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Sample records for protein pthrp lactation

  1. Mass Spectrometric Immunoassay for Parathyroid Hormone Related Protein (PTHrP)

    SciTech Connect

    Zheng, K.; Rivera, J.D.; Vogel, J.S.; Buchholz, B.A.; Burton, D.W.; Deftos, L.J.; Herold, D.A.; Fitzgerald, R.L.

    2000-06-16

    Many cancers, including prostate, breast and lung express parathyroid hormone related protein (PTHrP). Despite the common tumor overexpression of PTHrP, serum levels of PTHrP are not commonly elevated in affected patients. They postulate that the reasons for the discrepancy between tissue and serum measurements of PTHrP are the inadequate sensitivity and specificity of current PTHrP serum assays. To improve the clinical value of PTHrP serum assays for the cancer patient, they are developing a new generation of novel and ultrasensitive PTHrP serum immunoassays based on immunoaffinity purification, nanospray liquid chromatography tandem mass spectrometry (LC/MS/MS) and accelerator mass spectrometry (AMS).

  2. Parathyroid hormone-related protein (pthrp) is a gravisensor for lung and bone.

    NASA Astrophysics Data System (ADS)

    Torday, J.

    Parathyroid Hormone-related Protein (PTHrP) and its receptor represent a stretch- sensitive paracrine signaling mechanism (Torday, 1999) that may sense gravity. PTHrP has been shown to be essential for the development and homeostatic regulation of lung (Rubin et al, 2000) and bone (Kronenberg et al, 1994). Since both lung and bone structure and function are affected by microgravity, we hypothesized that microgravity down-regulates PTHrP signaling. To test this hypothesis, we suspended lung and bone cells in the microgravity environment of a rotating wall vessel apparatus, which simulates microgravity, for up to 72 hours. During the first 6-8 hours, PTHrP expression fell precipitously, decreasing by 80-90%; during the subsequent 64-66 hours, PTHrP expression remained at this newly established level. PTHrP production decreased from 5 pmol/ml/3hours to undetectable levels in culture medium from microgravity-exposed cells. The cells were then put back in culture at unit gravity for 24hours, and PTHrP expression and production returned to normal levels. Based on these findings, we have obtained bones from rats flown in space for 2 weeks (mission SLS-2, provided courtesy of the Biospecimen Facility, Ames Research Center, NASA, as a result of a peer-reviewed proposal). Analysis of PTHrP expression by femurs and tibias from these animals (n=5) revealed that PTHrP expression was 60% lower than in bones from ground-based rats. Interestingly, there were no differences in PTHrP exp ression by parietal bones, indicating that the effect of weightlessness on PTHrP expression is due to the unweighting of weight-bearing bones. This finding is consistent with other studies of microgravity-induced osteoporosis. The loss of the PTHrP signaling mechanism may be corrected using chemical agents that up-regulate this pathway.

  3. Identification, molecular characterization, and tissue expression of parathyroid hormone-related protein gene (PTHrP) from water buffalo (Bubalus bubalis).

    PubMed

    Liu, J; Qian, L D; Huo, J L; Bi, B L; Li, D L; Wang, S F; Chen, T; Li, L J; Mao, H M; Miao, Y W

    2015-03-27

    Parathyroid hormone-related protein (PTHrP) is involved in the deposition of milk calcium in mammal lactation, but its role in buffalo is unclear. In this study, the full-length coding sequence of the water buffalo PTHrP gene was first isolated using reverse transcription-polymerase chain reaction. The protein was then subjected to molecular characterization using bioinformatic methods, and the tissue expression pattern was further assayed by semi-quantitative reverse-transcription polymerase chain reaction. The water buffalo PTHrP gene contains an open reading frame of 534 base pairs encoding a polypeptide of 177 amino acid residues, a theoretical molecular weight of 20.32 kDa, and an isoelectric point of 10.00. In addition, water buffalo PTHrP was predicted to contain a signal peptide, a typical hydrophobic region with no hydrophobic transmembrane regions, and to exert its function in the cell nucleus. A conserved domain of parathyroid superfamily from amino acids 34-114 was observed in the polypeptide. Sequence comparison and the phylogenetic analysis showed that the sequence of the water buffalo PTHrP protein shared high homology with that of other mammals, particularly cattle and goat. Among the 16 tissues examined, the PTHrP gene was only expressed in adipose tissue, placenta, uterine wall, hypophysis, and mammary gland tissue, but gene expression levels were higher in the uterus wall and adipose tissue. The results of this study suggest that the PTHrP gene plays an important role in the deposition of milk calcium of water buffalo.

  4. Structural requirements for the action of parathyroid hormone-related protein (PTHrP) on bone resorption by isolated osteoclasts

    SciTech Connect

    Evely, R.S.; Bonomo, A.; Schneider, H.G.; Moseley, J.M.; Gallagher, J.; Martin, T.J. )

    1991-01-01

    Parathyroid hormone-related protein (PTHrP) plays a major role in the syndrome of humoral hypercalcemia of malignancy (HHM) by its actions on bone and kidney. In this study an isolated osteoclast bone resorption assay was used to investigate the actions of this peptide and the structure-activity relationships for its resorption effect. As with PTH, neither synthetic nor recombinant PTHrP preparations stimulated resorption within highly purified osteoclast populations. Resorption was stimulated only in the presence of contaminating osteoblasts or in cocultures with the osteoblast-like cell line UMR-106. In the presence of osteoblasts PTHrP-(1-34) and PTHrP-(1-84) stimulated bone resorption in a dose-dependent manner with a potency comparable to that of PTH-(1-34) on a molar basis. The biologic activity of the PTHrP was shown to reside in the first 34 amino acids, and within that region the structural requirements for promotion of osteoclastic resorption resembled closely those for promotion of cyclic AMP formation in osteoblast-like cells. Using emulsion autoradiography with iodinated PTHrP-(1-34) and PTHrP-(1-84) on mixed bone cell preparations from neonatal rats, specific binding was demonstrated only to osteoblasts, not to osteoclasts. These results clearly demonstrate that PTHrP is a potent stimulator of bone resorption and that these effects are, like those of PTH, mediated by initial actions upon cells of the osteoblast lineage.

  5. The Role of Parathyroid Hormone-Related Protein (PTHrP) in Osteoblast Response to Microgravity: Mechanistic Implications for Osteoporosis Development

    PubMed Central

    Camirand, Anne; Goltzman, David; Gupta, Ajay; Kaouass, Mohammadi; Panda, Dibyendu; Karaplis, Andrew

    2016-01-01

    Prolonged skeletal unloading through bedrest results in bone loss similar to that observed in elderly osteoporotic patients, but with an accelerated timeframe. This rapid effect on weight-bearing bones is also observed in astronauts who can lose up to 2% of their bone mass per month spent in Space. Despite the important implications for Spaceflight travelers and bedridden patients, the exact mechanisms involved in disuse osteoporosis have not been elucidated. Parathyroid hormone-related protein (PTHrP) regulates many physiological processes including skeletal development, and has been proposed as a mechanosensor. To investigate the role of PTHrP in microgravity-induced bone loss, trabecular and calvarial osteoblasts (TOs and COs) from Pthrp +/+ and -/- mice were subjected to actual Spaceflight for 6 days (Foton M3 satellite). Pthrp +/+, +/- and -/- osteoblasts were also exposed to simulated microgravity for periods varying from 6 days to 6 weeks. While COs displayed little change in viability in 0g, viability of all TOs rapidly decreased in inverse proportion to PTHrP expression levels. Furthermore, Pthrp+/+ TOs displayed a sharp viability decline after 2 weeks at 0g. Microarray analysis of Pthrp+/+ TOs after 6 days in simulated 0g revealed expression changes in genes encoding prolactins, apoptosis/survival molecules, bone metabolism and extra-cellular matrix composition proteins, chemokines, insulin-like growth factor family members and Wnt-related signalling molecules. 88% of 0g-induced expression changes in Pthrp+/+ cells overlapped those caused by Pthrp ablation in normal gravity, and pulsatile treatment with PTHrP1-36 not only reversed a large proportion of 0g-induced effects in Pthrp+/+ TOs but maintained viability over 6-week exposure to microgravity. Our results confirm PTHrP efficacy as an anabolic agent to prevent microgravity-induced cell death in TOs. PMID:27463808

  6. The Role of Parathyroid Hormone-Related Protein (PTHrP) in Osteoblast Response to Microgravity: Mechanistic Implications for Osteoporosis Development.

    PubMed

    Camirand, Anne; Goltzman, David; Gupta, Ajay; Kaouass, Mohammadi; Panda, Dibyendu; Karaplis, Andrew

    2016-01-01

    Prolonged skeletal unloading through bedrest results in bone loss similar to that observed in elderly osteoporotic patients, but with an accelerated timeframe. This rapid effect on weight-bearing bones is also observed in astronauts who can lose up to 2% of their bone mass per month spent in Space. Despite the important implications for Spaceflight travelers and bedridden patients, the exact mechanisms involved in disuse osteoporosis have not been elucidated. Parathyroid hormone-related protein (PTHrP) regulates many physiological processes including skeletal development, and has been proposed as a mechanosensor. To investigate the role of PTHrP in microgravity-induced bone loss, trabecular and calvarial osteoblasts (TOs and COs) from Pthrp +/+ and -/- mice were subjected to actual Spaceflight for 6 days (Foton M3 satellite). Pthrp +/+, +/- and -/- osteoblasts were also exposed to simulated microgravity for periods varying from 6 days to 6 weeks. While COs displayed little change in viability in 0g, viability of all TOs rapidly decreased in inverse proportion to PTHrP expression levels. Furthermore, Pthrp+/+ TOs displayed a sharp viability decline after 2 weeks at 0g. Microarray analysis of Pthrp+/+ TOs after 6 days in simulated 0g revealed expression changes in genes encoding prolactins, apoptosis/survival molecules, bone metabolism and extra-cellular matrix composition proteins, chemokines, insulin-like growth factor family members and Wnt-related signalling molecules. 88% of 0g-induced expression changes in Pthrp+/+ cells overlapped those caused by Pthrp ablation in normal gravity, and pulsatile treatment with PTHrP1-36 not only reversed a large proportion of 0g-induced effects in Pthrp+/+ TOs but maintained viability over 6-week exposure to microgravity. Our results confirm PTHrP efficacy as an anabolic agent to prevent microgravity-induced cell death in TOs.

  7. Current perspectives on parathyroid hormone (PTH) and PTH-related protein (PTHrP) as bone anabolic therapies.

    PubMed

    Esbrit, Pedro; Alcaraz, María José

    2013-05-15

    Osteoporosis is characterized by low bone mineral density and/or poor bone microarchitecture leading to an increased risk of fractures. The skeletal alterations in osteoporosis are a consequence of a relative deficit of bone formation compared to bone resorption. Osteoporosis therapies have mostly relied on antiresorptive drugs. An alternative therapeutic approach for osteoporosis is currently available, based on the intermittent administration of parathyroid hormone (PTH). Bone anabolism caused by PTH therapy is mainly accounted for by the ability of PTH to increase osteoblastogenesis and osteoblast survival. PTH and PTH-related protein (PTHrP)-an abundant local factor in bone- interact with the common PTH type 1 receptor with similar affinities in osteoblasts. Studies mainly in osteoporosis rodent models and limited data in postmenopausal women suggest that N-terminal PTHrP peptides might be considered a promising bone anabolic therapy. In addition, putative osteogenic actions of PTHrP might be ascribed not only to its N-terminal domain but also to its PTH-unrelated C-terminal region. In this review, we discuss the underlying cellular and molecular mechanisms of the anabolic actions of PTH and the similar potential of PTH-related protein (PTHrP) to increase bone mass and improve bone regeneration. Copyright © 2013 Elsevier Inc. All rights reserved.

  8. Paraneoplastic hormones: parathyroid hormone-related protein (PTHrP) and erythropoietin (EPO) are related to vascular endothelial growth factor (VEGF) expression in clear cell renal cell carcinoma.

    PubMed

    Feng, Chen-chen; Ding, Guan-xiong; Song, Ning-hong; Li, Xuan; Wu, Zhong; Jiang, Hao-wen; Ding, Qiang

    2013-12-01

    To investigate the correlation between parathyroid hormone-related protein (PTHrP), erythropoietin (EPO), and vascular endothelial growth factor (VEGF) expression in clear cell renal cell carcinoma (ccRCC). Immunohistochemical studies on PTHrP, EPO and VEGF were performed in 249 patients with ccRCC. Serum calcium level and haematocrit were analyzed. The expression of the factors and clinicopathological parameters were studied statistically for possible correlations. The incidence for hypercalcaemia and polycythaemia were 15.3% and 2.0% respectively. Expression of PTHrP, EPO, and VEGF were respectively related to advanced stage (P < 0.0001 respectively). PTHrP was not related to tumour grade. Expressions of EPO and VEGF were correlated to tumour grade significantly. All factors were expressed higher in hypercalcaemic patients. PTHrP, EPO, and VEGF were positively correlated with each other in non-hypercalcaemic patients yet not in hypercalcaemic ones. PTHrP and EPO are related to VEGF expression and to the progression of ccRCC. This finding offers us new insight on the behaviour of ccRCC and offers possible targets in RCC treatment.

  9. PTHrP and skeletal development.

    PubMed

    Kronenberg, Henry M

    2006-04-01

    Parathyroid hormone-related protein (PTHrP) participates in the regulation of endochondral bone development. After the cartilage mold is established in fetal life, perichondrial cells and chondrocytes at the ends of the mold synthesize PTHrP. This ligand then acts on PTH/PTHrP receptors on chondrocytes. As chondrocytes go through a program of proliferation and then further differentiation into post-mitotic, hypertrophic chondrocytes, PTHrP action keeps chondrocytes proliferating and delays their further differentiation. Indian hedgehog (Ihh) is synthesized by chondrocytes that have just stopped proliferating and is required for synthesis of PTHrP. The feedback loop between PTHrP and Ihh serves to regulate the pace of chondrocyte differentiation and the sites at which perichondrial cells first differentiate into osteoblasts. Activation of the PTH/PTHrP receptor leads to stimulation of both Gs and Gq family heterotrimeric G proteins. Genetic analyses demonstrate that Gs activation mediates the action of PTHrP to keep chondrocytes proliferating, while Gq activation opposes this action. Downstream from Gs activation, synthesis of the cyclin-cdk inhibitor, p57, is suppressed, thereby increasing the pool of proliferating chondrocytes. PTHrP's actions to delay chondrocyte differentiation are mediated by the phosphorylation of the transcription factor, SOX9, and by suppression of synthesis of mRNA encoding the transcription factor, Runx2. These pathways and undoubtedly others cooperate to regulate the pace of differentiation of growth plate chondrocytes in response to PTHrP.

  10. Acoustic wave biosensor for the detection of the breast and prostate cancer metastasis biomarker protein PTHrP.

    PubMed

    Crivianu-Gaita, Victor; Aamer, Mohamed; Posaratnanathan, Roy T; Romaschin, Alexander; Thompson, Michael

    2016-04-15

    There are currently no biosensors that are able to reliably detect the process of cancer metastasis. We describe the first label-free real-time ultra-high frequency acoustic wave biosensor prototype capable of detecting the breast and prostate cancer metastasis biomarker, parathyroid hormone-related peptide (PTHrP). Two different linkers - 11-trichlorosilyl-undecanoic acid pentafluorophenyl ester (PFP) and S-(11-trichlorosilyl-undecanyl)-benzothiosulfonate (TUBTS) - were used to immobilize whole anti-PTHrP antibodies and Fab' fragments to surfaces as biorecognition elements. The biosensor surfaces were optimized using X-ray photoelectron spectroscopy (XPS) and the ultra-high frequency electromagnetic piezoelectric acoustic sensor (EMPAS). One optimized whole antibody-based surface (PFP/protein G'/whole antibodies/ethanolamine) and one optimized Fab' fragment-based surface (TUBTS/Fab' fragments) were tested as biosensors. It was determined that an in-line injection of bovine serum albumin prior to analyte injection yielded the most minimally fouling surfaces. Each surface was tested with no mass amplification and with sandwich-type secondary antibody mass amplification. The whole antibody-based mass-amplified biosensor yielded the lowest limit of detection (61 ng/mL), highest sensitivity, and a linear range from 61 ng/mL to 100 μg/mL. However, the Fab' fragment-based biosensor displayed better regenerability as a loss of ~20% of the initial analyte signal intensity was observed with each subsequent injection. The whole antibody-based biosensor was only capable of producing an analyte signal in the first injection.

  11. Parathyroid hormone-related protein (PTHrP) is responsible for production of bone metastasis, but not visceral metastasis, by human small cell lung cancer SBC-5 cells in natural killer cell-depleted SCID mice.

    PubMed

    Miki, Toyokazu; Yano, Seiji; Hanibuchi, Masaki; Kanematsu, Takanori; Muguruma, Hiroaki; Sone, Saburo

    2004-02-10

    We previously established an osteolytic bone metastasis model with multiorgan dissemination in natural killer (NK) cell-depleted severe combined immunodeficient (SCID) mice using human small cell lung cancer cells (SBC-5), which highly express the parathyroid hormone-related protein (PTHrP). In our present study, we evaluated the role of PTHrP on bone metastasis by SBC-5 cells using anti-PTHrP neutralizing antibody (Ab). Anti-PTHrP Ab did not affect the proliferation or cytokine production of SBC-5 cells in vitro. Repeated intravenous injection with anti-PTHrP Ab inhibited the formation of bone metastasis in a dose-dependent manner, while the same treatment had no significant effect on the metastasis to visceral organs (lung, liver, kidney and lymph node). In addition, treatment with anti-PTHrP Ab improved the elevated serum calcium level, associated with inhibition of osteolytic bone metastasis, suggesting that anti-PTHrP Ab inhibited bone metastasis via suppression of bone resorption probably by neutralizing PTHrP. These findings suggest that PTHrP is essential for bone metastasis, but not visceral metastasis, by small cell lung cancer SBC-5 cells.

  12. Modification of the analysis of parathyroid hormone-related protein in milk and concentrations of this protein in commercial milk and milk products in Japan.

    PubMed

    Onda, K; Yamaguchi, M; Ohashi, M; Sato, R; Ochiai, H; Iriki, T; Wada, Y

    2010-05-01

    Parathyroid hormone-related protein (PTHrP), which causes hypercalcemia associated with malignant tumors, is known to be present in milk. Gene expression of PTHrP in the mammary gland increases markedly during parturition and with the onset of lactation. Even when circulating PTHrP levels are extremely low or below the detection limit, milk PTHrP levels are remarkably high. Parathyroid hormone-related protein derived from the mammary gland is assumed to play a role in maintaining the maternal calcium homeostasis and calcium transport from blood to milk. In previous studies that determined the PTHrP concentrations in milk, the pretreatments and diluent composition were not standardized. Here, we investigated the effect of various pretreatment procedures and diluent constitutions and the consequent PTHrP concentrations in commercial milk and milk products in Japan. Significant differences were found in PTHrP concentrations in raw milk samples subjected to different combinations of pretreatments (mixing, centrifugation, acidification, and heating) and diluents (0pM standard solution of PTHrP, plasma treated with protease inhibitors, and original diluent). We measured the PTHrP concentrations in normal liquid milk, processed milk, milk drinks, formulated milk powders, and skim milk powder by using the appropriate combination of pretreatment (acidification) and diluent (plasma treated with protease inhibitors). The PTHrP concentration in normal liquid milk, processed milk, and skim milk powder was as high as that in raw milk (>5nM), whereas that in milk drinks differed considerably. The PTHrP concentration in infant formulas (<2nM) was lower than that in the other milk products. These results indicate that a certain amount of PTHrP is ingested when milk and milk products are consumed.

  13. Osteoblast-Derived PTHRP and Breast Cancer Bone Metastasis

    DTIC Science & Technology

    2004-11-01

    Protein Structure 12.1.1 Background ’This sLudy was .supported by grants from the Susan G. Komen Foundation (KG 1 00766) and the Canadian Instimtes for...Parathyroid Hormone Related Protein (PTHrP) in Tumor Progression 147 lA 1B lC 2 3 4 5 6 Fig. 12.1 Structure of human parathyroid hormone-related protein ...Gene Sequence and Protein Structure The Pthrp gene is localized to the short arm of chromosome 12 in humans [26] whereas the PTH gene is on the

  14. PTHrP promotes malignancy of human oral cancer cell downstream of the EGFR signaling

    SciTech Connect

    Yamada, Tamaki; Tsuda, Masumi; Ohba, Yusuke Kawaguchi, Hideaki; Totsuka, Yasunori; Shindoh, Masanobu

    2008-04-11

    Parathyroid hormone-related protein (PTHrP) is detected in many aggressive tumors and involved in malignant conversion; however, the underlying mechanism remains obscure. Here, we identified PTHrP as a mediator of epidermal growth factor receptor (EGFR) signaling to promote the malignancies of oral cancers. PTHrP mRNA was abundantly expressed in most of the quiescent oral cancer cells, and was significantly upregulated by EGF stimulation via ERK and p38 MAPK. PTHrP silencing by RNA interference, as well as EGFR inhibitor AG1478 treatment, significantly suppressed cell proliferation, migration, and invasiveness. Furthermore, combined treatment of AG1478 and PTHrP knockdown achieved synergistic inhibition of malignant phenotypes. Recombinant PTHrP substantially promoted cell motility, and rescued the inhibition by PTHrP knockdown, suggesting the paracrine/autocrine function of PTHrP. These data indicate that PTHrP contributes to the malignancy of oral cancers downstream of EGFR signaling, and may thus provide a therapeutic target for oral cancer.

  15. A 7-day continuous infusion of PTH or PTHrP suppresses bone formation and uncouples bone turnover.

    PubMed

    Horwitz, Mara J; Tedesco, Mary Beth; Sereika, Susan M; Prebehala, Linda; Gundberg, Caren M; Hollis, Bruce W; Bisello, Alessandro; Garcia-Ocaña, Adolfo; Carneiro, Raquel M; Stewart, Andrew F

    2011-09-01

    Human in vivo models of primary hyperparathyroidism (HPT), humoral hypercalcemia of malignancy (HHM), or lactational bone mobilization for more than 48 hours have not been described previously. We therefore developed 7-day continuous-infusion models using human parathyroid hormone(1-34) [hPTH(1-34)] and human parathyroid hormone-related protein(1-36) [hPTHrP(1-36)] in healthy human adult volunteers. Study subjects developed sustained mild increases in serum calcium (10.0 mg/dL), with marked suppression of endogenous PTH(1-84). The maximal tolerated infused doses over a 7-day period (2 and 4 pmol/kg/h for PTH and PTHrP, respectively) were far lower than in prior, briefer human studies (8 to 28 pmol/kg/h). In contrast to prior reports using higher PTH and PTHrP doses, both 1,25-dihydroxyvitamin D(3) [1,25(OH)(2) D(3) ] and tubular maximum for phosphorus (TmP/GFR) remained unaltered with these low doses despite achievement of hypercalcemia and hypercalciuria. As expected, bone resorption increased rapidly and reversed promptly with cessation of the infusion. However, in contrast to events in primary HPT, bone formation was suppressed by 30% to 40% for the 7 days of the infusions. With cessation of PTH and PTHrP infusion, bone-formation markers abruptly rebounded upward, confirming that bone formation is suppressed by continuous PTH or PTHrP infusion. These studies demonstrate that continuous exposure of the human skeleton to PTH or PTHrP in vivo recruits and activates the bone-resorption program but causes sustained arrest in the osteoblast maturation program. These events would most closely mimic and model events in HHM. Although not a perfect model for lactation, the increase in resorption and the rebound increase in formation with cessation of the infusions are reminiscent of the maternal skeletal calcium mobilization and reversal that occur following lactation. The findings also highlight similarities and differences between the model and HPT. Copyright

  16. Nanomechanical properties of globular proteins: lactate oxidase.

    PubMed

    Parra, Ana; Casero, Elena; Lorenzo, Encarnación; Pariente, Félix; Vázquez, Luis

    2007-02-27

    We report on the study of the nanomechanical properties of a lactate oxidase (LOx) monolayer immobilized on gold substrates by atomic force microscopy techniques operating under buffer conditions. Topographical contact mode imaging evidenced the protein deformation under the applied tip load. We performed approaching force curves with both stiff and soft cantilevers by imposing maximum loads of 1.6 nN and 400 pN, respectively. We found that the experimental data were well fitted by the Hertz model for a conical indenter. The use of two types of cantilevers allowed us to check further the consistency of the applicability of the Hertz model to the experimental data. After analyzing 180 curves, we obtained an average value of Young's modulus for the LOx layer in the 0.5-0.8 GPa range. These results agreed with those obtained for LOx submonolayer deposits on mica substrates, which allows discarding any important contribution from the underlying substrate on the measured properties. This range of values is closer to those obtained by other techniques on other globular proteins in comparison with those reported in previous AFM studies on similar systems. We found that for our experimental conditions the force curves can be, in principle, well fitted by the Hertz model for both conical and spherical indenter geometries. However, as the Young's modulus obtained for both geometries can differ appreciably, it becomes necessary to assess which indenter geometry is more adequate to explain the experimental data. For such purpose a systematic study of the indentation versus applied force curves obtained from both fittings for all the experimental curves was done.

  17. Lactation

    PubMed Central

    1989-01-01

    period when the use of artificial teats and dummies (pacifiers) may condition the infant to different oral actions that are inappropriate for breast-feeding. Comparisons of breast milk and cow's milk fail to describe the many important differences between them, e.g., the structural and qualitative differences in proteins and fats, and the bioavailability of minerals. The protection against infection and allergies conferred on the infant, which is impossible to attain through any other feeding regimen, is one of breast milk's most outstanding qualities. The maximum birth-spacing effect of lactation is achieved when an infant is fully, or nearly fully, breast-fed and the mother consequently remains amenorrhoeic. PMID:20604468

  18. PTHrP drives breast tumor initiation, progression, and metastasis in mice and is a potential therapy target

    PubMed Central

    Li, Jiarong; Karaplis, Andrew C.; Huang, Dao C.; Siegel, Peter M.; Camirand, Anne; Yang, Xian Fang; Muller, William J.; Kremer, Richard

    2011-01-01

    Parathyroid hormone–related protein (PTHrP) is a secreted factor expressed in almost all normal fetal and adult tissues. It is involved in a wide range of developmental and physiological processes, including serum calcium regulation. PTHrP is also associated with the progression of skeletal metastases, and its dysregulated expression in advanced cancers causes malignancy-associated hypercalcemia. Although PTHrP is frequently expressed by breast tumors and other solid cancers, its effects on tumor progression are unclear. Here, we demonstrate in mice pleiotropic involvement of PTHrP in key steps of breast cancer — it influences the initiation and progression of primary tumors and metastases. Pthrp ablation in the mammary epithelium of the PyMT-MMTV breast cancer mouse model caused a delay in primary tumor initiation, inhibited tumor progression, and reduced metastasis to distal sites. Mechanistically, it reduced expression of molecular markers of cell proliferation (Ki67) and angiogenesis (factor VIII), antiapoptotic factor Bcl-2, cell-cycle progression regulator cyclin D1, and survival factor AKT1. PTHrP also influenced expression of the adhesion factor CXCR4, and coexpression of PTHrP and CXCR4 was crucial for metastatic spread. Importantly, PTHrP-specific neutralizing antibodies slowed the progression and metastasis of human breast cancer xenografts. Our data identify what we believe to be new functions for PTHrP in several key steps of breast cancer and suggest that PTHrP may constitute a novel target for therapeutic intervention. PMID:22056386

  19. Lactation-induced cadmium-binding proteins

    SciTech Connect

    Bhattacharyya, M.H.; Solaiman, D.; Garvey, J.S.; Miyazaki, W.Y.

    1987-01-01

    Previously we have demonstrated an increase during midlactation in /sup 109/Cd adsorption and increased retention by the duodenum, kidney, and mammary tissue of mouse dams receiving environmental levels of cadmium//sup 109/Cd via drinking water, with little change in /sup 109/Cd retention in liver and jejunum compared to nonpregnant controls. Results are reported here of a study of cadmium deposition during midlactation as associated with induction of metallothionein (MT). A cadmium/hemoglobin (Cd/Hb) assay and radioimmunoassay for MT which measures heat-stable cadmium binding capacity in tissues was used to determine MT concentrations in fractions of kidney, liver, duodenum, and jejunum from female mice. Both assays demonstrated clear lactation-induced increases in MT concentrations in liver, kidney, and duodenum, with MT concentrations falling rapidly to control levels after weaning. 4 refs., 1 tab.

  20. [Bone loss in lactating women and post-pregnancy osteoporosis].

    PubMed

    Hirata, Go; Chaki, Osamu

    2011-09-01

    Measurement of the bone mineral density have shown that lactating women had 1 to 3% decrease in bone mineral density. Post pregnancy osteoporosis is rare condition that causes fragile fracture mostly in vertebrae. The bone loss in lactating women is caused by calcium loss, decrease in estrogen level, and increase in PTHrP (parathyroid hormone related protein) level. Some data have shown that extended lactation and amenorrhea had an association with the degree of bone loss. Mostly, the bone loss of the lactating women recovers to the baseline level, soon after the weaning, and there is no long term effect. Post pregnancy osteoporosis should be concerned, when we see a lactating woman with fragile fracture of the vertebrae.

  1. RSK activation via ERK modulates human colon cancer cells response to PTHrP.

    PubMed

    Calvo, Natalia; Carriere, Pedro; Martin, Maria Julia; Gentili, Claudia

    2017-04-06

    Parathyroid Hormone-related Peptide (PTHrP) is associated with several human cancers such as colon carcinoma. This disease is a complex multistep process that involves enhanced cell cycle progression and migration. Recently we obtained evidence that in the human colorectal adenocarcinoma Caco-2 cells, exogenous PTHrP increases the proliferation and positively modulates cell cycle progression via ERK1/2, p38 MAPK and PI3K. The purpose of this study was to explore if the serine/threonine kinase RSK, which is involved in the progress of many cancers and it is emerging as a potential therapeutic target, mediates PTHrP effects on cancer colon cells. Western blot analysis revealed that PTHrP increases RSK phosphorylation via ERK1/2 signaling pathway but not through p38 MAPK. Performing subcellular fractionation we found that the peptide also induces the nuclear localization of activated RSK, where many of its substrates are located. RSK participates in cell proliferation, in the up-regulation of cyclin D1 and CDK6 and in the down-regulation of p53 induced by PTHrP. Wound healing and transwell filters assays revealed that cell migration increased after PTHrP treatment. In addition, the hormone increases the protein expression of the focal adhesion kinase FAK, a regulator of cell motility. We observed that PTHrP induces cell migration and modulates FAK protein expression through ERK/ RSK signaling pathway but not via p38 MAPK pathway. Finally, in vivo studies revealed that the hormone activates RSK in xenografts tumor. Taken together, our findings provide new insights into the deregulated cell cycle and migration that is characteristic of tumor intestinal cells.

  2. Lactational evaluation of protein supplements of varying ruminal degradabilities.

    PubMed

    Henson, J E; Schingoethe, D J; Maiga, H A

    1997-02-01

    Twelve lactating Holstein cows (9 multiparous and 3 primiparous) were used in a replicated 3 x 3 Latin square design with three periods of 4 wk each to evaluate diets containing three protein supplements that varied in ruminally undegradable protein and amino acid (AA) composition. Diets contained either 44% crude protein (CP) solvent-extracted soybean meal, expeller (mechanically extracted) soybean meal, or a blend of animal and vegetable proteins as the protein supplement. The animal and vegetable blend consisted of equal portions of protein from blood meal, corn gluten meal, meat and bone meal, and soybean meal. All diets contained 33.3% alfalfa haylage, 16.7% corn silage, and 50% of the respective concentrate mix (dry matter basis). Diets contained 17.4, 17.8, and 17.8% CP and 34, 45, and 45% of CP as ruminally undegradable protein, respectively. Dry matter intake, milk production and composition, and body weight were similar among treatments. Uptakes of AA by the mammary gland were similar among treatments. The apparent first-limiting AA for each diet was likely Met, but Lys and Phe were also potentially limiting. Varying degrees of protein degradability and AA composition within the range of this study did not affect lactational responses, indicating that all of these protein supplements were adequate to support milk production.

  3. Regulation of PTHrP expression by cyclic mechanical strain in postnatal growth plate chondrocytes.

    PubMed

    Xu, Tao; Yang, Kaixiang; You, Hongbo; Chen, Anmin; Wang, Jiang; Xu, Kai; Gong, Chen; Shao, Jingfan; Ma, Zhongxi; Guo, Fengjing; Qi, Jun

    2013-10-01

    Mechanical loading has been widely considered to be a crucial regulatory factor for growth plate development, but the exact mechanisms of this regulation are still not completely understood. In the growth plate, parathyroid hormone-related protein (PTHrP) regulates chondrocyte differentiation and longitudinal growth. Cyclic mechanical strain has been demonstrated to influence growth plate chondrocyte differentiation and metabolism, whereas the relationship between cyclic mechanical strain and PTHrP expression is not clear. The objective of this study was to investigate whether short-term cyclic tensile strain regulates PTHrP expression in postnatal growth plate chondrocytes in vitro and to explore whether the organization of cytoskeletal F-actin microfilaments is involved in this process. To this end, we obtained growth plate chondrocytes from 2-week-old Sprague-Dawley rats and sorted prehypertrophic and hypertrophic chondrocytes using immunomagnetic beads coated with anti-CD200 antibody. The sorted chondrocytes were subjected to cyclic tensile strain of varying magnitude and duration at a frequency of 0.5 Hz. We found that cyclic strain regulates PTHrP expression in a magnitude- and time-dependent manner. Incubation of chondrocytes with cytochalasin D, an actin microfilament-disrupting reagent, blocked the induction of PTHrP expression in response to strain. The results suggest that short-term cyclic tensile strain induces PTHrP expression in postnatal growth plate prehypertrophic and hypertrophic chondrocytes and that PTHrP expression by these chondrocytes may subsequently affect growth plate development. The results also support the idea that the organization of cytoskeletal F-actin microfilaments plays an important role in mechanotransduction.

  4. PTHrP Interacts With the TGF-β/BMP-2/Gremlin Signaling Pathway to Regulate Pro-inflammatory and Pro-fibrotic Mediators in Pancreatic Acinar and Stellate Cells

    PubMed Central

    Bhatia, Vandanajay; Cao, Yanna; Ko, Tien C.; Falzon, Miriam

    2015-01-01

    Objectives TGF-β regulates immune and fibrotic responses of chronic pancreatitis (CP). The bone morphogenetic protein-2 (BMP-2) antagonist gremlin is regulated by TGF-β. Parathyroid hormone-related hormone (PTHrP) levels are elevated in CP. Here we investigated the crosstalk between TGF-β/BMP-2/gremlin and PTHrP signaling. Methods Reverse transcription/real-time PCR, ChIP, Western blotting, and transient transfection were used to investigate PTHrP regulation by TGF-β and BMP-2, and gremlin regulation by PTHrP. The PTHrP antagonist PTHrP (7-34) and acinar cells with conditional Pthrp gene deletion (PTHrPΔacinar) were used to assess PTHrP’s role in the pro-inflammatory and pro-fibrotic effects of TGF-β and gremlin. Results TGF-β increased PTHrP levels in acinar cells and pancreatic stellate cells (PSCs) through a Smad3-dependent pathway. TGF-β’s effects on levels of IL-6 and ICAM-1(acinar cells) and procollagen I and fibronectin (PSCs) were inhibited by PTHrP (7-34). PTHrPΔacinar suppressed TGF-β’s effects on IL-6 and ICAM-1. PTHrP increased gremlin in acinar cells, and inhibiting gremlin action suppressed TGF-β’s and PTHrP’s effects on IL-6 and ICAM-1. TGF-β-mediated gremlin upregulation was suppressed in PTHrPΔacinar cells. BMP-2 suppressed PTHrP levels in PSCs. Conclusions PTHrP functions as a novel mediator of the pro-inflammatory and pro-fibrotic effects of TGF-β. TGF-β and BMP-2 regulate PTHrP expression and PTHrP regulates gremlin levels. PMID:26495794

  5. Trps1 deficiency enlarges the proliferative zone of growth plate cartilage by upregulation of Pthrp.

    PubMed

    Nishioka, Katsuhiro; Itoh, Shunji; Suemoto, Hiroki; Kanno, Seiji; Gai, Zhibo; Kawakatsu, Motohisa; Tanishima, Hiroyuki; Morimoto, Yoshifumi; Hatamura, Ikuji; Yoshida, Munehito; Muragaki, Yasuteru

    2008-07-01

    We have reported that elongation of the columnar proliferative zone of long bone growth plates in Trps1-/- mice during the late fetal stage in the previous study [1]. Since expression of Trps1 protein was found to overlap with that of mRNAs for Indian hedgehog (Ihh), PTH/PTHrP receptor (PPR), and PTHrP, we hypothesized that Trps1 may inhibit the hypertrophic differentiation of chondrocytes by interacting with the Ihh/PTHrP feedback loop. To investigate whether Trps1 has a role in this Ihh/PTHrP feedback loop, we compared the growth plates of Trps1-/- mice and wild-type (Trps1+/+) mice. Immunohistochemistry showed that Trps1 protein was strongly expressed in the periarticular and prehypertrophic zones of the fetal growth plate in wild-type mice on embryonic day 18.5 (E18.5). On the other hand, Ihh, PPR, and PTHrP mRNAs were predominantly expressed in the prehypertrophic zone at this stage of development. While expression of Ihh and PPR by prehypertrophic chondrocytes was unaffected in the growth plates of Trps1-/- mice, the range of PTHrP expression was expanded toward the proliferating zone in these mice. Quantitative real-time PCR analysis demonstrated upregulation of PTHrP in the epiphyseal growth plates of Trps1-/- mice. Furthermore, promoter analysis combined with the chromatin immunoprecipitation (ChIP) assay demonstrated that direct binding of Trps1 to the PTHrP promoter suppressed the transcription of PTHrP. Finally, organ culture of E14.5 tibiae in the absence or the presence of Pthrp revealed that the proliferative zone of the tibial growth plate was elongated by culture with Pthrp compared to that of control tibiae. Taken together, these data provide the first genetic evidence that lack of Trps1 leads to overexpression of PTHrP, and that Trps1 is required to maintain the normal organization of chondrocytes in the growth plate.

  6. PTHrP produced by myeloma plasma cells regulates their survival and pro-osteoclast activity for bone disease progression.

    PubMed

    Cafforio, Paola; Savonarola, Annalisa; Stucci, Stefania; De Matteo, Monica; Tucci, Marco; Brunetti, Anna Elisabetta; Vecchio, Vita Mariagrazia; Silvestris, Francesco

    2014-01-01

    To promote their survival and progression in the skeleton, osteotropic malignancies of breast, lung, and prostate produce parathyroid hormone-related protein (PTHrP), which induces hypercalcemia. PTHrP serum elevations have also been described in multiple myeloma (MM), although their role is not well defined. When we investigated MM cells from patients and cell lines, we found that PTHrP and its receptor (PTH-R1) are highly expressed, and that PTHrP is secreted both as a full-length molecule and as small subunits. Among these subunits, the mid-region, including the nuclear localization sequence (NLS), exerted a proliferative effect because it was accumulated in nuclei of MM cells surviving in starvation conditions. This was confirmed by increased transcription of several genes enrolled in proliferation and apoptosis control. PTHrP was also found to stimulate PTH-R1 in MM cells. PTH-R1's selective activation by the full-length PTHrP molecule or the NH2 -terminal fragment resulted in a significant increase of intracellular Ca(2+) influx, cyclic adenosine monophosphate (cAMP) content, and expression of receptor activator of NF-κB ligand (RANKL) and monocyte chemoattractant protein-1 (MCP-1). Our data definitely clarify the role of PTHrP in MM. The PTHrP peptide is functionally secreted by malignant plasma cells and contributes to MM tumor biology and progression, both by intracrine maintenance of cell proliferation in stress conditions and by autocrine or paracrine stimulation of PTH-R1, which in turn reinforces the production of osteoclastogenic factors. © 2014 American Society for Bone and Mineral Research. © 2014 American Society for Bone and Mineral Research.

  7. The midregion, nuclear localization sequence, and C terminus of PTHrP regulate skeletal development, hematopoiesis, and survival in mice

    PubMed Central

    Toribio, Ramiro E.; Brown, Holly A.; Novince, Chad M.; Marlow, Brandlyn; Hernon, Krista; Lanigan, Lisa G.; Hildreth, Blake E.; Werbeck, Jillian L.; Shu, Sherry T.; Lorch, Gwendolen; Carlton, Michelle; Foley, John; Boyaka, Prosper; McCauley, Laurie K.; Rosol, Thomas J.

    2010-01-01

    The functions of parathyroid hormone-related protein (PTHrP) on morphogenesis, cell proliferation, apoptosis, and calcium homeostasis have been attributed to its N terminus. Evidence suggests that many of these effects are not mediated by the N terminus but by the midregion, a nuclear localization sequence (NLS), and C terminus of the protein. A knock-in mouse lacking the midregion, NLS, and C terminus of PTHrP (PthrpΔ/Δ) was developed. PthrpΔ/Δ mice had craniofacial dysplasia, chondrodysplasia, and kyphosis, with most mice dying by d 5 of age. In bone, there were fewer chondrocytes and osteoblasts per area, bone mass was decreased, and the marrow was less cellular, with erythroid hypoplasia. Cellular proliferation was impaired, and apoptosis was increased. Runx2, Ocn, Sox9, Crtl1, β-catenin, Runx1, ephrin B2, cyclin D1, and Gata1 were underexpressed while P16/Ink4a, P21, GSK-3β, Il-6, Ffg3, and Ihh were overexpressed. Mammary gland development was aberrant, and energy metabolism was deregulated. These results establish that the midregion, NLS, and C terminus of PTHrP are crucial for the commitment of osteogenic and hematopoietic precursors to their lineages, and for survival, and many of the effects of PTHrP on development are not mediated by its N terminus. The down-regulation of Runx1, Runx2, and Sox9 indicates that PTHrP is a modulator of transcriptional activation during stem cell commitment. Toribio, R. E., Brown, H. A., Novince, C. M., Marlow, B. Hernon, K., Lanigan, L. G., Hildreth III, B. E., Werbeck, J. L., Shu, S. T., Lorch, G., Carlton, M., Foley, J., Boyaka, P., McCauley, L. K., Rosol, T. J. The midregion, nuclear localization sequence, and C terminus of PTHrP regulate skeletal development, hematopoiesis, and survival in mice. PMID:20145205

  8. Integrin β4 regulation of PTHrP underlies its contribution to mammary gland development.

    PubMed

    Li, Jiarong; Sun, Huayan; Feltri, M Laura; Mercurio, Arthur M

    2015-11-15

    The integrin α6β4 (referred to as β4) is expressed in epithelial cells where it functions as a laminin receptor. Although in vitro studies have implicated β4 in the biology of mammary epithelial cells, its contribution to mammary gland development has not been settled. To address this problem, we generated and analyzed itgb4(flox/flox)MMTV-Cre(-) and itgb4(flox/flox)MMTV-Cre(+) mice. The salient features of embryonic mammary tissue from itgb4(flox/flox)MMTV-Cre(+) mice were significantly smaller mammary buds and increased apoptosis in the surrounding mesenchyme. Also, compared to control glands, the itgb4-deleted mammary buds lacked expression of the progenitor cell marker CK14 and they were unable to generate mammary glands upon transplantation into cleared fat pads of recipient mice. Analysis of mammary glands at puberty and during pregnancy revealed that itgb4-diminished mammary tissue was unable to elongate and undergo branching morphogenesis. Micro-dissection of epithelial cells in the mammary bud and of the surrounding mesenchyme revealed that loss of β4 resulted in a significant decrease in the expression of parathyroid hormone related protein (PTHrP) in epithelial cells and of target genes of the PTHrP receptor in mesenchymal cells. Given that the phenotype of the itgb4-deleted mammary tissue mimicked that of the PTHrP knockout, we hypothesized that β4 contributes to mammary gland development by sustaining PTHrP expression and enabling PTHrP signaling. Indeed, the inability of itgb4-deleted mammary buds to elongate was rescued by exogenous PTHrP. These data implicate a critical role for the β4 integrin in mammary gland development and provide a mechanism for this role.

  9. Amino acid metabolism and protein synthesis in lactating rats fed on a liquid diet.

    PubMed Central

    Barber, T; García de la Asunción, J; Puertes, I R; Viña, J R

    1990-01-01

    1. Amino acid metabolism was studied in control virgin rats, lactating rats and virgin rats protein-pair-fed with the lactating rats (high-protein virgin rats). 2. Urinary excretion of nitrogen and urea was higher in lactating than in control virgin rats, and in high-protein virgin rats it was higher than in lactating rats. 3. The activities of urea-cycle enzymes (units/g) were higher in high-protein virgin than in lactating rats, except for arginase. In lactating rats the activities of carbamoyl-phosphate synthase, ornithine carbamoyltransferase and argininosuccinate synthase were lower than in control virgin rats. When the liver size is considered, the activities in lactating rats were similar to those in high-protein virgin rats, except for arginase. 4. N-Acetylglutamate content was higher in high-protein virgin rats than in the other two groups. 5. The rate of urea synthesis from precursors by isolated hepatocytes was higher in high-protein virgin rats than in the other two groups. 6. The flooding-dose method (L-[4-3H]phenylalanine) for measuring protein synthesis was used. The absolute synthesis rates of mammary gland, liver and small-intestinal mucosa were higher in lactating rats than in the other two groups, and in high-protein virgin rats than in control virgin rats 7. These results show that the increased needs for amino acids during lactation are met by hyperphagia and by a nitrogen-sparing mechanism. PMID:2396994

  10. Endocrine regulation of carbonate precipitate formation in marine fish intestine by stanniocalcin and PTHrP.

    PubMed

    Gregório, Sílvia F; Carvalho, Edison S M; Campinho, Marco A; Power, Deborah M; Canário, Adelino V M; Fuentes, Juan

    2014-05-01

    In marine fish, high epithelial bicarbonate secretion by the intestine generates luminal carbonate precipitates of divalent cations that play a key role in water and ion homeostasis. In vitro studies highlight the involvement of the calciotropic hormones PTHrP (parathyroid hormone-related protein) and stanniocalcin (STC) in the regulation of epithelial bicarbonate transport. The present study tested the hypothesis that calciotropic hormones have a regulatory role in carbonate precipitate formation in vivo. Sea bream (Sparus aurata) juveniles received single intraperitoneal injections of piscine PTHrP(1-34), the PTH/PTHrP receptor antagonist PTHrP(7-34) or purified sea bream STC, or were passively immunized with polyclonal rabbit antisera raised against sea bream STC (STC-Ab). Endocrine effects on the expression of the basolateral sodium bicarbonate co-transporter (Slc4a4.A), the apical anion exchangers Slc26a6.A and Slc26a3.B, and the V-type proton pump β-subunit (Atp6v1b) in the anterior intestine were evaluated. In keeping with their calciotropic nature, the hypocalcaemic factors PTHrP(7-34) and STC up-regulated gene expression of all transporters. In contrast, the hypercalcaemic factor PTHrP(1-34) and STC antibodies down-regulated transporters involved in the bicarbonate secretion cascade. Changes in intestine luminal precipitate contents provoked by calcaemic endocrine factors validated these results: 24 h post-injection either PTHrP(1-34) or immunization with STC-Ab reduced the carbonate precipitate content in the sea bream intestine. In contrast, the PTH/PTHrP receptor antagonist PTHrP(7-34) increased not only the precipitated fraction but also the concentration of HCO3(-) equivalents in the intestinal fluid. These results confirm the hypothesis that calciotropic hormones have a regulatory role in carbonate precipitate formation in vivo in the intestine of marine fish. Furthermore, they illustrate for the first time in fish the counteracting effect of PTHrP

  11. Signal transducer and activator of transcription 5a inhibited by pimozide may regulate survival of goat mammary gland epithelial cells by regulating parathyroid hormone-related protein.

    PubMed

    Li, Hui; Zheng, Huiling; Sun, Yongsen; Yu, Qian; Li, Lihui

    2014-11-10

    The signal transducer and activator of transcription 5a (Stat5a) modulates genes involved in proliferation and survival and plays pivotal roles in regulating the function of the mammary gland during pregnancy, lactation, and involution. However, there is little information about the effects of Stat5a on apoptosis of goat mammary gland epithelial cells (GMECs). In addition, parathyroid hormone-related protein (PTHrP) is a key regulator in cellular calcium transport, mammary gland development and breast tumor biology. This study aimed to explore the interaction of Stat5a and PTHrP in GMEC apoptosis. Quantitative real time PCR (qRT-PCR) suggested that Stat5a was predominantly expressed in the mammary gland, lung, liver and spleen of goats. Treating the GMECs with pimozide, an inhibitor of Stat5a that decreases Stat5a tyrosine phosphorylation, increased PTHrP levels in GMECs in a dose-dependent manner and simultaneously promoted apoptosis of the GMECs. We also demonstrated that PTHrP inhibition induced GMEC apoptosis and restrained cell proliferation. In contrast, PTHrP overexpression protected GMECs from pimozide- and calcium-induced apoptosis, and promoted cell proliferation. Furthermore, pimozide and CaCl2 downregulated the antiapoptotic protein Bcl-2 mRNA expression, respectively, and these effects were protected by PTHrP overexpression. Interestingly, we also found that Stat5a suppressed the expression of matrix metalloproteinase 9 (MMP-9) which can induce goat mammary epithelial cell migration, but PTHrP increased MMP-9 mRNA level. Thus, Stat5a may regulate GMEC survival by regulating the expression of PTHrP.

  12. Low-Protein Diet during Lactation and Maternal Metabolism in Rats

    PubMed Central

    Moretto, Vera L.; Ballen, Marcia O.; Gonçalves, Talita S. S.; Kawashita, Nair H.; Stoppiglia, Luiz F.; Veloso, Roberto V.; Latorraca, Márcia Q.; Martins, Maria Salete F.; Gomes-da-Silva, Maria Helena G.

    2011-01-01

    Some metabolic alterations were evaluated in Wistar rats which received control or low-protein (17%; 6%) diets, from the pregnancy until the end of lactation: control non-lactating (CNL), lactating (CL), low-protein non-lactating (LPNL) and lactating (LPL) groups. Despite the increased food intake by LPL dams, both LP groups reduced protein intake and final body mass was lower in LPL. Higher serum glucose occurred in both LP groups. Lactation induced lower insulin and glucagon levels, but these were reduced by LP diet. Prolactin levels rose in lactating, but were impaired in LPL, followed by losses of mammary gland (MAG) mass and, a fall in serum leptin in lactating dams. Lipid content also reduced in MAG and gonadal white adipose tissue of lactating and, in LPL, contributed to a decreased daily milk production, and consequent impairment of body mass gain by LPL pups. Liver mass, lipid content and ATP-citrate enzyme activity were increased by lactation, but malic enzyme and lipid: glycogen ratio elevated only in LPL. Conclusion. LP diet reduced the development of MAG and prolactin secretion which compromised milk production and pups growth. Moreover, this diet enhanced the store of lipid to glycogen ratio and suggests a higher risk of fatty liver development. PMID:21637364

  13. Associations between milk protein concentration at various stages of lactation and reproductive performance in dairy cows.

    PubMed

    Morton, J M; Auldist, M J; Douglas, M L; Macmillan, K L

    2016-12-01

    Milk protein concentration has been positively associated with a range of measures of reproductive performance in dairy cows. These beneficial associations are most likely due to factors affecting both milk protein concentration and reproductive performance possibly being mediated, in part, by energy balance during early lactation. However, it is likely that factors other than energy balance are also involved in these relationships. A retrospective single cohort study was conducted using subsets of data collected from 74 dairy herds with seasonal or split calving patterns. Associations between milk protein concentration at various stages of lactation and reproductive performance in Holstein dairy cows were assessed using random effects logistic regression and survival analysis with milk protein concentration during the cow's breeding period fitted as a time-varying covariate. The beneficial associations between milk protein concentration and each of the 4 selected indices for measuring reproductive performance were evident when milk protein concentration was derived for each 30-d period from calving up to 300d in milk. For the first 150d of lactation the adjusted odds ratios were highest from 31 to 60d and only slightly lower for all periods up to 150d of lactation. Estimated associations for 31 to 60d were stronger than for 0 to 30d. In addition, milk protein concentration during a cow's breeding period was positively associated with the subsequent daily hazard of conception, even after adjusting for milk protein concentration in the cow's first or second month of lactation. Milk protein concentrations from 0 to 30d of lactation were less closely correlated with concentrations measured at subsequent 30-d intervals; correlations were closer between other periods in lactation. These results indicate that the association between milk protein concentration and reproductive performance is partly due to factors other than the extent of negative energy balance in early

  14. Interaction of protein percent with caloric density and protein source for lactating cows.

    PubMed

    Van Horn, H H; Blanco, O; Harris, B; Beede, D K

    1985-07-01

    Experiment 1 was to test effect of three ratios of energy to protein in complete mixed diets for 36 lactating cows in three, 28-d periods. Energy was varied with cottonseed hulls, pelleted ground corrugated boxes, and a mixture of the two. Crude protein was varied with soybean meal to give energy:crude protein of 5.7, 5.0, and 4.6 for each energy amount. Cottonseed meal was compared with soybean meal in corrugated box diets. Feed intake was much higher with cottonseed hulls, and appreciable feedlot bloat resulted from pelleted ground corrugated box diets. Data adjusted to equal feed intake showed significant effect of energy to crude protein ratio on milk yield and improved digestion of organic matter with soybean meal vs. cottonseed meal. Experiment 2 tested the hypothesis that lactating cows consuming high-protein alfalfa may benefit from supplemental protein. Diets were 50% forage. Six diets were 14 or 18% crude protein in three ratios of alfalfa hay to corn silage (0:100, 50:50, 100:0). Additional corn silage diets were to compare: 14 versus 18% protein from distiller's dried grains with solubles only and with .5 or .9% urea (four diets); two 14% protein diets compared .6% added potassium chloride with or without .5% urea. Thirty-six Holstein cows in early lactation received one of the 12 diets in each of three 28-d periods. Distiller's grains with solubles markedly depressed milk yield (2.2 kg/d) and milk protein (.22%); heat damage of distiller's grains was evident. Protein interacted with alfalfa so gain in milk from 18 versus 14% increased from .55 to 1.36 to 2.66 kg/d as alfalfa changed from 0 to 50 to 100%. Thus, crude protein of alfalfa was not as effective as that from soybean meal in supporting milk yield.

  15. Parathyroid hormone-related peptide in lactation and in umbilical cord blood.

    PubMed

    Khosla, S; Johansen, K L; Ory, S J; O'Brien, P C; Kao, P C

    1990-11-01

    Parathyroid hormone-related peptide (PTHrP) is expressed in lactating rat mammary glands after suckling, as a result of increases in prolactin rather than suckling per se. In addition, PTHrP produced in the fetal parathyroid glands and placenta may be responsible for stimulation of placental calcium transport. In the current study, we used a radioimmunoassay for human PTHrP to measure levels of the peptide in (1) human breast milk, cow's milk, and two infant formulas; (2) sequential plasma samples in prepartum and postpartum lactating women; (3) women with pathologic hyperprolactinemia; and (4) human umbilical cord blood. In normal subjects, plasma PTHrP levels ranged from less than 2 to 5 pmol/liter. In contrast, human breast milk contained substantially increased levels of immunoreactive PTHrP. Similar elevations were found in cow's milk and in one infant formula. Column chromatography of breast milk demonstrated that PTHrP immunoreactivity included a region of adenylate cyclase stimulating activity, consistent with the presence of biologically active PTHrP. Plasma prepartum PTHrP values did not differ from corresponding postpartum values in lactating women. Women with hyperprolactinemia had a mean plasma PTHrP value in the high-normal range. Umbilical cord blood had considerably suppressed parathyroid hormone values but PTHrP levels that were indistinguishable from those in normal human plasma. Thus, PTHrP is present in high concentrations in breast milk but apparently does not gain access to the maternal circulation in significant amounts. In addition, women with pathologic hyperprolactinemia seem not to have increased levels of circulating PTHrP.(ABSTRACT TRUNCATED AT 250 WORDS)

  16. PTHrP regulates water absorption and aquaporin expression in the intestine of the marine sea bream (Sparus aurata, L.).

    PubMed

    Carvalho, Edison S M; Gregório, Sílvia F; Canário, Adelino V M; Power, Deborah M; Fuentes, Juan

    2015-03-01

    Water ingestion by drinking is fundamental for ion homeostasis in marine fish. However, the fluid ingested requires processing to allow net water absorption in the intestine. The formation of luminal carbonate aggregates impacts on calcium homeostasis and requires epithelial HCO3(-) secretion to enable water absorption. In light of its endocrine importance in calcium handling and the indication of involvement in HCO3(-) secretion the present study was designed to expose the role of the parathyroid hormone-related protein (PTHrP) in HCO3(-) secretion, water absorption and the regulation of aqp1 gene expression in the anterior intestine of the sea bream. HCO3(-) secretion rapidly decreased when PTHrP(1-34) was added to anterior intestine of the sea bream mounted in Ussing chambers. The effect achieved a maximum inhibition of 60% of basal secretion rates, showing a threshold effective dose of 0.1 ng ml(-1) compatible with reported plasma values of PTHrP. When applied in combination with the adenylate cyclase inhibitor (SQ 22.536, 100 μmol l(-1)) or the phospholipase C inhibitor (U73122, 10 μmol l(-1)) the effect of PTHrP(1-34) on HCO3(-) secretion was reduced by about 50% in both cases. In parallel, bulk water absorption measured in intestinal sacs was sensitive to inhibition by PTHrP. The inhibitory action conforms to a typical dose-response curve in the range of 0.1-1000 ng ml(-1), achieves a maximal effect of 60-65% inhibition from basal rates and shows threshold significant effects at hormone levels of 0.1 ng ml(-1). The action of PTHrP in water absorption was completely abolished in the presence of the adenylate cyclase inhibitor (SQ 22.536, 100 μmol l(-1)) and was insensitive to the phospholipase C inhibitor (U73122, 10 μmol l(-1)). In vivo injections of PTHrP(1-34) or the PTH/PTHrP receptor antagonist PTHrP(7-34) evoked respectively, a significant decrease or increase of aqp1ab, but not aqp1a. Overall the present results suggest that PTHrP acts as a key

  17. Proteomic characterization of human milk fat globule membrane proteins during a 12 month lactation period.

    PubMed

    Liao, Yalin; Alvarado, Rudy; Phinney, Brett; Lönnerdal, Bo

    2011-08-05

    The milk fat globule membrane (MFGM) contains proteins which have been implicated in a variety of health benefits. Milk fat globule membrane proteins were isolated from human milk during a 12 month lactation period and subjected to in-solution digestion and liquid chromatography tandem mass spectrometry analysis. Data were pooled, and our results showed that 191 proteins were identified. Relative quantification of the identified MFGM proteins during the course of lactation was performed by label free spectral counting and differentiation expression analysis, which showed some proteins decreasing during the course of lactation whereas some increased or remained at a relatively constant level. The human MFGM proteins are distributed between intracellular, extracellular, and membrane-associated proteins, and they are mainly involved in cell communication and signal transduction, immune function, metabolism and energy production. This study provides more insights into the dynamic composition of human MFGM proteins, which in turn will enhance our understanding of the physiological significance of MFGM proteins.

  18. Lactation performance of mid-lactation dairy cows fed ruminally degradable protein at concentrations lower than national research council recommendations.

    PubMed

    Cyriac, J; Rius, A G; McGilliard, M L; Pearson, R E; Bequette, B J; Hanigan, M D

    2008-12-01

    The aim of this study was to test whether feeding of diets containing lower proportions of ruminally degradable protein (RDP) but with a constant proportion of ruminally undegradable protein (RUP) alters feed intake, milk production and yield, and the apparent efficiency of N utilization by mid-lactation dairy cows. During the covariate period (d 1 to 28), 40 mid-lactation cows (36 Holstein and 4 Jersey x Holstein cross-breds) were fed a common diet formulated to contain 11.3% of diet dry matter (DM) as RDP. During the treatment period (d 29 to 47), cows were randomly assigned to 1 of 4 diets formulated to contain 11.3, 10.1, 8.8, or 7.6% RDP, whereas ruminally undegradable protein remained constant at 7.1% of DM. All diets contained 47.5% forage and 52.5% concentrate on a DM basis. Dry matter intake was significantly reduced for the 7.6% RDP diet. The lowest RDP content was associated with a trend for reduced milk yield. Dietary RDP had no effect on body weight or milk fat, protein, and lactose contents. Milk protein yield was not affected by RDP level; however, milk fat yield decreased linearly as dietary RDP was reduced. Concentrations of plasma essential amino acids were unaffected, whereas milk urea-N concentrations decreased linearly as dietary RDP content was reduced. The apparent efficiency of N utilization for milk N production increased from 27.7% on the 11.3% RDP diet to 38.6% on the 7.6% RDP diet. The dietary RDP requirement of cows in this study was apparently met between 15.9 and 14.7% dietary crude protein. Milk production was not significantly affected by the 8.8% RDP (15.9% crude protein) diet even though the NRC (2001) model predicted that RDP supply was 87% of that required, suggesting the current NRC recommendations for RDP may be overestimated for mid-lactation dairy cows in this study.

  19. Automated High Throughput Protein Crystallization Screening at Nanoliter Scale and Protein Structural Study on Lactate Dehydrogenase

    SciTech Connect

    Li, Fenglei

    2006-08-09

    The purposes of our research were: (1) To develop an economical, easy to use, automated, high throughput system for large scale protein crystallization screening. (2) To develop a new protein crystallization method with high screening efficiency, low protein consumption and complete compatibility with high throughput screening system. (3) To determine the structure of lactate dehydrogenase complexed with NADH by x-ray protein crystallography to study its inherent structural properties. Firstly, we demonstrated large scale protein crystallization screening can be performed in a high throughput manner with low cost, easy operation. The overall system integrates liquid dispensing, crystallization and detection and serves as a whole solution to protein crystallization screening. The system can dispense protein and multiple different precipitants in nanoliter scale and in parallel. A new detection scheme, native fluorescence, has been developed in this system to form a two-detector system with a visible light detector for detecting protein crystallization screening results. This detection scheme has capability of eliminating common false positives by distinguishing protein crystals from inorganic crystals in a high throughput and non-destructive manner. The entire system from liquid dispensing, crystallization to crystal detection is essentially parallel, high throughput and compatible with automation. The system was successfully demonstrated by lysozyme crystallization screening. Secondly, we developed a new crystallization method with high screening efficiency, low protein consumption and compatibility with automation and high throughput. In this crystallization method, a gas permeable membrane is employed to achieve the gentle evaporation required by protein crystallization. Protein consumption is significantly reduced to nanoliter scale for each condition and thus permits exploring more conditions in a phase diagram for given amount of protein. In addition

  20. Proteomic characterization of human milk whey proteins during a twelve-month lactation period.

    PubMed

    Liao, Yalin; Alvarado, Rudy; Phinney, Brett; Lönnerdal, Bo

    2011-04-01

    Human milk is a rich source of bioactive proteins that support the early growth and development of the newborn. Although the major components of the protein fraction in human milk have been studied, the expression and relative abundance of minor components have received limited attention. We examined the expression of low-abundance proteins in the whey fraction of human milk and their dynamic changes over a twelve-month lactation period. The low-abundance proteins were enriched by ProteoMiner beads, and protein identification was performed by liquid chromatography tandem mass spectrometry. One hundred and fifteen proteins were identified, thirty-eight of which have not been previously reported in human colostrum or milk. We also for the first time described differences in protein patterns among the low-abundance proteins during lactation. These results enhance our knowledge about the complexity of the human milk proteome, which constitutes part of the advantages to the breast-fed infant.

  1. Green tea extract intake during lactation modified cardiac macrophage infiltration and AMP-activated protein kinase phosphorylation in weanling rats from undernourished mother during gestation and lactation.

    PubMed

    Matsumoto, E; Kataoka, S; Mukai, Y; Sato, M; Sato, S

    2017-04-01

    Maternal dietary restriction is often associated with cardiovascular disease in offspring. The aim of this study was to investigate the effect of green tea extract (GTE) intake during lactation on macrophage infiltration, and activation of adenosine monophosphate (AMP)-activated protein kinase (AMPK) and serine-threonine kinase Akt (Akt) in the hearts of weanlings exposed to maternal dietary protein restriction. Pregnant Wistar rats were fed control (C) or low-protein diets (LP) throughout gestation. Following delivery, the dams received a control or a GTE-containing control diet during lactation: control diet during gestation and lactation (CC), low-protein diet during gestation and lactation (LPC), low-protein diet during gestation and 0.12% GTE-containing low-protein diet during lactation (LPL), and low-protein diet during gestation and 0.24% GTE-containing low-protein diet during lactation (LPH). The female offspring were sacrificed at day 22. Biochemical parameters in the plasma, macrophage infiltration, degree of fibrosis and expression levels of AMPK and Akt were examined. The plasma insulin level increased in LPH compared with LPC. Percentage of the fibrotic areas and the number of macrophages in LPC were higher than those in CC. Conversely, the fibrotic areas and the macrophage number in LPH were smaller (21 and 56%, respectively) than those in LPC. The levels of phosphorylated AMPK in LPL and LPH, and Akt in LPH were greater than those in LPC. In conclusion, maternal protein restriction may induce macrophage infiltration and the decrease of insulin levels. However, GTE intake during lactation may suppress macrophage infiltration and restore insulin secretion function via upregulation of AMPK and insulin signaling in weanlings.

  2. Identification of lactate dehydrogenase as a mammalian pyrroloquinoline quinone (PQQ)-binding protein

    PubMed Central

    Akagawa, Mitsugu; Minematsu, Kenji; Shibata, Takahiro; Kondo, Tatsuhiko; Ishii, Takeshi; Uchida, Koji

    2016-01-01

    Pyrroloquinoline quinone (PQQ), a redox-active o-quinone, is an important nutrient involved in numerous physiological and biochemical processes in mammals. Despite such beneficial functions, the underlying molecular mechanisms remain to be established. In the present study, using PQQ-immobilized Sepharose beads as a probe, we examined the presence of protein(s) that are capable of binding PQQ in mouse NIH/3T3 fibroblasts and identified five cellular proteins, including l-lactate dehydrogenase (LDH) A chain, as potential mammalian PQQ-binding proteins. In vitro studies using a purified rabbit muscle LDH show that PQQ inhibits the formation of lactate from pyruvate in the presence of NADH (forward reaction), whereas it enhances the conversion of lactate to pyruvate in the presence of NAD+ (reverse reaction). The molecular mechanism underlying PQQ-mediated regulation of LDH activity is attributed to the oxidation of NADH to NAD+ by PQQ. Indeed, the PQQ-bound LDH oxidizes NADH, generating NAD+, and significantly catalyzes the conversion of lactate to pyruvate. Furthermore, PQQ attenuates cellular lactate release and increases intracellular ATP levels in the NIH/3T3 fibroblasts. Our results suggest that PQQ, modulating LDH activity to facilitate pyruvate formation through its redox-cycling activity, may be involved in the enhanced energy production via mitochondrial TCA cycle and oxidative phosphorylation. PMID:27230956

  3. Identification of lactate dehydrogenase as a mammalian pyrroloquinoline quinone (PQQ)-binding protein.

    PubMed

    Akagawa, Mitsugu; Minematsu, Kenji; Shibata, Takahiro; Kondo, Tatsuhiko; Ishii, Takeshi; Uchida, Koji

    2016-05-27

    Pyrroloquinoline quinone (PQQ), a redox-active o-quinone, is an important nutrient involved in numerous physiological and biochemical processes in mammals. Despite such beneficial functions, the underlying molecular mechanisms remain to be established. In the present study, using PQQ-immobilized Sepharose beads as a probe, we examined the presence of protein(s) that are capable of binding PQQ in mouse NIH/3T3 fibroblasts and identified five cellular proteins, including l-lactate dehydrogenase (LDH) A chain, as potential mammalian PQQ-binding proteins. In vitro studies using a purified rabbit muscle LDH show that PQQ inhibits the formation of lactate from pyruvate in the presence of NADH (forward reaction), whereas it enhances the conversion of lactate to pyruvate in the presence of NAD(+) (reverse reaction). The molecular mechanism underlying PQQ-mediated regulation of LDH activity is attributed to the oxidation of NADH to NAD(+) by PQQ. Indeed, the PQQ-bound LDH oxidizes NADH, generating NAD(+), and significantly catalyzes the conversion of lactate to pyruvate. Furthermore, PQQ attenuates cellular lactate release and increases intracellular ATP levels in the NIH/3T3 fibroblasts. Our results suggest that PQQ, modulating LDH activity to facilitate pyruvate formation through its redox-cycling activity, may be involved in the enhanced energy production via mitochondrial TCA cycle and oxidative phosphorylation.

  4. Human Milk Analyser shows that the lactation period affects protein levels in preterm breastmilk.

    PubMed

    Kreissl, Alexandra; Zwiauer, Valentina; Repa, Andreas; Binder, Christoph; Thanhaeuser, Margarita; Jilma, Bernd; Berger, Angelika; Haiden, Nadja

    2016-06-01

    This study measured the composition of preterm human breastmilk, particularly the protein content, with the MIRIS Human Milk Analyser, compared our results with published values and determined the relationship between protein content and lactation period. We analysed 83 samples of 24-hour pooled human milk from 76 mothers who delivered preterm infants weighing under 1500 g at less than 32 weeks of gestational age. The milk's protein, fat and energy were measured by the MIRIS Human Milk Analyser and compared to reference values. The relationship between protein content and lactation period was quantified. On average, the samples contained 1.1 ± 0.37 g (0.2-2.2 g) of protein, 3.2 ± 0.85 g (range 1.1-6.1 g) of fat, 6.6 ± 0.34 g of lactose (5.5-8.0 g) and 60 ± 11 kcal (39-94 kcal) of energy per 100 mL. The wide variations in macronutrient content were not influenced by the gestational age of the infant and the lactation day results from 70 of the mothers correlated inversely with the protein content (p < 0.0001; r = -0.42). The MIRIS proved useful, but some adjustments are needed. Variations in macronutrients were high in the breastmilk of women who delivered preterm babies and the protein content decreased with lactation. With adjustments, the MIRIS might provide a helpful tool for individualised fortification. ©2016 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

  5. Negative regulation of parathyroid hormone-related protein expression by steroid hormones.

    PubMed

    Kajitani, Takashi; Tamamori-Adachi, Mimi; Okinaga, Hiroko; Chikamori, Minoru; Iizuka, Masayoshi; Okazaki, Tomoki

    2011-04-15

    Elevated parathyroid hormone-related protein (PTHrP) is responsible for humoral hypercalcemia of malignancy (HHM), which is of clinical significance in treatment of terminal patients with malignancies. Steroid hormones were known to cause suppression of PTHrP expression. However, detailed studies linking multiple steroid hormones to PTHrP expression are lacking. Here we studied PTHrP expression in response to steroid hormones in four cell lines with excessive PTHrP production. Our study established that steroid hormones negatively regulate PTHrP expression. Vitamin D receptor, estrogen receptor α, glucocorticoid receptor, and progesterone receptor, were required for repression of PTHrP expression by the cognate ligands. A notable exception was the androgen receptor, which was dispensable for suppression of PTHrP expression in androgen-treated cells. We propose a pathway(s) involving nuclear receptors to suppress PTHrP expression.

  6. Comparative 2D-DIGE proteomic analysis of bovine mammary epithelial cells during lactation reveals protein signatures for lactation persistency and milk yield.

    PubMed

    Janjanam, Jagadeesh; Singh, Surender; Jena, Manoj K; Varshney, Nishant; Kola, Srujana; Kumar, Sudarshan; Kaushik, Jai K; Grover, Sunita; Dang, Ajay K; Mukesh, Manishi; Prakash, B S; Mohanty, Ashok K

    2014-01-01

    Mammary gland is made up of a branching network of ducts that end with alveoli which surrounds the lumen. These alveolar mammary epithelial cells (MEC) reflect the milk producing ability of farm animals. In this study, we have used 2D-DIGE and mass spectrometry to identify the protein changes in MEC during immediate early, peak and late stages of lactation and also compared differentially expressed proteins in MEC isolated from milk of high and low milk producing cows. We have identified 41 differentially expressed proteins during lactation stages and 22 proteins in high and low milk yielding cows. Bioinformatics analysis showed that a majority of the differentially expressed proteins are associated in metabolic process, catalytic and binding activity. The differentially expressed proteins were mapped to the available biological pathways and networks involved in lactation. The proteins up-regulated during late stage of lactation are associated with NF-κB stress induced signaling pathways and whereas Akt, PI3K and p38/MAPK signaling pathways are associated with high milk production mediated through insulin hormone signaling.

  7. Comparative 2D-DIGE Proteomic Analysis of Bovine Mammary Epithelial Cells during Lactation Reveals Protein Signatures for Lactation Persistency and Milk Yield

    PubMed Central

    Janjanam, Jagadeesh; Singh, Surender; Jena, Manoj K.; Varshney, Nishant; Kola, Srujana; Kumar, Sudarshan; Kaushik, Jai K.; Grover, Sunita; Dang, Ajay K.; Mukesh, Manishi; Prakash, B. S.; Mohanty, Ashok K.

    2014-01-01

    Mammary gland is made up of a branching network of ducts that end with alveoli which surrounds the lumen. These alveolar mammary epithelial cells (MEC) reflect the milk producing ability of farm animals. In this study, we have used 2D-DIGE and mass spectrometry to identify the protein changes in MEC during immediate early, peak and late stages of lactation and also compared differentially expressed proteins in MEC isolated from milk of high and low milk producing cows. We have identified 41 differentially expressed proteins during lactation stages and 22 proteins in high and low milk yielding cows. Bioinformatics analysis showed that a majority of the differentially expressed proteins are associated in metabolic process, catalytic and binding activity. The differentially expressed proteins were mapped to the available biological pathways and networks involved in lactation. The proteins up-regulated during late stage of lactation are associated with NF-κB stress induced signaling pathways and whereas Akt, PI3K and p38/MAPK signaling pathways are associated with high milk production mediated through insulin hormone signaling. PMID:25111801

  8. Continuous PTH and PTHrP infusion causes suppression of bone formation and discordant effects on 1,25(OH)2 vitamin D.

    PubMed

    Horwitz, Mara J; Tedesco, Mary Beth; Sereika, Susan M; Syed, Mushtaq A; Garcia-Ocaña, Adolfo; Bisello, Alessandro; Hollis, Bruce W; Rosen, Clifford J; Wysolmerski, John J; Dann, Pamela; Gundberg, Caren; Stewart, Andrew F

    2005-10-01

    Osteoblast activity and plasma 1,25(OH)2 vitamin D are increased in HPT but suppressed in HHM. To model HPT and HHM, we directly compared multiday continuous infusions of PTH versus PTHrP in humans. Continuous infusion of both PTH and PTHrP results in marked and prolonged suppression of bone formation; renal 1,25(OH)2D synthesis was stimulated effectively by PTH but poorly by PTHrP. PTH and PTH-related protein (PTHrP) cause primary hyperparathyroidism (HPT) and humoral hypercalcemia of malignancy (HHM), respectively. Whereas HHM and HPT resemble one another in many respects, osteoblastic bone formation and plasma 1,25(OH)2 vitamin D are increased in HPT but reduced in HHM. We performed 2- to 4-day continuous infusions of escalating doses of PTH and PTHrP in 61 healthy young adults, comparing the effects on serum calcium and phosphorus, renal calcium and phosphorus handling, 1,25(OH)2 vitamin D, endogenous PTH(1-84) concentrations, and plasma IGF-1 and markers of bone turnover. PTH and PTHrP induced comparable effects on renal calcium and phosphorus handling, and both stimulated IGF-1 and bone resorption similarly. Surprisingly, PTH was consistently more calcemic, reflecting a selectively greater increase in renal 1,25(OH)2 vitamin D production by PTH. Equally surprisingly, continuous infusion of both peptides markedly, continuously, and equivalently suppressed bone formation. PTHrP and PTH produce markedly different effects on 1,25(OH)2 vitamin D homeostasis in humans, leading to different calcemic responses. Moreover, both peptides produce profound suppression of bone formation over multiple days, contrasting with events in HPT, but mimicking HHM. These findings underscore the facts that the mechanisms underlying the anabolic skeletal response to PTH and PTHrP in humans is poorly understood, as are the signal transduction mechanisms that link the renal PTH receptor to 1,25(OH)2 vitamin D synthesis. These studies emphasize that much remains to be learned regarding

  9. Centrosomal proteins and lactate dehydrogenase possess a common epitope in human cell lines.

    PubMed Central

    Gosti, F; Marty, M C; Courvalin, J C; Maunoury, R; Bornens, M

    1987-01-01

    A spontaneously arising rabbit anti-centrosome serum with strong human specificity, used to identify specific antigens in isolated centrosomes, was shown to react with several noncentrosomal proteins including a 36-kDa protein that appeared to be the major cellular antigen. To explore the immunological relationship between noncentrosomal and centrosomal antigens, immunoglobulins were affinity purified using the individual noncentrosomal antigens (from lymphoblastoma KE37 cells) and were tested for their capacity to bind to human centrosomes in situ and to proteins from isolated centrosomes. In this way, the 36-kDa antigen, an abundant cytosolic protein, was shown to share at least one antigenic determinant with high molecular weight centrosomal proteins. This antigen was further identified by mild proteolysis as the glycolytic enzyme lactate dehydrogenase. In all the analyzed human cell lines, the centrosomal staining in situ was correlated with a strong labeling of purified lactate dehydrogenase in immunoblots. Conversely, the absence of centrosomal staining in rodent cells was always correlated with the absence of lactate dehydrogenase labeling. These data suggest an evolutionary relationship between centrosomal proteins and this "housekeeping" enzyme. Images PMID:2434947

  10. Selective protein loss in lactating sows is associated with reduced litter growth and ovarian function.

    PubMed

    Clowes, E J; Aherne, F X; Foxcroft, G R; Baracos, V E

    2003-03-01

    This study was designed to test the degree of protein loss that may be sustained by lactating sows before milk biosynthesis and ovarian function will be impaired. First-parity Camborough x Canabrid sows were allocated to receive isocaloric diets (61 +/- 2.0 MJ of ME/d) and one of three levels of protein intake in lactation: 1) 878 g of CP and 50 g of lysine/d (n = 8), 2) 647 g of CP and 35 g of lysine/d (n = 7), or 3) 491 g of CP and 24 g of lysine/d (n = 10). Every 5 d during a 23-d lactation, sow live weight, backfat depth, and litter weight were recorded, and a preprandial blood sample was collected. Milk samples were collected on d 10 and 20 of lactation. Sows were slaughtered on the day of weaning, and liver and ovarian variables were measured. Lower dietary protein intakes elicited progressively larger live weight losses during lactation (-13, -17, and -28 +/- 2.3 kg; P < 0.001), but similar and minimal backfat losses (-1.3 +/- 0.29 mm). Approximately 7, 9, and 16% of the calculated body protein mass at parturition was mobilized by d 23. Lactation performance did not differ among treatments until d 20, at which time approximately 5, 6, and 12% of the calculated protein mass at parturition had been lost. The milk protein concentration on d 20 of lactation reflected the amount of body protein lost, and was lowest (P < 0.05) in sows that lost the most protein. After d 20, piglet growth rate decreased (P < 0.05) in a manner related to the amount of body protein lost. At weaning, ovarian function was suppressed in sows that had mobilized the most body protein; they had fewer medium-sized follicles (> 4 mm; P < 0.05), their follicles contained less (P < 0.01) follicular fluid, and had lower estradiol (P < 0.05) and IGF-I (P < 0.10) contents. Culture media containing 10% pooled follicular fluid (vol/vol) from high-protein-loss sows were less able to support nuclear and cytoplasmic maturation of oocytes in vitro, evidenced by more oocytes arrested at metaphase I (P

  11. Nitrogen Metabolism in Lactating Goats Fed with Diets Containing Different Protein Sources

    PubMed Central

    Santos, A. B.; Pereira, M. L. A.; Silva, H. G. O.; Pedreira, M. S.; Carvalho, G. G. P.; Ribeiro, L. S. O.; Almeida, P. J. P.; Pereira, T. C. J.; Moreira, J. V.

