 2 groups: low BCS (LBCS) ≤ 3.25 (n = 9) and high BCS (HBCS) ≥ 3.5 (n = 7). Blood and liver samples were obtained 14 (±3) days before the expected calving date and 4 (±3), 14 (±3) and 28 (±3) days after calving. The concentration of NEFA was higher in the HBCS group than in the LBCS group. Glucose concentration showed an interaction effect, with a greater concentration on day 28 in HBCS. Insulin concentration showed no changes. While the pAkt/total Akt ratio was lower in the HBCS group, the TNFα protein expression was higher only on day 4 postcalving in the HBCS group. In agreement with these results, the insulin sensitivity indexes RQUICKI and RQUICKIBHBA were lower in the HCBS group. The results suggest an insulin resistance and a pro-inflammatory state in the liver of cows with HBCS.

This work aimed to provide data about the molecules involved in induced resistance mediated by potassium phosphite (KPhi) in potato. To achieve these, we performed microarray analysis to determine the changes in gene expression after KPhi treatment,and their abundance after Phytophthora infestans(P.i.)infection.Our results showed the differential expression of 172 genes after KPhi treatment,and 22 KPhi primed genes after P.i. challenge, mainly involved insignal transduction and defense reactions,among others.Additionally,to identify common cis-elements involved in their regulation, we analyzed in-silico KPhi responsive genes promoters. Interestingly relevant phytohormone and stress-responsivecis-elements were present. Fil: Feldman, Mariana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina Fil: Guzzo, Maria Carla. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina Fil: Machinandiarena, Milagros Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina Fil: Rey Burusco, María Florencia. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: Beligni, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina Fil: Di Rienzo, Julio Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigación y Estudios de Matemática. Universidad Nacional de Córdoba. Centro de Investigación y Estudios de Matemática; Argentina Fil: Castellote, Martín Alfredo. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: Daleo, Gustavo Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina Fil: Andreu, Adriana Balbina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina

Ovulation is considered an inflammatory, cytokine-mediated event.Cytokines, which are recognized as growth factors with immunoregulatory properties, are involved in many cellular processes at the ovarian level. In this sense, cytokines affect fertility and are involved in the development of different ovarian disorders such as bovine cystic ovarian disease (COD). Because it has been previously demonstrated that ovarian cells represent both sources and targets of cytokines, the aim of this study was to examine the expression of several cytokines, including IL-1β, IL-1RA, IL-1RI, IL-1RII, IL-4 and IL-8, in ovarian follicular structures from cows with spontaneous COD. The protein expression of these cytokines was evaluated by immunohistochemistry. Additionally, IL-1β, IL-4 and IL-8concentrations in follicular fluid (FF) and serumwere determined by enzyme-linked immunosorbent assay (ELISA). In granulosa and theca cells, IL-1RI, IL-1RII, IL-1RA and IL-4 expression levels were higher in cystic follicles than in the control dominant follicles.The serumand FF concentrations of IL-1β and IL-4 showed no differences between groups, whereas IL-8 concentration was detected only in FF of cysts from cows with COD.The FF and serumconcentrations of IL-1β and IL-8 showed no significant differences, whereas IL-4 concentration was higher in FF than in serum in both the control and COD groups. These results evidenced an altered expression of cytokines in ovaries of cows with COD that could contribute to the pathogenesis of this disease. Fil: Stassi, Antonela Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Gareis, Natalia Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Marelli, Belkis Ester. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Matiller, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Leiva, Cristian Jesus Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Baravalle, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Intracellular lipid-binding proteins (iLBPs) of the fatty acid-binding protein (FABP) family of animals transport, mainly fatty acids or retinoids, are confined to the cytosol and have highly similar 3D structures. In contrast, nematodes possess fatty acid-binding proteins (nemFABPs) that are secreted into the perivitelline fluid surrounding their developing embryos. We report structures of As-p18, a nemFABP of the large intestinal roundworm Ascaris suum, with ligand bound, determined using X-ray crystallography and nuclear magnetic resonance spectroscopy. In common with other FABPs, As-p18 comprises a ten β-strand barrel capped by two short α-helices, with the carboxylate head group of oleate tethered in the interior of the protein. However, As-p18 exhibits two distinctive longer loops amongst β-strands not previously seen in a FABP. One of these is adjacent to the presumed ligand entry portal, so it may help to target the protein for efficient loading or unloading of ligand. The second, larger loop is at the opposite end of the molecule and has no equivalent in any iLBP structure yet determined. As-p18 preferentially binds a single 18-carbon fatty acid lig- and in its central cavity but in an orientation that differs from iLBPs. The unusual structural features of nemFABPs may relate to resourcing of developing embryos of nematodes. Fil: Ibáñez, Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentina Fil: Rey Burusco, M. Florencia. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; Argentina Fil: Gabrielsen, Mads. University of Glasgow; Reino Unido Fil: Franchini, Gisela Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentina Fil: Riboldi Tunnicliffe, Alan. Australian Synchrotron; Australia Fil: Roe, Andrew J.. University of Glasgow; Reino Unido Fil: Griffiths, Kate. University of Glasgow; Reino Unido Fil: Cooper, Alan. University of Glasgow; Reino Unido Fil: Córsico, Betina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentina Fil: Kennedy, Malcolm W.. University of Glasgow; Reino Unido Fil: Smith, Brian O.. University of Glasgow; Reino Unido

