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When cells in a primary tumor work together to invade into nearby tissue, this can lead to cell dissociations-cancer cells breaking off from the invading front-leading to metastasis. What controls the dissociation of cells, and whether they break off singly or in small groups? Can this be determined by cell-cell adhesion or chemotactic cues given to cells? We develop a physical model for this question, based on experiments that mimic aspects of cancer cell invasion using microfluidic devices with microchannels of different widths. Experimentally, most dissociation events ("ruptures") involve single cells breaking off, but we observe some ruptures of large groups ( ∼ 20 cells) in wider channels. The rupture probability is nearly independent of channel width. We recapitulate the experimental results with a phase field cell motility model by introducing three different cell states (follower, guided, and high-motility metabolically active leader cells) based on their spatial position. These leader cells may explain why single-cell rupture is the universal most probable outcome. Our simulation results show that cell-channel adhesion is necessary for cells in narrow channels to invade, and strong cell-cell adhesion leads to fewer but larger ruptures. Chemotaxis also influences the rupture behavior: Strong chemotaxis strength leads to larger and faster ruptures. Finally, we study the relationship between biological jamming transitions and cell dissociations. Our results suggest unjamming is necessary but not sufficient to create ruptures.

Cells tune adherens junction dynamics to regulate epithelial integrity in diverse (patho)physiological processes, including cancer metastasis. We hypothesized that the spatially confining architecture of peritumor stroma promotes metastatic cell dissemination by remodeling cell-cell adhesive interactions. By combining microfluidics with live-cell imaging, FLIM/FRET biosensors, and optogenetic tools, we show that confinement induces leader cell dissociation from cohesive ensembles. Cell dissociation is triggered by myosin IIA (MIIA) dismantling of E-cadherin cell-cell junctions, as recapitulated by a mathematical model. Elevated MIIA contractility is controlled by RhoA/ROCK activation, which requires distinct guanine nucleotide exchange factors (GEFs). Confinement activates RhoA via nucleocytoplasmic shuttling of the cytokinesis-regulatory proteins RacGAP1 and Ect2 and increased microtubule dynamics, which results in the release of active GEF-H1. Thus, confining microenvironments are sufficient to induce cell dissemination from primary tumors by remodeling E-cadherin cell junctions via the interplay of microtubules, nuclear trafficking, and RhoA/ROCK/MIIA pathway and not by down-regulating E-cadherin expression.

Pharmacogenomic testing can be used to guide medication selection in patients with major depressive disorder (MDD). Currently, there is no consensus on which gene or genes to consider in medication management. Here, we assessed the clinical validity of the combinatorial pharmacogenomic algorithm to predict sertraline blood levels in a subset of patients enrolled in the Genomics Used to Improve DEpression Decisions (GUIDED) trial. Patients who reported taking sertraline within ≤2 weeks of the screening blood draw were included. All patients received combinatorial pharmacogenomic testing, which included a weighted assessment of individual phenotypes for multiple pharmacokinetic genes relevant for sertraline (CYP2C19, CYP2B6, and CYP3A4). Sertraline blood levels were compared between phenotypes based on: 1) the pharmacokinetic portion of the combinatorial pharmacogenomic algorithm, and 2) individual genes. When evaluated separately, individual genes (for CYP2C19 and CYP2B6) and the combinatorial algorithm were significant predictors of sertraline blood levels. However, in multivariate analyses that included individual genes and the combinatorial pharmacogenomic algorithm, only the combinatorial pharmacogenomic algorithm remained