    2014-01-01

    This study aimed to evaluate urea excretion, nitrogen balance and microbial protein synthesis in lactating goats fed with diets containing different protein sources in the concentrate (soybean meal, cottonseed meal, aerial part of cassava hay and leucaena hay). Four Alpine goats whose mean body weight was 42.6±6.1 kg at the beginning of the experiment, a mean lactation period of 94.0±9.0 days and a production of 1.7±0.4 kg of milk were distributed in a 4×4 Latin square with four periods of 15 days. Diets were formulated to be isonitrogenous, containing 103.0 g/kg of CP, 400 g/kg of Tifton 85 hay and 600 g/kg of concentrate. Diet containing cottonseed meal provided (p<0.05) increased excretion of urea and urea nitrogen in the urine (g/d and mg/kg of BW) when compared with leucaena hay. The diets affected the concentrations of urea nitrogen in plasma (p<0.05) and excretion of urea nitrogen in milk, being that soybean meal and cottonseed meal showed (p<0.05) higher than the average aerial part of the cassava hay. The use of diets with cottonseed meal as protein source in the concentrate in feeding of lactating goats provides greater nitrogen excretion in urine and negative nitrogen balance, while the concentrate with leucaena hay as a source of protein, provides greater ruminal microbial protein synthesis. PMID:25050000

  12. Detection of circulating tumor cells in breast cancer patients using multiplex reverse transcription-polymerase chain reaction and specific primers for MGB, PTHRP and KRT19 correlation with clinicopathological features.

    PubMed

    Skondra, Maria; Gkioka, Eliona; Kostakis, Ioannis D; Pissimissis, Nikolaos; Lembessis, Panagiotis; Pectasides, Dimitrios; Koutsilieris, Michael

    2014-11-01

    The aim of this study was to correlate the clinicopathological features of breast cancer patients with the positive detection of parathyroid hormone-related protein (PTHRP), cytokeratin protein 19 (KRT19) and mammaglobin (MGB) using a multiplex reverse transcription polymerase chain reaction (RT-PCR) assay developed to detect circulating tumor cells (CTCs) in peripheral blood of patients with breast cancer. Peripheral blood samples were collected from 54 breast cancer patients and 20 healthy blood donors. Subsequently, the samples were processed for RNA extraction and analyzed for the expression of PTHRP, KRT19 and MGB using specific primers and multiplex RT-PCR. The positive detection rates in breast cancer patients for PTHRP, KRT19 and MGB were 68.5%, 63% and 22.2% and for healthy donors 10%, 0% and 10%, respectively. The statistical analysis revealed that PTHRP- and KRT19-positive detections correlated with the diagnosis of breast cancer while the combined positive detections of PTHRP-plus-KRT19 correlated with the presence of distant metastasis, especially with bone metastasis. Moreover, positive detections of KRT19 correlated with high proliferation rate of breast cancer tumors. MGB-positive detections did not add any diagnostic advantage in such analysis. Multiplex-PCR based detection of CTCs using PTHRP and KRT19 primers can provide useful information for the disease. Copyright© 2014 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

  13. Monotreme lactation protein is highly expressed in monotreme milk and provides antimicrobial protection.

    PubMed

    Enjapoori, Ashwantha Kumar; Grant, Tom R; Nicol, Stewart C; Lefèvre, Christophe M; Nicholas, Kevin R; Sharp, Julie A

    2014-09-22

    Monotremes (platypus and echidna) are the descendants of the oldest ancestor of all extant mammals distinguished from other mammals by mode of reproduction. Monotremes lay eggs following a short gestation period and after an even briefer incubation period, altricial hatchlings are nourished over a long lactation period with milk secreted by nipple-less mammary patches located on the female's abdomen. Milk is the sole source of nutrition and immune protection for the developing young until weaning. Using transcriptome and mass spectrometry analysis of milk cells and milk proteins, respectively, a novel Monotreme Lactation Protein (MLP) was identified as a major secreted protein in milk. We show that platypus and short-beaked echidna MLP genes show significant homology and are unique to monotremes. The MLP transcript was shown to be expressed in a variety of tissues; however, highest expression was observed in milk cells and was expressed constitutively from early to late lactation. Analysis of recombinant MLP showed that it is an N-linked glycosylated protein and biophysical studies predicted that MLP is an amphipathic, α-helical protein, a typical feature of antimicrobial proteins. Functional analysis revealed MLP antibacterial activity against both opportunistic pathogenic Staphylococcus aureus and commensal Enterococcus faecalis bacteria but showed no effect on Escherichia coli, Pseudomonas aeruginosa, Staphylococcus epidermidis, and Salmonella enterica. Our data suggest that MLP is an evolutionarily ancient component of milk-mediated innate immunity absent in other mammals. We propose that MLP evolved specifically in the monotreme lineage supporting the evolution of lactation in these species to provide bacterial protection, at a time when mammals lacked nipples. © The Author(s) 2014. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  14. Monotreme Lactation Protein Is Highly Expressed in Monotreme Milk and Provides Antimicrobial Protection

    PubMed Central

    Enjapoori, Ashwantha Kumar; Grant, Tom R.; Nicol, Stewart C.; Lefèvre, Christophe M.; Nicholas, Kevin R.; Sharp, Julie A.

    2014-01-01

    Monotremes (platypus and echidna) are the descendants of the oldest ancestor of all extant mammals distinguished from other mammals by mode of reproduction. Monotremes lay eggs following a short gestation period and after an even briefer incubation period, altricial hatchlings are nourished over a long lactation period with milk secreted by nipple-less mammary patches located on the female’s abdomen. Milk is the sole source of nutrition and immune protection for the developing young until weaning. Using transcriptome and mass spectrometry analysis of milk cells and milk proteins, respectively, a novel Monotreme Lactation Protein (MLP) was identified as a major secreted protein in milk. We show that platypus and short-beaked echidna MLP genes show significant homology and are unique to monotremes. The MLP transcript was shown to be expressed in a variety of tissues; however, highest expression was observed in milk cells and was expressed constitutively from early to late lactation. Analysis of recombinant MLP showed that it is an N-linked glycosylated protein and biophysical studies predicted that MLP is an amphipathic, α-helical protein, a typical feature of antimicrobial proteins. Functional analysis revealed MLP antibacterial activity against both opportunistic pathogenic Staphylococcus aureus and commensal Enterococcus faecalis bacteria but showed no effect on Escherichia coli, Pseudomonas aeruginosa, Staphylococcus epidermidis, and Salmonella enterica. Our data suggest that MLP is an evolutionarily ancient component of milk-mediated innate immunity absent in other mammals. We propose that MLP evolved specifically in the monotreme lineage supporting the evolution of lactation in these species to provide bacterial protection, at a time when mammals lacked nipples. PMID:25245409

  15. Secretion of whey acidic protein and cystatin is down regulated at mid-lactation in the red kangaroo (Macropus rufus)

    USGS Publications Warehouse

    Nicholas, K.R.; Fisher, J.A.; Muths, E.; Trott, J.; Janssens, P.A.; Reich, C.; Shaw, D.C.

    2001-01-01

    Milk collected from the red kangaroo (Macropus rufus) between day 100 and 260 of lactation showed major changes in milk composition at around day 200 of lactation, the time at which the pouch young begins to temporarily exit the pouch and eat herbage. The carbohydrate content of milk declined abruptly at this time and although there was only a small increase in total protein content, SDS PAGE analysis of milk revealed asynchrony in the secretory pattern of individual proteins. The levels of ??-lactalbumin, ??-lactoglobulin, serum albumin and transferrin remain unchanged during lactation. In contrast, the protease inhibitor cystatin, and the putative protease inhibitor whey acidic protein (WAP) first appeared in milk at elevated concentrations after approximately 150 days of lactation and then ceased to be secreted at approximately 200 days. In addition, a major whey protein, late lactation protein, was first detected in milk around the time whey acidic protein and cystatin cease to be secreted and was present at least until day 260 of lactation. The co-ordinated, but asynchronous secretion of putative protease inhibitors in milk may have several roles during lactation including tissue remodelling in the mammary gland and protecting specific proteins in milk required for physiological development of the dependent young. ?? 2001 Elsevier Science Inc.

  16. Secretion of whey acidic protein and cystatin is down regulated at mid-lactation in the red kangaroo (Macropus rufus)

    USGS Publications Warehouse

    Nicholas, K.R.; Fisher, J.A.; Muths, E.; Trott, J.; Janssens, P.A.; Reich, C.; Shaw, D.C.

    2001-01-01

    Milk collected from the red kangaroo (Macropus rufus) between day 100 and 260 of lactation showed major changes in milk composition at around day 200 of lactation, the time at which the pouch young begins to temporarily exit the pouch and eat herbage. The carbohydrate content of milk declined abruptly at this time and although there was only a small increase in total protein content, SDS PAGE analysis of milk revealed asynchrony in the secretory pattern of individual proteins. The levels of α-lactalbumin, β-lactoglobulin, serum albumin and transferrin remain unchanged during lactation. In contrast, the protease inhibitor cystatin, and the putative protease inhibitor whey acidic protein (WAP) first appeared in milk at elevated concentrations after approximately 150 days of lactation and then ceased to be secreted at approximately 200 days. In addition, a major whey protein, late lactation protein, was first detected in milk around the time whey acidic protein and cystatin cease to be secreted and was present at least until day 260 of lactation. The co-ordinated, but asynchronous secretion of putative protease inhibitors in milk may have several roles during lactation including tissue remodelling in the mammary gland and protecting specific proteins in milk required for physiological development of the dependent young.

  17. Maternal protein reserves and their influence on lactational performance in rats. 2. Effects of dietary protein restriction during gestation and lactation on tissue protein metabolism and Na+, K(+)-ATPase (EC 3.6.1.3) activity.

    PubMed

    Pine, A P; Jessop, N S; Allan, G F; Oldham, J D

    1994-08-01

    Changes in tissue protein synthesis and an associated membrane transport system in rats were investigated during lactation and under conditions of dietary protein restriction. Following mating, female Sprague-Dawley rats (second parity) were caged individually and offered a high-protein diet (H; 215 g crude protein (N x 6.25; CP)/kg dry matter (DM)) ad lib. until day 12 of gestation. Subsequently half continued to receive diet H, whilst the remainder were offered a low-protein diet (L; 65 g CP/kg DM) until parturition. On day 1 of lactation females were then allocated to either diet H or another low-protein diet (L2; 90 g CP/kg DM) which were offered ad lib. until day 13 of lactation, giving four lactation groups HH, LH, HL2 and LL2. On days 1 and 13 of lactation groups of females were used in the estimation of tissue protein synthesis (flooding dose of [3H] phenylalanine) and Na+, K(+)-ATPase (EC 3.6.1.3) activity (polarographically) in skeletal muscle, mammary gland, liver and duodenal mucosa. By day 1 of lactation diet L had reduced fractional and absolute synthesis rates (FSR and ASR) of muscle protein (P < 0.05) and the O2 consumption associated with Na+, K(+)-ATPase, although not significantly (P < 0.10). Rates of protein synthesis in the other tissues studied were not affected on day 1 of lactation by the gestation dietary treatment. By day 13 of lactation the feeding of diet L2 had reduced muscle FSR and ASR of group HL2 to rates that were lower than those on day 1 (P < 0.05), comparable to those of group LL2 and lower than those of groups HH and LH (P < 0.05). Diet H had allowed group LH to increase their muscle protein synthesis compared with that on day 1 (P < 0.05). Muscle Na+, K(+)-ATPase activity on day 13 of lactation was also lower in groups offered diet L2 (P < 0.05). Mammary protein synthesis was increased during lactation with the feeding of diet H (P < 0.05), which was prevented by diet L2 such that rates of groups HL2 and LL2 were lower than

  18. Effect of varying protein and energy densities in complete rations fed to cows in first lactation.

    PubMed

    Macleod, G K; Grieve, D G; McMillan, I; Smith, G C

    1984-07-01

    Sixty-three Holstein cows in first lactation were allotted randomly at 28 days postpartum to receive one of nine rations in a 3 X 3 factorial design. Rations had crude protein 12, 15, or 18% in each of three energy densities from ratios of forage:concentrate of 75:25, 55:45, and 35:65. Corn silage and hay crop silage were combined in a ratio of 2:1 (dry basis) in each case, and diets were offered for ad libitum intake as complete feeds for an 11-wk experimental period plus 2 wk for a digestion trial. In regression analysis as energy density increased, feed intake, milk yield, milk protein, and lactose yields increased linearly, urea nitrogen in blood plasma decreased linearly, and milk fat percent and yield decreased curvilinearly. As dietary protein increased, feed intake, fat-corrected milk, milk fat yield, and plasma urea nitrogen increased linearly, and yields of milk, solids-corrected milk, milk protein, and lactose increased curvilinearly. Actual milk yield adjusted for dry matter intake increased with greater energy density but not with higher protein concentration. Increases of energy and protein each resulted in linear increases of digestibility of dry matter, crude protein, and energy of diets. There were no interactions of energy X protein for these measurements. Cows in first lactation will increase milk production in response to increasing protein over a range of energy densities when feed intake is increased.

  19. C-reactive protein by pregnancy and lactational status among Filipino young adult women

    PubMed Central

    Kuzawa, Christopher W.; Adair, Linda S.; Borja, Judith; McDade, Thomas W

    2013-01-01

    Objectives Pregnancy and lactation involve adaptations in immune regulation, but little is known about cross-cultural variation in inflammatory changes during pregnancy or lactation. Here we report concentrations of C-reactive protein (CRP) in a large cross-sectional sample of healthy Filipino women who vary in parity, gestational and lactational status, and who come from a population previously described as having low CRP. Methods Fasting plasma CRP was measured among female participants (ages 20.8-22.4 years) in the Cebu Longitudinal Health and Nutrition Survey (n=822). Results Median CRP was 0.2 mg/L in nulliparous women and peaked at 2.0 mg/L in women in their 3rd trimester of pregnancy. Parous but post-partum women had higher CRP compared to nulliparous women, which was largely explained by body composition differences as reflected in waist circumference and skinfold measures. Among post-partum women with infants, CRP was similar in women who were currently breastfeeding compared to those who were not. Conclusions At Cebu, women late in gestation have 10-fold higher C-reactive protein compared to nulliparous women, with no evidence that lactation is inflammatory. These population-based findings are similar with findings from prior clinic-based studies and are consistent with the maternal immunological adaptations initiated during pregnancy. The tendency of human females to spend more time than females of other great apes in gestation rather than lactation suggests that the human life history strategy involved increased time spent by reproductively aged females in a pro-inflammatory state. PMID:23180717

  20. Parathyroid Hormone-Related Protein Interacts With the Transforming Growth Factor-β/Bone Morphogenetic Protein-2/Gremlin Signaling Pathway to Regulate Proinflammatory and Profibrotic Mediators in Pancreatic Acinar and Stellate Cells.

    PubMed

    Bhatia, Vandanajay; Cao, Yanna; Ko, Tien C; Falzon, Miriam

    2016-01-01

    Transforming growth factor β (TGF-β) regulates immune and fibrotic responses of chronic pancreatitis. The bone morphogenetic protein 2 (BMP-2) antagonist gremlin is regulated by TGF-β. Parathyroid hormone-related protein (PTHrP) levels are elevated in chronic pancreatitis. Here, we investigated the cross-talk between TGF-β/BMP-2/gremlin and PTHrP signaling. Reverse transcription/real-time polymerase chain reaction, chromatin immunoprecipitation, Western blotting, and transient transfection were used to investigate PTHrP regulation by TGF-β and BMP-2 and gremlin regulation by PTHrP. The PTHrP antagonist PTHrP (7-34) and acinar cells with conditional Pthrp gene deletion (PTHrP) were used to assess PTHrP's role in the proinflammatory and profibrotic effects of TGF-β and gremlin. Transforming growth factor β increased PTHrP levels in acinar cells and pancreatic stellate cells (PSCs) through a Smad3-dependent pathway. Transforming growth factor β's effects on levels of IL-6 and intercellular adhesion molecule 1 (ICAM-1) (acinar cells) and procollagen I and fibronectin (PSCs) were inhibited by PTHrP (7-34). PTHrP suppressed TGF-β's effects on IL-6 and ICAM-1. Parathyroid hormone-related hormone increased gremlin in acinar cells, and inhibiting gremlin action suppressed TGF-β's and PTHrP's effects on IL-6 and ICAM-1. Transforming growth factor β-mediated gremlin up-regulation was suppressed in PTHrP cells. Bone morphogenetic protein 2 suppressed PTHrP levels in PSCs. Parathyroid hormone-related hormone functions as a novel mediator of the proinflammatory and profibrotic effects of TGF-β. Transforming growth factor β and BMP-2 regulate PTHrP expression, and PTHrP regulates gremlin levels.

  1. Protein quality and quantity and insulin control of mammary gland glucose utilization during lactation

    SciTech Connect

    Masor, M.L.

    1987-01-01

    Virgin Sprague-Dawley rats were bred, and fed laboratory stock (STOCK), 13% casein plus methionine, 13% wheat gluten, or 5% casein plus methionine through gestation and 4 days of lactation. Diets were switched at parturition to determine the effects of dietary protein quality and quantity fed during gestation and/or lactation on insulin stimulation of mammary glucose utilization. On day 20 of gestation (20G) and day 4 of lactation (4L) the right inguinal-abdominal mammary glands were removed, and acini and tissue slices were incubated in Krebs buffer with or without insulin containing (U-/sup 14/C)-glucose and 5mM glucose for 1 hour at 37/degrees/C. Glucose incorporation into CO/sub 2/, lipid and lactose was determined. Glucose incorporation into CO/sub 2/ and lipid, but not lactose was stimulated by insulin in mammary slices. Diet effects on glucose utilization in acini were confirmed in slices for basal and insulin stimulated levels. Treatment affected the absolute increase of insulin stimulation. Regression analysis significantly correlated pup weight gain with total glucose utilization. Poor dietary protein quality and quantity fed during gestation impaired both overall response of mammary glucose utilization to insulin stimulation, and mammary development during pregnancy. Improving protein value at parturition did not overcome those deficits by 4L.

  2. Hormonal regulation of platypus Beta-lactoglobulin and monotreme lactation protein genes.

    PubMed

    Enjapoori, Ashwantha Kumar; Lefèvre, Christophe M; Nicholas, Kevin R; Sharp, Julie A

    2017-02-01

    Endocrine regulation of milk protein gene expression in marsupials and eutherians is well studied. However, the evolution of this complex regulation that began with monotremes is unknown. Monotremes represent the oldest lineage of extant mammals and the endocrine regulation of lactation in these mammals has not been investigated. Here we characterised the proximal promoter and hormonal regulation of two platypus milk protein genes, Beta-lactoglobulin (BLG), a whey protein and monotreme lactation protein (MLP), a monotreme specific milk protein, using in vitro reporter assays and a bovine mammary epithelial cell line (BME-UV1). Insulin and dexamethasone alone provided partial induction of MLP, while the combination of insulin, dexamethasone and prolactin was required for maximal induction. Partial induction of BLG was achieved by insulin, dexamethasone and prolactin alone, with maximal induction using all three hormones. Platypus MLP and BLG core promoter regions comprised transcription factor binding sites (e.g. STAT5, NF-1 and C/EBPα) that were conserved in marsupial and eutherian lineages that regulate caseins and whey protein gene expression. Our analysis suggests that insulin, dexamethasone and/or prolactin alone can regulate the platypus MLP and BLG gene expression, unlike those of therian lineage. The induction of platypus milk protein genes by lactogenic hormones suggests they originated before the divergence of marsupial and eutherians.

  3. Genes regulating lipid and protein metabolism are highly expressed in mammary gland of lactating dairy goats.

    PubMed

    Shi, Hengbo; Zhu, Jiangjiang; Luo, Jun; Cao, Wenting; Shi, Huaiping; Yao, Dawei; Li, Jun; Sun, Yuting; Xu, Huifen; Yu, Kang; Loor, Juan J

    2015-05-01

    Dairy goats serve as an important source of milk and also fulfill agricultural and economic roles in developing countries. Understanding the genetic background of goat mammary gland is important for research on the regulatory mechanisms controlling tissue function and the synthesis of milk components. We collected tissue at four different stages of goat mammary gland development and generated approximately 25 GB of data from Illumina de novo RNA sequencing. The combined reads were assembled into 51,361 unigenes, and approximately 60.07 % of the unigenes had homology to other proteins in the NCBI non-redundant protein database (NR). Functional classification through eukaryotic Ortholog Groups of Protein (KOG), gene ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed that the unigenes from goat mammary glands are involved in a wide range of biological processes and metabolic pathways, including lipid metabolism and lactose metabolism. The results of qPCR revealed that genes encoding FABP3, FASN, SCD, PLIN2, whey proteins (LALBA and BLG), and caseins (CSN1S1, CSN1S2, CSN2 and CSN3) at 100 and 310 days postpartum increased significantly compared with the non-lactating period. In addition to their role in lipid and protein synthesis, the higher expression at 310 days postpartum could contribute to mammary cell turnover during pregnancy. In conclusion, this is the first study to characterize the complete transcriptome of goat mammary glands and constitutes a comprehensive genomic resource available for further studies of ruminant lactation.

  4. Effect of osmolytes on protein dynamics in the lactate dehydrogenase-catalyzed reaction.

    PubMed

    Zhadin, Nickolay; Callender, Robert

    2011-03-15

    Laser-induced temperature jump relaxation spectroscopy was used to probe the effect of osmolytes on the microscopic rate constants of the lactate dehydrogenase-catalyzed reaction. NADH fluorescence and absorption relaxation kinetics were measured for the lactate dehydrogenase (LDH) reaction system in the presence of varying amounts of trimethylamine N-oxide (TMAO), a protein-stabilizing osmolyte, or urea, a protein-destabilizing osmolyte. Trimethylamine N-oxide (TMAO) at a concentration of 1 M strongly increases the rate of hydride transfer, nearly nullifies its activation energy, and also slightly increases the enthalpy of hydride transfer. In 1 M urea, the hydride transfer enthalpy is almost nullified, but the activation energy of the step is not affected significantly. TMAO increases the preference of the closed conformation of the active site loop in the LDH·NAD(+)·lactate complex; urea decreases it. The loop opening rate in the LDH·NADH·pyruvate complex changes its temperature dependence to inverse Arrhenius with TMAO. In this complex, urea accelerates the loop motion, without changing the loop opening enthalpy. A strong, non-Arrhenius decrease in the pyruvate binding rate in the presence of TMAO offers a decrease in the fraction of the open loop, pyruvate binding competent form at higher temperatures. The pyruvate off rate is not affected by urea but decreases with TMAO. Thus, the osmolytes strongly affect the rates and thermodynamics of specific events along the LDH-catalyzed reaction: binding of substrates, loop closure, and the chemical event. Qualitatively, these results can be understood as an osmolyte-induced change in the energy landscape of the protein complexes, shifting the conformational nature of functional substates within the protein ensemble.

  5. Exogenous PTHrP Repairs the Damaged Fracture Healing of PTHrP+/− Mice and Accelerates Fracture Healing of Wild Mice

    PubMed Central

    Wang, Yinhe; Fang, Xin; Wang, Chun; Ding, Congzhu; Lin, Hua; Liu, Anlong; Wang, Lei; Cao, Yang

    2017-01-01

    Bone fracture healing is a complicated physiological regenerative process initiated in response to injury and is similar to bone development. To demonstrate whether an exogenous supply of parathyroid hormone–related protein (PTHrP) helps in bone fracture healing, closed mid-diaphyseal femur fractures were created and stabilized with intramedullary pins in eight-week-old wild-type (WT) PTHrP+/+ and PTHrP+/− mice. After administering PTHrP for two weeks, callus tissue properties were analyzed at one, two, and four weeks post-fracture (PF) by various methods. Bone formation–related genes and protein expression levels were evaluated by real-time reverse transcriptase–polymerase chain reaction and Western blots. At two weeks PF, mineral density of callus, bony callus areas, mRNA levels of alkaline phosphatase (ALP), type I collagen, Runt-related transcription factor 2 (Runx-2), and protein levels of Runx-2 and insulin-like growth factor-1 decreased in PTHrP+/− mice compared with WT mice. At four weeks PF, total collagen-positive bony callus areas, osteoblast number, ALP-positive areas, and type I collagen-positive areas all decreased in PTHrP+/− mice. At both two and four weeks PF, tartrate-resistant acid phosphatase–positive osteoclast number and surface decreased a little in PTHrP+/− mice. The study indicates that exogenous PTHrP provided by subcutaneous injection could redress impaired bone fracture healing, leading to mutation of activated PTHrP by influencing callus areas, endochondral bone formation, osteoblastic bone formation, and bone turnover. PMID:28178186

  6. The effect of protein supplementation on lactate accumulation during submaximal and maximal exercise.

    PubMed

    Vukovich, M D; Sharp, R L; King, D S; Kershishnik, K

    1992-12-01

    Eleven subjects performed a graded exercise test after 1 week of protein supplementation (PRO) or glucose polymer placebo (CON), randomly assigned in a double blind fashion. The exercise consisted of 3-min graded exercise bouts separated by 10 min of active recovery at zero pedal resistance. Subjects then performed a 30-sec Wingate test (WIN) to assess performance during supramaximal exercise. Blood samples were obtained in the last 15 sec of each exercise and recovery period. PRO resulted in a decrease in blood lactate following 120% VO2max and WIN, an increase in blood alanine at all time points, and lower postexercise muscle lactate and glycogen. Resting muscle GPT activity was 47% higher during the PRO trial. Mean power output during the WIN did not differ between PRO and CON. The WIN fatigue index was not significantly different between PRO and CON. The increased alanine may reflect increased transamination of pyruvate, thereby reducing the accumulation of lactate, which in turn had a marginal effect on performance during supramaximal exercise.

  7. Engraftment and bone mass are enhanced by PTHrP 1-34 in ectopically transplanted vertebrae (vossicle model) and can be non-invasively monitored with bioluminescence and fluorescence imaging.

    PubMed

    Hildreth, Blake Eason; Williams, Michelle M; Dembek, Katarzyna A; Hernon, Krista M; Rosol, Thomas J; Toribio, Ramiro E

    2015-12-01

    Evidence exists that parathyroid hormone-related protein (PTHrP) 1-34 may be more anabolic in bone than parathyroid hormone 1-34. While optical imaging is growing in popularity, scant information exists on the relationships between traditional bone imaging and histology and bioluminescence (BLI) and fluorescence (FLI) imaging. We aimed to evaluate the effects of PTHrP 1-34 on bone mass and determine if relationships existed between radiographic and histologic findings in bone and BLI and FLI indices. Vertebrae (vossicles) from mice coexpressing luciferase and green fluorescent protein were implanted subcutaneously into allogenic nude mice. Transplant recipients were treated daily with saline or PTHrP 1-34 for 4 weeks. BLI, FLI, radiography, histology, and µCT of the vossicles were performed over time. PTHrP 1-34 increased bioluminescence the most after 2 weeks, fluorescence at all time points, and decreased the time to peak bioluminescence at 4 weeks (P ≤ 0.027), the latter of which suggesting enhanced engraftment. PTHrP 1-34 maximized vertebral body volume at 4 weeks (P < 0.0001). The total amount of bone observed histologically increased in both groups at 2 and 4 weeks (P ≤ 0.002); however, PTHrP 1-34 exceeded time-matched controls (P ≤ 0.044). A positive linear relationship existed between the percentage of trabecular bone and (1) total bioluminescence (r = 0.595; P = 0.019); (2) total fluorescence (r = 0.474; P = 0.074); and (3) max fluorescence (r = 0.587; P = 0.021). In conclusion, PTHrP 1-34 enhances engraftment and bone mass, which can be monitored non-invasively by BLI and FLI.

  8. Engraftment and bone mass are enhanced by PTHrP 1-34 in ectopically transplanted vertebrae (vossicle model) and can be non-invasively monitored with bioluminescence and fluorescence imaging

    PubMed Central

    Hildreth, Blake Eason; Williams, Michelle C.; Dembek, Katarzyna A.; Hernon, Krista M.; Rosol, Thomas J.; Toribio, Ramiro E.

    2015-01-01

    Evidence exists that parathyroid hormone-related protein (PTHrP) 1-34 may be more anabolic in bone than parathyroid hormone (PTH) 1-34. While optical imaging is growing in popularity, scant information exists on the relationships between traditional bone imaging and histology and bioluminescence (BLI) and fluorescence (FLI) imaging. We aimed to evaluate the effects of PTHrP 1-34 on bone mass and determine if relationships existed between radiographic and histologic findings in bone and BLI and FLI indices. Vertebrae (vossicles) from mice coexpressing luciferase and green fluorescent protein were implanted subcutaneously into allogenic nude mice. Transplant recipients were treated daily with saline or PTHrP 1-34 for 4 weeks. BLI, FLI, radiography, histology, and μCT of the vossicles were performed over time. PTHrP 1-34 increased bioluminescence the most after 2 weeks, fluorescence at all time points, and decreased the time to peak bioluminescence at 4 weeks (P ≤ 0.027), the latter of which suggesting enhanced engraftment. PTHrP 1-34 maximized vertebral body volume at 4 weeks (P < 0.0001). The total amount of bone observed histologically increased in both groups at 2 and 4 weeks (P ≤ 0.002); however, PTHrP 1-34 exceeded time-matched controls (P ≤ 0.044). A positive linear relationship existed between the percentage of trabecular bone and 1) total bioluminescence (r = 0.595; P = 0.019); 2) total fluorescence (r = 0.474; P = 0.074); and 3) max fluorescence (r = 0.587; P = 0.021). In conclusion, PTHrP 1-34 enhances engraftment and bone mass, which can be monitored non-invasively by BLI and FLI. PMID:26271486

  9. Involvement of ERK1/2, p38 MAPK, and PI3K/Akt signaling pathways in the regulation of cell cycle progression by PTHrP in colon adenocarcinoma cells.

    PubMed

    Calvo, Natalia; Martín, María Julia; de Boland, Ana Russo; Gentili, Claudia

    2014-08-01

    Parathyroid hormone-related peptide (PTHrP) is distributed in most fetal and adult tissues, and its expression correlates with the severity of colon carcinoma. Recently we obtained evidence that in Caco-2 cells, a cell line from human colorectal adenocarcinoma, exogenous PTHrP increases the number of live cells, via ERK1/2, p38 MAPK, and PI3-kinase and induces the expression of cyclin D1, a cell cycle regulatory protein. In this study, we further investigated the role of PTHrP in the regulation of the cell cycle progression in these intestinal cells. Flow cytometry analysis revealed that PTHrP treatment diminishes the number of cells in the G0/G1 phase and increases the number in both S and G2/M phases. The hormone increases the expression of CDK6 and diminishes the amount of negative cell cycle regulators p27Kip1, p15INK4B, and p53. However, PTHrP does not modify the expression of cyclin D3, CDK4, and p16INK4A. In addition, inhibitors of ERK1/2 (PD98059), p38 MAPK (SB203580), and PI3Kinase (LY294002) reversed PTHrP response in Caco-2 cells. Taken together, our results suggest that PTHrP positively modulates cell cycle progression and changes the expression of proteins involved in cell cycle regulation via ERK1/2, p38 MAPK, and PI3K signaling pathways in Caco-2 cells.

  10. Calcium handling in Sparus auratus: effects of water and dietary calcium levels on mineral composition, cortisol and PTHrP levels.

    PubMed

    Abbink, Wout; Bevelander, Gideon S; Rotllant, Josep; Canario, Adelino V M; Flik, Gert

    2004-11-01

    Juvenile gilthead sea bream (Sparus auratus L.; 10-40 g body mass) were acclimatized in the laboratory to full strength (34 per thousand) or dilute (2.5 per thousand) seawater and fed normal, calcium-sufficient or calcium-deficient diet for nine weeks. Mean growth rate, whole-body calcium and phosphorus content and accumulation rates were determined, as well as plasma levels of ionic and total calcium, cortisol and parathyroid hormone related protein (PTHrP; a hypercalcemic hormone in fish). When confronted with limited calcium access (low salinity and calcium-deficient diet), sea bream show growth arrest. Both plasma cortisol and PTHrP increase when calcium is limited in water or diet, and a positive relationship was found between plasma PTHrP and plasma ionic calcium (R(2)=0.29, N=18, P<0.05). Furthermore, a strong correlation was found between net calcium and phosphorus accumulation (R(2)=0.92, N=16, P<0.01) and between body mass and whole-body calcium (R(2)=0.84, N=25, P<0.01) and phosphorus (R(2)=0.88, N=24, P<0.01) content. Phosphorus accumulation is strongly calcium dependent, as phosphorus accumulation decreases in parallel to calcium accumulation when the diet is calcium deficient but phosphorus sufficient. We conclude that PTHrP and cortisol are involved in the regulation of the hydromineral balance of these fish, with growth-related calcium accumulation as an important target.

  11. Cellular prion protein directly interacts with and enhances lactate dehydrogenase expression under hypoxic conditions.

    PubMed

    Ramljak, Sanja; Schmitz, Matthias; Zafar, Saima; Wrede, Arne; Schenkel, Sara; Asif, Abdul R; Carimalo, Julie; Doeppner, Thorsten R; Schulz-Schaeffer, Walter J; Weise, Jens; Zerr, Inga

    2015-09-01

    Although a physiological function of the cellular prion protein (PrP(c)) is still not fully clarified, a PrP(c)-mediated neuroprotection against hypoxic/ischemic insult is intriguing. After ischemic stroke prion protein knockout mice (Prnp(0/0)) display significantly greater lesions as compared to wild-type (WT) mice. Earlier reports suggested an interaction between the glycolytic enzyme lactate dehydrogenase (LDH) and PrP(c). Since hypoxic environment enhances LDH expression levels and compels neurons to rely on lactate as an additional oxidative substrate for energy metabolism, we examined possible differences in LDH protein expression in WT and Prnp(0/0) knockout models under normoxic/hypoxic conditions in vitro and in vivo, as well as in a HEK293 cell line. While no differences are observed under normoxic conditions, LDH expression is markedly increased after 60-min and 90-min of hypoxia in WT vs. Prnp(0/0) primary cortical neurons with concurrent less hypoxia-induced damage in the former group. Likewise, cerebral ischemia significantly increases LDH levels in WT vs. Prnp(0/0) mice with accompanying smaller lesions in the WT group. HEK293 cells overexpressing PrP(c) show significantly higher LDH expression/activity following 90-min of hypoxia as compared to control cells. Moreover, a cytoplasmic co-localization of LDH and PrP(c) was recorded under both normoxic and hypoxic conditions. Interestingly, an expression of monocarboxylate transporter 1, responsible for cellular lactate uptake, increases with PrP(c)-overexpression under normoxic conditions. Our data suggest LDH as a direct PrP(c) interactor with possible physiological relevance under low oxygen conditions.

  12. Mechanisms of vasodilation to PTH 1-84, PTH 1-34, and PTHrP 1-34 in rat bone resistance arteries.

    PubMed

    Benson, T; Menezes, T; Campbell, J; Bice, A; Hood, B; Prisby, R

    2016-05-01

    Parathyroid hormone (PTH) augments bone metabolism and bone mass when given intermittently. Enhanced blood flow is requisite to support high tissue metabolism. The bone arteries are responsive to all three PTH analogs, which may serve to augment skeletal blood flow during intermittent PTH administration. PTH augments bone metabolism. Yet, mechanisms by which PTH regulates bone blood vessels are unknown. We deciphered (1) endothelium-dependent and endothelium-independent vasodilation to PTH 1-84, PTH 1-34, and PTHrP 1-34, (2) the signaling pathways (i.e., endothelial nitric oxide synthase [eNOS], cyclooxygenase [COX], protein kinase C [PKC], and protein kinase A [PKA]), and (3) receptor activation. Femoral principal nutrient arteries (PNAs) were given cumulative doses (10(-13)-10(-8) M) of PTH 1-84, PTH 1-34, and PTHrP 1-34 with and without signaling pathway blockade. Vasodilation was also determined following endothelial cell removal (i.e., denudation), PTH 1 receptor (PTH1R) inhibition and to sodium nitroprusside (SNP; a nitric oxide [NO] donor). Vasodilation was lowest to PTH 1-34, and maximal dilation was highest to PTHrP 1-34. Inhibition of eNOS reduced vasodilation to PTH 1-84 (-80 %), PTH 1-34 (-66 %), and PTHrP 1-34 (-48 %), evidencing the contribution of NO. Vasodilation following denudation was eliminated (PTH 1-84 and PTHrP 1-34) and impaired (PTH 1-34, 17 % of maximum), highlighting the importance of endothelial cells for PTH signaling. Denuded and intact PNAs responded similarly to SNP. Both PKA and PKC inhibition diminished vasodilation in all three analogs to varying degrees. PTH1R blockade reduced vasodilation to 1, 12, and 12 % to PTH 1-84, PTH 1-34, and PTHrP 1-34, respectively. Vasodilation of femoral PNAs to the PTH analogs occurred via activation of the endothelial cell PTH1R for NO-mediated events. PTH 1-84 and PTHrP 1-34 primarily stimulated PKA signaling, and PTH 1-34 equally stimulated PKA and PKC signaling.

  13. Induction of Thermal and Mechanical Hypersensitivity by Parathyroid Hormone-related Peptide (PTHrP) Through Upregulation of TRPV1 Function and Trafficking

    PubMed Central

    Mickle, Aaron D.; Shepherd, Andrew J.; Loo, Lipin; Mohapatra, Durga P.

    2016-01-01

    The neurobiological mechanisms underlying chronic pain associated with cancers are not well understood. It has been hypothesized that factors specifically elevated in the tumor microenvironment sensitize adjacent nociceptive afferents. We show that parathyroid hormone-related peptide (PTHrP), which is found at elevated levels in the tumor microenvironment of advanced breast and prostate cancers, is a critical modulator of sensory neurons. Intraplantar injection of PTHrP led to the development of thermal and mechanical hypersensitivity in both male and female mice, which were absent in mice lacking functional transient receptor potential vanilloid-1 (TRPV1). The PTHrP treatment of cultured mouse sensory neurons enhanced action potential firing, and increased TRPV1 activation, which was dependent on protein kinase C (PKC) activity. Parathyroid hormone-related peptide induced robust potentiation of TRPV1 activation and enhancement of neuronal firing at mild acidic pH that is relevant to acidic tumor microenvironment. We also observed an increase in plasma membrane TRPV1 protein levels after exposure to PTHrP, leading to upregulation in the proportion of TRPV1-responsive neurons, which was dependent on the activity of PKC and Src kinases. Furthermore, co-injection of PKC or Src inhibitors attenuated PTHrP-induced thermal but not mechanical hypersensitivity. Altogether, our results suggest that PTHrP and mild acidic conditions could induce constitutive pathological activation of sensory neurons through upregulation of TRPV1 function and trafficking, which could serve as a mechanism for peripheral sensitization of nociceptive afferents in the tumor microenvironment. PMID:25970319

  14. Lactation curves of Valle del Belice dairy ewes for yields of milk, fat, and protein estimated with test day models.

    PubMed

    Cappio-Borlino, A; Portolano, B; Todaro, M; Macciotta, N P; Giaccone, P; Pulina, G

    1997-11-01

    Test day models were used to estimate the lactation curves for Valle del Belice ewes and to study the main environmental effects on milk yield and on percentage of fat and protein. Environmental effects were treated as fixed. A random effect was associated with each lactation to evaluate the mean correlation among all test day records of an individual ewe. Lactation curves were constructed by adding solutions for classes of either days in milk nested within parity or days in milk nested within season of lambing to appropriate general means. Parity primarily affected the lactation curve for milk yield, which was lower and flatter for first lambing ewes; effects on fat and protein were smaller. Season of lambing affected all traits. Seasonal productivity had the greatest effect on milk composition, resulting in an imbalance between fat and protein percentages. Flock and feed supplementation affected only the lactation curve for milk yield. The lactation curve of Valle del Belice ewes stood at a relatively high level. However, the presence of notable, perturbative effects (environmental and random variation) on milk yield and composition suggests that management is unable to meet the requirements of ewes consistently.

  15. Stage of lactation and corresponding diets affect in situ protein degradation by dairy cows.

    PubMed

    Schadt, I; Mertens, D R; Van Soest, P J; Azzaro, G; Licitra, G

    2014-12-01

    The influence of stage of lactation and corresponding diets on rates of protein degradation (kd) is largely unstudied. Study objectives were to measure and compare in situ ruminal kd of crude protein (CP) and estimate rumen CP escape (rumen-undegradable protein; RUP) of selected feeds by cows at 3 stages of lactation fed corresponding diets, and to determine the incubation times needed in an enzymatic in vitro procedure, using 0.2 units of Streptomyces griseus protease per percent of true CP, that predicted in situ RUP. Residue CP was measured after in situ fermentation for 4, 8, 12, 24, 36, 48, and 72 h of 5 protein sources and 3 total mixed rations, which were fed to the in situ cows. Two nonlactating (dry) cows and 2 cows each at 190 (mid) and 90 (peak) days of lactation were used. Each pair of cows was offered free-choice diets that differed in composition to meet their corresponding nutrient requirements. Diets had decreasing proportions of forages and contained (dry matter basis) 11.9, 15.1 and 16.4% CP and 54.3, 40.3 and 35.3% neutral detergent fiber, for dry, mid, and peak TMR (TMR1, TMR2, and TMR3), respectively. Intakes were 10.3, 21.4, and 23.8kg of dry matter/d, respectively. Kinetic CP fractions (extractable, potentially degradable, undegradable, or slowly degradable) were unaffected by treatment. Lag time and kd varied among feeds. The kd was faster for all feeds (0.136/h) when incubated in dry-TMR1 cows compared with mid-TMR2 (0.097/h) or peak-TMR3 (0.098/h) cows, and no differences in lag time were detected. Calculated RUP, using estimated passage rates for each cow based on intake, differed between dry-TMR1 (0.382) and mid-TMR2 (0.559) or peak-TMR3 (0.626) cows, with a tendency for mid-TMR2 to be different from peak-TMR3. Using the average kd and lag time obtained from dry-TMR1 to calculate RUP for mid-TMR2 and peak-TMR3 cows using their passage rates reduced RUP values by 6.3 and 9.5 percentage units, respectively. Except for that of herring meal

  16. Negative regulation of parathyroid hormone-related protein expression by steroid hormones

    SciTech Connect

    Kajitani, Takashi; Tamamori-Adachi, Mimi; Okinaga, Hiroko; Chikamori, Minoru; Iizuka, Masayoshi; Okazaki, Tomoki

    2011-04-15

    Highlights: {yields} Steroid hormones repress expression of PTHrP in the cell lines where the corresponding nuclear receptors are expressed. {yields} Nuclear receptors are required for suppression of PTHrP expression by steroid hormones, except for androgen receptor. {yields} Androgen-induced suppression of PTHrP expression appears to be mediated by estrogen receptor. -- Abstract: Elevated parathyroid hormone-related protein (PTHrP) is responsible for humoral hypercalcemia of malignancy (HHM), which is of clinical significance in treatment of terminal patients with malignancies. Steroid hormones were known to cause suppression of PTHrP expression. However, detailed studies linking multiple steroid hormones to PTHrP expression are lacking. Here we studied PTHrP expression in response to steroid hormones in four cell lines with excessive PTHrP production. Our study established that steroid hormones negatively regulate PTHrP expression. Vitamin D receptor, estrogen receptor {alpha}, glucocorticoid receptor, and progesterone receptor, were required for repression of PTHrP expression by the cognate ligands. A notable exception was the androgen receptor, which was dispensable for suppression of PTHrP expression in androgen-treated cells. We propose a pathway(s) involving nuclear receptors to suppress PTHrP expression.

  17. Identification of proteins interacting with lactate dehydrogenase in claw muscle of the porcelain crab Petrolisthes cinctipes

    PubMed Central

    Cayenne, Andrea P.; Gabert, Beverly; Stillman, Jonathon H.

    2011-01-01

    Biochemical adaptation of enzymes involves conservation of activity, stability and affinity across a wide range of intracellular and environmental conditions. Enzyme adaptation by alteration of primary structure is well known, but the roles of protein-protein interactions in enzyme adaptation are less well understood. Interspecific differences in thermal stability of lactate dehydrogenase (LDH) in porcelain crabs (genus Petrolisthes) are related to intrinsic differences among LDH molecules and by interactions with other stabilizing proteins. Here, we identified proteins that interact with LDH in porcelain crab claw muscle tissue using co-immunoprecipitation, and showed LDH exists in high molecular weight complexes using size exclusion chromatography and Western blot analyses. Co-immunoprecipitated proteins were separated using 2D SDS PAGE and analyzed by LC/ESI using peptide MS/MS. Peptide MS/MS ions were compared to an EST database for Petrolisthes cinctipes to identify proteins. Identified proteins included cytoskeletal elements, glycolytic enzymes, a phosphagen kinase, and the respiratory protein hemocyanin. Our results support the hypothesis that LDH interacts with glycolytic enzymes in a metabolon structured by cytoskeletal elements that may also include the enzyme for transfer of the adenylate charge in glycolytically produced ATP. Those interactions may play specific roles in biochemical adaptation of glycolytic enzymes. PMID:21968246

  18. Biologically active breast milk proteins in association with very preterm delivery and stage of lactation.

    PubMed

    Mehta, R; Petrova, A

    2011-01-01

    The aim of this study was to identify the independent effect of very preterm gestation on breast milk content of biologically active proteins (secretory immunoglobulin A (sIgA), lysozyme, lactoferrin, osteoprotegerin (OPG), leptin, adiponectin and β-endorphin (b-EP)) during the first month of lactation. We collected samples of transitional (6 to 8 and 13 to 15 days) and mature (20 to 22 and 27 to 29 days) milk from mothers after term (38 to 41 weeks) or very preterm (24 to 31 weeks) delivery. The levels of sIgA, lysozyme, lactoferrin, OPG, leptin, adiponectin and b-EP in the breast milk were quantified using enzyme-linked immunosorbent assay or enzyme immunoassay kits. Statistical analysis included descriptive statistics and regression analysis. Sixty breast milk samples were collected from 15 mothers after very preterm (preterm breast milk, PBM) and 20 samples from 5 mothers after term (term breast milk, TBM) deliveries. Decrease in lysozyme, lactoferrin, OPG, leptin, adiponectin and b-EP but no change in sIgA was recorded during the first month of lactation in both TBM and PBM. The IgA, lysozyme and adiponectin were higher in PBM than in TBM, whereas concentrations of lactoferrin, OPG and leptin were higher in TBM than in PBM (P<0.05 to 0.0001). A similar pattern was seen in the lysozyme, leptin and adiponectin concentration in mature milk. Increased b-EP levels in breast milk were associated with the vaginal mode of delivery but not gestational age. Although a similar pattern of change was observed in the breast milk bioactive proteins during the first month of lactation after term and very preterm gestation, PBM is a better source of factors with antibacterial/anti-inflammatory activities but is constantly deficient in leptin, which is involved in neuroendocrine regulation.

  19. Modelling lactation curve for milk fat to protein ratio in Iranian buffaloes (Bubalus bubalis) using non-linear mixed models.

    PubMed

    Hossein-Zadeh, Navid Ghavi

    2016-08-01

    The aim of this study was to compare seven non-linear mathematical models (Brody, Wood, Dhanoa, Sikka, Nelder, Rook and Dijkstra) to examine their efficiency in describing the lactation curves for milk fat to protein ratio (FPR) in Iranian buffaloes. Data were 43 818 test-day records for FPR from the first three lactations of Iranian buffaloes which were collected on 523 dairy herds in the period from 1996 to 2012 by the Animal Breeding Center of Iran. Each model was fitted to monthly FPR records of buffaloes using the non-linear mixed model procedure (PROC NLMIXED) in SAS and the parameters were estimated. The models were tested for goodness of fit using Akaike's information criterion (AIC), Bayesian information criterion (BIC) and log maximum likelihood (-2 Log L). The Nelder and Sikka mixed models provided the best fit of lactation curve for FPR in the first and second lactations of Iranian buffaloes, respectively. However, Wood, Dhanoa and Sikka mixed models provided the best fit of lactation curve for FPR in the third parity buffaloes. Evaluation of first, second and third lactation features showed that all models, except for Dijkstra model in the third lactation, under-predicted test time at which daily FPR was minimum. On the other hand, minimum FPR was over-predicted by all equations. Evaluation of the different models used in this study indicated that non-linear mixed models were sufficient for fitting test-day FPR records of Iranian buffaloes.

  20. Leptin and prolactin, but not corticosterone, modulate body weight and thyroid function in protein-malnourished lactating rats.

    PubMed

    Lisboa, P C; Passos, M C; Dutra, S C; Bonomo, I T; Denolato, A T; Reis, A M; Moura, E G

    2006-05-01

    To understand the role of hormonal changes in the lower food ingestion and body weight in protein-restricted lactating rats as well as the higher serum T (3), higher deiodination, iodide and T (3) milk transfer, we measured maternal serum prolactin, leptin, TSH and corticosterone, which are hormones that could influence those parameters. After birth, dams were separated into: control-fed with a 23 % protein diet (n = 12) and PR (protein-restricted)-fed with an 8 % protein diet (n = 12). At the 4 (th) and 21 (st) day of lactation, half of the animals in each group were sacrificed. PR dams presented hyperleptinemia (day 4: + 20 %; day 21: + 19 %; p < 0.05) and hypoprolactinemia (day 4: - 85 %; day 21: - 92 %; p < 0.05), which could help explain the lower food consumption and body weight in lactating PR rats since leptin is anorexigenic and prolactin is orexigenic. Also, this hyperleptinemia could contribute for the increase in serum T (3) of PR dams, since leptin stimulates T (3) production, especially acting on deiodinases. Serum corticosterone was not different between PR and C groups, and TSH was lower only at the end of lactation. Thus, we suggest that both leptin and prolactin could play an important role in the body weight and thyroid hormone changes observed in protein-malnourished lactating rats.

  1. A maternal low protein diet during pregnancy and lactation in the rat impairs male reproductive development

    PubMed Central

    Zambrano, E; Rodríguez-González, GL; Guzmán, C; García-Becerra, R; Boeck, L; Díaz, L; Menjivar, M; Larrea, F; Nathanielsz, PW

    2005-01-01

    Nutrient restriction during pregnancy and lactation impairs growth and development. Recent studies demonstrate long-term programming of function of specific organ systems resulting from suboptimal environments during fetal life and development up to weaning. We determined effects of maternal protein restriction (50% control protein intake) during fetal development and/or lactation in rats on the reproductive system of male progeny. Rats were fed either a control 20% casein diet (C) or a restricted diet (R) of 10% casein during pregnancy. After delivery mothers received either C or R diet until weaning to provide four groups: CC, RR, CR and RC. We report findings in male offspring only. Maternal protein restriction increased maternal serum corticosterone, oestradiol and testosterone (T) concentrations at 19 days gestation. Pup birth weight was unchanged but ano-genital distance was increased by maternal protein restriction (P < 0.05). Testicular descent was delayed 4.4 days in RR, 2.1 days in CR and 2.2 days in RC and was not related to body weight. Body weight and testis weight were reduced in RR and CR groups at all ages with the exception of CR testis weight at 270 days postnatal age (PN). At 70 days PN luteinizing hormone and T concentrations were reduced in RR, CR and RC. mRNA for P450 side chain cleavage (P450scc) was reduced in RR and CR at 21 days PN but was unchanged at 70 days PN. Fertility rate was reduced at 270 days PN in RC and sperm count in RR and RC. We conclude that maternal protein delays sexual maturation in male rats and that some effects only emerge in later life. PMID:15611025

  2. Lactate dehydrogenase test

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003471.htm Lactate dehydrogenase test To use the sharing features on this page, please enable JavaScript. Lactate dehydrogenase (LDH) is a protein that helps produce energy ...

  3. Adipose tissue angiopoietin-like protein 4 messenger RNA changes with altered energy balance in lactating Holstein cows.

    PubMed

    Koltes, D A; Spurlock, D M

    2012-11-01

    Negative energy balance at the onset of lactation is unfavorably associated with fitness traits in high-producing dairy cows. Angiopoietin-like protein 4 (ANGPTL4) is an adipokine that has been associated with the regulation of lipid metabolism through the inhibition of lipoprotein lipase activity and regulation of lipolysis. Expression of ANGPTL4 messenger RNA (mRNA) increases during early lactation, but its regulation with changing energy status is currently unknown. Accordingly, the objective of this study was to determine whether ANGPTL4 mRNA abundance is responsive to declining energy balance induced by the transition from pregnancy to lactation, feed restriction, and GH administration in lactating dairy cows. The mRNA abundance of leptin, adiponectin, and adiponectin receptor 2 were also measured to compare adipokine mRNA profiles during changes in energy metabolism. Repeated adipose tissue biopsies were taken from different cows during transition from late pregnancy to lactation (n = 26), feed restriction (n = 19), and GH administration (n = 20). As expected, milk yield increased with the onset of lactation and GH administration (P < 0.01) but declined during feed restriction. Energy balance declined in each experiment, resulting in negative energy balance at the onset of lactation and after feed restriction. Abundance of ANGPTL4 mRNA expression increased 2- to 6-fold with declining energy balance in each experiment. Leptin mRNA declined with feed restriction, and adiponectin mRNA decreased with the onset of lactation. The consistency and magnitude of the increase in ANGPTL4 mRNA across multiple models of altered energy balance identifies it as an adipokine that is uniquely responsive to changes in energy balance in the lactating dairy cow. Copyright © 2012 Elsevier Inc. All rights reserved.

  4. [Hypercalcemia related to PTH-rP revealing malignant hepatic epithelioid hemangioendothelioma].

    PubMed

    Boukhris, Imène; Azzabi, Samira; Kéchaou, Ines; Chérif, Eya; Kooli, Chékib; Romdhane, Khaled Ben; Omar, Souheil; Khalfallah, Narjes

    2016-01-01

    Hypercalcemia caused by tumor production of PTH-rp occurs most often in cases of squamous cell carcinoma of the lung, aerodigestive tract cancer, gynecological cancer and lymphoma. We report an exceptional case of PTH-rp related to a hepatic hemangioendothelioma. A 70 years-old male admitted for deterioration of the general state. The laboratory investigations revealed hypercalcemia, related to tumor production of PTH-rp. Imaging revealed tumoral hepatic lesions. Histopathological study and immunohistochemistry showed diffuse response for CD31 marker, CK20 (+) with CK7 (-) and hepatocyt antigen (-). The diagnosis of PTH-rp related to hepatic hemangioendothelioma was make. The patient died with recurrence of fatal hypercalcemia. Management of patients presenting with humoral hypercalcemia includes a vigorous search for tumor lesions. Elevated PTH-rp can be a bad prognostic factor. In front of tumoral liver lesions, a hepatic epithelioid hemangioendothelioma must be considered. Immunohistochemistry is necessary to make diagnosis.

  5. Lactation Affects Isolated Mitochondria and Its Fatty Acid Composition but Has No Effect on Tissue Protein Oxidation, Lipid Peroxidation or DNA-Damage in Laboratory Mice

    PubMed Central

    Valencak, Teresa G.; Raith, Johannes; Staniek, Katrin; Gille, Lars; Strasser, Alois

    2016-01-01

    Linking peak energy metabolism to lifespan and aging remains a major question especially when focusing on lactation in females. We studied, if and how lactation affects in vitro mitochondrial oxygen consumption and mitochondrial fatty acid composition. In addition, we assessed DNA damage, lipid peroxidation and protein carbonyls to extrapolate on oxidative stress in mothers. As model system we used C57BL/6NCrl mice and exposed lactating females to two ambient temperatures (15 °C and 22 °C) while they nursed their offspring until weaning. We found that state II and state IV respiration rates of liver mitochondria were significantly higher in the lactating animals than in non-lactating mice. Fatty acid composition of isolated liver and heart mitochondria differed between lactating and non-lactating mice with higher n-6, and lower n-3 polyunsaturated fatty acids in the lactating females. Surprisingly, lactation did not affect protein carbonyls, lipid peroxidation and DNA damage, nor did moderate cold exposure of 15 °C. We conclude that lactation increases rates of mitochondrial uncoupling and alters mitochondrial fatty acid composition thus supporting the “uncoupling to survive” hypothesis. Regarding oxidative stress, we found no impact of lactation and lower ambient temperature and contribute to growing evidence that there is no linear relationship between oxidative damage and lactation. PMID:26805895

  6. Lactate dehydrogenase and alkaline phosphatase isoenzymes and protein-bound sialic acid in regenerating rat liver.

    PubMed

    Allalouf, D; Schwarzman, S; Levinsky, H; Feller, N; Hart, J; Zoher, S; Menache, R

    1986-01-01

    Lactate dehydrogenase (LDH) and alkaline phosphatase (AP) isoenzyme patterns and protein-bound sialic acid content were compared between normal, regenerating rat liver 10 days after partial hepatectomy and fetal rat liver. For this purpose, liver from ten adult rats and two pools of ten fetal livers each were examined. Isoenzymes were separated by electrophoresis on cellulose acetate and their percent distribution calculated after quantitation by densitometry of the bands. LDH-5 and LDH-4 combined represented in all the tissues examined 90%-94% of the total activity. LDH-5/LDH-4 ratios were nearly equivalent in the normal and regenerated liver (7.14, 6.41), but substantially lower in fetal liver (2.50). Two bands of AP were visualized in electropherograms. AP-1/AP-2 ratio was lower in regenerated liver (1.57) as compared to normal liver (2.27) and still lower in fetal liver (1.06). Protein-bound sialic acid was, on protein basis, slightly but not significantly higher in regenerated liver (1.71 microgram/mg protein) than in normal liver (1.43), and significantly higher in fetal liver (1.87). The relatively small differences in isoenzyme patterns and in protein-bound sialic acid between regenerated and normal liver as compared to those between fetal and normal tissue add support to the view that the cells in regenerated liver are not of embryonic origin.

  7. Protein deficiency alters impact of intestinal nematode infection on intestinal, visceral and lymphoid organ histopathology in lactating mice.

    PubMed

    Starr, Lisa M; Odiere, Maurice R; Koski, Kristine G; Scott, Marilyn E

    2014-05-01

    Protein deficiency impairs local and systemic immune responses to Heligmosomoides bakeri infection but little is known about their individual and interactive impacts on tissue architecture of maternal lymphoid (thymus, spleen) and visceral (small intestine, kidney, liver, pancreas) organs during the demanding period of lactation. Using a 2 × 2 factorial design, pregnant CD1 mice were fed a 24% protein sufficient (PS) or a 6% protein deficient (PD) isoenergetic diet beginning on day 14 of pregnancy and were infected with 100 H. bakeri larvae four times or exposed to four sham infections. On day 20 of lactation, maternal organs were examined histologically and serum analytes were assayed as indicators of organ function. The absence of villus atrophy in response to infection was associated with increased crypt depth and infiltration of mast cells and eosinophils but only in lactating dams fed adequate protein. Infection-induced lobular liver inflammation was reduced in PD dams, however, abnormalities in the kidney caused by protein deficiency were absent in infected dams. Bilirubin and creatinine were highest in PD infected mice. Infection-induced splenomegaly was not due to an increase in the lymphoid compartment of the spleen. During lactation, infection and protein deficiency have interactive effects on extra-intestinal pathologies.

  8. An Herbal Galactagogue Mixture Increases Milk Production and Aquaporin Protein Expression in the Mammary Glands of Lactating Rats

    PubMed Central

    Liu, Haibin; Hua, Ying; Luo, Hui; Shen, Zhaojun; Tao, Xuejiao

    2015-01-01

    Background. Herbal galactagogues have been increasingly used to treat postpartum hypogalactia. The mechanism of action of herbal galactagogues remains unclear. The purpose of this study was to investigate the effect of an herbal galactagogue mixture on milk production and aquaporin (AQP) expression in lactating rats. Methods. Thirty female Sprague Dawley rats were randomized into virgin, lactating + H2O, and lactating + galactagogue groups (n = 10 per group). Lactating rats were administered the decoction of an herbal galactagogue mixture by oral gavage or the same amount of distilled water. Results. The herbal decoction significantly increased milk production in lactating rats (P < 0.05). Both immunohistochemical staining and western blot showed that protein levels of AQP-3 and AQP-5 were significantly increased during lactation compared with virgin stage and the herbal decoction further elevated their expression (P < 0.05). AQP-1 was predominantly expressed in the capillaries whereas AQP-3 and AQP-5 were mainly detected in the epithelial cells and ducts of the mammary glands. Conclusion. The expression of AQPs in the mammary glands of rats was developmentally regulated. Herbal galactagogues might have increased milk secretion by regulating the expression and function of AQPs in the mammary glands. PMID:26075000

  9. The lactate receptor, G-protein-coupled receptor 81/hydroxycarboxylic acid receptor 1: Expression and action in brain.

    PubMed

    Morland, Cecilie; Lauritzen, Knut Husø; Puchades, Maja; Holm-Hansen, Signe; Andersson, Krister; Gjedde, Albert; Attramadal, Håvard; Storm-Mathisen, Jon; Bergersen, Linda Hildegard

    2015-07-01

    We have proposed that lactate is a "volume transmitter" in the brain and underpinned this by showing that the lactate receptor, G-protein-coupled receptor 81 (GPR81, also known as HCA1 or HCAR1), which promotes lipid storage in adipocytes, is also active in the mammalian brain. This includes the cerebral neocortex and the hippocampus, where it can be stimulated by physiological concentrations of lactate and by the HCAR1 agonist 3,5-dihydroxybenzoate to reduce cAMP levels. Cerebral HCAR1 is concentrated on the postsynaptic membranes of excitatory synapses and also is enriched at the blood-brain barrier. In synaptic spines and in adipocytes, HCAR1 immunoreactivity is also located on subplasmalemmal vesicular organelles, suggesting trafficking to and from the plasma membrane. Through activation of HCAR1, lactate can act as a volume transmitter that links neuronal activity, cerebral blood flow, energy metabolism, and energy substrate availability, including a glucose- and glycogen-saving response. HCAR1 may contribute to optimizing the cAMP concentration. For instance, in the prefrontal cortex, excessively high cAMP levels are implicated in impaired cognition in old age, fatigue, stress, and schizophrenia and in the deposition of phosphorylated tau protein in Alzheimer's disease. HCAR1 could serve to ameliorate these conditions and might also act through downstream mechanisms other than cAMP. Lactate exits cells through monocarboxylate transporters in an equilibrating manner and through astrocyte anion channels activated by depolarization. In addition to locally produced lactate, lactate produced by exercising muscle as well as exogenous HCAR1 agonists, e.g., from fruits and berries, might activate the receptor on cerebral blood vessels and brain cells.

  10. Identification and functional characterization of a novel monotreme- specific antibacterial protein expressed during lactation.

    PubMed

    Bisana, Swathi; Kumar, Satish; Rismiller, Peggy; Nicol, Stewart C; Lefèvre, Christophe; Nicholas, Kevin R; Sharp, Julie A

    2013-01-01

    Monotremes are the only oviparous mammals and exhibit a fascinating combination of reptilian and mammalian characters. They represent a component of synapsidal reproduction by laying shelled eggs which are incubated outside the mother's body. This is accompanied by a prototherian lactation process, marking them as representatives of early mammals. The only extant monotremes are the platypus, and the short- and long- beaked echidnas, and their distributions are limited to Australia and New Guinea. Apart for a short weaning period, milk is the sole source of nutrition and protection for the hatchlings which are altricial and immunologically naive. The duration of lactation in these mammals is prolonged relative to the gestational length and period of incubation of eggs. Much of the development of monotreme young occurs in the non-sterile ex-utero environment. Therefore the role of milk in the growth, development and disease protection of the young is of significant interest. By sequencing the cDNA of cells harvested from monotreme milk, we have identified a novel monotreme- specific transcript, and the corresponding gene was designated as the EchAMP. The expression profile of this gene in various tissues revealed that it is highly expressed in milk cells. The peptides corresponding to the EchAMP protein have been identified in a sample of echidna milk In silico analysis indicated putative antimicrobial potential for the cognate protein of EchAMP. This was further confirmed by in vitro assays using a host of bacteria. Interestingly, EchAMP did not display any activity against a commensal gut floral species. These results support the hypothesis of enhancement of survival of the young by antimicrobial bioactives of mammary gland origin and thus emphasize the protective, non- nutritional role of milk in mammals.

  11. Application of capillary zone electrophoresis to the characterisation of the human milk protein profile and its evolution throughout lactation.

    PubMed

    Manso, M A; Miguel, M; López-Fandiño, R

    2007-03-30

    This work describes the use of capillary zone electrophoresis for the characterisation of human milk proteins. The major proteins were identified following different strategies, such as the treatment with enzymes for selective protein modification. Using this method we studied the proteins in human milk from different donors throughout lactation. Qualitative and quantitative differences in the composition of the individual proteins were observed. The different beta-casein phosphoforms were separated and quantified. The average proportion of the 0P:1P:2P:3P:4P:5P was, approximately, 3:6:9:4:10:2. The evolution of the ratio of the different beta-casein phosphoforms during lactation is reported.

  12. Effects of dry period length and concentrate protein content in late lactation on body condition score change and subsequent lactation performance of thin high genetic merit dairy cows.

    PubMed

    Dale, A J; Purcell, P J; Wylie, A R G; Gordon, A W; Ferris, C P

    2017-03-01

    Improving body condition score of thin cows in late lactation is necessary, because cows that are thin at drying off exhibit decreased fertility postpartum and are at increased risk of disease and of being culled in the subsequent lactation. Offering a diet low in crude protein (CP) content in late lactation may help to improve body condition score (BCS) at drying off, whereas imposing an extended dry period (EDP) has been advocated as another way to increase BCS at calving. To test these hypotheses, 65 thin cows (mean BCS 2.25 at 14 wk precalving) were managed on 1 of 3 treatments between 13 and 9 wk prepartum: normal protein control {NP; grass silage + 5 kg/d of a normal protein concentrate [228 g of CP/kg of dry matter (DM)]}, low protein [LP; grass silage + 5 kg/d of a low-protein concentrate (153 g of CP/kg of DM)], or EDP (cows dried off at 13 wk precalving and offered a grass silage-only diet). Both NP and LP cows were dried off at wk 8 prepartum, after which all cows were offered a grass silage-only diet until calving. After calving, all cows were offered a common diet (supplying 11.1 kg of concentrate DM/cow per day) for 19 wk. Between 13 and 9 wk prepartum, LP cows had lower DM intake, milk yield, and body weight than NP cows. Whereas EDP cows had lower serum β-hydroxybutyrate and fatty acid concentrations than those of NP cows, BCS at wk 9 prepartum did not differ between treatments. Cows on the LP treatment continued to have lower DMI and BW than those of NP and EDP cows between 8 wk prepartum and calving, but only EDP cows had a higher BCS at calving. Treatment did not affect calving difficulty score or calf birth weight. Although all cows were offered a common diet postpartum, cows on the LP treatment had lower DM intake and milk fat + plus protein yield than cows on any other treatment during the 19-wk period postpartum, but we found no differences in any postpartum indicator of body tissue reserves. The treatments imposed from wk 13 to 9 prepartum

  13. Single cell protein from wood pulp waste as a feed supplement for lactating cows

    SciTech Connect

    Claypool, D.W.; Church, D.C.

    1984-01-01

    Twenty-four Holstein cows were assigned to one of three rations at the beginning of their first or second lactation and were kept on the rations for 16 wk. Rations differed only that a single cell protein material marketed under the name RayPro (ITT Rayonier, Inc., trademark) was substituted for cottonseed meal at 0, 3, and 7% (dry matter) of the total ration. Crude protein analyses were 15.6, 15.9, and 16.5% for rations containing 0, 3, and 7% RayPro. Rations were composed of 50% concentrate, 25% chopped alfalfa hay, and 25% corn silage dry matter. Milk fat percentages were 3.2, 3.6, and 3.7% for rations with 0, 3, and 7% RayPro. Cows fed the ration containing 7% RayPro had higher rumen ammonia (6.0, 5.9, 8.1 mg/dl) and higher urea in blood plasma (12.7, 14.9, 16.3 mg/dl) than cows fed the 0 and 3% RayPro rations. RayPro is an acceptable substitute for conventional protein supplements in amounts used in this trial when fed as part of a completely mixed ration. 12 references, 2 tables.

  14. The effect of sodium lactate, potassium lactate, carrageenan, whey protein concentrate, yeast extract and fungal proteinases on the cook yield and tenderness of bovine chuck muscles.

    PubMed

    Walsh, H; Martins, S; O'Neill, E E; Kerry, J P; Kenny, T; Ward, P

    2010-06-01

    This study evaluated the efficacy of injection with enhancement solutions containing sodium lactate (NaLac), potassium lactate (KLac), carrageenan, whey protein concentrate (WPC), yeast extract or fungal proteinases alone or in combination with NaCl at increasing the tenderness and cook yield of bovine M.supraspinatus and M. tricepsbrachii caput longum. Muscle sections (400g) were injected with enhancement solutions at an injection rate of 110% to give specific residual concentrations as follows: 0.5% NaCl; 2% NaLac; 2% KLac; 0.5% NaCl+1 or 2% NaLac; 0.5% NaCl+1% KLac; 0.5% NaCl+0.4% sodium tripolyphosphate (STPP)+2% NaLac; 0.5% NaCl+0.4% STPP, 1.5% WPC+/-0.5% NaCl, 1.0% carrageenan+/-0.5% NaCl, 0.5% yeast extract or 0.0005% and 0.001% fungal enzymes. Injection with solutions containing sodium or potassium lactates (alone or in combination with NaCl and STPP), WPC, carrageenan and yeast extract significantly (P<0.05) decreased WBSF values and increased sensory tenderness ratings, compared to non-injected controls. All of these ingredients in water also significantly (P<0.05) increased cook yield with the exception of yeast extract which had no significant effect. The fungal enzymes in a water carrier acted as effective tenderising agents in both muscles but when used at the higher level resulted in a significant decrease in percentage cook yield. Injection of the chuck muscles with these ingredients and water represents an effective method of improving their tenderness and in most cases also increasing percentage cook yield.

  15. Changes over lactation in breast milk serum proteins involved in the maturation of immune and digestive system of the infant.

    PubMed

    Zhang, Lina; de Waard, Marita; Verheijen, Hester; Boeren, Sjef; Hageman, Jos A; van Hooijdonk, Toon; Vervoort, Jacques; van Goudoever, Johannes B; Hettinga, Kasper

    2016-09-16

    To objective of this study was to better understand the biological functions of breast milk proteins in relation to the growth and development of infants over the first six months of life. Breast milk samples from four individual women collected at seven time points in the first six months after delivery were analyzed by filter aided sample preparation and dimethyl labeling combined with liquid chromatography tandem mass spectrometry. A total of 247 and 200 milk serum proteins were identified and quantified, respectively. The milk serum proteome showed a high similarity (80% overlap) on the qualitative level between women and over lactation. The quantitative changes in milk serum proteins were mainly caused by three groups of proteins, enzymes, and transport and immunity proteins. Of these 21 significantly changed proteins, 30% were transport proteins, such as serum albumin and fatty acid binding protein, which are both involved in transporting nutrients to the infant. The decrease of the enzyme bile salt-activated lipase as well as the immunity proteins immunoglobulins and lactoferrin coincide with the gradual maturation of the digestive and immune system of infants. The human milk serum proteome didn't differ qualitatively but it did quantitatively, both between mothers and as lactation advanced. The changes of the breast milk serum proteome over lactation corresponded with the development of the digestive and immune system of infants. Breast milk proteins provide nutrition, but also contribute to healthy development of infants. Despite the previously reported large number of identified breast milk proteins and their changes over lactation, less is known on the changes of these proteins in individual mothers. This study is the first to determine the qualitative and quantitative changes of milk proteome over lactation between individual mothers. The results indicate that the differences in the milk proteome between individual mothers are more related to the

  16. Effect of prepartum energy intake and postpartum protein source on plasma somatotropin and insulin in lactating cows.

    PubMed

    Seymour, W M; Herbein, J H; Akers, R M; Polan, C E

    1988-11-01

    Forty-two Holstein cows were fed a high or low energy diet during the last third of lactation, then fed soybean meal or dried brewers grains in the subsequent lactation. Responses of plasma somatotropin and insulin to arginine were measured at 6 and 15 wk postpartum. Basal somatotropin, response area, and peak value were greater at 6 versus 15 wk of lactation. Insulation responses were greater at 15 wk of lactation. At 6 wk, basal somatotropin was higher (6.1 versus 4.1 ng/ml) in cows fed the low energy diet prepartum. Shape of somatotropin response curves at 6 wk differed between prepartum energy intakes, but response areas were similar. Plasma insulin at 6 wk tended to be higher (1.0 versus .7 ng/ml) in cows fed dried brewers grains. At 15 wk, both insulin and somatotropin response areas tended to be smaller for cows fed dried brewers grains, and shape of insulin response curves differed between protein sources. Of the observed hormonal responses to diet, only 6-wk plasma somatotropin was associated with differences in production parameters. Somatotropin secretion during early lactation appears to respond to prepartum changes in the energy status.

  17. Replacing alfalfa hay with dry corn gluten feed and Chinese wild rye grass: Effects on rumen fermentation, rumen microbial protein synthesis, and lactation performance in lactating dairy cows.

    PubMed

    Hao, X Y; Gao, H; Wang, X Y; Zhang, G N; Zhang, Y G

    2017-04-01

    This experiment was conducted to investigate nutrient digestibility, rumen microbial protein synthesis, and lactation performance when a portion of alfalfa was replaced with combinations of dry corn gluten feed (DCGF) and Chinese wild rye grass in the diet of lactating cows. Six multiparous and 3 primiparous Chinese Holsteins were arranged in a replicated 3 × 3 Latin square experiment for 21-d periods. The animals were fed 1 of 3 treatment diets during each period: (1) 0% DCGF (0DCGF); (2) 6.5% DCGF (7DCGF); and (3) 11% DCGF (11DCGF). Diets were isonitrogenous, and a portion of alfalfa hay was replaced with DCGF and Chinese wild rye grass, with similar concentrate mixtures and corn silage contents. The dry matter intake was greater for 11DCGF (21.9 kg/d) than for 0DCGF (20.7 kg/d) or 7DCGF (21.2 kg/d). The treatment diets did not result in difference in milk production, fat and lactose concentration, or yield. Compared with 0DCGF, the ration containing 11% DCGF improved the milk protein concentration. Dry matter and neutral detergent fiber digestibility was greater for 7DCGF (62.7% and 45.6%) and 11DCGF (63.1% and 47.2%) than for 0DCGF (59.4% and 42.3%), and the nitrogen digestibility was similar for the 3 treatments. The concentration of rumen volatile fatty acids was higher in cows fed the 11DCGF diet than in those fed the 0DCGF diet, with no difference between the 7DCGF and 11DCGF diets. The estimated microbial crude protein yield was greater for the 11DCGF diet (1985.1 g/d) than for the 0DCGF diet (1745.0 g/d), with no difference between the 0DCGF and 7DCGF diets. Thus, it appears that feeding DCGF and Chinese wild rye grass in combination can effectively replace a portion of alfalfa hay in the rations of lactating dairy cows.

  18. Transfer of protein antigens into milk after intravenous injection into lactating mice

    SciTech Connect

    Harmatz, P.R.; Hanson, D.G.; Walsh, M.K.; Kleinman, R.E.; Bloch, K.J.; Walker, W.A.

    1986-08-01

    We investigated the transfer of bovine serum /sup 125/I-albumin (/sup 125/I-BSA), bovine /sup 125/I-gamma-globulin (/sup 125/I-BGG), /sup 125/I-ovalbumin (/sup 125/I-OVA), and /sup 125/I-beta-lactoglobulin (/sup 125/I-BLG) from the blood into the milk of lactating mice. Equal amounts (by weight) of the radiolabeled proteins were injected intravenously into mice 1 wk postpartum. Total radioactivity, trichloroacetic acid-precipitable radioactivity, and specifically immunoprecipitable radioactivity were measured in serum, mammary gland homogenate, and milk. Clearance of immunoreactive OVA (iOVA) and iBLG from the circulation was more rapid than iBSA and iBGG. The radioactivity in mammary tissue associated with BSA and BGG was greater than 70% immunoprecipitable throughout the 4-h test interval; /sup 125/I-OVA and /sup 125/I-BLG were less than 12% precipitable 1 and 4 h after injection. In milk obtained at 4 h, there was an approximately 10-fold greater accumulation of iBSA or iBGG than of iOVA or iBLG. These experiments demonstrate that protein antigens differ in their ability to transfer from maternal circulation into milk. The transfer into milk appeared to be in proportion to persistence of the antigens in the maternal circulation.

  19. Copper transport during lactation in transgenic mice expressing the human ATP7A protein

    PubMed Central

    Llanos, Roxana M.; Michalczyk, Agnes A.; Freestone, David J.; Currie, Scott; Linder, Maria C.; Ackland, M. Leigh; Mercer, Julian F.B.

    2008-01-01

    Both copper transporting ATPases, ATP7A and ATP7B, are expressed in mammary epithelial cells but their role in copper delivery to milk has not been clarified. We investigated the role of ATP7A in delivery of copper to milk using transgenic mice that over-express human ATP7A. In mammary gland of transgenic mice, human ATP7A protein was 10- to 20-fold higher than in control mice, and was localized to the basolateral membrane of mammary epithelial cells in lactating mice. The copper concentration in the mammary gland of transgenic dams and stomach contents of transgenic pups was significantly reduced compared to non-transgenic mice. The mRNA levels of endogenous Atp7a, Atp7b, and Ctr1 copper transporters in the mammary gland were not altered by the expression of the ATP7A transgene, and the protein levels of Atp7b and ceruloplasmin were similar in transgenic and non-transgenic mice. These data suggest that ATP7A plays a role in removing excess copper from the mammary epithelial cells rather than supplying copper to milk. PMID:18515074

  20. [Blood concentrations of lactate, C-reactive protein, and creatinine as early indicators of severity and outcome of sepsis].

    PubMed

    Mikić, Dragan; Arsić-Komljenović, Gordana; Nozić, Darko; Cućuz, Milica; Dimitrijević, Radmila; Vukadinov, Jovan

    2010-01-01

    Since the outcome in septic patients can significantly be improved if the appropriate therapy is introduced timely early, the early diagnosis of sepsis and its complications is essential. The aim of this study was to compare mean values of the initial blood concentrations of lactate, C-reactive protein and creatinine and the severity of illness and the outcome of sepsis. A total of 30 septic patients were included in the study. The diagnosis of sepsis and its complications was made according to consensus criteria. The severity of illness was scored by an acute physiology, age and chronic health evaluation septic score. The patients were subdivided into different groups, those with sepsis, severe sepsis or septic shock, those with or without multiple organ dysfunction syndrome, and survivors and nonsurvivors. The differences in mean values of lactate levels among all studied groups were significantly high, whereas the level of C-reactive protein were significantly higher only in the non-survivors compared to the survivors (p < 0.05). The concentrations of creatinine were significantly higher in the patients with septic shock compared to the patients with sepsis, and in the patients with multiple organ dysfunction syndrome and the non-survivors compared to the corresponding groups (p < 0.05). The septic score clearly discriminated patients with different severity of sepsis, development of multiple organ dysfunction syndrome and survival and positively correlated with the concentrations of lactate, C-reactive protein and creatinine (the best correlation ranks were with lactate levels, p < 0.001). Our results suggest that lactate level is a better parameter of illness severity and outcome of sepsis than levels of C-reactive protein and creatinine. When compared to the above parameters, the septic score determined on the day of admission to hospital is a much better criterion to classify patients into groups with different severity of sepsis, with and without multiple

  1. Feeding protein supplements in alfalfa hay-based lactation diets improves nutrient utilization, lactational performance, and feed efficiency of dairy cows.