Cattle undergo numerous environmental and management stressors that reduce fertility and affect ovulation. The extracellular matrix of the follicle wall can be altered by matrix metalloproteinases (MMPs), the activities of which are regulated by interleukins and tissue-specific inhibitors of metalloproteinases (TIMPs), especially during ovulation. The aims of the present study were to: (1) evaluate changes in the hormone milieu, the localisation and activity of MMP2 and MMP9 and the localisation of MMP14, TIMP1 and TIMP2 in response to adrenocorticotrophic hormone (ACTH) during the preovulatory period in cows; and (2) determine the direct effects of ACTH on the mRNA expression of MMP2 and MMP9 in the cultured follicle wall of bovine ovaries obtained from an abattoir. 100IU ACTH was administered during pro-oestrus every 12h until ovariectomy, which was performed before ovulation. Cortisol concentrations in the plasma and follicular fluid (FF) of preovulatory follicles were higher in ACTH-treated than control cows. Progesterone presented subluteal concentrations in plasma of ACTH-treated cows (P

We aimed to study the protein and gene expression of some hepatic enzymes of lipid metabolism along with plasma biomarkers in grazing dairy cattle during the transition period. Blood and liver biopsies from a group of eight multiparous cows were sampled at −28, −14, +4, +14, +28 and +56 days relative to parturition. Peak concentrations of NEFA and beta-hydroxybutyric acid with high triacylglycerol content in the liver were recorded on day 4 postpartum. Consistent with blood biomarkers, the gene expression of carnitine palmitoyltransferase 1A (CPT1A) and acyl-CoA oxidase 1 (ACOX1) increased, whereas that of diacylglycerol O-acyltransferase 1 (DGAT1) decreased. Nevertheless, CPT1A protein expression did not change during all the period evaluated and ACOX1 protein expression increased on day 56 postpartum. In addition, the protein expression of peroxisome proliferator-activated receptor alpha (PPAR-alpha) increased on day 28 postpartum. On the other hand, DGAT1 protein expression decreased on day 14 postpartum. As expected, the expression of genes associated with fatty acid oxidation increased on the first days postpartum but, notably, protein expression was highest after transition. Since most infectious diseases and metabolic disorders in dairy cattle occur particularly on the first days postpartum, it is not so clear whether an increase in the oxidation capacity of the liver at that time could help to prevent disease and improve dairy production. The valuable results about protein expression of enzymes involved in liver lipid metabolism could help to better characterize the metabolism of dairy cattle during the transition period. Fil: Angeli, Emmanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Trionfini, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina Fil: Gareis, Natalia Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Matiller, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Huber, Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Hein, Gustavo Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Cystic ovaries (CO) characterize a disorder frequently found in dairy cattle. However, despite the contributions by several researchers, the mechanism that leads to ovulatory failure has not yet been completely elucidated. Thus, the aim of this study was to examine the mRNA expression of bovine vascular endothelial growth factor (VEGFA)-164, VEGFA 164b and VEGF receptors (VEGFR1 and VEGFR2) by real-time PCR and protein expression by immunohistochemistry, immunofluorescence and western blot in follicular fluid from dairy cows with spontaneous CO and in an experimental model of follicular persistence induced by prolonged treatment with progesterone. Results showed that both VEGFA isoforms and receptors were coexpressed in granulosa and theca interna cells and in follicular fluid of ovaries from all the groups evaluated. VEGFA-164, VEGFA-164b and VEGFR2 protein expression was higher in theca cells of persistent follicles from group P0 (expected time of ovulation) than in those from dominant follicles (as reference structure) from the control group (p < 0.05). Also, VEGFA-164 expression was higher in theca cells of cysts than in those of dominant follicles of the control group (p < 0.05). In follicular fluid, VEGFA-164 expression was higher in persistent follicles from group P5 (5 days of follicular persistence) than in the control, P0 and P15 groups, and higher in cysts than in dominant follicles from the control group (p < 0.05). This study provides evidence of an altered expression of VEGFA-164, VEGFA-164b and VEGFR2 during the formation of persistent follicles and cysts in cows. Together, these results evidence that early development of CO in cows is concurrent with an altered expression of these growth factors and that these alterations may contribute to the follicular persistence, angiogenic dysregulation and ovulatory failure found in cows with follicular cysts. Fil: Stassi, Antonela Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Gasser, Fátima Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Velázquez, Melisa María del Luján. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Belotti, Eduardo Matías. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Gareis, Natalia Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Baravalle, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Folliculogenesis and ovulation are regulated by gonadotrophins and other factors such as Insulin like growth factor 1 (IGF1) and leptin. In various species the presence of IGF1 receptor (IGF1R) and leptin receptor (ObR) has been detected in the ovary, but not in the alpaca. Thus, the aim of the present study was to evaluate the presence of these receptors in this tissue and analyze if the presence of these receptors in the ovary is related to the presence of a corpus luteum (CL) and if abundances, as determined by immunostaining intensity vary with follicle size. The IGF1R and ObR were identified in primary and secondary follicles, granulosa and theca interna cells of tertiary follicles and in CL. There were greater abundances of IGF1R in granulosa cells of tertiary follicles of ovaries without compared with those with CL. In both groups, the immunostaining of granulosa cells was greater than in theca interna cells. The abundance of ObR was greater in primary and secondary follicles, and theca interna cells of tertiary follicles in ovaries with than those without CL. Immunostaining of granulosa cells was greater than theca interna cells only in ovaries without CL. There were no differences in the abundance of ObR and IGF1R between primary and secondary follicles and granulosa cells of tertiary follicles, neither in ovaries with or without CL. The abundance of IGF1R was not correlated with abundance of ObR neither in ovaries with or without CL. These results indicate a possible role for IGF and leptin in ovarian function. Furthermore, these receptors could be regulated by ovarian steroid hormones because abundance of these receptors in ovaries varies depending on whether there is a CL present in the ovary. Fil: Gallelli, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina Fil: Bianchi, Carolina Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina Fil: Lombardo, Daniel Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rodríguez, Fernanda Mariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Castillo, Victor Alejandro. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina Fil: Miragaya, Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Ovarian cystic follicles are the sign of important causes of reproductive failure in numerous species. In this review, some morphological, endocrinological and clinical aspects of cystic follicles in women, cows, mares, sows and bitches are discussed. Follicular cysts are the consequence of the failure of a mature follicle to ovulate at the appointed time of ovulation in the estrous cycle. Although the etiology of follicular cysts remains unknown, this review examines the evidence about the role of endocrine signaling systems in the specific disease or syndrome in each of the species mentioned above. This review also describes, the changes in the pathways of endocrine mechanisms that would trigger disturbances in the intraovarian component underlying the aberrant persistence of follicular cysts. The knowledge of the morphological and endocrinological nature of cystic follicles in different species can provide relevant information to better understand specific diseases when it is integrally analyzed from the comparative medicine viewpoint. Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Díaz, Pablo Uriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Hein, Gustavo Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Marelli, Belkis Ester. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rodríguez, Fernanda Mariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Stassi, Antonela Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

La enfermedad quística ovárica (COD) es una de las principales causas de falla reproductiva y de infertilidad, y constituye uno de los de los trastornos reproductivos más frecuentes en vacas lecheras de alta producción. El desarrollo de esta enfermedad está asociado a un desequilibrio en el eje hipotalámico-hipofisario-gonadal en el cual factores endocrinos, entre ellos el sistema de factores de crecimiento análogos a insulina (IGF), participan en el desarrollo folicular, diferenciación celular y la secreción de hormonas ováricas. Considerando al sistema IGF como un importante regulador del crecimiento y funcionalidad folicular, evaluamos la participación de componentes del sistema IGF en el desarrollo de la COD. Las alteraciones en diferentes componentes del sistema IGF que podrían afectar al normal funcionamiento del ovario participando en el desarrollo de la COD en bovinos Fil: Rodríguez, Fernanda Mariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Ambient temperatures that result in body temperatures beyond those of the thermo-neutral zone for dairy cattle can lead to reduced reproductive efficiencies that have negative effects on economic and productive efficiencies of dairy farms. In addition, in pregnant cows, ambient temperature-induced heat stress leads to modifications in the epigenome of the developing embryo, which, in turn, could lead to phenotypic variations in the sexually mature animal and its offspring. In the mammalian response to stress, adrenocorticotropic hormone stimulates the synthesis and secretion of glucocorticoids, which may have detrimental effects on the hypothalamic-pituitary-gonadal axis and the female estrous cycle. The aim of this review is to describe the effects of ambient heat stress on the reproductive system of dairy cattle and its potential trans-generational effects. There are many heat stress occurrences in dairy cattle during a large portion of the year in many countries and there is an increase in incidence with the onset of global warming. These heat stress conditions make it possible that the embryo/fetus of cows may be affected when heat stress conditions prevail in ways that there is impaired fertility of the sexually mature cows that develop from these embryos/fetuses. This is the outcome because of molecular changes in ovarian glucocorticoid response caused by epigenetic modifications established during fetal development.

