Global mRNA expression analysis in myosin II deficient strains of Saccharomyces cerevisiae reveals an impairment of cell integrity functions

<p>Abstract</p> <p>Background</p> <p>The <it>Saccharomyces cerevisiae MYO1 </it>gene encodes the myosin II heavy chain (Myo1p), a protein required for normal cytokinesis in budding yeast. Myo1p deficiency in yeast (<it>myo1Δ</it>) causes a cell separation defect characterized by the formation of attached cells, yet it also causes abnormal budding patterns, formation of enlarged and elongated cells, increased osmotic sensitivity, delocalized chitin deposition, increased chitin synthesis, and hypersensitivity to the chitin synthase III inhibitor Nikkomycin Z. To determine how differential expression of genes is related to these diverse cell wall phenotypes, we analyzed the global mRNA expression profile of <it>myo1Δ </it>strains.</p> <p>Results</p> <p>Global mRNA expression profiles of <it>myo1Δ </it>strains and their corresponding wild type controls were obtained by hybridization to yeast oligonucleotide microarrays. Results for selected genes were confirmed by real time RT-PCR. A total of 547 differentially expressed genes (p ≤ 0.01) were identified with 263 up regulated and 284 down regulated genes in the <it>myo1Δ </it>strains. Gene set enrichment analysis revealed the significant over-representation of genes in the protein biosynthesis and stress response categories. The <it>SLT2/MPK1 </it>gene was up regulated in the microarray, and a <it>myo1Δslt2Δ </it>double mutant was non-viable. Overexpression of ribosomal protein genes <it>RPL30 </it>and <it>RPS31 </it>suppressed the hypersensitivity to Nikkomycin Z and increased the levels of phosphorylated Slt2p in <it>myo1Δ </it>strains. Increased levels of phosphorylated Slt2p were also observed in wild type strains under these conditions.</p> <p>Conclusion</p> <p>Following this analysis of global mRNA expression in yeast <it>myo1Δ </it>strains, we conclude that 547 genes were differentially regulated in <it>myo1Δ </it>strains and that the stress response and protein biosynthesis gene categories were coordinately regulated in this mutant. The <it>SLT2/MPK1 </it>gene was confirmed to be essential for <it>myo1Δ </it>strain viability, supporting that the up regulated stress response genes are regulated by the <it>PKC1 </it>cell integrity pathway. Suppression of Nikkomycin Z hypersensitivity together with Slt2p phosphorylation was caused by the overexpression of ribosomal protein genes <it>RPL30 </it>and <it>RPS31</it>. These ribosomal protein mRNAs were down regulated in the <it>myo1Δ </it>arrays, suggesting that down regulation of ribosomal biogenesis may affect cell integrity in <it>myo1Δ </it>strains.</p

Defense techniques for low-rate DoS attacks against application servers

a b s t r a c t Low-rate denial of service (DoS) attacks have recently emerged as new strategies for denying networking services. Such attacks are capable of discovering vulnerabilities in protocols or applications behavior to carry out a DoS with low-rate traffic. In this paper, we focus on a specific attack: the low-rate DoS attack against application servers, and address the task of finding an effective defense against this attack. Different approaches are explored and four alternatives to defeat these attacks are suggested. The techniques proposed are based on modifying the way in which an application server accepts incoming requests. They focus on protective measures aimed at (i) preventing an attacker from capturing all the positions in the incoming queues of applications, and (ii) randomizing the server operation to eliminate possible vulnerabilities due to predictable behaviors. We extensively describe the suggested techniques, discussing the benefits and drawbacks for each under two criteria: the attack efficiency reduction obtained, and the impact on the normal operation of the server. We evaluate the proposed solutions in a both a simulated and a real environment, and provide guidelines for their implementation in a production system

Assessment of the Carbon Stock in Pine Plantations in Southern Spain through ALS Data and K-Nearest Neighbor Algorithm Based Models

Accurate estimation of forest biomass to enable the mapping of forest C stocks over large areas is of considerable interest nowadays. Airborne laser scanning (ALS) systems bring a new perspective to forest inventories and subsequent biomass estimation. The objective of this research was to combine growth models used to update old inventory data to a reference year, low-density ALS data, and k-nearest neighbor (kNN) algorithm Random Forest to conduct biomass inventories aimed at estimating the C sequestration capacity in large Pinus plantations. We obtained a C stock in biomass (Wt-S) of 12.57 Mg·ha−1, ranging significantly from 19.93 Mg·ha−1 for P. halepensis to 49.05 Mg·ha−1 for P. nigra, and a soil organic C stock of the composite soil samples (0–40 cm) ranging from 20.41 Mg·ha−1 in P. sylvestris to 37.32 Mg·ha−1 in P. halepensis. When generalizing these data to the whole area, we obtained an overall C-stock value of 48.01 MgC·ha−1, ranging from 23.96 MgC·ha−1 for P. halepensis to 58.09 MgC·ha−1 for P. nigra. Considering the mean value of the on-site C stock, the study area sustains 1,289,604 Mg per hectare (corresponding to 4,732,869 Mg CO2), with a net increase of 4.79 Mg·ha−1·year−1. Such C cartography can help forest managers to improve forest silviculture with regard to C sequestration and, thus, climate change mitigation

