Equilibrium and nonequilibrium fluctuations at the interface between two fluid phases

We have performed small-angle light-scattering measurements of the static structure factor of a critical binary mixture undergoing diffusive partial remixing. An uncommon scattering geometry integrates the structure factor over the sample thickness, allowing different regions of the concentration profile to be probed simultaneously. Our experiment shows the existence of interface capillary waves throughout the macroscopic evolution to an equilibrium interface, and allows to derive the time evolution of surface tension. Interfacial properties are shown to attain their equilibrium values quickly compared to the system's macroscopic equilibration time.Comment: 10 pages, 5 figures, submitted to PR

Shear-induced quench of long-range correlations in a liquid mixture

A static correlation function of concentration fluctuations in a (dilute) binary liquid mixture subjected to both a concentration gradient and uniform shear flow is investigated within the framework of fluctuating hydrodynamics. It is shown that a well-known $|\nabla c|^2/k^4$ long-range correlation at large wave numbers $k$ crosses over to a weaker divergent one for wave numbers satisfying $k<(\dot{\gamma}/D)^{1/2}$, while an asymptotic shear-controlled power-law dependence is confirmed at much smaller wave numbers given by $k\ll (\dot{\gamma}/\nu)^{1/2}$, where $c$, $\dot{\gamma}$, $D$ and $\nu$ are the mass concentration, the rate of the shear, the mass diffusivity and the kinematic viscosity of the mixture, respectively. The result will provide for the first time the possibility to observe the shear-induced suppression of a long-range correlation experimentally by using, for example, a low-angle light scattering technique.Comment: 8pages, 2figure