Resumen: En la cáscara de mandarina Satsuma ‘Okitsu’ se detectó en Uruguay un manchado durante la postcosecha, ocasionando en algunos años importantes pérdidas. En este trabajo se estudió el daño a nivel micro y macroscópico, su asociación con factores ambientales previos a la cosecha y el efecto del desverdizado y encerado de frutos sobre su incidencia y severidad. A partir de los 15 días de almacenamiento, se visualizó entre las glándulas de aceite una zona irregular de color pardo. Luego de 15 días de mostrador, en las zonas con síntomas más severos, se observó colapso de células subepidérmicas, avanzando hacia el parénquima subyacente y en algunos casos hacia la epidermis, sin afectar a la cutícula. En los dos primeros años evaluados, la incidencia de frutos manchados fue de 17 % y 7 % respectivamente, mientras que en los dos últimos no se registraron frutos afectados. La mayor incidencia se asoció a un período de mayor temperatura máxima, menor humedad relativa y menores precipitaciones durante los 10 días previos a la cosecha. El desverdizado no afectó la incidencia de frutos manchados a los 30 días de almacenamiento + 15 de vida de mostrador, mientras que el encerado de frutos la disminuyó en más de un 50 %. Los resultados sugieren que las condiciones ambientales precosecha favorables a la deshidratación de frutos y/o las condiciones postcosecha que no logren evitar la pérdida de agua, promueven el manchado de los frutos de mandarina Satsuma ‘Okitsu’. Summary: During postharvest of Satsuma ‘Okitsu’ mandarin it has been detected in Uruguay a rind breakdown, which some years causes important losses. In this work, we studied microscopic and macroscopic damage, its association to environmental preharvest conditions, and the effect of postharvest fruit degreening and waxing on its incidence and severity. After 15 days of storage, irregular brownish areas boarding oil glands started to appear in the peel. After 15 days of shelf life, in areas with severe symptoms, a depression and cell collapse could be detected at sub-epidermis, advancing to subjacent parenchyma and occasionally to the epidermis, but the cuticle was not affected. In the former two years, fruit rind breakdown incidence was respectively 17 % and 7 %, while no damaged fruits were detected in the following two years. The highest rind breakdown incidence was associated to higher maximum temperature, lower relative humidity, and less precipitation during the last 10 days before harvest. Fruit degreening did not affect rind breakdown incidence after 30 days of storage + 15 days of shelf life, while fruit waxing diminished it more than 50 %. Results suggest that preharvest environmental conditions that promote fruit dehydration and/or postharvest conditions that fail to prevent fruit water loss, promote this ‘Okitsu’ Satsuma mandarin rind breakdown.

Cystic ovarian disease (COD) represents an important cause of infertility in dairy cattle and is associated with multiple physiological disorders. Steroidogenesis, which is necessary to ensure normal ovarian functions, involves multiple enzymatic pathways coordinated by insulin and other proteins. We have previously shown that cows with COD have an altered insulin response. Therefore, in the present study, we evaluated further alterations in intermediates downstream of the PI3K pathway and pathways mediated by ERK as critical signals for the expression of steroidogenic enzymes in the ovaries of control cows and cows with spontaneous COD. To this end, we evaluated the gene and protein expression of pan-AKT, mTOR, ERK1/2, and steroidogenic enzymes by real-time PCR and immunohistochemistry. Steroid hormone concentrations were assessed at systemic and intrafollicular level. Results showed altered expression of intermediate molecules of the insulin signaling pathway, whose action might modify the synthetic pathway of steroidogenic hormones. Similarly, the expression of steroidogenic enzymes and the concentration of progesterone in serum and follicular fluid were altered. These alterations support the hypothesis that systemic factors contribute to the development and/or maintenance of COD, and that metabolic hormones within follicles such as insulin exert determinant effects on ovarian functionality in cows with COD. Fil: Gareis, Natalia Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Huber, Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Hein, Gustavo Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rodríguez, Fernanda Mariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Angeli, Emmanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