    PubMed

    Neal, K; Eun, J-S; Young, A J; Mjoun, K; Hall, J O

    2014-12-01

    Due to the increasing cost of soybean meal and concerns of excess N being excreted into the environment, new protein supplements have been developed. Two products that have shown potential in increasing N utilization efficiency are slow-release urea (SRU; Optigen; Alltech Inc., Nicholasville, KY) and ruminal-escape protein derived from yeast (YMP; DEMP; Alltech Inc.). The objective of this study was to assess the effects of feeding these 2 supplements in alfalfa hay-based [45.7% of forage dietary dry matter (DM)] dairy diets on nutrient utilization, feed efficiency, and lactational performance of dairy cows. Twelve multiparous dairy cows were used in a triple 4 × 4 Latin square design with one square consisting of ruminally cannulated cows. Treatments included (1) control, (2) SRU-supplemented total mixed ration (SRUT), (3) YMP-supplemented total mixed ration (YMPT), and (4) SRU- and YMP-supplemented total mixed ration (SYT). The control consisted only of a mixture of soybean meal and canola meal in a 50:50 ratio. The SRU and the YMP were supplemented at 0.49 and 1.15% DM, respectively. The experiment consisted of 4 periods lasting 28 d each (21 d of adaptation and 7 d of sampling). Cows fed YMPT and SYT had decreased intake of DM, and all supplemented treatments had lower crude protein intake compared with those fed the control. Milk yield tended to have the greatest increase in YMPT compared with the control (41.1 vs. 39.7 kg/d) as well as a tendency for increased milk fat and protein yields. Feed efficiencies based on yields of milk, 3.5% fat-corrected milk, and energy-corrected milk increased at 10 to 16% due to protein supplementation. Cows fed protein supplements partitioned less energy toward body weight gain, but tended to partition more energy toward milk production. Efficiency of use of feed N to milk N increased by feeding SRUT and YMPT, and milk N-to-manure N ratio increased with YMPT. Overall results from this experiment indicate that replacing the

  2. Dinosaur lactation?

    PubMed

    Else, Paul L

    2013-02-01

    Lactation is a process associated with mammals, yet a number of birds feed their newly hatched young on secretions analogous to the milk of mammals. These secretions are produced from various sections (crop organ, oesophageal lining and proventriculus) of the upper digestive tract and possess similar levels of fat and protein, as well as added carotenoids, antibodies and, in the case of pigeons and doves, epidermal growth factor. Parental care in avian species has been proposed to originate from dinosaurs. This study examines the possibility that some dinosaurs used secretory feeding to increase the rate of growth of their young, estimated to be similar to that of present day birds and mammals. Dinosaur 'lactation' could also have facilitated immune responses as well as extending parental protection as a result of feeding newly hatched young in nest environments. While the arguments for dinosaur lactation are somewhat generic, a case study for lactation in herbivorous site-nesting dinosaurs is presented. It is proposes that secretory feeding could have been used to bridge the gap between hatching and establishment of the normal diet in some dinosaurs.

  3. Parathyroid-hormone-related protein in sarcoidosis.

    PubMed Central

    Zeimer, H. J.; Greenaway, T. M.; Slavin, J.; Hards, D. K.; Zhou, H.; Doery, J. C.; Hunter, A. N.; Duffield, A.; Martin, T. J.; Grill, V.

    1998-01-01

    Parathyroid-hormone-related protein (PTHrP) is the main mediator of the humoral hypercalcemia of malignancy. It is also detected in many normal adult and fetal tissues. Altered calcium metabolism occurs in sarcoidosis, and two cases of sarcoidosis with hypercalcemia and elevated plasma PTHrP are described. An archival study of 20 lymph node biopsies with the pathological diagnosis of sarcoidosis was performed. Immunohistochemistry using a polyclonal antiserum to human PTHrP and in situ hybridization using a riboprobe to human PTHrP were performed on the lymph node biopsies. Immunohistochemistry for PTHrP was also performed on the biopsies from the two cases with elevated plasma levels. Immunohistochemical analysis detected PTHrP in macrophages within granulomata in 17 of the 20 (85%) biopsies. In situ hybridization detected a positive signal for messenger RNA in the granulomata of 11 of 19 (58%) biopsies. PTHrP immunoreactivity and PTHrP gene expression are present in sarcoid granulomata. PTHrP may contribute to the hypercalcemia of sarcoidosis. Images Figure 1 PMID:9422518

  4. Short communication: Circulating serotonin (5-HT) concentrations on day 1 of lactation as a potential predictor of transition-related disorders.

    PubMed

    Laporta, J; Moore, S A E; Peters, M W; Peters, T L; Hernandez, L L

    2013-08-01

    The monoamine serotonin (5-hydroxytryptamine; 5-HT) has been described as a homeostatic regulator of lactation. Recently, our laboratory determined that 5-HT is involved in the regulation of calcium and glucose homeostasis during the transition period in rodents. More specifically, we demonstrate that 5-HT is responsible for calcium mobilization from bone and upregulation of hepatic gluconeogenic enzymes and mammary gland glucose transporters. Our objective was to investigate the correlation between circulating 5-HT concentrations and circulating ionized calcium, parathyroid hormone-related protein (PTHrP), and glucose concentrations on d 1 postpartum. We also investigated the correlation between circulating 5-HT and milk fever and ketosis incidence and severity in multiparous Holstein cows at the onset of lactation. Blood samples were collected from 42 multiparous cows on d 1 of lactation and analyzed for 5-HT, calcium, glucose, and PTHrP. Milk fever (determined subjectively for each cow on d 1 postpartum) and ketosis incidence and severity (scale 1 to 4, determined objectively for each cow during the first 10 d postpartum) were recorded for all animals. Serum 5-HT was positively correlated with serum calcium and with plasma PTHrP (r>0.37). Serum 5-HT was negatively correlated with milk fever incidence and with ketosis severity (most severe ketosis incidence recorded during the first 10 d postpartum; r<-0.33). Serum calcium and plasma glucose concentrations were negatively correlated with milk fever and ketosis severity, respectively (r<-0.39). These data indicate that 5-HT potentially plays a role in the regulation of calcium and glucose homeostasis during the transition period in cattle, which we previously demonstrated in rodents. Increased circulating concentrations of 5-HT might decrease milk fever at the onset of lactation and ketosis severity during the first 10 d postpartum in dairy cows. Understanding this physiological axis could help describe the

  5. Supplemental dietary protein for grazing dairy cows: effect on pasture intake and lactation performance.

    PubMed

    McCormick, M E; Ward, J D; Redfearn, D D; French, D D; Blouin, D C; Chapa, A M; Fernandez, J M

    2001-04-01

    One hundred twenty-four cows (92 multiparous and 32 primiparous) were used to evaluate the effect of grain supplements containing high crude protein [(22.8% CP, 5.3% rumen undegradable protein (RUP), dry matter basis], moderate CP (16.6% CP, 6.1% RUP), and moderate CP with supplemental RUP (16.2% CP, 10.8% RUP) on lactation performance of Holstein cows rotationally grazing annual ryegrass-oat pastures. Supplemental protein was provided by solvent extracted soybean meal in the high CP and moderate CP supplements and as a corn gluten meal-blood meal mixture (2.8:1) in the moderate CP, high RUP supplement. Cows were blocked according to previous mature milk equivalent production and calving date (partum group; 0 d in milk or postpartum group; 21 to 65 d in milk) and randomly assigned to dietary treatments. Grain was individually fed, at approximately a 1:3 grain to milk ratio, before a.m. and p.m milkings. The study was replicated during two grazing seasons that averaged 199 d. Cows had ad libitum access to bermudagrass hay while on pasture (dry matter intake = 1.3 kg/d). Protein supplementation had no effect on study long pasture dry matter (12.7 +/- 1.0 kg/d) or total dry matter (23.9 +/- 1.2 kg/d) consumption. Protein concentration did not affect actual milk yield of either calving group (high CP vs. moderate CP); however, postpartum group cows receiving high CP grain supplements maintained greater milk fat concentrations (3.34 vs. 3.11%), which led to higher fat-corrected milk (FCM) yields than control cows receiving moderate CP grain diets (30.3 vs. 28.9 kg/d). Crude protein concentration in milk of high CP-supplemented, postpartum group cows was also higher than moderate CP cows (3.42 vs. 3.27%). Additional RUP did not increase FCM yield above that generated by moderate CP grain diets for partum (34.3 vs. 32.9 kg/d) or postpartum-group cows (28.9 vs. 28.2 kg/d). Increasing CP concentration of grain supplement did not affect milk yield of Holstein cows grazing

  6. The effects of heat stress on protein metabolism in lactating Holstein cows.

    PubMed

    Gao, S T; Guo, J; Quan, S Y; Nan, X M; Fernandez, M V Sanz; Baumgard, L H; Bu, D P

    2017-04-05

    Heat stress (HS) decreases milk protein synthesis beyond what would be expected based on the concomitant reduction in feed intake. The aim of the present study was to evaluate the direct effects of HS on milk protein production. Four multiparous, lactating Holstein cows (101 ± 10 d in milk, 574 ± 36 kg of body weight, 38 ± 2 kg of milk/d) were individually housed in environmental chambers and randomly allocated to 1 of 2 groups in a crossover design. The study was divided into 2 periods with 2 identical experimental phases (control phase and trial phase) within each period. During phase 1 or control phase (9 d), all cows were housed in thermal neutral conditions (TN; 20°C, 55% humidity) and fed ad libitum. During phase 2 or treatment phase (9 d), group 1 was exposed to cyclical HS conditions (32 to 36°C, 40% humidity) and fed ad libitum, whereas group 2 remained in TN conditions but was pair-fed (PFTN) to their HS counterparts to eliminate the confounding effects of dissimilar feed intake. After a 30-d washout period in TN conditions, the study was repeated (period 2), inverting the environmental treatments of the groups relative to period 1: group 2 was exposed to HS and group 1 to PFTN conditions. Compared with PFTN conditions, HS decreased milk yield (17.0%), milk protein (4.1%), milk protein yield (19%), 4% fat-corrected milk (23%), and fat yield (19%). Apparent digestibility of dry matter, organic matter, neutral detergent fiber, acid detergent fiber, crude protein, and ether extract was increased (11.1-42.9%) in HS cows, as well as rumen liquor ammonia (before feeding 33.2%; after feeding 29.5%) and volatile fatty acid concentration (45.3%) before feeding. In addition, ruminal pH was reduced (9.5 and 6% before and after feeding, respectively) during HS. Heat stress decreased plasma free amino acids (AA; 17.1%) and tended to increase and increased blood, urine, and milk urea nitrogen (17.2, 243, and 24.5%, respectively). Further, HS cows had reduced

  7. Determination of protein and amino acid requirements of lactating sows using a population-based factorial approach.

    PubMed

    Strathe, A V; Strathe, A B; Theil, P K; Hansen, C F; Kebreab, E

    2015-08-01

    Determination of appropriate nutritional requirements is essential to optimize the productivity and longevity of lactating sows. The current recommendations for requirements do not consider the large variation between animals. Therefore, the aim of this study was to determine the amino acid recommendations for lactating sows using a stochastic modeling approach that integrates population variation and uncertainty of key parameters into establishing nutritional recommendations for lactating sows. The requirement for individual sows was calculated using a factorial approach by adding the requirement for maintenance and milk. The energy balance of the sows was either negative or zero depending on feed intake being a limiting factor. Some parameters in the model were sow-specific and others were population-specific, depending on state of knowledge. Each simulation was for 1000 sows repeated 100 times using Monte Carlo simulation techniques. BW, back fat thickness of the sow, litter size (LS), average litter gain (LG), dietary energy density and feed intake were inputs to the model. The model was tested using results from the literature, and the values were all within ±1 s.d. of the estimated requirements. Simulations were made for a group of low- (LS=10 (s.d.=1), LG=2 kg/day (s.d.=0.6)), medium- (LS=12 (s.d.=1), LG=2.5 kg/day (s.d.=0.6)) and high-producing (LS=14 (s.d.=1), LG=3.5 kg/day (s.d.=0.6)) sows, where the average requirement was the result. In another simulation, the requirements were estimated for each week of lactation. The results were given as the median and s.d. The average daily standardized ileal digestible (SID) protein and lysine requirements for low-, medium- and high-producing sows were 623 (CV=2.5%) and 45.1 (CV=4.8%); 765 (CV=4.9%) and 54.7 (CV=7.0%); and 996 (CV=8.5%) and 70.8 g/day (CV=9.6%), respectively. The SID protein and lysine requirements were lowest at week 1, intermediate at week 2 and 4 and the highest at week 3 of lactation. The

  8. [C-reactive protein and lactate dehydrogenase as single prognostic factors of severity in acute pancreatitis].

    PubMed

    Zrnić, Irena Krznarić; Milić, Sandra; Fisić, Elizabeta; Radić, Mladen; Stimac, Davor

    2007-01-01

    Ranson and Glasgow scores are routinely used for prediction of severity in acute pancreatitis. We undertook a prospective study to investigate the role of lactate dehydrogenase (LDH) and C-reactive protein (CRP) as potential single predictors of severity in acute pancreatitis. In our study we included 100 patients with diagnosis of acute pancreatitis admitted to our hospital during last two years. The inclusion criteria consisted of a combination of clinical features, a typical case history, elevation of serum pancreatic enzymes and diagnosis confirmed by imaging studies (ultrasound or computerised tomography). We used Ranson score for assesment of severity and compared it with single parameters as LDH and CRP on the first and the third day after admission. Cut off values for predicting local and systemic complications were > or =3 for Ranson score, 320 IU for LDH and 5 mg/L for CRP. Ranson score showed highest sensitivity in the prediction of local and systemic complication of acute pancreatitis. Specificity and diagnostic accuracy were highest for LDH on the first day (67.74; 57%). Diagnostic accuracy for Ranson score and CRP on the third day after admission was around 50%. We can conclude that LDH and CRP are available, simple and economical biochemical parameters that can help us predict complications of acute pancreatitis in the early phase of the disease. They showed similar diagnostic accuracy as the far more clinically used Ranson score.

  9. Evaluation of five lactation curve models fitted for fat:protein ratio of milk and daily energy balance.

    PubMed

    Buttchereit, N; Stamer, E; Junge, W; Thaller, G

    2010-04-01

    Selection for milk yield increases the metabolic load of dairy cows. The fat:protein ratio of milk (FPR) could serve as a measure of the energy balance status and might be used as a selection criterion to improve metabolic stability. The fit of different fixed and random regression models describing FPR and daily energy balance was tested to establish appropriate models for further genetic analyses. In addition, the relationship between both traits was evaluated for the best fitting model. Data were collected on a dairy research farm running a bull dam performance test. Energy balance was calculated using information on milk yield, feed intake per day, and live weight. Weekly FPR measurements were available. Three data sets were created containing records of 577 primiparous cows with observations from lactation d 11 to 180 as well as records of 613 primiparous cows and 96 multiparous cows with observations from lactation d 11 to 305. Five well-established parametric functions of days in milk (Ali and Schaeffer, Guo and Swalve, Wilmink, Legendre polynomials of third and fourth degree) were chosen for modeling the lactation curves. Evaluation of goodness of fit was based on the corrected Akaike information criterion, the Bayesian information criterion, correlation between the real observation and the estimated value, and on inspection of the residuals plotted against days in milk. The best model was chosen for estimation of correlations between both traits at different lactation stages. Random regression models were superior compared with the fixed regression models. In general, the Ali and Schaeffer function appeared most suitable for modeling both the fixed and the random regression part of the mixed model. The FPR is greatest in the initial lactation period when energy deficit is most pronounced. Energy balance stabilizes at the same point as the decrease in FPR stops. The inverted patterns indicate a causal relationship between the 2 traits. A common pattern was

  10. Assessment of the relationships among coagulopathy, hyperfibrinolysis, plasma lactate, and protein C in dogs with spontaneous hemoperitoneum.

    PubMed

    Fletcher, Daniel J; Rozanski, Elizabeth A; Brainard, Benjamin M; de Laforcade, Armelle M; Brooks, Marjory B

    2016-01-01

    To relate coagulation and fibrinolysis derangements to shock severity as reflected by plasma lactate concentrations in dogs with spontaneous hemoperitoneum (SHP) and determine the impact on transfusions. Prospective, observational, case-control study. Three veterinary teaching hospitals. Twenty-eight client-owned dogs with SHP and 28 breed- and age-matched control dogs. None. Blood samples for platelet counts, coagulation, and anticoagulant assays (prothrombin time, activated partial thromboplastin time, fibrinogen, antithrombin, and protein C, thromboelastography [TEG]), fibrinolysis testing (d-dimer and TEG lysis parameters with and without the addition of 50 U/mL of tissue plasminogen activator [TEG LY30 measured with the addition of 50 U/mL of tPA to the blood sample, LY3050 and TEG LY60 measured with the addition of 50 U/mL of tPA to the blood sample, LY6050 ; LY30 and LY60]), and plasma lactate as an indicator of severity of shock were collected from SHP dogs at the time of diagnosis. SHP dogs were hypocoagulable (prolonged prothrombin time and activated partial thromboplastin time, decreased TEG maximum amplitude) and hyperfibrinolytic (increased LY3050 and TEG LY6050 ) compared to controls. The severity of hypocoagulability was related to protein C activity, while the severity of hyperfibrinolysis was related to plasma lactate concentration. Among the 18 dogs discharged from the hospital, LY3050 was significantly associated with the dose of fresh frozen plasma administered, but none of the parameters were associated with the dose of red blood cells administered. Dogs with SHP have evidence of hypocoagulability, protein C deficiency, and hyperfibrinolysis. Parameters of hyperfibrinolysis were related to plasma lactate concentrations and volume of plasma transfused during hospitalization. These derangements resemble those found in people with acute coagulopathy of trauma and shock, and activation of protein C may be a common feature to both syndromes.

  11. Effects of PTHrP on chondrocytes of sika deer antler.

    PubMed

    Guo, Bin; Wang, Shou-Tang; Duan, Cui-Cui; Li, Dang-Dang; Tian, Xue-Chao; Wang, Qu-Yuan; Yue, Zhan-Peng

    2013-11-01

    Parathyroid-hormone-related peptide (PTHrP) is an important regulator of chondrocyte differentiation in growth plates but little is known about its role in deer antler cartilage. The aim of the present study was to use the deer antler as a model to determine the possible role of PTHrP in regulating chondrocyte differentiation of deer antler. PTHrP and its receptor PTH1R mRNA were highly expressed in the perichondrium and cartilage of sika deer antler, as shown by in situ hybridization. Chondrocytes of deer antler were identified by toluidine blue staining of glycosaminoglycan and immunocytochemical staining of type II collagen (Col II). Treatment with PTHrP (1-34) reduced the expression of prehypertrophic chondrocyte marker Col IX and hypertrophic chondrocyte marker Col X. In order to confirm the mechanism of action of PTHrP, we initially examined the expression of cyclin D1, Bcl-2 and runt-related transcription factor 2 (Runx2) in sika deer antler by in situ hybridization and found that cyclin D1, Runx2 and Bcl-2 mRNA were also expressed in antler chondrocytes. Exogenous PTHrP induced the expression of cyclin D1 and Bcl-2 mRNA by various signalling pathways, whereas it inhibited Runx2 expression through PKA, p38MAPK, MEK and PI3K signalling pathways. Thus, PTHrP might promote the proliferation of antler chondrocytes and prevent their differentiation; it might furthermore influence the growth and development of sika deer antler.

  12. Lactation Consultant

    MedlinePlus

    ... human lactation. Job description Lactation consultants educate women, families, health professionals, and the community about breast feeding and human lactation; facilitate the development of policies which protect, promote, and support breastfeeding; ...

  13. Genetic associations between milk fat-to-protein ratio, milk production and fertility in the first two lactations of Thai Holsteins dairy cattle.

    PubMed

    Puangdee, Somsook; Duangjinda, Monchai; Boonkum, Wuttigrai; Katawatin, Suporn; Buaban, Sayan; Thepparat, Mongkol

    2017-05-01

    The aims of this study were to estimate, simultaneously, the genetic parameters of test-day milk fat-to-protein ratio (FPR), test-day milk yield (MY), and days-open (DO) in the first two lactations of Thai Holsteins. A total of 76 194 test-day production records collected from 8874 cows with 8674 DO records between 2001 and 2011 from different lactations were treated as separated traits. The estimates of heritability for test-day FPR in the first lactation showed an increasing trend, whereas the estimates in the second lactation showed a U-shape trend. Genetic correlations for FPR-DO and MY-DO showed a decreasing trend along days in milk (DIM) in both lactations, whereas genetic correlations for FPR-MY increased along DIM in the first lactation but decreased in the second lactation. Genetic correlations of FPR between consecutive DIM were moderate to high, which showed the effectiveness of simultaneous analyses. Selection of FPR in the early stage has no adverse effect on MY and DO for the first lactation but has a negative effect on MY and positive effect on DO for the second lactation. This study showed that genetic improvement of the energy balance using FPR, MY and DO with multi-trait test day model could be applied in a Thailand dairy cattle breeding program. © 2016 Japanese Society of Animal Science.

  14. Closed loop control of lactate concentration in mammalian cell culture by Raman spectroscopy leads to improved cell density, viability, and biopharmaceutical protein production.

    PubMed

    Matthews, Thomas E; Berry, Brandon N; Smelko, John; Moretto, Justin; Moore, Brandon; Wiltberger, Kelly

    2016-11-01

    Accumulation of lactate in mammalian cell culture often negatively impacts culture performance, impeding production of therapeutic proteins. Many efforts have been made to limit the accumulation of lactate in cell culture. Here, we describe a closed loop control scheme based on online spectroscopic measurements of glucose and lactate concentrations. A Raman spectroscopy probe was used to monitor a fed-batch mammalian cell culture and predict glucose and lactate concentrations via multivariate calibration using partial least squares regression (PLS). The PLS models had a root mean squared error of prediction (RMSEP) of 0.27 g/L for glucose and 0.20 g/L for lactate. All glucose feeding was controlled by the Raman PLS model predictions. Glucose was automatically fed when lactate levels were beneath a setpoint (either 4.0 or 2.5 g/L) and glucose was below its own setpoint (0.5 g/L). This control scheme was successful in maintaining lactate levels at an arbitrary setpoint throughout the culture, as compared to the eventual accumulate of lactate to 8.0 g/L in the historical process. Automated control of lactate by restricted glucose feeding led to improvements in culture duration, viability, productivity, and robustness. Culture duration was extended from 11 to 13 days, and harvest titer increased 85% over the historical process. Biotechnol. Bioeng. 2016;113: 2416-2424. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  15. Plasma lactate, GH and GH-binding protein levels in exercise following BCAA supplementation in athletes.

    PubMed

    De Palo, E F; Gatti, R; Cappellin, E; Schiraldi, C; De Palo, C B; Spinella, P

    2001-01-01

    Branched chain amino acids (BCAA) stimulate protein synthesis, and growth hormone (GH) is a mediator in this process. A pre-exercise BCAA ingestion increases muscle BCAA uptake and use. Therefore after one month of chronic BCAA treatment (0.2 gkg(-1) of body weight), the effects of a pre-exercise oral supplementation of BCAA (9.64 g) on the plasma lactate (La) were examined in triathletes, before and after 60 min of physical exercise (75% of VO2 max). The plasma levels of GH (pGH) and of growth hormone binding protein (pGHBP) were also studied. The end-exercise La of each athlete was higher than basal. Furthermore, after the chronic BCAA treatment, these end-exercise levels were lower than before this treatment (8.6+/-0.8 mmol L(-1) after vs 12.8+/-1.0 mmol L(-1) before treatment; p < 0.05 [mean +/- std. err.]). The end-exercise pGH of each athlete was higher than basal (p < 0.05). Furthermore, after the chronic treatment, this end-exercise pGH was higher (but not significantly, p = 0.08) than before this treatment (12.2+/-2.0 ng mL(-1) before vs 33.8+/-13.6 ngmL(-1) after treatment). The end-exercise pGHBP was higher than basal (p < 0.05); and after the BCAA chronic treatment, this end-exercise pGHBP was 738+/-85 pmol L(-1) before vs 1691+/-555 pmol L(-1) after. pGH/pGHBP ratio was unchanged in each athlete and between the groups, but a tendency to increase was observed at end-exercise. The lower La at the end of an intense muscular exercise may reflect an improvement of BCAA use, due to the BCAA chronic treatment. The chronic BCAA effects on pGH and pGHBP might suggest an improvement of muscle activity through protein synthesis.

  16. Effect of a high-protein diet on food intake and liver metabolism during pregnancy, lactation and after weaning in mice.

    PubMed

    Kuhla, Björn; Kucia, Marzena; Görs, Solvig; Albrecht, Dirk; Langhammer, Martina; Kuhla, Siegfried; Metges, Cornelia C

    2010-07-01

    Major hepatic metabolic pathways are involved in the control of food intake but how dietary proteins affect global metabolism to adjust food intake is incompletely understood, particularly under physiological challenging conditions such as lactation. In order to identify these molecular events, mice were fed a high-protein (HP) diet from pregnancy, during lactation until after weaning and compared with control fed counterparts. Liver specimens were analyzed for regulated proteins using 2-DE and MALDI-TOF-MS and plasma samples for metabolites. Based on the 26 differentially expressed proteins associated with depleted liver glycogen content, elevated urea and citrulline plasma concentrations, we conclude that HP feeding during lactation leads to an activated amino acid, carbohydrate and fatty acid catabolism while it activates gluconeogenesis. From pregnancy to lactation, plasma arginine, tryptophan, serine, glutamine and cysteine decreased, whereas urea concentrations increased in both groups. Concomitantly, hepatic glycogen content decreased while total fat content remained unaltered in both groups. Consideration of 59 proteins differentially expressed between pregnancy and lactation highlights different strategies of HP and control fed mice to meet energy requirements for lactation by adjusting amino acid degradation, carbohydrate and fat metabolism, citrate cycle, but also ATP-turnover, protein folding, secretion of proteins and (de)activation of transcription factors.

  17. Genetic parameters for lactation traits of milking ewes: protein content and composition, fat, somatic cells and individual laboratory cheese yield

    PubMed Central

    Othmane, Med Houcine; Carriedo, Juan Antonio; San Primitivo, Fermin; De la Fuente, Luis Fernando

    2002-01-01

    The effects of some environmental variation factors and the genetic parameters for total milk traits (fat content, protein content, casein content, serum protein content, lactation mean of individual laboratory cheese yield (LILCY), lactation mean of somatic cell count (LSCC), and milk yield) were estimated from the records of 1 111 Churra ewes. Genetic parameters were estimated by multivariate REML. Heritability for fat content was low (0.10) as is usually found in the Churra breed. Heritabilities for protein content, casein content, serum protein content, LILCY, milk yield and somatic cell count were 0.31, 0.30, 0.22, 0.09, 0.26 and 0.11, respectively. The highest heritability estimates were for protein and casein contents. Casein content is not advisable as an alternative to protein content as a selection criterion for cheese yield improvement; it does not have any compelling advantages and its measurement is costly. Our results for LSCC indicated that efforts should focus on improving the level of management rather than selecting for somatic cells, in the actual conditions of the Churra breed. PMID:12427387

  18. The dietary protein/carbohydrate ratio differentially modifies lipogenesis and protein synthesis in the mammary gland, liver and adipose tissue during gestation and lactation.

    PubMed

    Velázquez-Villegas, Laura A; Tovar, Armando R; López-Barradas, Adriana M; Torres, Nimbe

    2013-01-01

    During gestation and lactation, a series of metabolic changes that are affected by the diet occurs in various organs of the mother. However, little is known about how the dietary protein (DP)/carbohydrate (DCH) ratio regulates the expression of metabolic genes in the mother. Therefore, the purpose of this work was to study the effect of consuming different percentages of DP/DCH, specifically 10/73, 20/63 and 30/53%, on the expression of genes involved in lipogenesis and protein synthesis in the mammary gland, liver and adipose tissue during gestation and lactation in dams. While the amount of weight gained during gestation was similar for all groups, only dams fed with 30/53% DP/DCH maintained their weight during lactation. In the mammary gland, the expression of the genes involved in lipogenesis, specifically SREBP1 and FAS, was dramatically increased, and the expression of the genes involved in protein synthesis, such as mTOR1, and the phosphorylation of its target protein, S6K, were also increased throughout pregnancy and lactation, regardless of the concentration of DP/DCH. In the liver and adipose tissue, the expression of the genes and proteins involved in lipid metabolism was dependent on the proportion of DP/DCH. The consumption of a low-protein/high-carbohydrate diet increased the expression of lipogenic genes in the liver and adipose tissue and the amount of lipid deposition in the liver. Conversely, the consumption of a high-protein/low-carbohydrate diet increased the expression of genes involved in amino acid oxidation in the liver during gestation. The metabolic adaptations reflected by the changes in the expression of metabolic genes indicate that the mammary gland has a priority for milk synthesis, whereas the adaptations in the liver and adipose tissue are responsible for providing nutrients to the mammary gland to sustain milk synthesis.

  19. Expression of PTHrP and PTH/PTHrP receptor 1 in the superior cervical ganglia of rats.

    PubMed

    Filipović, Natalija; Vrdoljak, Marija; Vuica, Ana; Jerić, Milka; Jeličić Kadić, Antonia; Utrobičić, Toni; Mašek, Tomislav; Grković, Ivica

    2014-12-01

    PTHrP and its receptor PTHR1 are found in the CNS and peripheral nervous system. The presence of PTHrP mRNA has been detected in the superior cervical ganglion (SCG), but there are no data on the cellular distribution of PTHrP and PTHR1 in the SCG. Although it is known that ovarian activity and reproductive status influence sympathetic activity, and the PTHrP/PTHR1 system is influenced by estrogens in different tissues, it is not known whether these factors have a similar effect on expression of PTHrP and PTHR1 in the nervous system. Hence, we investigated the presence and distribution of PTHrP and PTHR1 in neurons and glia of the SCG of rats, as well as the influence of ovariectomy on their expression, by using immunohistochemistry. PTHrP and PTHR1 immunoreactivity was observed in cytoplasm as well as in nuclei of almost all neurons in the SCG. In male rats, intensity of PTHrP fluorescence was significantly higher in cytoplasm of NPY-, in comparison to NPY+ neurons (p < 0.05). In female rats, 2 months post-ovariectomy, significantly lower intensity of PTHrP fluorescence in cytoplasm of the SCG neurons was observed in comparison to sham operated animals (p < 0.05). In addition to neurons, PTHrP and PTHR1 immunoreactivity was observed in most of the glia and was not influenced by ovariectomy. Results show the expression of PTHrP and its receptor, PTHR1, in the majority of neurons and glial cells in the SCG of rats. Expression of PTHrP, but not PTHR1 in the cytoplasm of SCG neurons is influenced by ovarian activity.

  20. Major whey proteins in donkey's milk: effect of season and lactation stage. Implications for potential dietary interventions in human diseases.

    PubMed

    D'Alessandro, Angela G; Martemucci, Giovanni; Jirillo, Emilio; De Leo, Vincenzo

    2011-06-01

    According to current literature, donkey's milk has been suggested as a hypoallergenic substitute in children affected by cow's milk protein allergy as well as a promising nutraceutical for aged people. However, the biologically active components of donkey's milk have not yet completely elucidated. In this framework this study is aimed at measuring α-lactalbumin (α-LA), β-lactoglobulin (β-LG), and lysozyme (LYS), the principal whey proteins in donkey's milk, in relation to lactation stage and production season. Analysis were performed by reversed-phase high-performance liquid chromatography. α-LA, β-LG, and LYS resulted to be affected by lactation stage (P < 0.01) and production season (P < 0.01). Overall, the protein content was higher (0.01 > P < 0.05) during the first four lactation's months and decreased until the month 8. The β-LG was the major protein (1.75 mg mL(-1) as mean; peak 2.24 ± 0.09 mg mL(-1)), while the α-LA had a mean concentration of 1.32 mg mL(-1) and peaked at month 1 (1.57 ± 0.09 mg mL(-1)) and LYS (0.66 mg mL(-1) as mean) showed the highest value equal to 0.76 ± 0.03 mg mL(-1). The highest (P < 0.01) concentration of all proteins was recorded at spring (α-LA: 1.69 mL(-1); β-LG: 2.07 mL(-1); LYS: 0.76 mL(-1)).

  1. Effects of L-carnitine supplementation on milk production, litter gains and back-fat thickness in sows with a low energy and protein intake during lactation.

    PubMed

    Ramanau, A; Kluge, H; Eder, K

    2005-05-01

    The present study investigated the effect of L-carnitine supplementation during pregnancy (125 mg/d) and lactation (250 mg/d) on milk production, litter gains and back-fat thickness in sows fed a low-energy and low-protein diet during lactation. Sows supplemented with L-carnitine produced more milk on days 11 and 18 of lactation (+18 %; P<0.05) and had higher litter gains during suckling (+20 %; P<0.01) than control sows. Loss of body weight during lactation was similar in both groups, but sows supplemented with L-carnitine had a greater reduction of back-fat thickness (+45 %; P<0.05) during lactation than control sows. In conclusion, this study shows that L-carnitine increases milk production and litter gains in sows in a strongly negative energy and N balance, and enhances body fat mobilisation.

  2. Parathyroid hormone-related protein and its receptors: nuclear functions and roles in the renal and cardiovascular systems, the placental trophoblasts and the pancreatic islets

    PubMed Central

    Clemens, Thomas L; Cormier, Sarah; Eichinger, Anne; Endlich, Karlhans; Fiaschi-Taesch, Nathalie; Fischer, Evelyne; Friedman, Peter A; Karaplis, Andrew C; Massfelder, Thierry; Rossert, Jérôme; Schlüter, Klaus-Dieter; Silve, Caroline; Stewart, Andrew F; Takane, Karen; Helwig, Jean-Jacques

    2001-01-01

    The cloning of the so-called ‘parathyroid hormone-related protein' (PTHrP) in 1987 was the result of a long quest for the factor which, by mimicking the actions of PTH in bone and kidney, is responsible for the hypercalcemic paraneoplastic syndrome, humoral calcemia of malignancy. PTHrP is distinct from PTH in a number of ways. First, PTHrP is the product of a separate gene. Second, with the exception of a short N-terminal region, the structure of PTHrP is not closely related to that of PTH. Third, in contrast to PTH, PTHrP is a paracrine factor expressed throughout the body. Finally, most of the functions of PTHrP have nothing in common with those of PTH. PTHrP is a poly-hormone which comprises a family of distinct peptide hormones arising from post-translational endoproteolytic cleavage of the initial PTHrP translation products. Mature N-terminal, mid-region and C-terminal secretory forms of PTHrP are thus generated, each of them having their own physiologic functions and probably their own receptors. The type 1 PTHrP receptor, binding both PTH(1-34) and PTHrP(1-36), is the only cloned receptor so far. PTHrP is a PTH-like calciotropic hormone, a myorelaxant, a growth factor and a developmental regulatory molecule. The present review reports recent aspects of PTHrP pharmacology and physiology, including: (a) the identification of new peptides and receptors of the PTH/PTHrP system; (b) the recently discovered nuclear functions of PTHrP and the role of PTHrP as an intracrine regulator of cell growth and cell death; (c) the physiological and developmental actions of PTHrP in the cardiovascular and the renal glomerulo-vascular systems; (d) the role of PTHrP as a regulator of pancreatic beta cell growth and functions, and, (e) the interactions of PTHrP and calcium-sensing receptors for the control of the growth of placental trophoblasts. These new advances have contributed to a better understanding of the pathophysiological role of PTHrP, and will help to identify

  3. Effects of PTHrP on expression of MMP9 and MMP13 in sika deer antler chondrocytes.

    PubMed

    Wang, Shou-Tang; Gao, Ying-Jie; Duan, Cui-Cui; Li, Dang-Dang; Tian, Xue-Chao; Zhang, Qiao-Ling; Guo, Bin; Yue, Zhan-Peng

    2013-12-01

    Deer antlers are the only mammalian appendages to display an annual cycle of full regeneration. However, little is known about the molecular mechanisms of antler regeneration. Our previous study has demonstrated that parathyroid hormone-related peptide (PTHrP) can promote proliferation of antler chondrocytes and inhibit its differentiation, but the mechanism underlying such regulation is not fully understood. We have determined the role of PTHrP on the mRNA expression of matrix metalloproteinase-9 (MMP9) and MMP13 in the antler chondrocytes. The possible pathways that transduce PTHrP effects were examined. In situ hybridization showed that MMP9 and MMP13 were mainly localized in the dermal fibroblasts, perichondrium, and cartilage in the sika deer antler, of which MMP9 and MMP13 were highly expressed in the chondrocytes. Exogenous PTHrP could inhibit the expression of MMP9 and MMP13 in the antler chondrocytes. The inhibitory effect of PTHrP on MMP9 was abolished by JNK inhibitor, SP600125, while P38MAPK inhibitor SB203850 and PKC inhibitor GF109203X could rescue the inhibitory effect of PTHrP on MMP13. The results suggest that PTHrP can inhibit MMP9 expression by JNK signaling pathway and MMP13 expression by p38MAPK and PKC signaling pathways in the antler chondrocytes. Thus PTHrP is involved in the control of antler chondrocytes maturation and cartilage matrix degradation. © 2013 International Federation for Cell Biology.

  4. Maternal protein restriction in the rat during pregnancy and/or lactation alters cognitive and anxiety behaviors of female offspring.

    PubMed

    Reyes-Castro, L A; Rodriguez, J S; Charco, R; Bautista, C J; Larrea, F; Nathanielsz, P W; Zambrano, E

    2012-02-01

    Maternal protein deficiencies can developmentally program offspring to lifelong dysfunction of many physiological systems. We hypothesized that maternal isocaloric low protein diet during fetal and early postnatal development would negatively affect female offspring anxiety, exploration, associative learning and motivation as measured by the elevated plus maze (EPM), open field test (OFT), operant conditioning and the progressive ratio task, respectively. Control mothers (C) received a 20% casein diet and restricted mothers (R) a 10% casein diet to provide four groups: CC, RR, CR, and RC (first letter pregnancy diet and second lactation diet) to enable evaluation of offspring effects influenced by maternal diet during pregnancy and lactation. Maternal protein restriction decreased open arm time and distance in RR and RC offspring, increased anxiety behavior, in the EPM. In the OFT, the RR and RC offspring displayed decreased exploration (increased stress) as indexed by decreased distance in the center zone. These behaviors in the EPM and OFT was associated with increased corticosterone levels during an immobilization test in the RR offspring with intermediary effects in the RC offspring. Learning impairment was observed in the RR, CR and RC offspring during fixed ratio 5 schedule of reinforcement. Motivational effects were measured in RR offspring responding less, decreased motivation, and CR offspring making more responses, increased motivation, than CC offspring. These findings reveal the negative effects of developmental protein restriction on female offspring behavior. The underlying basis for these negative outcomes remains to be elucidated. Copyright © 2011 ISDN. Published by Elsevier Ltd. All rights reserved.

  5. 1,25-dihydroxyvitamin D and PTHrP mediated malignant hypercalcemia in a seminoma.

    PubMed

    Rodríguez-Gutiérrez, René; Zapata-Rivera, Maria Azucena; Quintanilla-Flores, Dania Lizeth; Camara-Lemarroy, Carlos Rodrigo; Lavalle-Gonzalez, Fernando Javier; González-González, José Gerardo; Villarreal-Pérez, Jesús Zacarías

    2014-04-10

    Seminomas have been rarely associated with malignant hypercalcemia. The responsible mechanism of hypercalcemia in this setting has been described to be secondary to 1,25-dihydroxyvitamin D secretion. The relationship with PTHrP has not been determined or studied.The aim of this study is to describe and discuss the case and the pathophysiological mechanisms involved in a malignant hypercalcemia mediated by 1,25-dihydroxyvitamin D and PTHrP cosecretion in a patient with seminoma. A 35-year-old man was consulted for assessment and management of severe hypercalcemia related to an abdominal mass. Nausea, polyuria, polydipsia, lethargy and confusion led him to the emergency department. An abdominal and pelvic enhanced CT confirmed a calcified pelvic mass, along with multiple retroperitoneal lymphadenopathy. Chest x-ray revealed "cannon ball" pulmonary metastases. The histopathology result was consistent with a seminoma. Serum calcium was 14.7 mg/dl, PTH was undetectable, 25-dihydroxyvitamin D was within normal values and PTHrP and 1,25-dihydroxyvitamin were elevated (35.0 pg/ml, and 212 pg/ml, respectively). After the first cycle of chemotherapy with bleomycin, etoposide and cisplatin, normocalcemia was restored. Both PTHrP and 1,25-dihydroxyvitamin D, dropped dramatically to 9.0 pg/ml and 8.0 pg/ml, respectively. The association of seminoma and malignant hypercalcemia is extremely rare. We describe a case of a patient with a seminoma and malignant hypercalcemia related to paraneoplastic cosecretion of 1,25-dihydroxyvitamin D and PTHrP. After successful chemotherapy, calcium, PTHrP and 1,25-Dihydroxyvitamin D returned to normal values.

  6. 1,25-dihydroxyvitamin D and PTHrP mediated malignant hypercalcemia in a seminoma

    PubMed Central

    2014-01-01

    Background Seminomas have been rarely associated with malignant hypercalcemia. The responsible mechanism of hypercalcemia in this setting has been described to be secondary to 1,25-dihydroxyvitamin D secretion. The relationship with PTHrP has not been determined or studied. The aim of this study is to describe and discuss the case and the pathophysiological mechanisms involved in a malignant hypercalcemia mediated by 1,25-dihydroxyvitamin D and PTHrP cosecretion in a patient with seminoma. Case presentation A 35-year-old man was consulted for assessment and management of severe hypercalcemia related to an abdominal mass. Nausea, polyuria, polydipsia, lethargy and confusion led him to the emergency department. An abdominal and pelvic enhanced CT confirmed a calcified pelvic mass, along with multiple retroperitoneal lymphadenopathy. Chest x-ray revealed “cannon ball” pulmonary metastases. The histopathology result was consistent with a seminoma. Serum calcium was 14.7 mg/dl, PTH was undetectable, 25-dihydroxyvitamin D was within normal values and PTHrP and 1,25-dihydroxyvitamin were elevated (35.0 pg/ml, and 212 pg/ml, respectively). After the first cycle of chemotherapy with bleomycin, etoposide and cisplatin, normocalcemia was restored. Both PTHrP and 1,25-dihydroxyvitamin D, dropped dramatically to 9.0 pg/ml and 8.0 pg/ml, respectively. Conclusion The association of seminoma and malignant hypercalcemia is extremely rare. We describe a case of a patient with a seminoma and malignant hypercalcemia related to paraneoplastic cosecretion of 1,25-dihydroxyvitamin D and PTHrP. After successful chemotherapy, calcium, PTHrP and 1,25-Dihydroxyvitamin D returned to normal values. PMID:24721620

  7. Roasted soybeans, blood meal, and tallow as sources of fat and ruminally undegradable protein in the diets of lactating cows.

    PubMed

    Pires, A V; Eastridge, M L; Firkins, J L

    1996-09-01

    The effect of roasted soybeans, blood meal, and tallow as sources of fat and RUP for lactating dairy cows was studied. Forty-five cows, blocked by age, calving date, and milk yield during the previous lactation, were assigned randomly to the following treatments (ingredient in the DM, RUP as a percentage of CP, and fat in the DM, respectively): 1) soybean meal (16, 35, and 3.2%), 2) whole roasted soybeans (18, 40, and 6.2%), 3) ground roasted soybeans (18, 40, and 6.2%), 4) blood meal (2.7, 40, and 3.2%), and 5) blood meal plus tallow (2.7 and 3, 40, and 6.2%). Diets were fed from wk 3 to 18 of lactation and consisted of 20% alfalfa silage, 30% corn silage, and 50% concentrate. The DMI of blood meal and whole roasted soybean diets was about 11% lower than DMI of the soybean meal diet. Milk yield (38.4 kg/d) and milk fat percentage (3.37%) were similar among diets. The roasted soybean diets resulted in the lowest milk protein percentage. Less than 2.7% blood meal might be advisable for diets fed to high yielding dairy cows to avoid reduced DMI.

  8. The Wnt/β-Catenin Pathway Interacts Differentially with PTHrP Signaling to Control Chondrocyte Hypertrophy and Final Maturation

    PubMed Central

    Guo, Xizhi; Mak, Kinglun Kingston; Taketo, Makoto M.; Yang, Yingzi

    2009-01-01

    Sequential proliferation, hypertrophy and maturation of chondrocytes are required for proper endochondral bone development and tightly regulated by cell signaling. The canonical Wnt signaling pathway acts through β-catenin to promote chondrocyte hypertrophy whereas PTHrP signaling inhibits it by holding chondrocytes in proliferating states. Here we show by genetic approaches that chondrocyte hypertrophy and final maturation are two distinct developmental processes that are differentially regulated by Wnt/β-catenin and PTHrP signaling. Wnt/β-catenin signaling regulates initiation of chondrocyte hypertrophy by inhibiting PTHrP signaling activity, but it does not regulate PTHrP expression. In addition, Wnt/β-catenin signaling regulates chondrocyte hypertrophy in a non-cell autonomous manner and Gdf5/Bmp signaling may be one of the downstream pathways. Furthermore, Wnt/β-catenin signaling also controls final maturation of hypertrophic chondrocytes, but such regulation is PTHrP signaling-independent. PMID:19557172

  9. The Wnt/beta-catenin pathway interacts differentially with PTHrP signaling to control chondrocyte hypertrophy and final maturation.

    PubMed

    Guo, Xizhi; Mak, Kinglun Kingston; Taketo, Makoto M; Yang, Yingzi

    2009-06-26

    Sequential proliferation, hypertrophy and maturation of chondrocytes are required for proper endochondral bone development and tightly regulated by cell signaling. The canonical Wnt signaling pathway acts through beta-catenin to promote chondrocyte hypertrophy whereas PTHrP signaling inhibits it by holding chondrocytes in proliferating states. Here we show by genetic approaches that chondrocyte hypertrophy and final maturation are two distinct developmental processes that are differentially regulated by Wnt/beta-catenin and PTHrP signaling. Wnt/beta-catenin signaling regulates initiation of chondrocyte hypertrophy by inhibiting PTHrP signaling activity, but it does not regulate PTHrP expression. In addition, Wnt/beta-catenin signaling regulates chondrocyte hypertrophy in a non-cell autonomous manner and Gdf5/Bmp signaling may be one of the downstream pathways. Furthermore, Wnt/beta-catenin signaling also controls final maturation of hypertrophic chondrocytes, but such regulation is PTHrP signaling-independent.

  10. Serotonin Regulates Calcium Homeostasis in Lactation by Epigenetic Activation of Hedgehog Signaling

    PubMed Central

    Laporta, Jimena; Keil, Kimberly P.; Weaver, Samantha R.; Cronick, Callyssa M.; Prichard, Austin P.; Crenshaw, Thomas D.; Heyne, Galen W.; Vezina, Chad M.; Lipinski, Robert J.

    2014-01-01

    Calcium homeostasis during lactation is critical for maternal and neonatal health. We previously showed that nonneuronal/peripheral serotonin [5-hydroxytryptamine (5-HT)] causes the lactating mammary gland to synthesize and secrete PTHrP in an acute fashion. Here, using a mouse model, we found that genetic inactivation of tryptophan hydroxylase 1 (Tph1), which catalyzes the rate-limiting step in peripheral 5-HT synthesis, reduced circulating and mammary PTHrP expression, osteoclast activity, and maternal circulating calcium concentrations during the transition from pregnancy to lactation. Tph1 inactivation also reduced sonic hedgehog signaling in the mammary gland during lactation. Each of these deficiencies was rescued by daily injections of 5-hydroxy-L-tryptophan (an immediate precursor of 5-HT) to Tph1-deficient dams. We used immortalized mouse embryonic fibroblasts to demonstrate that 5-HT induces PTHrP through a sonic hedgehog-dependent signal transduction mechanism. We also found that 5-HT altered DNA methylation of the Shh gene locus, leading to transcriptional initiation at an alternate start site and formation of a variant transcript in mouse embryonic fibroblasts in vitro and in mammary tissue in vivo. These results support a new paradigm of 5-HT-mediated Shh regulation involving DNA methylation remodeling and promoter switching. In addition to having immediate implications for lactation biology, identification and characterization of a novel functional regulatory relationship between nonneuronal 5-HT, hedgehog signaling, and PTHrP offers new avenues for the study of these important factors in development and disease. PMID:25192038

  11. Decreased liver triglyceride content in adult rats exposed to protein restriction during gestation and lactation: role of hepatic triglyceride utilization

    PubMed Central

    Qasem, Rani J.; Li, Jing; Tang, Hee Man; Browne, Veron; Mendez, Claudia; Yablonski, Elizabeth; Pontiggia, Laura; D’mello, Anil P.

    2015-01-01

    We have previously demonstrated that protein restriction throughout gestation and lactation reduced liver triglyceride content in adult rat offspring. The mechanism(s) mediating the decrease in liver triglyceride content are not understood. The objective of the current study was to use a new group of pregnant animals and their offspring and determine the contribution of increased triglyceride utilization via the hepatic fatty acid oxidation and triglyceride secretory pathways to the reduction in liver triglyceride content. Pregnant Sprague-Dawley rats received either a control or a low protein diet throughout pregnancy and lactation. Pups were weaned onto laboratory chow on day 28 and sacrificed on day 65. Liver triglyceride content was reduced in male, but not female, low protein offspring both in the fed and fasted states. The reduction was accompanied by a trend towards higher liver carnitine palmitoyltransferase-1a activity suggesting increased fatty acid transport into the mitochondrial matrix. However, medium chain acyl CoA dehydrogenase activity within the mitochondrial matrix, expression of nuclear peroxisome proliferator activated receptor-α, and plasma levels of β-hydroxybutyrate were similar between low protein and control offspring indicating a lack of change in fatty acid oxidation. Hepatic triglyceride secretion, assessed by blocking peripheral triglyceride utilization and measuring serum triglyceride accumulation rate, and the activity of microsomal transfer protein were similar between low protein and control offspring. Since enhanced triglyceride utilization is not a significant contributor, the decrease in liver triglyceride content in male low protein offspring is likely due to alterations in liver fatty acid transport or triglyceride biosynthesis. PMID:25641378

  12. Milk yield and composition of lactating cows fed steam-flaked sorghum and graded concentrations of ruminally degradable protein.

    PubMed

    Santos, F A; Huber, J T; Theurer, C B; Swingle, R S; Simas, J M; Chen, K H; Yu, P

    1998-01-01

    To determine the effect of various amounts of ruminally undegradable protein in the diets of lactating cows fed steam-flaked sorghum, 24 Holstein cows (90 +/- 50 d in milk) were assigned to three treatments: 0.8% urea, 6% soybean meal, or 5% fish meal. Respective percentages of ruminally undegradable protein in the diets (as a percentage of crude protein) were 30, 35, and 39%. All diets contained 37% alfalfa hay; 3 to 5% cottonseed hulls; 10 to 13% whole cottonseed; 39% steam-flaked sorghum (360 g/L); 5% of a molasses, mineral, and vitamin supplement; and the different protein supplements. Intake of dry matter was higher for cows fed urea than for cows fed soybean meal or fish meal diets. In cows that yielded more than 40 kg/d of milk (4 cows per treatment), the soybean meal and fish meal diets resulted in higher yields of milk and 3.5% fat-corrected milk and a greater efficiency of conversion of feed to milk than did the urea diet. Cows that yielded less than 40 kg/d of milk (4 cows per treatment) at the beginning of treatment tended to yield more milk when fed urea than when fed the protein supplements. Nutrient digestibilities were not greatly affected by source of N, suggesting a beneficial effect of urea supplementation on nutrient digestibilities because replacement of protein supplements with cottonseed products caused the neutral detergent fiber content of the urea diet to be about 7% higher than that of the other diets. These data show that response to ruminally undegradable protein in diets of lactating cows fed steam-flaked sorghum was related to milk yield.

  13. Protein Conformational Landscapes and Catalysis. Influence of Active Site Conformations in the Reaction Catalyzed by L-Lactate Dehydrogenase

    PubMed Central

    Świderek, Katarzyna; Tuñón, Iñaki; Martí, Sergio; Moliner, Vicent

    2015-01-01

    In the last decade L-Lactate Dehydrogenase (LDH) has become an extremely useful marker in both clinical diagnosis and in monitoring the course of many human diseases. It has been assumed from the 80s that the full catalytic process of LDH starts with the binding of the cofactor and the substrate followed by the enclosure of the active site by a mobile loop of the protein before the reaction to take place. In this paper we show that the chemical step of the LDH catalyzed reaction can proceed within the open loop conformation, and the different reactivity of the different protein conformations would be in agreement with the broad range of rate constants measured in single molecule spectrometry studies. Starting from a recently solved X-ray diffraction structure that presented an open loop conformation in two of the four chains of the tetramer, QM/MM free energy surfaces have been obtained at different levels of theory. Depending on the level of theory used to describe the electronic structure, the free energy barrier for the transformation of pyruvate into lactate with the open conformation of the protein varies between 12.9 and 16.3 kcal/mol, after quantizing the vibrations and adding the contributions of recrossing and tunneling effects. These values are very close to the experimentally deduced one (14.2 kcal·mol−1) and ~2 kcal·mol−1 smaller than the ones obtained with the closed loop conformer. Calculation of primary KIEs and IR spectra in both protein conformations are also consistent with our hypothesis and in agreement with experimental data. Our calculations suggest that the closure of the active site is mainly required for the inverse process; the oxidation of lactate to pyruvate. According to this hypothesis H4 type LDH enzyme molecules, where it has been propose that lactate is transformed into pyruvate, should have a better ability to close the mobile loop than the M4 type LDH molecules. PMID:25705562

  14. Parathyroid hormone-related protein blood test

    MedlinePlus

    ... gov/ency/article/003691.htm Parathyroid hormone-related protein blood test To use the sharing features on ... page, please enable JavaScript. The parathyroid hormone-related protein (PTH-RP) test measures the level of a ...

  15. Effects of dietary forage sources on rumen microbial protein synthesis and milk performance in early lactating dairy cows.

    PubMed

    Zhu, W; Fu, Y; Wang, B; Wang, C; Ye, J A; Wu, Y M; Liu, J-X

    2013-03-01

    The objective of this study was to evaluate the effects of dietary forage sources on milk performance, rumen microbial protein synthesis, and N utilization in early lactation dairy cows. Twelve primiparous Chinese Holstein dairy cows (45 ± 6.0 DIM) were used in a 3 × 3 Latin square design. Diets were isonitrogenous and isocaloric, with a forage-to-concentrate ratio of 45:55 [dry matter (DM) basis] and contained similar concentrate mixtures. Different forage sources were then added (on a DM basis): 21% corn silage, 19% corn stover, and 5% alfalfa hay (CS); 19% corn silage, 21% Chinese wild rye hay and 5% alfalfa hay (CWR); or 19% corn silage, 9% Chinese wild rye hay, and 17% alfalfa hay (AH). Each period lasted for 21 d, with the first 14 d for an adaptation period. Dry matter intake was not affected by the source of dietary forage. Milk yield was higher for cows fed AH than those fed CS, with an intermediate value for CWR. Milk protein content was higher in the cows fed AH compared with CWR (3.02 vs. 2.92%), with CS (2.95%) at an intermediate position. The contents of milk fat and lactose were not different among the treatments. However, milk efficiency (milk yield/DM intake) was higher for cows fed AH than those fed CS, with those fed CWR intermediate. Cows fed AH had higher microbial protein yield and metabolizable protein than those fed CS or CWR. The concentrations of urea N in the urine, blood, and milk were decreased for cows fed AH, indicating an increased N conversion. The results indicated that corn stover could replace Chinese wild rye grass in the diets for lactating cows and that a high proportion of alfalfa hay in the diet is beneficial for milk protein production by increasing microbial protein yield. This can be attributed to the improving the supply of rumen-available energy.

  16. Maternal protein restriction in pregnancy and/or lactation affects seminiferous tubule organization in male rat offspring.

    PubMed

    Rodríguez-González, G L; Vigueras-Villaseñor, R M; Millán, S; Moran, N; Trejo, R; Nathanielsz, P W; Larrea, F; Zambrano, E

    2012-10-01

    Maternal protein restriction (MPR) during pregnancy impaired the reproduction of male offspring. We investigated, during the first wave of spermatogenesis, whether MPR exerts deleterious effects on germ cell proliferation and differentiation, as well as androgen receptor (AR) protein expression, which was used as a marker for Sertoli cell (SC) maturation. At the beginning of pregnancy (day 0), dams were fed a control diet (C: 20% casein) or a restricted isocaloric diet (R: 10% casein). After birth, four groups were established: CC, RR, CR and RC (first letter diet during pregnancy and second during lactation). Male offspring were studied at postnatal days 14, 21 and 36. At birth, pup body weight was unchanged. Body weight and testis weight were reduced in RR and CR groups at all ages evaluated. MPR delayed the germinal epithelium development at all ages evaluated. On performing Western blot and immunohistochemistry, AR expression was found to be lower in the three restricted groups. The results suggest that MPR during pregnancy and/or lactation delays SC maturation and germ cell differentiation, and affects intratubular organization. These changes might be responsible for the lower fertility rate at older ages.

  17. Effect of dietary crude protein level and degradability on ruminal fermentation and nitrogen utilization in lactating dairy cows.

    PubMed

    Hristov, A N; Etter, R P; Ropp, J K; Grandeen, K L

    2004-11-01

    The objectives of this experiment were to investigate the effects of two ruminally degradable protein (RDP) levels in diets containing similar ruminally undegradable protein (RUP) and metabolizable protein (MP) concentrations on ruminal fermentation, digestibility, and transfer of ruminal ammonia N into milk protein in dairy cows. Four ruminally and duodenally cannulated Holstein cows were allocated to two dietary treatments in a crossover design. The diets (adequate RDP [ARDP] and high RDP [HRDP]), had similar concentrations of RUP and MP, but differed in CP/RDP content. Ruminal ammonia was labeled with 15N and secretion of tracer in milk protein was determined for a period of 120 h. Ammonia concentration in the rumen tended to be greater (P = 0.06) with HRDP than with ARDP. Microbial N flow to the duodenum, ruminal digestibility of dietary nutrients, DMI, milk yield, fat content, and protein content and yield were not statistically different between diets. There was a tendency (P = 0.07) for increased urinary N excretion, and blood plasma and milk urea N concentrations were greater (P = 0.002 and P = 0.01, respectively) with HRDP compared with ARDP. Milk N efficiency was decreased (P = 0.01) by the HRDP diet. The cumulative secretion of ammonia 15N into milk protein, as a proportion of 15N dosed intraruminally, was greater (P = 0.003) with ARDP than with HRDP. The proportions of bacterial protein originating from ammonia N and milk protein originating from bacterial or ammonia N averaged 43, 61, and 26% and were not affected by diet. This experiment indicated that excess RDP in the diet of lactating dairy cows could not be efficiently utilized for microbial protein synthesis and was largely lost through urinary N excretion. At a similar MP supply, increased CP or RDP concentration of the diet would result in decreased efficiency of conversion of dietary N into milk protein and less efficient use of ruminal ammonia N for milk protein syntheses.

  18. Parathyroid hormone-related protein is a gravisensor in lung and bone cell biology

    NASA Astrophysics Data System (ADS)

    Torday, J. S.

    2003-10-01

    Parathyroid Hormone-related Protein (PTHrP) has been shown to be essential for the development and homeostatic regulation of lung and bone. Since both lung and bone structure and function are affected by microgravity, we hypothesized that 0 × g down-regulates PTHrP signaling. To test this hypothesis, we suspended lung and bone cells in the simulated microgravity environment of a Rotating Wall Vessel Bioreactor, which simulates microgravity, for up to 72 hours. During the first 8 hours of exposure to simulated 0 × g, PTHrP expression fell precipitously, decreasing by 80-90%; during the subsequent 64 hours, PTHrP expression remained at this newly established level of expression. PTHrP production decreased from 12 pg/ml/hour to 1 pg/ml/hour in culture medium from microgravity-exposed cells. The cells were then recultured at unit gravity for 24hours, and PTHrP expression and production returned to normal levels. Based on these findings, we have obtained bones from rats flown in space for 2 weeks (Mission STS-58, SL-2). Analysis of PTHrP expression by femurs and tibias from these animals (n=5) revealed that PTHrP expression was 60% lower than in bones from control ground-based rats. Interestingly, there were no differences in PTHrP expression by parietal bone from space-exposed versus ground-based animals, indicating that the effect of weightlessness on PTHrP expression is due to the unweighting of weight-bearing bones. This finding is consistent with other studies of microgravity-induced osteoporosis. The loss of the PTHrP signaling mechanism may be corrected using chemical agents that up-regulate this pathway. In conclusion, PTHrP represents a stretch-sensitive paracrine signaling mechanism that may sense gravity.

  19. Use of two different dietary energy and protein contents to define nutritive requirements of lactating buffalo cows.

    PubMed

    Bovera, F; Cutrignelli, M I; Calabrò, S; Piccolo, G; Tudisco, R; D'Urso, S; Infascelli, F

    2007-06-01

    Two total mixed rations, differing in energy and protein concentrations (diet A: 6.1 MJ/kg DM of NE(l), 112.5 g/kg DM of metabolizable protein, MP, and 101.7 g of protein digestible at the intestinal level, PDI, vs. diet B: 6.4 MJ, 95.1 g of MP, 78.9 g of PDI) were administered ad libitum for 1 year to two groups of 12 buffaloes in which the partial monthly turnover of animals allowed the lactation stage to be maintained close to 160 days. The diets were formulated to obtain, with a daily intake of about 15.5 kg of DM, a balanced energy supply but an excess of protein (diet A); the opposite occurred with diet B. The average dry matter intake recorded during the trial was 14.5 kg for diet A and 15.0 kg for diet B. Milk production was similar for both groups (7.94 kg vs. 7.95 kg). The diet with a higher protein concentration brought about a significant increase in protein, casein and non-protein nitrogen contents in the milk. A significant increase in milk fat percentage was observed when the animals were fed the diet with a higher energy concentration. The serum urea concentration (8.62 mm/l) of buffaloes fed diet A was outside the physiological range for lactating buffaloes. The energy and protein conversion efficiencies for milk production were more favourable when their supplies were balanced: 3.58 MJ of NE(l)/kg of FPCM for diet A; 2.50 g of MP or 2.07 g of PDI/g of protein milk for diet B. Moreover, when the requirements were satisfied, greater energy or protein supplies failed to induce significant changes in milk yield and quality. Finally, regarding the great differences between estimated MP requirements and those recorded during the trial, the authors suggest that the equations used in the CPM-Dairy software, formulated for dairy cows, are not applicable in buffaloes.

  20. Negative calcium balance during lactation in rural Mexican women.

    PubMed

    DeSantiago, Soledad; Alonso, Leticia; Halhali, Ali; Larrea, Fernando; Isoard, Fernando; Bourges, Héctor

    2002-10-01

    Additional calcium is required during lactation, and several calcium regulatory factors are involved in calcium balance. In lactating rural women who have marginal nutrition and consume a high-fiber diet, negative calcium balance may be expected. We evaluated calcium balance and its association with potential calcium regulatory factors in lactating, rural Mexican women who had marginal nutrition and consumed a high-fiber diet. This cross-sectional study included women at 1, 3, 6, and 12 mo of lactation (L1, L3, L6, and L12 groups) and women who had weaned their infants (W group). Age-matched, nonlactating women (NL group) were also included. Calcium balance and concentrations of calcium regulatory factors were determined. Correlation analysis was performed by using data from all of the lactating women. Calcium balance in the L1, L3, and L6 groups was negative and was significantly different (P < 0.05) from that in the W and NL groups. Serum parathyroid hormone (PTH) and 1,25-dihydroxyvitamin D [1,25-(OH)(2)D] concentrations were significantly higher (P < 0.05) in the W group than in the L and NL groups. Calcium balance was positively associated with serum estradiol concentrations (r = 0.58, P < 0.05) and negatively associated with serum 1,25-(OH)(2)D concentrations (r = -0.52, P < 0.05). Breast-milk calcium concentrations correlated positively with serum PTH-related peptide (PTHrP) concentrations (r = 0.51, P < 0.05) and negatively with serum estradiol concentrations (r = -0.57, P < 0.05). Negative calcium balance was observed during lactation in rural Mexican women who consumed a high-fiber diet. Furthermore, the data suggest that the hormones estradiol and PTHrP are involved in the regulation of calcium balance and of the calcium content of milk during lactation.

  1. Expression of parathyroid hormone-related protein confers malignant potential to mucoepidermoid carcinoma

    PubMed Central

    NAGAMINE, KYOSUKE; KITAMURA, TETSUYA; YANAGAWA-MATSUDA, AYA; OHIRO, YOICHI; TEI, KANCHU; HIDA, KYOKO; HIGASHINO, FUMIHIRO; TOTSUKA, YASUNORI; SHINDOH, MASANOBU

    2013-01-01

    Parathyroid hormone-related protein (PTHrP) is known to induce bone resorption by activating RANKL as well as PTH. PTHrP plays a central role in humoral hypercalcemia, and its expression has been reported to be closely associated with bone metastasis of breast carcinoma. PTHrP expression in oral squamous carcinoma cell lines was investigated, and PTHrP was expressed in oral squamous cell carcinoma cell lines similar to that in a prostate carcinoma cell line. Mucoepidermoid carcinoma is the most common malignant salivary gland tumor composed of different types of cells including a squamous component. Its clinical behavior is highly variable and ranges from slow-growing and indolent to locally aggressive and highly metastatic. We examined the PTHrP expression in mucoepidermoid carcinoma and assessed the significance of its correlation with clinicopathological features. Immunohistochemical detection of PTHrP was carried out in 21 cases of mucoepidermoid carcinoma in the head and neck region. PTHrP was highly detectable in intermediate and epidermoid cells, and abundant expression of PTHrP in intermediate cells had a significant association with cancer malignancy, including lymph node metastasis and/or tumor recurrence. These results suggest that PTHrP expression can be used as a prognostic factor for mucoepidermoid carcinoma. PMID:23588777

  2. Effect of protein source and soluble carbohydrate addition on rumen fermentation and lactation performance of Holstein cows.

    PubMed

    McCormick, M E; Redfearn, D D; Ward, J D; Blouin, D C

    2001-07-01

    Rumen in vitro and in vivo experiments were conducted to evaluate the effect of rumen undegradable protein and soluble carbohydrates on rumen ammonia N release and lactation performance of Holstein cows. In the in vitro experiment, freeze-dried annual ryegrass (Lolium multiflorum, LAM) pasture was supplemented 1:1 with ground corn-based grain supplements containing expeller or solvent soybean meal with sucrose or lactose supplements at 0, 2.5, or 5% of dry matter (DM). The ammonia release rate was slower with expeller compared with solvent soybean meal-supplemented diets. Sucrose supplementation at the 5% level lowered rumen ammonia concentrations, but lactose-fortification of grain supplements was without effect. In the in vivo study, 32 multiparous Holstein cows were blocked according to milk yield and randomly assigned to corn-based grain supplements containing 1) solvent soybean meal, 2) solvent soybean meal + 5% sucrose supplement, 3) expeller soybean meal, or 4) expeller soybean meal + 5% sucrose supplement. Grain supplements and fresh annual ryegrass were component fed at approximately a 1:1 grain to forage ratio (DM basis). Forage DM intake was higher for cows receiving solvent soybean meal supplemented grain supplements than those receiving expeller soybean meal (12.2 +/- 2.1 vs. 11.4 +/- 2.2 kg/d), but total DM intake was similar for all diets (22.8 +/- 2.9 kg/d). Fat-corrected milk yield was similar for all diets averaging 37.5, 38.2, 39.1, and 37.6 kg/d for diets 1 to 4, respectively. Rumen fermentation, milk urea nitrogen, and body condition were unaffected by supplements; however, cows fed grain supplement 1 utilized dietary energy more efficiently than cows offered the other dietary treatments. High dietary crude protein concentrations may have limited lactation response to rumen undegradable protein and sugar.

  3. A method to estimate cow potential and subsequent responses to energy and protein supply according to stage of lactation.

    PubMed

    Daniel, J B; Friggens, N C; Van Laar, H; Ferris, C P; Sauvant, D

    2017-05-01

    Milk responses to dietary change are influenced by the relative production level, that is, the distance between observed production and potential production. The closer the animal is to its potential, the smaller the expected response is to extra nutrients. Therefore, the aim of this work was to provide a method to quantify cow potential, to estimate subsequent responses to changes in nutrient supply. The observed efficiencies in net energy for lactation (NEL) and metabolizable protein (MP) are proposed as a basis to estimate the relative production level of the animal. The rationale for using NEL and MP efficiency (ratios of milk energy yield/NEL above maintenance supply and milk protein yield/MP above maintenance supply) builds on the uniformity of the observed relationships between size of the milk responses and extra NEL supply and MP supply, when centered on a given efficiency. From there, a pivot nutritional situation where MP and NEL efficiency are 0.67 and 1.00, respectively, was defined, from which milk responses could be derived across animals varying in production potential. An implicit assumption of using response equations centered on reference efficiency pivots is that the size of the response to a fixed change in nutrient supply, relative to the pivot, is identical for animals with different production capacities. The proposed approach was evaluated with 2 independent data sets, where different dietary treatments were applied during the whole lactation. In these data sets, MP and NEL above maintenance supply were calculated weekly using the recently updated INRA Systali feed units system. Differences in NEL and MP supply above maintenance between the extreme dietary treatments were large, on average 667 g of MP/d and 13 MJ of NEL/d (3.11 Mcal/d) in the first data set, and 513 g of MP/d and 29 MJ of NEL/d (6.93 Mcal/d) for the second data set. Milk energy yield and milk component yields were predicted with root mean square prediction errors between 7

  4. Genetic parameters across lactation for feed intake, fat- and protein-corrected milk, and liveweight in first-parity Holstein cattle.

    PubMed

    Manzanilla Pech, C I V; Veerkamp, R F; Calus, M P L; Zom, R; van Knegsel, A; Pryce, J E; De Haas, Y

    2014-09-01

    Breeding values for dry matter intake (DMI) are important to optimize dairy cattle breeding goals for feed efficiency. However, generally, only small data sets are available for feed intake, due to the cost and difficulty of measuring DMI, which makes understanding the genetic associations between traits across lactation difficult, let alone the possibility for selection of breeding animals. However, estimating national breeding values through cheaper and more easily measured correlated traits, such as milk yield and liveweight (LW), could be a first step to predict DMI. Combining DMI data across historical nutritional experiments might help to expand the data sets. Therefore, the objective was to estimate genetic parameters for DMI, fat- and protein-corrected milk (FPCM) yield, and LW across the entire first lactation using a relatively large data set combining experimental data across the Netherlands. A total of 30,483 weekly records for DMI, 49,977 for FPCM yield, and 31,956 for LW were available from 2,283 Dutch Holstein-Friesian first-parity cows between 1990 and 2011. Heritabilities, covariance components, and genetic correlations were estimated using a multivariate random regression model. The model included an effect for year-season of calving, and polynomials for age of cow at calving and days in milk (DIM). The random effects were experimental treatment, year-month of measurement, and the additive genetic, permanent environmental, and residual term. Additive genetic and permanent environmental effects were modeled using a third-order orthogonal polynomial. Estimated heritabilities ranged from 0.21 to 0.40 for DMI, from 0.20 to 0.43 for FPCM yield, and from 0.25 to 0.48 for LW across DIM. Genetic correlations between DMI at different DIM were relatively low during early and late lactation, compared with mid lactation. The genetic correlations between DMI and FPCM yield varied across DIM. This correlation was negative (up to -0.5) between FPCM yield in

  5. Effects of feeding diets naturally contaminated with Fusarium mycotoxins on protein metabolism in late gestation and lactation of first-parity sows.

    PubMed

    Díaz-Llano, G; Smith, T K; Boermans, H J; Caballero-Cortes, C; Friendship, R

    2010-03-01

    A study was conducted to assess the effects of feeding a blend of grains naturally contaminated with Fusarium mycotoxins to sows on the capacity for protein synthesis in skeletal muscle, the protein content per cellular unit, and the efficacy of a polymeric glucomannan adsorbent (GMA) to prevent these effects in late gestation and in lactation. Thirty-two Yorkshire sows were assigned to 4 treatment groups (8 per treatment) from 91 +/- 3 d of gestation up to weaning on d 21 after farrowing. Diets included 1) control, 2) contaminated grains, and 3) contaminated grains + 0.2% GMA. A fourth treatment of feeding sows the control diet at a restricted feed allowance was also included. The variables measured include ADFI, average daily BW change, serum total protein, urea, and ammonia, and skeletal muscle DNA, RNA, and protein. To assess the capacity for protein synthesis, ratios of RNA:DNA, and RNA:protein were compared among dietary treatments. To assess the degree of muscle protein mobilization in gestation and lactation, ratios of protein:DNA were compared among dietary treatments. Muscle samples were obtained from the triceps brachii. Blood and muscle samples were obtained 3 times: the first was obtained 1 d before the sows began to receive the experimental diets (90 +/- 3 d of gestation), a second sample was obtained 14 d later (104 +/- 3 d of gestation), and the third sample was obtained 10 d after farrowing. Serum ammonia concentrations were similar in sows fed the contaminated feed and sows fed the restricted feed compared with controls, but serum ammonia concentrations were greater in sows fed contaminated feed (P = 0.02) and restricted-fed sows (P = 0.008) compared with sows fed the contaminated grains plus GMA on 104 +/- 3 d of gestation. There were no reductions in the capacity for protein synthesis caused by mycotoxins or restricted feeding compared with controls. A reduction in ADFI (P = 0.003) was observed in sows fed the 2 contaminated diets in lactation

  6. Mammary-Specific Ablation of the Calcium-Sensing Receptor During Lactation Alters Maternal Calcium Metabolism, Milk Calcium Transport, and Neonatal Calcium Accrual

    PubMed Central

    Mamillapalli, Ramanaiah; VanHouten, Joshua; Dann, Pamela; Bikle, Daniel; Chang, Wenhan; Brown, Edward

    2013-01-01

    To meet the demands for milk calcium, the lactating mother adjusts systemic calcium and bone metabolism by increasing dietary calcium intake, increasing bone resorption, and reducing renal calcium excretion. As part of this adaptation, the lactating mammary gland secretes PTHrP into the maternal circulation to increase bone turnover and mobilize skeletal calcium stores. Previous data have suggested that, during lactation, the breast relies on the calcium-sensing receptor (CaSR) to coordinate PTHrP secretion and milk calcium transport with calcium availability. To test this idea genetically, we bred BLG-Cre mice with CaSR-floxed mice to ablate the CaSR specifically from mammary epithelial cells only at the onset of lactation (CaSR-cKO mice). Loss of the CaSR in the lactating mammary gland did not disrupt alveolar differentiation or milk production. However, it did increase the secretion of PTHrP into milk and decreased the transport of calcium from the circulation into milk. CaSR-cKO mice did not show accelerated bone resorption, but they did have a decrease in bone formation. Loss of the mammary gland CaSR resulted in hypercalcemia, decreased PTH secretion, and increased renal calcium excretion in lactating mothers. Finally, loss of the mammary gland CaSR resulted in decreased calcium accrual by suckling neonates, likely due to the combination of increased milk PTHrP and decreased milk calcium. These results demonstrate that the mammary gland CaSR coordinates maternal bone and calcium metabolism, calcium transport into milk, and neonatal calcium accrual during lactation. PMID:23782944

  7. Analysis of production responses to changing crude protein levels in lactating dairy cow diets when evaluated in continuous or change-over experimental designs

    USDA-ARS?s Scientific Manuscript database

    The objective of this study was to evaluate by meta-analysis the effect of experimental design on the production response functions obtained when changing crude protein levels in lactating dairy cow diets. The final database of studies that met the selection criteria contained 55 publications with 2...

  8. Cyclic AMP and AKAP-mediated targeting of protein kinase A regulates lactate dehydrogenase subunit A mRNA stability.

    PubMed

    Jungmann, Richard A; Kiryukhina, Olga

    2005-07-01

    Expression of the lactate dehydrogenase A subunit (ldh-A) gene is controlled through transcriptional as well as post-transcriptional mechanisms. Both mechanisms involve activation of protein kinase A (PKA) into its subunits and subsequent phosphorylation and activation of several key regulatory factors. In rat C6 glioma cells, post-transcriptional gene regulation occurs through PKA-mediated stabilization of LDH-A mRNA and subsequent increase of intracellular LDH-A mRNA levels. Previous studies have demonstrated a cAMP-stabilizing region (CSR) located in the LDH-A 3'-untranslated region which, in combination with several phosphorylated CSR-binding proteins (CSR-BP), regulates the PKA-mediated stabilization of LDH-A mRNA. However, the mechanistic details of interaction of CSR with proteins as they pertain to mRNA stabilization by PKA are so far largely unknown. In this study we tested the hypothesis that ribosomal protein extracts (RSW) from glioma cells contain PKA regulatory (RII) and catalytic (C) subunits that, in combination with a protein kinase A anchoring protein (AKAP 95) and CSR-BPs participate in forming CSR-protein complexes that are responsible for mRNA stability regulation. To demonstrate the importance of CSR-protein complex formation, the PKA subunits and AKAP 95 were removed from the RSW by immunoprecipitation, and the antigen-deleted RSW were subjected to CSR binding analysis using gel mobility shift and UV cross-linking. It was shown that AKAP 95 as well as RII formed a direct linkage with CSR during CSR-protein complex formation. In contrast, the catalytic subunit formed part of the CSR-protein complex but did not bind to CSR directly in a covalent linkage. To determine whether formation of CSR complexes that included C, RII, and AKAP 95 constituted a functional event and was necessary for mRNA stabilization, cell-free decay reactions were carried out with RSW extracts, and the kinetics of decay of LDH-A mRNA was determined. Depletion of PKA

  9. PTHrP Overexpression Partially Inhibits a Mechanical Strain-Induced Arthritic Phenotype in Chondrocytes

    PubMed Central

    Wang, Dean; Taboas, Juan M.; Tuan, Rocky S.

    2010-01-01

    Objective Cell-based tissue engineering strategies are currently in clinical use and continue to be developed at a rapid pace for the repair of cartilage defects. Regardless of the repair methodology, chondrocytes within newly regenerated cartilage remain susceptible to the abnormal inflammatory and mechanical environments that underlie osteoarthritic disease, likely compromising the implant’s integration, function, and longevity. The present study investigates the use of parathyroid hormone-related peptide (PTHrP) overexpression for chondroprotection. Design Bovine articular chondrocytes were transfected with human PTHrP (hPTHrP) constructs (1-141 or 1-173) and subjected to injurious cyclic tensile strain (CTS; 0.5 Hz and 16% elongation) for 48 hours. mRNA expression of matrix remodeling, inflammatory signaling, hypertrophic, and apoptotic genes were examined with real-time reverse transcription polymerase chain reaction. Nitric oxide (NO) and prostaglandin E2 (PGE2) production were measured using the Griess assay and enzyme immunoassay, respectively. Results CTS induced an arthritic phenotype in articular chondrocytes as indicated by increased gene expression of collagenases and aggrecanases and increased production of NO and PGE2. Additionally, CTS increased collagen type X (Col10a1) mRNA expression, whereas overexpression of either hPTHrP isoform inhibited CTS-induced Col10a1 gene expression. However, hPTHrP 1-141 augmented CTS-induced NO and PGE2 production, and neither hPTHrP isoform had any significant effect on apoptotic genes. Conclusions Our results suggest that chondrocytes overexpressing PTHrP resist mechanical strain-induced hypertrophic-like changes. Therapeutic PTHrP gene transfer may be considered for chondroprotection applications in newly regenerated cartilage. PMID:21087676

  10. Reaction norm of fertility traits adjusted for protein and fat production level across lactations in Holstein cattle.

