Deamination of 2',3'-O-isopropylideneadenosine-5'- carboxylic acid catalyzed by adenosine deaminase (ADA) and adenylate deaminase (AMPDA): influence of substrate ionization on the activity of the enzymes

Adenosine deaminase (ADA) and adenylate deaminase (AMPDA) catalyze the deamination of 2 ',3 '-O-isopropylideneadenosine-5'-carboxylic acid to the corresponding inosine derivative and dependence of the rate of enzymatic reaction on the ionization degree of the substrate has been studied at different pH values.     

A drug repurposing strategy for overcoming human multiple myeloma resistance to standard-of-care treatment

Despite several approved therapeutic modalities, multiple myeloma (MM) remains an incurable blood malignancy and only a small fraction of patients achieves prolonged disease control. The common anti-MM treatment targets proteasome with specific inhibitors (PI). The resulting interference with protein degradation is particularly toxic to MM cells as they typically accumulate large amounts of toxic proteins. However, MM cells often acquire resistance to PIs through aberrant expression or mutations of proteasome subunits such as PSMB5, resulting in disease recurrence and further treatment failure. Here we propose CuET—a proteasome-like inhibitor agent that is spontaneously formed in-vivo and in-vitro from the approved alcohol-abuse drug disulfiram (DSF), as a readily available treatment effective against diverse resistant forms of MM. We show that CuET efficiently kills also resistant MM cells adapted to proliferate under exposure to common anti-myeloma drugs such as bortezomib and carfilzomib used as the first-line therapy, as well as to other experimental drugs targeting protein degradation upstream of the proteasome. Furthermore, CuET can overcome also the adaptation mechanism based on reduced proteasome load, another clinically relevant form of treatment resistance. Data obtained from experimental treatment-resistant cellular models of human MM are further corroborated using rather unique advanced cytotoxicity experiments on myeloma and normal blood cells obtained from fresh patient biopsies including newly diagnosed as well as relapsed and treatment-resistant MM. Overall our findings suggest that disulfiram repurposing particularly if combined with copper supplementation may offer a promising and readily available treatment option for patients suffering from relapsed and/or therapy-resistant multiple myeloma.Subject terms: Myeloma, Cancer therapeutic resistance     

Effects of a Copper-Deficient Diet on the Biochemistry,Neural Morphology and Behavior of Aged Mice

Copper dyshomeostasis has been suggested as an aetiological risk factor for some neurodegenerative diseases, such as Alzheimer’s disease. However, the precise mechanism at the base of this involvement is still obscure. In this work, we show the effects of a copper-deficient diet in aged CD1 mice and the influence of such a diet on: a) the concentration of various metal ions (aluminium, copper, iron, calcium, zinc) in the main organs and in different brain areas; b) the alteration of metallothioneins I-II and tyrosine hydroxylase immunopositivity in the brain; c) behavioural tests (open field, pole, predatory aggression, and habituation/dishabituation smell tests). Our data suggested that the copper-deficiency was able to produce a sort of “domino effect” which altered the concentration of the other tested metal ions in the main organs as well as in the brain, without, however, significantly affecting the animal behaviour.     

A kinetic study of phosphorus absorption by excised maize roots from flowing solutions with different phosphorus concentrations

The effect of two different mechanisms of phosphorus ion transport from the nutrient solution volume to the surface areas of excised maize roots was studied under concentrations ranging from 0.01 mM to 50.0 mM KH₂PO₄. A modified technique of study of kinetic ion absorption was used. In the control series, the roots were placed in absorption solution without flow (the dominant mechanism of ion transport to the roots being diffusion), while in the experimental series the absorption solution was flowing round the roots at a rate of 0.162 cm s^?1 (the dominant mechanism of ion transport to the roots being mass flow). The rate of phosphorus absorption by the roots from flowing solutions was highly significantly increased at all concentrations of absorption solution except for the 50.0 mM KH₂PO₄ concentration. The increase in phosphorus absorption in the case of 50.0 mM KH₂PO₄ concentration was non-significant due to the fact that the high concentration of phosphorus together with the diffusion of phosphorus ions ensured a sufficient supply of phosphorus to the roots, covering the requirement for their uptake. The results point to the need for an analysis of environmental factors to be carried out in studying ion absorption kinetics, and reveal the inadequacy of methods usually employed in such investigations, in particular with respect to the homogeneity of the nutrient solution in the whole of its volume and especially round the roots.     

