Management during the dry period and its effect on hepatic and adipose tissue molecular biomarkers of metabolism and health in grazing dairy cows

A successful transition into lactation determines optimum production, reproduction, and health. The peripartum period is characterized by an inflammatory state that, if not controlled, could be detrimental to the cow. The first experiment examined hepatic and adipose gene expression in response to injections of a non-steroidal anti-inflammatory compound (Carprofen) on 1, 3, and 5 d postpartum. Results indicated that after calving both tissues respond to inflammation signals, underscoring its role in the normal homeorhetic adaptations to lactation. The second experiment investigated the effect of prepartal nutrition and its interaction with BCS on hepatic and adipose tissue transcriptome, and the liver one-carbon metabolism and transulfuration pathway. Cows were randomly allocated to one of four groups in a 2 × 2 factorial arrangement: 4.0 or 5.0 BCS prepartum (10-point scale) and dietary energy at 75 or 125% of estimated requirements during the close-up. Tissue biopsies were harvested at -1, 1 and 4 wk relative to parturition. The greater number of hepatic differentially expressed genes in BCS4 cows in response to increased prepartum feed allowance (1071 vs 310, over the entire transition period) indicated a greater responsiveness to prepartum nutrition than optimally-conditioned cows. Thus, overfeeding in late-pregnancy should be limited to underconditioned cows, while cows with optimal BCS should be maintained on an energy-restricted diet. Adipose tissue mRNA and microRNA expression further confirmed this hypothesis, and indicated a relationship between the immune and metabolic response of the adipose tissue underscoring the existence of a “self-regulatory” mechanism. The extensive analysis of the hepatic one-carbon metabolism and related pathways highlighted fundamental differences in the metabolic progression of grazing cows compared to their higher-yield counterpart in TMR-based systems. Results also indicated a greater flux through these pathways in optimally conditioned cows feed restricted prepartum. The third experiment examined the effect of over-feeding in both close-up and far-off periods on the adipose tissue transcriptome. Far-off over-feeding is usually a standard practice in seasonal grazing systems as, compared with TMR-fed cows, cows are thinner at the end of lactation. Adipose expression data revealed how overfed cows in the far-off period had greater adipogenesis, consistent with their rapid gain in BCS following dry-off, but a lower body fat mobilization in early lactation. The results indicated that neither strategy negatively affected the adaptations to lactation. However, to ensure a favorable transition, cows should be subjected to a small feed restriction in the close-up period, irrespective of far-off nutrition. Overall, results indicated a beneficial involvement of the immune system in the adaptation to lactation, and the possibility to regulate this process through prepartal BCS and nutrition management. As a result of the three studies, New Zealand farmers, through DairyNZ (the industry organization that represents all New Zealand dairy producers), are now discouraged to apply prophylactic pharmacological intervention early postpartum, in favour of nutritional management during the dry period. Our recommendation is for cows to be properly managed in late lactation and early dry period to attain optimal condition (e.g, BCS 5) by close-up (3 wks from calving). Subsequently cows will benefit from a controlled feed restriction (75–90% of requirements). On the other hand, cows in less than optimal condition (e.g. BCS ≤ 4) should be fed to requirements or slightly overfed (110-120% of requirements) before calving. This is an easily implementable strategy based on pasture allocation capable of benefitting the farmer with a minimum cost

Methyl donor supply to heat stress-challenged polymorphonuclear leukocytes from lactating Holstein cows enhances 1-carbon metabolism, immune response, and cytoprotective gene network abundance