    PubMed

    Menendez-Buxadera, A; Carabaño, M J; Gonzalez-Recio, O; Cue, R I; Ugarte, E; Alenda, R

    2013-07-01

    A total of 304,001 artificial insemination outcomes in up to 7 lactations from 142,389 Holstein cows, daughters of 5,349 sires and 101,433 dams, calving between January 1995 and December 2007 in 1,347 herds were studied by a reaction norm model. The (co)variance components for days to first service (DFS), days open, nonreturn rate in the first service (NRFS), and number of services per conception were estimated by 6 models: 3 Legendre polynomial degrees for the genetic effects and adjustment or not for the level of fat plus protein (FP) production recorded at day closest to DFS. For all traits and type of FP adjustment, a second degree polynomial showed the best fit. The use of the adjusted FP model did not increase the level of genetic (co)variance components except for DFS. The heritability for each of the traits was low in general (0.03-0.10) and increased from the first to fourth calving; nevertheless, very important variability was found for the estimated breeding value (EBV) of the sires. The genetic correlations (rg) were close to unity between adjacent calvings, but decreased for most distant parities, ranging from rg=0.36 (for DFS) to rg=0.63 (for NRFS), confirming the existence of heterogeneous genetic (co)variance components and EBV across lactations. The results of the eigen decomposition of rg shows that the first eigenvalue explained between 82 to 92% and the second between 8 to 14% of the genetic variance for all traits; therefore, a deformation of the overall mean trajectory for reproductive performance across the trajectory of the different calving could be expected if selection favored these eigenfunctions. The results of EBV for the 50 best sires showed a substantial reranking and variation in the shape of response across lactations. The more important aspect to highlight, however, is the difference between the EBV of the same sires in different calvings, a characteristic known as plasticity, which is particularly important for DFS and NRFS. This

  11. Influence of protein percentage and degradability on performance of lactating cows during moderate temperature.

    PubMed

    Higginbotham, G E; Huber, J T; Wallentine, M V; Johnston, N P; Andrus, D

    1989-07-01

    Sixty high producing Holstein cows (15/treatment) averaging 157 d postpartum were offered the following diets: high protein (18.5%), high rumen degradability (60%); high protein (18.0%), medium degradability (41%); medium protein (15.4%), high degradability (61%); medium protein (15.0%), medium degradability (46%). All diets contained 32% corn silage, 15% alfalfa haylage, and 10% whole cottonseed. Degradabilities were determined by the ficin method and were lowered by replacing soybean meal with meat and bone meal and brewers dried grains. Cows were milked three times daily and the study was conducted from May 11 to June 23 in Provo, UT during ascending day temperatures but cool nights. Milk yields were not different for the respective treatments (36.9, 35.4, 34.8, and 36.7 kg/d), but milk fat was higher at high protein degradability: 3.11, 2.89, 3.04, 2.78%, respectively, resulting in a tendency toward higher FCM. Lower acetate and propionate and acetate:propionate ratios were observed for medium degradability diets and were consistent with lower milk fat. Rumen NH3, butyrate, valerate, isovalerate, 2-methyl butyrate, and blood urea N were higher for cows on high protein diets but were not affected by degradability. Serum glucose and cortisol were not significantly affected by protein treatment, but serum triiodothyronine was lower on high protein; the interaction effect for thyroxine was significant.

  12. Dissecting the impact of protein versus energy host nutrition on the expression of immunity to gastrointestinal parasites during lactation.

    PubMed

    Jones, L A; Houdijk, J G M; Sakkas, P; Bruce, A D; Mitchell, M; Knox, D P; Kyriazakis, I

    2011-06-01

    Many mammals exhibit a periparturient relaxation of previously established immune responses (PPRI) to gastrointestinal nematodes culminating in increased worm burdens. It has been suggested that the extent of PPRI may have a nutritional basis as it is considerably augmented when protein supply is scarce. Subsequent studies have shown that increased dietary protein intake can ameliorate this phenomenon. However, this effect is often confounded with increased food intake and thus increased energy levels. Herein, we aimed to dissect the effects of protein and energy nutrition on the immune status and resistance to re-infection with gastrointestinal nematodes in the periparturient host. The lactating, Nippostrongylus brasiliensis re-infected rat was utilised as an established model for mammalian PPRI. Experimental animals were assigned to restricted feeding regimens designed to achieve four pre-determined levels of crude protein (CP) at one of two levels of metabolisable energy (ME) and parasitological and immunological measurements taken at either day 6 or day 9 post re-infection. We clearly show that increased supply of dietary CP, but not increased dietary ME, significantly reduced worm burdens. The increased magnitude of worm expulsion with increased dietary CP supply strongly correlated with mucosal mast cell accumulation in the small intestine. In addition, increased CP and not ME supply increased mucosal eosinophil numbers. Furthermore, increased CP led to higher levels of total IgG at high ME only and there were interactive effects of CP and ME on serum levels of IgG1 and IgG2a. Perhaps surprisingly, CP nutrition did not affect expression of either Th1 (IFN-γ) or Th2 (IL-4, IL-13) cytokines in the mesenteric lymph nodes. These data emphasise the role of immunonutrition, and particularly dietary protein, in combating infectious disease such as gastrointestinal parasitism. Copyright © 2011 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All

  13. Effect of dietary level of rumen-degraded protein on production and nitrogen metabolism in lactating dairy cows.

    PubMed

    Reynal, S M; Broderick, G A

    2005-11-01

    Twenty-eight (8 with ruminal cannulas) lactating Holstein cows were assigned to 4 x 4 Latin squares and fed diets with different levels of rumen-degraded protein (RDP) to study the effect of RDP on production and N metabolism. Diets contained [dry matter (DM) basis] 37% corn silage, 13% alfalfa silage, and 50% concentrate. The concentrate contained solvent and lignosulfonate-treated soybean meal and urea, and was adjusted to provide RDP at: 13.2, 12.3, 11.7, and 10.6% of DM in diets A to D, respectively. Intake of DM and yield of milk, fat-corrected milk, and fat were not affected by treatments. Dietary RDP had positive linear effects on milk true protein content and microbial non-ammonia N (NAN) flow at the omasal canal, and a quadratic effect on true protein yield, with maximal protein production at 12.3% RDP. However, dietary RDP had a positive linear effect on total N excretion, with urinary N accounting for most of the increase, and a negative linear effect on environmental N efficiency (kg of milk produced per kg of N excreted). Therefore, a compromise between profitability and environmental quality was achieved at a dietary RDP level of 11.7% of DM. Observed microbial NAN flow and RDP supply were higher and RUP flow was lower than those predicted by the NRC (2001) model. The NRC (2001) model overpredicted production responses to RUP compared with the results in this study. Replacing default NRC degradation rates for protein supplements with rates measured in vivo resulted in similar observed and predicted values, suggesting that in situ degradation rates used by the NRC are slower than apparent rates in this study.

  14. Identification of parathyroid hormone-related protein-derived peptides immunogenic in human histocompatibility leukocyte antigen-A24+ prostate cancer patients.

    PubMed

    Yao, A; Harada, M; Matsueda, S; Ishihara, Y; Shomura, H; Noguchi, M; Matsuoka, K; Hara, I; Kamidono, S; Itoh, K

    2004-07-19

    Parathyroid hormone-related protein (PTHrP) is a key factor in the development of bone metastases, which are a major barrier in treating prostate cancer patients. In this study, we attempted to identify PTHrP-derived peptides immunogenic in human histocompatibility leukocyte antigen (HLA)-A24(+) prostate cancer patients. Among four different PTHrP peptides carrying the HLA-A24 binding motif, both the PTHrP(36-44) and PTHrP(102-111) peptides efficiently induced peptide-specific cytotoxic T lymphocytes from peripheral blood mononuclear cells (PBMCs) of HLA-A24(+) prostate cancer patients. Peptide-stimulated PBMCs showed cytotoxicity against prostate cancer cells in an HLA-A24-restricted manner. Experiments using antibodies and cold inhibition targets confirmed that their cytotoxicity was dependent on PTHrP peptide-specific and CD8(+) T cells. Immunoglobulin G reactive to the PTHrP(102-111) or PTHrP(110-119) peptide was frequently detected in the plasma of prostate cancer patients, suggesting that the PTHrP(102-111) peptide is able to elicit cellular and humoral immune responses in cancer patients. These results indicate that the PTHrP could be a promising target molecule for specific immunotherapy of HLA-A24(+) prostate cancer patients with metastases.

  15. Effects of decreasing metabolizable protein and rumen-undegradable protein on milk production and composition and blood metabolites of Holstein dairy cows in early lactation.

    PubMed

    Bahrami-Yekdangi, H; Khorvash, M; Ghorbani, G R; Alikhani, M; Jahanian, R; Kamalian, E

    2014-01-01

    This study was conducted to evaluate the effects of decreasing dietary protein and rumen-undegradable protein (RUP) on production performance, nitrogen retention, and nutrient digestibility in high-producing Holstein cows in early lactation. Twelve multiparous Holstein lactating cows (2 lactations; 50 ± 7 d in milk; 47 kg/d of milk production) were used in a Latin square design with 4 treatments and 3 replicates (cows). Treatments 1 to 4 consisted of diets containing 18, 17.2, 16.4, and 15.6% crude protein (CP), respectively, with the 18% CP diet considered the control group. Rumen-degradable protein levels were constant across the treatments (approximately 10.9% on a dry matter basis), whereas RUP was gradually decreased. All diets were calculated to supply a postruminal Lys:Met ratio of about 3:1. Dietary CP had no significant effects on milk production or milk composition. In fact, 16.4% dietary CP compared with 18% dietary CP led to higher milk production; however, this effect was not significant. Feed intake was higher for 16.4% CP than for 18% CP (25.7 vs. 24.3 kg/d). Control cows had greater CP and RUP intakes, which resulted in higher concentrations of plasma urea nitrogen and milk urea nitrogen; cows receiving 16.4 and 15.6% CP, respectively, exhibited lower concentrations of milk urea nitrogen (15.2 and 15.1 vs. 17.3 mg/dL). The control diet had a significant effect on predicted urinary N. Higher CP digestibility was recorded for 18% CP compared with the other diets. Decreasing CP and RUP to 15.6 and 4.6% of dietary dry matter, respectively, had no negative effects on milk production or composition when the amounts of Lys and Met and the Lys:Met ratio were balanced. Furthermore, decreasing CP and RUP to 16.4 and 5.4%, respectively, increased dry matter intake. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  16. Efficacy of using a combination of rendered protein products as an undegradable intake protein supplement for lactating, winter-calving, beef cows fed bromegrass hay.

    PubMed

    Encinias, A M; Lardy, G P; Leupp, J L; Encinias, H B; Reynolds, L P; Caton, J S

    2005-01-01

    Seventy-two (36 in each of two consecutive years) lactating, British-crossbred cows (609 +/- 19 kg) were used to evaluate effects of feeding a feather meal-blood meal combination on performance by beef cows fed grass hay. Bromegrass hay (9.6% CP, DM basis) was offered ad libitum and intake was measured daily in individual Calan electronic headgates. Acclimation to Calan gates began approximately 20 d after parturition, and treatments were initiated 21 d later. Cows were assigned randomly to one of four treatments (DM basis) for 60 d: 1) nonsupplemented control (CON), 2) energy control (ENG; 790 g/d; 100% beet pulp), 3) degradable intake protein (DIP; 870 g/d; 22% beet pulp and 78% sunflower meal), or 4) undegradable intake protein (UIP; 800 g/d; 62.5% sunflower meal, 30% hydrolyzed feather meal, and 7.5% blood meal). Net energy concentrations of supplements were formulated to provide similar NE(m) intakes (1.36 Mcal/d). The DIP and UIP supplements were calculated to supply similar amounts of DIP (168 g/d) and to supply 64 and 224 g/d of UIP, respectively. Forage DMI (kg/d) decreased in supplemented vs. nonsupplemented (P = 0.03) and DIP vs. UIP (P = 0.001); however, when expressed as a percentage of BW, forage DMI was not different (P = 0.23). Supplemented cows tended (P = 0.17) to lose less BW than CON. Body condition change was not affected (P = 0.60) by postpartum supplementation. No differences were noted in milk production (P = 0.29) or in calf gain during the supplementation period (P = 0.74). Circulating insulin concentrations were not affected by treatment (P = 0.42). In addition, supplementation did not affect circulating concentrations of NEFA (P = 0.18) or plasma urea nitrogen (P = 0.38). Results of the current study indicate that supplementation had little effect on BW, BCS, milk production, or calf BW when a moderate-quality forage (9.6% CP) was fed to postpartum, winter-calving cows in optimal body condition (BCS > 5). Supplemental UIP did not enhance

  17. Short communication: Casein hydrolysate and whey proteins as excipients for cyanocobalamin to increase intestinal absorption in the lactating dairy cow.

    PubMed

    Artegoitia, V M; de Veth, M J; Harte, F; Ouellet, D R; Girard, C L

    2015-11-01

    Bioavailability of vitamin B12 is low in humans and animals. Improving vitamin B12 absorption is important for optimal performance in dairy cows and for increasing vitamin B12 concentrations in milk for human consumption. However, when supplemented in the diet, 80% of synthetic vitamin B12, cyanocobalamin (CN-CBL), is degraded in the rumen of dairy cows and only 25% of the amount escaping destruction in the rumen disappears from the small intestine between the duodenal and ileal cannulas. In pigs, vitamin B12 from milk is more efficiently absorbed than synthetic CN-CBL. The objective of this study was to determine the efficacy of casein hydrolysate and whey proteins as excipients for CN-CBL to increase portal-drained viscera (PDV) flux of the vitamin in lactating dairy cows. Four multiparous lactating Holstein cows (237 ± 17 DIM) equipped with a rumen cannula and catheters in the portal vein and a mesenteric artery were used in a randomized Youden square design. They were fed every 2 h to maintain steady digesta flow. On experimental days, they received a postruminal bolus of (1) CN-CBL alone (0.1 g), (2) CN-CBL (0.1 g) + casein hydrolysate (10 g), or (3) CN-CBL (0.1 g) + whey proteins (10 g). Starting 30 min after the bolus, blood samples were taken simultaneously from the 2 catheters every 15 min during the first 2 h and then every 2 h until 24 h postbolus. Milk yield, DMI, and vitamin B12 portal-arterial difference and PDV flux were analyzed using the MIXED procedure of SAS. Milk yield and DMI were not affected by treatments. The portal-arterial difference of vitamin B12 during the 24-h period following the bolus of vitamin was greater when the vitamin was given in solution with casein hydrolysate (2.9 ± 4.6 pg/mL) than alone (-17.5 ± 5.2 pg/mL) or with whey protein (-13.4 ± 4.2 pg/mL). The treatment effects were similar for the PDV flux. The present results suggest that CN-CBL given with casein hydrolysate increases vitamin B12 absorption as compared with

  18. Longitudinal analysis of protein glycosylation and β-casein phosphorylation in term and preterm human milk during the first 2 months of lactation.

    PubMed

    Molinari, Claire E; Casadio, Ylenia S; Hartmann, Ben T; Arthur, Peter G; Hartmann, Peter E

    2013-07-14

    Human milk proteins provide term and preterm infants with both nutrition and protection. The objective of the present study was to examine longitudinal changes in the protein composition of term and preterm milk during the first 2 months of lactation, focusing on protein phosphorylation and glycosylation. Using gel electrophoresis, the relative concentration and glycosylation status of lactoferrin, secretory Ig A, β-casein, α-lactalbumin, serum albumin, bile salt-stimulated lipase, xanthine oxidoreductase, tenascin and macrophage mannose receptor 1 were measured in milk collected on days 7, 10, 14, 18, 21, 28 and 60 postpartum from preterm mothers (28-32 weeks gestation, n 17). The phosphorylation status of β-casein was also investigated. To determine if these variables differ in term and preterm milk, samples from term mothers (38-41 weeks gestation, n 8) collected on days 7, 14 and 30 of lactation were also analysed. The concentration of the abundant milk proteins decreased during lactation in term and preterm milk (P <0·05). No difference in protein glycosylation was observed, except for the glycoproteins serum albumin and tenascin. The phosphorylation of β-casein varied significantly between term and preterm milk. Further investigation is required to determine whether these modifications affect protein function and are clinically important to preterm infants.

  19. Effects of dietary protein solubility on nitrogen losses from lactating dairy cows and comparison with predictions from the Cornell Net Carbohydrate and Protein System.

    PubMed

    Haig, P A; Mutsvangwa, T; Spratt, R; McBride, B W

    2002-05-01

    This experiment determined the effects of dietary protein solubility on amount, form, and route of nitrogen loss in lactating Holstein dairy cows, and the ability of the Cornell Net Carbohydrate and Protein System (CNCPS) to accurately predict rumen microbial yield, serum urea N (SUN), milk urea N (MUN), and fecal N. Eighteen multiparous Holstein cows were assigned randomly to one of three dietary treatments that were similar in crude protein (17.7%) content but differed in their content of soluble intake protein (SIP). Dietary contents of SIP, as % of total CP were 30, 36, and 48%. The experimental period was 21 d, and total N balance collections were done during the last 5 d. As dietary content of SIP increased, excretion of urinary N increased quadratically, and it was the primary route of N excretion. Urinary excretion of purine derivatives (PD) responded quadratically as dietary SIP content increased. The CNCPS predicted a quadratic decrease in total metabolizable protein (MP) supply. No effect of dietary content of SIP was detected on MUN and SUN. The CNCPS predicted a quadratic decrease in SUN and MUN as dietary SIP increased. Results from this study indicated that changing the dietary content of SIP altered routes of N excretion in dairy cows, but had no effect on total N balance. The CNCPS did not adequately predict changes in SUN and MUN for cows fed diets varying in SIP.

  20. C-reactive protein is differentially modulated by co-existing infections, vitamin deficiencies and maternal factors in pregnant and lactating indigenous Panamanian women.

    PubMed

    González-Fernández, Doris; Pons, Emérita Del Carmen; Rueda, Delfina; Sinisterra, Odalis Teresa; Murillo, Enrique; Scott, Marilyn E; Koski, Kristine G

    2017-06-02

    The usefulness of C-reactive protein (CRP) as a non-specific marker of inflammation during pregnancy and lactation is unclear in impoverished populations where co-existing infections and vitamin deficiencies are common. This cross-sectional study in Panama recruited 120 pregnant and 99 lactating Ngäbe-Buglé women from 14 communities in rural Panama. Obstetric history, indoor wood smoke exposure, fieldwork, BMI, vitamins A, B12, D, and folic acid, and inflammation markers (CRP, neutrophil/lymphocyte ratio (NLR), plateletcrit and cytokines) were measured. Multiple regressions explored both associations of CRP with other inflammatory markers and associations of CRP and elevated CRP based on trimester-specific cut-offs with maternal factors, infections and vitamin deficiencies. CRP was higher in pregnancy (51.4 ± 4.7 nmol/L) than lactation (27.8 ± 3.5 nmol/L) and was elevated above trimester specific cut-offs in 21% of pregnant and 30% of lactating women. Vitamin deficiencies were common (vitamin A 29.6%; vitamin D 68.5%; vitamin B12 68%; folic acid 25.5%) and over 50% of women had two or more concurrent deficiencies as well as multiple infections. Multiple regression models highlighted differences in variables associated with CRP between pregnancy and lactation. In pregnancy, CRP was positively associated with greater indoor wood smoke exposure, caries and hookworm and negatively associated with Ascaris and vaginal Lactobacillus and Bacteroides/Gardnerella scores. Consistent with this, greater wood smoke exposure, caries as well as higher diplococcal infection score increased the odds of trimester-elevated CRP concentrations whereas longer gestational age lowered the likelihood of a trimester-elevated CRP. During lactation, folic acid deficiency was associated with higher CRP whereas parity, number of eosinophils and Mobiluncus score were associated with lower CRP. Also, a higher BMI and Trichomonas vaginalis score increased the likelihood of an elevated

  1. Lactational response of dairy cows to change of degradability of dietary protein and organic matter.

    PubMed

    Aharoni, Y; Arieli, A; Tagari, H

    1993-11-01

    The effect of variable degradability of both OM and CP, incorporated at a constant ratio in diets of high yielding dairy cows (35 kg/d), was studied under commercial dairy herd conditions. Two diets containing 17% CP were formulated, including high (70%) and low (65%) protein degradability. The ratio of rumen-degradable OM to degradable protein was adjusted to 5:1 in both diets. Cows were assigned to treatments based on equal milk yield prior to trial, parity, and DIM. The trial lasted 7 wk: a reference week (wk 0), in which both groups were fed the high degradability diet, was followed by 6 experimental wk, in which group 1 was fed the high degradability diet and group 2 the low degradability diet. Cows on the low degradability diet consumed 1.2 kg more DM and yielded 1.5 kg/d more milk, .055 kg/d more milk protein, and .196 kg/d more milk fat. Percentages of milk protein (3.06 and 3.03) were similar, but fat (3.67 and 3.28) was higher for cows fed the low degradability diet. The results suggest that, when diets were formulated to balance rumen degradability of both OM and CP, 65% rather than 70% degradability of CP was advantageous for yields of milk and milk components.

  2. Actin filament-associated protein 1 is required for cSrc activity and secretory activation in the lactating mammary gland

    PubMed Central

    Cunnick, Jess M; Kim, Stephanie; Hadsell, James; Collins, Stephen; Cerra, Carmine; Reiser, Patti; Flynn, Daniel C; Cho, Youngjin

    2014-01-01

    Actin filament-associated protein 1 (AFAP1) is an adaptor protein of cSrc that binds to filamentous actin and regulates the activity of this tyrosine kinase to affect changes to the organization of the actin cytoskeleton. In breast and prostate cancer cells, AFAP1 has been shown to regulate cellular responses requiring actin cytoskeletal changes such as adhesion, invadopodia formation and invasion. However, a normal physiological role for AFAP1 has remained elusive. In this study, we generated an AFAP1 knockout mouse model that establishes a novel physiological role for AFAP1 in lactation. Specifically, these animals displayed a defect in lactation that resulted in an inability to efficiently nurse. Histologically, the mammary glands of the lactating knockout mice were distinguished by the accumulation of large cytoplasmic lipid droplets in the alveolar epithelial cells. There was a reduction in lipid synthesis and the expression of lipogenic genes without a corresponding reduction in the production of beta-casein, a milk protein. Furthermore, these defects were associated with histological and biochemical signs of precocious involution. This study also demonstrated that AFAP1 responds to prolactin, a lactogenic hormone, by forming a complex with cSrc and becoming tyrosine phosphorylated. Together, these observations pointed to a defect in secretory activation. Certain characteristics of this phenotype mirrored the defect in secretory activation in the cSrc knockout mouse, but most importantly, the activity of cSrc in the mammary gland was reduced during early lactation in the AFAP1 null mouse and the localization of active cSrc at the apical surface of luminal epithelial cells during lactation was selectively lost in the absence of AFAP1. These data define, for the first time, the requirement of AFAP1 for the spatial and temporal regulation of cSrc activity in the normal breast, specifically for milk production. PMID:25043309

  3. Low protein diet during gestation and lactation increases food reward seeking but does not modify sucrose taste reactivity in adult female rats.

    PubMed

    da Silva, Amanda Alves Marcelino; Oliveira, Mayara Matias; Cavalcante, Taisy Cinthia Ferro; do Amaral Almeida, Larissa Cavalcanti; de Souza, Julliet Araújo; da Silva, Matilde Cesiana; de Souza, Sandra Lopes

    2016-04-01

    Nutritional deficiencies during neural development may lead to irreversible changes, even after nutritional rehabilitation, promoting morphological and functional adaptations of structures involved with various behaviours including feeding behaviour. However, the ability of the exposure low protein diet during gestation and lactation to affect the hedonic component of food intake is still poorly understood, especially in females. Wistar rats were divided into two groups according to the diet offered to the dams during pregnancy and lactation: control female (CF; diet with 17% protein, n=7) and low protein female (LPF; diet with 8% protein, n=7). The following parameters were evaluated: (a) body weight during weaning, 30, 45, 60, 75, 90 days of life; (b) standard diet intake from 110 to 132 days of life; (c) fat diet and consumption of simple carbohydrates (HFHS) for 1h at 145 days of life; (d) incentive runway task 60 days after 82 days of life; (e) taste reactivity at 90 days of life; and (f) neuronal activation in the caudate putamen, amygdala, paraventricular nucleus of the hypothalamus under stimulus HFHS at 145 days of life. The exposure, a low protein diet during gestation and lactation, decreased the body weight throughout the study period from weaning to 90 days of life. However, there was no significant change in the body weight of low protein females from 110 to 132 days of life compared with the control females. There was an increase in the rate of the search for reward and reduced the latency of the perception of bitter taste. The exposure, a low protein diet during gestation and lactation, also promoted hypophagy in adult females compared with control animals. The low protein female had increased HFHS diet consumption compared with the control. Undernutrition increased neuronal activation in response to HFHS diet consumption compared with female controls in the amygdala and in the caudate putamen. Females subjected to the exposure, a low protein diet

  4. Protein deficiency and nematode infection during pregnancy and lactation reduce maternal bone mineralization and neonatal linear growth in mice.

    PubMed

    Odiere, Maurice R; Scott, Marilyn E; Weiler, Hope A; Koski, Kristine G

    2010-09-01

    Using a 2 x 2 factorial design, we investigated the combined impact of protein deficiency (PD) and gastrointestinal nematode infection during late pregnancy and lactation on resting metabolic rate (RMR), body composition and bone mineralization, neonatal growth, and the regulatory hormones [corticosterone, leptin, and insulin-like growth factor-1 (IGF-1)] and proinflammatory cytokines [interleukin (IL)-1 beta and IL-6] that may drive these processes. Pregnant CD1 mice, fed either a protein-sufficient (PS; 24%) or protein-deficient (PD; 6%) isocaloric diet, were infected 4 times with either 0 (sham) or 100 Heligmosomoides bakeri larvae beginning on d 14 of pregnancy. Dams were killed on d 20 postpartum and pups on d 2, 7, 14, and 21. Diet and infection had largely independent effects. The PD diet elevated corticosterone and upregulated leptin concentration in maternal serum, which was associated with reduced food intake leading to lower body mass, RMR, and body temperature. Infection reduced food intake but elevated maternal serum IL-1 beta and IL-6 and did not affect corticosterone, leptin, RMR, or body temperature. The PD diet decreased maternal bone area and bone mineral content. Infection lowered maternal bone mineral density, consistent with elevated IL-1 beta and IL-6. The elevated serum IL-1 beta and lower IGF-1 in pups of PD dams and lower serum leptin and IGF-1 in pups of infected dams were both consistent with the lower pup body mass and shorter crown-rump length. This mouse model provides a novel framework to study the impact of diet and nematode infection on bone.

  5. Parathyroid hormone-related protein is a factor in normal fish pituitary.

    PubMed

    Danks, J A; Devlin, A J; Ho, P M; Diefenbach-Jagger, H; Power, D M; Canario, A; Martin, T J; Ingleton, P M

    1993-11-01

    Using antibodies to the amino-terminal region of human parathyroid hormone-related protein (PTHrP) we have demonstrated PTHrP immunoreactivity in pituitaries and plasma of the sea bream (Sparus aurata). Pituitary cells at two distinct locations contained immunodetectable PTHrP; an anterior group in the rostral pars distalis which also contained immunoreactive thyroid stimulating hormone (TSH), and a posterior group lying at the border of the pars intermedia and proximal pars distalis between cells which stained with antibody to human corticotrophin-like intermediate lobe peptide. By Western blot analysis pituitary extracts contained two immunoreactive isoforms of PTHrP, one of 29 kDa and the other of 26 kDa. Media of pituitaries incubated for up to 14 days in Krebs-Ringer bicarbonate also had several isoforms of immunodetectable PTHrP, two of them corresponding to the 29- and 26-kDa molecular forms but there were in addition both larger and smaller molecules. The concentration of PTHrP in sea bream plasma was comparable with levels observed in human subjects with humoral hypercalcaemia of malignancy. There was no reaction between pituitary cells or pituitary extracts and antibody to human parathyroid hormone. Thus sea bream pituitary contains immunoreactive PTHrP, which appears to be released into medium during in vitro incubation and which may be a significant source of plasma immunoreactive PTHrP in vivo.

  6. Gluconeogenesis, glucose handling, and structural changes in livers of the adult offspring of rats partially deprived of protein during pregnancy and lactation.

    PubMed Central

    Burns, S P; Desai, M; Cohen, R D; Hales, C N; Iles, R A; Germain, J P; Going, T C; Bailey, R A

    1997-01-01

    Maternal protein restriction is a model of fetal programming of adult glucose intolerance. Perfused livers of 48-h- starved adult offspring of rat dams fed 8% protein diets during pregnancy and lactation produced more glucose from 6 mM lactate than did control livers from rats whose dams were fed 20% protein. In control livers, a mean of 24% of the glucose formed from lactate in the periportal region of the lobule was taken up by the most distal perivenous cells; this distal perivenous uptake was greatly diminished in maternal low protein (MLP) livers, accounting for a major fraction of the increased glucose output of MLP livers. In control livers, the distal perivenous cells contained 40% of the total glucokinase of the liver; this perivenous concentration of glucokinase was greatly reduced in MLP livers. Intralobular distribution of phosphenolpyruvate carboxykinase was unaltered, though overall increased activity could have contributed to the elevated glucose output. Hepatic lobular volume in MLP livers was twice that in control livers, indicating that MLP livers had half the normal number of lobules. Fetal programming of adult glucose metabolism may operate partly through structural alterations and changes in glucokinase expression in the immediate perivenous region. PMID:9312176

  7. Energy and metabolic sensing G protein-coupled receptors during lactation-induced changes in energy balance.

    PubMed

    Friedrichs, P; Saremi, B; Winand, S; Rehage, J; Dänicke, S; Sauerwein, H; Mielenz, M

    2014-07-01

    The free fatty acid receptor FFA1, FFA2, and FFA3 and hydroxy-carboxylic acid receptor (HCA2) are G protein-coupled receptors, acting as energy and metabolic sensors. Herein, we characterized the tissue-specific mRNA abundance of genes encoding for these receptors at different stages of lactation. In addition, potential effects of supplementation with or without conjugated linoleic acids (CLA) were tested. Tissues from pluriparous cows (subcutaneous adipose tissue [SAT] and liver) and from primiparous cows (3 SAT locations, 3 visceral adipose tissues, liver, mammary gland, and skeletal muscle) were used from 2 separate trials. In primiparous cows, the mRNA abundance of all receptors (FFA3 was not detectable by the applied protocol in muscle and udder) was lowest in muscle (P < 0.05). With the exception of FFA1, gene expression of the investigated receptors was higher in adipose tissue than in the non-adipose tissue. Expression of FFA1 in liver (P < 0.03), FFAR2 in SAT (P < 0.01), and HCA2 in SAT (P < 0.01) from pluriparous cows changed during the observation period (days 21 to 252 relative to parturition). The correlation between mRNA abundance of HCA2 and peroxisome proliferator-activated receptor gamma (PPARG) and likewise PPARG2 (P < 0.01) in SAT indicates a link between HCA2 and PPARG. Differences in receptor mRNA abundance between the CLA-fed and the control animals were scarce and limited to HCA2 and FFA1 in 1 and 2 time points, respectively (less hepatic HCA2mRNA in CLA-fed pluriparous cows and greater FFA1 mRNA abundance in 2 visceral adipose tissue depots in CLA-treated primiparous cows). In view of the metabolic changes occurring during the different phases of lactation, in particular, the altered concentrations of non-esterified fatty acids and β-hydroxybutyrate acting as receptor ligands, the longitudinal tissue-specific characterization provided herein allows for a first insight into the regulation of these receptors at the gene expression level

  8. Lactation performance of dairy cows fed yeast-derived microbial protein in low- and high-forage diets.

    PubMed

    Manthey, A K; Kalscheur, K F; Garcia, A D; Mjoun, K

    2016-04-01

    The objective of this study was to investigate the effect of substituting soybean meal products with yeast-derived microbial protein (YMP) on lactation performance in diets containing 2 forage-to-concentrate ratios. Sixteen Holstein cows (4 primiparous and 12 multiparous) were randomly assigned to multiple 4 × 4 Latin squares with a 2 × 2 factorial arrangement of treatments. Diets contained low (LF; 45% of diet DM) or high forage (HF; 65% of diet DM) and YMP at 0 (NYMP) or 2.25% (WYMP) of the diet. The forage mix consisted of 67% corn silage and 33% alfalfa hay on a DM basis. No interactions of forage and YMP were noted for any of the production parameters measured. Feed efficiency (energy-corrected milk/dry matter intake) was greater for cows fed NYMP compared with WYMP. Regardless of the addition of YMP, cows fed LF had greater dry matter intake and produced more milk than cows fed HF. In addition, cows fed LF produced more energy-corrected milk than those fed HF. Milk fat percentage was lower in cows fed LF compared with HF, whereas fat yield was similar between forage concentrations. Fat yield tended to decrease with feeding YMP. Interactions of forage and YMP were observed for propionate concentration, acetate and propionate proportion, and acetate-to-propionate ratio. A tendency for an interaction of forage and YMP was also noted for ruminal pH. Cows fed HF diets had greater ruminal ammonia and butyrate concentrations, as well as proportion of butyrate. Arterial concentrations of Ile, Leu, Met, Thr, and Val were greater in cows fed LF. Cows fed NYMP had greater arterial concentrations of Ile, Lys, Trp, and Val than cows fed WYMP. Substitution of soybean proteins with YMP did not improve performance or feed efficiency of high-producing dairy cows regardless of the forage-to-concentrate ratio of the diet.

  9. Ruminal metabolism, fiber, and protein digestion by lactating cows fed calcium soap or animal-vegetable fat.

    PubMed

    Ohajuruka, O A; Wu, Z G; Palmquist, D L

    1991-08-01

    Effects of two levels of Ca soap or animal-vegetable fat on digestion of fiber and N and on microbial protein synthesis were studied in five ruminally and duodenally cannulated lactating cows in a 5 x 5 Latin square. Fat was added at 0, 2.5%, or 5% of dietary DM, which consisted of a 60:40 forage:concentrate mix. Treatment did not influence DMI, duodenal OM flow, or digestibility. Fat linearly increased liquid dilution rate (12.1 vs. 11.1 h-1) and reduced liquid (53.6 vs. 62.4 L) and ruminal (61.6 vs. 70.1 L) volumes. Source and amount of fat did not influence mean ruminal pH, ammonia N, or VFA concentrations; however, animal-vegetable fat reduced acetate:propionate ratios compared with Ca soap (3.47 vs. 3.64). Neither source nor amount of fat influenced fiber flow, digestion, or efficiency of microbial protein synthesis. Source and amount of fat did not change ruminal or total digestibilities of DM, OM, ADF, or NDF; however, dietary fat increased total tract apparent digestion of N (70.3 vs. 66.8). Estimates of ruminal DM digestibility were lower (24.2 vs. 39.0) and total tract DM digestibility was higher (63.3 vs. 60.5) with Cr marker compared with C31 hydrocarbon marker. Animal-vegetable fat or Ca soap can be supplemented up to 5% of the dietary DM in high forage diets without adverse effects on ruminal metabolism and digestion.

  10. Lactational response of dairy cows to diets varying in ruminal solubilities of carbohydrate and crude protein.

    PubMed

    Casper, D P; Schingoethe, D J

    1989-04-01

    Sixty high producing Holstein cows were randomly assigned in a 3 x 2 factorial to evaluate three sources of carbohydrates that differed in solubility and degradability (corn, barley, and dried whey) with two sources of CP solubility (soybean meal and urea) during wk 4 through 14 postpartum. Total mixed diets, formulated to be isonitrogenous at 16% CP, contained (DM basis) 40% corn silage, 10% chopped alfalfa hay, and 50% of the respective concentrate mix. Milk production (32.8, 31.5, and 31.3 kg/d) was highest for cows fed corn, whereas 4% FCM (30.0, 27.9, and 29.5 kg/d) was similar for cows fed corn and dried whey and lower for cows fed barley. Percentages of fat (3.37, 3.36, and 3.51) and protein (3.05, 3.00, and 2.98) were similar for cows fed all carbohydrate sources. Solubility of protein (soybean meal versus urea) did not affect production of milk (32.2 and 31.5 kg/d) and 4% FCM (29.4, and 28.9 kg/d). Intake of DM was lowest for cows fed barley (20.4, 18.8, and 20.5 kg/d), and intakes were similar (19.9 and 19.9 kg/d) for cows fed soybean meal and urea. Providing sources of carbohydrates in the diet that are more soluble and degradable (i.e., barley or dried whey) did not give the expected increase in utilization of a highly soluble CP source (urea) for milk production.

  11. A Comparison of Parathyroid Hormone-related Protein (1–36) and Parathyroid Hormone (1–34) on Markers of Bone Turnover and Bone Density in Postmenopausal Women: The PrOP Study

    PubMed Central

    Horwitz, Mara J; Augustine, Marilyn; Kahn, Leila; Martin, Emily; Oakley, Christine C; Carneiro, Raquel M; Tedesco, Mary Beth; Laslavic, Angela; Sereika, Susan M; Bisello, Alessandro; Garcia-Ocaña, Adolfo; Gundberg, Caren M; Cauley, Jane A; Stewart, Andrew F

    2013-01-01

    Parathyroid hormone-related protein (PTHrP)(1–36) increases lumbar spine (LS) bone mineral density (BMD), acting as an anabolic agent when injected intermittently, but has not been directly compared to parathyroid hormone (PTH)(1–34). We performed a three month, randomized, prospective study in 105 postmenopausal women with low bone density or osteoporosis comparing daily subcutaneous injections of PTHrP(1–36) to PTH(1–34). Thirty-five women were randomized to each of three groups: PTHrP(1–36) 400 μg/d; PTHrP(1–36) 600 μg/d; and PTH(1–34) 20 μg/d. The primary outcomes measures were changes in amino-terminal telopeptides of procollagen 1 (PINP) and carboxy-terminal telopeptides of collagen 1 (CTX). Secondary measures included safety parameters, 1,25(OH)2vitamin D and BMD. The increase in bone resorption (CTX) by PTH(1–34) (92%) (p<0.005) was greater than for PTHrP(1–36) (30%) (p<0.05). PTH(1–34) also increased bone formation (PINP) (171%) (p<0.0005) more than either dose of PTHrP(1–36) (46 & 87%). The increase in PINP was earlier (day 15) and greater than the increase in CTX for all three groups. LS BMD increased equivalently in each group (p<0.05 for all). Total hip (TH) and femoral neck (FN) BMD increased equivalently in each group but were only significant for the two doses of PTHrP(1–36) (p<0.05) at the TH, and for PTHrP(1–36) 400 (p<0.05) at the FN. PTHrP(1–36) 400 induced mild, transient (day 15) hypercalcemia. PTHrP(1–36) 600 required a dose reduction for hypercalcemia in three subjects. PTH(1–34) was not associated with hypercalcemia. Each peptide induced a marked biphasic increase in 1,25(OH)2D. Adverse events (AE) were similar among the three groups. This study demonstrates that PTHrP(1–36) and PTH(1–34) cause similar increases in LS BMD. PTHrP(1–36) also increased hip BMD. PTH(1–34) induced greater changes in bone turnover than PTHrP(1–36). PTHrP(1–36) was associated with mild transient hypercalcemia. Longer

  12. Rumen microorganisms, methane production, and microbial protein synthesis affected by mangosteen peel powder supplement in lactating dairy cows.

    PubMed

    Polyorach, Sineenart; Wanapat, Metha; Cherdthong, Anusorn; Kang, Sungchhang

    2016-03-01

    Four crossbred dairy cows (50 % Holstein-Friesian × 50 % Thai native), 404 ± 50.0 kg of body weight (4 years old) and 90 ± 5 day in milk with daily milk production of 9 ± 2.0 kg/day, were randomly assigned according to a 4 × 4 Latin square design to study the effect of mangosteen (Garcinia mangostana) peel powder (MSP) supplementation on rumen microorganisms, methane production, and microbial protein synthesis fed concentrate containing yeast fermented cassava chip protein (YEFECAP). The treatments were different levels of MSP supplementation at 0, 100, 200, and 300 g/head/day. Rice straw was used as a roughage source fed ad libitum, and concentrate containing YEFECAP at 200 g/kg concentrate was offered corresponding to concentrate-to-milk-yield ratio at 1:2. A quantitative real-time PCR approach was used to determine the population densities of ruminal microorganisms. The results revealed that supplementation of MSP did not affect on Fibrobactor succinogenes, Ruminococcus flavefaciens, and Ruminococcus albus (P > 0.05). However, total bacteria was linearly increased (P < 0.01) while methanogens and protozoal population were linearly decreased (P < 0.01) with increasing level of MSP supplementation. Increasing level of MSP supplement could decrease rumen methane production from 27.5 to 23.7 mmol/100 ml(3). Furthermore, cows that received MSP at 300 g/head/day had the highest microbial crude protein and efficiency of rumen microbial N synthesis (416.8 g/day and 16.2 g/kg organic matter truly digested in the rumen (OMDR), respectively). In conclusion, supplementation of MSP at 300 g/head/day with YEFECAP as a protein source in the concentrate mixture revealed an enhancement of rumen fermentation and methane reduction in lactating dairy cows.

  13. Lactation and body composition responses to fat and protein supplies during the dry period in under-conditioned dairy cows.

    PubMed

    Jaurena, G; Moorby, J M

    2017-02-01

    An experiment was designed to study the effect of precalving supplementation with protein (Pr) and rumen-inert fat (F) on body composition and subsequent milk production and composition. Forty Holstein-Friesian dairy cows were allocated to 1 of 4 dietary treatments in the dry period (DP) based on a first-cut ryegrass silage, with 6 mature (in their third or greater pregnancy) and 4 young (in their second pregnancy) cows per treatment. These were low Pr, low F (silage alone); low Pr, high F (silage with 10% rumen-inert fat, mixed on a dry matter basis); high Pr, low F [silage with 5% high-protein corn gluten meal (CGM)]; and high Pr, high F (silage with 5% CGM and 10% rumen-inert fat). All the diets were individually offered ad libitum and dry matter intake (DMI) was recorded daily during the DP. After calving, all cows received ryegrass silage plus 8 kg/d of a commercial dairy concentrate. During the DP, DMI was higher for mature than for young cows. All animals recovered body condition score (0.13 units/wk, 1-5 scale), reaching a maximum score of 2.4 some days before calving. Precalving maximum muscle longissimus dorsi (LD) depth was greater for mature (47.5 mm) than for young cows (45.7 mm), and milk fat concentration was also higher for mature than for young cows (40.2 and 39.0 g/kg, respectively). Supplementation with CGM increased maximum LD depth (from 45.9 to 47.6 mm), calf birth weight (low Pr = 43.2, high Pr = 46.3 kg), and milk crude protein concentration (from 30.8 to 31.6 g/kg). Fat supplementation in the DP of the mature cows increased maximum back fat depth (from 3.6 to 4.5 mm), milk yield (low F = 26.3, high F = 28.7 kg/d), and Pr yields (low F = 837, high F = 899 g/d). Inclusion of F in the DP diets reduced casein concentration in milk at wk 3 of lactation from 26.3 to 24.5 g/kg. Milk CP yield was also increased by CGM supplementation when compared within cows receiving F-supplemented silages (low Pr, high F = 832 g/d; high Pr, high F= 877 g/d). It

  14. Periosteal PTHrP regulates cortical bone modeling during linear growth in mice.

    PubMed

    Wang, Meina; VanHouten, Joshua N; Nasiri, Ali R; Tommasini, Steven M; Broadus, Arthur E

    2014-07-01

    The modeling of long bone surfaces during linear growth is a key developmental process, but its regulation is poorly understood. We report here that parathyroid hormone-related peptide (PTHrP) expressed in the fibrous layer of the periosteum (PO) drives the osteoclastic (OC) resorption that models the metaphyseal-diaphyseal junction (MDJ) in the proximal tibia and fibula during linear growth. PTHrP was conditionally deleted (cKO) in the PO via Scleraxis gene targeting (Scx-Cre). In the lateral tibia, cKO of PTHrP led to a failure of modeling, such that the normal concave MDJ was replaced by a mound-like deformity. This was accompanied by a failure to induce receptor activator of NF-kB ligand (RANKL) and a 75% reduction in OC number (P ≤ 0.001) on the cortical surface. The MDJ also displayed a curious threefold increase in endocortical osteoblast mineral apposition rate (P ≤ 0.001) and a thickened cortex, suggesting some form of coupling of endocortical bone formation to events on the PO surface. Because it fuses distally, the fibula is modeled only proximally and does so at an extraordinary rate, with an anteromedial cortex in CD-1 mice that was so moth-eaten that a clear PO surface could not be identified. The cKO fibula displayed a remarkable phenotype, with a misshapen club-like metaphysis and an enlargement in the 3D size of the entire bone, manifest as a 40-45% increase in the PO circumference at the MDJ (P ≤ 0.001) as well as the mid-diaphysis (P ≤ 0.001). These tibial and fibular phenotypes were reproduced in a Scx-Cre-driven RANKL cKO mouse. We conclude that PTHrP in the fibrous PO mediates the modeling of the MDJ of long bones during linear growth, and that in a highly susceptible system such as the fibula this surface modeling defines the size and shape of the entire bone.

  15. Effect of dietary energy source and level on nutrient digestibility, rumen microbial protein synthesis, and milk performance in lactating dairy cows.

    PubMed

    Zhou, X Q; Zhang, Y D; Zhao, M; Zhang, T; Zhu, D; Bu, D P; Wang, J Q

    2015-10-01

    This study was conducted to examine the effects of dietary energy source and level on intake, digestion, rumen microbial protein synthesis, and milk production in lactating dairy cows, using corn stover as a forage source. Eight multiparous Holstein cows, 4 of which were fitted with rumen cannulas, were evaluated in a replicated 4 × 4 Latin square design, with each period lasting 21 d. The cows were randomly assigned into 4 treatment groups: low-energy (LE) ground corn (GC), LE steam-flaked corn (SFC), high-energy (HE) GC, and HE SFC. Changes to ruminal energy degradation rates were induced by feeding the cows diets of either finely ground corn or SFC as components of diets with the same total energy level. Milk yield, milk protein content and yield, and milk lactose yield all increased in response to higher levels of dietary energy, whereas contents of milk fat and lactose were unaffected. Cows fed HE diets had a higher crude microbial protein yield and total-tract apparent digestibility than those receiving LE diets. Milk yield, milk protein yield, and microbial protein yield were also higher when SFC replaced GC as the main energy source for lactating cows fed LE diets. These results suggest that an increased dietary energy level and ruminal degradation rate are beneficial to milk protein production, which we suggest is due to increased yields of microbial proteins, when cows are fed corn stover as a dietary forage source.

  16. Multiparous cows categorized by milk protein concentration and energy-corrected milk yield during early lactation--metabolism, productivity and effect of a short-term feed restriction.

    PubMed

    Sigl, T; Gellrich, K; Meyer, H H D; Kaske, M; Wiedemann, S

    2013-04-01

    The objective of this experiment was to study milk productivity, metabolic adaptation and effect of a short-term feed restriction (FR) on key performance indicators during early lactation in cows classified according to energy-corrected milk (ECM) yield and milk protein concentration. Twenty-three multiparous Holstein-Friesian cows were categorized in four groups according to respective averaged values on Days 23-25 postpartum: high ECM yield and high protein concentration; low ECM yield and low protein concentration; high ECM yield and low protein concentration and low ECM yield and high protein concentration. Dry matter intake was reduced to 68.3% for three subsequent days. Our results showed that short-time FR in early lactation succeeded in enhancing energy deficit of cows in all groups. Milk fat, milk protein and lactose concentrations as well as milk fat yield were not influenced by FR. Several hepatic genes encoding for enzymes involved in catabolism of amino acids, β-oxidation, gluconeogenesis and ketogenesis as well as mRNA encoding for insulin receptor showed increased transcript abundances after FR, primarily in cows with high milk yield and low milk protein concentration.

  17. Diagnostic value of lactate, procalcitonin, ferritin, serum-C-reactive protein, and other biomarkers in bacterial and viral meningitis: A cross-sectional study.

    PubMed

    Sanaei Dashti, Anahita; Alizadeh, Shekoofan; Karimi, Abdullah; Khalifeh, Masoomeh; Shoja, Seyed Abdolmajid

    2017-09-01

    There are many difficulties distinguishing bacterial from viral meningitis that could be reasonably solved using biomarkers. The aim of this study was to evaluate lactate, procalcitonin (PCT), ferritin, serum-CRP (C-reactive protein), and other known biomarkers in differentiating bacterial meningitis from viral meningitis in children.All children aged 28 days to 14 years with suspected meningitis who were admitted to Mofid Children's Hospital, Tehran, between October 2012 and November 2013, were enrolled in this prospective cross-sectional study. Children were divided into 2 groups of bacterial and viral meningitis, based on the results of cerebrospinal fluid (CSF) culture, polymerase chain reaction, and cytochemical profile. Diagnostic values of CSF parameters (ferritin, PCT, absolute neutrophil count [ANC], white blood cell count, and lactate) and serum parameters (PCT, ferritin, CRP, and erythrocyte sedimentation rate [ESR]) were evaluated.Among 50 patients with meningitis, 12 were diagnosed with bacterial meningitis. Concentrations of all markers were significantly different between bacterial and viral meningitis, except for serum (P = .389) and CSF (P = .136) PCT. The best rates of area under the receiver operating characteristic (ROC) curve (AUC) were achieved by lactate (AUC = 0.923) and serum-CRP (AUC = 0.889). The best negative predictive values (NPV) for bacterial meningitis were attained by ANC (100%) and lactate (97.1%).The results of our study suggest that ferritin and PCT are not strong predictive biomarkers. A combination of low CSF lactate, ANC, ESR, and serum-CRP could reasonably rule out the bacterial meningitis.

  18. Procalcitonin, C-reactive protein and serum lactate dehydrogenase in the diagnosis of bacterial sepsis, SIRS and systemic candidiasis.

    PubMed

    Miglietta, Fabio; Faneschi, Maria Letizia; Lobreglio, Giambattista; Palumbo, Claudio; Rizzo, Adriana; Cucurachi, Marco; Portaccio, Gerolamo; Guerra, Francesco; Pizzolante, Maria

    2015-09-01

    The aim of this study was to evaluate procalcitonin (PCT), C-reactive protein (CRP), platelet count (PLT) and serum lactate dehydrogenase (LDH) as early markers for diagnosis of SIRS, bacterial sepsis and systemic candidiasis in intensive care unit (ICU) patients. Based on blood culture results, the patients were divided into a sepsis group (70 patients), a SIRS group (42 patients) and a systemic candidiasis group (33 patients). PCT, CRP, LDH and PLT levels were measured on day 0 and on day 2 from the sepsis symptom onset. PCT levels were higher in Gram negative sepsis than those in Gram positive sepsis, although the P value between the two subgroups is not significant (P=0.095). Bacterial sepsis group had higher PCT and CRP levels compared with the systemic candidiasis group, whereas PLT and LDH levels showed similar levels in these two subgroups. The AUC for PCT (AUC: 0.892, P <0.001) was larger than for CRP (AUC: 0.738, P <0.001). The best cut-off values for PCT and CRP were 0.99 ng/mL and 76.2 mg/L, respectively. Diagnostic sensitivity and specificity for PCT were 84.3% and 81.8% whereas CRP showed a sensitivity of 77.2% and a specificity of 63.6%. However, PCT was unable to discriminate between SIRS and systemic candidiasis groups (P=0.093 N.S.). In conclusion, PCT can be used as a preliminary marker in the event of clinical suspicion of systemic candidiasis; however, low PCT levels (<0.99 ng/mL) necessarily require the use of other specific markers of candidaemia to confirm the diagnosis, due to great uniformity of PCT levels in systemic candidiasis and SIRS groups.

  19. Strong associations among rumen endotoxin and acute phase proteins with plasma minerals in lactating cows fed graded amounts of concentrate.

    PubMed

    Zebeli, Q; Dunn, S M; Ametaj, B N

    2010-04-01

    The objective of this investigation was to determine associations among rumen endotoxin, plasma serum amyloid A (SAA), and C-reactive protein (CRP) with plasma Ca, Fe, Zn, and Cu in lactating cows challenged with graded amounts of rolled barley grain in the diet (i.e., 0, 15, 30, and 45% of DMI). Correlative relationships among variables were determined by linear and nonlinear regression procedures adjusted for the effects of day, animal, and experimental period. Increasing the amount of grain in the diet was successful in inducing an acute phase response, as assessed by augmentation of rumen endotoxin and plasma CRP and SAA (P < 0.01). The correlative analysis revealed inverse, nonlinear relationships of rumen endotoxin and plasma SAA with circulating Ca. Interestingly, plasma Ca reached the asymptotic plateau at 10.6 mg/dL. The increase in rumen endotoxin was associated with an abrupt decrease in plasma Fe (R(2) = 0.91; P < 0.001). A similar relationship, although at a reduced estimation accuracy (R(2) = 0.21; P < 0.01), was observed between rumen endotoxin and plasma Zn. Augmentation of rumen endotoxin and plasma CRP resulted in a positive, biphasic response of plasma Cu. In conclusion, the increase in rumen endotoxin in response to high-grain diets, and the resulting increases in plasma SAA and CRP, were strongly correlated with fluctuations of plasma minerals. Results suggest that new feeding strategies should be developed to curb the release of endotoxin in the rumen fluid to prevent perturbing minerals in the plasma.

  20. Response of lactating dairy cows to steam-flaked sorghum, steam-flaked corn, or steam-rolled corn and protein sources of differing degradability.

    PubMed

    Santos, J E; Huber, J T; Theurer, C B; Nussio, L G; Tarazon, M; Santos, F A

    1999-04-01

    Protein sources with different degradabilities were fed to 48 lactating Holstein cows receiving 37 or 39% of dietary dry matter as steam-flaked sorghum (360 g/L), steam-flaked corn (360 g/L), or steam-rolled corn (490 g/L) in a 3 x 2 factorial arrangement of treatments. Cows were fed an alfalfa-based diet with 7% soybean meal or 5% of an animal-marine protein blend and 37 or 39% grain. Although not significant, cows fed flaked grain yielded a mean of 1.5 kg/d more milk than did those fed rolled grain. Gross feed efficiency was not affected by grain processing or protein source, but diets with the animal-marine protein blend had 9% higher estimated net energy for lactation than did diets with soybean meal. The greater gains in body weight and increased digestibility of the diets with the animal-marine protein blend verify the higher energy concentration of those diets. Yield of milk protein was increased by flaked grain or the animal-marine protein blend, and flaked grain increased percentages of lactose and solids nonfat. Increasing the ratio of rumen-degradable starch to rumen-degradable protein increased milk protein content and yield linearly and increased contents of lactose and solids nonfat. A linear response of dry matter, organic matter, crude protein, and starch digestibilities was observed as the ratio of rumen-degradable starch to rumen-degradable protein increased. These data show improved performance of dairy cows fed a high rumen-undegradable protein source with diets high in rumen-degradable starch from steam-processed grains.

  1. Lactate dehydrogenase-elevating virus

    USDA-ARS?s Scientific Manuscript database

    This book chapter describes the taxonomic classification of Lactate dehydrogenase-elevating virus (LDV). Included are: host, genome, classification, morphology, physicochemical and physical properties, nucleic acid, proteins, lipids, carbohydrates, geographic range, phylogenetic properties, biologic...

  2. Immunochemical detection of parathyroid hormone-related protein in the saccus vasculosus of a teleost fish.

    PubMed

    Devlin, A J; Danks, J A; Faulkner, M K; Power, D M; Canario, A V; Martin, T J; Ingleton, P M

    1996-01-01

    Using antisera to regions of human parathyroid hormone-related protein (PTHrP) the saccus vasculosus (SV) of the sea bream (Sparus aurata) has been shown to contain immunoreactive PTHrP. By immunohistochemistry (IHC) the epithelial coronet cells in fixed and wax-embedded SV tissue reacted with antisera to the prepro region of human PTHrP (-13 to +2), the N-terminus PTHrP (1-16), and the midmolecule PTHrP (50-69). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of saccus extracts and incubation media contained two major proteins of 14.3 and 15 kDa. By Western blotting these two proteins both reacted with the three antisera used for IHC, suggesting that they are immunochemically similar to human PTHrP (1-84). Ultrastructurally the coronet cells of Sparus saccus vasculosus resembled coronet cells described for other teleosts, with an abundant smooth endoplasmic reticulum (SER) which was more highly organized in the coronets. IHC at EM level showed reaction mainly with the membranes of the SER. These results suggest that S. aurata saccus vasculosus may produce a PTHrP-like molecule similar to human PTHrP.

  3. mTORC1 regulates PTHrP to coordinate chondrocyte growth, proliferation and differentiation

    PubMed Central

    Yan, Bo; Zhang, Zhongmin; Jin, Dadi; Cai, Chen; Jia, Chunhong; Liu, Wen; Wang, Ting; Li, Shengfa; Zhang, Haiyan; Huang, Bin; Lai, Pinglin; Wang, Hua; Liu, Anling; Zeng, Chun; Cai, Daozhang; Jiang, Yu; Bai, Xiaochun

    2016-01-01

    Precise coordination of cell growth, proliferation and differentiation is essential for the development of multicellular organisms. Here, we report that although the mechanistic target of rapamycin complex 1 (mTORC1) activity is required for chondrocyte growth and proliferation, its inactivation is essential for chondrocyte differentiation. Hyperactivation of mTORC1 via TSC1 gene deletion in chondrocytes causes uncoupling of the normal proliferation and differentiation programme within the growth plate, resulting in uncontrolled cell proliferation, and blockage of differentiation and chondrodysplasia in mice. Rapamycin promotes chondrocyte differentiation and restores these defects in mutant mice. Mechanistically, mTORC1 downstream kinase S6K1 interacts with and phosphorylates Gli2, and releases Gli2 from SuFu binding, resulting in nuclear translocation of Gli2 and transcription of parathyroid hormone-related peptide (PTHrP), a key regulator of bone development. Our findings demonstrate that dynamically controlled mTORC1 activity is crucial to coordinate chondrocyte proliferation and differentiation partially through regulating Gli2/PTHrP during endochondral bone development. PMID:27039827

  4. Variance Components and Genetic Parameters Estimated for Fat and Protein Content in Individual Months of Lactation: The Case of Tsigai Sheep.

    PubMed

    Oravcová, Marta

    2016-02-01

    The objective of this study was to assess variance components and genetic parameters for fat and protein content in Tsigai sheep using multivariate animal models in which fat and protein content in individual months of lactation were treated as different traits, and univariate models in which fat and protein content were treated as repeated measures of the same traits. Test day measurements were taken between the second and the seventh month of lactation. The fixed effects were lactation number, litter size and days in milk. The random effects were animal genetic effect and permanent environmental effect of ewe. The effect of flock-year-month of test day measurement was fitted either as a fixed (FYM) or random (fym) effect. Heritabilities for fat content were estimated between 0.06 and 0.17 (FYM fitted) and between 0.06 and 0.11 (fym fitted). Heritabilities for protein content were estimated between 0.15 and 0.23 (FYM fitted) and between 0.10 and 0.18 (fym fitted). For fat content, variance ratios of permanent environmental effect of ewe were estimated between 0.04 and 0.11 (FYM fitted) and between 0.02 and 0.06 (fym fitted). For protein content, variance ratios of permanent environmental effect of ewe were estimated between 0.13 and 0.20 (FYM fitted) and between 0.08 and 0.12 (fym fitted). The proportion of phenotypic variance explained by fym effect ranged from 0.39 to 0.43 for fat content and from 0.25 to 0.36 for protein content. Genetic correlations between individual months of lactation ranged from 0.74 to 0.99 (fat content) and from 0.64 to 0.99 (protein content). Fat content heritabilities estimated with univariate animal models roughly corresponded with heritability estimates from multivariate models: 0.13 (FYM fitted) and 0.07 (fym fitted). Protein content heritabilities estimated with univariate animal models also corresponded with heritability estimates from multivariate models: 0.18 (FYM fitted) and 0.13 (fym fitted).

  5. Low degradable protein supply to increase nitrogen efficiency in lactating dairy cows and reduce environmental impacts at barn level.

    PubMed

    Edouard, N; Hassouna, M; Robin, P; Faverdin, P

    2016-02-01

    Generally, <30% of dairy cattle's nitrogen intake is retained in milk. Large amounts of nitrogen are excreted in manure, especially in urine, with damaging impacts on the environment. This study explores the effect of lowering dietary degradable nitrogen supplies--while maintaining metabolisable protein--on dairy cows' performance, nitrogen use efficiency and gas emissions (NH3, N2O, CH4) at barn level with tied animals. Two dietary N concentrations (CP: 12% DM for LowN; 18% DM for HighN) were offered to two groups of three lactating dairy cows in a split-plot design over four periods of 2 weeks. Diets were formulated to provide similar metabolisable protein supply, with degradable N either in deficit or in excess (PDIN of 84 and 114 g/kg DM for LowN and HighN, respectively). Cows ingested 0.8 kg DM/day less on the LowN diet, which was also 2.5% less digestible. Milk yield and composition were not significantly affected. N exported in milk was 5% lower (LowN: 129 g N/day; HighN: 136 g N/day; P<0.001) but milk protein yield was not significantly affected (LowN: 801 g/day; HighN: 823 g/day; P=0.10). Cows logically ingested less nitrogen on the LowN diet (LowN: 415 g N/day; HighN: 626 g N/day; P<0.001) resulting in a higher N use efficiency (N milk/N intake; LowN: 0.31; HighN: 0.22; P<0.001). N excreted in urine was almost four times lower on the LowN diet (LowN: 65 g N/day; HighN: 243 g N/day; P<0.001) while urinary urea N concentration was eightfold lower (LowN: 4.6 g/l; HighN: 22.9 g/l; P<0.001). Ammonia emission (expressed in g/h in order to remove periods of the day with potential interferences with volatile molecules from feed) was also lower on the LowN diet (LowN: 1.03 g/h per cow; HighN: 1.25 g/h per cow; P<0.05). Greenhouse gas emissions (N2O and CH4) at barn level were not significantly affected by the amount of dietary N. Offering low amounts of degradable protein with suitable metabolisable protein amounts to cattle improved nitrogen use efficiency and

  6. Parathyroid hormone-related protein overexpression protects goat mammary gland epithelial cells from calcium-sensing receptor activation-induced apoptosis.

    PubMed

    Li, Hui; Sun, Yongsen; Zheng, Huiling; Li, Lihui; Yu, Qian; Yao, Xiaotong

    2015-01-01

    Normal mammary gland epithelial cells and breast cancer cells express the calcium-sensing receptor (CaSR), which is the master regulator of systemic calcium metabolism. During lactation, activation of the CaSR in mammary epithelial cells downregulates parathyroid hormone-related protein (PTHrP) levels in milk and in the circulation, and increases calcium transport into milk. However, very little information is available on the role of CaSR in goat mammary gland epithelial cells (GMECs) apoptosis. In this investigation, the full-length cDNA of CaSR from Xinong Saanen dairy goats was cloned, which contains an open-reading frame of 3,258 bp encoding 1,085 amino acids with a predicted molecular weight of 121.0 kDa and an isoelectric point of 5.65. The amino acid sequence is highly homologous with sheep, and the goat CaSR gene is mapped to chromosome 1. Quantitative real-time PCR suggested that CaSR was predominantly expressed in the heart, kidney and mammary gland. Then, we found the stimulation of CaSR with its activator gadolinium chloride (GdCl3) contributed to increase CaSR mRNA levels in GMECs and simultaneously promoted cell apoptosis, and these effects were abrogated partially by NPS2390 which is an inhibitor of CaSR. We also demonstrated that Ca(2+) increased CaSR mRNA levels and induced GMECs apoptosis and restrained cell proliferation. In contrast, PTHrP overexpression protected GMECs from calcium-induced apoptosis, and promoted cell proliferation. In conclusion, these results suggest that PTHrP overexpression protects GMECs from CaSR activation-induced apoptosis.

  7. Effects of protein level, methionine supplementation and carbohydrate type of the diet on liver lipid and plasma free threonine contents in the lactating rat.

    PubMed

    Leclerc, J; Chanussot, B; Miller, M L; Poisson, J P; Belleville, J

    1989-01-01

    Eight groups of 13-15 female rats were fed purified diets after littering. Four groups received a low protein (8% casein) diet (groups 8) and the others, a normal protein (20% casein) diet (groups 20). Carbohydrates were supplied either as starch (groups S) or as starch plus 40% fructose (groups F). Half the animals received a 0.4% methionine supplementation (groups M). Four or five dams per group were sacrificed on days 2, 7 and 14 after littering. The diet intake was increased by methionine supplementation, substitution of starch for fructose and increased protein content, mainly during the second week of lactation. This influenced weight variation of the dams and litter growth. On all days, the plasma levels of cholesterol esters, triglycerides and phospholipids were positively correlated with the dietary protein level. On days 7 and 14, the liver neutral lipid content was increased in rats fed the low protein diets supplemented with methionine (groups 8SM and 8FM) and the normal protein diets containing 40% fructose (groups 20F and 20FM). The plasma free threonine content was positively correlated with the protein level in the diet. On day 14, rats fed a low protein diet had a threonine deficiency, except those in groups 8S and 8F. The plasma free threonine content of these rats was not reduced, possibly due to an impaired utilization of this amino acid. The liver lipidosis observed during lactation, in contrast to that observed during growth with a low protein diet, was not due to a threonine deficiency.

  8. Perinatal protein restriction affects milk free amino acid and fatty acid profile in lactating rats: potential role on pup growth and metabolic status.

    PubMed

    Martin Agnoux, Aurore; Antignac, Jean-Philippe; Boquien, Clair-Yves; David, Agnes; Desnots, Emmanuelle; Ferchaud-Roucher, Veronique; Darmaun, Dominique; Parnet, Patricia; Alexandre-Gouabau, Marie-Cécile

    2015-07-01

    Perinatal undernutrition affects not only fetal and neonatal growth but also adult health outcome, as suggested by the metabolic imprinting concept. Although maternal milk is the only channel through which nutrients are transferred from mother to offspring during the postnatal period, the impact of maternal undernutrition on milk composition is poorly understood. The present study investigates, in a rat model of nutritional programming, the effects of feeding an isocaloric, low-protein diet throughout gestation and lactation on milk composition and its possible consequences on offspring's growth and metabolic status. We used an integrated methodological approach that combined targeted analyses of macronutrients, free amino acid and fatty acid content throughout lactation, with an untargeted mass-spectrometric-based metabolomic phenotyping. Whereas perinatal dietary protein restriction failed to alter milk protein content, it dramatically decreased the concentration of most free amino acids at the end of lactation. Interestingly, a decrease of several amino acids involved in insulin secretion or gluconeogenesis was observed, suggesting that maternal protein restriction during the perinatal period may impact the insulinotrophic effect of milk, which may, in turn, account for the slower growth of the suckled male offspring. Besides, the decrease in sulfur amino acids may alter redox status in the offspring. Maternal undernutrition was also associated with an increase in milk total fatty acid content, with modifications in their pattern. Altogether, our results show that milk composition is clearly influenced by maternal diet and suggest that alterations in milk composition may play a role in offspring growth and metabolic programming. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Corn steep liquor and fermented ammoniated condensed whey as protein sources for lactating cows and yearling heifers grazing winter native range

    SciTech Connect

    Wagner, J.J.; Lusby, K.S.; Horn, G.W.; Dvorak, M.J.

    1982-06-01

    Corn steep liquor (CSL) and fermented ammoniated condensed whey (FACW) were compared to cottonseed meal (CSM) as protein sources for wintering 61 lactating first-calf Hereford heifers and 32 yearling Hereford heifers on native range. Cattle were allotted by weight and individually fed 6 days per week for 12 weeks one of four protein treatments: negative control (NC), positive control (PC), CSL and FACW to provide .7, 1.5, .15 and 1.5 lb crude protein (CP) per day, respectively, to the lacating heifers and .2, .4, .4 and .4lb cP per day, respectively, to the yearling heifers. CMS was supplied in the CSL and FACW treatments at the same level as in the negative control. Lactating heifers fed the NC lost more (P less than .005) weight and body condition (120 lb and 1.6 units) than those fed the PC (45.8 lb and .9 units). Weight and condition losses were similar (P more than .05) for lactating heifers fed PC, CSL and FACW. Yearling heifers fed the NC lost more (P less than .005) weight than those fed the PC (49.4 vs 10.6 lb). Yearling heifers fed CSL and FACW gained more (P less than .005) weight than those fed the PC (17.6 and 9.3 vs - 10.6 lb). Feeding CSL resulted in signficantly lower rumen pH, lower ruminal acetate and higher ruminal butyrate, isovalerate and caproate levels than did feeding either control. Supplementing with FACW produced significantly lower rumen pH, higher rumen ammonia and soluble carbohydrate levels, lower ruminal acetate, and higher ruminal propionate and butyrate concentrations than did either control supplement. Corn steep liquor and FDCW appear to be effective protein sources for cows and heifers grazing winter native range.

  10. Evaluation of urinary S100B protein level and lactate/creatinine ratio for early diagnosis and prognostic prediction of neonatal hypoxic-ischemic encephalopathy.

    PubMed

    Liu, Li; Zheng, Chong-Xun; Peng, Shu-Feng; Zhou, Hong-Yan; Su, Zu-You; He, Li; Ai, Ting

    2010-01-01

    Early identification and prevention of hypoxic-ischemic encephalopathy (HIE) in newborns may reduce neonatal mortality and neurological dysfunction. To analyze the diagnostic and prognostic values of urinary S100B level and lactate/creatinine ratio in newborns with HIE. Seventy-eight full-term newborns with HIE and 25 normal newborns were enrolled. The Neonatal Behavioral Neurological Assessment (NBNA) and Developmental Screening Test were scored. The concentration of urinary S100B protein was determined using the S100B enzyme-linked immunosorbent assay and the levels of urinary lactate and creatinine were measured with the enzyme colorimetric method. Urinary S100B level on days 1-3 after birth and lactate/creatinine ratio on day 1 were significantly higher in newborns with HIE than those in the control group. Both indexes were positively correlated with the clinical grading of HIE. A cutoff value for the S100B level of 0.47 microg/l on day 3 after birth had a sensitivity of 90% and specificity of 92% for prediction of HIE. A lactate/creatinine ratio of more than 0.55 on day 1 showed the highest sensitivity (92%) and specificity (90%). A combination of both indexes improved the sensitivity and specificity to 99 and 97%, respectively. A negative correlation of both lactate/creatinine ratio on day 1 and S100B level on days 1-3 after birth with the NBNA score was identified on days 3, 7 and 14 after birth. The Developmental Screening Test score of 36 newborns with HIE within 6 months after birth showed that 65% of infants with moderate and high HIE had an abnormal developmental quotient. These data suggest that early measurement of both S100B level and lactate/creatinine ratio in the urine of newborns with HIE is a practical convenient and sensitive way to improve diagnosis on the third day of life and prognostic prediction of HIE. Copyright 2009 S. Karger AG, Basel.

  11. Feeding rumen-protected methionine pre- and postpartum increases milk protein content and yield in early lactation

    USDA-ARS?s Scientific Manuscript database

    Objectives were to evaluate the effects of feeding rumen-protected methionine (MET) from 23 d (±12) before calving until 98 days in milk (DIM) on lactation performance, dry matter intake (DMI), body condition score (BCS) and body weight (BW) change of dairy cows. Multiparous Holstein cows (n = 223) ...

  12. Effects of rumen-undegradable protein sources and supplemental 2-hydroxy-4-(methylthio)-butanoic acid and lysine-HCl on lactation performance in dairy cows.

    PubMed

    Johnson-VanWieringen, L M; Harrison, J H; Davidson, D; Swift, M L; von Keyserlingk, M A G; Vazquez-Anon, M; Wright, D; Chalupa, W

    2007-11-01

    One hundred primiparous and multiparous Holstein cows were used in an experiment to evaluate the effect of supplementing diets with either a plant- or an animal-based source of rumen-undegradable protein (RUP), with or without AA supplementation, during the transition period and early lactation on milk production response. The experimental design was a randomized block design with approximately one-third of the cows being primiparous. Cows were assigned to 1 of 4 prepartum diets introduced 3 wk before the expected calving date and switched to the corresponding postpartum diet at calving. Diets 1 (AMI) and 2 (AMI+) included a vegetable RUP source (heat- and lignosulfonate-treated canola meal), with diet 2 containing supplemental Lys x HCl and Met hydroxy analog sources [D,L-2 hydroxy-4-(methylthio)-butanoic acid; Alimet feed supplement]. Diets 3 (PRO) and 4 (PRO+) consisted of a blend of animal RUP sources (blood meal, fish meal, feather meal, and porcine meat and bone meal), with diet 4 containing supplemental Lys x HCl and Met hydroxy analog sources [D,L-2 hydroxy-4-(methylthio)-butanoic acid; Alimet]. During the first 4 wk of lactation, dry matter intake was less when synthetic Lys x HCl and Alimet were supplemented, but this effect was no longer evident in wk 5 to 9 of the experiment. Interestingly, despite the initial decrease in dry matter intake in the cows fed AA-supplemented diets, there was no effect of treatment on milk production or the ratio of fat-corrected milk to dry matter intake throughout the 17 wk of the study. Undegradable protein source (vegetable vs. animal) did not affect dry matter intake, milk production, or 3.5% fat-corrected milk production for the first 17 wk of lactation. The results of this study indicate that heat- and lignosulfonate-treated canola meal can be used as a source of undegradable protein in place of high-quality rumen-undegradable animal protein sources without negative effects on milk production when diets are equivalent

  13. Hippocampal mechanisms in impaired spatial learning and memory in male offspring of rats fed a low-protein isocaloric diet in pregnancy and/or lactation.

    PubMed

    Reyes-Castro, L A; Padilla-Gómez, E; Parga-Martínez, N J; Castro-Rodríguez, D C; Quirarte, G L; Díaz-Cintra, S; Nathanielsz, P W; Zambrano, E

    2017-08-26

    Maternal nutritional challenges during fetal and neonatal development result in developmental programming of multiple offspring organ systems including brain maturation and function. A maternal low-protein diet during pregnancy and lactation impairs associative learning and motivation. We evaluated effects of a maternal low-protein diet during gestation and/or lactation on male offspring spatial learning and hippocampal neural structure. Control mothers (C) ate 20% casein and restricted mothers (R) 10% casein, providing four groups: CC, RR, CR, and RC (first letter pregnancy, second lactation diet). We evaluated the behavior of young adult male offspring around postnatal day 110. Corticosterone and ACTH were measured. Males were tested for 2 days in the Morris water maze (MWM). Stratum lucidum mossy fiber (MF) area, total and spine type in basal dendrites of stratum oriens in the hippocampal CA3 field were measured. Corticosterone and ACTH were higher in RR vs. CC. In the MWM acquisition test CC offspring required two, RC three, and CR seven sessions to learn the maze. RR did not learn in eight trials. In a retention test 24 h later, RR, CR, and RC spent more time locating the platform and performed fewer target zone entries than CC. RR and RC offspring spent less time in the target zone than CC. MF area, total, and thin spines were lower in RR, CR, and RC than CC. Mushroom spines were lower in RR and RC than CC. Stubby spines were higher in RR, CR, and RC than CC. We conclude that maternal low-protein diet impairs spatial acquisition and memory retention in male offspring, and that alterations in hippocampal presynaptic (MF), postsynaptic (spines) elements and higher glucocorticoid levels are potential mechanisms to explain these learning and memory deficits. © 2017 Wiley Periodicals, Inc.

  14. Nutritional Recovery with a Soybean Diet after Weaning Reduces Lipogenesis but Induces Inflammation in the Liver in Adult Rats Exposed to Protein Restriction during Intrauterine Life and Lactation

    PubMed Central

    Reis, Sílvia Regina de Lima; Feres, Naoel Hassan; Ignacio-Souza, Leticia Martins; Veloso, Roberto Vilela; Arantes, Vanessa Cristina; Kawashita, Nair Honda; Colodel, Edson Moleta; Botosso, Bárbara Laet; Reis, Marise Auxiliadora de Barros; Latorraca, Márcia Queiroz

    2015-01-01

    We evaluated the effects of postweaning nutritional recovery with a soybean flour diet on de novo hepatic lipogenesis and inflammation in adult rats exposed to protein restriction during intrauterine life and lactation. Rats from mothers fed with protein (casein) in a percentage of 17% (control, C) or 6% (low, L) during pregnancy and lactation were fed with diet that contained 17% casein (CC and LC groups, resp.) or soybean (CS and LS groups, resp.) after weaning until 90 days of age. LS and CS rats had low body weight, normal basal serum triglyceride levels, increased ALT concentrations, and high HOMA-IR indices compared with LC and CC rats. The soybean diet reduced PPARγ as well as malic enzyme and citrate lyase contents and activities. The lipogenesis rate and liver fat content were lower in LS and CS rats relative to LC and CC rats. TNFα mRNA and protein levels were higher in LS and CS rats than in LC and CC rats. NF-κB mRNA levels were lower in the LC and LS groups compared with the CC and LC groups. Thus, the soybean diet prevented hepatic steatosis at least in part through reduced lipogenesis but resulted in TNFα-mediated inflammation. PMID:25892856

  15. Cell-cell and intracellular lactate shuttles.

    PubMed

    Brooks, George A

    2009-12-01

    Once thought to be the consequence of oxygen lack in contracting skeletal muscle, the glycolytic product lactate is formed and utilized continuously in diverse cells under fully aerobic conditions. 'Cell-cell' and 'intracellular lactate shuttle' concepts describe the roles of lactate in delivery of oxidative and gluconeogenic substrates as well as in cell signalling. Examples of the cell-cell shuttles include lactate exchanges between between white-glycolytic and red-oxidative fibres within a working muscle bed, and between working skeletal muscle and heart, brain, liver and kidneys. Examples of intracellular lactate shuttles include lactate uptake by mitochondria and pyruvate for lactate exchange in peroxisomes. Lactate for pyruvate exchanges affect cell redox state, and by itself lactate is a ROS generator. In vivo, lactate is a preferred substrate and high blood lactate levels down-regulate the use of glucose and free fatty acids (FFA). As well, lactate binding may affect metabolic regulation, for instance binding to G-protein receptors in adipocytes inhibiting lipolysis, and thus decreasing plasma FFA availability. In vitro lactate accumulation upregulates expression of MCT1 and genes coding for other components of the mitochondrial reticulum in skeletal muscle. The mitochondrial reticulum in muscle and mitochondrial networks in other aerobic tissues function to establish concentration and proton gradients necessary for cells with high mitochondrial densities to oxidize lactate. The presence of lactate shuttles gives rise to the realization that glycolytic and oxidative pathways should be viewed as linked, as opposed to alternative, processes, because lactate, the product of one pathway, is the substrate for the other.

