Biochemistry and functional aspects of human glandular kallikreins

Human urinary kallikrein was purified by gel filtration on Sephacryl S-200 and affinity chromatography on aprotinin-Sepharose, followed by ion exchange chromatography on DEAE-Sepharose. In dodecylsulfate gel electrophoresis two protein bands with molecular weights of 41,000 and 34,000 were separated. The amino acid composition and the carbohydrate content of the kallikrein preparation were determined; isoleucine was identified as the only aminoterminal amino acid. The bimolecular velocity constant for the inhibition by diisopropyl fluorophosphate was determined as 9±2 l mol–1 min–1. The hydrolysis of a number of substrates was investigated and AcPheArgOEt was found to be the most sensitive substrate for human urinary kallikrein. Using this substrate an assay method for kallikrein in human urine was developed. It was shown by radioimmunoassay that pig pancreatic kallikrein can be absorbed in the rat intestinal tract. Furthermore, in dogs the renal excretion of glandular kallikrein from blood was demonstrated by radioimmunological methods

The ROCK inhibitor Fasudil prevents chronic restraint stress-induced depressive-like behaviors and dendritic spine loss in rat hippocampus

Indexación: Web of Science; Scopus.Background: Dendritic arbor simplification and dendritic spine loss in the hippocampus, a limbic structure implicated in mood disorders, are assumed to contribute to symptoms of depression. These morphological changes imply modifications in dendritic cytoskeleton. Rho GTPases are regulators of actin dynamics through their effector Rho kinase. We have reported that chronic stress promotes depressive-like behaviors in rats along with dendritic spine loss in apical dendrites of hippocampal pyramidal neurons, changes associated with Rho kinase activation. The present study proposes that the Rho kinase inhibitor Fasudil may prevent the stress-induced behavior and dendritic spine loss. Methods: Adult male Sprague-Dawley rats were injected with saline or Fasudil (i.p., 10 mg/kg) starting 4 days prior to and maintained during the restraint stress procedure (2.5 h/d for 14 days). Nonstressed control animals were injected with saline or Fasudil for 18 days. At 24 hours after treatment, forced swimming test, Golgi-staining, and immuno-western blot were performed. Results: Fasudil prevented stress-induced immobility observed in the forced swimming test. On the other hand, Fasudiltreated control animals showed behavioral patterns similar to those of saline-treated controls. Furthermore, we observed that stress induced an increase in the phosphorylation of MYPT1 in the hippocampus, an exclusive target of Rho kinase. This change was accompanied by dendritic spine loss of apical dendrites of pyramidal hippocampal neurons. Interestingly, increased pMYPT1 levels and spine loss were both prevented by Fasudil administration. Conclusion: Our findings suggest that Fasudil may prevent the development of abnormal behavior and spine loss induced by chronic stress by blocking Rho kinase activity.https://academic.oup.com/ijnp/article/20/4/336/263217

Invasive Pathogen Threatens Bird-Pine Mutualism: Implications for Sustaining a High-Elevation Ecosystem

Human-caused disruptions to seed-dispersal mutualisms increase the extinction risk for both plant and animal species. Large-seeded plants can be particularly vulnerable due to highly specialized dispersal systems and no compensatory regeneration mechanisms. Whitebark pine ( Pinus albicaulis), a keystone subalpine species, obligately depends upon the Clark\u27s Nutcracker (Nucifraga columbiana) for dispersal of its large, wingless seeds. Clark\u27s Nutcracker, a facultative mutualist with whitebark pine, is sensitive to rates of energy gain, and emigrates from subalpine forests during periods of cone shortages. The invasive fungal pathogen Cronartium ribicola, which causes white pine blister rust, reduces whitebark pine cone production by killing cone-bearing branches and trees. Mortality from blister rust reaches 90% or higher in some whitebark pine forests in the Northern Rocky Mountains, USA, and the rust now occurs nearly rangewide in whitebark pine. Our objectives were to identify the minimum level of cone production necessary to elicit seed dispersal by nutcrackers and to determine how cone production is influenced by forest structure and health. We quantified forest conditions and ecological interactions between nutcrackers and whitebark pine in three Rocky Mountain ecosystems that differ in levels of rust infection and mortality. Both the frequency of nutcracker occurrence and probability of seed dispersal were strongly related to annual whitebark pine cone production, which had a positive linear association with live whitebark pine basal area, and negative linear association with whitebark pine tree mortality and rust infection. From our data, we estimated that a threshold level of similar to 1000 cones/ha is needed for a high likelihood of seed dispersal by nutcrackers ( probability \u3e= 0.7), and that this level of cone production can be met by forests with live whitebark pine basal area \u3e5.0 m(2)/ha. The risk of mutualism disruption is greatest in northernmost Montana ( USA), where three-year mean cone production and live basal area fell below predicted threshold levels. There, nutcracker occurrence, seed dispersal, and whitebark pine regeneration were the lowest of the three ecosystems. Managers can use these threshold values to differentiate between restoration sites requiring planting of rust-resistant seedlings and sites where nutcracker seed dispersal can be expected