[EN] Mechanisms controlling immune function of dairy cows are dysregulated during heat stress (HS). Methyl donor supply-methionine (Met) and choline (Chop-positively modulates innate immune function, particularly antioxidant systems of polymorphonuclear leukocytes (PMN). The objective of this study was to investigate the effect of Met and Chol supply in vitro on mRNA abundance of genes related to 1-carbon metabolism, inflammation, and immune function in short-term cultures of PMN isolated from mid-lactating Holstein cows in response to heat challenge. Blood PMN were isolated from 5 Holstein cows (153 +/- 5 d postpartum, 34.63 +/- 2.73 kg/d of milk production; mean +/- SD). The PMN were incubated for 2 h at thermal-neutral (37 degrees C; TN) or heat stress (42 degrees C; HS) temperatures with 3 levels of Chol (0, 400, or 800 mu g/mL) or 3 ratios of Lys:Met (Met; 3.6:1, 2.9:1, or 2.4:1). Supernatant concentrations of IL-1 beta, IL-6, and tumor necrosis factor-alpha were measured via bovine-specific ELISA. Fold-changes in mRNA abundance were calculated separately for Chol and Met treatments to obtain the fold-change response at 42 degrees C (HS) relative to 37 degrees C (TN). Data were subjected to ANOVA using PROC MIXED in SAS (SAS Institute Inc., Cary, NC). Orthogonal contrasts were used to determine the linear or quadratic effect of Met and Chol for mRNA fold-change and supernatant cytokine concentrations. Compared with PMN receiving 0 mu g of Chol/mL, heat-stressed PMN supplemented with Chol at 400 or 800 mu g/mL had greater fold-change in abundance of CBS, CSAD, GSS, GSR, and GPX1. Among genes associated with inflammation and immune function, fold-change in abundance of TLR2, TLR4, IRAK1, IL1B, and IL10 increased with 400 and 800 mu g of Chol/mL compared with PMN receiving 0 mu g of Chol/mL. Fold-change in abundance of SAHH decreased linearly at increasing levels of Met supply. A linear effect was detected for MPO, NFKB1, and SOD1 due to greater fold-change in abundance when Met was increased to reach Lys: Met ratios of 2.9:1 and 2.4:1. Although increasing Chol supply upregulated BAX, BCL2, and HSP70, increased Met supply only upregulated BAX. Under HS conditions, enhancing PMN supply of Chol to 400 mu g/mL effectively increased fold-change in abundance of genes involved in antioxidant production (conferring cellular processes protection from free radicals and reactive oxygen species), inflammatory signaling, and innate immunity. Although similar outcomes were obtained with Met supply at Lys:Met ratios of 2.9:1 and 2.4:1, the response was less pronounced. Both Chol and Met supply enhanced the cytoprotective characteristics of PMN through upregulation of heat shock proteins. Overall, the modulatory effects detected in the present experiment highlight an opportunity to use Met and particularly Chol supplementation during thermal stress.M. Vailati-Riboni was supported in part by Hatch funds under project ILLU-538-914, National Institute of Food and Agriculture (Washington, DC). The authors declare no conflict of interest.Lopreiato, V.; Vailati-Riboni, M.; Parys, C.; Fernández Martínez, CJ.; Minuti, A.; Loor, J. (2020). Methyl donor supply to heat stress-challenged polymorphonuclear leukocytes from lactating Holstein cows enhances 1-carbon metabolism, immune response, and cytoprotective gene network abundance. Journal of Dairy Science. 103(11):10477-10493. https://doi.org/10.3168/jds.2020-18638S104771049310311Abdelmegeid, M. K., Vailati-Riboni, M., Alharthi, A., Batistel, F., & Loor, J. J. (2017). Supplemental methionine, choline, or taurine alter in vitro gene network expression of polymorphonuclear leukocytes from neonatal Holstein calves. 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Heat Stress Elicits Different Responses in Peripheral Blood Mononuclear Cells from Brown Swiss and Holstein Cows. Journal of Dairy Science, 89(12), 4606-4612. doi:10.3168/jds.s0022-0302(06)72510-3Lecchi, C., Rota, N., Vitali, A., Ceciliani, F., & Lacetera, N. (2016). In vitro assessment of the effects of temperature on phagocytosis, reactive oxygen species production and apoptosis in bovine polymorphonuclear cells. Veterinary Immunology and Immunopathology, 182, 89-94. doi:10.1016/j.vetimm.2016.10.007Loos, H., Roos, D., Weening, R., & Houwerzijl, J. (1976). Familial deficiency of glutathione reductase in human blood cells. Blood, 48(1), 53-62. doi:10.1182/blood.v48.1.53.53Lopreiato, V., Vailati-Riboni, M., Bellingeri, A., Khan, I., Farina, G., Parys, C., & Loor, J. J. (2019). Inflammation and oxidative stress transcription profiles due to in vitro supply of methionine with or without choline in unstimulated blood polymorphonuclear leukocytes from lactating Holstein cows. 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Supplementation with rumen-protected methionine or choline during the transition period influences whole-blood immune response in periparturient dairy cows. Journal of Dairy Science, 100(5), 3958-3968. doi:10.3168/jds.2016-11812Yan, J., Meng, X., Wancket, L. M., Lintner, K., Nelin, L. D., Chen, B., … Liu, Y. (2012). Glutathione Reductase Facilitates Host Defense by Sustaining Phagocytic Oxidative Burst and Promoting the Development of Neutrophil Extracellular Traps. The Journal of Immunology, 188(5), 2316-2327. doi:10.4049/jimmunol.1102683Zhou, Z., Bulgari, O., Vailati-Riboni, M., Trevisi, E., Ballou, M. A., Cardoso, F. C., … Loor, J. J. (2016). Rumen-protected methionine compared with rumen-protected choline improves immunometabolic status in dairy cows during the peripartal period. Journal of Dairy Science, 99(11), 8956-8969. doi:10.3168/jds.2016-10986Zhou, Z., Ferdous, F., Montagner, P., Luchini, D. N., Corrêa, M. N., & Loor, J. J. (2018). 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Better postpartal performance in dairy cows supplemented with rumen-protected methionine compared with choline during the peripartal period