  16. Cell–cell and intracellular lactate shuttles

    PubMed Central

    Brooks, George A

    2009-01-01

    Once thought to be the consequence of oxygen lack in contracting skeletal muscle, the glycolytic product lactate is formed and utilized continuously in diverse cells under fully aerobic conditions. ‘Cell–cell’ and ‘intracellular lactate shuttle’ concepts describe the roles of lactate in delivery of oxidative and gluconeogenic substrates as well as in cell signalling. Examples of the cell–cell shuttles include lactate exchanges between between white-glycolytic and red-oxidative fibres within a working muscle bed, and between working skeletal muscle and heart, brain, liver and kidneys. Examples of intracellular lactate shuttles include lactate uptake by mitochondria and pyruvate for lactate exchange in peroxisomes. Lactate for pyruvate exchanges affect cell redox state, and by itself lactate is a ROS generator. In vivo, lactate is a preferred substrate and high blood lactate levels down-regulate the use of glucose and free fatty acids (FFA). As well, lactate binding may affect metabolic regulation, for instance binding to G-protein receptors in adipocytes inhibiting lipolysis, and thus decreasing plasma FFA availability. In vitro lactate accumulation upregulates expression of MCT1 and genes coding for other components of the mitochondrial reticulum in skeletal muscle. The mitochondrial reticulum in muscle and mitochondrial networks in other aerobic tissues function to establish concentration and proton gradients necessary for cells with high mitochondrial densities to oxidize lactate. The presence of lactate shuttles gives rise to the realization that glycolytic and oxidative pathways should be viewed as linked, as opposed to alternative, processes, because lactate, the product of one pathway, is the substrate for the other. PMID:19805739

  17. Effect of dietary starch level and high rumen-undegradable protein on endocrine-metabolic status, milk yield, and milk composition in dairy cows during early and late lactation.

    PubMed

    Piccioli-Cappelli, F; Loor, J J; Seal, C J; Minuti, A; Trevisi, E

    2014-12-01

    Diet composition defines the amount and type of nutrients absorbed by dairy cows. Endocrine-metabolic interactions can influence these parameters, and so nutrient availability for the mammary gland can significantly vary and affect milk yield and its composition. Six dairy cows in early and then late lactation received, for 28 d in a changeover design, 2 diets designed to provide, within the same stage of lactation, similar amounts of rumen fermentable material but either high starch plus sugar (HS) content or low starch plus sugar content (LS). All diets had similar dietary crude protein and calculated supply of essential amino acids. Dry matter intake within each stage of lactation was similar between groups. Milk yield was similar between groups in early lactation, whereas a higher milk yield was observed in late lactation when feeding HS. At the metabolic level, the main difference observed between the diets in both stages of lactation was lower blood glucose in cows fed LS. The lower glucose availability during consumption of LS caused substantial modifications in the circulating and postprandial pattern of metabolic hormones. Feeding LS versus HS resulted in an increase in the ratio of bovine somatotropin to insulin. This increased mobilization of lipid reserves resulted in higher blood concentrations of nonesterified fatty acids and β-hydroxybutyrate, which contributed to the higher milk fat content in both stages of lactation in the LS group. This greater recourse to body fat stores was confirmed by the greater loss of body weight during early lactation and the slower recovery of body weight in late lactation in cows fed LS. The lower insulin to glucagon ratio observed in cows fed LS in early and late lactation likely caused an increase in hepatic uptake and catabolism of amino acids, as confirmed by the higher blood urea concentrations. Despite the higher catabolism of amino acids in LS in early lactation, similar milk protein output was observed for both

  18. Effect of replacing soybean meal protein with protein from upland cottonseed, Pima cottonseed, or extruded Pima cottonseed on production of lactating dairy cows.

    PubMed

    Broderick, G A; Kerkman, T M; Sullivan, H M; Dowd, M K; Funk, P A

    2013-04-01

    Pima cotton production is increasing in the United States, but Pima cottonseed generally contains higher concentrations of the antinutritive pigment gossypol than conventional upland cottonseed. Heating promotes the reaction of gossypol with protein, reducing gossypol absorption and toxicity. The objective of this study was to assess the nutritional value for dairy cattle of Pima cottonseed cake (PCSC) that was heated and oil largely removed by an experimental extrusion process, compared with upland cottonseed (UCS) and Pima cottonseed (PCS). The PCS had greater crude protein (CP) and ether extract, less neutral detergent fiber (NDF) and acid detergent fiber (ADF), similar total gossypol, but higher (-)-gossypol isomer compared with UCS. Extrusion reduced lipid content by 73%, increased concentrations of CP, NDF, and ADF, and reduced total gossypol, (+)-gossypol, and (-)-gossypol in PCSC versus PCS. Forty lactating Holsteins (8 with ruminal cannulas) were blocked by days in milk into 5 squares in a replicated, incomplete 8 × 8 Latin square, and were fed diets containing, on a dry matter (DM) basis, 30% alfalfa silage, 31% corn silage, 21 to 25% high-moisture corn, and about 15% CP. Diets were fed as total mixed rations for ad libitum intake. Supplemental CP was from (1) solvent soybean meal (SSBM) only or 50% from SSBM plus 50% from (2) UCS, (3) PCS, (4) PCSC, (5) UCS plus PCS, and (6) UCS plus PCSC, or (7) 50% from expeller soybean meal (ESBM) plus 50% from PCS, and (8) 50% from ESBM plus 50% from PCSC. Periods were 4 wk long (total of 16 wk); production data were collected over the last 2 wk and blood and ruminal samples were taken on d 28 of each period. Data were analyzed using Proc Mixed of SAS (SAS Institute Inc., Cary, NC). Diet affected dry matter intake, with greatest intake on diet 6 and lowest intake on diets 1 and 3. The highest milk fat content was observed on diet 5 and the greatest fat yield on diet 7; fat content and yield were lowest on diet 1

  19. Feeding glycerol-enriched yeast culture improves performance, energy status, and heat shock protein gene expression of lactating Holstein cows under heat stress.

    PubMed

    Liu, J; Ye, G; Zhou, Y; Liu, Y; Zhao, L; Liu, Y; Chen, X; Huang, D; Liao, S F; Huang, K

    2014-06-01

    This study was conducted to evaluate the effects of supplemental common yeast culture (CY) and glycerol-enriched yeast culture (GY) on performance, plasma metabolites, antioxidant status, and heat shock protein 70 (HSP70) mRNA expression in lactating Holstein cows under heat stress. During summer months, 30 healthy multiparous lactating cows (parity 3.25 ± 0.48; 60 ± 13 d in milk [DIM]; 648 ± 57 kg BW; an average milk yield of 33.8 ± 1.6 kg/d) were blocked by parity, previous milk yield, and DIM and randomly allocated to 3 dietary treatments: no supplemental yeast culture (Control), 1 L/d of CY (33.1 g yeast) per cow, and 2 L/d of GY (153.2 g glycerol and 31.6 g yeast) per cow. During the 60-d experiment, values of air temperature and relative humidity inside the barn were recorded hourly every 3 d to calculate temperature-humidity index (THI). Weekly rectal temperatures (RT) and respiration rates and daily DMI and milk yield were recorded for all cows. Milk and blood samples were taken twice monthly, and BW and BCS were obtained on d 0 and 60. In this experiment, THI values indicated cows experienced a moderate heat stress. Cows supplemented with CY and GY had greater yields of milk, energy-corrected milk and milk fat, and milk fat percent but lower HSP70 mRNA expression in peripheral blood lymphocytes than Control cows (P < 0.05). Supplementing CY and GY tended (P < 0.15) to decrease RT at 1400 h, increase milk protein yield and erythrocyte glutathione, and reduce plasma urea nitrogen compared with Control. Lower plasma NEFA concentration and HSP70 mRNA expression in peripheral blood lymphocytes (P < 0.05) and tendencies towards greater plasma glucose concentration (P = 0.11) but less BW loss (P = 0.14) were observed in GY relative to CY cows. In conclusion, either CY or GY supplementation partially mitigated the negative effects of heat stress on performance and HSP70 mRNA expression of lactating cows, and GY supplementation provided additional improvements

  20. Thermostability of lactate dehydrogenase LDH-A4 isoenzyme: effect of heat shock protein DnaK on the enzyme activity.

    PubMed

    Zietara, M S; Skorkowski, E F

    1995-11-01

    Cells exposed to temperature a few degrees higher than their growth temperature synthesize heat shock proteins (hsp) which may then compose even 20% of total protein content. This paper examined the in vitro protective effect of heat shock protein DnaK (70 kDa) from Escherichia coli against the heat inactivation of lactate dehydrogenase isoenzyme LDH-A4. The LDH-A4 isoenzyme was purified from fish skeletal muscle using the affinity chromatography on Oxamate-agarose. The enzyme was then heated in the absence and the presence of DnaK protein in a water bath at either 51 or 55 degrees C. The LDH activity was determined by measuring the change in absorbency at 340 nm min-1 at 30 degrees C. The addition of DnaK protein to the LDH-A4 isoenzyme before heat treatment can protect enzyme activity against mild thermal inactivation. Incubation of the LDH-A4 isoenzyme at 51 degrees C in the presence of DnaK protein stimulates its activity by about 30%. The presence of 2 mM ATP can raise LDH activity by another 10%. No significant recovery was observed when DnaK protein was added to LDH at 25 degrees C following earlier inactivation. The maximal activities (Vmax) in the presence of DnaK protein are almost twice those without DnaK protein in the case of heat-treated LDH-A4 isoenzyme at 51 degrees C. The observed protection of LDH-A4 activity increased with the increasing DnaK protein concentration in the incubation medium. Results suggested that the presence of DnaK protein can protect LDH-A4 from heat inactivation.(ABSTRACT TRUNCATED AT 250 WORDS)

  1. Effects of feeding formate-treated alfalfa silage or red clover silage on omasal nutrient flow and microbial protein synthesis in lactating dairy cows.

    PubMed

    Brito, A F; Broderick, G A; Olmos Colmenero, J J; Reynal, S M

    2007-03-01

    Eight ruminally cannulated Holstein cows that were part of a larger lactation trial were blocked by days in milk and randomly assigned to replicated 4 x 4 Latin squares to quantify effects of nonprotein N (NPN) content of alfalfa silage (AS) and red clover silage (RCS) on omasal nutrient flows. Diets, fed as total mixed rations, contained 50% dry matter from control AS (CAS), ammonium tetraformate-treated AS (TAS), late maturity RCS (RCS1), or early maturity RCS (RCS2). Silages differed in NPN and acid detergent insoluble N (% of total N): 50 and 4% (CAS); 45 and 3% (TAS); 27 and 8% (RCS1); 29 and 4% (RCS2). The CAS, TAS, and RCS2 diets had 36% high-moisture shelled corn and 3% soybean meal, and the RCS1 diet had 31% high-moisture shelled corn and 9% soybean meal. All diets contained 10% corn silage, 27% neutral detergent fiber, and 17 to 18% crude protein. Compared with RCS, feeding AS increased the supply of rumen-degraded protein and omasal flows of nonammonia N and microbial protein, which may explain the improved milk yield observed in the companion lactation trial. However, omasal flow of rumen-undegraded protein was 34% greater on RCS. Except for Arg, omasal flows of individual AA, branched-chain AA, nonessential AA, essential AA, and total AA did not differ between cows fed AS vs. RCS. Within AS diets, no differences in omasal AA flows were observed. However, omasal flows of Asp, Ser, Glu, Cys, Val, Ile, Tyr, Lys, total nonessential AA, and total AA all were higher in cows fed RCS1 vs. cows fed RCS2. In this trial, there was no advantage to reducing NPN content of hay-crop silage.

  2. Ruminally protected lysine and methionine for lactating dairy cows fed a diet designed to meet requirements for microbial and postruminal protein.

    PubMed

    Robinson, P H; Fredeen, A H; Chalupa, W; Julien, W E; Sato, H; Fujieda, T; Suzuki, H

    1995-03-01

    Dairy cows, 20 at each of two sites, were used to determine responses to ruminally protected Lys and Met in a full lactation study. Cows were fed corn silage twice daily for ad libitum intake and a concentrate four times daily in proportion to milk production. At Truro, cows were fed 2.7 kg/d of alfalfa and timothy hay DM at 0600 and at 1500 h. At Fredericton, cows were fed 2.7 kg of timothy silage DM at 0600 h and 2.7 kg of alfalfa hay DM at 1500 h. Diets were designed to meet, but not to exceed, recommendations for ruminally degradable CP and intestinally digestible protein. Ten cows at each site were fed ruminally protected L-Lys.HCl (19 g/d) and DL-Met (6.5 g/d). Cows fed AA at each site produced more milk, lactose, protein, and fat; milk protein and fat percentages were also higher. No time x treatment interactions occurred for any production parameter. In spite of similar production responses between sites, cows fed AA consumed more DM at Truro, but those at Fredericton did not. Thus, gross efficiency of utilization of dietary N for milk N was increased with AA at Fredericton but not at Truro. However, considering the increased intake of CP by cows fed AA at Truro, an event that would have been expected to depress efficiency of utilization of dietary N, the lack of difference at Truro between treatments can be interpreted as an improvement, relative to expectations, because of AA feeding. High producing dairy cows fed a diet that was adequate in CP responded to ruminally protected Lys and Met primarily with increased production of milk protein and fat throughout the full lactation.

  3. Maternal protein restriction during pregnancy and lactation alters central leptin signalling, increases food intake, and decreases bone mass in 1 year old rat offspring.

    PubMed

    Qasem, Rani J; Li, Jing; Tang, Hee Man; Pontiggia, Laura; D'mello, Anil P

    2016-04-01

    The effects of perinatal nutrition on offspring physiology have mostly been examined in young adult animals. Aging constitutes a risk factor for the progressive loss of metabolic flexibility and development of disease. Few studies have examined whether the phenotype programmed by perinatal nutrition persists in aging offspring. Persistence of detrimental phenotypes and their accumulative metabolic effects are important for disease causality. This study determined the effects of maternal protein restriction during pregnancy and lactation on food consumption, central leptin sensitivity, bone health, and susceptibility to high fat diet-induced adiposity in 1-year-old male offspring. Sprague-Dawley rats received either a control or a protein restricted diet throughout pregnancy and lactation and pups were weaned onto laboratory chow. One-year-old low protein (LP) offspring exhibited hyperphagia. The inability of an intraperitoneal (i.p.) leptin injection to reduce food intake indicated that the hyperphagia was mediated by decreased central leptin sensitivity. Hyperphagia was accompanied by lower body weight suggesting increased energy expenditure in LP offspring. Bone density and bone mineral content that are negatively regulated by leptin acting via the sympathetic nervous system (SNS), were decreased in LP offspring. LP offspring did not exhibit increased susceptibility to high fat diet induced metabolic effects or adiposity. The results presented here indicate that the programming effects of perinatal protein restriction are mediated by specific decreases in central leptin signalling to pathways involved in the regulation of food intake along with possible enhancement of different CNS leptin signalling pathways acting via the SNS to regulate bone mass and energy expenditure.

  4. Melinjo (Gnetum gnemon) extract intake during lactation stimulates hepatic AMP-activated protein kinase in offspring of excessive fructose-fed pregnant rats.

    PubMed

    Kataoka, Saori; Mukai, Yuuka; Takebayashi, Mihoko; Kudo, Megumi; Acuram, Uson Rachael; Kurasaki, Masaaki; Sato, Shin

    2016-06-01

    Excessive maternal fructose intake during pregnancy and in early postnatal life has metabolic consequences for the offspring. We investigated the effects of melinjo (Gnetum gnemon) extract (MeE) intake during lactation on the expression and phosphorylation of adenosine monophosphate-activated protein kinase (AMPK) in the liver of offspring from excessive fructose-fed pregnant dams. Pregnant Wistar rats received a normal diet and 100g/L fructose solution during gestation ad libitum. At delivery, dams were divided into two groups: a control diet (FC) or a 0.1% MeE-containing diet (FM) fed during lactation. The dams that were not treated with fructose were fed a control diet (CC). At postnatal week 3, some pups were sacrificed, while the remaining continued to receive a normal diet and were sacrificed at week 17. Blood chemistry and phosphorylation levels of AMPK and acetyl-coenzyme A carboxylase (ACC) were evaluated. Plasma glucose levels in FC female offspring increased compared to that receiving CC at weeks 3 and 17; however, the levels in FM female offspring decreased at week 17. The insulin levels in FM female offspring decreased significantly compared to that in FC female offspring at week 3. Hepatic AMPK phosphorylation was upregulated in FM offspring at week 3 and in female, but not male, offspring at week 17. ACC phosphorylation in FM female offspring was upregulated at week 17. Our results suggest that maternal MeE intake during lactation may modulate the hepatic AMPK pathways in female offspring. Copyright © 2016 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

  5. Metabolic and productive response to ruminal protein degradability in early lactation cows fed untreated or xylose-treated soybean meal-based diets.

    PubMed

    Jahani-Moghadam, M; Amanlou, H; Nikkhah, A

    2009-12-01

    Effects of different dietary rumen undegradable (RUP) to degradable (RDP) protein ratios on ruminal nutrient degradation, feed intake, blood metabolites and milk production were determined in early lactation cows. Four multiparous (43 ± 5 days in milk) and four primiparous (40 ± 6 days in milk) tie-stall-housed Holstein cows were used in a duplicated 4 × 4 Latin square design with four 21-day periods. Each period had 14-day of adaptation and 7-day of sampling. Diets contained on a dry matter (DM) basis, 23.3% alfalfa hay, 20% corn silage and 56.7% concentrate. Cows were first offered alfalfa hay at 7:00, 15:00 and 23:00 hours, and 30 min after each alfalfa hay delivery were offered a mixture of corn silage and concentrate. Treatments were diets with RUP:RDP ratios of (i) 5.2:11.6 (control), (ii) 6.1:10.6, (iii) 7.1:9.5 and (iv) 8.1:8.5, on a dietary DM% basis. Different RUP:RDP ratios were obtained by partial and total replacement of untreated soybean meal (SBM) with xylose-treated SBM (XSBM). In situ study using three rumen-cannulated non-lactating cows showed that DM and crude protein (CP) of SBM had greater rapidly degradable fractions. The potentially degradable fractions were degraded more slowly in XSBM. Treatment cows produced greater milk, protein, lactose, solids-non-fat and total solids than control cows. Increasing RUP:RDP reduced blood urea linearly. Feed costs dropped at RUP:RDP ratios of 6.1:10.6 and 7.1:9.5, but not at 8.1:8.5, compared with the 5.2:11.6 ratio. Intake of DM and CP, rumen pH, blood glucose, albumin and total protein, faecal and urine pH, changes in body weight and body condition score, and milk lactose and solids-non-fat percentages did not differ among treatments. Results provide evidence that increasing dietary RUP:RDP ratio from 5.2:11.6 to 7.1:9.5 optimizes nitrogen metabolism and milk production and reduces feed costs in early lactation cows. Reduced blood urea suggests reprodutive benefits.

  6. Parathyroid hormone-related protein in lower vertebrates.

    PubMed

    Ingleton, P M

    2002-05-01

    The genes for parathyroid hormone-related protein (PTHrP) have been cloned in two teleost fishes, cDNA of sea bream (Sparus aurata) and genomic DNA of puffer fish (Fugu rubripes). The gene sequences show that there is significant conservation of amino acid identity, with specific domains most highly conserved. The N-terminus, responsible for bone matrix lysis in mammals and chickens, is present in the fish genes with 52% sequence identity to higher vertebrate PTHrP peptides; the nuclear transporter region shares 73% identity, and the RNA-binding sequence is 65% identical. However, the peptides are shorter then mammalian PTHrP, lacking the C-terminus responsible for inhibition of osteoclast lytic activity, but they have an additional inserted sequence between amino acids 38 and 54 that is not present in higher vertebrate PTHrPs. The N-terminus 1-38 Fugu PTHrP proved to be hypercalcaemic in larval Sparus, suggesting that it may be a physiological regulator of calcium homeostasis in fish. Using homologous nucleotide probes for in situ hybridisation and reverse-transcription polymerase chain reaction (RT-PCR) of extracted RNA, PTHrP gene expression has been widely found in both developing and adult fish. Antiserum to the fish insert sequence demonstrated transcription of PTHrP in all stages of Sparus development, and also detected the same epitope in tissues of developing frog (Rana temporaria), indicating that this has been retained during evolution of the amphibia.

  7. Participation of phosphatidyl inositol 3-kinase/protein kinase B and ERK1/2 pathways in interleukin-1beta stimulation of lactate production in Sertoli cells.

    PubMed

    Riera, María Fernanda; Galardo, María Noel; Pellizzari, Eliana Herminia; Meroni, Silvina Beatriz; Cigorraga, Selva Beatriz

    2007-04-01

    Interleukin-1beta (IL1beta ) belongs to a set of intratesticular regulators that provide the fine-tuning of cellular processes implicated in the maintenance of spermatogenesis. The aim of the present study was to analyze the signaling pathways that may participate in IL1beta regulation of Sertoli cell function. Sertoli cell cultures from 20-day-old rat were used. Stimulation of the cultures with IL1beta showed increments in phosphorylated protein kinase B (PKB), P70S6K, and ERK1/2 levels. A phosphatidyl inositol 3-kinase (PI3K) inhibitor (wortmannin (W)), a mammalian target of rapamycin inhibitor (rapamycin (R)), and a MEK inhibitor (PD98059 (PD)) were utilized to evaluate the participation of PI3K/PKB, P70S6K, and ERK1/2 pathways in the regulation of lactate production by IL1beta . PD and W, but not R, decreased IL1beta-stimulated lactate production. The participation of these pathways in the regulation of glucose uptake and lactate dehydrogenase (LDH) A mRNA levels by IL1beta was also analyzed. It was observed that W decreased IL1beta-stimulated glucose uptake, whereas PD and R did not modify it. On the other hand, PD decreased the stimulation of LDH A mRNA levels by IL1beta , whereas W and R did not modify it. In summary, results presented herein demonstrate that IL1beta stimulates PI3K/PKB-, P70S6K-, and ERK1/2-dependent pathways in rat Sertoli cells. Moreover, these results show that while IL1beta utilizes the PI3K/PKB pathway to regulate glucose transport, it utilizes the ERK1/2 pathway to regulate LDH A mRNA levels. This study reveals that IL1beta utilizes different signal transduction pathways to modify the biochemical steps that are important to regulate lactate production in rat Sertoli cells.

  8. Low-protein diet fed to crossbred sows during pregnancy and lactation enhances myostatin gene expression through epigenetic regulation in skeletal muscle of weaning piglets.

    PubMed

    Jia, Yimin; Gao, Guichao; Song, Haogang; Cai, Demin; Yang, Xiaojing; Zhao, Ruqian

    2016-04-01

    This study was aimed to investigate the effects of a maternal low-protein diet on transcriptional regulation of the myostatin (MSTN) gene in skeletal muscle of weaning piglets. Sows were fed either a standard-protein (SP, 15 and 18 % crude protein) or a low-protein (LP, 50 % protein level of SP) diet throughout pregnancy and lactation. Longissimus dorsi muscle was sampled from male piglets at 28 days of age. The mRNA was determined by RT-PCR, and protein was measured by Western blot. Chromatin immunoprecipitation assay was used to determine the binding of transcription factors and histone H3 modifications on the MSTN gene promoter. The maternal LP diet significantly decreased body weight and average daily gain (P < 0.05), which was associated with significantly lower plasma concentration of urea nitrogen and total protein (P < 0.05), as well as decreased muscle RNA content (P < 0.05). MSTN mRNA (P < 0.05) was significantly increased, together with enhanced (P < 0.05) mRNA and protein expression of forkhead box class O family member protein 3 (FoxO3), and a tendency of an increase (P = 0.10) in glucocorticoid receptor (GR) mRNA in the muscle of LP piglets. Furthermore, the binding of both FoxO3 and GR to the MSTN gene promoter was significantly higher (P < 0.05) in muscle of LP piglets, together with significantly enriched (P < 0.05) gene activation markers, H3K9Ac and H3K4me3. These results indicate that MSTN mediates maternal LP diet-induced growth retardation, through epigenetic regulation involving FoxO3 and GR binding to its promoter.

  9. Urea-N recycling in lactating dairy cows fed diets with 2 different levels of dietary crude protein and starch with or without monensin.

    PubMed

    Recktenwald, E B; Ross, D A; Fessenden, S W; Wall, C J; Van Amburgh, M E

    2014-03-01

    Rumensin (monensin; Elanco Animal Health, Greenfield, IN) has been shown to reduce ammonia production and microbial populations in vitro; thus, it would be assumed to reduce ruminal ammonia production and subsequent urea production and consequently affect urea recycling. The objective of this experiment was to determine the effects of 2 levels of dietary crude protein (CP) and 2 levels of starch, with and without Rumensin on urea-N recycling in lactating dairy cattle. Twelve lactating Holstein dairy cows (107 ± 21 d in milk, 647 kg ± 37 kg of body weight) were fed diets characterized as having high (16.7%) or low (15.3%) CP with or without Rumensin, while dietary starch levels (23 vs. 29%) were varied between 2 feeding periods with at least 7d of adaptation between measurements. Cows assigned to high or low protein and to Rumensin or no Rumensin remained on those treatments to avoid carryover effects. The diets consisted of approximately 40% corn silage, 20% alfalfa hay, and 40% concentrate mix specific to the treatment diets, with 0.5 kg of wheat straw added to the high starch diets to enhance effective fiber intake. The diets were formulated using Cornell Net Carbohydrate and Protein System (version 6.1), and the low-protein diets were formulated to be deficient for rumen ammonia to create conditions that should enhance the demand for urea recycling. The high-protein diets were formulated to be positive for both rumen ammonia and metabolizable protein. Rumen fluid, urine, feces, and milk samples were collected before and after a 72-h continuous jugular infusion of (15)N(15)N-urea. Total urine and feces were collected during the urea infusions for N balance measurements. Milk yield and dry matter intake were improved in cows fed the higher level of dietary CP and by Rumensin. Ruminal ammonia and milk and plasma urea nitrogen concentrations corresponded to dietary CP concentration. As has been shown in vitro, Rumensin reduced rumen ammonia concentration by

  10. Lactate Dehydrogenase 5 Expression in Non-Hodgkin Lymphoma Is Associated with the Induced Hypoxia Regulated Protein and Poor Prognosis

    PubMed Central

    Chen, Zhujun; Xu, Xiaofeng; Hu, Hongfeng; Zhao, Xinmin; Gao, Xiang; Guo, Lin

    2013-01-01

    Lactate dehydrogenase 5 (LDH-5) is one of the major isoenzymes catalyzing the biochemical process of pyruvate to lactate. The purpose of this study was to investigate the expression of serum LDH-5 and test whether this enzyme is regulated by tumor hypoxia and represents a prognostic marker in patients with Non-Hodgkin’s lymphoma (NHL). In this study, LDH-5 levels were detected using agarose gel electrophoresis in NHL patients (n = 266) and non-NHL controls including benign lymphadenectasis (n = 30) and healthy cohorts (n = 233). We also explored the expression of LDH-5 and hypoxia-inducible factor (HIF) 1α in NHL and benign controls by immunohistochemistry and immunofluorescence staining, respectively. Moreover, the role of LDH-5 in the progression of NHL was assessed by multivariate Cox analyses and Kaplan-Meier survival estimates. Serum concentrations of LDH-5 were significantly higher in NHL patients (9.3%) than in benign patients and healthy controls (7.5% and 7.2%, respectively, P<0.01). Application of LDH-5 detection increased the sensitivity of NHL detection, identifying 53.4% of NHL patients as positive, compared with the measurement of total LDH levels (36.5% sensitivity). LDH-5 concentrations increased with clinical stage, extra-nodal site involvement, and WHO performance status of patients with NHL. Exposure to a hypoxic environment induced the expression of LDH-5 and its overexpression correlated with HIF1α cytoplasmic accumulation in NHL cells. In multivariate analyses, LDH-5 was an independent marker for progression-free survival in patients with NHL (P<0.001). Overall, the expression of LDH-5 was elevated in NHL, showing an association with tumor hypoxia and unfavorable prognosis. Thus, LDH-5 emerges as a promising prognostic predictor for NHL patients. PMID:24086384

  11. Parathyroid hormone-related protein inhibits DKK1 expression through c-Jun-mediated inhibition of β-catenin activation of the DKK1 promoter in prostate cancer.

    PubMed

    Zhang, H; Yu, C; Dai, J; Keller, J M; Hua, A; Sottnik, J L; Shelley, G; Hall, C L; Park, S I; Yao, Z; Zhang, J; McCauley, L K; Keller, E T

    2014-05-08

    Prostate cancer (PCa)bone metastases are unique in that majority of them induce excessive mineralized bone matrix, through undefined mechanisms, as opposed to most other cancers that induce bone resorption. Parathyroid hormone-related protein (PTHrP) is produced by PCa cells and intermittent PTHrP exposure has bone anabolic effects, suggesting that PTHrP could contribute to the excess bone mineralization. Wnts are bone-productive factors produced by PCa cells, and the Wnt inhibitor Dickkopfs-1 (DKK1) has been shown to promote PCa progression. These findings, in conjunction with the observation that PTHrP expression increases and DKK1 expression decreases as PCa progresses, led to the hypothesis that PTHrP could be a negative regulator of DKK1 expression in PCa cells and, hence, allow the osteoblastic activity of Wnts to be realized. To test this, we first demonstrated that PTHrP downregulated DKK1 mRNA and protein expression. We then found through multiple mutated DKK1 promoter assays that PTHrP, through c-Jun activation, downregulated the DKK1 promoter through a transcription factor (TCF) response element site. Furthermore, chromatin immunoprecipitation (ChIP) and re-ChIP assays revealed that PTHrP mediated this effect through inducing c-Jun to bind to a transcriptional activator complex consisting of β-catenin, which binds the most proximal DKK1 promoter, the TCF response element. Together, these results demonstrate a novel signaling linkage between PTHrP and Wnt signaling pathways that results in downregulation of a Wnt inhibitor allowing for Wnt activity that could contribute the osteoblastic nature of PCa.

  12. Effects of Protein Level and Mangosteen Peel Pellets (Mago-pel) in Concentrate Diets on Rumen Fermentation and Milk Production in Lactating Dairy Crossbreds

    PubMed Central

    Norrapoke, T.; Wanapat, M.; Wanapat, S.

    2012-01-01

    Four, lactating dairy crossbreds (50%×50% Holstein Friesian×Native Zebu cattle) were randomly assigned according to a 2×2 factorial arrangement (two protein levels and two levels of mangosteen peel pellets (Mago-pel)) in a 4×4 Latin square design to receive four dietary treatments. All cows received concentrate at a proportion of 1 kg concentrate per 2 kg of milk yield, and urea-treated 5% rice straw (UTRS) was given ad libitum. It was found that total dry matter intakes, nutrient digestibility, ruminal pH and NH3-N concentrations were not affected (p>0.05) by treatments. Concentrations of ruminal pH and NH3-N were not affected by dietary treatments although the concentration of BUN varied significantly (p<0.05) between protein levels (p<0.05). The populations of rumen bacteria and fungal zoospores did not differ among treatments (p>0.05); however, the population of protozoa was decreased (p<0.05) when cows received Mago-pel supplementation. The composition of the population of bacteria, identified by real-time PCR technique, including total bacteria, methanogens, Fibrobacter succinogenes and Ruminococcus albus was similar (p>0.05) among dietary treatments (p>0.05); however, copy numbers of Ruminococcus flavefaciens was increased when protein level increased (p<0.05). Microbial protein synthesis, in terms of both quantity and efficiency, was enriched by Mago-pel supplementation. Milk yield was greatest in cows fed UTRS based diets with concentrate containing protein at 16% CP with Mago-pel, but were lowest without Mago-pel (p<0.05). In addition, protein level and supplementation of Mago-pel did not affect (p>0.05) milk composition except solids-not-fat which was higher in cows fed the diet with 19% CP. Therefore, feeding a concentrate containing 16% CP together with 300 g/hd/d Mago-pel supplementation results in changes in rumen fermentation and microbial population and improvements in milk production in lactating dairy crossbreds fed on UTRS. PMID:25049652

  13. Effect of supplemental concentrate during the dry period or early lactation on rumen epithelium gene and protein expression in dairy cattle during the transition period.

    PubMed

    Dieho, K; van Baal, J; Kruijt, L; Bannink, A; Schonewille, J T; Carreño, D; Hendriks, W H; Dijkstra, J

    2017-09-01

    We previously reported 2 experiments with rumen-cannulated Holstein-Friesian dairy cows showing that during the transition period, rumen papillae surface area, and fractional absorption rate of volatile fatty acids (VFA) increase after calving. However, supplemental concentrate during the dry period and rate of increase of concentrate allowance during lactation affected papillae surface area, but not VFA absorption. Here we report the changes in gene and protein expression in rumen papillae related to tissue growth and VFA utilization. The lactation experiment treatment consisted of a rapid [RAP; 1.0 kg of dry matter (DM)/d; n = 6] or gradual (GRAD; 0.25 kg of DM/d; n = 6) increase of concentrate allowance (up to 10.9 kg of DM/d), starting at 4 d postpartum (pp). The dry period experiment treatment consisted of 3.0 kg of DM/d of concentrate (n = 4) or no concentrate (n = 5) during the last 28 d of the dry period. Real-time quantitative PCR analysis of rumen papillae showed that the expression of apoptosis-related genes was neither affected by day nor its interaction with treatment for both experiments. Expression of epithelial transporter genes was not affected by day or treatment in the lactation experiment, except for NBC1. In the dry period experiment, expression of MCT1, NBC1, DRA, NHE2, NHE3, and UT-B generally decreased after calving. A day and treatment interaction was observed for ATP1A1 in the dry period experiment, with greater expression at 18 and 8 d antepartum for concentrate than no concentrate. Generally, expression of VFA metabolism-related genes was not affected by day or its interaction with treatment. In the lactation experiment, immunoblotting of 5 selected genes showed that protein expression of DRA and PCCA was greater at 16 d pp compared with 3 and 44 d pp. Expression of NHE2 was greater, and that of ATP1A1 lower, at 16 and 44 d pp compared with 3 d pp, suggesting alterations in intracellular pH regulation and sodium homeostasis. Both MCT1

  14. Molecular characterisation and expression of parathyroid hormone-related protein in the caudal neurosecretory system of the euryhaline flounder, Platichthys flesus.

    PubMed

    Lu, Weiqun; Jin, Yingying; Xu, Jinling; Greenwood, Michael P; Balment, Richard J

    2017-02-27

    Parathyroid hormone-related protein (PTHrP) is a hypercalcemic factor in fish, but the source of circulating PTHrP remains unclear. In this study investigation of the caudal neurosecretory system (CNSS), considered one of major sources of PTHrP in fish, provided valuable insights into this regulatory system. We report pthrpa and pthrpb gene cloning, characterization, expression, and responses to low salinity and hypocalcemia challenge in flounder. The pthrpa and pthrpb precursors, isolated from a European flounder CNSS library, consist of 166 and 192 amino acid residues, respectively, with an overall homology of approximately 59.2%. Both precursors contain a signal peptide and a mature peptide with cleavage and amidation sites. The flounder PTHrPA and PTHrPB peptides share only 41% sequence identity with human PTHrPA. Quantitative PCR analysis demonstrated that the bone and bladder, are respectively major sites of pthrpa and pthrpb expression in flounder. Urophysectomy confirmed the CNSS as a likely contributor to circulating PTHrP peptides. There were no significant differences in CNSS pthrpa and pthrpb mRNA expression or plasma PTHrP levels between seawater (SW) and freshwater (FW)-adapted fish, though plasma total calcium concentrations were higher in FW animals. The intraperitonial administration of EGTA rapidly induced hypocalcemia and concomitant elevation in plasma PTHrP accompanied by increases in both pthrpa and pthrpb expression in the CNSS. Together, these findings support an evolutionary conserved role for PTHrP in the endocrine regulation of calcium.

  15. Parathyroid Hormone-Related Protein, Human Adipose-Derived Stem Cells Adipogenic Capacity and Healthy Obesity.

    PubMed

    Roca-Rodríguez, María Mar; El Bekay, Rajaa; Garrido-Sanchez, Lourdes; Gómez-Serrano, María; Coin-Aragüez, Leticia; Oliva-Olivera, Wilfredo; Lhamyani, Said; Clemente-Postigo, Mercedes; García-Santos, Eva; de Luna Diaz, Resi; Yubero-Serrano, Elena M; Fernández Real, José M; Peral, Belén; Tinahones, Francisco J

    2015-06-01

    This study aimed to define the potential role of PTHrP on adipogenic regulation and to analyze its relationship with obesity and insulin resistance. This was a cross-sectional study in which visceral (VAT) and subcutaneous (SAT) adipose tissue were extracted from 19 morbidly obese, 10 obese, and 10 lean subjects. PTHrP mRNA levels were measured in VAT and SAT. VAT mesenchymal stem cells and 3T3-L1 cells were differentiated into adipocytes in presence or absence of PTHrP siRNA. PTHrP mRNA and protein levels as well as adipogenic markers were evaluated by Western blotting or qPCR. Immunohistochemistry and immunofluorescence procedures were used for PTHrP intracellular localization. Both human VAT and SAT express PTHrP protein mainly in the nucleolar compartment of stromal vascular fraction cells. The highest levels of PTHrP mRNA and protein expression were detected in undifferentiated mesenchymal cells and progressively decreased during adipogenesis. Remarkably, adipogenic differentiation in human mesenchymal stem cells (A-hMSC) was significantly impaired in a pthrp knockdown. PTHrP seems to be related to obesity-associated insulin resistance (IR), given that we found that PTHrP mRNA expression was higher in VAT from morbidly obese with a low IR degree (MO-L-IR) subjects than those from morbidly obese with a high IR degree (MO-H-IR) and lean subjects, and correlated positively with body mass index and hip circumference. We also found that A-hMSC from MO-L-IRs displayed higher adipogenic capacity than those from both MO-H-IRs and leans. In addition, adipogenesis was impaired in VAT from MO-H-IRs, given that mRNA expression levels of key adipogenic regulators were lower than those from MO-L-IR subjects. PTHrP could be a potential new therapeutic target for the reprograming of adipogenesis and adipose tissue expansion, thus possibly ameliorating the metabolic syndrome in obese subjects.

  16. Effects of corn-based diet starch content and neutral detergent fiber source on lactation performance, digestibility, and bacterial protein flow in dairy cows.

    PubMed

    Fredin, S M; Akins, M S; Ferraretto, L F; Shaver, R D

    2015-01-01

    An experiment was conducted to evaluate the effects of corn-based dietary starch content and source of neutral detergent fiber (NDF) on lactation performance, nutrient digestion, bacterial protein flow, and ruminal parameters in lactating dairy cows. Eight ruminally cannulated multiparous Holstein cows averaging 193±11d in milk were randomly assigned to treatments in a replicated 4×4 Latin square design with 21-d periods. Treatment diets were high corn grain (HCG; 38% corn silage, 19% dry ground corn, and 4% soy hulls), high soy hulls (HSH; 38% corn silage, 11% dry ground corn, and 13% soy hulls), high corn silage (HCS; 50% corn silage, 6% dry ground corn, and 4% soy hulls), and low corn silage (LCS; 29% corn silage, 15% corn, and 19% soy hulls). The HCG, HSH, HCS, and LCS diets contained 29, 23, 24, and 22% starch; 27, 32, 30, and 32% total NDF; and 21, 21, 25, and 17% forage NDF (dry matter basis), respectively. Mean dry matter intake and milk yield were unaffected by treatment. Cows fed LCS had reduced milk fat content compared with HSH and HCS. The concentration of milk urea nitrogen was greater for cows fed HCS compared with the other treatments. Total-tract digestion of NDF was reduced for cows fed the HCG diet. Total-tract starch digestion was increased for cows fed the HSH and HCS compared with HCG and LCS diets. Bacterial protein flow was unaffected by treatment. Ruminal ammonia concentration was reduced in cows fed the HCG and LCS diets compared with the HCS diet. Ruminal propionate increased and the acetate:propionate ratio decreased in cows fed the LCS diet compared with the HCS diet. Ruminal pH was greater for cows fed the HCS diet compared with cows fed the LCS diet. Diet digestibility and performance of mid- to late-lactation cows fed reduced-starch diets by partially replacing corn grain with soy hulls or corn silage was similar to or improved compared with cows fed a normal-starch diet.

  17. Hindbrain lactate regulates preoptic gonadotropin-releasing hormone (GnRH) neuron GnRH-I protein but not AMPK responses to hypoglycemia in the steroid-primed ovariectomized female rat.

    PubMed

    Shrestha, P K; Briski, K P

    2015-07-09

    Steroid positive-feedback activation of the gonadotropin-releasing hormone (GnRH)-pituitary luteinizing hormone (LH) neuroendocrine axis propagates the pre ovulatory LH surge, a crucial component of female reproduction. Our work shows that this key event is restrained by inhibitory metabolic input from hindbrain A2 noradrenergic neurons. GnRH neurons express the ultra-sensitive energy sensor adenosine 5'-monophosphate-activated protein kinase (AMPK); here, we investigated the hypothesis that GnRH nerve cell AMPK and peptide neurotransmitter responses to insulin-induced hypoglycemia are controlled by hindbrain lack of the oxidizable glycolytic end-product L-lactate. Data show that hypoglycemic inhibition of LH release in steroid-primed ovariectomized female rats was reversed by coincident caudal hindbrain lactate infusion. Western blot analyses of laser-microdissected A2 neurons demonstrate hypoglycemic augmentation [Fos, estrogen receptor-beta (ER-β), phosphoAMPK (pAMPK)] and inhibition (dopamine-beta-hydroxylase, GLUT3, MCT2) of protein expression in these cells, responses that were normalized by insulin plus lactate treatment. Hypoglycemia diminished rostral preoptic GnRH nerve cell GnRH-I protein and pAMPK content; the former, but not the latter response was reversed by lactate. Results implicate caudal hindbrain lactoprivic signaling in hypoglycemia-induced suppression of the LH surge, demonstrating that lactate repletion of that site reverses decrements in A2 catecholamine biosynthetic enzyme and GnRH neuropeptide precursor protein expression. Lack of effect of lactate on hypoglycemic patterns of GnRH AMPK activity suggests that this sensor is uninvolved in metabolic-inhibition of positive-feedback-stimulated hypophysiotropic signaling to pituitary gonadotropes.

  18. Insulin, CCAAT/Enhancer-Binding Proteins and Lactate Regulate the Human 11β-Hydroxysteroid Dehydrogenase Type 2 Gene Expression in Colon Cancer Cell Lines

    PubMed Central

    Alikhani-Koupaei, Rasoul; Ignatova, Irena D.; Guettinger, Andreas; Frey, Felix J.; Frey, Brigitte M.

    2014-01-01

    11β-Hydroxysteroid dehydrogenases (11beta-HSD) modulate mineralocorticoid receptor transactivation by glucocorticoids and regulate access to the glucocorticoid receptor. The isozyme 11beta-HSD2 is selectively expressed in mineralocorticoid target tissues and its activity is reduced in various disease states with abnormal sodium retention and hypertension, including the apparent mineralocorticoid excess. As 50% of patients with essential hypertension are insulin resistant and hyperinsulinemic, we hypothesized that insulin downregulates the 11beta-HSD2 activity. In the present study we show that insulin reduced the 11beta-HSD2 activity in cancer colon cell lines (HCT116, SW620 and HT-29) at the transcriptional level, in a time and dose dependent manner. The downregulation was reversible and required new protein synthesis. Pathway analysis using mRNA profiling revealed that insulin treatment modified the expression of the transcription factor family C/EBPs (CCAAT/enhancer-binding proteins) but also of glycolysis related enzymes. Western blot and real time PCR confirmed an upregulation of C/EBP beta isoforms (LAP and LIP) with a more pronounced increase in the inhibitory isoform LIP. EMSA and reporter gene assays demonstrated the role of C/EBP beta isoforms in HSD11B2 gene expression regulation. In addition, secretion of lactate, a byproduct of glycolysis, was shown to mediate insulin-dependent HSD11B2 downregulation. In summary, we demonstrate that insulin downregulates HSD11B2 through increased LIP expression and augmented lactate secretion. Such mechanisms are of interest and potential significance for sodium reabsorption in the colon. PMID:25133511

  19. The effect of high and low levels of supplementation on milk production, nitrogen utilization efficiency, and milk protein fractions in late-lactation dairy cows.

    PubMed

    Reid, M; O'Donovan, M; Murphy, J P; Fleming, C; Kennedy, E; Lewis, E

    2015-08-01

    To fill the feed deficit in the autumn/late lactation period in a seasonal grazing system, supplementation is required. This study aimed to investigate the use of baled grass silage or concentrate as supplementation to grazing dairy cows in late lactation. Eighty-four grass-based spring-calving dairy cows, averaging 212d in milk, were allocated to 1 of 6 treatments [high grass allowance (HG), low grass allowance (LG), grass with a low concentrate allocation (GCL), grass with a low grass silage allocation (GSL), grass with a high concentrate allocation (GCH), and grass with a high grass silage allocation (GSH)] to measure the effects of using baled grass silage or concentrate as supplements to grazed grass. Effects on intake, milk yield, milk composition and N fractions, and N utilization efficiency were measured. Treatments HG and LG received 17 and 14kg of dry matter (DM) grass/cow per d, respectively. Treatments GCL and GSL were offered 14kg of DM grass/cow per d and 3kg of DM of supplementation/cow per d. Treatments GCH and GSH were offered 11kg of DM grass/cow per d and 6kg of DM of supplementation/cow per d. Milk yield was greatest in the GCH treatment and milk solids yield was greatest in both concentrate-supplemented treatments. The HG and LG treatments excreted a greater quantity of N as a proportion of N intake than the supplemented treatments. The HG treatment also excreted the greatest total quantity of N. This indicates an improvement in N utilization efficiency when supplementation is offered compared with grazing only. Offering 6kg of DM of either grass silage or concentrate as supplementation decreased milk true protein concentration compared with offering a grass-only diet. This suggests that increasing the proportion of supplementation relative to grass may negatively affect milk processability, which is associated with milk true protein concentration.

  20. Maternal high fructose and low protein consumption during pregnancy and lactation share some but not all effects on early-life growth and metabolic programming of rat offspring.

    PubMed

    Arentson-Lantz, Emily J; Zou, Mi; Teegarden, Dorothy; Buhman, Kimberly K; Donkin, Shawn S

    2016-09-01

    Maternal nutritional stress during pregnancy acts to program offspring metabolism. We hypothesized that the nutritional stress caused by maternal fructose or low protein intake during pregnancy would program the offspring to develop metabolic aberrations that would be exacerbated by a diet rich in fructose or fat during adult life. The objective of this study was to characterize and compare the fetal programming effects of maternal fructose with the established programming model of a low-protein diet on offspring. Male offspring from Sprague-Dawley dams fed a 60% starch control diet, a 60% fructose diet, or a low-protein diet throughout pregnancy and lactation were weaned onto either a 60% starch control diet, 60% fructose diet, or a 30% fat diet for 15 weeks. Offspring from low-protein and fructose-fed dam showed retarded growth (P<.05) at weaning (50.3, 29.6 vs 59.1±0.8 g) and at 18 weeks of age (420, 369 vs 464±10.9 g). At 18 weeks of age, offspring from fructose dams expressed greater quantities (P<.05) of intestinal Pgc1a messenger RNA compared with offspring from control or low-protein dams (1.31 vs 0.89, 0.85; confidence interval, 0.78-1.04). Similarly, maternal fructose (P=.09) and low-protein (P<.05) consumption increased expression of Pgc1a in offspring liver (7.24, 2.22 vs 1.22; confidence interval, 2.11-3.45). These data indicate that maternal fructose feeding is a programming model that shares some features of maternal protein restriction such as retarded growth, but is unique in programming of selected hepatic and intestinal transcripts. Copyright © 2016. Published by Elsevier Inc.

  1. Messenger RNA stability of parathyroid hormone-related protein regulated by transforming growth factor-beta1.

    PubMed

    Sellers, R S; Capen, C C; Rosol, Thomas J

    2002-02-25

    Humoral hypercalcemia of malignancy (HHM), a paraneoplastic syndrome associated with epithelial cancers, including squamous cell carcinoma (SCC), is due to expression and secretion of parathyroid hormone-related protein (PTHrP). Transforming growth factor-beta1 (TGFbeta1), expressed by many tumors, has been demonstrated in vitro to increase the half-life of PTHrP mRNA. In this study, oral squamous carcinoma cells (SCC2/88) had a two-fold increase in PTHrP mRNA stability (from 45 to 90 min) in response to treatment with TGFbeta1. In order to examine the mechanism of TGFbeta1-mediated PTHrP mRNA stability, a cell-free assay of mRNA degradation was utilized in which the degradation of in vitro-transcribed mRNA incubated with cytoplasmic protein extracts from SCC2/88 treated with vehicle or TGFbeta1 was measured. In this assay, full-length PTHrP mRNA was not significantly stabilized in TGFbeta1-treated samples when compared to vehicle treated samples. However, there was a striking (>5-fold) increase in PTHrP mRNA half-life in TGFbeta1-treated samples when PTHrP mRNA lacked the 3'-untranslated region (3'-UTR). In contrast, the degradation of 3'-UTR-truncated PTHrP mRNA using the cell-free assay was not altered in vehicle-treated samples. UV cross-linking of PTHrP mRNA and cytoplasmic proteins from cells treated with either vehicle or TGFbeta1 revealed numerous mRNA-binding proteins. TGFbeta1 treatment resulting in decreased binding of 33, 31, 27, 20 and 18 kDa binding proteins to the terminal coding region. These studies revealed that TGFbeta1-induced PTHrP mRNA stability might be, in part, the result of cis-acting sequences within the coding region of the PTHrP mRNA.

  2. Lactate metabolism in the fetal rabbit lung

    SciTech Connect

    Engle, M.J.; Brown, D.J.; Dooley, M.

    1986-05-01

    Lactate is frequently overlooked as a potential substrate for the fetal lung, even though it is present in the fetal circulation in concentrations as high as 8 mM. These high concentrations, coupled with the relatively low levels of glucose in the fetal blood, may indicate that lactate can substitute for glucose in pulmonary energy generation and phospholipid synthesis. A series of experiments was therefore undertaken in order to investigate the role of lactate in perinatal pulmonary development. Explants from 30 day gestation fetal rabbit lungs were incubated in Krebs-Ringer bicarbonate buffer supplemented with 3 mM (U-/sup 14/C)-glucose and varying levels of lactate. In the absence of medium lactate, fetal rabbit lung explants were capable of producing lactate at a rate of approximately 200 etamoles/mg protein/hour. The addition of lactate to the bathing medium immediately reduced net lactate production and above 4 mM, fetal rabbit lung explants became net utilizers of lactate. Media lactate concentrations of 2.5 mM, 5 mM and 10 mM also decreased glucose incorporation into total tissue disaturated phosphatidylcholine by approximately 20%, 35%, and 45%, respectively. Glucose incorporation into surfactant phosphatidylcholine was also reduced by approximately 50%, when lactate was present in the incubation medium at a concentration of 5 mM. Additional experiments also revealed that fetal lung lactate dehydrogenase activity was almost twice that found in the adult rabbit lung. These data indicate that lactate may be an important carbon source for the developing lung and could be a significant component in the manufacture of surfactant phosphatidylcholine during late gestation.

  3. The intrinsically disordered protein LEA7 from Arabidopsis thaliana protects the isolated enzyme lactate dehydrogenase and enzymes in a soluble leaf proteome during freezing and drying.

    PubMed

    Popova, Antoaneta V; Rausch, Saskia; Hundertmark, Michaela; Gibon, Yves; Hincha, Dirk K

    2015-10-01

    The accumulation of Late Embryogenesis Abundant (LEA) proteins in plants is associated with tolerance against stresses such as freezing and desiccation. Two main functions have been attributed to LEA proteins: membrane stabilization and enzyme protection. We have hypothesized previously that LEA7 from Arabidopsis thaliana may stabilize membranes because it interacts with liposomes in the dry state. Here we show that LEA7, contrary to this expectation, did not stabilize liposomes during drying and rehydration. Instead, it partially preserved the activity of the enzyme lactate dehydrogenase (LDH) during drying and freezing. Fourier-transform infrared (FTIR) spectroscopy showed no evidence of aggregation of LDH in the dry or rehydrated state under conditions that lead to complete loss of activity. To approximate the complex influence of intracellular conditions on the protective effects of a LEA protein in a convenient in-vitro assay, we measured the activity of two Arabidopsis enzymes (glucose-6-P dehydrogenase and ADP-glucose pyrophosphorylase) in total soluble leaf protein extract (Arabidopsis soluble proteome, ASP) after drying and rehydration or freezing and thawing. LEA7 partially preserved the activity of both enzymes under these conditions, suggesting its role as an enzyme protectant in vivo. Further FTIR analyses indicated the partial reversibility of protein aggregation in the dry ASP during rehydration. Similarly, aggregation in the dry ASP was strongly reduced by LEA7. In addition, mixtures of LEA7 with sucrose or verbascose reduced aggregation more than the single additives, presumably through the effects of the protein on the H-bonding network of the sugar glasses.

  4. Effects of corn-based diet starch content and corn particle size on lactation performance, digestibility, and bacterial protein flow in dairy cows.

    PubMed

    Fredin, S M; Ferraretto, L F; Akins, M S; Bertics, S J; Shaver, R D

    2015-01-01

    An experiment was conducted to determine the effects of dietary starch content in corn-based diets and corn particle size on lactation performance, nutrient digestibility, and bacterial protein flow in dairy cows using the omasal and reticular sampling technique. Eight ruminally cannulated lactating multiparous Holstein cows were used in a replicated 4×4 Latin square design with a 2×2 factorial arrangement of treatments. Treatments were fine (FG; mean particle size=552µm) and coarse (CG; 1,270µm) ground dry shelled corn in normal- (NS) and reduced- (RS) starch diets fed as total mixed rations. The NS and RS rations contained 27 and 18% starch (dry matter basis), respectively, and were formulated by partially replacing corn with soy hull pellets. Mean dry matter intake was unaffected by treatment (23.2kg/d). Cows fed NS diets produced 1.9kg/d more milk and 0.06kg/d more milk protein compared with cows fed RS diets. Cows fed NSFG and RSCG diets produced more fat-corrected milk than did cows fed NSCG and RSFG diets. Milk urea concentration was decreased for cows fed NS diets (12.4mg/dL) compared with RS diets (13.5mg/dL). Ruminal digestibility of neutral detergent fiber (NDF; % of NDF intake) determined by the omasal sampling technique was increased in cows fed RS diets compared with NS diets (43.4 vs. 34.9%), and total-tract digestibility of NDF (% of NDF intake) was increased in cows fed RS diets compared with those fed NS diets (50.1 vs. 43.1%). Ruminal digestibility of starch (% of starch intake) determined by the omasal sampling technique was greater in cows fed NS diets compared with those fed RS diets (85.6 vs. 81.6%). Total-tract starch digestion was increased in cows fed RS diets compared with those fed NS diets (96.9 vs. 94.6%) and in cows fed FG diets compared with those fed CG diets (98.0 vs. 93.5%). Bacterial protein flow was unaffected by treatment. The omasal and reticular sampling techniques resulted in similar treatment effects for nutrient flow

  5. FGF-21 and Skeletal Remodeling During and After Lactation in C57BL/6J Mice

    PubMed Central

    Bornstein, Sheila; Brown, Sue A.; Le, Phuong T.; Wang, Xunde; DeMambro, Victoria; Horowitz, Mark C.; MacDougald, Ormond; Baron, Roland; Lotinun, Sutada; Karsenty, Gerard; Wei, Wei; Ferron, Mathieu; Kovacs, Christopher S.; Clemmons, David

    2014-01-01

    Lactation is associated with significant alterations in both body composition and bone mass. Systemic and local skeletal factors such as receptor activator of nuclear factor κ-B ligand (RANKL), PTHrP, calcitonin, and estrogen are known to regulate bone remodeling during and after lactation. Fibroblast growth factor 21 (FGF-21) may function as an endocrine factor to regulate body composition changes during lactation by inducing gluconeogenesis and fatty acid oxidation. In this study, we hypothesized that the metabolic changes during lactation were due in part to increased circulating FGF-21, which in turn could accentuate bone loss. We longitudinally characterized body composition in C57BL/6J (B6) mice during (day 7 and day 21 of lactation) and after normal lactation (day 21 postlactation). At day 7 of lactation, areal bone density declined by 10% (P < .001), bone resorption increased (P < .0001), percent fat decreased by 20%, energy expenditure increased (P < .01), and markers of brown-like adipogenesis were suppressed in the inguinal depot and in preformed brown adipose tissue. At day 7 of lactation there was a 2.4-fold increase in serum FGF-21 vs baseline (P < .0001), a 8-fold increase in hepatic FGF-21 mRNA (P < .03), a 2-fold increase in undercarboxylated osteocalcin (Glu13 OCn) (P < .01), and enhanced insulin sensitivity. Recovery of total areal bone density was noted at day 21 of lactation, whereas the femoral trabecular bone volume fraction was still reduced (P < .01). Because FGF-21 levels rose rapidly at day 7 of lactation in B6 lactating mice, we next examined lactating mice with a deletion in the Fgf21 gene. Trabecular and cortical bone masses were maintained throughout lactation in FGF-21−/− mice, and pup growth was normal. Compared with lactating control mice, lactating FGF-21−/− mice exhibited an increase in bone formation, but no change in bone resorption. In conclusion, in addition to changes in calciotropic hormones, systemic FGF-21 plays

  6. FGF-21 and skeletal remodeling during and after lactation in C57BL/6J mice.

    PubMed

    Bornstein, Sheila; Brown, Sue A; Le, Phuong T; Wang, Xunde; DeMambro, Victoria; Horowitz, Mark C; MacDougald, Ormond; Baron, Roland; Lotinun, Sutada; Karsenty, Gerard; Wei, Wei; Ferron, Mathieu; Kovacs, Christopher S; Clemmons, David; Wan, Yihong; Rosen, Clifford J

    2014-09-01

    Lactation is associated with significant alterations in both body composition and bone mass. Systemic and local skeletal factors such as receptor activator of nuclear factor κ-B ligand (RANKL), PTHrP, calcitonin, and estrogen are known to regulate bone remodeling during and after lactation. Fibroblast growth factor 21 (FGF-21) may function as an endocrine factor to regulate body composition changes during lactation by inducing gluconeogenesis and fatty acid oxidation. In this study, we hypothesized that the metabolic changes during lactation were due in part to increased circulating FGF-21, which in turn could accentuate bone loss. We longitudinally characterized body composition in C57BL/6J (B6) mice during (day 7 and day 21 of lactation) and after normal lactation (day 21 postlactation). At day 7 of lactation, areal bone density declined by 10% (P < .001), bone resorption increased (P < .0001), percent fat decreased by 20%, energy expenditure increased (P < .01), and markers of brown-like adipogenesis were suppressed in the inguinal depot and in preformed brown adipose tissue. At day 7 of lactation there was a 2.4-fold increase in serum FGF-21 vs baseline (P < .0001), a 8-fold increase in hepatic FGF-21 mRNA (P < .03), a 2-fold increase in undercarboxylated osteocalcin (Glu13 OCn) (P < .01), and enhanced insulin sensitivity. Recovery of total areal bone density was noted at day 21 of lactation, whereas the femoral trabecular bone volume fraction was still reduced (P < .01). Because FGF-21 levels rose rapidly at day 7 of lactation in B6 lactating mice, we next examined lactating mice with a deletion in the Fgf21 gene. Trabecular and cortical bone masses were maintained throughout lactation in FGF-21(-/-) mice, and pup growth was normal. Compared with lactating control mice, lactating FGF-21(-/-) mice exhibited an increase in bone formation, but no change in bone resorption. In conclusion, in addition to changes in calciotropic hormones, systemic FGF-21 plays a

  7. Relationship between proteolysis in the silo and efficiency of utilization of dietary protein by lactating dairy cows

    USDA-ARS?s Scientific Manuscript database

    Ensiling is used widely to conserve forages for feeding to dairy cows. However, the protein in hay-crop silages is particularly susceptible to microbial breakdown in the rumen, and utilization of protein in alfalfa and grass silages by dairy cows is particularly poor. Dependent on maturity, hay-crop...

  8. Rumen epithelial adaptation to ruminal acidosis in lactating cattle involves the coordinated expression of insulin-like growth factor-binding proteins and a cholesterolgenic enzyme.

    PubMed

    Steele, M A; Dionissopoulos, L; AlZahal, O; Doelman, J; McBride, B W

    2012-01-01

    The objective of this study was to characterize the mRNA expression of metabolic and proliferative genes in the rumen epithelium during ruminal acidosis. To meet our objectives, 16 rumen-fistulated, lactating Holstein dairy cattle (618±35 kg of body weight, 221±32 d in milk) were used in a randomized complete block design. All cattle were fed a high-forage diet (HF; 88.9% of dry matter) for 5 wk before the experiment. After the baseline week (wk 0), half of the cattle were randomly assigned and transitioned to a high-concentrate diet (HC; 62.2% of dry matter) which was fed for 3 wk (wk 1, 2, and 3). For the last 48 h of each week, continuous ruminal pH, short-chain fatty acids, and plasma β-hydroxybutyrate were assessed, followed by a rumen papillae biopsy. Milk production was higher in HC cattle compared with HF during wk 1, 2, and 3 (17.4±0.5 vs. 23.4±0.9 kg/d, respectively); however, the mean ruminal pH was decreased (5.75±0.03 vs. 6.30±0.02). The HC cattle spent more time below pH 5.6 (594±54 vs. 3±3 min/d) and displayed greater concentrations of ruminal butyrate (15.8±0.9 vs. 10.2±0.4 mmol) and plasma β-hydroxybutyrate (1,036±63 vs. 778±20 μM) compared with the HF cattle. The mRNA expression of genes involved in ketogenesis (HMGCS2 and PPARA) and short-chain fatty acid transport (MCT1) was unchanged by treatment. However, a downregulation in HMGCS1 (0.72±0.09), one of the cholesterol biosynthesis genes, was observed in HC cattle during wk 1 of the grain challenge. In addition, the relative mRNA expression value of insulin-like growth factor-binding protein 3 was lower (0.78±0.06), whereas insulin-like growth factor-binding protein 5 was higher (1.79±0.15) in HC compared with HF cattle. These results suggest that grain-induced ruminal acidosis alters the mRNA expression of IGF-binding proteins and a cholesterolgenic enzyme in the rumen epithelium of lactating dairy cattle. Copyright © 2012 American Dairy Science Association. Published by

  9. Effect of amino acids supply in reduced crude protein diets on performance, efficiency of mammary uptake, and transporter gene expression in lactating sows.

    PubMed

    Manjarin, R; Zamora, V; Wu, G; Steibel, J P; Kirkwood, R N; Taylor, N P; Wils-Plotz, E; Trifilo, K; Trottier, N L

    2012-09-01

    To test the hypothesis that reduction in dietary CP concentration coupled with crystalline AA inclusion increases the efficiency of AA use for milk production, mammary AA arteriovenous concentration differences (A-V), AA transport efficiency (A-V/A × 100), and transcript abundance of AA transporters and milk protein genes were determined in lactating sows fed 1 of 3 diets containing 9.5% (Deficient), 13.5% (Ideal), or 17.5% (Standard) CP, with a similar profile of indispensable and dispensable AA. On d 7 and 18, arterial and mammary venous blood and mammary tissue were sampled postfeeding. Transcript abundance of AA transporters b(0,+)AT (SLC7A9), y(+)LAT2 (SLC7A6), ATB(0,+) (SLC6A14), CAT-1 (SLC7A1), and CAT-2b (SLC7A2) and milk protein β-casein (CSN2) and LALBA (α-lactalbumin) were determined using reverse transcription quantitative PCR. Piglet ADG increased curvilinearly (linear and quadratic, P < 0.03) with increasing percent CP from Deficient to Standard. On d 7, Lys and Arg A-V and transport efficiency increased quadratically (P < 0.05) with increasing percent CP. On d 18, Lys A-V tended to increase (linear, P = 0.08) with increasing percent CP. Increasing CP increased Ile and Val A-V on d 7 (linear, P = 0.05 and P = 0.08, respectively) and Leu and Val on d 18 (linear, P = 0.07 and P = 0.04, respectively). On d 7, plasma concentrations of branched chain AA (BCAA):Lys decreased quadratically (P < 0.05). Expression of genes SLC7A9, SLC7A6, SLC6A14, SLC7A1, SLC7A2, CSN2, and LALBA was unaffected by diet. In conclusion, decreasing the dietary CP from 17.5% to 13.5% with inclusion of crystalline AA did not affect piglet ADG, AA transporter, or milk protein gene expression but increased mammary transport efficiency and A-V of Lys and Arg on d 7 of lactation. This increase was associated with a decrease in plasma concentration of BCAA:Lys, suggesting a competitive mechanism between cationic and BCAA for transport of AA across mammary cells.

  10. Short-term administration of rhGH increases markers of cellular proliferation but not milk protein gene expression in normal lactating women

    PubMed Central

    Maningat, Patricia D.; Sen, Partha; Rijnkels, Monique; Hadsell, Darryl L.; Bray, Molly S.

    2011-01-01

    Growth hormone is one of few pharmacologic agents known to augment milk production in humans. We hypothesized that recombinant human GH (rhGH) increases the expression of cell proliferation and milk protein synthesis genes. Sequential milk and blood samples collected over four days were obtained from five normal lactating women. Following 24 h of baseline milk and blood sampling, rhGH (0.1 mg/kg/day) was administered subcutaneously once daily for 3 days. Gene expression changes were determined by microarray studies utilizing milk fat globule RNA isolated from each milk sample. Following rhGH administration, DNA synthesis and cell cycle genes were induced, while no significant changes were observed in the expression of milk synthesis genes. Expression of glycolysis and citric acid cycle genes were increased by day 4 compared with day 1, while lipid synthesis genes displayed a circadian-like pattern. Cell cycle gene upregulation occurred after a lag of ∼2 days, likely explaining the failure to increase milk production after only 3 days of rhGH treatment. We conclude that rhGH induces expression of cellular proliferation and metabolism genes but does not induce milk protein gene expression, as potential mechanisms for increasing milk production and could account for the known effect of rhGH to increase milk production following 7–10 days. PMID:21205870

  11. Regulation of Articular Chondrocyte Proliferation and Differentiation by Indian Hedgehog and Parathyroid Hormone-related Protein

    PubMed Central

    Chen, Xuesong; Macica, Carolyn; Nasiri, Ali; Broadus, Arthur E.

    2008-01-01

    Objective The chondrocytes of the epiphyseal growth zone are regulated by the Indian hedgehog (Ihh)-parathyroid hormone-related protein (PTHrP) axis. In weight-bearing joints, this growth zone comes to be subdivided by the secondary ossification center into distinct articular and growth cartilage structures. Here, we explored the cells of origin, localization, regulation of expression, and putative functions of Ihh and PTHrP in articular cartilage in the mouse. Methods We assessed Ihh and PTHrP expression in an allelic PTHrP-lacZ knockin mouse and several versions of PTHrP-null mice. Selected joints were unloaded surgically to examine load-induction of PTHrP and Ihh. Results The embryonic growth zone appears to serve as the source of PTHrP-expressing proliferative chondrocytes that populate both the forming articular cartilage and growth plate structures. In articular cartilage, these cells take the form of articular chondrocytes in the mid-zone. In PTHrP-knockout mice, mineralizing chondrocytes encroach upon developing articular cartilage but appear to be prevented from mineralizing the joint space by Ihh-driven surface chondrocyte proliferation. In growing and adult mice, PTHrP expression in articular chondrocytes is load-induced, and unloading is associated with rapid changes in PTHrP expression and articular chondrocyte differentiation. Conclusion We conclude that the PTHrP-Ihh axis participates in the maintenance of articular cartilage. Dysregulation of this system might contribute to the pathogenesis of arthritis. PMID:19035497

  12. Genetics Home Reference: lactate dehydrogenase deficiency

    MedlinePlus

    ... dehydrogenase-B pieces (subunits) of the lactate dehydrogenase enzyme. This enzyme is found throughout the body and is important ... cells. There are five different forms of this enzyme, each made up of four protein subunits. Various ...

  13. Dietary carbohydrate composition modifies the milk N efficiency in late lactation cows fed low crude protein diets.

    PubMed

    Cantalapiedra-Hijar, G; Peyraud, J L; Lemosquet, S; Molina-Alcaide, E; Boudra, H; Nozière, P; Ortigues-Marty, I

    2014-02-01

    Nitrogen emissions from dairy cows can be readily decreased by lowering the dietary CP concentration. The main objective of this work was to test whether the milk protein yield reduction associated with low N intakes could be partially compensated for by modifying the dietary carbohydrate composition (CHO). The effects of CHO on digestion, milk N efficiency (milk N/N intake; MNE) and animal performance were studied in four Jersey cows fed 100% or 80% of the recommended protein requirements using a 4×4 Latin square design. Four iso-energetic diets were formulated to two different CHO sources (starch diets with starch content of 34.3% and NDF at 32.5%, and fiber diets with starch content of 5.5% and NDF at 49.1%) and two CP levels (Low=12.0% and Normal=16.5%). The apparent digestible organic matter intake (DOMI) and the protein supply (protein digestible in the small intestine; PDIE) were similar between starch and fiber diets. As planned, microbial N flow (MNF) to the duodenum, estimated from the urinary purine derivatives (PD) excretion, was similar between Low and Normal CP diets. However, the MNF and the efficiency of microbial synthesis (g of microbial N/kg apparently DOMI) were higher for starch v. fiber diets. Milk and milk N fractions (CP, true protein, non-protein N (NPN)) yield were higher for starch compared with fiber diets and for Normal v. Low CP diets. Fecal N excretion was similar across dietary treatments. Despite a higher milk N ouput with starch v. fiber diets, the CHO modified neither the urinary N excretion nor the milk urea-N (MUN) concentration. The milk protein yield relative to both N and PDIE intakes was improved with starch compared with fiber diets. Concentrations of β-hydroxybutyrate, urea and Glu increased and those of glucose and Ala decreased in plasma of cows fed starch v. fiber diets. On the other hand, plasma concentration of albumin, urea, insulin and His increased in cows fed Normal compared with Low CP diets. This study showed

  14. Feeding dried distillers grains with solubles to lactating beef cows: impact of excess protein and fat on cow performance, milk production and pre-weaning progeny growth.

    PubMed

    Shee, C N; Lemenager, R P; Schoonmaker, J P

    2016-01-01

    Multiparous Angus×Simmental cows (n=54, 5.22±2.51 years) with male progeny were fed one of two diets supplemented with either dried distillers grains with solubles (DDGS) or soybean meal (CON), from calving until day 129 postpartum (PP) to determine effects of excess protein and fat on cow performance, milk composition and calf growth. Diets were formulated to be isocaloric and consisted of rye hay and DDGS (19.4% CP; 8.76% fat), or corn silage, rye hay and soybean meal (11.7% CP; 2.06% fat). Cow-calf pairs were allotted by cow and calf age, BW and breed. Cow BW and body condition score (BCS; P⩾0.13) were similar throughout the experiment. A weigh-suckle-weigh was performed on day 64 and day 110±10 PP to determine milk production. Milk was collected on day 68 and day 116±10 PP for analysis of milk components. Milk production was unaffected (P⩾0.75) by dietary treatments. Milk urea nitrogen was increased at both time points in DDGS compared with CON cows (P<0.01). Protein was decreased (P=0.01) and fat was increased (P=0.01) in milk from DDGS compared with CON cows on day 68 PP. Compared to CON, DDGS decreased medium chain FA (P<0.01) and increased long chain FA (P<0.01) at both time points. Saturated FA content of milk was decreased (P<0.01) at both time-points in DDGS compared with CON cows, which resulted in an increase (P<0.01) in monounsaturated and polyunsaturated FA, including cis-9, trans-11 conjugated linoleic acid. Daily gain of the DDGS calves was increased (P=0.01) compared with CON calves, resulting in heavier BW on day 129 (P=0.01). Heavier BW of DDGS calves was maintained through weaning (P=0.01). Timed-artificial insemination (TAI) rates were greater for cows fed DDGS compared with cows fed CON (P<0.02), but dietary treatment had no effect on overall pregnancy rates (P=0.64). In summary, feeding DDGS to lactating beef cows did not change cow BW or BCS, but did improve TAI rates and altered milk composition compared with CON. As a result, male

  15. Effects of crude protein level in concentrate supplements on animal performance and nitrogen utilization of lactating dairy cows fed fresh-cut perennial grass.

    PubMed

    Hynes, D N; Stergiadis, S; Gordon, A; Yan, T

    2016-10-01

    Nitrogen pollution of air and ground water from grazing cattle is of increasing concern. Although several studies have investigated mitigation strategies for nitrogen output from dairy cows fed conserved forages and concentrates, similar research on fresh-cut grass in addition to production parameters is limited. The current study, using 3dietary treatments and incorporating 2 genotypes, was designed to evaluate the effects of concentrate crude protein (CP) levels on animal production and nitrogen utilization efficiency (NUE) in lactating dairy cows. Twelve multiparous cows (6 Holstein and 6 Holstein × Swedish Red) were used in a changeover study with three 25-d periods and 3 diet treatments. Low, medium and high CP concentrate [14.1, 16.1, and 18.1%, respectively, dry matter (DM) basis] diets were fed at 32.8% DM intake combined with good-quality zero-grazed perennial ryegrass (18.2% CP, DM basis). Each period consisted of an adaptation phase (18d) housed as a single group, a 1-d adaptation phase in individual stalls, and a 6-d measurement phase with feed intake and feces, urine, and milk output recorded. We observed no significant interaction between cow genotype and concentrate CP level on any animal performance or NUE parameter. Total DM intake, milk yield and composition, and NUE were not affected by dietary treatment. However, increasing concentrate CP level increased (1) N intake by 42g/d and excretion in urine and manure by 38 and 40g/d, respectively, and (2) the ratio of urine N over manure N. Feeding high CP rather than low CP concentrate increased milk urea N (MUN) content by 3.6mg/dL and total MUN output by 1.08g/d. Crossbred cows had lower grass DM intake, total DM intake, total N intake, and energy-corrected milk yield. However, cow genotype had no significant effect on NUE or MUN parameters. Equations have been developed to predict urine N excretion using MUN output as a sole predictor or in combination with dietary CP level. The present study

  16. Evaluation of Markers of Bone Turnover During Lactation in African-Americans: A Comparison With Caucasian Lactation

    PubMed Central

    Carneiro, Raquel M.; Prebehalla, Linda; Tedesco, Mary Beth; Sereika, Susan M.; Gundberg, Caren M.; Stewart, Andrew F.

    2013-01-01

    Context: The African-American skeleton is resistant to PTH; whether it is also resistant to PTHrP and the hormonal milieu of lactation is unknown. Objectives: The objective of the study was to assess bone turnover markers in African-Americans during lactation vs Caucasians. Design and Participants: A prospective cohort study with repeated measures of markers of bone turnover in 60 African-American women (3 groups of 20: lactating, bottle feeding, and healthy controls), compared with historic Caucasian women. Setting: The study was conducted at a university medical center. Outcome Measures: Biochemical markers of bone turnover and calcium metabolism were measured. Results: 25-Hydroxyvitamin D (25-OHD) and PTH were similar among all 3 African-American groups, but 25-OHD was 30%–50% lower and PTH 2-fold higher compared with Caucasians (P < .001, P < .002), with similar 1,25 dihydroxyvitamin D [1,25(OH)2D] values. Formation markers [amino-terminal telopeptide of procollagen-1 (P1NP) and bone-specific alkaline phosphatase (BSAP)] increased significantly (2- to 3-fold) in lactating and bottle-feeding African-American women (P1NP, P < .001; BSAP, P < .001), as did resorption [carboxy-terminal telopeptide of collagen-1 (CTX) and serum amino-terminal telopeptide of collagen 1 (NTX), both P < .001]. P1NP and BSAP were comparable in African-American and Caucasian controls, but CTX and NTX were lower in African-American vs Caucasian controls. African-American lactating mothers displayed quantitatively similar increases in markers of bone formation but slightly lower increases in markers of resorption vs Caucasians (P = .036). Conclusions: Despite reported resistance to PTH, lactating African-American women have a significant increase in markers of bone resorption and formation in response the hormonal milieu of lactation. This response is similar to that reported in Caucasian women despite racial differences in 25-OHD and PTH. Whether this is associated with similar bone

  17. Composition of sow milk during lactation.

    PubMed

    Klobasa, F; Werhahn, E; Butler, J E

    1987-05-01

    The composition of sow colostrum and milk was quantitated in 25 sows at 14 time points throughout lactation. All animals belonged to the same experimental herd of German Landrace, farrowed within 4 d, and were of various lactation numbers and various litter sizes. In the first 6 h of lactation colostrum total solids (TS) and protein contents were higher, while fat and lactose contents were lower than in mature milk. Decreased total protein and whey protein contents and concomitantly increased fat and lactose content, with nearly unchanged TS levels, indicate transition from colostrum to mature milk. The high protein content of colostrum was largely due to immunoglobulin (Ig). During the first 6 h, IgG accounts for nearly all the protein in colostrum but plays a decreasing role in sow milk as lactation proceeds. After 2 wk, IgA levels begin to increase and at the end of lactation, IgA constitutes 40% of the total whey protein. No influences of lactation number and litter size on milk composition could be ascertained in this study.

  18. Mandibular coronoid process in parathyroid hormone-related protein-deficient mice shows ectopic cartilage formation accompanied by abnormal bone modeling.

    PubMed

    Shibata, Shunichi; Suda, Naoto; Fukada, Kenji; Ohyama, Kimie; Yamashita, Yasuo; Hammond, Vicki E

    2003-07-01

    Parathyroid hormone-related protein (PTHrP) null mutant mice were analyzed to investigate an additional role for PTHrP in cell differentiation. We found ectopic cartilage formation in the mandibular coronoid process in newborn mice. While many previous studies involving PTHrP gene knockout mouse have shown that the cartilage in various regions becomes smaller, this is the first report showing an "increase" of cartilage volume. Investigations of mandibular growth using normal mice indicated that coronoid secondary cartilage never formed from E 15 to d 4, but small amount of cartilage temporally formed at d 7, and this also applies to PTHrP-wild type mice. Therefore, PTHrP deficiency consequently advanced the secondary cartilage formation, which is a novel role of PTHrP in chondrocyte differentiation. In situ hybridization of matrix proteins showed that this coronoid cartilage had characteristics of the lower hypertrophic cell zone usually present at the site of endochondral bone formation and/or "chondroid bone" occasionally found in distraction osteogenesis. In addition, the coronoid process in the PTHrP-deficient mouse also showed abnormal expansion of bone marrow and an increase in the number of multinucleated osteoclasts, an indication of abnormal bone modeling. These results indicate that PTHrP is involved in bone modeling as well as in chondrocyte differentiation. In situ hybridization of matrix protein mRNAs in the abnormal mandibular condylar cartilage revealed that this cartilage was proportionally smaller, supporting previous immunohistochemical results.

  19. Use of different dietary protein sources for lactating goats: milk production and composition as functions of protein degradability and amino acid composition.

    PubMed

    Sanz Sampelayo, M R; Pérez, M L; Gil Extremera, F; Boza, J J; Boza, J

    1999-03-01

    To establish the effect of the nature of four different protein sources [fababeans, 27.8% crude protein (CP); sunflower meal, 41.7% CP; corn gluten feed, 18.8% CP; and cottonseed, 18.3% CP] on milk protein production by goats, the ruminal degradation of these feeds was studied as was the amino acid (AA) composition of the original material and that of the undegradable fractions of the protein sources. Four diets were designed; 20% of their protein was supplied by each of the different sources. Four groups of 5 Granadina goats were used to study the utilization of these diets for milk production. No significant differences were observed in dry matter intake or milk production. The milk produced by goats fed the diet containing sunflower meal had the lowest protein concentration; the highest milk protein concentration was observed for goats fed the diet containing corn gluten feed. From a multivariate analysis, it was deduced that the quickly degradable protein fraction in the rumen and the ruminally undegradable protein fraction were the components of the protein sources most directly related to the milk protein produced. Given the similar AA profiles of the undegradable fractions of the different protein sources, the possible supplementation achieved from these ruminally undegradable fractions must be established by the amount of protein supplied regardless of AA composition.