Di- and Trinuclear Mixed-Valence Copper Amidinate Complexes from Reduction of Iodine

Molecular examples of mixed-valence copper complexes through chemical oxidation are rare but invoked in the mechanism of substrate activation, especially oxygen, in copper-containing enzymes. To examine the cooperative chemistry between two metals in close proximity to each other we began studying the reactivity of a dinuclear Cu(I) amidinate complex. The reaction of [(2,6-Me2C6H3N)2C(H)]2Cu2, 1, with I2 in tetrahydrofuran (THF), CH3CN, and toluene affords three new mixed-valence copper complexes [(2,6-Me2C6H3N)2C(H)]2Cu2(μ2-I3)(THF)2, 2, [(2,6-Me2C6H3N)2C(H)]2Cu2(μ2-I) (NCMe)2, 3, and [(2,6-Me2C6H3N)2C(H)]3Cu3(μ3-I)2, 4, respectively. The first two compounds were characterized by UV-vis and electron paramagnetic resonance spectroscopies, and their molecular structure was determined by X-ray crystallography. Both di- and trinuclear mixed-valence intermediates were characterized for the reaction of compound 1 to compound 4, and the molecular structure of 4 was determined by X-ray crystallography. The electronic structure of each of these complexes was also investigated using density functional theory

Study of the time and space distribution of beta+ emitters from 80 MeV/u carbon ion beam irradiation on PMMA

Proton and carbon ion therapy is an emerging technique used for the treatment of solid cancers. The monitoring of the dose delivered during such treatments and the on-line knowledge of the Bragg peak position is still a matter of research. A possible technique exploits the collinear 511\ \kilo\electronvolt photons produced by positrons annihilation from $\beta^+$ emitters created by the beam. This paper reports rate measurements of the 511\ \kilo\electronvolt photons emitted after the interactions of a 80\ \mega\electronvolt / u fully stripped carbon ion beam at the Laboratori Nazionali del Sud (LNS) of INFN, with a Poly-methyl methacrylate target. The time evolution of the $\beta^+$ rate was parametrized and the dominance of $^{11}C$ emitters over the other species ($^{13}N$, $^{15}O$, $^{14}O$) was observed, measuring the fraction of carbon ions activating $\beta^+$ emitters $A_0=(10.3\pm0.7)\cdot10^{-3}$. The average depth in the PMMA of the positron annihilation from $\beta^+$ emitters was also measured, D_{\beta^+}=5.3\pm1.1\ \milli\meter, to be compared to the expected Bragg peak depth D_{Bragg}=11.0\pm 0.5\ \milli\meter obtained from simulations

A measure of centrality based on the spectrum of the Laplacian

We introduce a family of new centralities, the k-spectral centralities. k-Spectral centrality is a measurement of importance with respect to the deformation of the graph Laplacian associated with the graph. Due to this connection, k-spectral centralities have various interpretations in terms of spectrally determined information. We explore this centrality in the context of several examples. While for sparse unweighted networks 1-spectral centrality behaves similarly to other standard centralities, for dense weighted networks they show different properties. In summary, the k-spectral centralities provide a novel and useful measurement of relevance (for single network elements as well as whole subnetworks) distinct from other known measures.Comment: 12 pages, 6 figures, 2 table

Analysis of weighted networks

The connections in many networks are not merely binary entities, either present or not, but have associated weights that record their strengths relative to one another. Recent studies of networks have, by and large, steered clear of such weighted networks, which are often perceived as being harder to analyze than their unweighted counterparts. Here we point out that weighted networks can in many cases be analyzed using a simple mapping from a weighted network to an unweighted multigraph, allowing us to apply standard techniques for unweighted graphs to weighted ones as well. We give a number of examples of the method, including an algorithm for detecting community structure in weighted networks and a new and simple proof of the max-flow/min-cut theorem.Comment: 9 pages, 3 figure