The onset of lactation in dairy cows is characterized by high output of methylated compounds in milk when sources of methyl group are in short supply. Methionine and choline (CHOL) are key methyl donors and their availability during this time may be limiting for milk production, hepatic lipid metabolism, and immune function. Supplementing rumen-protected Met and CHOL may improve overall performance and health of transition cows. The objective of this study was to evaluate the effect of supplemental rumen-protected Met and CHOL on performance and health of transition cows. Eighty-one multiparous Holstein cows were used in a randomized, complete, unbalanced block design with 2×2 factorial arrangement of Met (Smartamine M, Adisseo NA, Alpharetta, GA) and CHOL (ReaShure, Balchem Inc., New Hampton, NY) inclusion (with or without). Treatments (20 to 21 cows each) were control (CON), CON+Met (SMA), CON+CHOL (REA), and CON+Met+CHOL (MIX). From -50 to -21d before expected calving, all cows received the same diet (1.40Mcal of NEL/kg of DM) with no Met or CHOL. From -21d to calving, cows received the same close-up diet (1.52Mcal of NEL/kg of DM) and were assigned randomly to treatments (CON, SMA, REA, or MIX) supplied as top dresses. From calving to 30 DIM, cows were fed the same postpartal diet (1.71Mcal of NEL/kg of DM) and continued to receive the same treatments through 30 DIM. The Met supplementation was adjusted daily at 0.08% DM of diet and REA was supplemented at 60g/d. Incidence of clinical ketosis and retained placenta tended to be lower in Met-supplemented cows. Supplementation of Met (SMA, MIX) led to greater DMI compared with other treatments (CON, REA) in both close-up (14.3 vs. 13.2kg/d, SEM 0.3) and first 30d postpartum (19.2 vs. 17.2kg/d, SEM 0.6). Cows supplemented with Met (SMA, MIX) had greater yields of milk (44.2 vs. 40.4kg/d, SEM 1.2), ECM (44.6 vs. 40.5kg/d, SEM 1.0), and FCM (44.6 vs. 40.8kg/d, SEM 1.0) compared with other (CON, REA) treatments. Milk fat content did not differ in response to Met or CHOL. However, milk protein content was greater in Met-supplemented (3.32% vs. 3.14%, SEM 0.04%) but not CHOL-supplemented (3.27 vs. 3.19%, SEM 0.04%) cows. Supplemental CHOL led to greater blood glucose and insulin concentrations with lower glucose:insulin ratio. No Met or CHOL effects were detected for blood fatty acids or BHB, but a Met × time effect was observed for fatty acids due to higher concentrations on d 20. Results from the present study indicate that peripartal supplementation of rumen-protected Met but not CHOL has positive effects on cow performance

Inflammation and oxidative stress transcription profiles due to in vitro supply of methionine with or without choline in unstimulated blood polymorphonuclear leukocytes from lactating Holstein cows.

