Whole-exome sequencing analysis in twin sibling males with an anterior cruciate ligament rupture.

Familial predisposition is among the major genetic risk factors for non-contact musculoskeletal tissue injuries. Personal genome sequence shows that different polymorphism profiles may account for the number and the degree of injuries and the recovery time. Genotyping studies allow investigation into genome factors with potential impact on pathogenesis of non-contact ligament injuries. We have studied a family with twin sibling males surgically diagnosed of an anterior cruciate ligament non-contact rupture and non-affected progenitors (father and mother) were subjected to whole exome sequencing (WES) analysis. WES analysis previously carried out on 16 individuals, without ACL injury medical records, were also included in this study for single nucleotide variants (SNVs) and small insertions and deletions detection (indels), variant filtering and to prioritize variants relative to the disease. WES analysis to identify SNVs and indels was performed using open web-based bioinformatics tools. A set of 11 new variants shared by family members can be associated to ACL non-contact injury, including SerpinA11, ARSI, NOCHT4, EPB41, FDFT1, POMC, KIF26A, OLFML2B, ATG7, FAH and WDR6. All of them, except ATG7 and WDR6, have shown a damaging predictive pattern by combinatorial standard predictive scores. In combination to the identified SNVs of EPB41 and SerpinA11 genes, ACTL7A gene showed a predicted deleterious variant reinforcing the idea these variants impact on of fibroblast-like cells deformability and ECM misbalance, Differential gene expression and RNA sequencing analysis will help to understand the combined participation of these protein coding genes in ACL non-contact injuries.

Understanding the pathogenesis of hip fracture in the elderly, osteoporotic theory is not reflected in the outcome of prevention programmes.

Hip fractures are an acute and worsening public health problem. They mainly affect elderly people, a population group that is highly vulnerable to disease and accidents, and to falls in particular. Although it has been suggested that osteoporosis is the cause of hip fractures, they mainly occur after a fall has been suffered. The underlying causes of a fall are not related to osteoporosis, although pharmaceutical companies have coined the term "osteoporotic fracture" for hip fractures in the elderly. Drug treatments for osteoporosis have not diminished the frequency of these injuries, nor have they prevented the occurrence of a subsequent fracture. Since pharmaceutical interests require osteoporosis to be considered a disease, rather than a normal condition of senescence, they go further by assuming that treatment for osteoporosis is essential, and that this policy will diminish the incidence of hip fractures. On the other hand, the origin and treatment of conditions that may be conducive to provoking falls are very difficult to elucidate. In this paper, we consider some of the medical and social problems that arise in this area, as well as conflicts of interest regarding the aetiopathogenesis and prevention of hip fracture, and propose a new paradigm for the prevention of falls.

Effectiveness of an individualized program of muscular strength and endurance with aerobic training for improving germ cell cancer-related fatigue in men undergoing chemotherapy: EFICATEST study protocol for a randomized controlled trial.

BACKGROUND: Patients with testicular germ cell cancer (GCC) have a high cure rate; however, cancer-related fatigue is the most common complication among patients with GCC undergoing treatment with chemotherapy. Although exercise is widely recommended, information about the physio-pathological effects of cancer therapy on skeletal muscle is very limited. Our aim is to evaluate the effects of an individualized program of muscular strength and endurance with aerobic training on cancer-related fatigue. METHODS/DESIGN: The present study is a randomized controlled trial comparing an individualized program of muscular strength and endurance with aerobic training compared to a control group. We will conduct this trial in patients undergoing chemotherapy, recruited by the Department of Oncology of Virgen de la Victoria Hospital (Málaga). Patients will be included and evaluated before the first cycle of chemotherapy and assigned randomly to the experimental or control group. Cancer-related fatigue, physical condition and biological samples will be measured at the beginning and at the end of an 8-week intervention by the same evaluator, who will be unaware of the allocation of participants to each group. Furthermore, there will be monitoring for 6 months (24 weeks) after training for all outcome variables. DISCUSSION: This study hopes to offer patients with GCC an individualized exercise program with aerobic training for cancer-related fatigue. Such a scheme, if beneficial, could be implemented successfully within public health. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02433197 . Date of registration: 13 April 2015.

C2 from Beet curly top virus meddles with the cell cycle: a novel function for an old pathogenicity factor.

