Rat muscle acylphosphatase: Purification, amino sequence, and immunological characterization

Acylphosphatase was purified from rat skeletal muscle essentially by gel filtration and high-performance ion-exchange chromatography. The complete amino acid sequence was reconstructed by using the sequence data obtained from tryptic, peptic, andS. aureus V8 protease peptides. The protein consists of 96 amino acid residues and is acetylated at the NH₂-terminus. The immunological cross-reactivity of acylphosphatase from rat and horse skeletal muscle was examined by ELISA. The reaction with rabbit antiserum revealed the presence of at least five antigenic sites on rat enzyme, two of which are common to horse muscle enzyme. Anti-rat antibodies also recognize the peptide that corresponds to the initial part of the molecule, which varies greatly from equine enzyme. Two completely new antigenic sites are herein described: the first can be considered the main antigenic site and is located within positions 21–36, the second is in the COOH-terminal part of the molecule. A mixture of immunoreactive peptides gives strong antibody-antigen reaction inhibition (94%).     

A federated approach to Android malware classification through Perm-Maps

In the last decades, mobile-based apps have been increasingly used in several application fields for many purposes involving a high number of human activities. Unfortunately, in addition to this, the number of cyber-attacks related to mobile platforms is increasing day-by-day. However, although advances in Artificial Intelligence science have allowed addressing many aspects of the problem, malware classification tasks are still challenging. For this reason, the following paper aims to propose new special features, called permission maps (Perm-Maps), which combine information related to the Android permissions and their corresponding severity levels. Such features have proven to be very effective in classifying different malware families through the usage of a convolutional neural network. Also, the advantages introduced by the Perm-Maps have been enhanced by a training process based on a federated logic. Experimental results show that the proposed approach achieves up to a 3% improvement in average accuracy with respect to J48 trees and Naive Bayes classifier, and up to 16% compared to multi-layer perceptron classifier. Furthermore, the combined use of Perm-Maps and federated logic allows dealing with unbalanced training datasets with low computational efforts.

     

Trends of Technical Changes in Rice-Based Farming Systems in Southern China: Case Study of Qianjiang Municipality

This is the second of a series of four articles aimed at providing a general analysis of the dynamic of rural intensification in China in view of sustainability, with a particular focus on the area considered in our project. Data provided in this article confirms the analysis given in the previous article about global trends for agricultural changes. In the last decades, the agricultural sector of the Hubei province managed to cope with the dramatic increase in demographic pressure by increasing more than proportionally agricultural yields. This was paid for in terms of higher environmental stress and higher reliance on external inputs. The challenge for future decades will refer to the need of matching an increasing socioeconomic pressure (=boosting the productivity of labor of farmers to get them a higher income and to make available surplus of food for the growing urban population) while limiting the environmental loading generated by agriculture. Data describing the evolution of cropping patterns, production techniques, mix and intensity of inputs used in production in the last decades seem to indicate that this would represent a formidable challenge. The nature of this challenge implies that the entire Chinese society must be involved in order to negotiate viable compromises between the need of: guaranteeing food security for China; raising the standard of living for rural populations; preserving natural resources and ecological systems for future generations.     

Natural variants suppress mutations in hundreds of essential genes

The consequence of a mutation can be influenced by the context in which it operates. For example, loss of gene function may be tolerated in one genetic background, and lethal in another. The extent to which mutant phenotypes are malleable, the architecture of modifiers and the identities of causal genes remain largely unknown. Here, we measure the fitness effects of ~ 1,100 temperature‐sensitive alleles of yeast essential genes in the context of variation from ten different natural genetic backgrounds and map the modifiers for 19 combinations. Altogether, fitness defects for 149 of the 580 tested genes (26%) could be suppressed by genetic variation in at least one yeast strain. Suppression was generally driven by gain‐of‐function of a single, strong modifier gene, and involved both genes encoding complex or pathway partners suppressing specific temperature‐sensitive alleles, as well as general modifiers altering the effect of many alleles. The emerging frequency of suppression and range of possible mechanisms suggest that a substantial fraction of monogenic diseases could be managed by modulating other gene products.     