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

[EN] Mechanisms controlling immune function of dairy cows are dysregulated during heat stress (HS). Methyl donor supply-methionine (Met) and choline (Chop-positively modulates innate immune function, particularly antioxidant systems of polymorphonuclear leukocytes (PMN). The objective of this study was to investigate the effect of Met and Chol supply in vitro on mRNA abundance of genes related to 1-carbon metabolism, inflammation, and immune function in short-term cultures of PMN isolated from mid-lactating Holstein cows in response to heat challenge. Blood PMN were isolated from 5 Holstein cows (153 +/- 5 d postpartum, 34.63 +/- 2.73 kg/d of milk production; mean +/- SD). The PMN were incubated for 2 h at thermal-neutral (37 degrees C; TN) or heat stress (42 degrees C; HS) temperatures with 3 levels of Chol (0, 400, or 800 mu g/mL) or 3 ratios of Lys:Met (Met; 3.6:1, 2.9:1, or 2.4:1). Supernatant concentrations of IL-1 beta, IL-6, and tumor necrosis factor-alpha were measured via bovine-specific ELISA. Fold-changes in mRNA abundance were calculated separately for Chol and Met treatments to obtain the fold-change response at 42 degrees C (HS) relative to 37 degrees C (TN). Data were subjected to ANOVA using PROC MIXED in SAS (SAS Institute Inc., Cary, NC). Orthogonal contrasts were used to determine the linear or quadratic effect of Met and Chol for mRNA fold-change and supernatant cytokine concentrations. Compared with PMN receiving 0 mu g of Chol/mL, heat-stressed PMN supplemented with Chol at 400 or 800 mu g/mL had greater fold-change in abundance of CBS, CSAD, GSS, GSR, and GPX1. Among genes associated with inflammation and immune function, fold-change in abundance of TLR2, TLR4, IRAK1, IL1B, and IL10 increased with 400 and 800 mu g of Chol/mL compared with PMN receiving 0 mu g of Chol/mL. Fold-change in abundance of SAHH decreased linearly at increasing levels of Met supply. A linear effect was detected for MPO, NFKB1, and SOD1 due to greater fold-change in abundance when Met was increased to reach Lys: Met ratios of 2.9:1 and 2.4:1. Although increasing Chol supply upregulated BAX, BCL2, and HSP70, increased Met supply only upregulated BAX. Under HS conditions, enhancing PMN supply of Chol to 400 mu g/mL effectively increased fold-change in abundance of genes involved in antioxidant production (conferring cellular processes protection from free radicals and reactive oxygen species), inflammatory signaling, and innate immunity. Although similar outcomes were obtained with Met supply at Lys:Met ratios of 2.9:1 and 2.4:1, the response was less pronounced. Both Chol and Met supply enhanced the cytoprotective characteristics of PMN through upregulation of heat shock proteins. Overall, the modulatory effects detected in the present experiment highlight an opportunity to use Met and particularly Chol supplementation during thermal stress.M. Vailati-Riboni was supported in part by Hatch funds under project ILLU-538-914, National Institute of Food and Agriculture (Washington, DC). The authors declare no conflict of interest.Lopreiato, V.; Vailati-Riboni, M.; Parys, C.; Fernández Martínez, CJ.; Minuti, A.; Loor, J. (2020). Methyl donor supply to heat stress-challenged polymorphonuclear leukocytes from lactating Holstein cows enhances 1-carbon metabolism, immune response, and cytoprotective gene network abundance. Journal of Dairy Science. 103(11):10477-10493. https://doi.org/10.3168/jds.2020-18638S104771049310311Abdelmegeid, M. K., Vailati-Riboni, M., Alharthi, A., Batistel, F., & Loor, J. J. (2017). Supplemental methionine, choline, or taurine alter in vitro gene network expression of polymorphonuclear leukocytes from neonatal Holstein calves. Journal of Dairy Science, 100(4), 3155-3165. doi:10.3168/jds.2016-12025Armentano, L. E., Bertics, S. J., & Ducharme, G. A. (1997). Response of Lactating Cows to Methionine or Methionine Plus Lysine Added to High Protein Diets Based on Alfalfa and Heated Soybeans. Journal of Dairy Science, 80(6), 1194-1199. doi:10.3168/jds.s0022-0302(97)76047-8Banerjee, R., Evande, R., Kabil, Ö., Ojha, S., & Taoka, S. (2003). Reaction mechanism and regulation of cystathionine β-synthase. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, 1647(1-2), 30-35. doi:10.1016/s1570-9639(03)00044-xBatistel, F., Arroyo, J. M., Bellingeri, A., Wang, L., Saremi, B., Parys, C., … Loor, J. J. (2017). Ethyl-cellulose rumen-protected methionine enhances performance during the periparturient period and early lactation in Holstein dairy cows. Journal of Dairy Science, 100(9), 7455-7467. doi:10.3168/jds.2017-12689Baumgard, L. H., & Rhoads, R. P. (2013). Effects of Heat Stress on Postabsorptive Metabolism and Energetics. Annual Review of Animal Biosciences, 1(1), 311-337. doi:10.1146/annurev-animal-031412-103644Bernabucci, U., Biffani, S., Buggiotti, L., Vitali, A., Lacetera, N., & Nardone, A. (2014). The effects of heat stress in Italian Holstein dairy cattle. Journal of Dairy Science, 97(1), 471-486. doi:10.3168/jds.2013-6611Bernabucci, U., Lacetera, N., Baumgard, L. H., Rhoads, R. P., Ronchi, B., & Nardone, A. (2010). Metabolic and hormonal acclimation to heat stress in domesticated ruminants. Animal, 4(7), 1167-1183. doi:10.1017/s175173111000090xBoldyrev, A., Bryushkova, E., Mashkina, A., & Vladychenskaya, E. (2013). Why Is Homocysteine Toxic for the Nervous and Immune Systems? Current Aging Science, 6(1), 29-36. doi:10.2174/18746098112059990007Catozzi, C., Ávila, G., Zamarian, V., Pravettoni, D., Sala, G., Ceciliani, F., … Lecchi, C. (2020). In-vitro effect of heat stress on bovine monocytes lifespan and polarization. Immunobiology, 225(2), 151888. doi:10.1016/j.imbio.2019.11.023Chinenov, Y., Gupte, R., & Rogatsky, I. (2013). Nuclear receptors in inflammation control: Repression by GR and beyond. Molecular and Cellular Endocrinology, 380(1-2), 55-64. doi:10.1016/j.mce.2013.04.006Chorąży, M., Kontny, E., Marcinkiewicz, J., & Maśliński, W. (2002). Amino Acids, 23(4), 407-413. doi:10.1007/s00726-002-0204-0Coleman, D. N., Lopreiato, V., Alharthi, A., & Loor, J. J. (2020). Amino acids and the regulation of oxidative stress and immune function in dairy cattle. Journal of Animal Science, 98(Supplement_1), S175-S193. doi:10.1093/jas/skaa138Collier, R. J., Stiening, C. M., Pollard, B. C., VanBaale, M. J., Baumgard, L. H., Gentry, P. C., & Coussens, P. M. (2006). Use of gene expression microarrays for evaluating environmental stress tolerance at the cellular level in cattle1. Journal of Animal Science, 84(suppl_13), E1-E13. doi:10.2527/2006.8413_supple1xCouper, K. N., Blount, D. G., & Riley, E. M. (2008). IL-10: The Master Regulator of Immunity to Infection. The Journal of Immunology, 180(9), 5771-5777. doi:10.4049/jimmunol.180.9.5771Del Vesco, A. P., Gasparino, E., Grieser, D. de O., Zancanela, V., Soares, M. A. M., & de Oliveira Neto, A. R. (2015). Effects of methionine supplementation on the expression of oxidative stress-related genes in acute heat stress-exposed broilers. British Journal of Nutrition, 113(4), 549-559. doi:10.1017/s0007114514003535Ekremoğlu, M., Türközkan, N., Erdamar, H., Kurt, Y., & Yaman, H. (2006). Protective effect of taurine on respiratory burst activity of polymorphonuclear leukocytes in endotoxemia. Amino Acids, 32(3), 413-417. doi:10.1007/s00726-006-0382-2El-Benna, J., Hurtado-Nedelec, M., Marzaioli, V., Marie, J.-C., Gougerot-Pocidalo, M.-A., & Dang, P. M.-C. (2016). Priming of the neutrophil respiratory burst: role in host defense and inflammation. Immunological Reviews, 273(1), 180-193. doi:10.1111/imr.12447Esposito, G., Irons, P. C., Webb, E. C., & Chapwanya, A. (2014). Interactions between negative energy balance, metabolic diseases, uterine health and immune response in transition dairy cows. Animal Reproduction Science, 144(3-4), 60-71. doi:10.1016/j.anireprosci.2013.11.007Fear, J. M., & Hansen, P. J. (2011). Developmental Changes in Expression of Genes Involved in Regulation of Apoptosis in the Bovine Preimplantation Embryo1. Biology of Reproduction, 84(1), 43-51. doi:10.1095/biolreprod.110.086249Gao, S. T., Guo, J., Quan, S. Y., Nan, X. M., Fernandez, M. V. S., Baumgard, L. H., & Bu, D. P. (2017). The effects of heat stress on protein metabolism in lactating Holstein cows. Journal of Dairy Science, 100(6), 5040-5049. doi:10.3168/jds.2016-11913Han, Z.