Geminiviruses are ssDNA plant viruses that infect a wide range of crops. Since geminiviruses often infect terminally differentiated cells, they must induce cell cycle re-entry in order to replicate; until recently, only two viral proteins, the replication-associated protein Rep and the curtoviral pathogenicity factor C4, had been assigned a role in the restoration of cell competency. In a recent work, we demonstrated that C2 from Beet curly top virus activates the expression of host genes involved in DNA replication and/or control of the G2/M transition in a manner consistent with cell cycle re-entry. As expected, expression of BCTV C2 results in enhanced replication of DNA viruses. We conclude that BCTV C2 acts as a re-activator of the cell cycle in infected cells, enhancing the DNA replication competency and providing a cell environment favorable for replication of geminiviruses. Potential mechanisms for this novel function are discussed in light of our findings.

C2 from Beet curly top virus promotes a cell environment suitable for efficient replication of geminiviruses, providing a novel mechanism of viral synergism.

• Geminiviruses are plant viruses with circular, single-stranded (ss) DNA genomes that infect a wide range of species and cause important losses in agriculture. Geminiviruses do not encode their own DNA polymerase, and rely on the host cell machinery for their replication. • Here, we identify a positive effect of the curtovirus Beet curly top virus (BCTV) on the begomovirus Tomato yellow leaf curl Sardinia virus (TYLCSV) infection in Nicotiana benthamiana plants. • Our results show that this positive effect is caused by the promotion of TYLCSV replication by BCTV C2. Transcriptomic analyses of plants expressing C2 unveil an up-regulation of cell cycle-related genes induced on cell cycle re-entry; experiments with two mutated versions of C2 indicate that this function resides in the N-terminal part of C2, which is also sufficient to enhance geminiviral replication. Moreover, C2 expression promotes the replication of other geminiviral species, but not of RNA viruses. • We conclude that BCTV C2 has a novel function in the promotion of viral replication, probably by restoring the DNA replication competency of the infected cells and thus creating a favourable cell environment for viral spread. Because C2 seems to have a broad impact on the replication of geminiviruses, this mechanism might have important epidemiological implications.

Fusarium oxysporum gas1 encodes a putative beta-1,3-glucanosyltransferase required for virulence on tomato plants.

Glycosylphosphatidylinositol-anchored (beta)-1,3-glucanosyltransferases play active roles in fungal cell wall biosynthesis and morphogenesis and have been implicated in virulence on mammals. The role of beta-1,3-glucanosyltransferases in pathogenesis to plants has not been explored so far. Here, we report the cloning and mutational analysis of the gas1 gene encoding a putative beta-1,3-glucanosyltransferase from the vascular wilt fungus Fusarium oxysporum. In contrast to Candida albicans, expression of gas1 in F. oxysporum was independent of ambient pH and of the pH response transcription factor PacC. Gene knockout mutants lacking a functional gas1 allele grew in a way similar to the wildtype strain in submerged culture but exhibited restricted colony growth on solid substrates. The restricted growth phenotype was relieved by the osmotic stabilizer sorbitol, indicating that it may be related to structural alterations in the cell wall. Consistent with this hypothesis, deltagas1 mutants exhibited enhanced resistance to cell wall-degrading enzymes and increased transcript levels of chsV and rho1, encoding a class V chitin synthase and a small monomeric G protein, respectively. The deltagas1 mutants showed dramatically reduced virulence on tomato, both in a root infection assay and in a fruit tissue-invasion model, thus providing the first evidence for an essential role of fungal beta-1,3-glucanosyltransferases during plant infection.

On the role of Trk1 and Trk2 in Schizosaccharomyces pombe under different ion stress conditions.

Trk1 and Trk2 are the major K(+) transport systems in Schizosaccharomyces pombe. Both transporters individually seem to be able to cope with K(+) requirements of the cells under normal conditions, since only the double mutant shows defective K(+) transport and defective growth at limiting K(+) concentrations. We have studied in detail the role of SpTrk1 and SpTrk2 under different ion stress conditions. Results show that the strain with only Trk1 (trk1(+)) is less sensitive to Li(+) and to hygromycin B, it grows better at low K(+) and it survives longer in a medium without K(+) than the strain expressing only Trk2 (trk2(+)). We conclude that Trk1 contributes more efficiently than Trk2 to the performance of the fission yeast under ion stress conditions. In the wild type both trk1(+) and trk2(+) genes are expressed and probably collaborate for the performance of the cells.

Fusarium oxysporum as a multihost model for the genetic dissection of fungal virulence in plants and mammals.