Plant regeneration from immature seeds of Eugenia myrtifolia Sims.

Eugenia myrtifolia Sims. is an evergreen shrub, native to temperate and tropical rainforests of Australia, which is becoming an important containerized ornamental plant in the US and Mediterranean nursery industries. To satisfy the growing market demands for this new ornamental plant, development of an accelerated propagation method is required. The goal of this study was to investigate the in vitro regeneration potential of E. myrtifolia Sims. seeds at different stages of development, towards establishment of an in vitro multiplication system. Maximum regeneration of adventitious shoots was achieved from immature seeds cultured in the dark on half-strength Murashige and Skoog (MS) macronutrients and full-strength MS micronutrients and vitamins (MS/2) medium supplemented with 2.5 μM thidiazuron (TDZ). Induction of regeneration occurred after at least two successive subcultures on TDZ-enriched medium, followed by subcultures on expression medium (hormone free MS/2) or multiplication medium [MM; MS medium enriched with 4.4 M 6-benzyladenine and 0.05 M α-naphthaleneacetic acid], where complete development of shoots occurred. The regenerated shoots were excised and transferred again onto MM for micropropagation, where a proliferation rate of 1:4 was achieved, and finally the shoots were transferred to a hormone-free MS medium for rooting. Following ex vitro transplanting, acclimatization over a period of 15 d was sufficient to establish greenhouse plants. The regenerated plants grown in the field for more than 2 yr showed the same phenotype as that of mother plants. The adventitious regeneration and micropropagation carried out in this study can be used for a large-scale propagation and genetic engineering of E. myrtifolia Sims.     

CRISPR gRNA phenotypic screening in zebrafish reveals pro-regenerative genes in spinal cord injury

Zebrafish exhibit robust regeneration following spinal cord injury, promoted by macrophages that control post-injury inflammation. However, the mechanistic basis of how macrophages regulate regeneration is poorly understood. To address this gap in understanding, we conducted a rapid in vivo phenotypic screen for macrophage-related genes that promote regeneration after spinal injury. We used acute injection of synthetic RNA Oligo CRISPR guide RNAs (sCrRNAs) that were pre-screened for high activity in vivo. Pre-screening of over 350 sCrRNAs allowed us to rapidly identify highly active sCrRNAs (up to half, abbreviated as haCRs) and to effectively target 30 potentially macrophage-related genes. Disruption of 10 of these genes impaired axonal regeneration following spinal cord injury. We selected 5 genes for further analysis and generated stable mutants using haCRs. Four of these mutants (tgfb1a, tgfb3, tnfa, sparc) retained the acute haCR phenotype, validating the approach. Mechanistically, tgfb1a haCR-injected and stable mutant zebrafish fail to resolve post-injury inflammation, indicated by prolonged presence of neutrophils and increased levels of il1b expression. Inhibition of Il-1β rescues the impaired axon regeneration in the tgfb1a mutant. Hence, our rapid and scalable screening approach has identified functional regulators of spinal cord regeneration, but can be applied to any biological function of interest.     

Quantitative evaluation of chromosomal rearrangements in gene-edited human stem cells by CAST-Seq

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Immunotoxin mediated killing of clonable T-lymphocytes infiltrating an irreversibly rejected human renal allograft

We isolated and treated in vitro with a novel CD5-specific saporin immunotoxin, referred to as OKT1-SAP, the cells infiltrating an irreversibly rejected renal allograft from a patient who rejected while on cyclosporine plus steroids and then failed to respond to multiple courses of high-dose steroids, intravenous OKT3 antibody, and local irradiation to the graft. We report here that under experimental conditions achievable in vivo the immunotoxin OKT1-SAP was capable of eliminating in vitro more than 95% of clonable T-lymphocytes infiltrating the rejected allograft of this patient despite their resistance to previous aggressive immunosuppression. To our knowledge, this is the first report of an immunotoxin-mediated suppression of the clonogenic growth of rejected renal allograft infiltrating T-lymphocytes.