Annual Ryegrass Pasture for Dairy Cows Receiving Total Mixed Ration

The inclusion of herbage in the diet of medium yielding dairy cows offered a total mixed ration (TMR) may be beneficial. This study, which involved mid lactation dairy cows, examined the effect of partial replacement of a TMR with annual temperate pasture. Treatments were ad libitum TMR (TMR100), 75% ad libitum TMR + ryegrass (Lolium multiflorum ‘Maximus’) (TMR75), and 50% ad libitum TMR + ryegrass (TMR50). Twelve multiparous Holstein and F1 Jersey × Holstein cows were divided into six homogeneous groups, taking account of milk production (26.6 ± 4.5 kg/day), days in milk (128 ± 50) and body weight (546 ± 31 kg). Treatments were compared in a replicated 3×3 Latin square design, comprising three 21-day periods (measurements during final 5 days). Cows on TMR75 and TMR50 strip grazed between morning and afternoon milking (7 h/day), with a target pre- and post-grazing sward height of 24 and 12 cm, respectively. Herbage DM intake was estimated as the difference between pre- and post-grazing herbage mass. The TMR and herbage had a crude protein content of 150 and 303 g/kg DM, and a NDF content of 366 and 495 g/kg DM, respectively. Herbage DM intake increased from 4.8 kg/day in TMR75 to 6.7 kg/day on TMR50. Total DM intake decreased from 19.4 kg/day (TMR100), to 18.1 and 15.9 kg/day (TMR75 and TMR50, respectively). Milk production, milk fat and milk protein content were similar between treatments, averaging 25.6 kg/day, 44.6 g/kg and 33.7 g/kg, respectively. The net energy for lactation (NEL) supply was 113, 104 and 92% of NEL requirements, for cows receiving TMR100, TMR75 and TMR50, respectively. Ryegrass pastures were able to replace up to 50% of TMR offered to mid lactation dairy cows without any adverse effects on milk production and milk composition

The seismic signature of Upper‐Mantle Plumes: application to the Northern East African Rift

Several seismic and numerical studies proposed that below, some hotspots upper‐mantle plumelets rise from a thermal boundary layer below 660 km depth, fed by a deeper plume source. We recently found tomographic evidence of multiple upper‐mantle upwellings, spaced by several 100 km, rising through the transition zone below the northern East African Rift. To better test this interpretation, we run 3‐D numerical simulations of mantle convection for Newtonian and non‐Newtonian rheologies, for both thermal instabilities rising from a lower boundary layer, and the destabilization of a thermal anomaly placed at the base of the box (700–800 km depth). The thermal structures are converted to seismic velocities using a thermodynamic approach. Resolution tests are then conducted for the same P and S data distribution and inversion parameters as our traveltime tomography. The Rayleigh Taylor models predict simultaneous plumelets in different stages of evolution rising from a hot layer located below the transition zone, resulting in seismic structure that looks more complex than the simple vertical cylinders that are often anticipated. From the wide selection of models tested, we find that the destabilization of a 200 °C, 100 km thick thermal anomaly with a non‐Newtonian rheology, most closely matches the magnitude and the spatial and temporal distribution of the anomalies below the rift. Finally, we find that for reasonable upper‐mantle viscosities, the synthetic plume structures are similar in scale and shape to the actual low‐velocity anomalies, providing further support for the existence of upper‐mantle plumelets below the northern East African Rift

Smart solutions for low-income buildings rehabilitation: international researches and experiences

[EN] Smart City concept briefly refers to a sustainable city where innovative Smart strategies will be adopted to an efficient management of resources flows and social interoperability. Aligned with the most relevant European research experiences, the paper describes the relations between an on-going research from the PDTA Dept., centered on a knowledge platform tool for energy-efficient interactive buildings design, and the contribution of the ELIH-MED project on the use of industrialized solutions and smart devices during the rehabilitation process in Mediterranean low-income housings. A pilot project in the Spanish climate context reveals the important role of Smart Monitoring Devices to encourage energy savings and tackle recognized needs through a participative process where stakeholders and beneficiaries are actively involved.[IT] Il termine Smart City può essere sinteticamente associato al concetto di città sostenibile dove strategie innovative intelligenti favoriscono la gestione efficiente del flusso di risorse e l' interoperabilità sociale. Con riferimento alle più significative esperienze di ricerca europee, il contributo approfondisce i legami tra una ricerca, attualmente in corso presso il Dipartimento PDTA, rivolta allo sviluppo di una piattaforma per la progettazione di edifici interattivi ed energeticamente efficienti, e il progetto ELIH-MED incentrato sull'uso di soluzioni industrializzate e dispositivi Smart nell¿ambito di interventi di riqualificazione di edifici sociali nel bacino del Mediterraneo. Un progetto pilota situato nel contesto climatico spagnolo mostra l'impatto derivante dall¿uso di soluzioni Smart nel favorire il risparmio energetico e rispondere alle esigenze degli utenti attraverso un processo partecipativo che coinvolge attivamente stakeholder e beneficiari.The ELIH-Med project represents a significant pilot experience that has allowed to identify and demonstrate, through pilot-scale actions, the feasibil-ity of efficient and innovative solutions and innovative financial mechanisms, backed by ERDF (European Regional Development Fund) funds.Arbizzani. E.; Civiero, P.; Ortega Madrigal, L.; Serrano Lanzarote, AB. (2015). Soluzioni Smart per la riqualificazione degli edifici residenziali sociali: ricerche ed esperienze internazionali. Techne: journal of technology for architecture and environment. 10:222-231. https://doi.org/10.13128/Techne-17519S2222311

RAB8, RAB10 and RILPL1 contribute to both LRRK2 kinase-mediated centrosomal cohesion and ciliogenesis deficits

Mutations in the LRRK2 kinase are the most common cause of familial Parkinson's disease, and variants increase risk for the sporadic form of the disease. LRRK2 phosphorylates multiple RAB GTPases including RAB8A and RAB10. Phosphorylated RAB10 is recruited to centrosome-localized RILPL1, which may interfere with ciliogenesis in a disease-relevant context. Our previous studies indicate that the centrosomal accumulation of phosphorylated RAB8A causes centrosomal cohesion deficits in dividing cells, including in peripheral patient-derived cells. Here, we show that both RAB8 and RAB10 contribute to the centrosomal cohesion deficits. Pathogenic LRRK2 causes the centrosomal accumulation not only of phosho-RAB8 but also of phospho-RAB10, and the effects on centrosomal cohesion are dependent on RAB8, RAB10 and RILPL1. Conversely, the pathogenic LRRK2-mediated ciliogenesis defects correlate with the centrosomal accumulation of both phospho-RAB8 and phospho-RAB10. LRRK2-mediated centrosomal cohesion and ciliogenesis alterations are observed in patient-derived peripheral cells, as well as in primary astrocytes from mutant LRRK2 mice, and are reverted upon LRRK2 kinase inhibition. These data suggest that the LRRK2-mediated centrosomal cohesion and ciliogenesis defects are distinct cellular readouts of the same underlying phospho-RAB8/RAB10/RILPL1 nexus and highlight the possibility that either centrosomal cohesion and/or ciliogenesis alterations may serve as cellular biomarkers for LRRK2-related PD

GTP binding controls complex formation by the human ROCO protein MASL1.

The human ROCO proteins are a family of multi-domain proteins sharing a conserved ROC-COR supra-domain. The family has four members: leucine-rich repeat kinase 1 (LRRK1), leucine-rich repeat kinase 2 (LRRK2), death-associated protein kinase 1 (DAPK1) and malignant fibrous histiocytoma amplified sequences with leucine-rich tandem repeats 1 (MASL1). Previous studies of LRRK1/2 and DAPK1 have shown that the ROC (Ras of complex proteins) domain can bind and hydrolyse GTP, but the cellular consequences of this activity are still unclear. Here, the first biochemical characterization of MASL1 and the impact of GTP binding on MASL1 complex formation are reported. The results demonstrate that MASL1, similar to other ROCO proteins, can bind guanosine nucleotides via its ROC domain. Furthermore, MASL1 exists in two distinct cellular complexes associated with heat shock protein 60, and the formation of a low molecular weight pool of MASL1 is modulated by GTP binding. Finally, loss of GTP enhances MASL1 toxicity in cells. Taken together, these data point to a central role for the ROC/GTPase domain of MASL1 in the regulation of its cellular function

BEL \u3b2-TREFOIL. A NOVEL LECTIN WITH ANTITUMORAL PROPERTIES IN KING BOLETE (BOLETUS EDULIS) MUSHROOMS

A novel lectin completely different from the formerly described member of the saline soluble family of mushroom specific lectins (named BEL, Boletus edulis lectin, Bovi et al., 2011) was purified from the fruiting bodies of king bolete mushrooms (also called porcino, cep or penny bun). The lectin was structurally characterized: its amino acid sequence and three dimensional structure were determined. The protein is a homodimer and each protomer folds as \u3b2-trefoil domain and therefore we propose the name BEL \u3b2-trefoil to distinguish it from the other lectin that has been described in these mushrooms: BEL. The new lectin has potent anti-proliferative effects on human epithelial cancer cells which confers to it an interesting therapeutic potential as an antineoplastic agent. Several crystal forms of the apoprotein and of complexes with different carbohydrates were studied by X-ray diffraction. The interaction of the lectin with lactose, galactose, N-acetylgalactosamine, T-antigen disaccharide (Gal\u3b21-3GalNAc) and T-antigen (Ser-Gal\u3b21-3GalNAc) was examined in detail. All the three potential binding sites present in the \u3b2-trefoil fold are occupied in at least one crystal form. No important conformational changes are observed in the lectin when comparing its co-crystals with carbohydrates with those of the ligand free protein

PAK6 Phosphorylates 14-3-3 gamma to Regulate Steady State Phosphorylation of LRRK2

Mutations in Leucine-rich repeat kinase 2 (LRRK2) are associated with Parkinson’s disease (PD) and, as such, LRRK2 is considered a promising therapeutic target for age-related neurodegeneration. Although the cellular functions of LRRK2 in health and disease are incompletely understood, robust evidence indicates that PD-associated mutations alter LRRK2 kinase and GTPase activities with consequent deregulation of the downstream signaling pathways. We have previously demonstrated that one LRRK2 binding partner is P21 (RAC1) Activated Kinase 6 (PAK6). Here, we interrogate the PAK6 interactome and find that PAK6 binds a subset of 14-3-3 proteins in a kinase dependent manner. Furthermore, PAK6 efficiently phosphorylates 14-3-3γ at Ser59 and this phosphorylation serves as a switch to dissociate the chaperone from client proteins including LRRK2, a well-established 14-3-3 binding partner. We found that 14-3-3γ phosphorylated by PAK6 is no longer competent to bind LRRK2 at phospho-Ser935, causing LRRK2 dephosphorylation. To address whether these interactions are relevant in a neuronal context, we demonstrate that a constitutively active form of PAK6 rescues the G2019S LRRK2-associated neurite shortening through phosphorylation of 14-3-3γ. Our results identify PAK6 as the kinase for 14-3-3γ and reveal a novel regulatory mechanism of 14-3-3/LRRK2 complex in the brain

Multiple mantle upwellings in the transition zone beneath the northern East-African Rift system from relative P-wave travel-time tomography

Mantle plumes and consequent plate extension have been invoked as the likely cause of East African Rift volcanism. However, the nature of mantle upwelling is debated, with proposed configurations ranging from a single broad plume connected to the large low-shear-velocity province beneath Southern Africa, the so-called African Superplume, to multiple lower-mantle sources along the rift. We present a new P-wave travel-time tomography model below the northern East-African, Red Sea, and Gulf of Aden rifts and surrounding areas. Data are from stations that span an area from Madagascar to Saudi Arabia. The aperture of the integrated data set allows us to image structures of 100 km length-scale down to depths of 700– 800 km beneath the study region. Our images provide evidence of two clusters of low-velocity structures consisting of features with diameter of 100–200 km that extend through the transition zone, the first beneath Afar and a second just west of the Main Ethiopian Rift, a region with off-rift volcanism. Considering seismic sensitivity to temperature, we interpret these features as upwellings with excess temperatures of 100 6 50 K. The scale of the upwellings is smaller than expected for lower mantle plume sources. This, together with the change in pattern of the low-velocity anomalies across the base of the transition zone, suggests that ponding or flow of deep-plume material below the transition zone may be spawning these upper mantle upwellings

GTP binding and intramolecular regulation by the ROC domain of Death Associated Protein Kinase 1

The ROCO proteins are a family of large, multidomain proteins characterised by the presence of a Ras of complex proteins (ROC) domain followed by a COR, or C-terminal of ROC, domain. It has previously been shown that the ROC domain of the human ROCO protein Leucine Rich Repeat Kinase 2 (LRRK2) controls its kinase activity. Here, the ability of the ROC domain of another human ROCO protein, Death Associated Protein Kinase 1 (DAPK1), to bind GTP and control its kinase activity has been evaluated. In contrast to LRRK2, loss of GTP binding by DAPK1 does not result in loss of kinase activity, instead acting to modulate this activity. These data highlight the ROC domain of DAPK1 as a target for modifiers of this proteins function, and casts light on the role of ROC domains as intramolecular regulators in complex proteins with implications for a broad range of human diseases

LRRK2 Biology from structure to dysfunction: research progresses, but the themes remain the same

Since the discovery of leucine-rich repeat kinase 2 (LRRK2) as a protein that is likely central to the aetiology of Parkinson's disease, a considerable amount of work has gone into uncovering its basic cellular function. This effort has led to the implication of LRRK2 in a bewildering range of cell biological processes and pathways, and probable roles in a number of seemingly unrelated medical conditions. In this review we summarise current knowledge of the basic biochemistry and cellular function of LRRK2. Topics covered include the identification of phosphorylation substrates of LRRK2 kinase activity, in particular Rab proteins, and advances in understanding the activation of LRRK2 kinase activity via dimerisation and association with membranes, especially via interaction with Rab29. We also discuss biochemical studies that shed light on the complex LRRK2 GTPase activity, evidence of roles for LRRK2 in a range of cell signalling pathways that are likely cell type specific, and studies linking LRRK2 to the cell biology of organelles. The latter includes the involvement of LRRK2 in autophagy, endocytosis, and processes at the trans-Golgi network, the endoplasmic reticulum and also key microtubule-based cellular structures. We further propose a mechanism linking LRRK2 dimerisation, GTPase function and membrane recruitment with LRRK2 kinase activation by Rab29. Together these data paint a picture of a research field that in many ways is moving forward with great momentum, but in other ways has not changed fundamentally. Many key advances have been made, but very often they seem to lead back to the same places