Neutrophils are the most important polymorphonuclear leukocytes (PMNL), representing the front-line defense involved in pathogen clearance upon invasion. As such, they play a pivotal role in immune and inflammatory responses. Isolated PMNL from 5 mid-lactating Holstein dairy cows were used to evaluate the in vitro effect of methionine (Met) and choline (Chol) supplementation on mRNA expression of genes related to the Met cycle and innate immunity. The target genes are associated with the Met cycle, cell signaling, inflammation, antimicrobial and killing mechanisms, and pathogen recognition. Treatments were allocated in a 3 × 3 factorial arrangement, including 3 Lys-to-Met ratios (L:M, 3.6:1, 2.9:1, or 2.4:1) and 3 levels of supplemental Chol (0, 400, or 800 μg/mL). Three replicates per treatment group were incubated for 2 h at 37°C and 5% atmospheric CO2. Both betaine-homocysteine S-methyltransferase and choline dehydrogenase were undetectable, indicating that PMNL (at least in vitro) cannot generate Met from Chol through the betaine pathway. The PMNL incubated without Chol experienced a specific state of inflammatory mediation [greater interleukin-1β (IL1B), myeloperoxidase (MPO), IL10, and IL6] and oxidative stress [greater cysteine sulfinic acid decarboxylase (CSAD), cystathionine gamma-lyase (CTH), glutathione reductase (GSR), and glutathione synthase (GSS)]. However, data from the interaction L:M × Chol indicated that this negative state could be overcome by supplementing additional Met. This was reflected in the upregulation of methionine synthase (MTR) and toll-like receptor 2 (TLR2); that is, pathogen detection ability. At the lowest level of supplemental Chol, Met downregulated GSS, GSR, IL1B, and IL6, suggesting it could reduce cellular inflammation and enhance antioxidant status. At 400 µg/mL Chol, supplemental Met upregulated PMNL recognition capacity [higher TLR4 and L-selectin (SELL)]. Overall, enhancing the supply of methyl donors to isolated unstimulated PMNL from mid-lactating dairy cows leads to a low level of PMNL activation and upregulates a cytoprotective mechanism against oxidative stress. Enhancing the supply of Met coupled with adequate Chol levels enhances the gene expression of PMNL pathogen-recognition mechanism. These data suggest that Chol supply to PMNL exposed to low levels of Met effectively downregulated the entire repertoire of innate inflammatory-responsive genes. Thus, Met availability in PMNL during an inflammatory challenge may be sufficient for mounting an appropriate biologic response

Cognitive-Behavioral Therapy: Current Paths in the Management of Obesity

The treatment of obesity and its related chronic symptoms is one of the major issues that world healthcare systems are facing today. Cognitive-behavioral therapy (CBT) is one of the most effective therapies in the treatment of dysfunctional eating behaviors. In the first part of the chapter, the phenomenon of obesity will be introduced; subsequently, the role of CBT into obesity treatment will be underlined. CBT’s core strategies will be presented and analyzed: goal setting, self-monitoring, stimulus control, problem solving technique, and cognitive restructuring technique. The use of these strategies and related results is a major issue, emphasizing the need for further studies on the phenomenon of obesity, given the excellent results available in the short term, with significant weight loss, but the difficulties in keeping the results achieved in the long run. Since obesity is a chronic condition, CBT treatments must focus on different outcomes, considering weight loss as a consequence of a change in the individual’s eating style rather than as a major and only result to be pursued. Finally, we will take into account the topic of motivation in the psychological treatment of obesity since patient’s motivation assessment seems to be a major prerequisite for successful weight loss therapy

Rumen-protected methionine compared with rumen-protected choline improves immunometabolic status in dairy cows during the peripartal period.

