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The aim of the study was to investigate the potential of a diet containing rapeseed meal fermented by Saccharomyces cerevisiae as a new sustainable feed to reduce transient intestinal inflammation, diarrhoea and oxidative stress in piglets after weaning. In this study, 16 male post-weaning piglets, with an initial weight of 9.04 ± 0.19 kg, were randomly allocated to two dietary treatments: control and 10% fermented rapeseed meal (FRSM) -- 8 pigs/treatment. The experiment lasted 21 days. At the end of the trial, the animals were slaughtered and samples of blood and segments of the jejunum, ileum and colon were collected for determination of plasma biochemical, inflammatory and oxidative stress parameters. Pig performance and diarrhoea incidence were also investigated. The results showed that the FRSM diet had no significant effect on piglet performance, weight and average daily weight gain, as well as plasma biochemical parameters. However, the number of piglets with diarrhoea was higher in the control group than in the group receiving the FRSM diet throughout the experimental period. Moreover, a decrease in TNF-α (P = 0.03) and IL-1ß (P < 0.05) cytokine levels was recorded in the colon and jejunum samples from the FRSM group. In addition, IL-8 and IL-6 concentrations were decreased (P = 0.0009 and P = 0.03, respectively) in the ileum of piglets fed FRSM, indicating the modulatory capacity of this feed in reducing weaning-associated intestinal inflammation. The FRSM diet also improved the antioxidant status and significantly reduced lipid peroxidation and thiobarbituric acid reactive substance levels in plasma (P = 0.022) and in the jejunum (P = 0.028) and colon (P = 0.003), suggesting the potential of fermented rapeseed meal to limit oxidative reactions. In conclusion, the present study showed that fermentation of rapeseed meal using S. cerevisiae enriched the nutrient composition and reduced the concentration of anti-nutrients (e.g. glucosinolates). Moreover, the addition of FRSM to diets of pigs after weaning improved their intestinal health status, indicating its beneficial effect. [ABSTRACT FROM AUTHOR]

Significant interest exists on the inclusion of oilseeds in dairy cows' diets to improve the polyunsaturated fatty acid (PUFA) profile of their milk due to PUFA's effects on human health. However, this strategy can affect milk protein fractions, compounds that play an important role in the dairy industry, and also affect consumers' health. The present work thus aimed to investigate the effect of rapeseed in dairy cows' diets on milk protein fractions such as caseins [as1-casein (as1-CN), as2-casein (as2-CN), ß-casein (ß-CN), k-casein (k-CN)], whey proteins [ß-lactoglobulin (ß-Lg), a-lactalbumin (a-La)], and minor proteins [lactoferrin (LF), bovine serum albumin (BSA), immunoglobulin G (IgG)]. The animals were randomly distributed in two groups, each fed with a control diet and a rapeseed-based diet, respectively. The protein fractions were determined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Results indicated that the inclusion of rapeseed in dairy cows' diets led to a significant decrease in total milk protein (from 3.73 ± 0.03 to 3.58 ± 0.03%, p < 0.05) and total casein content (from 2.96 ± 0.03 to 2.80 ± 0.03%, p < 0.05). Additionally, a significant decrease was also observed in the content of k-CN (from 32275 to 25883 A.U., p = 0.015), an important protein involved during milk coagulation; and a decrease tendency for ß-CN also occurred (from 44746 to 41441 A.U., p = 0.087), the high content of which is associated with some human diseases. Dietary rapeseed also decreased the ß-Lg content (from 61256 to 52694 A.U., p = 0.016), which is positively correlated with cheese quality. Other minor protein fractions such as BSA and IgG were also decreased by the rapeseed diet. [ABSTRACT FROM AUTHOR]

Objetivo. Este estudio es un paso preliminar de un programa nacional más amplio destinado a desarrollar una estrategia para la conservación "in situ"de la población de ovejas Cabeza Negra de Teleorman. En este trabajo se estimó el efecto de la β-lactoglobulina, caseína y prolactina en algunos rasgos cuantitativos y cualitativos de la leche en esta población de ovejas locales. Material y métodos. Metodología de PCR para genotipificación incluido CSN3 (A y B alelos) y PCR-RFLP para LGB (A y B alelos) y PRL (T y C alelos). Análisis y composición de la leche de ordeños repetidos se utilizaron para estimación el efecto del polimorfismo. Resultados. No se encontró asociación entre el polimorfismo y la leche rasgos CSN3. Efecto de LGB en los rasgos de producción era bastante constante. Genotipo AA obtenido mejores resultados que BB. Efecto marcador PRL mostró pequeñas diferencias que LGB. En cuanto a la leche grasa y proteína el genotipo AA para PRL tuvo un impacto positivo más pequeño que el genotipo AA para LGB. En cuanto a contenido de grasa y proteína, PRL mostró un efecto negativo para AA y positivo para BB genotipo. Conclusiones.La asociación positiva entre LGB y la producción de leche y la composición recomienda este gen candidato como marcador para un futuro programa de MAS. Aunque gen PRL también se asocia con un incrementoen la cantidad de leche, la respuesta inversa sobre composición de la leche debe ser considerado en la estrategia de MAS. Nuestro estudio demostró que los marcadores tanto LGB y PRL podrían venir a ser utilizados en MAS en la industria rumanade cría deovejas lecheras. [ABSTRACT FROM AUTHOR]