Fungal pathogens cause disease in plant and animal hosts. The extent to which infection mechanisms are conserved between both classes of hosts is unknown. We present a dual plant-animal infection system based on a single strain of Fusarium oxysporum, the causal agent of vascular wilt disease in plants and an emerging opportunistic human pathogen. Injection of microconidia of a well-characterized tomato pathogenic isolate (isolate 4287) into the lateral tail vein of immunodepressed mice resulted in disseminated infection of multiple organs and death of the animals. Knockout mutants in genes encoding a mitogen-activated protein kinase, a pH response transcription factor, or a class V chitin synthase previously shown to be implicated in virulence on tomato plants were tested in the mouse model. The results indicate that some of these virulence factors play functionally distinct roles during the infection of tomato plants and mice. Thus, a single F. oxysporum strain can be used to study fungal virulence mechanisms in plant and mammalian pathogenesis.

pH response transcription factor PacC controls salt stress tolerance and expression of the P-Type Na+ -ATPase Ena1 in Fusarium oxysporum.

Fungi possess efficient mechanisms of pH and ion homeostasis, allowing them to grow over a wide range of environmental conditions. In this study, we addressed the role of the pH response transcription factor PacC in salt tolerance of the vascular wilt pathogen Fusarium oxysporum. Loss-of-function pacC(+/-) mutants showed increased sensitivity to Li(+) and Na(+) and accumulated higher levels of these cations than the wild type. In contrast, strains expressing a dominant activating pacC(c) allele were more salt tolerant and had lower intracellular Li(+) and Na(+) concentrations. Although the kinetics of Li(+) influx were not altered by mutations in pacC, we found that Li(+) efflux at an alkaline, but not at an acidic, ambient pH was significantly reduced in pacC(+/-) loss-of-function mutants. To explore the presence of a PacC-dependent efflux mechanism in F. oxysporum, we cloned ena1 encoding an orthologue of the yeast P-type Na(+)-ATPase ENA1. Northern analysis revealed that efficient transcriptional activation of ena1 in F. oxysporum required the presence of high Na(+) concentrations and alkaline ambient pH and was dependent on PacC function. We propose a model in which PacC controls ion homeostasis in F. oxysporum at a high pH by activating expression of ena1 coordinately with a second Na(+)-responsive signaling pathway.

The pH signalling transcription factor PacC controls virulence in the plant pathogen Fusarium oxysporum.

Gene expression in fungi by ambient pH is regulated via a conserved signalling cascade whose terminal component is the zinc finger transcription factor PacC/Rim1p. We have identified a pacC orthologue in the vascular wilt pathogen Fusarium oxysporum that binds the consensus 5'-GCCAAG-3' sequence and is proteolytically processed in a similar way to PacC from Aspergillus nidulans. pacC transcript levels were elevated in F. oxysporum grown in alkaline conditions and almost undetectable at extreme acidic growth conditions. PacC+/- loss-of-function mutants displayed an acidity-mimicking phenotype resulting in poor growth at alkaline pH, increased acid protease activity and higher transcript levels of acid-expressed polygalacturonase genes. Reintroduction of a functional pacC copy into a pacC+/- mutant restored the wild-type phenotype. Conversely, F. oxysporum merodiploids carrying a dominant activating pacCc allele had increased pacC transcript and protein levels and displayed an alkalinity-mimicking phenotype with reduced acid phosphatase and increased alkaline protease activities. PacC+/- mutants were more virulent than the wild-type strain in root infection assays with tomato plants, whereas pacCc strains were significantly reduced in virulence. We propose that F. oxysporum PacC acts as a negative regulator of virulence to plants, possibly by preventing transcription of acid-expressed genes important for infection.

Fusarium oxysporum: exploring the molecular arsenal of a vascular wilt fungus.

UNLABELLED: SUMMARY Taxonomy: Vascular wilt fungus; Ascomycete although sexual stage is yet to be found. The most closely related teleomorphic group, Gibberella, is classified within the Pyrenomycetes. HOST RANGE: Very broad at the species level. More than 120 different formae speciales have been identified based on specificity to host species belonging to a wide range of plant families. Disease symptoms: Initial symptoms of vascular wilt include vein clearing and leaf epinasty, followed by stunting, yellowing of the lower leafs, progressive wilting of leaves and stem, defoliation and finally death of the plant. In cross-sections of the stem, a brown ring is evident in the area of the vascular bundles. Some formae speciales are not primarily vascular pathogens but cause foot- and rootrot or bulbrot. Economic importance: Causes severe losses on most vegetables and flowers, several field crops such as cotton and tobacco, plantation crops such as banana, plantain, coffee and sugarcane, and a few shade trees. CONTROL: Use of resistant varieties is the only practical measure for controlling the disease in the field. Under greenhouse conditions, soil sterilization can be performed. Alternative control methods with potential for the future include soil solarization and biological control with antagonistic bacteria or fungi. USEFUL WEBSITES: http://www.fgsc.net/fus.htm, http://www-genome.wi.mit.edu/annotation/fungi/fusarium/, http://www.cbs.knaw.nl/fusarium/database.html.