  20. Regulation of human organic anion transporter 4 by parathyroid hormone-related protein and protein kinase A

    PubMed Central

    Duan, Peng; Li, Shanshan; You, Guofeng

    2012-01-01

    Human organic anion transporter 4 (hOAT4) belongs to a family of organic anion transporters that play critical roles in the body disposition of clinically important drugs, including anti-human immunodeficiency virus therapeutics, anti-tumor drugs, antibiotics, antihypertensives, and anti-inflammatories. hOAT4 is abundantly expressed in the kidney and placenta. In the current study, we examined the regulation of hOAT4 by parathyroid hormone-related protein (PTHrP) and protein kinase A (PKA) in kidney COS-7 cells. PTHrP induced a time- and concentration-dependent stimulation of hOAT4 transport activity. The stimulation of hOAT4 activity by PTHrP mainly resulted from an increased cell surface expression without a change in total cell expression of the transporter. Activation of PKA by Bt2-cAMP also resulted in a stimulation of hOAT4 activity through an increased cell surface expression of the transporter. However, PTHrP-induced stimulation of hOAT4 activity could not be prevented by treating hOAT4-expressing cells with the PKA inhibitor H89. We concluded that both PTHrP and activation of PKA stimulate hOAT4 activity through redistribution of the transporter from intracellular compartments to the cell surface. However, PTHrP regulates hOAT4 activity by mechanisms independent of PKA pathway. PMID:23097748

  1. Incompatibility of silver nanoparticles with lactate dehydrogenase leakage assay for cellular viability test is attributed to protein binding and reactive oxygen species generation.

    PubMed

    Oh, Seok-Jeong; Kim, Hwa; Liu, Yingqiu; Han, Hyo-Kyung; Kwon, Kyenghee; Chang, Kyung-Hwa; Park, Kwangsik; Kim, Younghun; Shim, Kyuhwan; An, Seong Soo A; Lee, Moo-Yeol

    2014-03-21

    A growing number of studies report that conventional cytotoxicity assays are incompatible with certain nanoparticles (NPs) due to artifacts caused by the distinctive characteristics of NPs. Lactate dehydrogenase (LDH) leakage assays have inadequately detected cytotoxicity of silver nanoparticles (AgNPs), leading to research into the underlying mechanism. When ECV304 endothelial-like umbilical cells were treated with citrate-capped AgNPs (cAgNPs) or bare AgNPs (bAgNPs), the plasma membrane was disrupted, but the LDH leakage assay failed to detect cytotoxicity, indicating interference with the assay by AgNPs. Both cAgNPs and bAgNPs inactivated LDH directly when treated to cell lysate as expected. AgNPs adsorbed LDH and thus LDH, together with AgNPs, was removed from assay reactants during sample preparation, with a resultant underestimation of LDH leakage from cells. cAgNPs, but not bAgNPs, generated reactive oxygen species (ROS), which were successfully scavenged by N-acetylcysteine or ascorbic acid. LDH inhibition by cAgNPs could be restored partially by simultaneous treatment with those antioxidants, suggesting the contribution of ROS to LDH inactivation. Additionally, the composition of the protein corona surrounding AgNPs was identified employing liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. In sum, the LDH leakage assay, a conventional cell viability test method, should be employed with caution when assessing cytotoxicity of AgNPs.

  2. Effects of sodium lactate Ringer's injection on transfection of human protein kinase C-α antisense oligonucleotide in A549 lung cancer cells.

    PubMed

    Wang, Z H; Sun, W W; Han, Y L; Ma, Z

    2016-08-26

    In the present study, we evaluated the effects of four solutions [Dulbecco's modified Eagle's medium (DMEM), sodium lactate Ringer's injection (SLRI), phosphate-buffered saline (PBS), and NaCl] on the transfection of the human protein kinase C-a antisense oligonucleotide (PKC-a ASO) aprinocarsen in human lung carcinoma A549 cells. Specifically, SLRI, DMEM, PBS, or NaCl were used as the growth solutions for A549 cells, and OPTI-MEM was used as the PKC-a ASO diluent for transfection. Additionally, SLRI, DMEM, PBS, or NaCl were used as both the growth solutions and diluents for transfection. The cell viability and transfection efficiency were determined. The results demonstrated that when SLRI was used as either the growth solution or both the growth solution and diluent for aprinocarsen transfection in A549 cells, the effects were close to the best effects observed with DMEM as the growth solution and OPTI-MEM as the diluent, which supported the transfection of aprinocarsen into the cells. Moreover, SLRI resulted in higher transfection efficiency than those of PBS and NaCl. In in vitro experiments, aprinocarsen effectively induced apoptosis in A549 cells. In conclusion, SLRI may replace PBS or NaCl in clinical trials as a transfection solution readily accepted by the human body. To our knowledge, this is the first report demonstrating the use of SLRI as a transfection solution in lung-cancer cell lines.

  3. Effects of concentrate crude protein content on nutrient digestibility, energy utilization, and methane emissions in lactating dairy cows fed fresh-cut perennial grass.

    PubMed

    Hynes, D N; Stergiadis, S; Gordon, A; Yan, T

    2016-11-01

    Although many studies have investigated mitigation strategies for methane (CH4) output from dairy cows fed a wide variety of diets, research on the effects of concentrate crude protein (CP) content on CH4 emissions from dairy cows offered fresh grass is limited. The present study was designed to evaluate the effects of cow genotype and concentrate CP level on nutrient digestibility, energy utilization, and CH4 emissions in dairy cows offered fresh-grass diets. Twelve multiparous lactating dairy cows (6 Holstein and 6 Holstein × Swedish Red) were blocked into 3 groups for each breed and assigned to a low-, medium-, or high-CP concentrate diet [14.1, 16.1, and 18.1% CP on a dry matter (DM) basis, respectively], in a 3-period changeover study (25d per period). Total diets contained (DM basis) 32.8% concentrates and 67.2% perennial ryegrass, which was harvested daily. All measurements were undertaken during the final 6d of each period: digestibility measurements for 6d and calorimetric measurements in respiration chambers for 3d. Feed intake and milk production data were reported in a previous paper. We observed no significant interaction between concentrate CP level and cow genotype on any parameter. Concentrate CP level had no significant effect on any energy utilization parameter, except for urinary energy output, which was positively related to concentrate CP level. Similarly, concentrate CP content had no effect on CH4 emission (g/d), CH4 per kg feed intake, or nutrient digestibility. Cross breeding of Holstein cows significantly reduced gross energy, digestible energy, and metabolizable energy intake, heat production, and milk energy output. However, cow genotype had no significant effect on energy utilization efficiency or CH4 parameters. Furthermore, the present study yielded a value for gross energy lost as CH4 (5.6%) on fresh grass-based diets that was lower than the widely accepted value of 6.5%. The present findings indicate that reducing concentrate CP

  4. Development of nitrogen and methane losses in the first eight weeks of lactation in Holstein cows subjected to deficiency of utilisable crude protein under restrictive feeding conditions.

    PubMed

    Sutter, Franz; Schwarm, Angela; Kreuzer, Michael

    2017-02-01

    Low-protein diets are increasingly being used in dairy cow nutrition to minimise noxious nitrogen (N) emissions. However, at parturition, the lower milk yield at that time may mask deficiency in dietary utilisable crude protein (uCP; equivalent to metabolisable protein). Under restrictive feeding conditions, farmers would limit the feed allowance to match the lower measured milk yield, thereby exacerbating the deficiency. The consequences for N emission intensity per kg milk yield and methane emissions are unknown. In this study, two diets were fed to nine Holstein cows each from parturition onwards. One diet was complete and the other was calculated as 20% deficient in uCP. Feed allowance was always oriented towards the measured milk yield. In each of the first eight lactation weeks, intake and excretion were measured for 5 d. On the last 2 d of this period, methane emission was measured in respiration chambers. The statistical model included treatment, week and interaction as effects. The real levels of uCP and energy supply across the 8 weeks were 33% and 15% below requirements, respectively, in the Deficient cows. In addition, the Deficient cows consumed 18% less dry matter (caused by substantial refusals in week 1, where energy supply was according to requirements) and produced 25% less milk (26 vs. 34 kg/d). Cows in both groups used dietary N with similar efficiency for milk protein synthesis and excreted similar proportions of the N ingested via urine and faeces. This resulted in both treatments having similar N emission intensities per kg milk N and similar urinary N as a proportion of total excreta N, suggesting a similar potential for gaseous N emissions from the manure per kg of milk. The Deficient cows emitted 22% less methane overall but had similar methane yield and emission intensity to the Controls. In conclusion, a reduction in crude protein intake immediately after parturition does not reduce N emission per unit of milk when associated with u

  5. Effects of low rumen-degradable protein or abomasal fructan infusion on diet digestibility and urinary nitrogen excretion in lactating dairy cows.

    PubMed

    Gressley, T F; Armentano, L E

    2007-03-01

    Post-ileal carbohydrate fermentation in dairy cows converts blood urea nitrogen (BUN) into fecal microbial protein. This should reduce urinary N, increase fecal N, and reduce manure NH3 volatilization. However, if intestinal BUN recycling competes with ruminal BUN recycling, hindgut fermentation may reduce NH3 for rumen microbial protein synthesis. Eight lactating Holstein cows were used in a replicated 4 x 4 Latin square design with 14-d periods. Treatments were arranged as a 2 x 2 factorial. Diets contained either adequate rumen-degradable protein (RDP; high RDP) or were 28% below predicted RDP requirements (low RDP). Cows received abomasal infusions of either 10 L/d of saline or 10 L/d of saline containing 1 kg/d of inulin. We hypothesized that reducing ruminal NH3, either by restricting RDP intake or by diverting BUN to feces with inulin, would reduce rumen microbial protein synthesis, as would be evidenced by significant main effects of treatments on rumen NH3, milk production, and urinary purine derivative excretion. Furthermore, we thought it likely that effects of inulin might be greater when rumen NH3 was already low, as would be indicated by significant interactions between inulin infusion and dietary RDP level on rumen NH3, milk production, and urinary purine derivative excretion. Rumen NH3 was reduced by the low-RDP diet, but urinary purine derivative excretion and milk production were unaffected. However, the low-RDP diet reduced apparent total tract digestibility of OM and starch and reduced in situ rumen NDF digestibility. Abomasal inulin reduced the BUN concentration but did not affect milk yield or rumen NH3, suggesting that RDP requirements are not affected by hindgut fermentation. Inulin shifted 23 g/d of N from urine to feces. However, based on fecal purine excretion, we estimated that only 8 g/d of the increased fecal N was due to increased fecal microbial output. Inulin reduced true digestibility of dietary protein or increased nonmicrobial as

  6. Role of parathyroid hormone-related protein in the pro-inflammatory and pro-fibrogenic response associated with acute pancreatitis.

    PubMed

    Bhatia, Vandanajay; Kim, Sung O K; Aronson, Judith F; Chao, Celia; Hellmich, Mark R; Falzon, Miriam

    2012-04-10

    Pancreatitis is a common and potentially lethal necro-inflammatory disease with both acute and chronic manifestations. Current evidence suggests that the accumulated damage incurred during repeated bouts of acute pancreatitis (AP) can lead to chronic disease, which is associated with an increased risk of pancreatic cancer. While parathyroid hormone-related protein (PTHrP) exerts multiple effects in normal physiology and disease states, its function in pancreatitis has not been previously addressed. Here we show that PTHrP levels are transiently elevated in a mouse model of cerulein-induced AP. Treatment with alcohol, a risk factor for both AP and chronic pancreatitis (CP), also increases PTHrP levels. These effects of cerulein and ethanol are evident in isolated primary acinar and stellate cells, as well as in the immortalized acinar and stellate cell lines AR42J and irPSCc3, respectively. Ethanol sensitizes acinar and stellate cells to the PTHrP-modulating effects of cerulein. Treatment of acinar cells with PTHrP (1-36) increases expression of the inflammatory mediators interleukin-6 (IL-6) and intracellular adhesion protein (ICAM-1), suggesting a potential autocrine loop. PTHrP also increases apoptosis in AR42J cells. Stellate cells mediate the fibrogenic response associated with pancreatitis; PTHrP (1-36) increases procollagen I and fibronectin mRNA levels in both primary and immortalized stellate cells. The effects of cerulein and ethanol on levels of IL-6 and procollagen I are suppressed by the PTH1R antagonist, PTHrP (7-34). Together these studies identify PTHrP as a potential mediator of the inflammatory and fibrogenic responses associated with alcoholic pancreatitis.

  7. Parathyroid hormone-related protein inhibits DKK1 expression through c-Jun-mediated inhibition of β-Catenin activation of the DKK1 promoter in prostate cancer

    PubMed Central

    Zhang, H.; Yu, C.; Dai, J.; Keller, JM.; Hua, A.; Sottnik, JL.; Shelley, G.; Hall, CL.; Park, SI.; Yao, Z.; Zhang, J.; McCauley, LK.; Keller, ET.

    2014-01-01

    Prostate cancer bone metastases are unique in that that majority of them induce excessive mineralized bone matrix, through undefined mechanisms, as opposed to most other cancers that induce bone resorption. Parathyroid hormone-related protein (PTHrP) is produced by prostate cancer cells and intermittent PTHrP exposure has bone anabolic effects suggesting PTHrP could contribute to the excess bone mineralization. Wnts are bone productive factors produced by prostate cancer cells and the Wnt inhibitor DKK1 has been shown to promote prostate cancer progression. These findings, in conjunction with the observation that PTHrP expression increases and DKK1 expression decreases as prostate cancer progresses led to the hypothesis that PTHrP could be a negative regulator of DKK1 expression in prostate cancer cells, and hence allow the osteoblastic activity of Wnts to be realized. To test this, we first demonstrated that PTHrP downregulated DKK1 mRNA and protein expression. We then found through multiple mutated DKK1 promoter assays that PTHrP, through c-Jun activation, downregulated the DKK1 promoter through a TCF-response element site. Furthermore, chromatin immunoprecipitation (ChIP) and reChIP assays revealed that PTHrP-mediated this effect through inducing c-Jun to bind to a transcriptional activator complex consisting of β-catenin that binds the most proximal DKK1 promoter TCF-response element. Together, these results demonstrate a novel signaling linkage between PTHrP and Wnt signaling pathways that results in downregulation of a Wnt inhibitor allowing for Wnt activity that could contribute the osteoblastic nature of prostate cancer. PMID:23752183

  8. Influence of protein supplementation during late pregnancy and lactation on the resistance of Santa Ines and Ile de France ewes to Haemonchus contortus.

    PubMed

    Rocha, R A; Bricarello, P A; Silva, M B; Houdijk, J G M; Almeida, F A; Cardia, D F F; Amarante, A F T

    2011-09-27

    This experiment aimed to evaluate the effect of periparturient metabolizable protein (MP) nutrition on resistance to Haemonchus contortus in single rearing Ile de France and Santa Ines ewes. The restriction-fed iso-energetic diet was calculated to provide either 0.8 (low MP diet) or 1.3 (high MP diet) times MP, from three weeks before parturition until eight weeks into lactation. The ewes were experimentally infected with 1000 H. contortus infective larvae (L3) three times a week (Mondays, Wednesdays and Fridays), starting five weeks before the predicted date for parturition until a total of 15,000 L3 had been administered. While both breeds showed elevated fecal egg counts (FEC), these values were significantly lower for Santa Ines ewes than Ile de France ewes, but were independent of level of MP feeding. The latter also did not affect lamb weight gain and ewe body weight variation in each breed. Packed cell volume and total plasma protein for Santa Ines in all periods were significantly higher than those for Ile de France ewes (P<0.01) but were not affected by nutrition. In contrast, levels of serum IgG and IgA antibodies against somatic H. contortus infective larvae and adult antigens were similar between breeds but higher in animals that received high MP diets (P<0.05). The reduced body score of ewes at the beginning of the experiment probably influenced their high susceptibility to incoming larvae. Since, unexpectedly, MP scarcity was not achieved in this experiment, our data support the view that Santa Ines ewes are more resistant to H. contortus than Ile de France ewes.

  9. Hedgehog signaling in bone regulates whole-body energy metabolism through a bone-adipose endocrine relay mediated by PTHrP and adiponectin.

    PubMed

    Zhang, Xu; Cheng, Qianni; Wang, Yixiang; Leung, Po Sing; Mak, Kinglun Kingston

    2017-02-01

    Bone plays a role in energy metabolism, but the interplay between bone and other organs in this process is not completely understood. Here, we show that upregulated Hh signaling in bones results in increased whole-body energy expenditure, white adipose tissue (WAT) browning, hypoglycemia and skeletal muscle atrophy. We found that Hh signaling induces PTHrP secretion from bones and causes WAT browning. Injection of PTHrP-neutralizing antibody attenuates WAT browning and improves the circulating blood glucose level while high-fat diet treatment only rescues hypoglycemia. Furthermore, bone-derived PTHrP stimulates adiponectin secretion in WAT and results in systemic increase of fatty acid oxidation and glucose uptake. Mechanistically, PTHrP activates both PKA/cAMP and Akt/Foxo pathways for Ucp1 expression in WAT. PTHrP couples adiponectin actions to activate the AMPK pathway in the skeletal muscles and liver, respectively, for fatty acid oxidation. Our findings establish a new bone-adipose hormonal relay that regulates whole-body energy metabolism.

  10. Lactation and reproduction*

    PubMed Central

    Thomson, A. M.; Hytten, F. E.; Black, A. E.

    1975-01-01

    The authors review the literature on the effect of lactation on fertility in the absence of contraception and on the effects of contraceptive measures on lactation. They examine data from several countries on the intervals between births and on the return of menstruation and ovulation after childbirth, comparing lactating with nonlactating women. They conclude that lactation is an inefficient contraceptive for the individual, but that in populations sustained lactation is associated with reduced fertility. Possible physiological mechanisms causing lactation amenorrhoea are discussed. Though much of the literature on the effect of contraceptives on lactation is inadequate, there is general agreement that the estrogen component of hormonal preparations has an adverse effect on lactation, but that progestins alone do not. Many questions remain. Is this effect seen in established lactation, or only in the puerperal period? Is it a direct pharmacological effect, or are pill-users the mothers least motivated to maintain breast-feeding? Does a close relationship exist between hormones given and lactation performance? The authors comment on some of the technical deficiencies of previous studies in this field and discuss practical possibilities of, and limitations to, obtaining adequate scientific information in the future. PMID:1084804

  11. Ratio of dietary rumen degradable protein to rumen undegradable protein affects nitrogen partitioning but does not affect the bovine milk proteome produced by mid-lactation Holstein dairy cows.

    PubMed

    Tacoma, R; Fields, J; Ebenstein, D B; Lam, Y-W; Greenwood, S L

    2017-09-01

    Little is known about the bovine milk proteome or whether it can be affected by diet. The objective of this study was to determine if the dietary rumen degradable protein (RDP):rumen undegradable protein (RUP) ratio could alter the bovine milk proteome. Six Holstein cows (parity: 2.5 ± 0.8) in mid lactation were blocked by days in milk (80 ± 43 d in milk) and milk yield (57.5 ± 6.0 kg) and randomly assigned to treatment groups. The experiment was conducted as a double-crossover design consisting of three 21-d periods. Within each period, treatment groups received diets with either (1) a high RDP:RUP ratio (RDP treatment: 62.4:37.6% of crude protein) or (2) a low RDP:RUP ratio (RUP treatment: 51.3:48.7% of crude protein). Both diets were isonitrogenous and isoenergetic (crude protein: 18.5%, net energy for lactation: 1.8 Mcal/kg of dry matter). To confirm N and energy status of cows, dry matter intake was determined daily, rumen fluid samples were collected for volatile fatty acid analysis, blood samples were collected for plasma glucose, β-hydroxybutyrate, urea nitrogen, and fatty acid analysis, and total 24-h urine and fecal samples were collected for N analysis. Milk samples were collected to determine the general milk composition and the protein profile. Milk samples collected for high-abundance protein analysis were subjected to HPLC analysis to determine the content of α-casein, β-casein, and κ-casein, as well as α-lactalbumin and β-lactoglobulin. Samples collected for low-abundance protein analysis were fractionated, enriched using ProteoMiner treatment, and separated using sodium dodecyl sulfate-PAGE. After excision and digestion, the peptides were analyzed using liquid chromatography (LC) tandem mass spectrometry (MS/MS). The LC-MS/MS data were analyzed using PROC GLIMMIX of SAS (version 9.4, SAS Institute Inc., Cary, NC) and adjusted using the MULTTEST procedure. All other parameters were analyzed using PROC MIXED of SAS. No treatment differences

  12. Determination of total proteins and lactate dehydrogenase for the diagnosis of pleural transudates and exudates: redefining the classical criterion with a new statistical approach.

    PubMed

    Maranhão, Bernardo Henrique Ferraz; Silva Junior, Cyro Teixeira da; Chibante, Antonio Monteiro da Silva; Cardoso, Gilberto Perez

    2010-01-01

    To propose a new classification criterion for the differentiation between pleural exudates and transudates-quantifying total proteins in pleural fluid (TP-PF) and lactate dehydrogenase in pleural fluid (LDH-PF) exclusively-as well as to compare this new criterion with the classical criterion in terms of diagnostic yield. This was an observational, cross-sectional study with a within-subject design, comprising 181 patients with pleural effusion treated at two university hospitals in the state of Rio de Janeiro, Brazil, between 2003 and 2006. The diagnostic parameters included in the classical criterion were identified, as were those included in the new criterion. Of the 181 patients, 152 and 29 were diagnosed with pleural exudates and pleural transudates, respectively. For the classical criterion, the sensitivity, specificity, and accuracy for the diagnosis of pleural exudates were, respectively, 99.8%, 68.6%, and 94.5%, whereas the corresponding values for the diagnosis of pleural transudates were 76.1%, 90.1%, and 87.6%. For the new criterion (cut-off points set at 3.4 g/dL for TP-PF and 328.0 U/L for LDH-PF), the sensitivity, specificity, and accuracy for the diagnosis of exudates were, respectively, 99.4%, 72.6%, and 99.2%, whereas the corresponding values for the diagnosis of transudates were 98.5%, 83.4%, and 90.0%. The accuracy of the new criterion for the diagnosis of pleural exudates was significantly greater than was that of the classical criterion (p = 0.0022). The diagnostic yield was comparable between the two criteria studied. Therefore, the new classification criterion can be used in daily practice.

  13. α-Retinol is distributed through serum retinol-binding protein-independent mechanisms in the lactating sow-nursing piglet dyad.

    PubMed

    Dever, Joseph T; Surles, Rebecca L; Davis, Christopher R; Tanumihardjo, Sherry A

    2011-01-01

    α-Retinol (αR) is a structural isomer of retinol [vitamin A (VA)] that does not bind to serum retinol-binding protein (RBP). In this study, α-retinyl acetate (αRA) was synthesized and given orally (35 μmol) to VA-deficient lactating sows (n = 11) to assess its potential to trace RBP-independent retinol transport and tissue uptake. The αRA dose primarily appeared in sow serum as 4 α-retinyl esters (αRE) with peak serum total αR concentrations (the sum of the alcohol and ester forms) detected at 2 h (70 ± 23 nmol/L, mean ± SEM) postdose. From 0 to 40 h postdose, the percentage of serum total αR in the alcohol form did not increase. Rapid αR uptake into sow milk was observed with peak concentrations (371 ± 83 nmol/L) at 7.5 h postdose, consistent with the uptake of αRE from chylomicra. A high percentage of the αRA dose (62 ± 15%, mean ± SD) was present in the livers of sows (n = 6) killed 22-28 d postdose. Approximately 15-26% of the sow αRA dose was transferred to the livers of the nursing piglets (n = 17) after 3 d. In piglets and sows, a similar percentage of hepatic total αR was detected in the ester form as that of hepatic total retinol. Taken together, these data suggest that an oral dose of αRA effectively traces the uptake, esterification, chylomicron transport, and hepatic storage of retinol and may be useful for deciphering the role of RBP-independent delivery of retinol to other tissues.

  14. Parathyroid hormone-related protein (107-111) improves the bone regeneration potential of gelatin-glutaraldehyde biopolymer-coated hydroxyapatite.

    PubMed

    Lozano, Daniel; Sánchez-Salcedo, Sandra; Portal-Núñez, Sergio; Vila, Mercedes; López-Herradón, Ana; Ardura, Juan Antonio; Mulero, Francisca; Gómez-Barrena, Enrique; Vallet-Regí, María; Esbrit, Pedro

    2014-07-01

    Biopolymer-coated nanocrystalline hydroxyapatite (HA) made as macroporous foams which are degradable and flexible are promising candidates as orthopaedic implants. The C-terminal (107-111) epitope of parathyroid hormone-related protein (PTHrP) exhibits osteogenic properties. The main aim of this study was to evaluate whether PTHrP (107-111) loading into gelatin-glutaraldehyde biopolymer-coated HA (HAGlu) scaffolds would produce an optimal biomaterial for tissue engineering applications. HAGlu scaffolds with and without PTHrP (107-111) were implanted into a cavitary defect performed in both distal tibial metaphysis of adult rats. Animals were sacrificed after 4 weeks for histological, microcomputerized tomography and gene expression analysis of the callus. At this time, bone healing occurred only in the presence of PTHrP (107-111)-containing HAGlu implant, related to an increase in bone volume/tissue volume and trabecular thickness, cortical thickness and gene expression of osteocalcin and vascular cell adhesion molecule 1, but a decreased gene expression of Wnt inhibitors, SOST and dickkopf homolog 1. The autonomous osteogenic effect of the PTHrP (107-111)-loaded HAGlu scaffolds was confirmed in mouse and human osteoblastic cell cultures. Our findings demonstrate the advantage of loading PTHrP (107-111) into degradable HAGlu scaffolds for achieving an optimal biomaterial that is promising for low load bearing clinical applications. Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  15. Current trends in lactate metabolism: introduction.

    PubMed

    Gladden, L Bruce

    2008-03-01

    In September 2006, at the Integrative Physiology of Exercise meeting in Indianapolis, IN, a symposium entitled "Current Trends in Lactate Metabolism" was presented. This short paper introduces two papers from that symposium. The first paper by L. Bruce Gladden briefly summarizes key pieces of evidence that support the cell-to-cell lactate shuttle, a concept that is no longer an hypothesis but that, instead, is now an established theory that provides the context for discussions of whole body metabolism. Gladden also offers a critical appraisal of the intracellular lactate shuttle and evaluates an ongoing controversy relative to the role of lactate in acid-base balance. In the second paper, Hashimoto and Brooks provide their evidence in support of the intracellular lactate shuttle and a lactate oxidation complex in the inner mitochondrial membrane. They also postulate that lactate is a cell-signaling molecule, "lactormone," that can upregulate gene and protein expression. Both papers have been updated since their original presentations and represent the current state of knowledge.

  16. Effect of replacing soybean meal protein with protein from upland cottonseed, Pima cottonseed, or extruded Pima cottonseed on production of lactating dairy cows

    USDA-ARS?s Scientific Manuscript database

    Pima cotton production is increasing in the U.S., but Pima cottonseed contains higher concentrations of the toxic pigment gossypol than conventional upland cottonseed. Heating promotes gossypol reaction with protein, reducing gossypol absorption and toxicity. The objective of this study was to asses...

  17. A low maternal protein diet during pregnancy and lactation has sex- and window of exposure-specific effects on offspring growth and food intake, glucose metabolism and serum leptin in the rat.

    PubMed

    Zambrano, E; Bautista, C J; Deás, M; Martínez-Samayoa, P M; González-Zamorano, M; Ledesma, H; Morales, J; Larrea, F; Nathanielsz, P W

    2006-02-15

    Extensive epidemiological and experimental evidence indicates that a sub-optimal environment during fetal and neonatal development in both humans and animals may programme offspring susceptibility to later development of chronic diseases including obesity and diabetes that are the result of altered carbohydrate metabolism. We determined the effects of protein restriction during pregnancy and/or lactation on growth, serum leptin, and glucose and insulin responses to a glucose tolerance test in male and female offspring at 110 days postnatal life. We fed Wistar rats a normal control 20% casein diet (C) or a restricted diet (R) of 10% casein during pregnancy. Female but not male R pups weighed less than C at birth. After delivery, mothers received the C or R diet during lactation to provide four offspring groups: CC (first letter maternal pregnancy diet and second maternal lactation diet), RR, CR and RC. All offspring were fed ad libitum with C diet after weaning. Relative food intake correlated inversely with weight. Offspring serum leptin correlated with body weight and relative, but not absolute, food intake in both male and female pups. Serum leptin was reduced in RR female pups compared with CC and increased in RC males compared with CC at 110 days of age. Offspring underwent a glucose tolerance test (GTT) at 110 days postnatal life. Female RR and CR offspring showed a lower insulin to glucose ratio than CC. At 110 days of age male RR and CR also showed some evidence of increased insulin sensitivity. Male but not female RC offspring showed evidence of insulin resistance compared with CC. Cholesterol was similar and triglycerides (TG) higher in male compared with female CC. Cholesterol and TG were higher in males than females in RR, CR and RC (P < 0.05). Cholesterol and TG did not differ between groups in females. Cholesterol and TG were elevated in RC compared with CC males. Nutrient restriction in lactation increased relative whole protein and decreased whole

  18. Effects of temperature-humidity index and chromium supplementation on antioxidant capacity, heat shock protein 72, and cytokine responses of lactating cows.

    PubMed

    Zhang, F J; Weng, X G; Wang, J F; Zhou, D; Zhang, W; Zhai, C C; Hou, Y X; Zhu, Y H

    2014-07-01

    Heat stress adversely affects the productivity and immune status of dairy cows. The temperature-humidity index (THI) is commonly used to indicate the degree of heat stress on dairy cattle. We investigated the effects of different THI and Cr supplementation on the antioxidant capacity, the levels of heat shock protein 72 (Hsp72), and cytokine responses of lactating cows. The study used a total of 24 clinically healthy uniparous midlactation Holstein cows, which were randomly divided into 2 groups (n = 12 per group), and was conducted in 3 designated THI periods: low THI period (LTHI; THI = 56.4 ± 2.5), moderate THI period (MTHI; THI = 73.9 ± 1.7), and high THI period (HTHI; THI = 80.3 ± 1.0). The 2 groups of cows were fed corn and corn silage based basal diet supplemented chromium picolinate to provide 3.5 mg of Cr/cow daily (Cr+) or basal diet with no Cr (Cr-). The experiment was a 3 × 2 factorial design. The numbers of leukocytes (P < 0.05) and serum levels of glucose (P < 0.001) were lower; however, the serum levels of blood urea nitrogen (BUN; P < 0.001) and creatinine (P < 0.001) were greater in the MTHI and HTHI than in LTHI. The total antioxidant capacity in the serum was unaltered; an increase in superoxide dismutase activity (P < 0.001) and in serum malondialdehyde concentration (P < 0.001) was observed in the MTHI and HTHI compared with the LTHI. The high THI led to increases in serum concentrations of tumor necrosis factor-α (TNF-α; P < 0.001) and IL-10 (P < 0.05). Cows supplemented with Cr had lower (P = 0.009) serum concentrations of cholesterol but greater (P < 0.001, respectively) serum levels of Hsp72 and IL-10 compared with those without Cr supplementation in the HTHI. Western blot analysis revealed that cows supplemented with Cr had greater (P = 0.038) expression of the inhibitor of nuclear factor kappa B α (IκBα) in peripheral blood mononuclear cells (PBMC) compared with those without Cr supplementation in the HTHI, whereas the expression

  19. Physiology of lactation

    USDA-ARS?s Scientific Manuscript database

    The breast changes in size, shape, and function during puberty, pregnancy, and lactation. The physiology of lactation is reviewed here. The breast is composed of fat and connective tissue that supports a tubuloalveolar structure. During development, anatomic changes involving new lobule formation an...

  20. Effect of dietary protein level and rumen-protected amino acid supplementation on amino acid utilization for milk protein in lactating dairy cows.

    PubMed

    Lee, C; Giallongo, F; Hristov, A N; Lapierre, H; Cassidy, T W; Heyler, K S; Varga, G A; Parys, C

    2015-03-01

    This study investigated the effect of metabolizable protein (MP) supply and rumen-protected (RP) Lys and Met supplementation on productivity, nutrient digestibility, urinary N losses, apparent total-tract digestibility of dietary AA, and the efficiency of AA utilization for milk protein synthesis in dairy cows. The experiment was conducted with 8 ruminally cannulated Holstein cows in a replicated 4×4 Latin square design trial with 21-d periods. Treatments were (1) MP-adequate diet (AMP; MP balance of -24 g/d); (2) MP-deficient diet (DMP; MP balance of -281 g/d); (3) DMP supplemented with 100 g of RPLys/cow per day (estimated digestible Lys supply=24 g/d; DMPL; MP balance of -305g/d); and (4) DMPL supplemented with 24 g of RPMet/cow per day (estimated digestible Met supply=15 g/d; DMPLM; MP balance of -256g/d). Diet had no effect on total-tract nutrient digestibility, milk production, and milk composition, but the DMP diets decreased urinary N excretion and the ammonia emitting potential of manure. Plasma Met concentration was increased by DMPLM compared with AMP. Supplementation with RPLys had no effect on plasma Lys. Concentration of most AA in milk protein was increased or tended to be increased by DMPLM compared with DMPL. Except for the AA supplemented as RPAA (i.e., Met and Lys), apparent total-tract digestibility of all dietary AA was generally greater for the DMP diets and ranged from 33% (Arg, AMP diet) to 67% (Thr, DMPL diet). Apparent recovery of dietary AA in milk protein followed the same trends, being greater for the DMP diets than AMP and generally lower for Lys and Met with the RPAA-supplemented diets versus AMP and DMP. The RPAA were apparently not used for milk protein synthesis in the conditions of this experiment. The AA recoveries in milk protein varied from around 17% (Ala) to 70% (Pro). Milk protein recoveries of essential AA (EAA) were around 54% for the DMP diet and 49% for AMP. The estimated efficiency of utilization of digestible EAA for

  1. Lowering rumen-degradable protein maintained energy-corrected milk yield and improved nitrogen-use efficiency in multiparous lactating dairy cows exposed to heat stress.

    PubMed

    Kaufman, J D; Kassube, K R; Ríus, A G

    2017-10-01

    The objective of this study was to examine the effect of reducing rumen-degradable protein (RDP) and rumen-undegradable protein (RUP) proportions on feed intake, milk production, and N-use efficiency in primiparous and multiparous cows exposed to warm climates. Eighteen primiparous and 30 multiparous mid-lactation Holstein cows were used in a completely randomized design with a 2 × 2 factorial arrangement of treatments. Cows were randomly assigned to 1 of 4 dietary treatments formulated to contain 2 proportions of RDP (10 and 8%) and 2 proportions RUP (8 and 6%) of dry matter (DM) indicated as follows: (1) 10% RDP, 8% RUP; (2) 8% RDP, 8% RUP; (3) 10% RDP, 6% RUP; and (4) 8% RDP, 6% RUP. Protein sources were manipulated to obtain desired RDP and RUP proportions. Diets were isoenergetic and contained 50% forage and 50% concentrate (DM basis). Cows were individually fed the 10% RDP, 8% RUP diet 3 wk before treatment allocation. Cows were exposed to the prevailing Tennessee July and August temperature and humidity in a freestall barn with no supplemental cooling. Main effects and their interaction were tested using the Mixed procedure of SAS (least squares means ± standard error of the mean; SAS Institute Inc., Cary, NC). Observed values of nutrient intake and milk production were used to obtain NRC (2001) model predictions. Cows showed signs of heat stress throughout the study. Reducing from 10 to 8% RDP decreased dry matter intake (DMI; 0.9 kg/d) at 8% RUP, but increased DMI (2.6 kg/d) at 6% RUP in primiparous cows. Reducing from 10 to 8% RDP decreased milk yield (10%) at 8% RUP, but increased yield (14%) at 6% RUP. Treatments did not affect yield of energy-corrected milk. For multiparous cows, treatments did not affect DMI. Reducing from 10 to 8% RDP decreased yield of energy-corrected milk (3.4%) at 8% RUP, but increased yield (8.8%) at 6% RUP. Reducing from 10 to 8% RDP and 8 to 6% RUP both increased N-use efficiency for primiparous and multiparous cows. The NRC

  2. Mitochondrial Respiratory Defect Causes Dysfunctional Lactate Turnover via AMP-activated Protein Kinase Activation in Human-induced Pluripotent Stem Cell-derived Hepatocytes*

    PubMed Central

    Im, Ilkyun; Jang, Mi-jin; Park, Seung Ju; Lee, Sang-Hee; Choi, Jin-Ho; Yoo, Han-Wook; Kim, Seyun; Han, Yong-Mahn

    2015-01-01

    A defective mitochondrial respiratory chain complex (DMRC) causes various metabolic disorders in humans. However, the pathophysiology of DMRC in the liver remains unclear. To understand DMRC pathophysiology in vitro, DMRC-induced pluripotent stem cells were generated from dermal fibroblasts of a DMRC patient who had a homoplasmic mutation (m.3398T→C) in the mitochondrion-encoded NADH dehydrogenase 1 (MTND1) gene and that differentiated into hepatocytes (DMRC hepatocytes) in vitro. DMRC hepatocytes showed abnormalities in mitochondrial characteristics, the NAD+/NADH ratio, the glycogen storage level, the lactate turnover rate, and AMPK activity. Intriguingly, low glycogen storage and transcription of lactate turnover-related genes in DMRC hepatocytes were recovered by inhibition of AMPK activity. Thus, AMPK activation led to metabolic changes in terms of glycogen storage and lactate turnover in DMRC hepatocytes. These data demonstrate for the first time that energy depletion may lead to lactic acidosis in the DMRC patient by reduction of lactate uptake via AMPK in liver. PMID:26491018

  3. [Value of combined measurement of C-reactive protein, erythrocyte sedimentation rate, lactate dehydrogenase and serum ferritin in etiological diagnosis of fever of unknown origin in children].

    PubMed

    Xie, Ting; Pan, Jia-Hua; Zhang, Xue

    2015-09-01

    To study the clinical value of combined measurement of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), lactate dehydrogenase (LDH), and serum ferritin (SF) in the etiological diagnosis of fever of unknown origin (FUO) in children. The clinical data of 154 hospitalized children who had FUO for at least two weeks were retrospectively analyzed, and they were classified into infection (n=54), rheumatism (n=67), and tumor (n=33) groups according to the diagnosis at discharge. The levels of CRP, ESR, LDH, and SF were compared between the three groups, and the diagnostic values of the four indices alone or together were analyzed using the receiver operating characteristic (ROC) curve. Serum CRP and ESR levels were elevated in all the three groups, and increased most significantly in the rheumatism group. Serum LDH level was increased most significantly in the tumor group. SF level was significantly increased in the rheumatism and tumor groups. The area under the ROC curve (AUC) of LDH for diagnosing rheumatism and the AUC of ESR and CRP for diagnosing tumors were lower than 0.7 (P>0.05). The AUC of CRP for diagnosing infection and rheumatism was 0.861 and 0.782, respectively. The AUC of ESR for diagnosing infection and rheumatism was 0.770 and 0.743, respectively. LDH had relatively low AUC, sensitivity, specificity, and Youden's index in diagnosing infection and tumors. SF had the highest AUC, sensitivity, and Youden's index in diagnosing infection, but had the lowest specificity. SF had relatively high AUC, sensitivity, specificity, and Youden's index in diagnosing rheumatism, but had relatively low AUC in diagnosing tumor. The four indices had higher AUC, sensitivity, and specificity in diagnosing rheumatism and tumors when measured together than when measured alone. In the etiological diagnosis of FUO in children, CRP, ESR, LDH, and SF have certain clinical significance in the preliminary diagnosis of rheumatic diseases, but have limited value in the

  4. Short communication: Supplementing lysine and methionine in a lactation diet containing a high concentration of wet corn gluten feed did not alter milk protein yield.

    PubMed

    Mullins, C R; Weber, D; Block, E; Smith, J F; Brouk, M J; Bradford, B J

    2013-08-01

    Primiparous (n=33) and multiparous (n=63) lactating Holstein cows (186±51 d in milk) were used to evaluate the effects of supplementing metabolizable amino acids using lysine in a matrix of Ca salts of fatty acids (Megamine-L, Arm & Hammer Animal Nutrition, Princeton, NJ) and the isopropyl ester of 2-hydroxy-4-(methylthio) butanoic acid (MetaSmart, Adisseo Inc., Antony, France) in diets containing >26% wet corn gluten feed (dry matter basis). Cows were blocked by production level, parity, and pregnancy status, then randomly assigned to 1 of 8 pens and allowed a 7-d adaption period before receiving dietary treatments for 28 d. Pens were assigned randomly to either of 2 diets formulated to differ by metabolizable amino acid supply. Dry matter intake and production were monitored daily and milk components analyzed 3d/wk. Data were analyzed using mixed models with repeated measures. The original design of the study consisted of a control diet predicted to be deficient in lysine and methionine; however, after ingredient nutrients were analyzed and modeled with animal requirements at dry matter intake [26.6±0.35 kg/d (mean ± SEM)] and milk production levels achieved during the study (40.1±0.46 kg/d), only marginal deficiencies were predicted for the control (-8.1g/d for lysine; -1g/d for methionine) according to the National Research Council method, whereas the Cornell Net Carbohydrate and Protein System 5.0 and 6.1 models indicated positive balances for these amino acids (25.9 and 21.8 g/d for lysine, 14.7 and 18.9 g/d for methionine, respectively). Supplementing 30 g/d of metabolizable lysine in a Ca soap matrix and 2.4 g/d of metabolizable methionine as 2-hydroxy-4-(methylthio) butanoic acid led to positive predicted lysine and methionine balances by all 3 models, and predicted metabolizable lysine-to-methionine ratios ranging from 2.9 to 3.1. No treatment effects were observed for dry matter intake, milk yield, milk component concentrations or yields, or energy

  5. Metabolic regulation by lactate.

    PubMed

    Sola-Penna, Mauro

    2008-09-01

    For more than a century, the metabolic role of lactate has intrigued physiologists and biochemists. Yet, for the first half of the last century lactate had been designated as a waste product, and assigned no additional significance besides its controversial role in muscle fatigue. The decline of the lactate hypothesis for the onset of muscle fatigue and the defining of some modulatory properties attributed to lactate have increased the interest on this molecule. The present critical review aimed at evaluating some recent publications concerned with unveiling the regulatory actions of lactate in cellular function. Lactate has been described to modulate enzymes catalytic properties to affect hormonal release and responsiveness, and to control body homeostasis. Moreover, these properties are directly related to the genesis and the sustainability of pathological conditions, such as diabetes and cancer. In the end, we concluded that lactate should not be regarded as simply an anaerobic metabolite, but should be considered as a regulatory molecule that modulates the integration of metabolism.

  6. Structural Basis for Parathyroid Hormone-related Protein Binding to the Parathyroid Hormone Receptor and Design of Conformation-selective Peptides

    SciTech Connect

    Pioszak, Augen A.; Parker, Naomi R.; Gardella, Thomas J.; Xu, H. Eric

    2009-12-01

    Parathyroid hormone (PTH) and PTH-related protein (PTHrP) are two related peptides that control calcium/phosphate homeostasis and bone development, respectively, through activation of the PTH/PTHrP receptor (PTH1R), a class B G protein-coupled receptor. Both peptides hold clinical interest for their capacities to stimulate bone formation. PTH and PTHrP display different selectivity for two distinct PTH1R conformations, but how their binding to the receptor differs is unclear. The high resolution crystal structure of PTHrP bound to the extracellular domain (ECD) of PTH1R reveals that PTHrP binds as an amphipathic {alpha}-helix to the same hydrophobic groove in the ECD as occupied by PTH, but in contrast to a straight, continuous PTH helix, the PTHrP helix is gently curved and C-terminally 'unwound.' The receptor accommodates the altered binding modes by shifting the side chain conformations of two residues within the binding groove: Leu-41 and Ile-115, the former acting as a rotamer toggle switch to accommodate PTH/PTHrP sequence divergence, and the latter adapting to the PTHrP curvature. Binding studies performed with PTH/PTHrP hybrid ligands having reciprocal exchanges of residues involved in different contacts confirmed functional consequences for the altered interactions and enabled the design of altered PTH and PTHrP peptides that adopt the ECD-binding mode of the opposite peptide. Hybrid peptides that bound the ECD poorly were selective for the G protein-coupled PTH1R conformation. These results establish a molecular model for better understanding of how two biologically distinct ligands can act through a single receptor and provide a template for designing better PTH/PTHrP therapeutics.

  7. Role of Parathyroid Hormone-Related Protein Signaling in Chronic Pancreatitis

    PubMed Central

    Falzon, Miriam; Bhatia, Vandanajay

    2015-01-01

    Chronic pancreatitis (CP), a progressive inflammatory disease where acini are destroyed and replaced by fibrous tissue, increases the risk for pancreatic cancer. Risk factors include alcohol, smoking, and obesity. The effects of these risk factors are exacerbated in patients with mutations in genes that predispose to CP. The different environmental and genetic factors produce the same clinical phenotype; once CP develops, disease course is the same regardless of etiology. Critical questions still need to be answered to understand what modifies predisposition to develop CP in persons exposed to risk factors. We postulate that risk factors modulate endogenous pathways, with parathyroid hormone-related protein (PTHrP) signaling being one such pathway. In support, PTHrP levels are elevated in mice treated with alcohol, and in mouse models of cerulein- and pancreatic duct ligation-induced CP. Disrupting the Pthrp gene in acinar cells exerts protective effects (decreased edema, histological damage, amylase and cytokine release, and fibrosis) in these CP models. PTHrP levels are elevated in human CP. Currently, CP care lacks specific pharmacological interventions. Targeting PTHrP signaling may present a novel therapeutic strategy that inhibits pancreatic inflammation and fibrosis, especially since the risk of developing pancreatic cancer is strongly associated with duration of chronic inflammation. PMID:26095761

  8. Cell-type-specific transactivation of the parathyroid hormone-related protein gene promoter by the human T-cell leukemia virus type I (HTLV-I) tax and HTLV-II tax proteins.

    PubMed

    Ejima, E; Rosenblatt, J D; Massari, M; Quan, E; Stephens, D; Rosen, C A; Prager, D

    1993-02-15

    The human T-cell leukemia virus type I (HTLV-I) and HTLV-II Tax proteins are potent transactivators of viral and cellular gene expression. Using deletion mutants, the downstream parathyroid hormone-related protein (PTHrP) promoter is shown to be responsive to both HTLV-I and HTLV-II Tax as well as the AP1/c-jun proto-oncogene. Transactivation of PTHrP by Tax was seen in T cells but not in B-cell lines or fibroblasts. A carboxy terminal Tax deletion mutant was deficient in transactivation of both the PTHrP and IL2R alpha promoters but not the HTLV-I long terminal repeat (LTR). Exogenous provision of NFkB rescued IL2R alpha expression but not the PTHrP promoter. Thus, HTLV-I Tax, HTLV-II Tax, and c-jun transactivate PTHrP and may contribute to the pathogenesis of hypercalcemia in adult T-cell leukemia.

  9. A comparative proteomic analysis of Bacillus coagulans in response to lactate stress during the production of L-lactic acid.

    PubMed

    Wang, Xiuwen; Qin, Jiayang; Wang, Landong; Xu, Ping

    2014-12-01

    The growth rate and maximum biomass of Bacillus coagulans 2-6 were inhibited by lactate; inhibition by sodium lactate was stronger than by calcium lactate. The differences of protein expressions by B. coagulans 2-6 under the lactate stress were determined using two-dimensional electrophoresis coupled with mass spectrometric identification. Under the non-stress condition, calcium lactate stress and sodium lactate stress, the number of detected protein spots was 1,571 ± 117, 1,281 ± 231 and 904 ± 127, respectively. Four proteins with high expression under lactate stress were identified: lactate dehydrogenase, cysteine synthase A, aldo/keto reductase and ribosomal protein L7/L12. These proteins are thus potential targets for the reconstruction of B. coagulans to promote its resistance to lactate stress.

  10. Transcriptional regulation of parathyroid hormone-related protein promoter P3 by ETS-1 in adult T-cell leukemia/lymphoma.

    PubMed

    Richard, V; Nadella, M V P; Green, P L; Lairmore, M D; Feuer, G; Foley, J G; Rosol, T J

    2005-07-01

    Parathyroid hormone-related protein (PTHrP) plays a primary role in the development of humoral hypercalcemia of malignancy seen in the majority of adult T-cell leukemia/lymphoma (ATLL) patients with human T-cell lymphotropic virus type-1 (HTLV-1) infection. HTLV-1 Tax has been shown to complex with ETS-1 and SP1 to transactivate the PTHrP P3 promoter. Previously, we established a SCID/bg mouse model of human ATL with RV-ATL cells and showed that PTHrP expression was independent of Tax. In this study, we report an inverse correlation of PTHrP with tax/rex mRNA in multiple HTLV-1-positive cell lines and RV-ATL cells. Stimulation of Jurkat T cells with PMA/ionomycin upregulated the PTHrP P3 promoter by a previously characterized Ets binding site and also induced protein/DNA complex formation identical to that observed in RV-ATL cells. Further, we provide evidence that cotransfection with Ets-1 and constitutively active Mek-1 in HTLV-1-negative transformed T cells with stimulation by PMA/ionomycin not only resulted in a robust induction of PTHrP P3 but also formed a complex with ETS-1/P3 EBS similar to that in ATLL cells. Our data demonstrate that transcriptional regulation of PTHrP in ATLL cells can be controlled by T-cell receptor signaling and the ETS and MAPK ERK pathway in a Tax-independent manner.

  11. Sex differences in transgenerational alterations of growth and metabolism in progeny (F2) of female offspring (F1) of rats fed a low protein diet during pregnancy and lactation.

    PubMed

    Zambrano, E; Martínez-Samayoa, P M; Bautista, C J; Deás, M; Guillén, L; Rodríguez-González, G L; Guzmán, C; Larrea, F; Nathanielsz, P W

    2005-07-01

    Compelling epidemiological and experimental evidence indicates that a suboptimal environment during fetal and neonatal development in both humans and animals may programme offspring susceptibility to later development of several chronic diseases including obesity and diabetes in which altered carbohydrate metabolism plays a central role. One of the most interesting and significant features of developmental programming is the evidence from several studies that the adverse consequences of altered intrauterine environments can be passed transgenerationally from mother (F0) to daughter (F1) to second generation offspring (F2). We determined whether when F0 female rats are exposed to protein restriction during pregnancy and/or lactation their F1 female pups deliver F2 offspring with in vivo evidence of altered glucose and insulin metabolism. We fed F0 virgin Wistar rats a normal control 20% casein diet (C) or a protein restricted isocaloric diet (R) containing 10% casein during pregnancy. F1 female R pups weighed less than C at birth. After delivery, mothers received C or R diet during lactation to provide four F1 offspring groups CC (first letter pregnancy diet and second lactation diet), RR, CR and RC. All F1 female offspring were fed ad libitum with C diet after weaning and during their first pregnancy and lactation. As they grew female offspring (F1) of RR and CR mothers exhibited low body weight and food intake with increased sensitivity to insulin during a glucose tolerance test at 110 days of postnatal life. Male F2 CR offspring showed evidence of insulin resistance. In contrast RC F2 females showed evidence of insulin resistance. Sex differences were also observed in F2 offspring in resting glucose and insulin and insulin: glucose ratios. These sex differences also showed differences specific to stage of development time window. We conclude that maternal protein restriction adversely affects glucose and insulin metabolism of male and female F2 offspring in a

  12. Crystallization of the receptor-binding domain of parathyroid hormone-related protein in complex with a neutralizing monoclonal antibody Fab fragment

    SciTech Connect

    McKinstry, William J.; Polekhina, Galina; Diefenbach-Jagger, Hannelore; Sato, Koh; Onuma, Etsuro; Gillespie, Matthew T.; Martin, Thomas J.; Parker, Michael W.

    2009-04-01

    Parathyroid hormone-related protein (PTHrP) plays an important role in regulating embryonic skeletal development and is abnormally regulated in the pathogenesis of skeletal complications observed with many cancers and osteoporosis. It exerts its action through binding to a G-protein-coupled seven-transmembrane cell-surface receptor (GPCR). Structurally, GPCRs are very difficult to study by X-ray crystallography. In this study, a monoclonal antibody Fab fragment which recognizes the same region of PTHrP as its receptor, PTH1R, was used to aid in the crystallization of PTHrP. The resultant protein complex was crystallized using the hanging-drop vapour-diffusion method with polyethylene glycol as a precipitant. The crystals belonged to the orthorhombic space group P2{sub 1}2{sub 1}2, with unit-cell parameters a = 72.6, b = 96.3, c = 88.5 {angstrom}, and diffracted to 2.0 {angstrom} resolution using synchrotron radiation. The crystal structure will shed light on the nature of the key residues of PTHrP that interact with the antibody and will provide insights into how the antibody is able to discriminate between PTHrP and the related molecule parathyroid homone.

  13. Crystallization of the receptor-binding domain of parathyroid hormone-related protein in complex with a neutralizing monoclonal antibody Fab fragment.

    PubMed

    McKinstry, William J; Polekhina, Galina; Diefenbach-Jagger, Hannelore; Sato, Koh; Onuma, Etsuro; Gillespie, Matthew T; Martin, Thomas J; Parker, Michael W

    2009-04-01

    Parathyroid hormone-related protein (PTHrP) plays an important role in regulating embryonic skeletal development and is abnormally regulated in the pathogenesis of skeletal complications observed with many cancers and osteoporosis. It exerts its action through binding to a G-protein-coupled seven-transmembrane cell-surface receptor (GPCR). Structurally, GPCRs are very difficult to study by X-ray crystallography. In this study, a monoclonal antibody Fab fragment which recognizes the same region of PTHrP as its receptor, PTH1R, was used to aid in the crystallization of PTHrP. The resultant protein complex was crystallized using the hanging-drop vapour-diffusion method with polyethylene glycol as a precipitant. The crystals belonged to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 72.6, b = 96.3, c = 88.5 A, and diffracted to 2.0 A resolution using synchrotron radiation. The crystal structure will shed light on the nature of the key residues of PTHrP that interact with the antibody and will provide insights into how the antibody is able to discriminate between PTHrP and the related molecule parathyroid homone.

  14. Developmental programming of aging of isolated pancreatic islet glucose-stimulated insulin secretion in female offspring of mothers fed low-protein diets in pregnancy and/or lactation.

    PubMed

    Morimoto, S; Sosa, T C; Calzada, L; Reyes-Castro, L A; Díaz-Díaz, E; Morales, A; Nathanielsz, P W; Zambrano, E

    2012-12-01

    Diabetes predisposition is determined by pancreatic islet insulin secretion and insulin resistance. We studied female rat offspring exposed to low-protein maternal diet (50% control protein diet) in pregnancy and/or lactation at postnatal days 36, 110 and 450. Rats were fed either control 20% casein diet (C) or restricted diet (R - 10% casein) during pregnancy. After delivery, mothers received either C or R diet until weaning to provide four offspring groups: CC, RR, CR and RC (first letter denoting maternal pregnancy diet and the second lactation diet). Serum glucose, insulin and homeostatic model assessment (HOMA) were measured. Pancreatic islets were isolated and in vitro insulin secretion quantified in low glucose (5 mM) and high glucose (11 mM). Serum glucose, insulin and HOMA were similar in all groups at 36 and 110 postnatal days. HOMA was only higher in RR at 450 postnatal days. Only CC demonstrated differences in glucose sensitivity of β-cells to high and low doses at the three ages studied. At 36 days, RR, CR and RC and at 450 days RR and RC groups did not show glucose-stimulated insulin secretion differences between low and high glucose. Aging-associated glucose-stimulated insulin secretion loss was affected by maternal dietary history, indicating that developmental programming must be considered a major factor in aging-related development of predisposition to later-life dysfunctional insulin metabolism. Female offspring islets' insulin secretion was higher than previously reported in males.

  15. Lactation in Islam.

    PubMed

    Hefnawi, F I

    1982-01-01

    Preservation and promotion of breastfeeding in Islamic countries could be increased by stressing the religious importance of this practice as prescribed in Islamic religious teachings. The child's right to be breastfed is affirmed by the Quaran, the source of Islamic law and morality. Quranic verse 2:233 recommends a 2 year period of lactation. According to Islam a nursing mother is entitled to receive compensation from the father for nursing the child. The father, though, has the option to engage a paid or unpaid wet-nurse for the child, in which case the mother looses her right to be paid for nursing even is she volunteered to breastfeed. The mother's right to nurse a child without compensation is prior to a father's right to engage a wet-nurse. In another Islamic source the moral importance of breastfeeding is stressed. The mother receives the reward of a good deed for every single drop she gives her child. Islamic precepts on lactation influenced Arabian medicine. Avicenna's view that children should be breastfed for 2 years was approvingly quoted by European physicians in the 17th century. Major Arabian medical texts contain chapters on lactation, on tests for quality of breast milk, and on diets and drugs for improving lactation. Research at Al-Azhar University is directed toward finding a contraceptive that will not inhabit lactation and will not affect the quality of breast milk.

  16. Lactate: Friend or Foe.

    PubMed

    Hall, Mederic M; Rajasekaran, Sathish; Thomsen, Timothy W; Peterson, Andrew R

    2016-03-01

    Lactic acid has played an important role in the traditional theory of muscle fatigue and limitation of endurance exercise performance. It has been called a waste product of anaerobic metabolism and has been believed to be responsible for the uncomfortable "burn" of intense exercise and directly responsible for the metabolic acidosis of exercise, leading to decreased muscle contractility and ultimately cessation of exercise. Although this premise has been commonly taught, it is not supported by the scientific literature and has led to a great deal of confusion among the sports medicine and exercise science communities. This review will provide the sports medicine clinician with an understanding of contemporary lactate theories, including lactate's role in energy production, its contributions to metabolic acidosis, and its function as an energy substrate for a variety of tissues. Lactate threshold concepts will also be discussed, including a practical approach to understanding prediction of performance and monitoring of training progress based on these parameters.

  17. Maternal protein restriction during lactation modulated the expression and activity of rat offspring hepatic CYP1A1, CYP1A2, CYP2B1, CYP2B2, and CYP2E1 during development.

    PubMed

    Da Costa, N Meireles; Visoni, S B C; Dos Santos, I L; Barja-Fidalgo, T C; Ribeiro-Pinto, L F

    2016-01-01

    Early nutrition plays a long-term role in the predisposition to chronic diseases and influences the metabolism of several drugs. This may happen through cytochromes P450 (CYPs) regulation, which are the main enzymes responsible for the metabolism of xenobiotics. Here, we analyzed the effects of maternal protein restriction (MPR) on the expression and activity of hepatic offspring's CYPs during 90 days after birth, using Wistar rats as a mammal model. Hepatic CYP1A1, CYP1A2, CYP2B1, CYP2B2 and CYP2E1 mRNA and protein expression, and associated catalytic activities (ECOD, EROD, MROD, BROD, PROD and PNPH) were evaluated in 15-, 30-, 60-, and 90-day-old offspring from dams fed with either a 0% protein (MPR groups) or a standard diet (C groups) during the 10 first days of lactation. Results showed that most CYP genes were induced in 60- and 90-day-old MPR offspring. The inductions detected in MPR60 and MPR90 were of 5.0- and 2.0-fold (CYP1A2), 3.7- and 2.0-fold (CYP2B2) and 9.8- and 5.8- fold (CYP2E1), respectively, and a 3.8-fold increase of CYP2B1 in MPR90. No major alterations were detected in CYP protein expression. The most relevant CYP catalytic activities' alterations were observed in EROD, BROD and PNPH. Nevertheless, they did not follow the same pattern observed for mRNA expression, except for an induction of EROD in MPR90 (3.5-fold) and of PNPH in MPR60 (2.2-fold). Together, these results suggest that MPR during lactation was capable of altering the expression and activity of the hepatic CYP enzymes evaluated in the offspring along development.

  18. Maternal protein restriction during lactation modulated the expression and activity of rat offspring hepatic CYP1A1, CYP1A2, CYP2B1, CYP2B2, and CYP2E1 during development

    PubMed Central

    Da Costa, N. Meireles; Visoni, S.B.C.; Dos Santos, I.L.; Barja-Fidalgo, T.C.; Ribeiro-Pinto, L.F.

    2016-01-01

    Early nutrition plays a long-term role in the predisposition to chronic diseases and influences the metabolism of several drugs. This may happen through cytochromes P450 (CYPs) regulation, which are the main enzymes responsible for the metabolism of xenobiotics. Here, we analyzed the effects of maternal protein restriction (MPR) on the expression and activity of hepatic offspring’s CYPs during 90 days after birth, using Wistar rats as a mammal model. Hepatic CYP1A1, CYP1A2, CYP2B1, CYP2B2 and CYP2E1 mRNA and protein expression, and associated catalytic activities (ECOD, EROD, MROD, BROD, PROD and PNPH) were evaluated in 15-, 30-, 60-, and 90-day-old offspring from dams fed with either a 0% protein (MPR groups) or a standard diet (C groups) during the 10 first days of lactation. Results showed that most CYP genes were induced in 60- and 90-day-old MPR offspring. The inductions detected in MPR60 and MPR90 were of 5.0- and 2.0-fold (CYP1A2), 3.7- and 2.0-fold (CYP2B2) and 9.8- and 5.8– fold (CYP2E1), respectively, and a 3.8-fold increase of CYP2B1 in MPR90. No major alterations were detected in CYP protein expression. The most relevant CYP catalytic activities’ alterations were observed in EROD, BROD and PNPH. Nevertheless, they did not follow the same pattern observed for mRNA expression, except for an induction of EROD in MPR90 (3.5-fold) and of PNPH in MPR60 (2.2-fold). Together, these results suggest that MPR during lactation was capable of altering the expression and activity of the hepatic CYP enzymes evaluated in the offspring along development. PMID:27828666

  19. Contraception in lactating women.

    PubMed

    Díaz, S; Croxatto, H B

    1993-12-01

    Lactating women need contraception after first menses, supplementation or 6 months postpartum, or before, according to personal or programmatic reasons. Non-hormonal methods have no influence on lactation and are the first choice. Intrauterine devices (IUDs) inserted during amenorrhea are safe and show good continuation rates. Progestin-only methods do not affect breast-feeding and should be used after 6 weeks to prevent transference of orally active steroids to the newborn. Progesterone rings and Nestorone implants are effective new methods, that use orally inactive steroids.

  20. Cellular mechanism through which parathyroid hormone-related protein induces proliferation in arterial smooth muscle cells: definition of an arterial smooth muscle PTHrP/p27kip1 pathway.

    PubMed

    Fiaschi-Taesch, Nathalie; Sicari, Brian M; Ubriani, Kiran; Bigatel, Todd; Takane, Karen K; Cozar-Castellano, Irene; Bisello, Alessandro; Law, Brian; Stewart, Andrew F

    2006-10-27

    Parathyroid hormone-related protein (PTHrP) is present in vascular smooth muscle (VSM), is markedly upregulated in response to arterial injury, is essential for normal VSM proliferation, and also markedly accentuates neointima formation following rat carotid angioplasty. PTHrP contains a nuclear localization signal (NLS) through which it enters the nucleus and leads to marked increases in retinoblastoma protein (pRb) phosphorylation and cell cycle progression. Our goal was to define key cell cycle molecules upstream of pRb that mediate cell cycle acceleration induced by PTHrP. The cyclin D/cdk-4,-6 system and its upstream regulators, the inhibitory kinases (INKs), are not appreciably influenced by PTHrP. In striking contrast, cyclin E/cdk-2 kinase activity is markedly increased by PTHrP, and this is a result of a specific, marked, PTHrP-induced proteasomal degradation of p27(kip1). Adenoviral restoration of p27(kip1) fully reverses PTHrP-induced cell cycle progression, indicating that PTHrP mediates its cell cycle acceleration in VSM via p27(kip1). In confirmation, adenoviral delivery of PTHrP to murine primary vascular smooth muscle cells (VSMCs) significantly decreases p27(kip1) expression and accelerates cell cycle progression. p27(kip1) is well known to be a central cell cycle regulatory molecule involved in both normal and pathological VSM proliferation and is a target of widely used drug-eluting stents. The current observations define a novel "PTHrP/p27(kip1) pathway" in the arterial wall and suggest that this pathway is important in normal arterial biology and a potential target for therapeutic manipulation of the arterial response to injury.

  1. Glycoprotein expression in human milk during lactation.

    PubMed

    Froehlich, John W; Dodds, Eric D; Barboza, Mariana; McJimpsey, Erica L; Seipert, Richard R; Francis, Jimi; An, Hyun Joo; Freeman, Samara; German, J Bruce; Lebrilla, Carlito B

    2010-05-26

    While milk proteins have been studied for decades, strikingly little effort has been applied to determining how the post-translational modifications (PTMs) of these proteins may change during the course of lactation. PTMs, particularly glycosylation, can greatly influence protein structure, function, and stability and can particularly influence the gut where their degradation products are potentially bioactive. In this work, previously undiscovered temporal variations in both expression and glycosylation of the glycoproteome of human milk are observed. Lactoferrin, one of the most abundant glycoproteins in human milk, is shown to be dynamically glycosylated during the first 10 days of lactation. Variations in expression or glycosylation levels are also demonstrated for several other abundant whey proteins, including tenascin, bile salt-stimulated lipase, xanthine dehydrogenase, and mannose receptor.

  2. Productive life including all lactations and longer lactations with diminishing credits.

    PubMed

    VanRaden, P M; Dematawewa, C M B; Pearson, R E; Tooker, M E

    2006-08-01

    Alternative measures of productive life (PL) were compared, and life expectancy factors were updated to replace estimates from 1993. Alternatives were proposed with extra credits for lactations longer than 10 mo and beyond 84 mo of age and for each calving so that an extremely long lactation would not receive more credits than multiple shorter lactations with dry periods between. Maximum credits per lactation of 10 mo (original PL), 12 mo, and unlimited were compared. The unlimited credits option either included or excluded a calf value equal to 2 mo of production and had credits given for all days either uniformly or based on lactation curves (diminishing credits). Standard lactation curves (first, second, and greater lactations) were estimated based on the test-day yields of Holstein cows remaining in lactation from a set of 903,579 lactation records. For the diminishing credits alternative, credit for a given day of a parity was derived using the predicted yield of the day proportional to the average daily yield of the first 305 d of second parity. Daily yields were deviations from a baseline of 13.62 kg. Heritabilities and genetic correlations were estimated by multitrait REML for alternative measures of PL, for longevity censored at various ages, and for yield traits and SCS in first parity. Data for REML analysis included records from 1,098,329 Holsteins born from 1994 through 1997 from 5,109 sires, and a relationship matrix among sires was included in the model. Lactations beyond 84 mo added little information. Heritability of PL was 0.073 with 10 mo, 0.069 with 12 mo, 0.068 and 0.067 with unlimited (uniform) lactation credits (with and without calf credits, respectively), and 0.070 with unlimited diminishing credits. Corresponding correlations among predicted transmitting abilities for PL and protein yield were 0.07, 0.06, 0.12, 0.23, and 0.09, all much lower than the 0.46 estimated in 1993. Heritability of PL with diminishing credits improved from 0.017 to

  3. At the crossroads: EGFR and PTHrP signaling in cancer-mediated diseases of bone

    PubMed Central

    Nickerson, Nicole; Riese, David J.; Hollenhorst, Peter C.; Lorch, Gwendolen; Foley, Anne M.

    2014-01-01

    The epidermal growth factor receptor is a well-established cancer therapeutic target due to its stimulation of proliferation, motility, and resistance to apoptosis. Recently, additional roles for the receptor have been identified in growth of metastases. Similar to development, metastatic spread requires signaling interactions between epithelial-derived tumor cells and mesenchymal derivatives of the microenvironment. This necessitates reactivation of developmental signaling molecules, including the hypercalcemia factor parathyroid hormone-related protein. This review covers the variations of epidermal growth factor receptor signaling in cancers that produce bone metastases, regulation of parathyroid hormone-related protein, and evidence that the two molecules drive cancer-mediated diseases of bone. PMID:22684584

  4. 21 CFR 184.1311 - Ferrous lactate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... with iron filings, reaction of ferrous chloride with sodium lactate, or reaction of ferrous sulfate... Substances Affirmed as GRAS § 184.1311 Ferrous lactate. (a) Ferrous lactate (iron (II) lactate, C6H10FeO6...

  5. Interspecies comparison of renal cortical receptors for parathyroid hormone and parathyroid hormone-related protein

    SciTech Connect

    Orloff, J.J.; Goumas, D.; Wu, T.L.; Stewart, A.F. )

    1991-03-01

    Parathyroid hormone (PTH) and PTH-related proteins (PTHrP) interact with a common receptor in rat bone cells and in canine renal membranes with similar affinity, but PTHrP are substantially less potent than PTH in stimulating adenylate cyclase in canine renal membranes; in contrast, PTH and PTHrP are equipotent in stimulating adenylate cyclase in rat bone cells. This discrepancy has been largely viewed as reflecting differences in the relative efficiency of signal transduction of PTHrP between bone and kidney assay systems. To test the alternative (but not mutually exclusive) hypothesis that these differences could reflect interspecies differences in PTH receptors, we have characterized the bioactivity of amino-terminal PTHrP and PTH in rat and human renal cortical membranes (RCM) and compared them to results we previously reported in canine RCM. The stability of PTH and PTHrP peptides under binding and adenylate cyclase assay conditions was greater than 80% for each species. Competitive inhibition of ({sup 125}I)(Tyr36)hPTHrP-(1-36)NH{sub 2} binding to rat RCM by bPTH-(1-34) and (Tyr36)hPTHrP-(1-36)NH{sub 2} yielded nearly identical binding dissociation constants (3.7 and 3.6 nM, respectively), and binding to human RCM demonstrated slightly greater potency for PTHrP (0.5 nM) than for PTH (0.9 nM). Similarly, adenylate cyclase stimulating activity was equivalent for the two peptides in rat RCM, but PTHrP was twofold more potent than PTH in human RCM. Covalent photoaffinity labeling of protease-protected rat RCM yielded an apparent 80 kD receptor protein, and cross-linking of human RCM labeled an 85 kD receptor, indistinguishable in size from the canine renal PTH receptor. We conclude that rat, canine, and human renal cortical PTH receptors exhibit species specificity.

  6. Parathyroid hormone related-protein promotes epithelial-to-mesenchymal transition in prostate cancer.

    PubMed

    Ongkeko, Weg M; Burton, Doug; Kiang, Alan; Abhold, Eric; Kuo, Selena Z; Rahimy, Elham; Yang, Meng; Hoffman, Robert M; Wang-Rodriguez, Jessica; Deftos, Leonard J

    2014-01-01

    Parathyroid hormone-related protein (PTHrP) possesses a variety of physiological and developmental functions and is also known to facilitate the progression of many common cancers, notably their skeletal invasion, primarily by increasing bone resorption. The purpose of this study was to determine whether PTHrP could promote epithelial-to-mesenchymal transition (EMT), a process implicated in cancer stem cells that is critically involved in cancer invasion and metastasis. EMT was observed in DU 145 prostate cancer cells stably overexpressing either the 1-141 or 1-173 isoform of PTHrP, where there was upregulation of Snail and vimentin and downregulation of E-cadherin relative to parental DU 145. By contrast, the opposite effect was observed in PC-3 prostate cancer cells where high levels of PTHrP were knocked-down via lentiviral siRNA transduction. Increased tumor progression was observed in PTHrP-overexpressing DU 145 cells while decreased progression was observed in PTHrP-knockdown PC-3 cells. PTHrP-overexpressing DU 145 formed larger tumors when implanted orthoptopically into nude mice and in one case resulted in spinal metastasis, an effect not observed among mice injected with parental DU 145 cells. PTHrP-overexpressing DU 145 cells also caused significant bone destruction when injected into the tibiae of nude mice, while parental DU 145 cells caused little to no destruction of bone. Together, these results suggest that PTHrP may work through EMT to promote an aggressive and metastatic phenotype in prostate cancer, a pathway of importance in cancer stem cells. Thus, continued efforts to elucidate the pathways involved in PTHrP-induced EMT as well as to develop ways to specifically target PTHrP signaling may lead to more effective therapies for prostate cancer.

  7. 21 CFR 184.1311 - Ferrous lactate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Ferrous lactate. 184.1311 Section 184.1311 Food... Specific Substances Affirmed as GRAS § 184.1311 Ferrous lactate. (a) Ferrous lactate (iron (II) lactate.... It is prepared by reacting calcium lactate or sodium lactate with ferrous sulfate, direct reaction...

  8. 21 CFR 184.1311 - Ferrous lactate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Ferrous lactate. 184.1311 Section 184.1311 Food... GRAS § 184.1311 Ferrous lactate. (a) Ferrous lactate (iron (II) lactate, C6H10FeO6, CAS Reg. No. 5905... reacting calcium lactate or sodium lactate with ferrous sulfate, direct reaction of lactic acid with...

  9. 21 CFR 184.1311 - Ferrous lactate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Ferrous lactate. 184.1311 Section 184.1311 Food... Specific Substances Affirmed as GRAS § 184.1311 Ferrous lactate. (a) Ferrous lactate (iron (II) lactate.... It is prepared by reacting calcium lactate or sodium lactate with ferrous sulfate, direct reaction...

  10. 21 CFR 184.1311 - Ferrous lactate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Ferrous lactate. 184.1311 Section 184.1311 Food... Specific Substances Affirmed as GRAS § 184.1311 Ferrous lactate. (a) Ferrous lactate (iron (II) lactate.... It is prepared by reacting calcium lactate or sodium lactate with ferrous sulfate, direct reaction...

  11. Single mutation in Shine-Dalgarno-like sequence present in the amino terminal of lactate dehydrogenase of Plasmodium effects the production of an eukaryotic protein expressed in a prokaryotic system.

    PubMed

    Cicek, Mustafa; Mutlu, Ozal; Erdemir, Aysegul; Ozkan, Ebru; Saricay, Yunus; Turgut-Balik, Dilek

    2013-06-01

    One of the most important step in structure-based drug design studies is obtaining the protein in active form after cloning the target gene. In one of our previous study, it was determined that an internal Shine-Dalgarno-like sequence present just before the third methionine at N-terminus of wild type lactate dehydrogenase enzyme of Plasmodium falciparum prevent the translation of full length protein. Inspection of the same region in P. vivax LDH, which was overproduced as an active enzyme, indicated that the codon preference in the same region was slightly different than the codon preference of wild type PfLDH. In this study, 5'-GGAGGC-3' sequence of P. vivax that codes for two glycine residues just before the third methionine was exchanged to 5'-GGAGGA-3', by mimicking P. falciparum LDH, to prove the possible effects of having an internal SD-like sequence when expressing an eukaryotic protein in a prokaryotic system. Exchange was made by site-directed mutagenesis. Results indicated that having two glycine residues with an internal SD-like sequence (GGAGGA) just before the third methionine abolishes the enzyme activity due to the preference of the prokaryotic system used for the expression. This study emphasizes the awareness of use of a prokaryotic system to overproduce an eukaryotic protein.

  12. Maternal consumption of high-prebiotic fibre or -protein diets during pregnancy and lactation differentially influences satiety hormones and expression of genes involved in glucose and lipid metabolism in offspring in rats.

    PubMed

    Maurer, Alannah D; Reimer, Raylene A

    2011-02-01

    Risk of developing the metabolic syndrome may be influenced by nutritional environment early in life. We examined the effects of high-fibre (HF) and high-protein (HP) diets consumed during pregnancy and lactation on satiety hormones and expression of genes involved in glucose and lipid metabolism in offspring. Wistar dams were fed a control (C), HF or HP diets during pregnancy and lactation. At parturition, litters were culled to ten pups. At 21 d, all pups were weaned onto C diet. At 7, 14, 21, 28 and 35 d after birth, blood was analysed for satiety hormones and tissues for mRNA expression in offspring. No differences were observed in litter size or birth weight. At 21 d, offspring of HF dams had greater adjusted intestinal mass and lower liver weight than those of C but not of HP dams. Plasma glucose at 28 d and amylin at 7, 14 and 28 d were lower in HF v. C and HP offspring. Glucagon-like peptide-1 was higher in HP offspring than in HF offspring at 7 d but was higher in HF v. C offspring at 21 d. Offspring of HF dams had higher glucose transporter (GLUT2 and Na+-dependent glucose/galactose transporter) mRNA expression at 21 d v. C and HP offspring. In brown adipose tissue, HF and HP up-regulated uncoupling protein-1 and PPAR-γ coactivator. HP was associated with increased resistin and IL-6 mRNA expression. The present study demonstrates that maternal diet composition differentially regulates circulating satiety hormones and genes involved in glucose transport and energy metabolism in offspring. These early changes could have long-term consequences for obesity risk.

  13. Lactate metabolism in acute uremia.

    PubMed

    Leverve, Xavier; Mustafa, Iqbal; Novak, Ivan; Krouzecky, Ales; Rokyta, Richard; Matejovic, Martin; Ichai, Carole

    2005-01-01

    Lactate is a key metabolite that is produced by every cell and oxidized by most of them, provided that they do contain mitochondria. Its metabolism is connected to energetic homeostasis and the cellular redox state. It is well recognized as an indicator of severe outcome in severely ill patients, however, it is not a detrimental factor per se. Conversely, some recent data tend even to indicate a beneficial effect in several metabolic disorders. Although the liver has long been recognized as a key organ in lactate homeostasis, the kidney also plays a major role as a gluconeogenic organ significantly involved in the glucose-lactate cycle. In acute renal failure, sodium lactate is widely used as a buffer in replacement fluids because the anion (lactate - ) is metabolized and the cation (Na + ) remains, leading to decreased water dissociation and proton concentration. The metabolic disorders related to acute renal failure or associated with it, such as liver failure, may affect lactate metabolism, and therefore they are often regarded as limiting factors for the use of lactate-containing fluids in such patients. By investigating endogenous lactate production in severe septic patients with acute renal failure, we found that an acute exogenous load of lactate did not affect the basal endogenous lactate production and metabolism. This indicates that exogenous lactate is well metabolized even in patients suffering from acute renal failure and severe sepsis with a compromised hemodynamic status.