Objective. This study is a preliminary step of a larger national program aimed to develop a strategy for "in situ" preservation of Teleorman Black Head sheep population. In this paper we estimated the effect of β-lactoglobulin, casein and prolactin on some quantitative and qualitative milk traits in this local sheep population. Material and methods. Genotyping methodology included PCR for CSN3 (A and B alleles) and PCR-RFLP for LGB (A and B alleles) and PRL (T and C alleles), respectively. Repeated milking and milk composition analysis were used for the polymorphism effect estimation. Results. No association between CSN3 polymorphism and milk traits was found. Effect of LGB on production traits was quite constant. Genotype AA performed better than BB. PRL marker effect showed small differences than LGB. Concerning milk, fat and protein yield, AA genotype for PRL had a smaller positive impact than AA genotype for LGB. Regarding fat and protein content, PRL showed a negative effect for AA and positive for BB genotype, respectively. Conclusions. Positive association between LGB and milk yield and composition recommend this candidate gene like marker for a future MAS program. Although PRL gene is also associated with an increased milk quantity, inverse response over milk composition must be considered in MAS strategy. Our study demonstrated that both LGB and PRL markers could became an advent of MAS utilization in Romanian dairy sheep breeding industry. [ABSTRACT FROM AUTHOR]

Objective. This study is a preliminary step of a larger national program aimed to develop a strategy for “in situ” preservation of Teleorman Black Head sheep population. In this paper we estimated the effect of β-lactoglobulin, casein and prolactin on some quantitative and qualitative milk traits in this local sheep population. Material and methods. Genotyping methodology included PCR for CSN3 (A and B alleles) and PCR-RFLP for LGB (A and B alleles) and PRL (T and C alleles), respectively. Repeated milking and milk composition analysis were used for the polymorphism effect estimation. Results. No association between CSN3 polymorphism and milk traits was found. Effect of LGB on production traits was quite constant. Genotype AA performed better than BB. PRL marker effect showed small differences than LGB. Concerning milk, fat and protein yield, AA genotype for PRL had a smaller positive impact than AA genotype for LGB. Regarding fat and protein content, PRL showed a negative effect for AA and positive for BB genotype, respectively. Conclusions. Positive association between LGB and milk yield and composition recommend this candidate gene like marker for a future MAS program. Although PRL gene is also associated with an increased milk quantity, inverse response over milk composition must be considered in MAS strategy. Our study demonstrated that both LGB and PRL markers could became an advent of MAS utilization in Romanian dairy sheep breeding industry.

Objective. Estimate the genetic parameters for milk traits in a Romanian local sheep population Teleorman Black Head. Material and methods. Records of 262 sheep belonging to 17 rams and 139 ewes were used in the study. The following traits were investigated: milk yield, fat yield, protein yield, fat percentage and protein percentage. The genetic parameters were estimated using the Restricted Maximum Likelihood method, with a model including maternal effects. Results. The results from our study revealed that direct heritability estimates were moderate for milk yield (0.449), fat yield (0.442), protein yield (0.386) while for protein percentage (0.708) and fat percentage (0.924) were high. The high direct and maternal genetic correlation was between milk yield and protein yield (0.979, 0.973) and between protein yield and fat yield (0.952, 0.913) while the phenotypic correlation between the milk yield and fat yield (0.968), the milk yield and protein yield (0.967), fat yield and protein yield (0.936) was high and positive. Conclusions. The genetic parameters are important in selection program on this breed for genetic improvement.