The immunometabolic status of peripartal cows is altered due to changes in liver function, inflammation, and oxidative stress. Nutritional management during this physiological state can affect the biological components of immunometabolism. The objectives of this study were to measure concentrations of biomarkers in plasma, liver tissue, and milk, and also polymorphonuclear leukocyte function to assess the immunometabolic status of cows supplemented with rumen-protected methionine (Met) or choline (CHOL). Forty-eight multiparous Holstein cows were used in a randomized complete block design with 2×2 factorial arrangement of Met (Smartamine M, Adisseo NA, Alpharetta, GA) and CHOL (ReaShure, Balchem Inc., New Hampton, NY) level (with or without). Treatments (12 cows each) were control (CON), no Met or CHOL; CON and Met (SMA); CON and CHOL (REA); and CON and Met and CHOL (MIX). From -50 to -21d before expected calving, all cows received the same diet [1.40Mcal of net energy for lactation (NEL)/kg of DM] with no Met or CHOL. From -21d to calving, cows received the same close-up diet (1.52Mcal of NEL/kg of DM) and were assigned randomly to each treatment. From calving to 30d, cows were on the same postpartal diet (1.71Mcal of NEL/kg of DM) and continued to receive the same treatments until 30d. The Met supplementation was adjusted daily at 0.08% DM of diet, and CHOL was supplemented at 60g/cow per day. Liver (-10, 7, 21, and 30d) and blood (-10, 4, 8, 20, and 30d) samples were harvested for biomarker analyses. Neutrophil and monocyte phagocytosis and oxidative burst were assessed at d 1, 4, 14, and 28d. The Met-supplemented cows tended to have greater plasma paraoxonase. Greater plasma albumin and IL-6 as well as a tendency for lower haptoglobin were detected in Met- but not CHOL-supplemented cows. Similarly, cows fed Met compared with CHOL had greater concentrations of total and reduced glutathione (a potent intracellular antioxidant) in liver tissue. Upon a pathogen challenge in vitro, blood polymorphonuclear leukocyte phagocytosis capacity and oxidative burst activity were greater in Met-supplemented cows. Overall, liver and blood biomarker analyses revealed favorable changes in liver function, inflammation status, and immune response in Met-supplemented cows

Photoprotection in intact cells of photosynthetic bacteria: quenching of bacteriochlorophyll fluorescence by carotenoid triplets.

Upon high light excitation in photosynthetic bacteria, various triplet states of pigments can accumulate leading to harmful effects. Here, the generation and lifetime of flash-induced carotenoid triplets (3Car) have been studied by observation of the quenching of bacteriochlorophyll (BChl) fluorescence in different strains of photosynthetic bacteria including Rvx. gelatinosus (anaerobic and semianaerobic), Rsp. rubrum, Thio. roseopersicina, Rba. sphaeroides 2.4.1 and carotenoid- and cytochrome-deficient mutants Rba. sphaeroides Ga, R-26, and cycA, respectively. The following results were obtained: (1) 3Car quenching is observed during and not exclusively after the photochemical rise of the fluorescence yield of BChl indicating that the charge separation in the reaction center (RC) and the carotenoid triplet formation are not consecutive but parallel processes. (2) The photoprotective function of 3Car is not limited to the RC only and can be described by a model in which the carotenoids are distributed in the lake of the BChl pigments. (3) The observed lifetime of 3Car in intact cells is the weighted average of the lifetimes of the carotenoids with various numbers of conjugated double bonds in the bacterial strain. (4) The lifetime of 3Car measured in the light is significantly shorter (1-2 mus) than that measured in the dark (2-10 mus). The difference reveals the importance of the dynamics of 3Car before relaxation. The results will be discussed not only in terms of energy levels of the 3Car but also in terms of the kinetics of transitions among different sublevels in the excited triplet state of the carotenoid

The effect of calving in the summer on the hepatic transcriptome of Holstein cows during the peripartal period