Maize growth and ion absorption in Richter's solution at different flow rates

By comparing maize plants cultivated in standing nutrient solution with those from solutions flowing at different flow rates it has been established that absorption of nitrogen, potassium and especially of phosphorus was increased owing to the flow. There was likewise a relative rise in the distribution of nutrients to the overground parts of the plants. The content expressed per unit dry matter was increased only in the case of phosphorus; with nitrogen and potassium it was slightly lower than in the standing solution. Increasing amounts of iron were required under the conditions of flowing nutrient solutions to prevent chlorosis of the plants. The production of dry matter,NAR andRGR was also increased because of the flow. The flow considerably changed the habitus of the primary roots of the maize plants. The roots were longer, thinner and on the whole they contained relatively less dry matter (RWR). The lengthening of the roots is explained as a response to stimulation by the solution flow—the rheotropism.     

Tailoring the Antibody Response to Aggregated A? Using Novel Alzheimer-Vaccines

Recent evidence suggests Alzheimer-Disease (AD) to be driven by aggregated Aß. Capitalizing on the mechanism of molecular mimicry and applying several selection layers, we screened peptide libraries for moieties inducing antibodies selectively reacting with Aß-aggregates. The technology identified a pool of peptide candidates; two, AFFITOPES AD01 and AD02, were assessed as vaccination antigens and compared to Aβ1-6, the targeted epitope. When conjugated to Keyhole Limpet Hemocyanin (KLH) and adjuvanted with aluminum, all three peptides induced Aß-targeting antibodies (Abs). In contrast to Aß1-6, AD01- or AD02-induced Abs were characterized by selectivity for aggregated forms of Aß and absence of reactivity with related molecules such as Amyloid Precursor Protein (APP)/ secreted APP-alpha (sAPPa). Administration of AFFITOPE-vaccines to APP-transgenic mice was found to reduce their cerebral amyloid burden, the associated neuropathological alterations and to improve their cognitive functions. Thus, the AFFITOME-technology delivers vaccines capable of inducing a distinct Ab response. Their features may be beneficial to AD-patients, a hypothesis currently tested within a phase-II-study.     

Targeted disruption of Mib2 causes exencephaly with a variable penetrance

Mib1 and Mib2 ubiquitin ligases are very similar in their domain construction. They partake in the Notch signaling pathway by ubiquitinating the Notch receptors Delta and Jagged prior to endocytosis. We have created a targeted mutation of Mib2 and show that its phenotype is a variable penetrance, failure to close the cranial neural tube. The penetrance depends on the genetic background but it appears that Mib2 is not completely essential in mouse development.     

Puffing in salivary gland chromosomes of picture-winged Hawaiian Drosophila

A portion of the X chromosome was examined for puffing activity shortly before puparium formation in several species of Hawaiian Drosophila. A large puff was induced by incubating excised salivary glands with ecdysone. In two of the seven species studied, the region in question has been moved by inversion to a new position relative to the banding sequence in the other species, but the reaction to ecdysone remained unchanged. Another puff, not affected by short term incubations with ecdysone, displayed different degrees of activity in some homosequential species, affording a cytological means for distinguishing these species.     

Water- and alkali-soluble glucans from oak lichen

A water-soluble glucan, [α]²_D +217° (water), and an alkali-soluble glucan,

+152° (sodium hydroxide), have been isolated from the oak lichen Evernia prunastri (L.) Ach. On the basis of methylation analysis, periodate oxidation, and partial acid hydrolysis, the water-soluble polysaccharide has been shown to be a neutral, slightly branched glucan with a main chain composed of (1→3)- and (1→4)- linked glucopyranose residues in the ratio 1?:1. Branching occurs most probably at position 2 of (1→4)-linked glucopyranose residues. On the basis of optical rotation and i.r. spectral data, and enzymic hydrolysis, the α-D configuration has been assigned to the glycosidic linkages. Likewise, the alkali-soluble polysaccharide was shown to be a neutral, branched glucan with a main chain composed of (1→3)- and (1→4)-linked α-D-glucopyranose residues in the ratio 6:1. Each of the (1→4)-linked units was a branch point involving position 6. The presence of some β-D linkages is not excluded since hydrolysis with β-D-glucosidase occurred to a small extent.