a significant predictor of sertraline blood levels. These findings support the clinical validity of the combinatorial pharmacogenomic algorithm, in that it is a superior predictor of sertraline blood levels compared to individual genes., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Methylphenidate is among the most prescribed medications for treating attention-deficit/hyperactivity disorder (ADHD). However, nearly half of pediatric patients with ADHD do not respond to methylphenidate treatment. Pharmacogenetic testing can aid in identifying patients for whom methylphenidate is unlikely to be safe or effective, leading to improved methylphenidate outcomes and increased use of alternative treatment options for ADHD. This article aimed to summarize findings from studies of the ADRA2A gene variant, rs1800544, and its association with methylphenidate outcomes in ADHD., Methods: We systematically reviewed and meta-analyzed available literature on the impact of rs1800544 on methylphenidate outcomes in ADHD., Results: Fourteen studies met inclusion criteria for review, 9 of which were eligible for meta-analysis. The included studies compared methylphenidate outcomes in patients with ADHD categorized by rs1800544 genotype. G-allele carriers experienced significantly greater improvements in ADHD symptom scores (Swanson, Nolan, and Pelham Version-IV Scale or ADHD Rating Scale-IV) relative to noncarriers (odds ratio 3.08, 95% confidence interval 1.71-5.56, p  = .0002) and greater response rates as measured by a ≥50% improvement in symptom scores (odds ratio 2.68, 95% confidence interval 1.23-5.82, p  = .01); no significant difference in response rate as measured by Clinical Global Impressions score ≤2 was found. Stouffer's z -score method showed significant improvement across all methylphenidate outcomes in G-allele carriers relative to noncarriers ( z  = 3.03, p  = .002)., Conclusions: These findings suggest that carriers of rs1800544 may have improved ADHD outcomes following methylphenidate treatment. However, the extent to which these improvements are clinically impactful remain unclear. Additional studies are required to determine if rs1800544 carrier status should influence clinical recommendations for treatment of ADHD symptoms., (© 2021 The Authors.)

Pharmacogenomic tests used to guide clinical treatment for major depressive disorder (MDD) must be thoroughly validated. One important assessment of validity is the ability to predict medication blood levels, which reflect altered metabolism. Historically, the metabolic impact of individual genes has been evaluated; however, we now know that multiple genes are often involved in medication metabolism. Here, we evaluated the ability of individual pharmacokinetic genes (CYP2C19, CYP2D6, CYP3A4) and a combinatorial pharmacogenomic test (GeneSight Psychotropic®; weighted assessment of all three genes) to predict citalopram/escitalopram blood levels in patients with MDD. Patients from the Genomics Used to Improve DEpression Decisions (GUIDED) trial who were taking citalopram/escitalopram at screening and had available blood level data were included (N=191). In multivariate analysis of the individual genes and combinatorial pharmacogenomic test separately (adjusted for age, smoking status), the F statistic for the combinatorial pharmacogenomic test was 1.7 to 2.9-times higher than the individual genes, showing that it explained more variance in citalopram/escitalopram blood levels. In multivariate analysis of the individual genes and combinatorial pharmacogenomic test together, only the combinatorial pharmacogenomic test remained significant. Overall, this demonstrates that the combinatorial pharmacogenomic test was a superior predictor of citalopram/escitalopram blood levels compared to individual genes., Competing Interests: Declaration of Competing Interest Dr. Shelton has received research funding from Acadia Pharmaceuticals, Alkermes, Inc., Allergan, Assurex Health, Avanir Pharmaceuticals, Cerecor, Inc., Genomind, Intracellular Therapies, Janssen Pharmaceutica, Otsuka