  14. Treatment with N- and C-Terminal Peptides of Parathyroid Hormone-Related Protein Partly Compensate the Skeletal Abnormalities in IGF-I Deficient Mice

    PubMed Central

    Portal-Núñez, Sergio; Murillo-Cuesta, Silvia; Lozano, Daniel; Cediel, Rafael; Esbrit, Pedro

    2014-01-01

    Insulin-like growth factor-I (IGF-I) deficiency causes growth delay, and IGF-I has been shown to partially mediate bone anabolism by parathyroid hormone (PTH). PTH-related protein (PTHrP) is abundant in bone, and has osteogenic features by poorly defined mechanisms. We here examined the capacity of PTHrP (1–36) and PTHrP (107–111) (osteostatin) to reverse the skeletal alterations associated with IGF-I deficiency. Igf1-null mice and their wild type littermates were treated with each PTHrP peptide (80 µg/Kg/every other day/2 weeks; 2 males and 4 females for each genotype) or saline vehicle (3 males and 3 females for each genotype). We found that treatment with either PTHrP peptide ameliorated trabecular structure in the femur in both genotypes. However, these peptides were ineffective in normalizing the altered cortical structure at this bone site in Igf1-null mice. An aberrant gene expression of factors associated with osteoblast differentiation and function, namely runx2, osteoprotegerin/receptor activator of NF-κB ligand ratio, Wnt3a, cyclin D1, connexin 43, catalase and Gadd45, as well as in osteocyte sclerostin, was found in the long bones of Igf1-null mice. These mice also displayed a lower amount of trabecular osteoblasts and osteoclasts in the tibial metaphysis than those in wild type mice. These alterations in Igf1-null mice were only partially corrected by each PTHrP peptide treatment. The skeletal expression of Igf2, Igf1 receptor and Irs2 was increased in Igf1-null mice, and this compensatory profile was further improved by treatment with each PTHrP peptide related to ERK1/2 and FoxM1 activation. In vitro, PTHrP (1–36) and osteostatin were effective in promoting bone marrow stromal cell mineralization in normal mice but not in IGF-I-deficient mice. Collectively, these findings indicate that PTHrP (1–36) and osteostatin can exert several osteogenic actions even in the absence of IGF-I in the mouse bone. PMID:24503961

  15. Changes in Llama (Lama glama) milk composition during lactation.

    PubMed

    Riek, A; Gerken, M

    2006-09-01

    Milk samples were collected weekly from 10 llamas during the first 27 wk after parturition under controlled stable conditions. Mean values for the concentrations of the major milk components across the lactation period were 4.70% fat, 4.23% protein, 5.93% lactose, 15.61% dry matter, and 22.62 mg/dL of milk urea N. All constituents were affected by the stage of lactation. There was an increase in fat to protein ratio as protein concentration declined and fat concentration increased. Fat, protein, and lactose concentrations changed during the transition from colostrum to milk. In the first month postpartum, fat concentration remained constant, protein decreased, and lactose increased. Starting with wk 5 postpartum, fat and protein increased and lactose decreased until the end of lactation. Among the major constituents fat had the highest variation. The mean gross energy concentration of milk was 3.88 kJ/g and showed a similar course as protein. Fat contributed 48.0%, protein 26.3%, and lactose 25.7% to the gross energy in the milk. Milk urea N values were higher than those found in ruminants and increased with stage of lactation, whereas the pH decreased. The analyzed milk components were not affected by the lactation number of the animal, except milk urea N. Somatic cell counts indicated the absence of mastitis and revealed that the average somatic cell count of uninfected llamas is lower than in animals usually used for milk production. The 2 algebraic models fitted by a nonlinear regression procedure to the data resulted in suitable prediction curves for the constituents (R2 = 0.76 to 0.94). The courses of major milk constituents in llamas during lactation are similar to those in domesticated ruminants, although different in their values. The established curves facilitate the composition of milk replacers at different stages of lactation for nursing llamas whose dams died or are agalactic.

  16. Gastrointestinal transport of calcium and glucose in lactating ewes.

    PubMed

    Klinger, Stefanie; Schröder, Bernd; Gemmer, Anja; Reimers, Julia; Breves, Gerhard; Herrmann, Jens; Wilkens, Mirja R

    2016-06-01

    During lactation, mineral and nutrient requirements increase dramatically, particularly those for Ca and glucose. In contrast to monogastric species, in ruminants, it is rather unclear to which extend this physiological change due to increased demand for milk production is accompanied by functional adaptations of the gastrointestinal tract (GIT). Therefore, we investigated potential modulations of Ca and glucose transport mechanisms in the GIT of lactating and dried-off sheep. Ussing-chamber technique was applied to determine the ruminal and jejunal Ca flux rates. In the jejunum, electrophysiological properties in response to glucose were recorded. Jejunal brush-border membrane vesicles (BBMV) served to characterize glucose uptake via sodium-linked glucose transporter 1 (SGLT1), and RNA and protein expression levels of Ca and glucose transporting systems were determined. Ruminal Ca flux rate data showed a trend for higher absorption in lactating sheep. In the jejunum, small Ca absorption could only be observed in lactating ewes. From the results, it may be assumed that lactating ewes compensate for the Ca loss by increasing bone mobilization rather than by increasing supply through absorption from the GIT Presence of SGLT1 in the jejunum of both groups was shown by RNA and protein identification, but glucose uptake into BBMV could only be detected in lactating sheep. This, however, could not be attributed to electrogenic glucose absorption in lactating sheep under Ussing-chamber conditions, providing evidence that changes in jejunal glucose uptake may include additional factors, that is, posttranslational modifications such as phosphorylation.

  17. PTH-related protein upregulates integrin {alpha}6{beta}4 expression and activates Akt in breast cancer cells

    SciTech Connect

    Shen Xiaoli; Falzon, Miriam . E-mail: mfalzon@utmb.edu

    2006-11-15

    Breast cancer is the most common carcinoma that metastasizes to bone. Tumor-produced parathyroid hormone-related protein (PTHrP), a known stimulator of osteoclastic bone resorption, is a major mediator of the osteolytic process in breast cancer. We have previously shown that PTHrP increases breast cancer cell proliferation, survival, migration, and pro-invasive integrin {alpha}6{beta}4 expression. To determine the role of integrin {alpha}6{beta}4 in these PTHrP-mediated effects, we utilized two strategies to modulate expression of the {alpha}6 and {beta}4 subunits in parental and PTHrP-overexpressing MDA-MB-231 and MCF-7 cells: overexpression of {alpha}6{beta}4 by transfection with constructs encoding the {alpha}6 and {beta}4 subunits, and suppression of endogenous {alpha}6{beta}4 expression by transfection with siRNAs targeting these subunits. We now show that the effects of PTHrP are mediated via upregulation of integrin {alpha}6{beta}4 expression. We also show that integrin {alpha}6{beta}4 expression is modulated at the mRNA level, indicating a transcriptional and/or post-transcriptional mechanism of action for PTHrP. PTHrP expression also increased the levels of phosphorylated Akt, with a consequent increase in the levels of phosphorylated (inactive) glycogen synthase kinase-3 (GSK-3). The role of PTHrP in breast cancer growth and metastasis may thus be mediated via upregulation of integrin {alpha}6{beta}4 expression and Akt activation, with consequent inactivation of GSK-3.

  18. Microbial production of lactate-containing polyesters

    PubMed Central

    Yang, Jung Eun; Choi, So Young; Shin, Jae Ho; Park, Si Jae; Lee, Sang Yup

    2013-01-01

    Due to our increasing concerns on environmental problems and limited fossil resources, biobased production of chemicals and materials through biorefinery has been attracting much attention. Optimization of the metabolic performance of microorganisms, the key biocatalysts for the efficient production of the desired target bioproducts, has been achieved by metabolic engineering. Metabolic engineering allowed more efficient production of polyhydroxyalkanoates, a family of microbial polyesters. More recently, non-natural polyesters containing lactate as a monomer have also been produced by one-step fermentation of engineered bacteria. Systems metabolic engineering integrating traditional metabolic engineering with systems biology, synthetic biology, protein/enzyme engineering through directed evolution and structural design, and evolutionary engineering, enabled microorganisms to efficiently produce natural and non-natural products. Here, we review the strategies for the metabolic engineering of microorganisms for the in vivo biosynthesis of lactate-containing polyesters and for the optimization of whole cell metabolism to efficiently produce lactate-containing polyesters. Also, major problems to be solved to further enhance the production of lactate-containing polyesters are discussed. PMID:23718266

  19. Metabolic Bone Disease in the Context of Metastatic Neuroendocrine Tumor: Differentiation from Skeletal Metastasis, the Molecular PET–CT Imaging Features, and Exploring the Possible Etiopathologies Including Parathyroid Adenoma (MEN1) and Paraneoplastic Humoral Hypercalcemia of Malignancy Due to PTHrP Hypersecretion

    PubMed Central

    Ranade, Rohit; Basu, Sandip

    2017-01-01

    Three cases of metabolic bone disease in the setting of metastatic neuroendocrine tumor (NET) are illustrated with associated etiopathologies.  One of these cases harbored mixed lesions in the form of vertebral metastasis (biopsy proven) while the other skeletal lesions were caused due to metabolic bone disease related to multiple parathyroid adenomas. While the metastatic lesion was positive on 68Ga-DOTATATE positron emission tomography-computed tomography (PET-CT), the lesions of metabolic bone disease were negative and the 18F-fluoride PET-CT demonstrated the features of metabolic bone scan. Similar picture of metabolic bone disease [18-sodium fluoride (18NaF)/68Ga-DOTATATE mismatch] was documented in the other two patients, while fluorodeoxyglucose (FDG)-PET-CT was variably positive, primarily showing tracer uptake in the metabolic skeletal lesions of the patient with hypersecretion of parathyroid hormone-related protein (PTHrP) by the underlying tumor. Discordance between 18NaF PET-CT and 68Ga-DOTATATE PET-CT serves as a good marker for identification of metabolic bone disease and diagnosing such a clinical entity. In a patient of NET with metabolic bone disease and hypercalcemia, thus, two causes need to be considered: (i) Coexisting parathyroid adenoma in multiple endocrine neoplasia type I (MEN-I) syndrome and (ii) humoral hypercalcemia of malignancy (HHM) related to hypersecretion of PTHrP by the tumor. The correct diagnosis of metabolic bone disease in metastatic NET can alter the management substantially. Interestingly, peptide receptor radionuclide therapy (PRRT) can emerge as a very promising treatment modality in patients of metabolic bone disease caused by HHM in the setting of NET. PMID:28217023

  20. L-lactate transport in Ehrlich ascites-tumour cells.

    PubMed Central

    Spencer, T L; Lehninger, A L

    1976-01-01

    Ehrlich ascites-tumour cells were investigated with regard to their stability to transport L-lactate by measuring either the distribution of [14C]lactate or concomitant H+ ion movements. The movement of lactate was dependent on the pH difference across the cell membrane and was electroneutral, as evidenced by an observed 1:1 antiport for OH- ions or 1:1 symport with H+ ions. 2. Kinetic experiments showed that lactate transport was saturable, with an apparent Km of approx. 4.68 mM and a Vmax. as high as 680 nmol/min per mg of protein at pH 6.2 and 37 degrees C. 3. Lactate transport exhibited a high temperature dependence (activation energy = 139 kJ/mol). 4. Lactate transport was inhibited competitively by (a) a variety of other substituted monocarboxylic acids (e.g. pyruvate, Ki = 6.3 mM), which were themselves transported, (b) the non-transportable analogues alpha-cyano-4-hydroxycinnamate (Ki = 0.5 mM), alpha-cyano-3-hydroxycinnamate (Ki = 2mM) and DL-p-hydroxyphenyl-lactate (Ki = 3.6 mM) and (c) the thiol-group reagent mersalyl (Ki = 125 muM). 5. Transport of simple monocarboxylic acids, including acetate and propionate, was insensitive to these inhibitors; they presumably cross the membrane by means of a different mechanism. 6. Experiments using saturating amounts of mersalyl as an "inhibitor stop" allowed measurements of the initial rates of net influx and of net efflux of [14C]lactate. Influx and efflux of lactate were judged to be symmetrical reactions in that they exhibited similar concentration dependence. 7. It is concluded that lactate transport in Ehrlich ascites-tumour cells is mediated by a carrier capable of transporting a number of other substituted monocarboxylic acids, but not unsubstituted short-chain aliphatic acids. PMID:7237

  1. L-lactate transport in Ehrlich ascites-tumour cells.

    PubMed

    Spencer, T L; Lehninger, A L

    1976-02-15

    Ehrlich ascites-tumour cells were investigated with regard to their stability to transport L-lactate by measuring either the distribution of [14C]lactate or concomitant H+ ion movements. The movement of lactate was dependent on the pH difference across the cell membrane and was electroneutral, as evidenced by an observed 1:1 antiport for OH- ions or 1:1 symport with H+ ions. 2. Kinetic experiments showed that lactate transport was saturable, with an apparent Km of approx. 4.68 mM and a Vmax. as high as 680 nmol/min per mg of protein at pH 6.2 and 37 degrees C. 3. Lactate transport exhibited a high temperature dependence (activation energy = 139 kJ/mol). 4. Lactate transport was inhibited competitively by (a) a variety of other substituted monocarboxylic acids (e.g. pyruvate, Ki = 6.3 mM), which were themselves transported, (b) the non-transportable analogues alpha-cyano-4-hydroxycinnamate (Ki = 0.5 mM), alpha-cyano-3-hydroxycinnamate (Ki = 2mM) and DL-p-hydroxyphenyl-lactate (Ki = 3.6 mM) and (c) the thiol-group reagent mersalyl (Ki = 125 muM). 5. Transport of simple monocarboxylic acids, including acetate and propionate, was insensitive to these inhibitors; they presumably cross the membrane by means of a different mechanism. 6. Experiments using saturating amounts of mersalyl as an "inhibitor stop" allowed measurements of the initial rates of net influx and of net efflux of [14C]lactate. Influx and efflux of lactate were judged to be symmetrical reactions in that they exhibited similar concentration dependence. 7. It is concluded that lactate transport in Ehrlich ascites-tumour cells is mediated by a carrier capable of transporting a number of other substituted monocarboxylic acids, but not unsubstituted short-chain aliphatic acids.

  2. Lactation transcriptomics in the Australian marsupial, Macropus eugenii: transcript sequencing and quantification

    PubMed Central

    Lefèvre, Christophe M; Digby, Matthew R; Whitley, Jane C; Strahm, Yvan; Nicholas, Kevin R

    2007-01-01

    Background Lactation is an important aspect of mammalian biology and, amongst mammals, marsupials show one of the most complex lactation cycles. Marsupials, such as the tammar wallaby (Macropus eugenii) give birth to a relatively immature newborn and progressive changes in milk composition and milk production regulate early stage development of the young. Results In order to investigate gene expression in the marsupial mammary gland during lactation, a comprehensive set of cDNA libraries was derived from lactating tissues throughout the lactation cycle of the tammar wallaby. A total of 14,837 express sequence tags were produced by cDNA sequencing. Sequence analysis and sequence assembly were used to construct a comprehensive catalogue of mammary transcripts. Sequence data from pregnant and early or late lactating specific cDNA libraries and, data from early or late lactation massively parallel sequencing strategies were combined to analyse the variation of milk protein gene expression during the lactation cycle. Conclusion Results show a steady increase in expression of genes coding for secreted protein during the lactation cycle that is associated with high proportion of transcripts coding for milk proteins. In addition, genes involved in immune function, translation and energy or anabolic metabolism are expressed across the lactation cycle. A number of potential new milk proteins or mammary gland remodelling markers, including noncoding RNAs have been identified. PMID:17997866

  3. Lactate in bitches with pyometra.

    PubMed

    Volpato, R; Rodello, L; Abibe, R B; Lopes, M D

    2012-12-01

    Lactate is a compound produced by the anaerobic metabolism of glucose, and hyperlactataemia occurs when the rate of production of lactate exceeds the rate of elimination. This occurs in situations of hypoxia and tissue hypoperfusion. Lactate has been considered a useful prognostic indicator in critically ill patients. Pyometra is a disease of adult female dogs characterized by inflammation of the uterus with an accumulation of exudate, which occurs during the luteal phase. It is one of the most common diseases that occur in the genital tract of female dogs. A total of 31 dogs were diagnosed with pyometra. The diagnosis was confirmed at ultrasonography. Of the 31 dogs, 25 females had open cervix pyometra and six had closed cervix pyometra. Plasma lactate concentrations were determined by an enzymatic colorimetric method. The average concentration (±SD) of plasma lactate in all 31 bitches with pyometra was 3.55 ± 0.46 mm. Healthy dogs had plasma lactate concentrations between 0.3 and 2.5 mm (mean ± SD). Concentrations ranged from 0.8 to 2.9 mm when plasma lactate was measured with a portable device and 0.4-2.6 mm with the blood gas analyser. Even though plasma lactate values vary between several studies and equipment used to measure concentrations, our results for dogs with pyometra are higher indicating hyperlactataemia (Thorneloe et al. , Can Vet J 48, 283-288). Plasma lactate in dogs with closed cervix pyometra was mean ± SD and in dogs with open cervix pyometra, it was mean ± SD. The plasma lactate concentration in dogs with pyometra was higher than in healthy bitches, and there was no influence of patency of the cervix on the concentration of plasma lactate concentrations. Plasma lactate concentrations were similar for animals with open and closed pyometra (3.54 ± 0.52 to 3.64 ± 1.03 mm).

  4. Expression of lactate dehydrogenase C correlates with poor prognosis in renal cell carcinoma.

    PubMed

    Hua, Yibo; Liang, Chao; Zhu, Jundong; Miao, Chenkui; Yu, Yajie; Xu, Aimin; Zhang, Jianzhong; Li, Pu; Li, Shuang; Bao, Meiling; Yang, Jie; Qin, Chao; Wang, Zengjun

    2017-03-01

    Lactate dehydrogenase C is an isoenzyme of lactate dehydrogenase and a member of the cancer-testis antigens family. In this study, we aimed to investigate the expression and functional role of lactate dehydrogenase C and its basic mechanisms in renal cell carcinoma. First, a total of 133 cases of renal cell carcinoma samples were analysed in a tissue microarray, and Kaplan-Meier survival curve analyses were performed to investigate the correlation between lactate dehydrogenase C expression and renal cell carcinoma progression. Lactate dehydrogenase C protein levels and messenger RNA levels were significantly upregulated in renal cell carcinoma tissues, and the patients with positive lactate dehydrogenase C expression had a shorter progression-free survival, indicating the oncogenic role of lactate dehydrogenase C in renal cell carcinoma. In addition, further cytological experiments demonstrated that lactate dehydrogenase C could prompt renal cell carcinoma cells to produce lactate, and increase metastatic and invasive potential of renal cell carcinoma cells. Furthermore, lactate dehydrogenase C could induce the epithelial-mesenchymal transition process and matrix metalloproteinase-9 expression. In summary, these findings showed lactate dehydrogenase C was associated with poor prognosis in renal cell carcinoma and played a pivotal role in the migration and invasion of renal cell carcinoma cells. Lactate dehydrogenase C may act as a novel biomarker for renal cell carcinoma progression and a potential therapeutic target for the treatment of renal cell carcinoma.

  5. Parathyroid hormone-related protein induces hypertrophy in podocytes via TGF-beta(1) and p27(Kip1): implications for diabetic nephropathy.

    PubMed

    Romero, Montserrat; Ortega, Arantxa; Izquierdo, Adriana; López-Luna, Pilar; Bosch, Ricardo J

    2010-08-01

    Hypertrophy of podocytes is characteristic in diabetic nephropathy (DN). Previously, we observed the upregulation of parathyroid hormone-related protein (PTHrP) and its receptor PTH1R, in experimental DN, associated with renal hypertrophy. Herein, we test the hypothesis that PTHrP participates in the mechanism of high glucose (HG)-induced podocyte hypertrophy. On mouse podocytes, hypertrophy was assessed by protein content/cell and [H(3)]leucine incorporation. Podocytes were stimulated with HG (25 mM), PTHrP(1-36) (100 nM), angiotensin II (AngII) (100 nM) or TGF-beta(1) (5 ng/mL) in the presence or absence of PTHrP-neutralizing antibodies (alpha-PTHrP), the PTH1R antagonist JB4250 (10 microM), PTHrP silencer RNA (siRNA) or TGF-beta(1) siRNA. Protein expression was analysed by western blot and immunohistochemistry. HG-induced hypertrophy was abolished in the presence of either alpha-PTHrP or PTHrP siRNA. This effect was associated with an inhibition of the upregulation of TGF-beta(1) and p27(Kip1). JB4250 also inhibited HG-induced p27(Kip1) upregulation. Interestingly, whilst HG and AngII were unable to stimulate the expression of p27(Kip1) on PTHrP siRNA-transfected podocytes, TGF-beta(1) was still able to upregulate p27(Kip1) in these cells. Moreover, HG and PTHrP-induced hypertrophy as well as p27(Kip1) upregulation were abolished on TGF-beta(1) siRNA-transfected podocytes. Furthermore, the glomeruli of transgenic PTHrP-overexpressing mice showed a constitutive overexpression of TGF-beta(1) and p27(Kip1) to a degree similar to that of diabetic animals. PTHrP seems to participate in the hypertrophic signalling triggered by HG. In this condition, AngII induces the upregulation of PTHrP, which might induce the expression of TGF-beta(1) and p27(Kip1). These findings provide new insights into the protective effects of AngII antagonists in DN, opening new paths for intervention.

  6. Lactate modulates the activity of primary cortical neurons through a receptor-mediated pathway.

    PubMed

    Bozzo, Luigi; Puyal, Julien; Chatton, Jean-Yves

    2013-01-01

    Lactate is increasingly described as an energy substrate of the brain. Beside this still debated metabolic role, lactate may have other effects on brain cells. Here, we describe lactate as a neuromodulator, able to influence the activity of cortical neurons. Neuronal excitability of mouse primary neurons was monitored by calcium imaging. When applied in conjunction with glucose, lactate induced a decrease in the spontaneous calcium spiking frequency of neurons. The effect was reversible and concentration dependent (IC50 ∼4.2 mM). To test whether lactate effects are dependent on energy metabolism, we applied the closely related substrate pyruvate (5 mM) or switched to different glucose concentrations (0.5 or 10 mM). None of these conditions reproduced the effect of lactate. Recently, a Gi protein-coupled receptor for lactate called HCA1 has been introduced. To test if this receptor is implicated in the observed lactate sensitivity, we incubated cells with pertussis toxin (PTX) an inhibitor of Gi-protein. PTX prevented the decrease of neuronal activity by L-lactate. Moreover 3,5-dyhydroxybenzoic acid, a specific agonist of the HCA1 receptor, mimicked the action of lactate. This study indicates that lactate operates a negative feedback on neuronal activity by a receptor-mediated mechanism, independent from its intracellular metabolism.

  7. Lactate Modulates the Activity of Primary Cortical Neurons through a Receptor-Mediated Pathway

    PubMed Central

    Bozzo, Luigi; Puyal, Julien; Chatton, Jean-Yves

    2013-01-01

    Lactate is increasingly described as an energy substrate of the brain. Beside this still debated metabolic role, lactate may have other effects on brain cells. Here, we describe lactate as a neuromodulator, able to influence the activity of cortical neurons. Neuronal excitability of mouse primary neurons was monitored by calcium imaging. When applied in conjunction with glucose, lactate induced a decrease in the spontaneous calcium spiking frequency of neurons. The effect was reversible and concentration dependent (IC50 ∼4.2 mM). To test whether lactate effects are dependent on energy metabolism, we applied the closely related substrate pyruvate (5 mM) or switched to different glucose concentrations (0.5 or 10 mM). None of these conditions reproduced the effect of lactate. Recently, a Gi protein-coupled receptor for lactate called HCA1 has been introduced. To test if this receptor is implicated in the observed lactate sensitivity, we incubated cells with pertussis toxin (PTX) an inhibitor of Gi-protein. PTX prevented the decrease of neuronal activity by L-lactate. Moreover 3,5-dyhydroxybenzoic acid, a specific agonist of the HCA1 receptor, mimicked the action of lactate. This study indicates that lactate operates a negative feedback on neuronal activity by a receptor-mediated mechanism, independent from its intracellular metabolism. PMID:23951229

  8. False-positive postmortem EMIT drugs-of-abuse assay due to lactate dehydrogenase and lactate in urine.

    PubMed

    Sloop, G; Hall, M; Simmons, G T; Robinson, C A

    1995-01-01

    Three cases of multiple false-positive drug tests are described. Postmortem urine specimens were screened using the enzyme-multiplied immunoassay technique. All patients had proteinuria and lactic aciduria. These false-positive reactions were due to the presence of lactate dehydrogenase (LDH), lactic acid, and protein. This finding was confirmed by creating a multiple false-positive sample with a solution of LDH and lactate in 5% bovine serum albumin at pH 6.

  9. Lipid-Based Nutrient Supplements During Pregnancy and Lactation Did Not Affect Human Milk Oligosaccharides and Bioactive Proteins in a Randomized Trial.

    PubMed

    Jorgensen, Josh M; Arnold, Charles; Ashorn, Per; Ashorn, Ulla; Chaima, David; Cheung, Yin Bun; Davis, Jasmine Cc; Fan, Yue-Mei; Goonatilleke, Elisha; Kortekangas, Emma; Kumwenda, Chiza; Lebrilla, Carlito B; Maleta, Kenneth; Totten, Sarah M; Wu, Lauren D; Dewey, Kathryn G

    2017-08-09

    Background: Human milk oligosaccharides (HMOs) and bioactive proteins are beneficial to infant health. Recent evidence suggests that maternal nutrition may affect the amount of HMOs and proteins in breast milk; however, the effect of nutrient supplementation on HMOs and bioactive proteins has not yet been well studied.Objective: We aimed to determine whether lipid-based nutrient supplements (LNSs) affect milk bioactive protein and HMO concentrations at 6 mo postpartum in women in rural Malawi. These are secondary outcomes of a previously published randomized controlled trial.Methods: Women were randomly assigned to consume either an iron and folic acid capsule (IFA) daily from ≤20 wk gestation until delivery, followed by placebo daily from delivery to 6 mo postpartum, or a multiple micronutrient (MMN) capsule or LNS daily from ≤20 wk gestation to 6 mo postpartum. Breast milk concentrations of total HMOs, sialylated HMOs, fucosylated HMOs, lactoferrin, lactalbumin, lysozymes, antitrypsin, immunoglobulin A, and osteopontin were analyzed at 6 mo postpartum (n = 647). Between-group differences in concentrations and in proportions of women classified as having low concentrations were tested.Results: HMO and bioactive protein concentrations did not differ between groups (P > 0.10 for all comparisons). At 6 mo postpartum, the proportions of women with low HMOs or bioactive proteins were not different between groups except for osteopontin. A lower proportion of women in the IFA group had low osteopontin compared with the LNS group after adjusting for covariates (OR: 0.5; 95% CI: 0.3, 0.9; P = 0.016).Conclusion: The study findings do not support the hypothesis that supplementation with an LNS or MMN capsule during pregnancy and postpartum would increase HMO or bioactive milk proteins at 6 mo postpartum among Malawian women. This trial was registered at clinicaltrials.gov as NCT01239693. © 2017 American Society for Nutrition.

  10. Contraceptive implants and lactation.

    PubMed

    Díaz, Soledad

    2002-01-01

    The safety and efficacy of four contraceptive implants, plant, Implanon, Nestorone and Elcometrine, have been evaluated during use in the postpartum period by lactating women. These implants provide highly effective contraceptive protection with no negative effect on breastfeeding or infant growth and development. Breastfeeding women initiating Norplant use in the second postpartum month experience significantly longer periods of amenorrhea than do untreated women or intrauterine device users. After weaning, the bleeding pattern is similar to that observed in non-nursing women. Norplant use does not affect bone turnover and density during lactation. Norplant and Implanon release orally active progestins while Nestorone and Elcometrine implants release an orally inactive progestin, which represents an advantage since the infant should be free of steroidal effects. The infant's daily intake of steroids (estimated from concentrations in maternal milk during the first month of use) range from 90 to 100 ng of levonorgestrel (Norplant), 75-120 ng of etonogestrel (Implanon), and 50 ng and 110 ng of Nestorone (Nestorone and Elcometrine implants, respectively). Nursing women needing contraception may use progestin-only implants when nonhormonal methods are not available or acceptable. Implants that deliver orally active steroids should only be used after 6 weeks postpartum to avoid transferring of steroids to the newborn.

  11. Partial reconstruction of in vitro gluconeogenesis arising from mitochondrial l-lactate uptake/metabolism and oxaloacetate export via novel L-lactate translocators.

    PubMed Central

    De Bari, Lidia; Atlante, Anna; Valenti, Daniela; Passarella, Salvatore

    2004-01-01

    In the light of the occurrence of L-lactate dehydrogenase inside the mitochondrial matrix, we looked at whether isolated rat liver mitochondria can take up and metabolize L-lactate, and provide oxaloacetate outside mitochondria, thus contributing to a partial reconstruction of gluconeogenesis in vitro. We found that: (1) L-lactate (10 mM), added to mitochondria in the presence of a cocktail of glycolysis/gluconeogenesis enzymes and cofactors, can lead to synthesis of glyceraldehyde-3-phosphate at a rate of about 7 nmol/min per mg mitochondrial protein. (2) Three novel translocators exist to mediate L-lactate traffic across the inner mitochondrial membrane. An L-lactate/H+ symporter was identified by measuring fluorimetrically the rate of endogenous pyridine nucleotide reduction. Consistently, L-lactate oxidation was found to occur with P/O ratio=3 (where P/O ratio is the ratio of mol of ATP synthesized to mol of oxygen atoms reduced to water during oxidative phosphorylation) and with generation of membrane potential. Proton uptake, which occurred as a result of addition of L-lactate to RLM together with electron flow inhibitors, and mitochondrial swelling in ammonium L-lactate solutions were also monitored. L-Lactate/oxaloacetate and L-lactate/pyruvate anti-porters were identified by monitoring photometrically the appearance of L-lactate counter-anions outside mitochondria. These L-lactate translocators, which are distinct from the monocarboxylate carrier, were found to differ from each other in V(max) values and in inhibition and pH profiles, and proved to regulate mitochondrial L-lactate metabolism in vitro. The role of lactate/mitochondria interactions in gluconeogenesis is discussed. PMID:14960150

  12. Diet-induced obesity accelerates blood lactate accumulation of rats in response to incremental exercise to maximum.

    PubMed

    Chen, Chiao-Nan Joyce; Liao, Yi-Hung; Lin, Shang-Ying; Yu, Jun-Xian; Li, Zhen-Jie; Lin, Yu-Chieh; Chang, Gwo-Jyh; Lin, Chung-Hao; Wong, Alice May-Kuen

    2017-08-30

    Blood lactate increases during incremental exercise at high-intensity workloads and limited exercise capacity is a characteristic of obese animals. This study examined whether blood lactate changes in response to incremental exercise is disrupted in obese animals. Muscular and hepatic proteins that are critical in lactate metabolism were also investigated. Rats were randomized to either standard chow (control) or high fat diet (HFD) groups. All animals underwent an incremental treadmill test after 14 weeks of diet intervention. Blood lactate levels were measured before and after the treadmill test. Activities of mitochondrial oxidative phosphorylation and glycolysis were examined in muscle tissues. Proteins in the liver and skeletal muscles that participate in the turnover of blood lactate were determined by Western blot. Running time in the incremental treadmill test decreased in the HFD group, and blood lactate accumulated faster in these animals than in the control group. Animals with HFD had a decreased level of hepatic monocarboxylate transporter 2, the protein responsible for blood lactate uptake in the liver. Skeletal muscles of animals with HFD showed greater glycolytic activity and decreased content of lactate dehydrogenase B, which converts lactate to pyruvate. Blood lactate accumulated faster during incremental exercise in obese animals and associated with their decreased exercise performance. Changes in the metabolic pattern of muscles and changes of liver and muscle proteins associated with lactate utilization likely contribute to the abnormal response of blood lactate to incremental exercise in obese animals. Copyright © 2017, American Journal of Physiology-Regulatory, Integrative and Comparative Physiology.

  13. Interactive Effect of Corticosterone and Lactate on Regulation of Testosterone Production in Rat Leydig Cells.

    PubMed

    Chen, Chih-Chieh; Chen, Chien-Wei; Lin, Po-Han; Chou, Jou-Chun; Weng, Ting-Chun; Jian, Cai-Yun; Hu, Sindy; Lai, Wei-Ho; Lieu, Fu-Kong; Wang, Shyi-Wu; Wang, Paulus S

    2017-08-01

    The increasing intensity of exercise enhanced corticosterone and lactate production in both humans and rodents. Our previous studies also demonstrated that lactate could stimulate testosterone production in vivo and in vitro. However, the production of testosterone in response to combined corticosterone and lactate on Leydig cells, and underlying molecular mechanisms are remained unclear. This study investigated the changes in testosterone levels of Leydig cells upon exposure to lactate, corticosterone or combination of both, and revealed the detailed mechanisms. Leydig cells were isolated from rat testes, and treated with different concentrations of lactate (2.5-20 mM), cortiosterone (10(-9) -10(-4)  M) and lactate plus corticosterone. The production of testosterone were assayed by radioimmunoassay, and the key molecular proteins, including luteinizing hormone receptor (LHR), protein kinase A (PKA), steroidogenic acute regulatory protein (StAR), and cholesterol P450 side-chain cleavage enzyme (P450scc) involved in testosterone production were performed by Western blot. Results showed that testosterone levels were significantly increased with lactate, while decresed with corticosterone and lactate plus corticosterone treatment. Protein expressions of LHR and P450scc were upregulated with lactate treatment. However, PKA and P450scc were downregulated by lactate plus corticosterone treatment. This downregulation was followed by decreased testoterone levels in Leydig cells. Furthermore, acetylated cAMP, which activates testosterone production was increased with lactate, but not altered by conrtiosterone. Our findings conclude that corticosterone may interfere with lactate, and restrict lactate-stimulated testosterone production in Leydig cells. J. Cell. Physiol. 232: 2135-2144, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  14. Temporarily decreasing progesterone after timed artificial insemination decreased expression of interferon-tau stimulated gene 15 (ISG15) in blood leukocytes, serum pregnancy-specific protein B concentrations, and embryo size in lactating Holstein cows.

    PubMed

    Carvalho, P D; Consentini, C C; Weaver, S R; Barleta, R V; Hernandez, L L; Fricke, P M

    2017-04-01

    Our objective was to evaluate the effects of temporarily decreasing progesterone (P4) after timed artificial insemination (TAI) on embryonic growth in dairy cows. Lactating Holstein cows (n = 80) were submitted to a Double-Ovsynch protocol for first TAI and were assigned randomly to receive 12.5 mg of PGF2α 5 d after the last GnRH treatment (LowP4) or remain untreated (control). Blood samples were collected thrice weekly from 5 to 29 d after TAI for all cows and from 32 to 67 d for pregnant cows, and were analyzed for P4 and pregnancy-specific protein B concentrations. Expression of interferon-tau stimulated gene 15 (ISG15) was assessed in blood leukocyte mRNA 18 and 20 d after TAI. Pregnancy diagnosis was performed weekly using ultrasound from 32 to 67 d after TAI, and embryonic crown-rump length was measured 32, 39, and 46 d after TAI. Data were analyzed by ANOVA and logistic regression using the MIXED and GLIMMIX procedures of SAS. The LowP4 cows had less P4 than control cows from 6 to 11 d after TAI; however, pregnancy outcomes 32 d after TAI and pregnancy loss from 32 to 67 d after TAI did not differ between treatments. Control cows diagnosed pregnant 32 d after TAI had greater expression of ISG15 20 d after TAI than LowP4 cows diagnosed pregnant 32 d after TAI, and pregnant control cows had greater pregnancy-specific protein B concentrations from 25 to 67 d after TAI than pregnant LowP4 cows. Embryo size did not differ between treatments 32 and 39 d after TAI, but control cows had larger embryos 46 d after TAI. In conclusion, temporarily decreasing P4 after TAI decreased embryonic growth during early pregnancy in lactating Holstein cows but did not affect pregnancies per artificial insemination or pregnancy loss. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  15. Short-term administration of rhGH increases markers of cellular proliferation, but not milk protein gene expression in normal lactating women.

    USDA-ARS?s Scientific Manuscript database

    Growth hormone is one of few pharmacologic agents known to augment milk production in humans. We hypothesized that recombinant human GH (rhGH) increases the expression of cell proliferation and milk protein synthesis genes. Sequential milk and blood samples collected over four days were obtained fro...

  16. Supplementing rumen-protected methionine and lysine in low-protein diets based on corn distillers grains fed to lactating dairy cows

    USDA-ARS?s Scientific Manuscript database

    Feeding rumen-protected methionine (RPM) and lysine (RPL) may allow feeding lower crude protein (CP) diets to dairy cows, thereby increasing nitrogen efficiency and reducing environmental impact. Moreover, RPL supplementation may improve the value of corn distillers dried grains plus solubles (DDGS)...

  17. Local delivery of parathyroid hormone-related protein-derived peptides coated onto a hydroxyapatite-based implant enhances bone regeneration in old and diabetic rats.

    PubMed

    Ardura, Juan A; Portal-Núñez, Sergio; Lozano, Daniel; Gutiérrez-Rojas, Irene; Sánchez-Salcedo, Sandra; López-Herradón, Ana; Mulero, Francisca; Villanueva-Peñacarrillo, María L; Vallet-Regí, María; Esbrit, Pedro

    2016-08-01

    Diabetes mellitus (DM) and aging are associated with bone fragility and increased fracture risk. Both (1-37) N- and (107-111) C-terminal parathyroid hormone-related protein (PTHrP) exhibit osteogenic properties. We here aimed to evaluate and compare the efficacy of either PTHrP (1-37) or PTHrP (107-111) loaded into gelatin-glutaraldehyde-coated hydroxyapatite (HA-Gel) foams to improve bone repair of a transcortical tibial defect in aging rats with or without DM, induced by streptozotocin injection at birth. Diabetic old rats showed bone structural deterioration compared to their age-matched controls. Histological and μ-computerized tomography studies showed incomplete bone repair at 4 weeks after implantation of unloaded Ha-Gel foams in the transcortical tibial defects, mainly in old rats with DM. However, enhanced defect healing, as shown by an increase of bone volume/tissue volume and trabecular and cortical thickness and decreased trabecular separation, occurred in the presence of either PTHrP peptide in the implants in old rats with or without DM. This was accompanied by newly formed bone tissue around the osteointegrated HA-Gel implant and increased gene expression of osteocalcin and vascular endothelial growth factor (bone formation and angiogenic markers, respectively), and decreased expression of Sost gene, a negative regulator of bone formation, in the healing bone area. Our findings suggest that local delivery of PTHrP (1-37) or PTHrP (107-111) from a degradable implant is an attractive strategy to improve bone regeneration in aged and diabetic subjects. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2060-2070, 2016.

  18. Inhibition of terminal chondrocyte differentiation by bone morphogenetic protein 7 (OP-1) in vitro depends on the periarticular region but is independent of parathyroid hormone-related peptide.

    PubMed

    Haaijman, A; Karperien, M; Lanske, B; Hendriks, J; Löwik, C W; Bronckers, A L; Burger, E H

    1999-10-01

    Bone morphogenetic protein-7, or BMP-7 (OP-1), is highly expressed in the perichondrium of embryonic long bones and is thought to play a role in endochondral ossification. Previously we have shown that BMP-7 inhibits terminal chondrocyte differentiation; that is, chondrocyte hypertrophy and mineralization in cultured explants of embryonic mouse metatarsals. However, the mechanism of this inhibition and the target cells of BMP-7 are still unknown. In this study we show that BMP-7 inhibits terminal chondrocyte differentiation indirectly, via an interaction with the periarticular region of the explants. This region also expresses parathyroid hormone-related peptide (PTHrP). PTHrP regulates terminal chondrocyte differentiation by inhibiting hypertrophic differentiation of prehypertrophic chondrocytes. The differentiating center in turn regulates PTHrP expression via a feedback loop involving Indian hedgehog (Ihh), which is expressed in the prehypertrophic chondrocytes. Ihh is thought to act on perichondrial cells, which in turn start to express an as yet unknown mediator that stimulates PTHrP expression in the periarticular region. It has been suggested that this factor belongs to the BMP-family. We investigated whether the inhibition of terminal chondrocyte differentiation by BMP-7 was due to upregulation of the PTHrP-Ihh feedback loop and whether BMP-7 was the unknown factor in the loop. Here we show that exogenous BMP-7 did not upregulate the mRNA expression of PTHrP, Ihh, or the PTH/PTHrP receptor in cultured wild-type embryonic metatarsals. Furthermore, BMP-7 could still inhibit terminal chondrocyte differentiation in the metatarsals of PTHrP-deficient (PTHrP-/-) mouse embryos. These data indicate that the BMP-7-mediated inhibition of terminal chondrocyte differentiation in vitro is independent of the PTHrP-Ihh feedback loop. We concluded that BMP-7 modulates terminal chondrocyte differentiation and cartilage mineralization of fetal bone explants in vitro via as

  19. Tumour-derived PTH-related protein triggers adipose tissue browning and cancer cachexia.

    PubMed

    Kir, Serkan; White, James P; Kleiner, Sandra; Kazak, Lawrence; Cohen, Paul; Baracos, Vickie E; Spiegelman, Bruce M

    2014-09-04

    Cachexia is a wasting disorder of adipose and skeletal muscle tissues that leads to profound weight loss and frailty. About half of all cancer patients suffer from cachexia, which impairs quality of life, limits cancer therapy and decreases survival. One key characteristic of cachexia is higher resting energy expenditure levels than in healthy individuals, which has been linked to greater thermogenesis by brown fat. How tumours induce brown fat activity is unknown. Here, using a Lewis lung carcinoma model of cancer cachexia, we show that tumour-derived parathyroid-hormone-related protein (PTHrP) has an important role in wasting, through driving the expression of genes involved in thermogenesis in adipose tissues. Neutralization of PTHrP in tumour-bearing mice blocked adipose tissue browning and the loss of muscle mass and strength. Our results demonstrate that PTHrP mediates energy wasting in fat tissues and contributes to the broader aspects of cancer cachexia. Thus, neutralization of PTHrP might hold promise for ameliorating cancer cachexia and improving patient survival.

  20. Plasma D-lactate levels in diagnosis of appendicitis.

    PubMed

    Cağlayan, Fatma; Cakmak, Murat; Cağlayan, Osman; Cavuşoglu, Turgut

    2003-01-01

    We investigated the possible use of D-lactate as a predictor in the diagnosis of appendicitis. C-reactive protein level (CRP) and leukocyte counts were also evaluated. Venous blood D-lactate, CRP, and leukocyte counts were measured preoperatively in 53 patients undergoing surgery for appendicitis, as well as in 20 healthy subjects. Levels of all three parameters in the surgical patients were significantly higher than in the control group (p <.05). Previous studies have shown that venous D-lactate is more specific to the intestine than CPR or leukocyte count. Based on our data, venous D-lactate, which had the lowest false-negative rate among these laboratory parameters, may be a useful diagnostic marker for appendicitis. None of these parameters were helpful in identifying the type of the appendicitis.

  1. Involvement of parathyroid hormone-related protein in vascular calcification of chronic haemodialysis patients.

    PubMed

    Liu, Fang; Fu, Ping; Fan, Wenxing; Gou, Rong; Huang, Youqun; Qiu, Hongyu; Zhong, Hui; Huang, Songmin

    2012-08-01

    To investigate the role of parathyroid hormone-related protein (PTHrP) in vascular calcification of patients with chronic hemodialysis. The inferior epigastric arteries were obtained from 23 patients on chronic haemodialysis and 16 patients with renal carcinoma as control. Haematoxylin-eosin staining, elastic fibre staining, Alizarin Red calcium staining and immunohistochemical staining of PTHrP, bone morphogenetic protein-2 (BMP-2), Cbfa1/Runx2 were performed. Real-time polymerase chain reaction (PCR) was used to examine mRNA expressions of PTHrP, BMP-2 and Cbfa1/Runx2. Western blot and real-time PCR were used to detect the effects of PTHrP-siRNA and rh-PTHrP-1-34 on the expressions of PTHrP, BMP-2 and Cbfa1/Runx2 in human aortic smooth muscle cells (HASMC). Alkaline phosphatase (ALP) activities and intracellular calcium content in HASMCs were assessed after treatment with 10 mmol/L β-glycerol phosphoric acid for 48 h. Vascular calcification was confirmed in 78.2% of patients on chronic haemodialysis, and the expressions of PTHrP, BMP-2 and Cbfa1 in the arteries were significantly upregulated. PTHrP-siRNA could downregulate the expression of PTHrP by 60%, BMP-2 by 25% and Cbfa1 by 25% at 24 h (P < 0.05). Exogenous rh-PTHrP-1-34 could upregulate the expressions of BMP-2 and Cbfa1 by 1.37-fold and 1.46-fold, respectively, at 24 h in a time-independent manner (P < 0.05), which were attenuated by PTHrP-siRNA. Moreover, it could promote intracellular calcium deposition and increase ALP activities, which were partially blocked by PTHrP-siRNA (P < 0.05). Vascular calcification was common in patients with chronic haemodialysis, to which PTHrP might contribute by activating BMP-2/ Cbfa1 signalling pathway. © 2012 The Authors. Nephrology © 2012 Asian Pacific Society of Nephrology.

  2. Jussara (Euterpe edulis Mart.) Supplementation during Pregnancy and Lactation Modulates the Gene and Protein Expression of Inflammation Biomarkers Induced by trans-Fatty Acids in the Colon of Offspring

    PubMed Central

    Almeida Morais, Carina; Oyama, Lila Missae; de Oliveira, Juliana Lopez; Carvalho Garcia, Márcia; de Rosso, Veridiana Vera; Sousa Mendes Amigo, Laís; do Nascimento, Claudia Maria Oller; Pisani, Luciana Pellegrini

    2014-01-01

    Maternal intake of trans-fatty acids (TFAs) in the perinatal period triggers a proinflammatory state in offspring. Anthocyanins contained in fruit are promising modulators of inflammation. This study investigated the effect of Jussara supplementation in the maternal diet on the proinflammatory state of the colon in offspring exposed to perinatal TFAs. On the first day of pregnancy rats were divided into four groups: control diet (C), control diet with 0.5% Jussara supplementation (CJ), diet enriched with hydrogenated vegetable fat, rich in TFAs (T), or T diet supplemented with 0.5% Jussara (TJ) during pregnancy and lactation. We showed that Jussara supplementation in maternal diet (CJ and TJ groups) reduced carcass lipid/protein ratios, serum lipids, glucose, IL-6, TNF-α, gene expression of IL-6R, TNF-αR (P < 0.05), TLR-4 (P < 0.01), and increase Lactobacillus spp. (P < 0.05) in the colon of offspring compared to the T group. The IL-10 (P = 0.035) and IL-10/TNF-α ratio (P < 0.01) was higher in the CJ group than in the T group. The 0.5% Jussara supplementation reverses the adverse effects of perinatal TFAs, improving lipid profiles, glucose levels, body composition, and gut microbiota and reducing low-grade inflammation in the colon of 21-day-old offspring, and could contribute to reducing chronic disease development. PMID:25276060

  3. Maternal protein restriction during gestation and lactation in the rat results in increased brain levels of kynurenine and kynurenic acid in their adult offspring.

    PubMed

    Honório de Melo Martimiano, Paula; de Sa Braga Oliveira, André; Ferchaud-Roucher, Véronique; Croyal, Mikaël; Aguesse, Audrey; Grit, Isabelle; Ouguerram, Khadija; Lopes de Souza, Sandra; Kaeffer, Bertrand; Bolaños-Jiménez, Francisco

    2017-01-01

    Early malnutrition is a risk factor for depression and schizophrenia. Since the offspring of malnourished dams exhibit increased brain levels of serotonin (5-HT), a tryptophan-derived neurotransmitter involved in the pathophysiology of these mental disorders, it is believed that the deleterious effects of early malnutrition on brain function are due in large part to altered serotoninergic neurotransmission resulting from impaired tryptophan (Trp) metabolism. However, tryptophan is also metabolized through the kynurenine (KYN) pathway yielding several neuroactive compounds including kynurenic (KA), quinolinic (QA) and xanthurenic (XA) acids. Nevertheless, the impact of perinatal malnutrition on brain kynurenine pathway metabolism has not been examined to date. Here, we used ultra-performance liquid chromatography-tandem mass spectrometry for the simultaneous quantification of tryptophan and a set of seven compounds spanning its metabolism through the serotonin and kynurenine pathways, in the brain of embryos and adult offspring of rat dams fed a protein-restricted (PR) diet. Protein-restricted embryos showed reduced brain levels of Trp, serotonin and KA, but not of KYN, XA, or QA. In contrast, PR adult rats exhibited enhanced levels of Trp in the brainstem and cortex along with increased concentrations of 5-HT, kynurenine and XA. The levels of XA and KA were also increased in the hippocampus of adult PR rats. These results show that early protein deficiency induces selective and long-lasting changes in brain kynurenine metabolism. Given the regulatory role of KYN pathway metabolites on brain development and function, these changes might contribute to the risk of developing psychiatric disorders induced by early malnutrition. © 2016 International Society for Neurochemistry.

  4. Effect of chronic thyroxine treatment on IGF-I, IGF-II and IGF-binding protein expression in mammary gland and liver during pregnancy and early lactation in rats.

    PubMed

    Rosato, Roberto; Lindenbergh-Kortleve, Dicky; Neck, Johan van; Drop, Stenvert; Jahn, Graciela

    2002-05-01

    Hyperthyroidism in rats produces organ hypertrophy and increases in circulating IGF-I and IGF-binding protein (IGFBP)-3. Chronic treatment with thyroxine (T(4)) during pregnancy advances parturition, blocks lactation and changes several hormone receptors in mammary gland and liver. Since IGFs are implicated in mammary and liver growth and in differentiation, we studied the effects of hyperthyroidism, induced by daily injections of T(4) (0.25 mg/kg). Using quantitative RT-PCR and in situ hybridization, the gene expression of IGF-I, IGF-II and the IGFBPs was determined in mammary gland and liver of rats at estrus and days 7, 14 and 21 of pregnancy (G7, G14, G21), day 1 postpartum (L1) and 3 days after removing the litter (L4). Circulating levels of IGF-I, tri-iodothyronine (T(3)), PRL and GH were measured. T(4) treatment (HT) increased circulating T(3) save on G21, did not change serum IGF-I, increased PRL on G21 and decreased GH on L1. PRL decreased on L1 because of the absence of lactation. Hepatic IGF-I mRNA was low during pregnancy and increased on L4. HT advanced this increase to L1. In controls, liver IGFBP-3 mRNA levels decreased from G14 to G21, whereas IGFBP-4 showed an inverse pattern. HT lowered IGFBP-3 mRNA and increased IGFBP-4. Increases in mammary concentrations of IGF-I, IGFBP-3 and IGFBP-4 mRNAs were seen on G21. HT delayed these peaks to L1. Mammary IGF-II and IGFBP-2 mRNA levels were high on G7 and G14, and fell afterwards, with HT having no effects. IGFBP-5 mRNA decreased during pregnancy and increased on L1. HT increased IGFBP-5 levels in early pregnancy and on L1. IGF-I mRNA localized to connective and epithelial mammary tissue, while IGFBP-2 and IGFBP-5 mRNA was only in epithelial cells. These results imply a role for IGF-I, IGFBP-3 and IGFBP-4 in terminal mammary development, while IGF-II and IGFBP-2 may be implicated in early growth. IGFBP-5 has been implicated in mammary apoptosis, and the HT-induced increase may play a role in the premature

  5. Utility of inline milk fat and protein ratio to diagnose subclinical ketosis and to assign propylene glycol treatment in lactating dairy cows

    PubMed Central

    Jenkins, Nicholas T.; Peña, Gustavo; Risco, Carlos; Barbosa, Carolina C.; Vieira-Neto, Achilles; Galvão, Klibs N.

    2015-01-01

    The objective was to identify a fat-to-protein ratio (FPR) cut-off to diagnose subclinical ketosis (SCK) and to evaluate the effect of propylene glycol (PPG) treatment of cows with high FPR. The optimized cut-off was > 1.42; sensitivity (Se) = 92%; specificity (Sp) = 65%. A cut-off > 1.5 was selected for the PPG trial for balanced Se-Sp. Fat-to-protein ratio cut-offs > 1.25, 1.35, 1.50, 1.60, and 1.70 resulted in Se-Sp of 100% to 49%, 96% to 59%, 75% to 78%, 33% to 90%, and 8% to 96%, respectively. The proportions of cows with FPR > 1.25, 1.35, 1.42, 1.50, 1.60, and 1.70 were 60%, 50%, 44%, 30%, 14%, and 6%, respectively. Incidences of clinical ketosis and milk yield were similar between cows that received 400 mL of PPG (n = 34) and control cows (n = 38). Prevalence of SCK at enrollment was 29.2%; therefore, FPR > 1.5 is not indicated for treatment. Lower cut-offs should be used for screening. PMID:26246632

  6. Utility of inline milk fat and protein ratio to diagnose subclinical ketosis and to assign propylene glycol treatment in lactating dairy cows.

    PubMed

    Jenkins, Nicholas T; Peña, Gustavo; Risco, Carlos; Barbosa, Carolina C; Vieira-Neto, Achilles; Galvão, Klibs N

    2015-08-01

    The objective was to identify a fat-to-protein ratio (FPR) cut-off to diagnose subclinical ketosis (SCK) and to evaluate the effect of propylene glycol (PPG) treatment of cows with high FPR. The optimized cut-off was > 1.42; sensitivity (Se) = 92%; specificity (Sp) = 65%. A cut-off > 1.5 was selected for the PPG trial for balanced Se-Sp. Fat-to-protein ratio cut-offs > 1.25, 1.35, 1.50, 1.60, and 1.70 resulted in Se-Sp of 100% to 49%, 96% to 59%, 75% to 78%, 33% to 90%, and 8% to 96%, respectively. The proportions of cows with FPR > 1.25, 1.35, 1.42, 1.50, 1.60, and 1.70 were 60%, 50%, 44%, 30%, 14%, and 6%, respectively. Incidences of clinical ketosis and milk yield were similar between cows that received 400 mL of PPG (n = 34) and control cows (n = 38). Prevalence of SCK at enrollment was 29.2%; therefore, FPR > 1.5 is not indicated for treatment. Lower cut-offs should be used for screening.

  7. THE CONSUMPTION OF RED PUPUNHA (BACTRIS GASIPAES KUNTH) INCREASES HDL CHOLESTEROL AND REDUCES WEIGHT GAIN OF LACTATING AND POST-LACTATING WISTAR RATS

    PubMed Central

    Carvalho, R. Piccolotto; Lemos, J.R. Gonzaga; de Aquino Sales, R. Souza; Martins, M. Gassen; Nascimento, C.H.; Bayona, M.; Marcon, J.L.; Monteiro, J. Barros

    2014-01-01

    Introduction The lactating and post-lactating periods are marked by large metabolic change. Production of milk is 60% lipid dependent. We reported in a recent scientific meeting that Red pupunha palm tree fruit increases HDL cholesterol in lactating rats. This study evaluated if consumption of Red Pupunha by adult female rats has a beneficial impact on the lipid metabolism of lacting and post-lacting adult rats. Objective Evaluate if consumption of red pupunha has a beneficial effect in the lipid metabolism of lacting and post-lacting adult Wistar rats. Research Methods Four groups including two for control; (1) control adult lactating rats, (2) control adults post-lactating rats; and two experimental groups; (3) pupunha adults lactating rats and (4) pupunha adult post-lactating rats were evaluated and compared regarding: weight gain, food consumption, plasma total protein, glucose, total lipid, triglycerides, total cholesterol and HDL-cholesterol levels. The mean difference and its 95% confidence intervals were used for group comparisons. Group comparisons were evaluated by using analysis of variance (one-way ANOVA). The statistical significance of the pairwise differences among groups was assessed by using the two-sided Tukey test. Results There were no important differences in food consumption, plasma glucose, total lipids and triglycerides among groups. The red pupunha lactating group gain less weight showing lower body mass index (BMI) than controls (p < 0.05). Total cholesterol was lower in red pupunha lactating than in controls but not in the red pupunha post-lactating group as compared to controls. Triglycerides were lower in the post-lactating red pupunha group as compared to the control group (p = 0.039) but not for the lactating groups. Red pupunha lactating and post-lactating groups had higher HDL-cholesterol than their corresponding control groups (p ≤ 0.01). Conclusion Original findings include the beneficial effect of red pupunha in post-lactating

  8. Lactate and lactate clearance in acute cardiac care patients

    PubMed Central

    Lazzeri, Chiara; Picariello, Claudio; Dini, Carlotta Sorini; Gensini, Gian Franco; Valente, Serafina

    2012-01-01

    Hyperlactataemia is commonly used as a diagnostic and prognostic tool in intensive care settings. Recent studies documented that serial lactate measurements over time (or lactate clearance), may be clinically more reliable than lactate absolute value for risk stratification in different pathological conditions. While the negative prognostic role of hyperlactataemia in several critical ill diseases (such as sepsis and trauma) is well established, data in patients with acute cardiac conditions (i.e. acute coronary syndromes) are scarce and controversial. The present paper provides an overview of the current available evidence on the clinical role of lactic acid levels and lactate clearance in acute cardiac settings (acute coronary syndromes, cardiogenic shock, cardiac surgery), focusing on its prognostic role. PMID:24062898

  9. Pelleted beet pulp substituted for high-moisture corn: 3. Effects on ruminal fermentation, pH, and microbial protein efficiency in lactating dairy cows.

    PubMed

    Voelker, J A; Allen, M S

    2003-11-01

    The effects of increasing concentrations of dried, pelleted beet pulp substituted for high-moisture corn on ruminal fermentation, pH, and microbial efficiency were evaluated using eight ruminally and duodenally cannulated multiparous Holstein cows in a duplicated 4 x 4 Latin square design with 21-d periods. Cows were 79 +/- 17 (mean +/- SD) DIM at the beginning of the experiment. Experimental diets with 40% forage (corn silage and alfalfa silage) and 60% concentrate contained 0, 6.1, 12.1, or 24.3% beet pulp substituted for high-moisture corn on a DM basis. Diet concentrations of NDF and starch were 24.3 and 34.6% (0% beet pulp), 26.2 and 30.5% (6% beet pulp), 28.0, and 26.5% (12% beet pulp), and 31.6 and 18.4% (24% beet pulp), respectively. Substituting beet pulp for corn did not affect daily mean or minimum ruminal pH but tended to reduce pH range. Ruminal acetate:propionate responded in a positive exponential relationship to added beet pulp. Rate of valerate absorption from the rumen was not affected by treatment. Substituting beet pulp for corn up to 24% of diet DM did not affect efficiency of ruminal microbial protein production, expressed as microbial N flow to the duodenum as a percentage of OM truly digested in the rumen. Microbial efficiency was not correlated to mean pH or daily minimum pH. While microbial efficiency was not directly related to concentration of beet pulp fed, it was positively correlated with passage rate of particulate matter, as represented by starch and indigestible NDF, probably due to reduced turnover of microbial protein in the rumen.

  10. The Many Axes of Deer Lactation.

    PubMed

    Ceacero, Francisco; García, Andrés J; Landete-Castillejos, Tomás; Komárková, Martina; Hidalgo, Francisco; Serrano, Martina P; Gallego, Laureano

    2016-12-01

    In undomesticated animals information about the production and composition of milk over time is still scarce. In general, for most mammals it is known that milk composition changes across lactation, is different for male and female offspring, and even that marsupials, such as kangaroos, can simultaneously produce milk of different compositions for young of different ages. Such parallel milk production of differing compositions has not yet been studied in single-offspring placental mammals, but may help to explain behavioural processes like allosuckling (feeding the young of other adults) and lateralized suckling preferences. In this study we analysed the production and composition of milk in red deer throughout the lactation period and now confirm for the first time that there are axial differences present. The front teats, which are the favoured suckling positions of the deer's offspring, produce milk with a greater protein-to-fat ratio. Also, from the beginning of lactation the yield is greater on the left side, the side preferred by calves in all of the studied species, both at population and individual level. The links between milk production and calf behaviour in deer deserve further study.

  11. Effects of Pregnancy and Lactation on Iron Metabolism in Rats

    PubMed Central

    Gao, Guofen; Liu, Shang-Yuan; Wang, Hui-Jie; Zhang, Tian-Wei; Yu, Peng; Duan, Xiang-Lin; Zhao, Shu-E; Chang, Yan-Zhong

    2015-01-01

    In female, inadequate iron supply is a highly prevalent problem that often leads to iron-deficiency anemia. This study aimed to understand the effects of pregnancy and lactation on iron metabolism. Rats with different days of gestation and lactation were used to determine the variations in iron stores and serum iron level and the changes in expression of iron metabolism-related proteins, including ferritin, ferroportin 1 (FPN1), ceruloplasmin (Cp), divalent metal transporter 1 (DMT1), transferrin receptor 1 (TfR1), and the major iron-regulatory molecule—hepcidin. We found that iron stores decline dramatically at late-pregnancy period, and the low iron store status persists throughout the lactation period. The significantly increased FPN1 level in small intestine facilitates digestive iron absorption, which maintains the serum iron concentration at a near-normal level to meet the increase of iron requirements. Moreover, a significant decrease of hepcidin expression is observed during late-pregnancy and early-lactation stages, suggesting the important regulatory role that hepcidin plays in iron metabolism during pregnancy and lactation. These results are fundamental to the understanding of iron homeostasis during pregnancy and lactation and may provide experimental bases for future studies to identify key molecules expressed during these special periods that regulate the expression of hepcidin, to eventually improve the iron-deficiency status. PMID:26788496

  12. Feed intake and production parameters of lactating crossbred cows fed maize-based diets of stover, silage or quality protein silage.

    PubMed

    Gebrehawariat, Efrem; Tamir, Berhan; Tegegne, Azage

    2010-12-01

    Thirty-six Boran × Friesian dairy cows (392 ± 12 kg; mean ± SD) in early parity were used in a randomised complete block design. Cows were blocked by parity into three blocks of 12 animals and offered normal maize (NM) stover (T1), NM silage (T2) or quality protein maize (QPM) silage (T3) basal diets supplemented with a similar concentrate mix. Feed intake, body weight and condition changes and milk yield and composition were assessed. The daily intake of DM, OM, NDF and ADF for cows fed the NM stover-based diet was higher (P<0.05) than for the cows fed the NM silage and QPM silage-based diets. However, the daily intake of DOM (9.3 kg) and ME (140.8 MJ) for cows on QPM silage-based diet was higher (P<0.05) than for cows on NM stover-based diet (8.4 kg and 124.2 MJ) and NM silage-based diet (7.9 kg and 119.1 MJ). Body weight of cows was affected (P<0.05) by the diet, but diet had no effect (P>0.05) on body condition score, milk yield and milk composition. The digestible organic matter in the NM stover-based diet (724 g/kg DM) was lower (P<0.05) than that in the NM (770 g/kg DM) and QPM silage-based diet (762 g/kg DM). It was concluded that the performances of the cows on the NM silage and QPM silage diets were similar and were not superior to that of the NM stover-based diet.

  13. Proteomic analysis of microsomes from lactating bovine mammary gland.

    PubMed

    Peng, Lifeng; Rawson, Pisana; McLauchlan, Danyl; Lehnert, Klaus; Snell, Russell; Jordan, T William

    2008-04-01

    Mammary gland has multiple metabolic potential including for large-scale synthesis of milk proteins, carbohydrate, and lipids including nutrient triacylglycerols. We have carried out a proteomic analysis of mammary tissue to discover proteins that affect lipid metabolism. Unfractionated microsomes from lactating bovine mammary tissue were analyzed using one-dimensional SDS-PAGE with RPLC-ESI-MS/MS. This approach gave 703 proteins including 160 predicted transmembrane proteins. Proteins were classified according to their subcellular localizations and biological functions. Over 50 proteins were associated with cellular uptake, metabolism, and secretion of lipids, including some enzymes that have been previously associated with breast cancer and potential therapeutic targets. This database develops a proteomic view of the metabolic potential of mammary gland that can be expected to contribute to a greater understanding of gene expression and tissue remodeling associated with lactation, and to further dissection of normal and pathological processes in mammary tissue.

  14. 21 CFR 73.165 - Ferrous lactate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 1 2014-04-01 2014-04-01 false Ferrous lactate. 73.165 Section 73.165 Food and... ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.165 Ferrous lactate. (a) Identity. The color additive ferrous lactate is the ferrous lactate defined in § 184.1311 of this chapter. (b) Specifications. Ferrous...

  15. 21 CFR 73.165 - Ferrous lactate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 1 2012-04-01 2012-04-01 false Ferrous lactate. 73.165 Section 73.165 Food and... ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.165 Ferrous lactate. (a) Identity. The color additive ferrous lactate is the ferrous lactate defined in § 184.1311 of this chapter. (b) Specifications. Ferrous...

  16. 21 CFR 73.165 - Ferrous lactate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 1 2013-04-01 2013-04-01 false Ferrous lactate. 73.165 Section 73.165 Food and... ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.165 Ferrous lactate. (a) Identity. The color additive ferrous lactate is the ferrous lactate defined in § 184.1311 of this chapter. (b) Specifications. Ferrous...

  17. 21 CFR 73.165 - Ferrous lactate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 1 2010-04-01 2010-04-01 false Ferrous lactate. 73.165 Section 73.165 Food and... ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.165 Ferrous lactate. (a) Identity. The color additive ferrous lactate is the ferrous lactate defined in § 184.1311 of this chapter. (b) Specifications. Ferrous...

  18. 21 CFR 73.165 - Ferrous lactate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 1 2011-04-01 2011-04-01 false Ferrous lactate. 73.165 Section 73.165 Food and... ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.165 Ferrous lactate. (a) Identity. The color additive ferrous lactate is the ferrous lactate defined in § 184.1311 of this chapter. (b) Specifications. Ferrous...

  19. Lactating Adenoma of the Breast.

    PubMed

    Barco Nebreda, Israel; Vidal, M Carmen; Fraile, Manel; Canales, Lydia; González, Clarisa; Giménez, Nuria; García-Fernández, Antonio

    2016-08-01

    Lactating adenoma is an uncommon breast palpable lesion occurring in pregnancy or lactation. Although it is a benign condition, it often requires core biopsy or even surgery to exclude malignancy. As with other solid lesions in pregnancy and lactation, lactating adenoma needs an accurate evaluation in order to ensure its benign nature. Work-up must include both imaging and histologic findings. Ultrasound evaluation remains the first step in assessing the features of the lesion. Some authors consider magnetic resonance imaging as a useful tool in cases of inconclusive evaluation after ultrasound and histologic exam in an attempt to avoid surgery. Most lactating adenomas resolve spontaneously, whereas others persist or even increase in size and must be removed. The authors present a case of a 35-year-old woman at 6 months postpartum with a lactating adenoma in her right breast. After surgical removal, breastfeeding was perfectly continued within the next 24 hours, which highlights the fact that breast surgery is most often compatible with breastfeeding. © The Author(s) 2016.

  20. Elevated parathyroid hormone-related peptide levels after human gestation: relationship to changes in bone and mineral metabolism.

    PubMed

    Dobnig, H; Kainer, F; Stepan, V; Winter, R; Lipp, R; Schaffer, M; Kahr, A; Nocnik, S; Patterer, G; Leb, G

    1995-12-01

    PTH-related peptide (PTHrP) can be found in high concentrations in human breast milk and has been implicated in material calcium regulation postpartum. We studied the relationship of plasma PTHrP levels of serum markers of bone turnover and selective cancellous bone density in 35 women (age, 25 +/- 3 yr) 2-3 days postpartum and after 3 and 6 months of lactation. The mean postpartum plasma PTHrP levels measured by immunoradiometric assay was 2.64 +/- 0.19 pmol/L (mean +/- SE) and were elevated compared to that in 35 age- and sex-matched controls (1.34 +/- 0.14; P < 0.0001). PTHrP remained significantly elevated, but decreased during the lactation period of 6 +/- 1 months. Immediately postpartum, serum protein levels were lowest, and serum ionized calcium levels highest. At that time, PTH was suppressed to 50% of control values (P < 0.001). Two or 3 days postpartum, serum ionized calcium was negatively correlated with total serum protein (r = -0.47; P < 0.0001) and positively correlated with plasma PTHrP (r = -0.32; P < 0.008). PTH was inversely correlated with ionized calcium (r = -0.24; P = 0.03) and PTHrP (r = -0.31; P < 0.01). Three and 6 months postpartum, serum protein and PTH levels had returned to normal, and ionized calcium concentrations decreased. There was no indication that PTHrP may have any significant systemic effect after 3 and 6 months of lactation. Long term lactation led to a significant decrease in radial cancellous bone density (-4.5%; P < 0.05) at 6 months and to elevations in serum markers of bone resorption (2- to 3-fold for serum carboxy-terminal telopeptide of type I collagen) and formation (1- to 2-fold for osteocalcin and serum carboxy-terminal propeptide of type I procollagen). Bone turnover balance was clearly negative after 3 months of lactation compared to the control value and indicated net bone loss at a time when estrogen levels were low. With ongoing lactation, estrogen levels increased, and bone turnover balance improved

  1. Replacing dietary soybean meal with canola meal improves production and efficiency of lactating dairy cows

    USDA-ARS?s Scientific Manuscript database

    Previous research suggested that crude protein (CP) from canola meal (CM) is used more efficiently than CP from solvent soybean meal (SBM) by lactating dairy cows. We tested whether dietary CP content influenced relative effectiveness of equal supplemental CP from either CM or SBM. Fifty lactating H...

  2. The effect of naringin on early growth and development of the spheno-occipital synchondrosis as measured by the expression of PTHrP and Sox9--an in vitro model.

    PubMed

    Nowak-Solinska, Eva; Rabie, A Bakr M; Wong, Ricky W K; Lei, Samuel W Y

    2013-12-01

    The aim of this study was to assess the effect of the flavonoid naringin on the growth of the spheno-occipital synchondrosis by quantifying the levels of expression of Sox9 and PTHrP in an in vitro mouse model. Fifty 1-day-old BALB/c mice were randomly assigned to experimental or control groups, and each group equally divided into five time frames (6, 24, 48, 72 and 168 hours). The mice were sacrificed with phenobarbitone sodium, and the spheno-occipital synchondroses dissected and cultured in control or experimental medium, with the experimental medium supplemented with 0.1 µm naringin. Sections of the specimens underwent immunohistochemical staining for Sox9 and PTHrP, and the amount of expression was quantified using true-colour RGB (red-green-blue) computer-assisted image-analysing system with digital imaging. Data analysis showed there was a significant increase of expression of Sox9 at 6 and 24 hours (P < 0.001) between experimental and control groups, however, there was no significant difference between the levels of expression of PTHrP between experimental and control groups at any of the time frames. There was a very weak correlation found in this study between the expression of PTHrP and Sox9. In conclusion, naringin enhances the growth of the spheno-occipital synchondrosis through over expression of Sox9. This is a successful in vitro model to study factors regulating the growth of the spheno-occipital synchondrosis.

  3. High-affinity HLA-A(*)02.01 peptides from parathyroid hormone-related protein generate in vitro and in vivo antitumor CTL response without autoimmune side effects.

    PubMed

    Francini, Guido; Scardino, Antonio; Kosmatopoulos, Kostas; Lemonnier, François A; Campoccia, Giuseppe; Sabatino, Marianna; Pozzessere, Daniele; Petrioli, Roberto; Lozzi, Luisa; Neri, Paolo; Fanetti, Giuseppe; Cusi, Maria Grazia; Correale, Pierpaolo

    2002-11-01

    Parathyroid hormone-related protein (PTH-rP), a protein produced by prostate carcinoma and other epithelial cancers, is a key agent in the development of bone metastases. We investigated whether the protein follows the self-tolerance paradigm or can be used as a target Ag for anticancer immunotherapy by investigating the immunogenicity of two HLA-A(*)02.01-binding PTH-rP-derived peptides (PTR-2 and -4) with different affinity qualities. PTH-rP peptide-specific CTL lines were generated from the PBMC of two HLA-A(*)02.01(+) healthy individuals, stimulated in vitro with PTH-rP peptide-loaded autologous dendritic cells and IL-2. The peptide-specific CTLs were able to kill PTH-rP(+)HLA-A(*)02.01(+) breast and prostate carcinoma cell lines. The two peptides were also able to elicit a strong antitumor PTH-rP-specific CTL response in HLA-A(*)02.01 (HHD) transgenic mice. The vaccinated mice did not show any sign of side effects due to cell-mediated autoimmunity or toxicity. In this study we describe two immunogenic and toxic-free PTH-rP peptides as valid candidates for the design of peptide-based vaccination strategies against prostate cancer and bone metastases from the most common epithelial malignancies.

  4. Novel Lactate Transporters from Carboxylic Acid-Producing Rhizopus

    USDA-ARS?s Scientific Manuscript database

    The fungus Rhizopus is frequently used for fermentative production of lactic acid, but little is known about the mechanisms or proteins for transporting this carboxylic acid. Since transport of the lactate anion across the plasma membrane is critical to prevent acidification of the cytoplasm, we ev...

  5. Analysis of a Functional Lactate Permease in the Fungus Rhizopus

    USDA-ARS?s Scientific Manuscript database

    The fungus Rhizopus is frequently used for fermentative production of lactic acid, but little is known about the mechanisms or proteins for transporting this carboxylic acid. Since transport of the lactate anion across the plasma membrane is critical to prevent acidification of the cytoplasm, we ev...

  6. Changes in mRNA expression of arcuate nucleus appetite-regulating peptides during lactation in rats.

    PubMed

    Suzuki, Yoshihiro; Nakahara, Keiko; Maruyama, Keisuke; Okame, Rieko; Ensho, Takuya; Inoue, Yoshiyuki; Murakami, Noboru

    2014-04-01

    The contribution of hypothalamic appetite-regulating peptides to further hyperphagia accompanying the course of lactation in rats was investigated by using PCR array and real-time PCR. Furthermore, changes in the mRNA expression for appetite-regulating peptides in the hypothalamic arcuate nucleus (ARC) were analyzed at all stages of pregnancy and lactation, and also after weaning. Food intake was significantly higher during pregnancy, lactation, and after weaning than during non-lactation periods. During lactation, ARC expression of mRNAs for agouti-related protein (AgRP) and peptide YY was increased, whereas that of mRNAs for proopiomelanocortin (POMC) and cholecystokinin (CCK) was decreased, in comparison with non-lactation periods. The increase in AgRP mRNA expression during lactation was especially marked. The plasma level of leptin was significantly decreased during the course of lactation, whereas that of acyl-ghrelin was unchanged. In addition, food intake was negatively correlated with the plasma leptin level during lactation. This study has clarified synchronous changes in the expression of many appetite-regulating peptides in ARC of rats during lactation. Our results suggest that hyperphagia during lactation in rats is caused by decreases in POMC and CCK expression and increases in AgRP expression in ARC, the latter being most notable. Together with the decrease in the blood leptin level, such changes in mRNA expression may explain the further hyperphagia accompanying the course of lactation.

  7. Nutritional aspects of human lactation*

    PubMed Central

    Thomson, A. M.; Black, A. E.

    1975-01-01

    This paper reviews the literature on the incidence and duration of breast-feeding in various countries, the volume and composition of breast milk, the health and nutrition of breast-fed babies as judged by growth and morbidity, maternal nutritional requirements during lactation, and the effect of prolonged lactation on maternal health. It appears that lactation can be as well sustained by impoverished as by affluent mothers, and that even in communities where malnutrition is common the average growth of infants is satisfactory up to the age of about 3 months on a diet of breast milk alone. Breast milk appears to have specific anti-infective properties, but prolonged breast-feeding will not prevent infections among older infants reared in a poor environment. The authors believe that breast-feeding is the best form of nutrition for the young infant and deplore its decline in modern industrial societies. The recommendations of various FAO/WHO Expert Groups on nutritional intakes during lactation are summarized. The need for an increased daily energy intake of 4.2 MJ (1 000 kcal) is questioned, and an increase of 2.5 MJ (600 kcal) is suggested. Data on the effect of prolonged lactation on the health of the mother are scanty; body weight appears to be maintained even among poorly nourished mothers. The authors stress the need for well-planned and technically adequate studies of the material and psychological factors involved in breast feeding. PMID:816479

  8. Hematological and biochemical findings in pregnant, postfoaling, and lactating jennies.

    PubMed

    Bonelli, F; Rota, A; Corazza, M; Serio, D; Sgorbini, M

    2016-04-15

    The aims of this study were to (1) verify if significant changes occur in hematological and biochemical parameters in jennies during the last 2 months of pregnancy and the first 2 months of lactation, and (2) determine any differences with mares. Hematological and biochemical parameters were evaluated in jennies every 15 days during late pregnancy, parturition, and early lactation. The Kolmogorov-Smirnov test, analysis of variance for repeated measurements and Tukey's multiple comparison test as post hoc were applied. The significance level was set at P < 0.05. Statistical analysis showed differences related to time for Red Blood Cells (RBC) count and Hematocrit (HCT), White Blood Cells (WBC) count, platelet count (PLT), total proteins, blood urea, triglycerides and total cholesterol concentrations, aspartate aminotransferase, gamma-glutamyltransferase, creatine-phosphokinase activities, sodium (Na) and potassium (K). RBC and HCT were higher in late pregnancy than at foaling and during lactation. The relative anemia might be due to increased water ingestion because of fluid losses. The WBC count was higher at foaling than during late pregnancy and lactation. This could be related to the release of cortisol and catecholamine during delivery. The PLT trend showed lower values from delivery to the first 2 months of lactation compared to late gestation. Blood urea increased near parturition, and then remained constant during delivery and lactation, which might be due to the high energy demand at the beginning of lactation. Triglycerides and total cholesterol showed a decrease from delivery through the lactation period. Thus, jennies seem to have a similar metabolism of fats to ponies and draft horse mares, characterized by a greater fat content and mobilization than light breed horses. Aspartate aminotransferase activity decreased at parturition and early lactation, probably because of a predominance of anabolic over catabolic processes during pregnancy. Gamma

  9. Prostate cancer cells metabolize d-lactate inside mitochondria via a D-lactate dehydrogenase which is more active and highly expressed than in normal cells.

    PubMed

    de Bari, Lidia; Moro, Loredana; Passarella, Salvatore

    2013-03-01

    Although D-lactate metabolism has been shown to occur in a variety of mitochondria, the metabolic fate of D-lactate in cancer cells has never been investigated, as it is believed to be exported to the extracellular phase. We show that mitochondria from both cancer (PC-3) and normal (PNT1A) prostate cells can metabolize D-lactate in an energy competent manner. This is due to the mitochondrial D-lactate dehydrogenase, a membrane flavoprotein, the activity and protein level of which are higher in PC-3 than in PNT1A cells, as detected by both kinetic and immunological analysis. D-Lactate can enter prostate mitochondria and cause the export of newly synthesized malate in a carrier-mediated manner, with the rate of malate efflux from mitochondria twofold higher in cancer. Copyright © 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  10. Comparative Proteomic Insights into the Lactate Responses of Halophilic Salinicoccus roseus W12

    PubMed Central

    Wang, Hongyan; Wang, Limin; Yang, Han; Cai, Yumeng; Sun, Lifan; Xue, Yanfen; Yu, Bo; Ma, Yanhe

    2015-01-01

    Extremophiles use adaptive mechanisms to survive in extreme environments, which is of great importance for several biotechnological applications. A halophilic strain, Salinicoccus roseus W12, was isolated from salt lake in Inner Mongolia, China in this study. The ability of the strain to survive under high sodium conditions (including 20% sodium lactate or 25% sodium chloride, [w/v]) made it an ideal host to screen for key factors related to sodium lactate resistance. The proteomic responses to lactate were studied using W12 cells cultivated with or without lactate stress. A total of 1,656 protein spots in sodium lactate-treated culture and 1,843 spots in NaCl-treated culture were detected by 2-dimensional gel electrophoresis, and 32 of 120 significantly altered protein spots (fold change > 2, p < 0.05) were identified by matrix-assisted laser-desorption ionization time-of-flight mass spectrometry. Among 21 successfully identified spots, 19 proteins were upregulated and 2 were downregulated. The identified proteins are mainly involved in metabolism, cellular processes and signaling, and information storage and processing. Transcription studies confirmed that most of the encoding genes were upregulated after the cells were exposed to lactate in 10 min. Cross-protecting and energy metabolism-related proteins played an important role in lactate tolerance for S. roseus W12. PMID:26358621

  11. Dietary ractopamine supplementation during the first lactation affects milk composition, piglet growth and sow reproductive performance.