-Y., Mu, T., & Yang, Z. (2014). Methionine protects against hyperthermia-induced cell injury in cultured bovine mammary epithelial cells. Cell Stress and Chaperones, 20(1), 109-120. doi:10.1007/s12192-014-0530-7Heiser, A., LeBlanc, S. J., & McDougall, S. (2018). Pegbovigrastim treatment affects gene expression in neutrophils of pasture-fed, periparturient cows. Journal of Dairy Science, 101(9), 8194-8207. doi:10.3168/jds.2017-14129Horowitz, M. (2001). Heat acclimation: phenotypic plasticity and cues to the underlying molecular mechanisms. Journal of Thermal Biology, 26(4-5), 357-363. doi:10.1016/s0306-4565(01)00044-4Hunter-Lavin, C., Davies, E. L., Bacelar, M. M. F. V. G., Marshall, M. J., Andrew, S. M., & Williams, J. H. H. (2004). Hsp70 release from peripheral blood mononuclear cells. Biochemical and Biophysical Research Communications, 324(2), 511-517. doi:10.1016/j.bbrc.2004.09.075Ingvartsen, K. L., & Moyes, K. (2013). Nutrition, immune function and health of dairy cattle. Animal, 7, 112-122. doi:10.1017/s175173111200170xJoshi, B. C., Joshi, H. B., McDowell, R. E., & Sadhu, D. P. (1968). Composition of Skin Secretions from Three Indian Breeds of Cattle Under Thermal Stress. Journal of Dairy Science, 51(6), 917-920. doi:10.3168/jds.s0022-0302(68)87105-xKobayashi, S. D., & DeLeo, F. R. (2009). Role of neutrophils in innate immunity: a systems biology‐level approach. Wiley Interdisciplinary Reviews: Systems Biology and Medicine, 1(3), 309-333. doi:10.1002/wsbm.32Kumar, H., Kawai, T., & Akira, S. (2011). Pathogen Recognition by the Innate Immune System. International Reviews of Immunology, 30(1), 16-34. doi:10.3109/08830185.2010.529976Lacetera, N., Bernabucci, U., Basiricò, L., Morera, P., & Nardone, A. (2009). Heat shock impairs DNA synthesis and down-regulates gene expression for leptin and Ob-Rb receptor in concanavalin A-stimulated bovine peripheral blood mononuclear cells. Veterinary Immunology and Immunopathology, 127(1-2), 190-194. doi:10.1016/j.vetimm.2008.09.020Lacetera, N., Bernabucci, U., Scalia, D., Basiricò, L., Morera, P., & Nardone, A. (2006). Heat Stress Elicits Different Responses in Peripheral Blood Mononuclear Cells from Brown Swiss and Holstein Cows. Journal of Dairy Science, 89(12), 4606-4612. doi:10.3168/jds.s0022-0302(06)72510-3Lecchi, C., Rota, N., Vitali, A., Ceciliani, F., & Lacetera, N. (2016). In vitro assessment of the effects of temperature on phagocytosis, reactive oxygen species production and apoptosis in bovine polymorphonuclear cells. Veterinary Immunology and Immunopathology, 182, 89-94. doi:10.1016/j.vetimm.2016.10.007Loos, H., Roos, D., Weening, R., & Houwerzijl, J. (1976). Familial deficiency of glutathione reductase in human blood cells. Blood, 48(1), 53-62. doi:10.1182/blood.v48.1.53.53Lopreiato, V., Vailati-Riboni, M., Bellingeri, A., Khan, I., Farina, G., Parys, C., & Loor, J. J. (2019). Inflammation and oxidative stress transcription profiles due to in vitro supply of methionine with or without choline in unstimulated blood polymorphonuclear leukocytes from lactating Holstein cows. Journal of Dairy Science, 102(11), 10395-10410. doi:10.3168/jds.2019-16413Lubos, E., Loscalzo, J., & Handy, D. E. (2011). Glutathione Peroxidase-1 in Health and Disease: From Molecular Mechanisms to Therapeutic Opportunities. Antioxidants & Redox Signaling, 15(7), 1957-1997. doi:10.1089/ars.2010.3586Lushchak, V. I. (2012). Glutathione Homeostasis and Functions: Potential Targets for Medical Interventions. Journal of Amino Acids, 2012, 1-26. doi:10.1155/2012/736837McGuire, M. A., Beede, D. K., DeLorenzo, M. A., Wilcox, C. J., Huntington, G. B., Reynolds, C. K., & Collier, R. J. (1989). Effects of Thermal Stress and Level of Feed Intake on Portal Plasma Flow and Net Fluxes of Metabolites in Lactating Holstein Cows2,3. Journal of Animal Science, 67(4), 1050-1060. doi:10.2527/jas1989.6741050xMin, L., Zheng, N., Zhao, S., Cheng, J., Yang, Y., Zhang, Y., … Wang, J. (2016). Long-term heat stress induces the inflammatory response in dairy cows revealed by plasma proteome analysis. Biochemical and Biophysical Research Communications, 471(2), 296-302. doi:10.1016/j.bbrc.2016.01.185Moyes, K. M., Drackley, J. K., Morin, D. E., & Loor, J. J. (2010). Greater expression of TLR2, TLR4, and IL6 due to negative energy balance is associated with lower expression of HLA-DRA and HLA-A in bovine blood neutrophils after intramammary mastitis challenge with Streptococcus uberis. Functional & Integrative Genomics, 10(1), 53-61. doi:10.1007/s10142-009-0154-7Moyes, K. M., Graugnard, D. E., Khan, M. J., Mukesh, M., & Loor, J. J. (2014). Postpartal immunometabolic gene network expression and function in blood neutrophils are altered in response to prepartal energy intake and postpartal intramammary inflammatory challenge. Journal of Dairy Science, 97(4), 2165-2177. doi:10.3168/jds.2013-7433Nakamura, M. (2000). Preconditioning decreases Bax expression, PMN accumulation and apoptosis in reperfused rat heart. Cardiovascular Research, 45(3), 661-670. doi:10.1016/s0008-6363(99)00393-4Oeckinghaus, A., & Ghosh, S. (2009). The NF- B Family of Transcription Factors and Its Regulation. Cold Spring Harbor Perspectives in Biology, 1(4), a000034-a000034. doi:10.1101/cshperspect.a000034Osorio, J. S., Ji, P., Drackley, J. K., Luchini, D., & Loor, J. J. (2014). Smartamine M and MetaSmart supplementation during the peripartal period alter hepatic expression of gene networks in 1-carbon metabolism, inflammation, oxidative stress, and the growth hormone–insulin-like growth factor 1 axis pathways. Journal of Dairy Science, 97(12), 7451-7464. doi:10.3168/jds.2014-8680Salama, A. A. K., Duque, M., Wang, L., Shahzad, K., Olivera, M., & Loor, J. J. (2019). Enhanced supply of methionine or arginine alters mechanistic target of rapamycin signaling proteins, messenger RNA, and microRNA abundance in heat-stressed bovine mammary epithelial cells in vitro. Journal of Dairy Science, 102(3), 2469-2480. doi:10.3168/jds.2018-15219Schell, M. T., Spitzer, A. L., Johnson, J. A., Lee, D., & Harris, H. W. (2005). Heat Shock Inhibits NF-kB Activation in a Dose- and Time-Dependent Manner. Journal of Surgical Research, 129(1), 90-93. doi:10.1016/j.jss.2005.05.025Silanikove, N. (2000). Effects of heat stress on the welfare of extensively managed domestic ruminants. Livestock Production Science, 67(1-2), 1-18. doi:10.1016/s0301-6226(00)00162-7Stankiewicz, A. R., Lachapelle, G., Foo, C. P. Z., Radicioni, S. M., & Mosser, D. D. (2005). Hsp70 Inhibits Heat-induced Apoptosis Upstream of Mitochondria by Preventing Bax Translocation. Journal of Biological Chemistry, 280(46), 38729-38739. doi:10.1074/jbc.m509497200Steel, G. J., Fullerton, D. M., Tyson, J. R., & Stirling, C. J. (2004). Coordinated Activation of Hsp70 Chaperones. Science, 303(5654), 98-101. doi:10.1126/science.1092287Sun, D., Chen, D., Du, B., & Pan, J. (2005). Heat Shock Response Inhibits NF-κB Activation and Cytokine Production in Murine Kupffer Cells. Journal of Surgical Research, 129(1), 114-121. doi:10.1016/j.jss.2005.05.028Taraktsoglou, M., Szalabska, U., Magee, D. A., Browne, J. A., Sweeney, T., Gormley, E., & MacHugh, D. E. (2011). Transcriptional profiling of immune genes in bovine monocyte-derived macrophages exposed to bacterial antigens. Veterinary Immunology and Immunopathology, 140(1-2), 130-139. doi:10.1016/j.vetimm.2010.12.002Trevisi, E., Jahan, N., Bertoni, G., Ferrari, A., & Minuti, A. (2015). Pro-Inflammatory Cytokine Profile in Dairy Cows: Consequences for New Lactation. Italian Journal of Animal Science, 14(3), 3862. doi:10.4081/ijas.2015.3862Tsan, M.-F., & Gao, B. (2004). Cytokine function of heat shock proteins. American Journal of Physiology-Cell Physiology, 286(4), C739-C744. doi:10.1152/ajpcell.00364.2003Vailati-Riboni, M., Zhou, Z., Jacometo, C. B., Minuti, A., Trevisi, E., Luchini, D. N., & Loor, J. J. (2017). Supplementation with rumen-protected methionine or choline during the transition period influences whole-blood immune response in periparturient dairy cows. Journal of Dairy Science, 100(5), 3958-3968. doi:10.3168/jds.2016-11812Yan, J., Meng, X., Wancket, L. M., Lintner, K., Nelin, L. D., Chen, B., … Liu, Y. (2012). Glutathione Reductase Facilitates Host Defense by Sustaining Phagocytic Oxidative Burst and Promoting the Development of Neutrophil Extracellular Traps. The Journal of Immunology, 188(5), 2316-2327. doi:10.4049/jimmunol.1102683Zhou, Z., Bulgari, O., Vailati-Riboni, M., Trevisi, E., Ballou, M. A., Cardoso, F. C., … Loor, J. J. (2016). Rumen-protected methionine compared with rumen-protected choline improves immunometabolic status in dairy cows during the peripartal period. Journal of Dairy Science, 99(11), 8956-8969. doi:10.3168/jds.2016-10986Zhou, Z., Ferdous, F., Montagner, P., Luchini, D. N., Corrêa, M. N., & Loor, J. J. (2018). Methionine and choline supply during the peripartal period alter polymorphonuclear leukocyte immune response and immunometabolic gene expression in Holstein cows. Journal of Dairy Science, 101(11), 10374-10382. doi:10.3168/jds.2018-14972Zhou, Z., Vailati-Riboni, M., Trevisi, E., Drackley, J. K., Luchini, D. N., & Loor, J. J. (2016). Better postpartal performance in dairy cows supplemented with rumen-protected methionine compared with choline during the peripartal period. Journal of Dairy Science, 99(11), 8716-8732. doi:10.3168/jds.2015-1052