Pharmaceuticals, and Takeda Pharmaceuticals. Dr. Shelton has served as a consultant for Acadia Pharmaceuticals, Allergan Inc., Cerecor, Inc., Janssen Pharmaceutica, Lundbeck A/S, Takeda Pharmaceuticals. Dr. Parikh has received research funding from the Ontario Brain Institute, the Canadian Institutes of Health Research, the James and Ethel Flinn Foundation. Dr. Parikh has served as a consultant for Assurex Health. Dr. Parikh has received honoraria from Mensante Corporation, Takeda, and the Canadian Network for Mood and Anxiety Treatments (CANMAT). Dr. Parikh has equity in Mensante. Ms. Law is employed by Myriad Neuroscience (formerly Assurex Health). Dr. Rothschild has received research support from Allergan, AssureRx, Janssen, the National Institute of Mental Health, Takeda, Eli-Lilly, and Pfizer; Consultant: Alkermes, Eli Lilly and Company, GlaxoSmithKline, Myriad Genetics, Pfizer, SageTherapeutics, and Sanofi-Aventis. Dr. Rothschild receives royalties for the Rothschild Scale for Antidepressant Tachyphylaxis (RSAT)®; Clinical Manual for the Diagnosis and Treatment of Psychotic Depression, American Psychiatric Press, 2009; The Evidence-Based Guide to Antipsychotic Medications, American Psychiatric Press, 2010; The Evidence-Based Guide to Antidepressant Medications, American Psychiatric Press, 2012, and UpToDate®. Dr. Thase has received research support from Assurex Health, Acadia, Agency for Healthcare Research and Quality, Alkermes, Avanir, Forest, Intracellular, Janssen, National Institute of Mental Health, Otsuka, Patient-Centered Outcomes Research Institute, Takeda. Dr. Thase has served as a consultant for Acadia, Akilii, Alkermes, Allergan (Forest, Naurex), AstraZeneca, Cerecor, Eli Lilly, Fabre-Kramer, Gerson Lehrman Group, Guidepoint Global, Johnson & Johnson (Janssen, Ortho-McNeil), Lundbeck, MedAvante, Merck, Moksha8, Nestlé (PamLab), Novartis, Otsuka, Pfizer, Shire, Sunovion, Takeda. Dr. Thase receives royalties from American Psychiatric Press, Guilford Publications, Herald House, W.W. Norton & Company, Inc. Dr. Dunlop has received research support from Acadia, NIMH, Sage, Assurex Health, Axsome, Janssen, and Takeda. Dr. Dunlop has served has a consultant for Assurex Health and Aptinyx. Dr. DeBattista has received research support from Assurex Health and Brain Resources. Dr. Conway has received research support from LivaNova and Bristol-Myers Squibb, the Stanley Medical Research Institute, the National Institute of Mental Health, NeoSync Inc, The Taylor Family Institute for Innovative Psychiatric Research, The August Busch IV Foundation, and the Barnes-Jewish Hospital Foundation. Dr. Conway has received speaking fees from Bristol-Myers Squibb and Otsuka Pharmaceuticals. Dr. Conway has served as a research design consultant for LivaNova. Dr. Conway is a part time employee of the John Cochran Veterans Administration Hospital in St. Louis. Dr. Forester has received research funding from the National Institutes of Health, Rogers Family Foundation, Spier Family Foundation, Assurex Health, Eli Lilly, and Biogen. Dr. Forester has served as a consultant for Biogen. Dr. Macalusco has conducted clinical trials research as principal investigator for Acadia, Alkermes, Allergan, Assurex Health, Eisai, Lundbeck, Janssen, Naurex/Aptinyx, and Neurim; all study contracts and payments were made to Kansas University Medical Cancer Research Institute. Mr. Hain is employed by Myriad Neuroscience (formerly Assurex Health). Dr. Aguilar is employed by Myriad Neuroscience (formerly Assurex Health). Dr. Brown is employed by Myriad Genetics. Dr. Lewis is employed by Myriad Neuroscience (formerly Assurex Health). Dr. Jablonski is employed by Myriad Neuroscience (formerly Assurex Health). Dr. Greden has served as a scientific advisor for Janssen Pharmaceutical, Naurex (Allergan) Pharmaceutical, Cerecor Pharmaceutical, NeuralStem, Sage Therapeutics and Genomind. Dr. Greden has received reimbursement as a speaker for Assurex Health in 2014. All work done as an unpaid consultant to Assurex and Myriad., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