Production of Cellulosic Pulp from Reed (Phragmites australis) to Produce Paper and Paperboard

Due to the global increase in the demand for paper and cardboard, and depletion in some areas of wood resources (main raw material in the paper industry), it has been necessary to investigate new raw materials to substitute the conventional ones. In this work, Phragmites australis, commonly known as reed, has been studied. It has been characterized the chemical composition of the raw material by determining the ash content, α-cellulose, holocellulose, lignin and their extractables in ethanol, 1% soda and hot water. Subsequently three pulping processes have been conducted, using the “soda method" under three different operating conditions. The cellulosic pulps and the papersheets made from the pulps were analysed in order to determine the chemical and physical properties following the relevant TAPPI, UNE and ISO standards. Comparing the results obtained, it can be possible to conclude that the pulp obtained under the following operating conditions: 175°, 90 minutes and 14% of NaOH, presents the best results

Late immune response and chronic infection of vascular graft. A controversial diagnosis

La prevalencia de reacciones periprotésicas estériles en injertos vasculares es de aproximadamente un 0,8%. Son situaciones clínicas donde resulta difícil determinar si su origen reside en una infección por gérmenes de crecimiento lento o es secundario a un rechazo inmunológico al material o por una desestructuración del mismo. Revisamos y comentamos las principales características para distinguir ambas situaciones clínicas. Se aporta un ejemplo clínico que trata de un paciente con injerto vascular de Dacron y una colección de 6 años de evolución, estéril, que rodea el total de la prótesis y requiere su explante y sustitución por un nuevo injerto. En conclusión, ante los casos de reacciones periprotésicas tardías es obligado excluir la presencia de infecciones latentes, pero se recomienda profundizar en el estudio de la respuesta inmunológica frente a los materiales del injerto vascular.The prevalence of sterile perigraft reaction is approximately 0.8% in vascular grafts. To diagnose between an infection by germs of slow growth or a secondary reaction to an immune rejection of the material is difficult in this clinical situation. We review and discuss the main features to distinguish these two clinical possibilities. We report a patient case report with dacron vascular graft and a perigraft collection, sterile, around the total prosthesis which required explantation and replacement by a new graft. In conclusion, to cases of late periprosthetic reactions is required to exclude the presence of latent infections, but recommended further study of the immune response against vascular graft materials

Importancia del peso de semilla en especies del género Quercus. Evidencias en condiciones controladas y experimentos de campo

Congreso celebrado del 21-25 de septiembre 2009, en Ávila, España.Peer reviewe

Maf/ham1-like pyrophosphatases of noncanonical nucleotides are host-specific partners of viral RNA-dependent RNA polymerases.

Cassava brown streak disease (CBSD), dubbed the “Ebola of plants”, is a serious threat to food security in Africa caused by two viruses of the family Potyviridae: cassava brown streak virus (CBSV) and Ugandan (U)CBSV. Intriguingly, U/CBSV, along with another member of this family and one secoviridae, are the only known RNA viruses encoding a protein of the Maf/ham1-like family, a group of widespread pyrophosphatase of non-canonical nucleotides (ITPase) expressed by all living organisms. Despite the socio-economic impact of CDSD, the relevance and role of this atypical viral factor has not been yet established. Here, using an infectious cDNA clone and reverse genetics, we demonstrate that UCBSV requires the ITPase activity for infectivity in cassava, but not in the model plant Nicotiana benthamiana. HPLC-MS/MS experiments showed that, quite likely, this host-specific constraint is due to an unexpected high concentration of non-canonical nucleotides in cassava. Finally, protein analyses and experimental evolution of mutant viruses indicated that keeping a fraction of the yielded UCBSV ITPase covalently bound to the viral RNA-dependent RNA polymerase (RdRP) optimizes viral fitness, and this seems to be a feature shared by the other members of the Potyviridae family expressing Maf/ham1-like proteins. All in all, our work (i) reveals that the over-accumulation of non-canonical nucleotides in the host might have a key role in antiviral defense, and (ii) provides the first example of an RdRP-ITPase partnership, reinforcing the idea that RNA viruses are incredibly versatile at adaptation to different host setups.post-print2435 K

Intrusion Detection With Unsupervised Techniques for Network Management Protocols Over Smart Grids

[Abstract] The present research work focuses on overcoming cybersecurity problems in the Smart Grid. Smart Grids must have feasible data capture and communications infrastructure to be able to manage the huge amounts of data coming from sensors. To ensure the proper operation of next-generation electricity grids, the captured data must be reliable and protected against vulnerabilities and possible attacks. The contribution of this paper to the state of the art lies in the identification of cyberattacks that produce anomalous behaviour in network management protocols. A novel neural projectionist technique (Beta Hebbian Learning, BHL) has been employed to get a general visual representation of the traffic of a network, making it possible to identify any abnormal behaviours and patterns, indicative of a cyberattack. This novel approach has been validated on 3 different datasets, demonstrating the ability of BHL to detect different types of attacks, more effectively than other state-of-the-art methods