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

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

A new approach to a powered knee prosthesis: Layering powered assistance onto strictly passive prosthesis behavior

This article describes a novel approach to the control of a powered knee prosthesis where the control system provides passive behavior for most activities and then provides powered assistance only for those activities that require them. The control approach presented here is based on the categorization of knee joint function during activities into four behaviors: resistive stance behavior, active stance behavior, ballistic swing, and non-ballistic swing. The approach is further premised on the assumption that healthy non-perturbed swing-phase is characterized by a ballistic swing motion, and therefore, a replacement of that function should be similarly ballistic. The control system utilizes a six-state finite-state machine, where each state provides different constitutive behaviors (concomitant with the four aforementioned knee behaviors) which are appropriate for a range of activities. Transitions between states and torque control within states is controlled by user motion, such that the control system provides, to the extent possible, knee torque behavior as a reaction to user motion, including for powered behaviors. The control system is demonstrated on a novel device that provides a sufficiently low impedance to enable a strictly passive ballistic swing-phase, while also providing sufficiently high torque to offer powered stance-phase knee-extension during activities such as step-over stair ascent. Experiments employing the knee and control system on an individual with transfemoral amputation are presented that compare the functionality of the power-supplemented nominally passive system with that of a conventional passive microprocessor-controlled knee prosthesis

Endoscopic removal of a right main bronchus glomus tumor

Glomus tumours in the respiratory tract are very rare. The majority of the reported cases have been surgically treated. An approach with rigid bronchoscopy to endobronchial lesions suspected to be carcinoid or other well vascularized tumours, as glomus tumor is, should be considered because it can allow a safe diagnosis and eventually be therapeutic avoiding more invasive and surgical procedures

Endoscopic removal of a right main bronchus glomus tumor

Glomus tumours in the respiratory tract are very rare. The majority of the reported cases have been surgically treated. An approach with rigid bronchoscopy to endobronchial lesions suspected to be carcinoid or other well vascularized tumours, as glomus tumor is, should be considered because it can allow a safe diagnosis and eventually be therapeutic avoiding more invasive and surgical procedures

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

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

EasyPrimer: user-friendly tool for pan-PCR/HRM primers design. Development of an HRM protocol on wzi gene for fast Klebsiella pneumoniae typing

In this work we present EasyPrimer, a user-friendly online tool developed to assist pan-PCR and High Resolution Melting (HRM) primer design. The tool finds the most suitable regions for primer design in a gene alignment and returns a clear graphical representation of their positions on the consensus sequence. EasyPrimer is particularly useful in difficult contexts, e.g. on gene alignments of hundreds of sequences and/or on highly variable genes. HRM analysis is an emerging method for fast and cost saving bacterial typing and an HRM scheme of six primer pairs on five Multi-Locus Sequence Type (MLST) genes is already available for Klebsiella pneumoniae. We validated the tool designing a scheme of two HRM primer pairs on the hypervariable gene wzi of Klebsiella pneumoniae and compared the two schemes. The wzi scheme resulted to have a discriminatory power comparable to the HRM MLST scheme, using only one third of primer pairs. Then we successfully used the wzi HRM primer scheme to reconstruct a Klebsiella pneumoniae nosocomial outbreak in few hours. The use of hypervariable genes reduces the number of HRM primer pairs required for bacterial typing allowing to perform cost saving, large-scale surveillance programs