How migrating cells differentially adapt and respond to extracellular track geometries remains unknown. Using intravital imaging, we demonstrate that invading cells exhibit dorsoventral (top-to-bottom) polarity in vivo. To investigate the impact of dorsoventral polarity on cell locomotion through different confining geometries, we fabricated microchannels of fixed cross-sectional area, albeit with distinct aspect ratios. Vertical confinement, exerted along the dorsoventral polarity axis, induces myosin II-dependent nuclear stiffening, which results in RhoA hyperactivation at the cell poles and slow bleb-based migration. In lateral confinement, directed perpendicularly to the dorsoventral polarity axis, the absence of perinuclear myosin II fails to increase nuclear stiffness. Hence, cells maintain basal RhoA activity and display faster mesenchymal migration. In summary, by integrating microfabrication, imaging techniques, and intravital microscopy, we demonstrate that dorsoventral polarity, observed in vivo and in vitro, directs cell responses in confinement by spatially tuning RhoA activity, which controls bleb-based versus mesenchymal migration., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)

Cells migrate in vivo through complex confining microenvironments, which induce significant nuclear deformation that may lead to nuclear blebbing and nuclear envelope rupture. While actomyosin contractility has been implicated in regulating nuclear envelope integrity, the exact mechanism remains unknown. Here, we argue that confinement-induced activation of RhoA/myosin-II contractility, coupled with LINC complex-dependent nuclear anchoring at the cell posterior, locally increases cytoplasmic pressure and promotes passive influx of cytoplasmic constituents into the nucleus without altering nuclear efflux. Elevated nuclear influx is accompanied by nuclear volume expansion, blebbing, and rupture, ultimately resulting in reduced cell motility. Moreover, inhibition of nuclear efflux is sufficient to increase nuclear volume and blebbing on two-dimensional surfaces, and acts synergistically with RhoA/myosin-II contractility to further augment blebbing in confinement. Cumulatively, confinement regulates nuclear size, nuclear integrity, and cell motility by perturbing nuclear flux homeostasis via a RhoA-dependent pathway., (© 2019 Mistriotis et al.)

The sialoglycoprotein podocalyxin is absent in normal pancreas but is overexpressed in pancreatic cancer and is associated with poor clinical outcome. Here, we investigate the role of podocalyxin in migration and metastasis of pancreatic adenocarcinomas using SW1990 and Pa03c as cell models. Although ezrin is regarded as a cytoplasmic binding partner of podocalyxin that regulates actin polymerization via Rac1 or RhoA, we did not detect podocalyxin-ezrin association in pancreatic cancer cells. Moreover, depletion of podocalyxin did not alter actin dynamics or modulate Rac1 and RhoA activities in pancreatic cancer cells. Using mass spectrometry, bioinformatics analysis, coimmunoprecipitation, and pull-down assays, we discovered a novel, direct binding interaction between the cytoplasmic tail of podocalyxin and the large GTPase dynamin-2 at its GTPase, middle, and pleckstrin homology domains. This podocalyxin-dynamin-2 interaction regulated microtubule growth rate, which in turn modulated focal adhesion dynamics and ultimately promoted efficient pancreatic cancer cell migration via microtubule- and Src-dependent pathways. Depletion of podocalyxin in a hemispleen mouse model of pancreatic cancer diminished liver metastasis without altering primary tumor size. Collectively, these findings reveal a novel mechanism by which podocalyxin facilitates pancreatic cancer cell migration and metastasis. SIGNIFICANCE: These findings reveal that a novel interaction between podocalyxin and dynamin-2 promotes migration and metastasis of pancreatic cancer cells by regulating microtubule and focal adhesion dynamics., (©2019 American Association for Cancer Research.)

The adaptation of the skeleton to its mechanical environment is orchestrated by mechanosensitive osteocytes, largely by regulating the abundance of sclerostin, a secreted inhibitor of bone formation. We defined a microtubule-dependent mechanotransduction pathway that linked fluid shear stress to reactive oxygen species (ROS) and calcium (Ca 2+ ) signals that led to a reduction in sclerostin abundance in cultured osteocytes. We demonstrated that microtubules stabilized by detyrosination, a reversible posttranslational modification of polymerized α-tubulin, determined the stiffness of the cytoskeleton, which set the mechanoresponsive range of cultured osteocytes to fluid shear stress. We showed that fluid shear stress through the microtubule network activated NADPH oxidase 2 (NOX2)-generated ROS that target the Ca 2+ channel TRPV4 to elicit Ca 2+ influx. Furthermore, tuning the abundance of detyrosinated tubulin affected cytoskeletal stiffness to define the mechanoresponsive range of cultured osteocytes to fluid shear stress. Finally, we demonstrated that NOX2-ROS elicited Ca 2+ signals that activated the kinase CaMKII to decrease the abundance of sclerostin protein. Together, these discoveries may identify potentially druggable targets for regulating osteocyte mechanotransduction to affect bone quality., (Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

Background: Genetic and environmental factors contribute to the development of celiac disease (CD), but specific genetic predisposing factors remain poorly understood. One candidate is allele 2 of the hs1.2 enhancer within the immunoglobulin heavy chain region. In humans, there are four possible alleles and a previous study of an Italian cohort demonstrated a significantly increased frequency of allele 2 in patients with CD., Aims: The purpose of the current study was to determine if a similar association between allele 2 and CD exists in an American population from Dayton, OH., Methods: Subjects were screened for CD via esophagogastroduodenoscopy with duodenal biopsy. All biopsies were microscopically scored using a modified Marsh-Oberhuber classification. DNA was isolated from patients' buccal cells for hs1.2 genotype analysis using PCR., Results: Unlike the Italian cohort, allele 2 frequency was not significantly different in patients with histopathologic evidence of CD compared to patients without such evidence. However, our patient population as a whole demonstrated a significantly increased allele 2 frequency when compared to that previously reported within diverse ethnic populations., Conclusions: Since our comparative control patients do not necessarily reflect a healthy control population, an overall increase in allele 2 may reflect an association between allele 2 of the hs1.2 enhancer and a spectrum of gastrointestinal disorders., (Copyright © 2017 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.)

Here we introduce a Rapid Adaptable Portable In vitro Detection biosensor platform (RAPID) for detecting ligands that interact with nuclear hormone receptors (NHRs). The RAPID platform can be adapted for field use, allowing rapid evaluation of endocrine disrupting chemicals (EDCs) presence or absence in environmental samples, and can also be applied for drug screening. The biosensor is based on an engineered, allosterically activated fusion protein, which contains the ligand binding domain from a target NHR (human thyroid receptor β in this work). In vitro expression of this protein using cell-free protein synthesis (CFPS) technology in the presence of an EDC leads to activation of a reporter enzyme, reported through a straightforward colorimetric assay output. In this work, we demonstrate the potential of this biosensor platform to be used in a portable "just-add-sample" format for near real-time detection. We also demonstrate the robust nature of the cell-free protein synthesis component in the presence of a variety of environmental and human samples, including sewage, blood, and urine. The presented RAPID biosensor platform is significantly faster and less labor intensive than commonly available technologies, making it a promising tool for detecting environmental EDC contamination and screening potential NHR-targeted pharmaceuticals.