First colostrum yield and constituents as well as milk yield during established lactation vary considerably among mammary quarters in dairy cows. However, data on the development of milk yield, IgG concentration, and their distribution per quarter within cows during the first milkings after calving are scarce. We analyzed milk production and IgG concentration at the individual quarter level in 29 multiparous Holstein cows during the first 5 milkings after calving. Cow- and calf-related factors (time interval between calving and first milking, parity number, previous lactation yield, gestation length, dry period length, sex, and birth weight of the calf) potentially affecting first colostrum quality and quantity were assessed. Milking of first colostrum was carried out between 30 and 180 min after parturition. Further milkings were performed twice daily. Quarter milk yield varied between 0.1 and 5.5 kg at the first milking and between 1.4 and 5.1 kg at the fifth milking relative to parturition. Quarter IgG concentration ranged between 18.8 and 106.0 mg/mL at the first milking and between 0.8 and 46.1 mg/mL at the fifth milking. Distribution of milk yield and IgG concentration among quarters was not entirely repeatable during the first 5 successive milkings after parturition; that is, the ranking of quarters changed (intraclass correlation coefficients for quarter milk yield and IgG concentration: 0.64 and 0.79, respectively). The average hourly milk production increased in all quarters, ranging from 0.02 to 0.26 kg/h between the first 2 milkings up to 0.11 to 0.45 kg/h between the fourth and fifth milkings. First colostrum yield was not affected by any of the evaluated cow- and calf-related factors. Quarter colostrum IgG concentration was higher in cows with a higher previous lactation yield, whereas a lower colostrum IgG content was observed in cows with a longer gestation period and consequently heavier calves. In conclusion, milk yield and IgG concentration of individual quarters varied considerably, and their distribution among quarters within cows was moderately repeatable in consecutive milkings and changed partially over time. The decline of IgG concentration was independent of the concomitant increase in milk secretion, with changes occurring at different rates in individual quarters. Our results confirm the independence of the single mammary quarters at the onset of lactation despite an identical exposure to endocrine stimuli., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

It was suggested that the immune system plays an important role at least in the amplification of the main elements in systemic sclerosis (SSc), an autoimmune disease with an incompletely elucidated pathogenesis. Elucidation of the mechanisms involved in the interaction between T and B cells, major players of the immune system, could contribute to a better understanding of some of clinical and pathological manifestations of SSc. Recently, abnormalities in Semaphorin 4D (Sema4D/CD100) or CD72, two contrareceptors involved in T and B cells cooperation, were associated with autoimmunity. Therefore, we investigated CD100 and CD72 expression level on T and B cells in attempting to establish their role in SSc pathogenesis. The results revealed augmented percentages of CD100(high) T and B cells, significantly increased expression of CD100 on CD4(+) T cells and frequently detectable levels of soluble CD100 in SSc patient sera compared to healthy donors. In SSc, CD100 dysregulations were associated with anti-Scl70 antibodies production, disease type, thickening of skin, disease duration, or with active inflammation processes. In consequence, dysregulations in CD100 expression and release could play a role in SSc development and/or maintenance.

PI3K/Akt/mTOR signaling pathway plays an important role in cellular proliferation and growth signaling. It was demonstrated that murine models presenting activated PI3K/Akt/mTOR signaling pathway in lymphocytes develop features of systemic autoimmunity, linking this pathway to autoimmune diseases. Therefore, the goal of our study was to analyze this signaling axis in Systemic Lupus Erythematosus (SLE), the prototype of systemic autoimmune diseases, focusing on Akt and p70S6k, two components of this pathway. Our results demonstrated that both expression and phosphorylation levels of Akt are more increased in SLE than in healthy donors (HDs) CD4+ T cells suggesting an up-regulation of PI3K and mTOR activities. This result was also suggested when p70S6k, one of mTOR substrate, was evaluated. Indeed, in SLE CD4+ T cells an enhancement of p70S6k activity, in direct correlation with its expression level, was found. Since p27kip1, an inhibitor of cell cycle progression, is one of the Akt substrates, we analyzed its expression level in relationship with cell cycle progression and apoptosis. The results demonstrated that p27kip1 expression level was significantly decreased in SLE than in HDs CD4+ T cells. In SLE p27kip1 level was inversely correlated with the percentage of peripheral lymphocytes in apoptosis and in S phase of the cell cycle. Therefore, the increased activity of PI3K/Akt/mTOR signaling pathway and, as a result, the drop of p27kip1 levels observed in CD4+ T cells isolated from SLE patients might explain the accumulation of SLE lymphocytes in S and G2/M cell cycle phases where they undergo apoptosis.