    PubMed

    van Wettere, W H E J; Pain, S J; Hughes, P E

    2016-11-01

    Excessive mobilization of body reserves during lactation delays the return to reproductive function in weaned primiparous sows. This study tested the hypothesis that supplementing the lactation diets of first-parity sows with ractopamine hydrochloride would reduce maternal weight loss and improve subsequent reproductive performance. Gestating gilts were allocated to one of two treatment groups (n=30 sows/treatment), with one group fed a standard lactation diet (2.5g/Mcal LYS: DE) throughout lactation (CTRL), whereas the treatment group received the standard lactation diet supplemented with 10mg/kg ractopamine hydrochloride (RAC) from d 1 to 13 of lactation and 20mg/kg RAC from d 14 of lactation until artificial insemination (AI). Weaning occurred on d 21 of lactation, with AI occurring at the first post-weaning estrus. Compared to CTRL, RAC supplementation decreased (P<0.05) liveweight loss between d 13 and 20 of lactation (4.3±0.90 versus 1.3±0.96kg), and tended to increase (P=0.06) the number of second litter piglets born alive (9.5±0.52 versus 8.1±0.74). Treatment (RAC versus CTRL) reduced milk protein levels on d 13 and 20 of lactation (P<0.05), and piglet weight gain between d 13 and 20 of lactation (260±0.01 versus 310±0.01g/day, P<0.01). In conclusion, it is evident that dietary RAC altered milk composition and stimulated conservation of maternal body reserves during the third week of lactation, resulting in a beneficial effect on subsequent reproductive performance. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. A proton gradient is the driving force for uphill transport of lactate in human placental brush-border membrane vesicles.

    PubMed

    Balkovetz, D F; Leibach, F H; Mahesh, V B; Ganapathy, V

    1988-09-25

    The characteristics of lactate transport in brush-border membrane vesicles isolated from normal human full-term placentas were investigated. Lactate transport in these vesicles was Na+-independent, but was greatly stimulated when the extravesicular pH was made acidic. In the presence of an inwardly directed H+ gradient ([H+]o greater than [H+]i), transient uphill transport of lactate could be demonstrated. This H+ gradient-dependent stimulation was not a result of a H+ diffusion potential. Transport of lactate in the presence of the H+ gradient was not inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid or by furosemide, ruling out the participation of an anion exchanger in placental lactate transport. Many monocarboxylates strongly interacted with the lactate transport system, whereas, with the single exception of succinate, dicarboxylates did not. The monocarboxylates pyruvate and lactate, but not the dicarboxylate succinate, when present inside the vesicles, were able to exert a trans-stimulatory effect on the uptake of radiolabeled lactate. Kinetic analyses provided evidence for a single transport system with a Kt of 4.1 +/- 0.4 mM for lactate and a Vmax of 54.2 +/- 9.9 nmol/mg of protein/30 s. Pyruvate inhibited lactate transport competitively, by reducing the affinity of the system for lactate without altering the maximal velocity. It is concluded that human placental brush-border membranes possess a transport system specific for lactate and other monocarboxylates and that this transport system is Na+-independent and is energized by an inwardly directed H+ gradient. Lactate-H+ symport rather than lactate-OH- antiport appears to be the mechanism of the H+ gradient-dependent lactate transport in these membranes.

  13. Lactate Metabolism is Associated with Mammalian Mitochondria

    PubMed Central

    Chen, Ying-Jr; Mahieu, Nathaniel G.; Huang, Xiaojing; Singh, Manmilan; Crawford, Peter A; Johnson, Stephen L.; Gross, Richard W.; Schaefer, Jacob

    2016-01-01

    It is well established that lactate secreted by fermenting cells can be oxidized or used as a gluconeogenic substrate by other cells and tissues. Within the fermenting cell itself, however, it is generally assumed that lactate is produced to replenish NAD+ and then is secreted. Here we explored the possibility that cytosolic lactate is metabolized by the mitochondria of fermenting mammalian cells. We found that fermenting HeLa and H460 cells utilize exogenous lactate carbon to synthesize a large percentage of their lipids. With high-resolution mass spectrometry, we found that both 13C and 2-2H labels from enriched lactate enter the mitochondria. The lactate dehydrogenase (LDH) inhibitor oxamate decreased respiration of isolated mitochondria incubated in lactate, but not isolated mitochondria incubated in pyruvate. Additionally, transmission electron microscopy (TEM) showed that LDHB localizes to the mitochondria. Taken together, our results demonstrate a link between lactate metabolism and the mitochondria of fermenting mammalian cells. PMID:27618187

  14. Targeting lactate metabolism for cancer therapeutics.

    PubMed

    Doherty, Joanne R; Cleveland, John L

    2013-09-01

    Lactate, once considered a waste product of glycolysis, has emerged as a critical regulator of cancer development, maintenance, and metastasis. Indeed, tumor lactate levels correlate with increased metastasis, tumor recurrence, and poor outcome. Lactate mediates cancer cell intrinsic effects on metabolism and has additional non-tumor cell autonomous effects that drive tumorigenesis. Tumor cells can metabolize lactate as an energy source and shuttle lactate to neighboring cancer cells, adjacent stroma, and vascular endothelial cells, which induces metabolic reprogramming. Lactate also plays roles in promoting tumor inflammation and in functioning as a signaling molecule that stimulates tumor angiogenesis. Here we review the mechanisms of lactate production and transport and highlight emerging evidence indicating that targeting lactate metabolism is a promising approach for cancer therapeutics.

  15. Targeting lactate metabolism for cancer therapeutics

    PubMed Central

    Doherty, Joanne R.; Cleveland, John L.

    2013-01-01

    Lactate, once considered a waste product of glycolysis, has emerged as a critical regulator of cancer development, maintenance, and metastasis. Indeed, tumor lactate levels correlate with increased metastasis, tumor recurrence, and poor outcome. Lactate mediates cancer cell intrinsic effects on metabolism and has additional non–tumor cell autonomous effects that drive tumorigenesis. Tumor cells can metabolize lactate as an energy source and shuttle lactate to neighboring cancer cells, adjacent stroma, and vascular endothelial cells, which induces metabolic reprogramming. Lactate also plays roles in promoting tumor inflammation and in functioning as a signaling molecule that stimulates tumor angiogenesis. Here we review the mechanisms of lactate production and transport and highlight emerging evidence indicating that targeting lactate metabolism is a promising approach for cancer therapeutics. PMID:23999443

  16. Developmental regulation of mitochondrial biogenesis and function in the mouse mammary gland during a prolonged lactation cycle.

    PubMed

    Hadsell, Darryl L; Olea, Walter; Wei, Jerry; Fiorotto, Marta L; Matsunami, Risë K; Engler, David A; Collier, Robert J

    2011-03-29

    The regulation of mitochondrial biogenesis and function in the lactating mammary cell is poorly understood. The goal of this study was to use proteomics to relate temporal changes in mammary cell mitochondrial function during lactation to changes in the proteins that make up this organelle. The hypothesis tested was that changes in mammary cell mitochondrial biogenesis and function during lactation would be accounted for by coordinated changes in the proteins of the electron transport chain and that some of these proteins might be linked by their expression patterns to PPARGC1α and AMP kinase. The mitochondrial proteome was studied along with markers of mitochondrial biogenesis and function in mammary tissue collected from mice over the course of a single prolonged lactation cycle. Mammary tissue concentrations of AMP and ADP were increased (P < 0.05) during early lactation and then declined with prolonged lactation. Similar changes were also observed for mitochondrial ATP synthesis activity, mitochondrial mass and DNA copy number. Analysis of the mammary cell mitochondrial proteome identified 244 unique proteins. Of these, only two proteins of the electron transport chain were found to increase during early lactation. In contrast, coordinated changes in numerous electron transport chain proteins were observed both during mid- and late lactation. There were six proteins that could be directly linked to PPARGC1α through network analysis. Abundance of PPARGC-1α and phosphorylation of AMP kinase was highest on day 2 postpartum. The results suggest that the increases in mammary mitochondria ATP synthesis activity during early lactation results from changes in only a limited number proteins. In addition, decreases in a handful of proteins linked to lipid oxidation could be temporally linked to decreases in PPARGC1α and phospho-AMP kinase suggesting potential roles for these proteins in coordinating mammary gland metabolism during early lactation.

  17. Imaging Pregnant and Lactating Patients.

    PubMed

    Tirada, Nikki; Dreizin, David; Khati, Nadia J; Akin, Esma A; Zeman, Robert K

    2015-10-01

    As use of imaging in the evaluation of pregnant and lactating patients continues to increase, misperceptions of radiation and safety risks have proliferated, which has led to often unwarranted concerns among patients and clinicians. When radiologic examinations are appropriately used, the benefits derived from the information gained usually outweigh the risks. This review describes appropriateness and safety issues, estimated doses for imaging examinations that use iodizing radiation (ie, radiography, computed tomography, nuclear scintigraphy, and fluoroscopically guided interventional radiology), radiation risks to the mother and conceptus during various stages of pregnancy, and use of iodinated or gadolinium-based contrast agents and radiotracers in pregnant and lactating women. Maternal radiation risk must be weighed with the potential consequences of missing a life-threatening diagnosis such as pulmonary embolus. Fetal risks (ie, spontaneous abortion, teratogenesis, or carcinogenesis) vary with gestational age and imaging modality and should be considered in the context of the potential benefit of medically necessary diagnostic imaging. When feasible and medically indicated, modalities that do not use ionizing radiation (eg, magnetic resonance imaging) are preferred in pregnant and lactating patients. Radiologists should strive to minimize risks of radiation to the mother and fetus, counsel patients effectively, and promote a realistic understanding of risks related to imaging during pregnancy and lactation.

  18. 21 CFR 582.5311 - Ferrous lactate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Ferrous lactate. 582.5311 Section 582.5311 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5311 Ferrous lactate. (a) Product. Ferrous lactate. (b) Conditions of use. This substance...

  19. 21 CFR 582.1207 - Calcium lactate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Calcium lactate. 582.1207 Section 582.1207 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Additives § 582.1207 Calcium lactate. (a) Product. Calcium lactate. (b) Conditions of use. This substance...

  20. 21 CFR 582.1207 - Calcium lactate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Calcium lactate. 582.1207 Section 582.1207 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Additives § 582.1207 Calcium lactate. (a) Product. Calcium lactate. (b) Conditions of use. This substance...

  1. 21 CFR 582.1207 - Calcium lactate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Calcium lactate. 582.1207 Section 582.1207 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Additives § 582.1207 Calcium lactate. (a) Product. Calcium lactate. (b) Conditions of use. This substance...

  2. 21 CFR 582.1207 - Calcium lactate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Calcium lactate. 582.1207 Section 582.1207 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Additives § 582.1207 Calcium lactate. (a) Product. Calcium lactate. (b) Conditions of use. This substance...

  3. 21 CFR 582.1207 - Calcium lactate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Calcium lactate. 582.1207 Section 582.1207 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Additives § 582.1207 Calcium lactate. (a) Product. Calcium lactate. (b) Conditions of use. This substance...

  4. 21 CFR 582.5311 - Ferrous lactate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Ferrous lactate. 582.5311 Section 582.5311 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5311 Ferrous lactate. (a) Product. Ferrous lactate. (b) Conditions of use. This substance...

  5. 21 CFR 582.5311 - Ferrous lactate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Ferrous lactate. 582.5311 Section 582.5311 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5311 Ferrous lactate. (a) Product. Ferrous lactate. (b) Conditions of use. This substance...

  6. 21 CFR 582.5311 - Ferrous lactate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Ferrous lactate. 582.5311 Section 582.5311 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5311 Ferrous lactate. (a) Product. Ferrous lactate. (b) Conditions of use. This substance...

  7. Short communication: Early-lactation, but not mid-lactation, bovine lactoferrin preparation increases epithelial barrier integrity of Caco-2 cell layers.

    PubMed

    Anderson, Rachel C; Bassett, Shalome A; Haggarty, Neill W; Gopal, Pramod K; Armstrong, Kelly M; Roy, Nicole C

    2017-02-01

    Bovine lactoferrin is an important milk protein with many health-promoting properties, including improving intestinal barrier integrity. Dysfunction of this barrier, commonly referred to as "leaky gut," has been linked to inflammatory and autoimmune diseases. With some processing techniques, lactoferrin isolated from milk collected at the start of the milking season (early lactation) may have lower purity than that isolated from milk collected during the rest of the milking season (mid-lactation) and could result in differences in bioactivity based on the stage of lactation. We compared reversed-phase HPLC chromatographs of early-lactation and mid-lactation preparations and found that both had large chromatograph peaks at the time predicted for lactoferrin. The notable difference between the 2 chromatographs was a much larger peak in the early-lactation lactoferrin sample that was determined to be angiogenin. Angiogenin was first identified due to its ability to induce new blood vessel formation, but is now known to be involved in numerous physiological processes. Then, we compared the effects of early-lactation and mid-lactation lactoferrin preparations in 2 bioassays: trans-epithelial electrical resistance (TEER), a measure of intestinal barrier integrity, and peripheral blood mononuclear cell cytokine secretion, a measure of immune-stimulatory properties. We found that early-lactation lactoferrin increased TEER across Caco-2 cell layers compared with control from 10 to 48 h, mid-lactation lactoferrin did not alter TEER. We also found that early-lactation lactoferrin reduced the amount of IL-8 produced by peripheral blood mononuclear cells (compared with those treated with control medium) to a greater extent than mid-lactation lactoferrin. A pro-inflammatory chemokine, IL-8 is also known to decrease barrier function. These results suggest that the decrease in IL-8 production in the presence of early-lactation lactoferrin may be the mechanism by which it increases

  8. Establishment and characterization of a human ovarian small cell carcinoma, hypercalcemic type, cell line (OS-1) secreting PTH, PthrP and ACTH--special reference to the susceptibility of anti-cancer drugs.

    PubMed

    Ohi, Satoshi; Niimi, Shigeki; Okada, Naoya; Yamada, Kyosuke; Tachibana, Toshiaki; Hashimoto, Hisashi; Nakajima, Masako; Yasuda, Mitsuru; Tanaka, Tadao; Sato, Kahei; Ishikawa, Hiroshi

    2004-12-01

    We successfully established a novel cell line (OS-1) derived from human ovarian small cell carcinoma, hypercalcemic type secreted PTH, PTH-rP and ACTH. The OS-1 cell line was established from metastatic focus of uterus. A patient was 25-year-old Japanese woman. The first she received left ovariectomy on April 2002. The histopathological diagnosis was ovarian small cell carcinoma, pT2c, Nx, Mx. Then on June 2003, metastatic focus of uterus was ectomied. A part of the recurrent tumor of uterus was cut into small pieces with razor blades, and dissociated with 0.1% trypsin-0.02% EDTA/ PBS(-) solution at room temperature. The single cells and small cell clusters were seeded into 60mm dishes and cultured in growth medium (GM: DMEM/F12 supplemented with 20% fetal bovine serum and 0.1% non-essential amino acids solution) at 37 degrees C, 4.7% CO2 in humidified air. Medium was exchanged twice a week. The OS-1 cells grew as floating cultures in the dishes. Radioimmunoassay of the conditioned media was revealed that the cultures secreted large amount of PTH, PTHrP and ACTH simultaneously. Susceptibilities of anti-cancer drugs to the OS-1 cells were examined using oxygen electrode meter (Daikin), and the results suggested VLB and TXL were effective, and CDDP, CPT-11, VP-16, VCR, CPA, MMC and CBDCA were not effective. In our knowledge, it is the first report that the cell line secreting PTH, PTHrP and ACTH was successfully established from ovarian small cell carcinoma, hypercalcemic type. We expect that OS-1 cell line contribute to study on the mechanism of ectopic hormone secretion and susceptibility of anti cancer drugs to the small cell carcinoma.

  9. Evolution of D-lactate dehydrogenase activity from glycerol dehydrogenase and its utility for D-lactate production from lignocellulose

    PubMed Central

    Wang, Qingzhao; Ingram, Lonnie O.; Shanmugam, K. T.

    2011-01-01

    Lactic acid, an attractive, renewable chemical for production of biobased plastics (polylactic acid, PLA), is currently commercially produced from food-based sources of sugar. Pure optical isomers of lactate needed for PLA are typically produced by microbial fermentation of sugars at temperatures below 40 °C. Bacillus coagulans produces L(+)-lactate as a primary fermentation product and grows optimally at 50 °C and pH 5, conditions that are optimal for activity of commercial fungal cellulases. This strain was engineered to produce D(−)-lactate by deleting the native ldh (L-lactate dehydrogenase) and alsS (acetolactate synthase) genes to impede anaerobic growth, followed by growth-based selection to isolate suppressor mutants that restored growth. One of these, strain QZ19, produced about 90 g L-1 of optically pure D(−)-lactic acid from glucose in < 48 h. The new source of D-lactate dehydrogenase (D-LDH) activity was identified as a mutated form of glycerol dehydrogenase (GlyDH; D121N and F245S) that was produced at high levels as a result of a third mutation (insertion sequence). Although the native GlyDH had no detectable activity with pyruvate, the mutated GlyDH had a D-LDH specific activity of 0.8 μmoles min-1 (mg protein)-1. By using QZ19 for simultaneous saccharification and fermentation of cellulose to D-lactate (50 °C and pH 5.0), the cellulase usage could be reduced to 1/3 that required for equivalent fermentations by mesophilic lactic acid bacteria. Together, the native B. coagulans and the QZ19 derivative can be used to produce either L(+) or D(−) optical isomers of lactic acid (respectively) at high titers and yields from nonfood carbohydrates. PMID:22065761

  10. Evolution of D-lactate dehydrogenase activity from glycerol dehydrogenase and its utility for D-lactate production from lignocellulose.

    PubMed

    Wang, Qingzhao; Ingram, Lonnie O; Shanmugam, K T

    2011-11-22

    Lactic acid, an attractive, renewable chemical for production of biobased plastics (polylactic acid, PLA), is currently commercially produced from food-based sources of sugar. Pure optical isomers of lactate needed for PLA are typically produced by microbial fermentation of sugars at temperatures below 40 °C. Bacillus coagulans produces L(+)-lactate as a primary fermentation product and grows optimally at 50 °C and pH 5, conditions that are optimal for activity of commercial fungal cellulases. This strain was engineered to produce D(-)-lactate by deleting the native ldh (L-lactate dehydrogenase) and alsS (acetolactate synthase) genes to impede anaerobic growth, followed by growth-based selection to isolate suppressor mutants that restored growth. One of these, strain QZ19, produced about 90 g L(-1) of optically pure D(-)-lactic acid from glucose in < 48 h. The new source of D-lactate dehydrogenase (D-LDH) activity was identified as a mutated form of glycerol dehydrogenase (GlyDH; D121N and F245S) that was produced at high levels as a result of a third mutation (insertion sequence). Although the native GlyDH had no detectable activity with pyruvate, the mutated GlyDH had a D-LDH specific activity of 0.8 μmoles min(-1) (mg protein)(-1). By using QZ19 for simultaneous saccharification and fermentation of cellulose to D-lactate (50 °C and pH 5.0), the cellulase usage could be reduced to 1/3 that required for equivalent fermentations by mesophilic lactic acid bacteria. Together, the native B. coagulans and the QZ19 derivative can be used to produce either L(+) or D(-) optical isomers of lactic acid (respectively) at high titers and yields from nonfood carbohydrates.

  11. Reliability of urine lactate as a novel biomarker of lactate production capacity in maximal swimming.

    PubMed

    Nikolaidis, Stefanos; Karpouzi, Christina; Tsalis, George; Kabasakalis, Athanasios; Papaioannou, Konstantinos G; Mougios, Vassilis

    2016-01-01

    Postexercise urine lactate may be a novel biomarker of lactate production capacity during exercise. To evaluate the reliability and utility of the urine lactate concentration after maximal swimming trials between different training protocols (6 × 50 m and 3 × 100 m) and training states (active and nonactive swimmers). Lactate and creatinine were determined by spectrophotometry in blood and urine. Blood and urine lactate concentrations were correlated in-between training protocols and in participants of different training states. The reliability of the urine lactate concentration was moderate for one of the training protocols and good or moderate for the two training states. Additionally, it was lower than that of the blood lactate concentration, and did not improve after normalizing to the urine creatinine concentration. Although promising as a biomarker of lactate production capacity, urine lactate requires further research to improve its reliability.

  12. Lactate production under fully aerobic conditions: the lactate shuttle during rest and exercise.

    PubMed

    Brooks, G A

    1986-12-01

    O2 insufficiency and other factors increase the rate of lactate production. Significant quantities of lactate are produced under postabsorptive as well as postprandial conditions in resting individuals. In humans during postabsorptive rest, 25-50% of the total carbohydrate combusted appears to pass through the lactate pool. During sustained submaximal (in terms of VO2max) exercise, the rates of lactate production (Ri) and oxidation (Rox) are greatly elevated as compared to rest. However, lactate production and oxidation increase relatively less than O2 consumption during moderate-intensity exercise. Because the lactate production index (RiI = Ri/VO2) decreases during submaximal, moderate-intensity exercise compared to rest, it is concluded that skeletal muscle and other sites of lactate production are effectively oxygenated. Alterations in the levels of circulating catecholamines can affect levels and turnover rates of glucose and lactate. In pure red dog gracilis muscle in situ and in the healthy and myocardium in vivo, contraction results in glycolysis and lactate production. This production of lactate occurs despite an apparent abundance of O2. Similarly, glucose catabolism in the human brain results in lactate production. The formation of lactate under fully aerobic conditions of rest and exercise represents an important mechanism by which different tissues share a carbon source (lactate) for oxidation and other processes such as gluconeogenesis. This mechanism has been termed the lactate shuttle.

  13. Bladder cancer cells re-educate TAMs through lactate shuttling in the microfluidic cancer microenvironment.

    PubMed

    Zhao, Yang; Wang, Degui; Xu, Ting; Liu, Pengfei; Cao, Yanwei; Wang, Yonghua; Yang, Xuecheng; Xu, Xiaodong; Wang, Xinsheng; Niu, Haitao

    2015-11-17

    In the present study, we aimed to investigate the influence of lactate shuttling on the functional polarization and spatial distribution of transitional cell carcinoma of the bladder (TCCB) cells and macrophages. We designed a microfluidic coculture chip for real-time integrative assays. The effect of lactate shuttling on the re-education of macrophages by TCCB cells was explored by measuring the levels of NO using a total NO assay kit and by evaluating the protein expression of iNOS, p-NFkB-p65, Arg-1 and HIF-1α via cell immunofluorescence and western blotting. Additionally, we examined TCCB cell viability using acridine orange/ethidium bromide (AO/EB) and MitoTracker staining. Moreover, the concentration distributions of lactate and large signaling proteins in the culture chambers were measured using 4',6-diamidino-2-phenylindole (DAPI) and fluorescein isothiocyanate-dextran (FITC-dextran). Furthermore, the recruitment of macrophages and the influence of macrophages on BC metastasis were observed via light microscopy. We confirmed that TCCB cells reprogrammed macrophages into an M2 phenotype. Moreover, lactate inhibited M1 polarization and induced M2 polarization of macrophages, but blockade of cancer cell-macrophage lactate flux significantly inhibited the re-education of macrophages by TCCB cells. In addition, lactate diffused faster and deeper than large signaling proteins in the microfluidic tumor microenvironment. Furthermore, lactate alone induced the migration of macrophages, and M1, but not M2, macrophages reduced the motility of TCCB cells. TCCB cells reprogrammed macrophages into an M2 phenotype in a manner that depended on cancer cell-TAM lactate flux. Furthermore, the lactate shuttle may be a determinant of the density of TAMs in tumor tissue.

  14. Relationship among blood indicators of lipomobilization and hepatic function during early lactation in high-yielding dairy cows

    PubMed Central

    Muiño, Rodrigo; Pereira, Víctor; Campos, Rómulo; Benedito, José Luis

    2011-01-01

    Blood indicators are used as a tool to diagnose metabolic disorders. The present work was conducted to study the relationships among blood indicators of lipomobilization and hepatic function in high-yielding dairy cows. Two groups of Holstein cows were studied: 27 early lactation cows and 14 mid lactation cows from four different herds with similar husbandry characteristics in Galicia, Spain. Blood samples were obtained to measure beta-hydroxybutyrate (BHB), non-esterified fatty acids (NEFA), triglycerides (TG), and the activity of aspartate transaminase (AST) and gamma-glutamyl transferase. Cows in early lactation had higher levels of BHB and NEFA than mid lactation cows. High lipomobilization (NEFA > 400 µmol/L) was detected in 67% and 7% of early lactation and mid lactation cows, respectively, while subclinical ketosis (BHB > 1.2 mmol/L) was detected in 41% and 28% of the early lactation and lactation cows, respectively. TG concentrations were low in all cows suffering subclinical ketosis and in 61% of the cows with high lipomobilization. During early lactation, 30% of cows suffered hepatic lipidosis as detected by levels of AST. Compromised hepatic function was observed in early lactation cows as shown by lower concentrations of glucose, total protein, and urea. PMID:21897097

  15. Developmental regulation of mitochondrial biogenesis and function in the mouse mammary gland during a prolonged lactation cycle

    USDA-ARS?s Scientific Manuscript database

    The regulation of mitochondrial biogenesis and function in the lactating mammary cell is poorly understood. The goal of this study was to use proteomics to relate temporal changes in mammary cell mitochondrial function during lactation to changes in the proteins that make up this organelle. The hypo...

  16. Is lactate an undervalued functional component of fermented food products?

    PubMed Central

    Garrote, Graciela L.; Abraham, Analía G.; Rumbo, Martín

    2015-01-01

    Although it has been traditionally regarded as an intermediate of carbon metabolism and major component of fermented dairy products contributing to organoleptic and antimicrobial properties of food, there is evidence gathered in recent years that lactate has bioactive properties that may be responsible of broader properties of functional foods. Lactate can regulate critical functions of several key players of the immune system such as macrophages and dendritic cells, being able to modulate inflammatory activation of epithelial cells as well. Intraluminal levels of lactate derived from fermentative metabolism of lactobacilli have been shown to modulate inflammatory environment in intestinal mucosa. The molecular mechanisms responsible to these functions, including histone deacetylase dependent-modulation of gene expression and signaling through G-protein coupled receptors have started to be described. Since lactate is a major fermentation product of several bacterial families with probiotic properties, we here propose that it may contribute to some of the properties attributed to these microorganisms and in a larger view, to the properties of food products fermented by lactic acid bacteria. PMID:26150815

  17. Lactate Inhibits the Pro-Inflammatory Response and Metabolic Reprogramming in Murine Macrophages in a GPR81-Independent Manner

    PubMed Central

    Marchetti, Philippe; Tang, Cong; Kluza, Jerome; Offermanns, Stefan; Sirard, Jean-Claude; Rumbo, Martin

    2016-01-01

    Lactate is an essential component of carbon metabolism in mammals. Recently, lactate was shown to signal through the G protein coupled receptor 81 (GPR81) and to thus modulate inflammatory processes. This study demonstrates that lactate inhibits pro-inflammatory signaling in a GPR81-independent fashion. While lipopolysaccharide (LPS) triggered expression of IL-6 and IL-12 p40, and CD40 in bone marrow-derived macrophages, lactate was able to abrogate these responses in a dose dependent manner in Gpr81-/- cells as well as in wild type cells. Macrophage activation was impaired when glycolysis was blocked by chemical inhibitors. Remarkably, lactate was found to inhibit LPS-induced glycolysis in wild type as well as in Gpr81-/- cells. In conclusion, our study suggests that lactate can induce GPR81-independent metabolic changes that modulate macrophage pro-inflammatory activation. PMID:27846210

  18. Evolutionary well-conserved region in the signal peptide of parathyroid hormone-related protein is critical for its dual localization through the regulation of ER translocation

    PubMed Central

    Amaya, Yoshihiro; Nakai, Toshiki; Miura, Satoshi

    2016-01-01

    Parathyroid hormone-related protein (PTHrP) has two different targeting signals: an N-terminal signal peptide for the endoplasmic reticulum (ER) targeting and an internal nuclear localization signal. The protein not only functions as a secretory protein, but is also found in the nucleus and/or nucleolus under certain conditions. PTHrP signal peptide is less hydrophobic than most signal peptides mainly due to its evolutionarily well-conserved region (QQWS). The substitution of four tandem leucine residues for this conserved region resulted in a significant inhibition of the signal peptide cleavage. At the same time, proportion of nuclear and/or nucleolar localization decreased, probably due to tethering of the protein to the ER membrane by the uncleaved mutant signal peptide. Almost complete cleavage of the signal peptide accompanied by a lack of nuclear/nucleolar localization was achieved by combining the hydrophobic h-region and an optimized sequence of the cleavage site. In addition, mutational modifications of the distribution of charged residues in and around the signal peptide affect its cleavage and/or nuclear/nucleolar localization of the protein. These results indicate that the well-conserved region in the signal peptide plays an essential role in the dual localization of PTHrP through ER targeting and/or the membrane translocation. PMID:26538570

  19. Evolutionary well-conserved region in the signal peptide of parathyroid hormone-related protein is critical for its dual localization through the regulation of ER translocation.

    PubMed

    Amaya, Yoshihiro; Nakai, Toshiki; Miura, Satoshi

    2016-04-01

    Parathyroid hormone-related protein (PTHrP) has two different targeting signals: an N-terminal signal peptide for the endoplasmic reticulum (ER) targeting and an internal nuclear localization signal. The protein not only functions as a secretory protein, but is also found in the nucleus and/or nucleolus under certain conditions. PTHrP signal peptide is less hydrophobic than most signal peptides mainly due to its evolutionarily well-conserved region (QQWS). The substitution of four tandem leucine residues for this conserved region resulted in a significant inhibition of the signal peptide cleavage. At the same time, proportion of nuclear and/or nucleolar localization decreased, probably due to tethering of the protein to the ER membrane by the uncleaved mutant signal peptide. Almost complete cleavage of the signal peptide accompanied by a lack of nuclear/nucleolar localization was achieved by combining the hydrophobic h-region and an optimized sequence of the cleavage site. In addition, mutational modifications of the distribution of charged residues in and around the signal peptide affect its cleavage and/or nuclear/nucleolar localization of the protein. These results indicate that the well-conserved region in the signal peptide plays an essential role in the dual localization of PTHrP through ER targeting and/or the membrane translocation. © The Authors 2015. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

  20. Identification of epsilon-crystallin from swan lens as lactate dehydrogenase.

    PubMed

    Chiou, S H; Chang, W P; Lai, T A

    1989-10-01

    Characterization of lens crystallins from black swan, a rare aquatic bird belonging to the family Anatidae, was carried out to search for epsilon-crystallin with lactate dehydrogenase activity. Biochemical comparison of epsilon-crystallins isolated from the swan and duck lenses plus lactate dehydrogenase of chicken heart has also been made in order to establish the structural/functional relatedness of these proteins. Amino acid analyses showed essentially similar overall compositions for these three proteins. Kinetic analysis revealed differences between avian epsilon-crystallins and the authentic heart-type lactate dehydrogenase. The swan lenses similar to duck lenses appeared to contain a thermostable epsilon-crystallin which possesses very high enzymatic activity of lactate dehydrogenase. The characterization of epsilon-crystallins from the available species of aquatic birds may provide some insights into the evolution of this unique crystallin in the Aves and their enzymatic roles inside the lens.

  1. Identification of c-crystallin from swan lens as lactate dehydrogenase.

    PubMed

    Chiou, S-H; Chang, W-P; Lai, T-A

    1989-01-01

    Chuacterization of lens crystallins from black swan, a rare aquatic bird belonging to the family Anatidae, was carried out to search for e-crystallin with lactate dehydrogenase activity. Biochemical comparison of E-crystallins isolated from the swan and duck lenses plus lactate dehydrogenase of chicken heart has also been made in order to establish the structural/functional relatedness of these proteins. Amino acid analyses showed essentially similar overall compositions for these three proteins. Kinetic analysis revealed differences between avian E-crystallins and the authentic heart-type lactate dehydrogenase. The swan lenses similar to duck lenses appeared to contain a thermostable E-crystallin which possesses very high enzymatic activity of lactate dehydrogenase. The characterization of E-crystallins from the available species of aquatic birds may provide some insights into the evolution of this unique crystallin in the Aves and their enzymatic roles inside the lens.

  2. Exogenous lactate supply affects lactate kinetics of rainbow trout, not swimming performance

    PubMed Central

    Omlin, Teye; Langevin, Karolanne

    2014-01-01

    Intense swimming causes circulatory lactate accumulation in rainbow trout because lactate disposal (Rd) is not stimulated as strongly as lactate appearance (Ra). This mismatch suggests that maximal Rd is limited by tissue capacity to metabolize lactate. This study uses exogenous lactate to investigate what constrains maximal Rd and minimal Ra. Our goals were to determine how exogenous lactate affects: 1) Ra and Rd of lactate under baseline conditions or during graded swimming, and 2) exercise performance (critical swimming speed, Ucrit) and energetics (cost of transport, COT). Results show that exogenous lactate allows swimming trout to boost maximal Rd lactate by 40% and reach impressive rates of 56 μmol·kg−1·min−1. This shows that the metabolic capacity of tissues for lactate disposal is not responsible for setting the highest Rd normally observed after intense swimming. Baseline endogenous Ra (resting in normoxic water) is not significantly reduced by exogenous lactate supply. Therefore, trout have an obligatory need to produce lactate, either as a fuel for oxidative tissues and/or from organs relying on glycolysis. Exogenous lactate does not affect Ucrit or COT, probably because it acts as a substitute for glucose and lipids rather than extra fuel. We conclude that the observed 40% increase in Rd lactate is made possible by accelerating lactate entry into oxidative tissues via monocarboxylate transporters (MCTs). This observation together with the weak expression of MCTs and the phenomenon of white muscle lactate retention show that lactate metabolism of rainbow trout is significantly constrained by transmembrane transport. PMID:25121611

  3. Associations of insulin resistance later in lactation on fertility of dairy cows.

    PubMed

    Baruselli, P S; Vieira, L M; Sá Filho, M F; Mingoti, R D; Ferreira, R M; Chiaratti, M R; Oliveira, L H; Sales, J N; Sartori, R

    2016-07-01

    The challenge of getting dairy cows pregnant during early lactation is a well-described, worldwide problem. However, specifically in farms with poor reproductive, nutritional, and environmental conditions/management, a low pregnancy rate during early lactation is followed inevitably by an increased number of nonpregnant cows after 150 days in milk, with even more difficulties to achieve pregnancy. Therefore, several studies were designed to understand and develop strategies to mitigate reduced fertility of cows during late lactation. Experiments were performed under tropical regions to determine metabolic status during lactation and association of stage of lactation on oocyte quality and fertility. Lactating cows with extended days not pregnant (e.g.,>150 days in milk) often had systemic metabolic alterations, including development of peripheral insulin resistance and various oocyte alterations, including reduced expression of genes encoding glucose transport proteins, reduced amounts of mtDNA, increased expression of mitochondria-related genes, and increased expression of apoptosis-related genes. Additionally, in vitro embryo production and pregnancy per AI were lower in late- versus early-lactation cows in some but not all studies. Notwithstanding, when a normal embryo was transferred to a cow in late lactation, the pregnancy per transfer was reasonable, reinforcing the assertion that fertility problems in late-lactation cows may be associated with oocyte quality, fertilization, and/or failure of early embryo development. In conclusion, insulin resistance may reduce oocyte competence and consequently fertility in late-lactation dairy cows. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Oligodendrocyte Progenitor Cells Directly Utilize Lactate for Promoting Cell Cycling and Differentiation.

    PubMed

    Ichihara, Yoshinori; Doi, Toru; Ryu, Youngjae; Nagao, Motoshi; Sawada, Yasuhiro; Ogata, Toru

    2017-05-01

    Oligodendrocyte progenitor cells (OPCs) undergo marked morphological changes to become mature oligodendrocytes, but the metabolic resources for this process have not been fully elucidated. Although lactate, a metabolic derivative of glycogen, has been reported to be consumed in oligodendrocytes as a metabolite, and to ameliorate hypomyelination induced by low glucose conditions, it is not clear about the direct contribution of lactate to cell cycling and differentiation of OPCs, and the source of lactate for remyelination. Therefore, we evaluated the effect of 1,4-dideoxy-1,4-imino-d-arabinitol (DAB), an inhibitor of the glycogen catabolic enzyme glycogen phosphorylase, in a mouse cuprizone model. Cuprizone induced demyelination in the corpus callosum and remyelination occurred after cuprizone treatment ceased. This remyelination was inhibited by the administration of DAB. To further examine whether lactate affects proliferation or differentiation of OPCs, we cultured mouse primary OPC-rich cells and analyzed the effect of lactate. Lactate rescued the slowed cell cycling induced by 0.4 mM glucose, as assessed by the BrdU-positive cell ratio. Lactate also promoted OPC differentiation detected by monitoring the mature oligodendrocyte marker myelin basic protein, in the presence of both 36.6 mM and 0.4 mM glucose. Furthermore, these lactate-mediated effects were suppressed by the reported monocarboxylate transporter inhibitor, α-cyano-4-hydroxy-cinnamate. These results suggest that lactate directly promotes the cell cycling rate and differentiation of OPCs, and that glycogen, one of the sources of lactate, contributes to remyelination in vivo. J. Cell. Physiol. 232: 986-995, 2017. © 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc.

  5. Inflammatory mediators in mastitis and lactation insufficiency.

    PubMed

    Ingman, Wendy V; Glynn, Danielle J; Hutchinson, Mark R

    2014-07-01

    Mastitis is a common inflammatory disease during lactation that causes reduced milk supply. A growing body of evidence challenges the central role of pathogenic bacteria in mastitis, with disease severity associated with markers of inflammation rather than infection. Inflammation in the mammary gland may be triggered by microbe-associated molecular patterns (MAMPs) as well as danger-associated molecular patterns (DAMPs) binding to pattern recognition receptors such as the toll-like receptors (TLRs) on the surface of mammary epithelial cells and local immune cell populations. Activation of the TLR4 signalling pathway and downstream nuclear factor kappa B (NFkB) is critical to mediating local mammary gland inflammation and systemic immune responses in mouse models of mastitis. However, activation of NFkB also induces epithelial cell apoptosis and reduced milk protein synthesis, suggesting that inflammatory mediators activated during mastitis promote partial involution. Perturbed milk flow, maternal stress and genetic predisposition are significant risk factors for mastitis, and could lead to a heightened TLR4-mediated inflammatory response, resulting in increased susceptibility and severity of mastitis disease in the context of low MAMP abundance. Therefore, heightened host inflammatory signalling may act in concert with pathogenic or commensal bacterial species to cause both the inflammation associated with mastitis and lactation insufficiency. Here, we present an alternate paradigm to the widely held notion that breast inflammation is driven principally by infectious bacterial pathogens, and suggest there may be other therapeutic strategies, apart from the currently utilised antimicrobial agents, that could be employed to prevent and treat mastitis in women.

  6. Dietary intakes and food taboos of lactating women in a South Indian community.

    PubMed

    Sundararaj, R; Pereira, S M

    1975-06-01

    The diets of women, studied at regular intervals in the first year of lactation, showed that food taboos adversely affected the daily intake of energy, protein and some nutrients in the first month. Despite the poor intakes, the women breastfed their babies successfully and the weight gained by the babies in the early months was satisfactory. Breast feeding continued until the next pregnancy. Most women believed lactation prevented conception.

  7. 21 CFR 184.1639 - Potassium lactate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Potassium lactate. 184.1639 Section 184.1639 Food... Specific Substances Affirmed as GRAS § 184.1639 Potassium lactate. (a) Potassium lactate (C3H5O3K, CAS Reg. No. 996-31-6) is the potassium salt of lactic acid. It is a hydroscopic, white, odorless solid and is...

  8. 21 CFR 184.1639 - Potassium lactate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Potassium lactate. 184.1639 Section 184.1639 Food... Specific Substances Affirmed as GRAS § 184.1639 Potassium lactate. (a) Potassium lactate (C3H5O3K, CAS Reg. No. 996-31-6) is the potassium salt of lactic acid. It is a hydroscopic, white, odorless solid and is...

  9. 21 CFR 184.1639 - Potassium lactate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Potassium lactate. 184.1639 Section 184.1639 Food... GRAS § 184.1639 Potassium lactate. (a) Potassium lactate (C3H5O3K, CAS Reg. No. 996-31-6) is the potassium salt of lactic acid. It is a hydroscopic, white, odorless solid and is prepared commercially by...

  10. 21 CFR 184.1639 - Potassium lactate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Potassium lactate. 184.1639 Section 184.1639 Food... Specific Substances Affirmed as GRAS § 184.1639 Potassium lactate. (a) Potassium lactate (C3H5O3K, CAS Reg. No. 996-31-6) is the potassium salt of lactic acid. It is a hydroscopic, white, odorless solid and is...

  11. 21 CFR 184.1639 - Potassium lactate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Potassium lactate. 184.1639 Section 184.1639 Food... Specific Substances Affirmed as GRAS § 184.1639 Potassium lactate. (a) Potassium lactate (C3H5O3K, CAS Reg. No. 996-31-6) is the potassium salt of lactic acid. It is a hydroscopic, white, odorless solid and is...

  12. 21 CFR 184.1768 - Sodium lactate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Sodium lactate. 184.1768 Section 184.1768 Food and... Substances Affirmed as GRAS § 184.1768 Sodium lactate. (a) Sodium lactate (C3H5O3Na, CAS Reg. No. 72-17-3) is the sodium salt of lactic acid. It is prepared commercially by the neutralization of lactic acid...

  13. 21 CFR 184.1768 - Sodium lactate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Sodium lactate. 184.1768 Section 184.1768 Food and....1768 Sodium lactate. (a) Sodium lactate (C3H5O3Na, CAS Reg. No. 72-17-3) is the sodium salt of lactic acid. It is prepared commercially by the neutralization of lactic acid with sodium hydroxide. (b)...

  14. 21 CFR 184.1768 - Sodium lactate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium lactate. 184.1768 Section 184.1768 Food and... Substances Affirmed as GRAS § 184.1768 Sodium lactate. (a) Sodium lactate (C3H5O3Na, CAS Reg. No. 72-17-3) is the sodium salt of lactic acid. It is prepared commercially by the neutralization of lactic acid...

  15. 21 CFR 184.1768 - Sodium lactate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Sodium lactate. 184.1768 Section 184.1768 Food and... Substances Affirmed as GRAS § 184.1768 Sodium lactate. (a) Sodium lactate (C3H5O3Na, CAS Reg. No. 72-17-3) is the sodium salt of lactic acid. It is prepared commercially by the neutralization of lactic acid...

  16. 21 CFR 184.1768 - Sodium lactate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Sodium lactate. 184.1768 Section 184.1768 Food and... Substances Affirmed as GRAS § 184.1768 Sodium lactate. (a) Sodium lactate (C3H5O3Na, CAS Reg. No. 72-17-3) is the sodium salt of lactic acid. It is prepared commercially by the neutralization of lactic acid...

  17. 21 CFR 184.1207 - Calcium lactate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Calcium lactate. 184.1207 Section 184.1207 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR... Specific Substances Affirmed as GRAS § 184.1207 Calcium lactate. (a) Calcium lactate (C6H10CaO6.xH2O,...

  18. 21 CFR 184.1207 - Calcium lactate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Calcium lactate. 184.1207 Section 184.1207 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR... Specific Substances Affirmed as GRAS § 184.1207 Calcium lactate. (a) Calcium lactate (C6H10CaO6.xH2O,...

  19. 21 CFR 184.1207 - Calcium lactate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium lactate. 184.1207 Section 184.1207 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1207 Calcium lactate. (a) Calcium lactate (C6H10CaO6.xH2O, where x is...

  20. 21 CFR 184.1207 - Calcium lactate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Calcium lactate. 184.1207 Section 184.1207 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) DIRECT... GRAS § 184.1207 Calcium lactate. (a) Calcium lactate (C6H10CaO6.xH2O, where x is any integer up to...

  1. 21 CFR 184.1207 - Calcium lactate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Calcium lactate. 184.1207 Section 184.1207 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR... Specific Substances Affirmed as GRAS § 184.1207 Calcium lactate. (a) Calcium lactate (C6H10CaO6.xH2O,...

  2. Trans-generational effect of maternal lactation during pregnancy: a Holstein cow model.

    PubMed

    González-Recio, Oscar; Ugarte, Eva; Bach, Alex

    2012-01-01

    Epigenetic regulation in mammals begins in the first stages of embryogenesis. This prenatal programming determines, in part, phenotype expression in adult life. Some species, particularly dairy cattle, are conceived during the maternal lactation, which is a period of large energy and nutrient needs. Under these circumstances, embryo and fetal development compete for nutrients with the mammary gland, which may affect prenatal programming and predetermine phenotype at adulthood. Data from a specialized dairy breed were used to determine the transgenerational effect when embryo development coincides with maternal lactation. Longitudinal phenotypic data for milk yield (kg), ratio of fat-protein content in milk during first lactation, and lifespan (d) from 40,065 cows were adjusted for environmental and genetic effects using a Bayesian framework. Then, the effect of different maternal circumstances was determined on the residuals. The maternal-related circumstances were 1) presence of lactation, 2) maternal milk yield level, and 3) occurrence of mastitis during embryogenesis. Females born to mothers that were lactating while pregnant produced 52 kg (MonteCarlo standard error; MCs.e. = 0.009) less milk, lived 16 d (MCs.e. = 0.002) shorter and were metabolically less efficient (+0.42% milk fat/protein ratio; MCs.e.<0.001) than females whose fetal life developed in the absence of maternal lactation. The greater the maternal milk yield during embryogenesis, the larger the negative effects of prenatal programming, precluding the offspring born to the most productive cows to fully express their potential additive genetic merit during their adult life. Our data provide substantial evidence of transgenerational effect when pregnancy and lactation coincide. Although this effect is relatively low, it should not be ignored when formulating rations for lactating and pregnant cows. Furthermore, breeding, replacement, and management strategies should also take into account whether

  3. ABCG5/ABCG8-independent biliary cholesterol excretion in lactating rats

    PubMed Central

    Coy, Donna J.; Wooton-Kee, Clavia R.; Yan, Baoxiang; Sabeva, Nadezhda; Su, Kai; Graf, Gregory

    2010-01-01

    Lactation is associated with increased expression of bile acid transporters and an increased size and hydrophobicity of the bile acid pool in rats. ATP-binding cassette (ABC) transporters multidrug resistance protein 2 (Mdr2), Abcb11 [bile salt export pump (Bsep)], and Abcg5/Abcg8 heterodimers are essential for the biliary secretion of phospholipids, bile acids, and cholesterol, respectively. We investigated the expression of these transporters and secretion of their substrates in female control and lactating Sprague Dawley rats and C57BL/6 mice. Expression of Abcg5/Abcg8 mRNA was decreased by 97 and 60% by midlactation in rats and mice, respectively; protein levels of Abcg8 were below detection limits in lactating rats. Mdr2 mRNA expression was decreased in lactating rats and mice by 47 and 59%, respectively. Despite these changes in transporter expression, basal concentrations of cholesterol and phospholipid in bile were unchanged in rats and mice, whereas increased Bsep mRNA expression in early lactation coincided with an increased basal biliary bile acid concentration in lactating mice. Following taurocholate infusion, coupling of phospholipid and taurocholate secretion in bile of lactating mice was significantly impaired relative to control mice, with no significant changes in maximal secretion of cholesterol or bile acids. In rats, taurocholate infusion revealed a significantly impaired coupling of cholesterol to taurocholate secretion in bile in lactating vs. control animals. These data reveal marked utilization of an Abcg5/Abcg8-independent mechanism for basal biliary cholesterol secretion in rats during lactation, but a dependence on Abcg5/g8 for maximal biliary cholesterol secretion. PMID:20413720

  4. DIGE based proteome analysis of mammary gland tissue in water buffalo (Bubalus bubalis): lactating vis-a-vis heifer.

    PubMed

    Jena, Manoj K; Janjanam, Jagadeesh; Naru, Jasmine; Kumar, Saravanan; Kumar, Sudarshan; Singh, Surender; Mohapatra, Sushil K; Kola, Srujana; Anand, Vijay; Jaswal, Shalini; Verma, Arvind K; Malakar, Dhruba; Dang, Ajay K; Kaushik, Jai K; Reddy, Vanga S; Mohanty, Ashok K

    2015-04-24

    Mammary gland is an exocrine and sebaceous gland made up of branching network of ducts that end in alveoli. Milk is synthesized in the alveoli and secreted into alveolar lumen. Mammary gland represents an ideal system for the study of organogenesis that undergoes successive cycles of pregnancy, lactation and involution. To gain insights on the molecular events that take place in pubertal and lactating mammary gland, we have identified 43 differentially expressed proteins in mammary tissue of heifer (non-lactating representing a virgin mammary gland), and lactating buffaloes (Bubalus bubalis) by 2D-difference gel electrophoresis (2D-DIGE) and mass spectrometry. Twenty one proteins were upregulated during lactation whereas 8 proteins were upregulated in heifer mammary gland significantly (p<0.05). Bioinformatics analyses of the identified proteins showed that a majority of the proteins are involved in metabolic processes. The differentially expressed proteins were validated by real-time PCR and Western blotting. We observed differential expressions of certain new proteins including EEF1D, HSPA5, HSPD1 and PRDX6 during lactation which have not been reported before. The differentially expressed proteins were mapped to available biological pathways and networks involved in lactation. This study signifies the importance of some proteins which are preferentially expressed during lactation and in heifer mammary gland. This work is important because we have generated information in water buffalo (B. bubalis) for the first time which is the major milk producing animal in Indian Subcontinent. Out of a present production of 133milliontons of milk produced in India, contribution of buffalo milk is around 54%. Its physiology is somewhat different from the lactating cows. Buffalo milk composition varies from cow milk in terms of higher fat and total solid content, which confers an advantage in preparation of specialized cheese, curd and other dairy products. Being a major milk

  5. Induced lactation: gaining a better understanding.

    PubMed

    Wittig, Sarah L; Spatz, Diane L

    2008-01-01

    Although a small minority in the United States, a number of adoptive mothers continue to seek information regarding the induction of lactation. Because of the level of support needed by these women to successfully induce lactation, it is necessary for nurses and other healthcare workers to gain a thorough understanding of the various processes and medications frequently used. Often, women who induce lactation cannot produce enough milk to exclusively breastfeed their infant but find satisfaction in this rigorous process because of the maternal-infant bonding it promotes. The adoptive mother seeking to induce lactation is a unique client in need of highly tailored and personalized care.

  6. Lactation--how important is it?

    PubMed

    MacIntyre, U E; Walker, A R

    1994-02-01

    In Africa, with poverty rising, and health care diminishing, a nutritional question is-what are the limits of successful physiological adaptation to low dietary intakes? In this review the practice of lactation is discussed, from both past and present viewpoints. Orthodox Recommended Allowances for lactation are scarcely ever met. Yet, through various adaptations, the huge majority of African and similarly placed mothers lactate successfully and produce milk of good quantity and quality. The benefits from supplements are discussed; broadly, results have been disappointing. Some good and adverse non-dietary practices are considered. Benefits for infant and mother from lactation, even for protracted periods, far exceed drawbacks.

  7. Hemichannel-mediated release of lactate.

    PubMed

    Karagiannis, Anastassios; Sylantyev, Sergiy; Hadjihambi, Anna; Hosford, Patrick S; Kasparov, Sergey; Gourine, Alexander V

    2016-07-01

    In the central nervous system lactate contributes to the extracellular pool of readily available energy substrates and may also function as a signaling molecule which mediates communication between glial cells and neurons. Monocarboxylate transporters are believed to provide the main pathway for lactate transport across the membranes. Here we tested the hypothesis that lactate could also be released via opening of pannexin and/or functional connexin hemichannels. In acute slices prepared from the brainstem, hippocampus, hypothalamus and cortex of adult rats, enzymatic amperometric biosensors detected significant tonic lactate release inhibited by compounds, which block pannexin/connexin hemichannels and facilitated by lowering extracellular [Ca(2+)] or increased PCO2 Enhanced lactate release triggered by hypoxia was reduced by ∼50% by either connexin or monocarboxylate transporter blockers. Stimulation of Schaffer collateral fibers triggered lactate release in CA1 area of the hippocampus, which was facilitated in conditions of low extracellular [Ca(2+)], markedly reduced by blockade of connexin hemichannels and abolished by lactate dehydrogenase inhibitor oxamate. These results indicate that lactate transport across the membranes may occur via mechanisms other than monocarboxylate transporters. In the central nervous system, hemichannels may function as a conduit of lactate release, and this mechanism is recruited during hypoxia and periods of enhanced neuronal activity. © The Author(s) 2015.

  8. Hemichannel-mediated release of lactate

    PubMed Central

    Karagiannis, Anastassios; Sylantyev, Sergiy; Hadjihambi, Anna; Hosford, Patrick S; Kasparov, Sergey

    2015-01-01

    In the central nervous system lactate contributes to the extracellular pool of readily available energy substrates and may also function as a signaling molecule which mediates communication between glial cells and neurons. Monocarboxylate transporters are believed to provide the main pathway for lactate transport across the membranes. Here we tested the hypothesis that lactate could also be released via opening of pannexin and/or functional connexin hemichannels. In acute slices prepared from the brainstem, hippocampus, hypothalamus and cortex of adult rats, enzymatic amperometric biosensors detected significant tonic lactate release inhibited by compounds, which block pannexin/connexin hemichannels and facilitated by lowering extracellular [Ca2+] or increased PCO2. Enhanced lactate release triggered by hypoxia was reduced by ∼50% by either connexin or monocarboxylate transporter blockers. Stimulation of Schaffer collateral fibers triggered lactate release in CA1 area of the hippocampus, which was facilitated in conditions of low extracellular [Ca2+], markedly reduced by blockade of connexin hemichannels and abolished by lactate dehydrogenase inhibitor oxamate. These results indicate that lactate transport across the membranes may occur via mechanisms other than monocarboxylate transporters. In the central nervous system, hemichannels may function as a conduit of lactate release, and this mechanism is recruited during hypoxia and periods of enhanced neuronal activity. PMID:26661210

  9. The effects of changes in nutritional demand on gastrointestinal parasitism in lactating rats.

    PubMed

    Normanton, Heidi; Houdijk, Jos G M; Jessop, Neil S; Knox, Dave P; Kyriazakis, Ilias

    2007-01-01

    Lactating rats experience a breakdown of immunity to parasites, i.e. they carry larger worm burdens after re-infection compared to their non-lactating counterparts. Feeding high-protein foods to lactating rats results in reduced worm burdens. This could be attributed to changes in gastrointestinal environment or to overcoming effects of nutrient scarcity on host immunity. The latter hypothesis was addressed through a manipulation of nutrient demand by manipulating litter size. Twenty-three rats were immunized prior to mating and re-infected on day 2 of lactation with 1600 infective Nippostrongylus brasiliensis larvae. From parturition onwards, rats received ad libitum low-protein food (100 g crude protein/kg). Litter size were standardised to nine (LS9), six (LS6) or three (LS3) pups, by day 2 of lactation. After a further 10 d, LS9 and LS6 rats carried more worms than LS3 rats. However, feeding treatment did not affect concentrations of mucosal inflammatory cells. Achieved feed intake did not differ consistently between the treatment groups. However, LS9 and LS6 rats lost weight, whilst LS3 rats gained weight during lactation. The results support the view that resistance to N. brasiliensis is sensitive to changes in nutrient demand, and the improved resistance to N. brasiliensis is likely due to effects of overcoming nutrient scarcity on host immunity.

  10. Increased maternofetal calcium flux in parathyroid hormone-related protein-null mice

    PubMed Central

    Bond, H; Dilworth, M R; Baker, B; Cowley, E; Requena Jimenez, A; Boyd, R D H; Husain, S M; Ward, B S; Sibley, C P; Glazier, J D

    2008-01-01

    The role of parathyroid hormone-related protein (PTHrP) in fetal calcium homeostasis and placental calcium transport was examined in mice homozygous for the deletion of the PTHrP gene (PTHrP−/− null; NL) compared to PTHrP+/+ (wild-type; WT) and PTHrP+/− (heterozygous; HZ) littermates. Fetal blood ionized calcium was significantly reduced in NL fetuses compared to WT and HZ groups at 18 days of pregnancy (dp) with abolition of the fetomaternal calcium gradient. In situ placental perfusion of the umbilical circulation at 18 dp was used to measure unidirectional clearance of 45Ca across the placenta in maternofetal (CaKmf) and fetoplacental (CaKfp) directions; CaKfp was < 5% of CaKmf for all genotypes. At 18 dp, CaKmf across perfused placenta and intact placenta (CaKmf(intact)) were similar and concordant with net calcium accretion rates in vivo. CaKmf was significantly raised in NL fetuses compared to WT and HZ littermates. Calcium accretion was significantly elevated in NL fetuses by 19 dp. Placental calbindin-D9K expression in NL fetuses was marginally enhanced (P < 0.07) but expression of TRPV6/ECaC2 and plasma membrane Ca2+-ATPase (PMCA) isoforms 1 and 4 were unaltered. We conclude that PTHrP is an important regulator of fetal calcium homeostasis with its predominant effect being on unidirectional maternofetal transfer, probably mediated by modifying placental calbindin-D9K expression. In situ perfusion of mouse placenta is a robust methodology for allowing detailed dissection of placental transfer mechanisms in genetically modified mice. PMID:18258656

  11. Lactate Sensors on Flexible Substrates

    PubMed Central

    Yang, Xuesong; Fu, Timothy; Kota, Pavan Kumar; Tjia, Maggie; Nguyen, Cuong Manh; Chiao, Jung-Chih

    2016-01-01

    Lactate detection by an in situ sensor is of great need in clinical medicine, food processing, and athletic performance monitoring. In this paper, a flexible, easy to fabricate, and low-cost biosensor base on lactate oxidase is presented. The fabrication processes, including metal deposition, sol-gel IrOx deposition, and drop-dry enzyme loading method, are described in detail. The loaded enzyme was examined by scanning electron microscopy. Cyclic voltammetry was used to characterize the sensors. Durability, sensibility, and selectivity of the biosensors were examined. The comparison for different electrode sizes and different sensing film materials was conducted. The sensor could last for four weeks with an average surface area normalized sensitivity of 950 nA/(cm2 mM) and 9250 nA/(cm2 mM) for Au-based electrodes, and IrOx-modified electrodes respectively, both with an electrode size of 100 × 50 μm. The self-referencing method to record noises simultaneously with the working electrode greatly improved sensor sensitivity and selectivity. The sensor showed little response to interference chemicals, such as glutamate and dopamine. PMID:27657147

  12. Breastfeeding practices and lactation mastitis.

    PubMed

    Foxman, B; Schwartz, K; Looman, S J

    1994-03-01

    Clinical impression suggests that lactation mastitis is associated with inexperienced nursers, improper nursing techniques, stress and fatigue. A pilot study was conducted to describe the frequency of self-reported breastfeeding practices during the first week post partum among 100 breastfeeding women delivering at a freestanding birthing center or participating in an early discharge program. Nine cases of lactation mastitis were identified from the survey population and an additional 8 from the target population for the survey. Seventeen controls matched by delivery date were identified from survey participants. The frequency of self-reported breastfeeding practices, the presence of fatigue and stress during the week prior to the mastitis date in the case was compared among cases and controls. In the first week post partum, most women fed their babies every 2-3 hr for approx. 20 min a feeding. The cradle or Madonna position was the most frequently used nursing position. Nine percent reported supplementing feedings with formula. Women with mastitis were more likely than controls to report a history of mastitis with a previous child. In the week prior to the mastitis date of the case, women with mastitis were more likely than controls to report breast or nipple pain and cracks or breast fissures. They were less likely to report being able to take a daytime nap. Future studies should focus on the relative importance of and interrelationships among these factors.

  13. Leptin, GH, PRL, insulin and metabolic parameters throughout the dry period and lactation in dairy cows.

    PubMed

    Accorsi, P A; Govoni, N; Gaiani, R; Pezzi, C; Seren, E; Tamanini, C

    2005-06-01

    Leptin may play a role in the endocrine-metabolic processes that guarantee the physiological course of lactation in dairy cattle. This study was aimed at determining the changes in plasma concentrations of leptin and some of the main hormones and metabolites involved in the lactogenetic process in high-yielding dairy cows throughout lactation; we also wanted to assess whether leptin secretion is subjected to seasonal influences. Blood samples were collected from 23 Italian Friesian dairy cows from the end of a lactation to the ninth month of the subsequent one; in addition, blood was sampled from 47 dairy cows in different phases of lactation during February and July. Plasma concentrations of leptin, growth hormone (GH), insulin, prolactin (PRL), glucose, non-esterified fatty acids (NEFA) and urea were quantified by either validated radioimmunoassay (RIA) or enzymatic colorimetric methods. At the beginning of lactation, GH concentrations significantly increased, while a significant reduction occurred in leptin and insulin. This endocrine condition, such as the significant increase in NEFA plasma concentrations, is indicative of a marked lipid mobilization. In the more advanced stages of lactation, when both energy and protein balances become positive, leptin plasma concentrations increased, whereas GH and NEFA concentrations declined. During the summer months, a significant increase in leptin plasma concentrations, irrespective of the phase of lactation, was observed. Collectively, our findings suggest that, in dairy cows, leptin may represent a 'metabolic signal' of animal's status of fattening and nutritional level; in addition, leptin seems to be influenced by photoperiod and environmental temperature.

  14. Composition of milk obtained from unmassaged versus massaged breasts of lactating mothers.

    PubMed

    Foda, Mervat I; Kawashima, Takaaki; Nakamura, Sadako; Kobayashi, Michiko; Oku, Tsuneyuki

    2004-05-01

    The Oketani method is a program of breast massage and clinical counseling developed by the midwife Satomi Oketani. The purpose of this study is to examine the effects of the method on the quality of breast milk by determining the chemical composition of the milk before and after massage. Milk samples were obtained immediately before and after massage from healthy, exclusively breast-feeding Japanese mothers at two different periods of lactation one <3 months the other >3 months after parturition. Lipids, whey protein, casein, lactose, ash, and total solids in milk were measured in milk samples. The gross energy content of milk was estimated. Breast massage significantly increased lipids in the late lactating period but not in the early lactating period. In the early lactating period casein was increased by breast massage but was not significantly affected in the late lactating period. Breast massage caused a significant increase in total solids from the first day to 11 months post partum. The gross energy in the late lactating period was significantly increased by breast massage but not in the early lactating period. Lactose was not significantly changed by breast massage. Breast massage improves the quality of human milk by significantly increasing total solids, lipids, and casein concentration and gross energy. The milk of mothers treated by Oketani breast massage may improve the growth and development of infants.

  15. Reference values of blood parameters in beef cattle of different ages and stages of lactation.

    PubMed Central

    Doornenbal, H; Tong, A K; Murray, N L

    1988-01-01

    Reference (normal) values for 12 blood serum components were determined for 48 Shorthorn cows (2-10 years old) and their 48 calves, 357 crossbred cows (12-14 years old), 36 feedlot bulls and 36 feedlot steers. In addition, hemoglobin, hematocrit, triiodothyronine, thyroxine and cortisol levels were determined for the crossbred cows, and feedlot bulls and steers. Reference values were tabulated according to sex, age and stage of lactation. Serum concentrations of urea, total protein and bilirubin, and serum activity of aspartate aminotransferase and lactate dehydrogenase increased with age (P less than 0.05), while calcium, phosphorus and alkaline phosphatase decreased with age (P less than 0.05) from birth to the age of ten years. The Shorthorn cows had the highest levels of glucose at parturition (P less than 0.05) with decreasing levels during lactation. Creatinine concentration decreased during lactation and increased during postweaning. Both lactate dehydrogenase and aspartate aminotransferase levels increased (P less than 0.05) during lactation. Urea and uric acid were present at higher concentrations in lactating than nonlactating cows (P less than 0.05). The values reported, based on a wide age range and large number of cattle, could serve as clinical guides and a basis for further research. PMID:3349406

  16. Lactate and glucose interactions during rest and exercise in men: effect of exogenous lactate infusion.

    PubMed

    Miller, Benjamin F; Fattor, Jill A; Jacobs, Kevin A; Horning, Michael A; Navazio, Franco; Lindinger, Michael I; Brooks, George A

    2002-11-01

    To test the hypothesis that lactate plays a central role in the distribution of carbohydrate (CHO) potential energy for oxidation and glucose production (GP), we performed a lactate clamp (LC) procedure during rest and moderate intensity exercise. Blood [lactate] was clamped at approximately 4 mM by exogenous lactate infusion. Subjects performed 90 min exercise trials at 65 % of the peak rate of oxygen consumption (V(O(2))(,peak); 65 %), 55 % V(O(2))(,peak) (55 %) and 55 % V(O(2))(,peak) with lactate clamped to the blood [lactate] that was measured at 65 % V(O(2))(,peak) (55 %-LC). Lactate and glucose rates of appearance (R(a)), disappearance (R(d)) and oxidation (R(ox)) were measured with a combination of [3-(13)C]lactate, H(13)CO(3)(-), and [6,6-(2)H(2)]glucose tracers. During rest and exercise, lactate R(a) and R(d) were increased at 55 %-LC compared to 55 %. Glucose R(a) and R(d) were decreased during 55 %-LC compared to 55 %. Lactate R(ox) was increased by LC during exercise (55 %: 6.52 +/- 0.65 and 55 %-LC: 10.01 +/- 0.68 mg kg(-1) min(-1)) which was concurrent with a decrease in glucose oxidation (55 %: 7.64 +/- 0.4 and 55 %-LC: 4.35 +/- 0.31 mg kg(-1) min(-1)). With LC, incorporation of (13)C from tracer lactate into blood glucose (L GNG) increased while both GP and calculated hepatic glycogenolysis (GLY) decreased. Therefore, increased blood [lactate] during moderate intensity exercise increased lactate oxidation, spared blood glucose and decreased glucose production. Further, exogenous lactate infusion did not affect rating of perceived exertion (RPE) during exercise. These results demonstrate that lactate is a useful carbohydrate in times of increased energy demand.

  17. Yeast cell-based analysis of human lactate dehydrogenase isoforms.

    PubMed

    Mohamed, Lulu Ahmed; Tachikawa, Hiroyuki; Gao, Xiao-Dong; Nakanishi, Hideki

    2015-12-01

    Human lactate dehydrogenase (LDH) has attracted attention as a potential target for cancer therapy and contraception. In this study, we reconstituted human lactic acid fermentation in Saccharomyces cerevisiae, with the goal of constructing a yeast cell-based LDH assay system. pdc null mutant yeast (mutated in the endogenous pyruvate decarboxylase genes) are unable to perform alcoholic fermentation; when grown in the presence of an electron transport chain inhibitor, pdc null strains exhibit a growth defect. We found that introduction of the human gene encoding LDHA complemented the pdc growth defect; this complementation depended on LDHA catalytic activity. Similarly, introduction of the human LDHC complemented the pdc growth defect, even though LDHC did not generate lactate at the levels seen with LDHA. In contrast, the human LDHB did not complement the yeast pdc null mutant, although LDHB did generate lactate in yeast cells. Expression of LDHB as a red fluorescent protein (RFP) fusion yielded blebs in yeast, whereas LDHA-RFP and LDHC-RFP fusion proteins exhibited cytosolic distribution. Thus, LDHB exhibits several unique features when expressed in yeast cells. Because yeast cells are amenable to genetic analysis and cell-based high-throughput screening, our pdc/LDH strains are expected to be of use for versatile analyses of human LDH. © The Authors 2015. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

  18. Acute mercury exposition of virgin, pregnant, and lactating rats: Histopathological kidney and liver evaluations.

    PubMed

    Oliveira, Vitor Antunes; Favero, Gaia; Stacchiotti, Alessandra; Giugno, Lorena; Buffoli, Barbara; de Oliveira, Claudia Sirlene; Lavazza, Antonio; Albanese, Massimo; Rodella, Luigi Fabrizio; Pereira, Maria Ester; Rezzani, Rita

    2017-05-01

    This work investigated the effects of mercury chloride (HgCl2 ) acute exposure on virgin, pregnant and lactating rats by determination of renal and hepatic morphological and ultrastructural parameters and the expression of oxidative stress and stress tolerance markers, due to kidney and liver are the organs that more accumulate inorganic mercury. Adult Wistar rats virgin (90 days old), pregnant (18(th) gestation day) and lactating (7(th) lactation day) were injected once with HgCl2 (5 mg/kg) or saline (controls). We observed that HgCl2 exposure of virgin rats caused significant inflammatory infiltration and severe morphological variations, like glomeruli atrophy, dilatation of Bowman's capsule, tubular degeneration and hepatocytes alteration. Moreover, virgin rats presented mitochondrial modification, important oxidative stress and increase in stress tolerance proteins at both kidney and liver level, compared with virgin controls. In detail, virgin rats exposed to HgCl2 presented significantly elevated level of inducible nitric oxide synthase, heat shock protein 27 and glucose regulated proteins 75 expressions at both renal tubular and hepatocytes level, respect untreated virgin rats. Interestingly, pregnant and lactating rats exposed to HgCl2 presented weak renal and liver morphological alterations, showing weak inflammatory infiltration and no significant difference in structural mitochondrial transmembrane protein, oxidative stress markers and stress tolerance proteins expressions respect controls (virgin, pregnant and lactating rats). Although, both control and HgCl2 -exposed pregnant and lactating rats showed renal glomeruli greater in diameter respect virgin rats. In conclusion, we believe that virgin rats are more sensitive to HgCl2 toxicity respect pregnant and lactating rats. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 1500-1512, 2017. © 2016 Wiley Periodicals, Inc.

  19. Mechanism of lactate-induced relaxation of isolated rat mesenteric resistance arteries.

    PubMed Central

    McKinnon, W; Aaronson, P I; Knock, G; Graves, J; Poston, L

    1996-01-01

    1. The effects of the sodium salt of the weak acid lactate on tension and intracellular pH (pH1) were studied in rat mesenteric small arteries mounted on a wire myograph. Sodium lactate was substituted iso-osmotically for sodium chloride. 2. At a concentration of 50 mM, both L- and D-stereoisomers of lactate markedly relaxed arteries preconstricted with noradrenaline (NA) within 10 min. The concentration-response relationship for L-lactate showed that the NA contracture was relaxed by 50% at approximately 26 mM. L-Lactate did not, however, relax arteries preconstricted with high-K+(45 mM) solution. 3. L-Lactate did not alter extracellular pH (pHo) but caused a small but significant decrease in pH1, measured using the pH-sensitive fluorochrome, 2',7'-bis(carboxyethyl)-5-(6)-carboxyfluorescein (BCECF). Relaxation to L-lactate was unaffected when this change in pHi was offset by the simultaneous addition of NH4Cl to the solution. 4. Sodium pyruvate (50 mM) caused a significant intracellular acidosis but did not relax arteries preconstricted with NA. 5. L-Lactate-induced relaxations were unaffected by removal of the endothelium or when the synthesis of nitric oxide (NO) was inhibited by 10(-4) M N omega-nitro-L-arginine methyl ester (L-NAME). 6. The potassium channel blockers glibenclamide (10 microM), 4-aminopyridine (3 mM) and tetraethylammonium chloride (10 mM) did not affect L-lactate-induced relaxation in arteries preconstricted with NA. Inhibition of guanylate cyclase with Methylene Blue, or cyclooxgenase with indomethacin, also did not affect relaxation to L-lactate. 7. The Rp stereoisomer of adenosine-3',5'-cyclic monophosphothioate (Rp-cAMPS), an analogue of cAMP which inhibits competitively stimulation of protein kinase A, reduced significantly L-lactate-induced relaxation at a concentration of 25 microM. Rp-cAMPS also significantly reduced forskolin-induced relaxation of the NA contracture. 8. It is concluded that L-lactate-induced relaxation in this

  20. Lactate oxidation in human skeletal muscle mitochondria.

    PubMed

    Jacobs, Robert A; Meinild, Anne-Kristine; Nordsborg, Nikolai B; Lundby, Carsten

    2013-04-01

    Lactate is an important intermediate metabolite in human bioenergetics and is oxidized in many different tissues including the heart, brain, kidney, adipose tissue, liver, and skeletal muscle. The mechanism(s) explaining the metabolism of lactate in these tissues, however, remains unclear. Here, we analyze the ability of skeletal muscle to respire lactate by using an in situ mitochondrial preparation that leaves the native tubular reticulum and subcellular interactions of the organelle unaltered. Skeletal muscle biopsies were obtained from vastus lateralis muscle in 16 human subjects. Samples were chemically permeabilized with saponin, which selectively perforates the sarcolemma and facilitates the loss of cytosolic content without altering mitochondrial membranes, structure, and subcellular interactions. High-resolution respirometry was performed on permeabilized muscle biopsy preparations. By use of four separate and specific substrate titration protocols, the respirometric analysis revealed that mitochondria were capable of oxidizing lactate in the absence of exogenous LDH. The titration of lactate and NAD(+) into the respiration medium stimulated respiration (P ≤ 0.003). The addition of exogenous LDH failed to increase lactate-stimulated respiration (P = 1.0). The results further demonstrate that human skeletal muscle mitochondria cannot directly oxidize lactate within the mitochondrial matrix. Alternately, these data support previous claims that lactate is converted to pyruvate within the mitochondrial intermembrane space with the pyruvate subsequently taken into the mitochondrial matrix where it enters the TCA cycle and is ultimately oxidized.

  1. Carrier-mediated L-lactate transport in brush-border membrane vesicles from rat placenta during late gestation.

    PubMed Central

    Alonso de la Torre, S R; Serrano, M A; Alvarado, F; Medina, J M

    1991-01-01

    The mechanism for L-lactate transport across microvillous membrane vesicles prepared from rat placenta was examined. Uptake of L-lactate into these vesicles was mainly the result of transport into the intravesicular (osmotically active) space. The initial rate of L-lactate uptake was not affected by the presence of an inward gradient of either Na+ or K+. In the presence of an inward-directed proton gradient, L-lactate uptake was markedly stimulated, accumulating at concentrations 6-7-fold higher than the equilibrium. Lower transmembrane pH gradients were associated with slower initial uptakes and smaller overshoots. L-Lactate uptake determined under an inside-directed pH gradient was strongly inhibited by p-chloromercuriphenylsulphonic acid, a protein-thiol oxidizing agent. L-Lactate uptake was: (1) saturable as a function of the concentration of L-lactate, (2) inhibited by monocarboxylic acids such as pyruvate, D-lactate, beta-hydroxybutyrate and alpha-cyano-4-hydroxycinnamic acid, and (3) temperature-dependent. When present inside the vesicles, L-lactate, pyruvate and beta-hydroxybutyrate caused trans-stimulation of L-lactate uptake both in the presence and in the absence of an inside-directed pH gradient, indicating that L-lactate transport is a reversible process that can be shared by other monocarboxylic acids. There were no significant changes in maximal initial rate or in the kinetic parameters of L-lactate transport during the last 3 days of gestation. PMID:1654886

  2. Possible mechanisms for lactate's induction of panic.

    PubMed

    Liebowitz, M R; Gorman, J M; Fyer, A; Dillon, D; Levitt, M; Klein, D F

    1986-04-01

    Forty-three patients with panic disorder or agoraphobia with panic attacks and 20 control subjects received 0.5 M racemic sodium lactate intravenous infusions, single-blind as to duration and sequence. During the procedure, pulse; blood pressure; blood L-lactate and pyruvate; plasma ionized calcium, phosphate, prolactin, epinephrine, norepinephrine, and cortisol levels; and venous PCO2, pH, and bicarbonate were measured in an attempt to clarify the mechanism of lactate-induced panic attacks. During the infusion, 72% of the patients but none of the control subjects had panic attacks. The laboratory findings suggest that peripheral catecholamine surge is not the mechanism by which lactate causes panic, although elevated epinephrine may be a predisposing factor. Heightened central noradrenergic activity was present in many but not all of the attacks. Contrary to previous hypotheses, neither depression of ionized calcium nor induction of metabolic alkalosis appears sufficient to cause panic during lactate infusion.

  3. Isolation and Expression of Lactate Dehydrogenase Genes from Rhizopus oryzae

    PubMed Central

    Skory, Christopher D.

    2000-01-01

    Rhizopus oryzae is used for industrial production of lactic acid, yet little is known about the genetics of this fungus. In this study I cloned two genes, ldhA and ldhB, which code for NAD+-dependent l-lactate dehydrogenases (LDH) (EC 1.1.1.27), from a lactic acid-producing strain of R. oryzae. These genes are similar to each other and exhibit more than 90% nucleotide sequence identity and they contain no introns. This is the first description of ldh genes in a fungus, and sequence comparisons revealed that these genes are distinct from previously isolated prokaryotic and eukaryotic ldh genes. Protein sequencing of the LDH isolated from R. oryzae during lactic acid production confirmed that ldhA codes for a 36-kDa protein that converts pyruvate to lactate. Production of LdhA was greatest when glucose was the carbon source, followed by xylose and trehalose; all of these sugars could be fermented to lactic acid. Transcripts from ldhB were not detected when R. oryzae was grown on any of these sugars but were present when R. oryzae was grown on glycerol, ethanol, and lactate. I hypothesize that ldhB encodes a second NAD+-dependent LDH that is capable of converting l-lactate to pyruvate and is produced by cultures grown on these nonfermentable substrates. Both ldhA and ldhB restored fermentative growth to Escherichia coli (ldhA pfl) mutants so that they grew anaerobically and produced lactic acid. PMID:10831409

  4. Treatment of mastitis during lactation

    PubMed Central

    2009-01-01

    Treatment of mastitis should be based on bacteriological diagnosis and take national and international guidelines on prudent use of antimicrobials into account. In acute mastitis, where bacteriological diagnosis is not available, treatment should be initiated based on herd data and personal experience. Rapid bacteriological diagnosis would facilitate the proper selection of the antimicrobial. Treating subclinical mastitis with antimicrobials during lactation is seldom economical, because of high treatment costs and generally poor efficacy. All mastitis treatment should be evidence-based, i.e., the efficacy of each product and treatment length should be demonstrated by scientific studies. Use of on-farm written protocols for mastitis treatment promotes a judicious use of antimicrobials and reduces the use of antimicrobials. PMID:22081939

  5. Structural Basis of the Lactate-dependent Allosteric Regulation of Oxygen Binding in Arthropod Hemocyanin*

    PubMed Central

    Hirota, Shun; Tanaka, Naoki; Mičetić, Ivan; Di Muro, Paolo; Nagao, Satoshi; Kitagishi, Hiroaki; Kano, Koji; Magliozzo, Richard S.; Peisach, Jack; Beltramini, Mariano; Bubacco, Luigi

    2010-01-01

    Hemocyanin (Hc) is an oxygen carrier protein in which oxygen binding is regulated by allosteric effectors such as H+ and l-lactate. Isothermal titration calorimetric measurements showed that l-lactate binds to dodecameric and heterohexameric Hc and to the CaeSS3 homohexamer but not to the CaeSS2 monomer. The binding of lactate caused no change in the optical absorption and x-ray absorption spectra of either oxy- or deoxy-Hc, suggesting that no structural rearrangement of the active site occurred. At pH 6.5, the oxygen binding rate constant kobs obtained by flash photolysis showed a significant increase upon addition of l-lactate, whereas l-lactate addition had little effect at pH 8.3. Lactate binding caused a concentration-dependent shift in the interhexameric distances at pH 6.5 based on small angle x-ray scattering measurements. These results show that l-lactate affects oxygen affinity at pH 6.5 by modulating the global structure of Hc without affecting its binuclear copper center (the active site). In contrast to this, the active site structure of deoxy-Hc is affected by changes in pH (Hirota, S., Kawahara, T., Beltramini, M., Di Muro, P., Magliozzo, R. S., Peisach, J., Powers, L. S., Tanaka, N., Nagao, S., and Bubacco, L. (2008) J. Biol. Chem. 283, 31941–31948). Upon addiction of lactate, the kinetic behavior of oxygen rebinding for Hc was heterogeneous under low oxygen concentrations at pH 6.5 due to changes in the T and R state populations, and the equilibrium was found to shift from the T toward the R state with addition of lactate. PMID:20406810