In ovarian granulosa cells, follicle-stimulating hormone (FSH) regulates the proliferation and differentiation events required for follicular growth and oocyte maturation. FSH actions are mediated exclusively through the FSH receptor (FSHR). In cattle, the FSHR gene expression pattern during folliculogenesis and the implications of this receptor in reproductive disorders have been extensively studied. However, the limited availability of specific antibodies against bovine FSHR has restricted FSHR protein analysis. In the present study, we developed an anti-FSHR polyclonal serum by using a 14-kDa peptide conjugated to maltose binding protein. The antiserum obtained was characterized by western blot of protein extracts from bovine follicles, BGC-1 cells and primary cultures of granulosa cells stimulated with testosterone. Also, the blocking effect of serum on estradiol secretion and cell viability after gonadotropin stimulus was characterized in a functional in vitro assay. A 76-kDa protein, consistent with the predicted molecular size of full-length FSHR, was detected in ovarian tissue. Besides, two immunoreactive bands of 60-kDa and 30-kDa (only in cultured cells) were detected. These bands would be related to some of the isoforms of the receptor. Therefore, immunohistochemical assays allowed detecting FSHR in the cytoplasm of granulosa cells and an increase in its expression as follicles progressed from primordial to large preantral follicles. These results suggest that the anti-FSHR serum here developed has good reactivity and specificity against the native FSHR. Therefore, this antiserum may serve as a valuable tool for future studies of the biological function of FSHR in physiological conditions as well as of the molecular mechanism and functional involvement of FSHR in reproductive disorders. Fil: Marelli, Belkis Ester. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Leiva, Cristian Jesus Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Flores Brun, Rocío B.. Universidad Nacional del Litoral; Argentina Fil: Ramírez, Cintia S.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina Fil: Failla, J.I.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina Fil: Matiller, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Amweg, Ayelen Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

High-producing dairy cows frequently suffer metabolic alterations that cause different diseases, which could decrease the reproductive efficiency of the herd. Among these reproductive disorders, cystic ovarian disease (COD) has been related to alterations in metabolites and hormonal factors such as insulin, adiponectin and leptin. The aim of this study was to determine the protein expression of adiponectin and some of its downstream targets in ovarian follicles of control cows and cows with clinical diagnosis of COD. We also analyzed some key metabolic sensors in plasma and follicular fluid from both groups. In follicular cysts, we detected higher protein expression of adiponectin receptor 2 (AdipoR2), 5′ adenosine monophosphate-activated protein kinase (AMPK), carnitine palmitoyl transferase 1 (CPT1) and acyl-coenzyme A oxidase 1 (ACOX1) relative to control antral follicles (p < 0.05). This was related to higher plasma adiponectin concentration in cows with COD than in control cows (p < 0.05). On the other hand, insulin concentrations showed an opposite pattern (p < 0.05). Furthermore, we found alterations in local and systemic concentrations of several metabolites. In this regard, in follicular fluid of cystic cows, the concentrations of non-esterified fatty acids and beta-hydroxybutyrate were higher (p < 0.05), whereas the concentrations of glucose and triacylglycerol were lower than in follicular fluid from control cows (p < 0.05). Besides, in both follicular fluid and plasma of cows with COD, the concentration of cholesterol was higher than in control animals (p < 0.05). These results evidence a local altered scenario of some metabolic sensors in cystic follicles, which could generate an adverse microenvironment for the resumption of ovarian activity, possibly causing the persistence of follicles and the recurrence of COD. Fil: Gareis, Natalia Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Angeli, Emmanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Huber, Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rodríguez, Fernanda Mariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Hein, Gustavo Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Anti-Müllerian hormone (AMH) is a homodimeric glycoprotein expressed exclusively in the gonads. This hormone is an important regulator of the early growth of follicles through inhibitory effects on the recruitment of primordial follicles into the pool of growing follicles and on granulosa cell proliferation. Cystic ovarian disease (COD) is an important disorder affecting the fertility of dairy cattle. In the present study, we evaluated the expression of AMH in granulosa cells and AMH secretion into follicular fluid in pre-ovulatory follicles from control cows, animals with spontaneously arising COD and during the development of the disease, at 5, 10 and 15 days of follicular persistence. To this end, after an oestrous synchronization protocol, low doses of progesterone was administered for 5, 10 and 15 days after the expected day of ovulation (day 0 of follicular persistence) in treated cows (groups P5, P10 and P15, respectively), using an intravaginal progesterone-releasing device. Results showed a decrease in the expression of AMH in granulosa cells throughout folliculogenesis (P

Fil: Matiller, Valentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Cs.veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina