Multiple supramolecular structures formed by interaction of actin with protamine

When protamine is added to actin, different supramolecular structures are formed depending on the molar ratio of the two proteins and of the ionic strength of the medium. At low ionic strength, and going from a molar ratio of protamine to G-actin of 4:1, 2:1 and 1:1, globular aggregates are first converted into extended structures and then to long threads in which the constituent ATP–G-actin is rapidly exchangeable with the actin of the medium. At high ionic strength {Tyrode [(1910) Arch. Int. Pharmacodyn. Ther. 20, 205–212] solution}, starting from G-actin and protamine in the 1:1 molar ratio, long ropes are formed that can be resolved into intertwining filaments of 4–5nm diameter. The addition of protamine in a 1:1 molar ratio to a solution of F-actin in Tyrode solution causes the breakage of the actin filaments, which is also revealed by the decrease of the viscosity of the solution and the formation of ordered latero-lateral aggregates. The structures formed by reaction of protamine with G-actin can be separated from free G-actin and protamine by filtration through 0.45μm-pore-size Millipore filters. This technique has been exploited to study the exchange reaction between free actin and the actin–protamine complexes. For these studies the 1:1 actin–protamine complex formed at low ionic strength and the 2:1 actin–protamine complex formed in the presence of 23nm-free Mg²⁺ have been selected. In the first case the exchange reaction is practically complete in the dead time of the experiment (20s). In the second case, where the complex operates like a true ATPase, the rate of the exchange is initially comparable with the rate of the ATP cleavage. Later on, however, the complex undergoes a change and the rate of the exchange between free actin and the actin bound to protamine becomes lower than the rate of the ATPase reaction. It is proposed that the ATP exchanges for ADP directly on the G-actin bound in the complex.     

Surfing parameter hyperspaces under climate change scenarios to design future rice ideotypes

Growing food crops to meet global demand and the search for more sustainable cropping systems are increasing the need for new cultivars in key production areas. This study presents the identification of rice traits putatively producing the largest yield benefits in five areas that markedly differ in terms of environmental conditions in the Philippines, India, China, Japan and Italy. The ecophysiological model WARM and sensitivity analysis techniques were used to evaluate phenotypic traits involved with light interception, photosynthetic efficiency, tolerance to abiotic stressors, resistance to fungal pathogens and grain quality. The analysis involved only model parameters that have a close relationship with phenotypic traits breeders are working on, to increase the in vivo feasibility of selected ideotypes. Current climate and future projections were considered, in the light of the resources required by breeding programs and of the role of weather variables in the identification of promising traits. Results suggest that breeding for traits involved with disease resistance, and tolerance to cold‐ and heat‐induced spikelet sterility could provide benefits similar to those obtained from the improvement of traits involved with canopy structure and photosynthetic efficiency. In contrast, potential benefits deriving from improved grain quality traits are restricted by weather variability and markedly affected by G × E interactions. For this reason, district‐specific ideotypes were identified using a new index accounting for both their productivity and feasibility.     

A Cdc7 kinase inhibitor restricts initiation of DNA replication and has antitumor activity

Cdc7 is an essential kinase that promotes DNA replication by activating origins of replication. Here, we characterized the potent Cdc7 inhibitor PHA-767491 (1) in biochemical and cell-based assays, and we tested its antitumor activity in rodents. We found that the compound blocks DNA synthesis and affects the phosphorylation of the replicative DNA helicase at Cdc7-dependent phosphorylation sites. Unlike current DNA synthesis inhibitors, PHA-767491 prevents the activation of replication origins but does not impede replication fork progression, and it does not trigger a sustained DNA damage response. Treatment with PHA-767491 results in apoptotic cell death in multiple cancer cell types and tumor growth inhibition in preclinical cancer models. To our knowledge, PHA-767491 is the first molecule that directly affects the mechanisms controlling initiation as opposed to elongation in DNA replication, and its activities suggest that Cdc7 kinase inhibition could be a new strategy for the development of anticancer therapeutics.     