The objective of the current study was to determine the effects of concentrate feeding method on milk yield and composition, dry matter (DM) intake (DMI), body weight and body condition score, reproductive performance, energy balance, and blood metabolites of housed (i.e., accommodated indoors) dairy cows in early to mid lactation. Eighty-eight multiparous Holstein-Friesian cows were managed on 1 of 4 concentrate feeding methods (CFM; 22 cows per CFM) for the first 21 wk postpartum. Cows on all 4 CFM were offered grass silage plus maize silage (in a 70:30 ratio on a DM basis) ad libitum throughout the study. In addition, cows had a target concentrate allocation of 11 kg/cow per day (from d 13 postpartum) via 1 of 4 CFM, consisting of (1) offered on a flat-rate basis via an out-of-parlor feeding system, (2) offered based on individual cow's milk yields in early lactation via an out-of-parlor feeding system, (3) offered as part of a partial mixed ration (target intake of 5 kg/cow per day) with additional concentrate offered based on individual cow's milk yields in early lactation via an out-of-parlor feeding system, and (4) offered as part of a partial mixed ration containing a fixed quantity of concentrate for each cow in the group. In addition, all cows were offered 1 kg/cow per day of concentrate pellets via an in-parlor feeding system. We detected no effect of CFM on concentrate or total DMI, mean daily milk yield, concentrations and yields of milk fat and protein, or metabolizable energy intakes, requirements, or balances throughout the study. We also found no effects of CFM on mean or final body weight, mean or final body condition score, conception rates to first service, or any of the blood metabolites examined. The results of this study suggest that CFM has little effect on the overall performance of higher-yielding dairy cows in early to mid lactation when offered diets based on conserved forages., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Reproductive performance in the high-yielding dairy cow has severely decreased in the last 40 yr. The aim of this study was to compare the effectiveness of 4 nutritional strategies in improving the reproductive performance of high-yielding dairy cows. It was hypothesized that offering cows a high-starch ration in early lactation would enhance the onset of luteal activity, and that decreasing the severity of negative energy balance in the early postcalving period would improve reproductive parameters. Nutritional regimens aimed at improving fertility were applied to 96 Holstein-Friesian dairy animals. Upon calving, animals were allocated in a balanced manner to one of 4 dietary treatments. Primiparous animals were balanced according to live weight, body condition score and calving date. Multiparous animals were balanced according to parity, previous lactation milk yield, liveweight, body condition score and calving date. Treatment 1 was based on an industry best practice diet (control) to contain 170 g of crude protein/kg of dry matter. Treatment 2 was an individual cow feeding strategy, whereby the energy balance (EB) of individual animals was managed so as to achieve a predetermined target daily EB profile (±10 MJ/d). Treatment 3 was a high-starch/high-fat combination treatment, whereby an insulinogenic (high-starch) diet was offered in early lactation to encourage cyclicity and followed by a lipogenic (low-starch, high-fat) diet to promote embryo development. Treatment 4 was a low-protein diet, containing 140 g of crude protein/kg of dry matter, supplemented with protected methionine at an inclusion level of 40 g per animal per day. The nutritional strategies implemented in this study had no statistically significant effects on cow fertility measures, which included the onset of luteal activity, conception rate, in-calf rate, and the incidence of atypical cycles. The individual cow feeding strategy improved EB in early lactation but had no benefit on conception rate to first insemination. However, conception rate to second insemination, 100-d pregnancy rate (from the commencement of breeding), and overall pregnancy rate tended to be higher in this group. The high-starch/high-fat treatment tended to decrease the proportion of delayed ovulations and increase the proportion of animals cycling by d 50 postcalving. Animals that failed to conceive to first insemination had a significantly longer luteal phase in the first cycle postpartum and a longer inter-ovulatory interval in the second cycle postpartum. With regards to estrous behavior, results indicate that as the size of the sexually active group increased, the intensity of estrus and the expression of mounting or attempting to mount another cow also increased. Furthermore, cows that became pregnant displayed more intense estrous behavior than cows that failed to become pregnant., (Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The effects of the level of energy intake (high E and low E) offered before and after calving on body condition score at calving, production performance, and energy status in the first 250 d of lactation were evaluated in a 2 × 2 factorial design experiment involving 80 Holstein-Friesian dairy animals (40 primiparous and 40 multiparous). From d 80 until d 21 precalving, primiparous animals were offered either high or low pasture allowances. Thereafter, these animals were housed and had ad libitum access to a high energy density diet (high E) or restricted access [6 kg of dry matter (DM) per d] to a low energy density diet (low E), respectively, until calving. From d 100 until d 42 precalving, multiparous animals were offered either ad libitum or restricted (10 kg of DM/d) access to a late lactation diet, and thereafter, had ad libitum access to a high E diet or restricted access (7 kg of DM complete diet/d) to a low E diet, respectively, until calving. The forage to concentrate (F:C) ratios (DM basis) of these high E and low E diets [d 42 (d 21 in primiparous animals) until calving] were 64:36 and 83:17, respectively. Cows offered high E and low E precalving diets were allocated to either a high E or low E postcalving diet [F:C ratio (DM basis) of 30:70 and 70:30, respectively] and remained on these diets until d 250 of lactation. Multiparous animals offered a high E diet precalving had a significantly higher body condition score at calving than those offered the low E diet precalving. This effect was not evident in primiparous animals. Precalving diet had no significant effect on plasma nonesterified fatty acid concentrations during the last 3 wk precalving in primi- or multiparous animals. Primiparous animals offered a high E diet precalving had significantly higher postcalving plasma concentrations of nonesterified fatty acid, suggesting greater mobilization of body reserves. Primi- and multiparous animals offered a high E diet postcalving had a significantly higher dry matter intake, milk yield, and energy status postcalving compared with animals offered a low E diet postcalving. Milk yields of primiparous animals offered high E and low E diets postcalving were 29.7 and 24.8 kg/d, respectively, and milk yield of multiparous animals offered high E and low E diets postcalving were 33.5 and 28.2 kg/d, respectively. It is concluded that altering body condition score during the dry period is difficult but that specific dietary regimens applied precalving can have a significant influence on postcalving production and energy-related parameters., (Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Ninety autumn-calving Holstein dairy cows (45 primiparous and 45 multiparous; mean parity, 3.1) were allocated to 1 of 3 treatments; 173, 144, or 114 g of crude protein (CP)/kg of dry matter (DM) from calving until d 150 of lactation. On d 151 of lactation, half the animals receiving 114 g of CP/kg of DM went onto 144 g of CP/kg of DM, half of the animals receiving 144 g of CP/kg of DM went onto 173 g of CP/kg of DM, and half of the animals receiving 173 g of CP/kg of DM went onto 144 g of CP/kg of DM, with the remaining animals staying on their original treatments. This resulted in 6 treatments in mid to late lactation: 114/114; 144/144; 173/173; 114/144; 144/173; and 173/144 g of CP/kg of DM. Overall, 95.3% of cows intended for breeding conceived during a 6-mo breeding period. The average pregnancy rates to first service and first plus second service were 30.9% [standard error of the difference (SED), 0.05] and 56.7% (SED, 0.05) respectively. The average 100 d in-calf rate from the start of the breeding period was 70.5%, and at least one abnormal progesterone profile was observed in 62% of animals. An increase in dietary protein content decreased the requirement for treatment of metritis. There was no effect of dietary protein content on any of the reproductive or progesterone measures; for example, days to conception, calving interval, 100 d in-calf rate (from commencement of breeding), days to onset of luteal activity, average luteal phase, average interovulatory interval, or average interluteal interval. An increase in dietary protein content decreased the average daily energy balance. A more positive energy balance was associated with an increased requirement for the treatment of metritis in the current study. Cumulative energy balance was positively associated with conception. There was no effect of the concentration of plasma urea on any of the reproductive variables; however, the concentration of serum leptin was favorably associated with the time to progesterone increase above 3 ng/mL, which has been deemed essential for embryo survival. Additionally, the average peak concentration of progesterone and the duration of the average luteal phase were favorably associated with the interval from calving to conception. The latter relationships emphasize the importance of progesterone in achieving and maintaining pregnancy.