The liver is the main metabolic organ coordinating the adaptations that take place during the peripartal period of dairy cows. A successful transition into lactation, rather than management practices alone, depends on environmental factors such as temperature, season of parturition, and photoperiod. Therefore, we analyzed the effect of calving season on the hepatic transcriptome of dairy cows during the transition period. A total of 12 Holstein dairy cows were assigned into 2 groups based on calving season (6 cows March-April, spring; 6 cows June-July, summer, SU). The RNA was extracted from liver samples obtained at -30, 3, and 35 DIM via percutaneous biopsy and hybridized to the Agilent 44K Bovine (V2) Gene Expression Microarray (Agilent Technologies Inc., Santa Clara, CA). A quantitative PCR on 22 target genes was performed to verify and expand the analyses. A total of 4,307 differentially expressed genes were detected (false discovery rate ≤0.05) in SU compared with spring. Furthermore, 73 unique differentially expressed genes were detected in SU compared with spring cows after applying a fold-change threshold ≥3 and ≤-3. For Kyoto Encyclopedia of Genes and Genomes pathways analysis of differentially expressed genes, we used the dynamic impact approach. Ingenuity Pathway Analysis software was used to analyze upstream transcription regulators and perform gene network analysis. Among metabolic pathways, energy metabolism from lipids, carbohydrates, and amino acids was strongly affected by calving in SU, with a reduced level of fatty acid synthesis, oxidation, re-esterification, and synthesis of lipoproteins, leading to hepatic lipidosis. Glycan-synthesis was downregulated in SU cows probably as a mechanism to counteract the progression of this lipidosis. In contrast, calving in the SU resulted in upregulation of gluconeogenesis but also greater use of glucose as an energy source. Among nonmetabolic pathways, the heat-shock response was obviously activated in SU cows but was also associated with inflammatory and intracellular stress response. Furthermore, data support a recent finding that cows experience endoplasmic reticulum stress around parturition. Transcription regulator analysis revealed how metabolic changes are related to important regulatory mechanisms, including epigenetic modification. The holistic analyses of the liver transcriptome response to calving in the summer at high environmental temperatures underscore how transition cows should be carefully managed during this period, as they experience alterations in liver energy metabolism and inflammatory state increasing susceptibility to health disorders in early postpartum

The ACTyourCHANGE study protocol: promoting a healthy lifestyle in patients with obesity with Acceptance and Commitment Therapy-a randomized controlled trial

BackgroundAs treatment of choice in promoting psychological flexibility, Acceptance and Commitment Therapy (ACT) was found to be effective in several conditions, and among different populations, including weight management in individuals with obesity. However, the mechanism of action of psychological flexibility is less known. The aim of the present study is, within the context of a brief ACT intervention for behavioral change and behavioral maintenance of a healthy lifestyle in a sample of inpatients with obesity, to explore the effect of each subcomponent of the psychological flexibility model on treatment processes and outcomes.MethodsA randomized controlled trial will be conducted. Ninety Italian adult inpatients with obesity attending a rehabilitation program for weight loss will be randomly allocated into three experimental conditions targeting respectively each subcomponent of the psychological flexibility model: group Engage focused on values-oriented behaviors, group Openness focused on acceptance and cognitive defusion, and group Awareness focused on being present and aware of thoughts, feelings, and behaviors at every moment. Weight, BMI (kg/m(2)), the Psychological General Well-Being Inventory (PGWBI), the Outcome Questionnaire-45.2 (OQ-45.2), the Depression Anxiety and Stress Scale (DASS-21), the Difficulties in Emotion Regulation Scale (DERS), the Dutch Eating Behaviors Questionnaire (DEBQ), the Brief Values Inventory (BVI), the Committed Action Questionnaire (CAQ), the Italian-Cognitive Fusion Questionnaire (I-CFQ), the Five Facet Mindfulness Questionnaire (FFMQ), and the Acceptance and Action Questionnaire (AAQ-II) will be assessed at the beginning (time 0), at the end of psychological intervention (time 1), and after 3 (time 2) and 6months (time 3) and 9months (time 4) from discharge. During the following month after discharge, outpatients will be monitored in their adherence to a healthy lifestyle, using a wearable device.To assess the effectiveness of the intervention, mixed between-within 3 (conditions) x4 (times) repeated measure ANOVAs will be conducted to examine changes from time 0 to time 1, 2, 3, and 4 in means of weight, BMI, and means of scores PGWBI, OQ-45.2, DASS, DERS, DEBQ, AAQ-II, BVI, CAQ, I-CFQ, and FFMQ, between three groups.DiscussionThis study will contribute to clarify the mechanism of action of each subcomponent of the psychological flexibility model and understand its impact on the promotion of a healthy lifestyle.Trial registrationClinicalTrials.govNCT04474509. Registered on July 4, 202