The interaction of histone and protamine with actin. Possible involvement in the formation of the mitotic spindle

At low ionic strength 1–2 μM protamine or 1–2 μM H1 histone induce the nucleation of G-ATP actin. At high ionic strength 1–2 μM protamine or 0.1 to 0.2 μM H1 histone accelerate by 3 to 4 times the rate of polymerisation of G-ATP actin. It is suggested that histone may trigger the formation of actin fibers running from the kinetochore of the chromosome to the pole of the mitotic spindle.     

Misting and fan cooling of the rest area in a dairy barn

This summer study aimed to evaluate the effect on dairy cows, kept in a free stall barn equipped with fans and sprinklers in the feeding area, of the delivery rate of misters in a cooling system in rest areas with different bedding materials. Thirty cows were divided into two homogenous groups according to milk yield and kept in two pens: one had beds with sand (SAMM) while the other had straw (STLM). Each pen was equipped with 2 fans (Ø 70 cm, 0.50 kW) and 2 misters (delivery rate of 11.2 and 22.5 L/h in STLM and SAMM, respectively) in the rest area. Microclimatic parameters, rectal temperature (RT), breathing rate (BR), milk yield, and some milk traits were recorded. Behavioural routines of the cows (standing and lying) were also continuously recorded during the hotter days. During the trial, two mild–moderate heat waves were observed. During these hotter periods, the daily maximum temperature recorded in the rest areas was 28.9 in SAMM and 31.2 in STLM, and the daily maximum THI was 78.2 in SAMM and 81.5 in STLM. In these periods, the cows in SAMM compared with those in STLM showed lower BR (P?相似文献   

Morpho-biometric and biochemical responses in lettuce seedlings treated by different application methods of Chlorella vulgaris extract: foliar spray or root drench?

Journal of Applied Phycology - Microalgae-based products applied as biostimulants have recently attracted the attention of researchers. The effect of two different methods of application of a...     

Molecular characteristics of chicken liver arginase

A purification procedure for the preparation of chicken liver arginase in a homogeneous form is presented. The enzyme hydrolyses both arginine and argininic acid. Kinetic analysis reveals that the enzyme binds arginine when the amino group is protonated or unprotonated; however, the unprotonated form seems to be hydrolysed more rapidly. The enzyme exchanges with the medium approx. 1.6Mn(2+) ions per molecule.     

Fan cooling of the resting area in a free stalls dairy barn

This summer study evaluated the effect of providing additional fans (cooling) in the resting area within a free-stall dairy barn that had fans and sprinklers in the feeding area and paddock availability. Thirty cows were divided into two homogenous groups and kept in two pens: one had the resting area equipped with two fans (FAN) while no fans were added to the other resting area (CON). Microclimatic parameters, rectal temperature (RT), breathing rate (BR), milk yield, and milk pH traits were recorded. Time budgeting and the behaviour of the cows (time spent in the feeding area, standing and lying in other areas) were also recorded using digital video technology. Two slight-to-moderate heat waves were observed. During the hottest period the daily maximum temperature recorded was 33.5 °C and the daily maximum THI was 81.6. During this period, the BR and RT increased only slightly in both groups, with lower BR (n.s.) in FAN compared with CON. Milk yield was better maintained (n.s.) in FAN compared with CON during the hottest period. The FAN cows showed a greater (P?<?0.05) lying time in the free stalls (9.5 and 8.6 h/day in FAN and CON, respectively), whereas CON cows made greater (P?<?0.05) use of the paddock during evening and late evening hours. Consequently, the total daily lying time was 13.5 h/day in both groups. In conclusion, the results suggest that using fans in the resting area improves cow comfort, which increases use of the resting area. The lying time results also suggest that the benefits of providing ventilation in the resting area might be more evident in barns where there is no paddock.