One of the main contributing factors to the decline in fertility in contemporary dairy farming is the inability to detect cows in estrus. In the current study, 90 Holstein dairy cows [45 primiparous and 45 multiparous (mean parity of 3.1)] were allocated to 1 of 3 treatments at calving; 173, 144, or 114 g of crude protein/kg of dry matter. Estrous behavior was recorded for one 30-min period every 12 h from calving until all animals reached 140 d postpartum. Behavioral activities were recorded according to a scoring system developed by Van Eerdenburg et al. (1996), with 9 key estrous behavioral activities each allocated a given number of points. If the total score allocated was greater than or equal to 50 points during a single or consecutive observational periods, then the animal was deemed to be in estrus. A total of 238 estrous cycles scored 50 points or above on the Van Eerdenburg et al. (1996) scale in this experiment, with 51.7% of these cycles being characterized as standing immobile on mounting. There were no direct effects of dietary protein content on estrous behavior; however, 3 significant stage of lactation x protein treatment interactions occurred for the behavioral activities (mucous discharge, chin resting, and mounting the head side of another cow), but no consistent trends were apparent from the predicted means. There was a significant influence of parity on the frequency of mounting the head side of another cow and total number of behavior activities displayed per estrous cycle. In both cases multiparous animals displayed fewer behavioral activities than primiparous animals. An increase in the size of the sexually active group (animals in estrus at the same time, up to 5) significantly increased the expression of mounting or attempting to mount another cow, the number of cycles in which standing immobile on being mounted was observed, the total estrous score and the proportion of cyclic animals that were diagnosed as being in estrus. The most frequent behavioral activity displayed was chin resting (89.5% of cycles), and the most reliable behavior was standing immobile on mounting (when expressed 96.4% of cows were in estrus). The most dependable (function of reliability and frequency displayed) sign of estrus was mounting or attempting to mount another cow. This behavior was expressed in 83% of cycles, and when expressed, 89% of animals were in estrus.

Ninety autumn-calving Holstein dairy cows [45 primiparous and 45 multiparous (mean parity, 3.1)] were allocated to 1 of 3 dietary crude protein (CP) concentrations: 173, 144, or 114 g of CP/kg of DM, from calving until d 150 of lactation. On d 151, half of the animals in each treatment were allocated an alternative dietary protein concentration. Half of the animals receiving 114 g of CP/kg of DM went onto 144 g of CP/kg of DM; half of the animals receiving 144 g of CP/kg of DM went onto 173 g of CP/kg of DM; and half of the animals receiving 173 g of CP/kg of DM went onto 144 g of CP/kg of DM, with the remaining animals staying on their original treatment. This resulted in 6 treatments in the mid to late lactation period: 114/114, 144/144, 173/173, 114/144, 144/173, and 173/144 g of CP/kg of DM. An increase in dietary CP concentration significantly increased milk, fat, and protein yield in early lactation (d 1 to 150). Dry matter intake was also increased with increased dietary protein concentration; however, this was not significant between 144 and 173 g of CP/kg of DM. Increased dietary CP significantly increased plasma urea, albumin, and total protein concentrations but had no significant effect on NEFA, leptin, or IGF-1 concentrations. Decreasing the dietary CP concentration in mid-late lactation (d 151 to 305) from 173 to 144 g/kg of DM had no significant effect on milk yield, dry matter intake, or milk fat and protein yield, compared with animals that remained on 173 g of CP/kg of DM throughout lactation. Increasing dietary CP concentration from 144 to 173 g/kg of DM significantly increased dry matter intake compared with animals that remained on the 144 g of CP/kg of DM throughout lactation. There were no significant dietary treatment effects on live weight or body condition score change throughout the experiment. Results of this study indicate that high protein diets (up to 173 g of CP/kg of DM) improved feed intake and animal performance in early lactation (up to d 150), but thereafter, protein concentration can be reduced to 144 g of CP/kg of DM with no detrimental effects on animal performance.