Immunologic abnormalities observed in Systemic Sclerosis (SSc) patients consist of chronic mononuclear cell infiltration of affected tissues, dysregulation of lymphokine and growth factor production, and autoantibodies production. Expansion of CD4+T cells within the tissue seems to involve their activation that precedes this process. Therefore, CD4+T cells activation, as an early immune event, appears to be an important process in the development and maintaining of SSc. In SSc the disturbance of peripheral tolerance mechanisms could be also responsible for CD4+T cells activation. Consequently, we reevaluated CD4+T cells positive for CD25, GITR, CTLA-4, CD45RO, or Foxp3 in SSc patients, by comparison with healthy donors (HDs), and in correlation with clinical features of the disease. Our results reargued for activation of peripheral blood CD4+T cells in SSc patients. Thus, increased percentages of CD25+ and GITR+ CD4+T cells were found in SSc patients by comparison with HDs. Direct correlation between the percentage of GITR+CD4+T cells and disease activity recommended these cells as a good candidate for disease progression. In SSc patients, the negative regulators of T cells activation are also affected. Thus, CTLA-4+ and Foxp3+ CD4+T cell percentages were significantly reduced in SSc patients when compared to HDs. Indirect correlation between the percentage of CD152+CD4+T cells and autoantibodies (aScl70) presence or disease type highlighted the role of these cells in the disturbance of peripheral tolerance. The absence of the direct correlation between CD152+CD4+T cells and CD45RO+CD4+T cells, correlation observed only in HDs, raised the hypothesis that in SSc patients, memory T cells can be easily activated, and by consequence, they can enter within affected tissues. These data reconfirm the activation state of SSc CD4+T cells and point out some abnormalities in peripheral tolerance mechanisms that can contribute to SSc pathogeny.

The aim of our study was to investigate and characterize regulatory T cells (Treg) in peripheral blood of patients with connective tissue diseases (Systemic lupus erythematosus, systemic sclerosis, Sjögren's syndrome, poly- and dermatomyositis) as compared with blood from healthy controls. Treg cells were quantified and phenotypically characterized by flow cytometry while the expression level of Foxp3 mRNA was evaluated by real time PCR. A reduced percentage of peripheral blood Treg cells was found in patients than in controls, irrespective of the type of connective tissue disease. Treg cells, especially those expressing one of the phenotypical markers, seemed to differ not only between patients and healthy controls but also among types of diseases. Additionally, the presence of autoantibodies as well as disease activity appeared to be correlated with particular Treg cell populations, especially those expressing one of the examined phenotypical markers. Correlations with therapy suggested that glucocorticoids plus antimalarial or other immunosuppressor drugs diminished the percentage of Treg cells, especially of those with memory phenotype. These findings indicated dysregulations at the level of Treg cells and suggested an involvement of these cells in the pathology of connective tissue diseases. Moreover, our data are in agreement with the suggestion that Treg cells could be therapeutic targets for some autoimmune diseases.

The cellular and molecular mechanisms involved in many abnormalities described in Systemic Lupus Erythematosus (SLE) are still unclear. Some of these abnormalities referred to the hyperactivation of T lymphocytes and the enhanced secretion of MMP-9 by peripheral blood mononuclear cells (PBMCs). Therefore, in this paper we investigated the potential role of CD147 molecule in these abnormalities. Our results demonstrated that CD147 molecule is overexpressed on CD3+T lymphocytes from SLE patients when compared with CD3+T lymphocytes from healthy donors. Monoclonal anti-CD147 antibodies, MEM-M6/1 clone, were able to inhibit protein tyrosine phosphorylation only in CD3 x CD28 costimulated T lymphocytes from SLE patients. However, this monoclonal antibody was unable to inhibit the enhanced activity of MMP-9 secreted by SLE PBMCs.

In this study, the levels of matrix metalloproteinases MMP-2 and MMP-9 were simultaneously analyzed with the levels of their tissue natural inhibitors TIMP-1 and TIMP-2 in sera of patients with breast tumors. At the same time, the activity of these two matrix metalloproteinases was evaluated. The decrease of TIMP-2 level in sera from patients with breast cancer as well as an imbalance between MMP-2 and TIMP-2 in neoplasic processes were found. The serum levels of MMP-2, MMP-9 and TIMP-1 were comparable between the patients with breast cancer and benign tumors. These experimental studied parameters were found to correlate with some of clinicopathological disease variables (TNM or pTNM staging system, tumor size and node invasion) suggesting their potential value for diagnosis and prognosis of breast cancer. Matrix metalloproteinases or their natural inhibitors and tumor markers (CA15.3 and CEA) not correlated between but, each of them correlated with another clinicopathological disease variable, suggesting their usefulness in the evaluation.