Single-Tear Proteomics: A Feasible Approach to Precision Medicine

Lacrimal fluid is an attractive source of noninvasive biomarkers, the main limitation being the small sample amounts typically collected. Advanced analytical methods to allow for proteomics profiling from a few microliters are needed to develop innovative biomarkers, with attractive perspectives of applications to precision medicine. This work describes an effective, analytical pipeline for single-tear analysis by ultrahigh-resolution, shotgun proteomics from 23 healthy human volunteers, leading to high-confidence identification of a total of 890 proteins. Highly reproducible quantification was achieved by either peak intensity, peak area, or spectral counting. Hierarchical clustering revealed a stratification of females vs. males that did not emerge from previous studies on pooled samples. Two subjects were monitored weekly over 3 weeks. The samples clustered by withdrawal time of day (morning vs. afternoon) but not by follow-up week, with elevated levels of components of the immune system in the morning samples. This study demonstrates feasibility of single-tear quantitative proteomics, envisaging contributions of this unconventional body fluid to individualized approaches in biomedicine.     

Natural co-occurrence of fumonisins, deoxynivalenol, zearalenone and aflatoxins in field trial corn in Argentina

Corn samples collected from the main production area in Argentina in 1995 were surveyed for the natural occurrence of Fusarium mycotoxins and aflatoxins. Fumonisins B₁, B₂ and B₃ and zearalenone were found in all samples. A positive relationship was found between fumonisins B₁, B₂ and B₃, B₁ and B₃, and B₂ and B . Deoxynivalenol and aflatoxins were not de3 tected. Mycological survey has also revealed the predominance of Fusarium moniliforme . This is the first report on the simultaneous occurrence of fumonisins and zearalenone in corn from the main production area in Argentina.     

Developing GIS Applications with Objects: A Design Patterns Approach

In this paper we present an object-oriented approach for designing GIS applications; it combines well known software engineering practices with the use of design patterns as a conceptual tool to cope with recurrent problems appearing in the GIS domain. Our approach allows the designer to decouple the conceptual definition of application objects from their spatial representation. In this way, GIS applications can evolve smoothly, because maintenance is achieved by focusing on different concerns at different times. We show that our approach is also useful to support spatial features in conventional applications built with object-oriented technology. The structure of this paper is as follows: We first introduce design patterns, an efficient strategy to record design experience; then we discuss the most common design problems a developer of GIS applications must face. The core of our method is then presented by explaining how the use of decorators helps in extending objects to incorporate spatial attributes and behavior. Next, we analyze some recurrent design problems in the GIS domain and present some new patterns addressing those problems. Some further work is finally discussed.     

Study on Error Compensation Method of Online Roll Profile Measurement

Flatness and surface quality are important quality indexes of strip steel, and roll wear will directly affect the product's flatness and surface quality. Online roll profile measurement technology can obtain the information of roll profile in real time, guide the optimization of roll change rhythm, and improve the flatness control ability of rolling mill and surface quality of strip product. However, the error caused by roll displacement, clearance, roll thermal crown, and other factors is usually the same order of magnitude as the roll shape or roll wear, or even one order of magnitude higher. So error compensation is the key to ensuring the accuracy of online roll profile measurement. Herein, the position measurement technology of the roll bearing block is considered in the online roll profile measurement, and the measurement error caused by the deviation of the roll axis is separated. The thermal crown model is established to separate the measurement errors caused by the thermal crown. The error compensation model of online roll profile measurement is established, and the simulation calculation is carried out. This error compensation method is of great significance to improve the accuracy of online roll profile measurement and realize the fine management of mill roll in service.     

Contribution of Metabolomics to Multiple Sclerosis Diagnosis,Prognosis and Treatment

Metabolomics-based technologies map in vivo biochemical changes that may be used as early indicators of pathological abnormalities prior to the development of clinical symptoms in neurological conditions. Metabolomics may also reveal biochemical pathways implicated in tissue dysfunction and damage and thus assist in the development of novel targeted therapeutics for neuroinflammation and neurodegeneration. Metabolomics holds promise as a non-invasive, high-throughput and cost-effective tool for early diagnosis, follow-up and monitoring of treatment response in multiple sclerosis (MS), in combination with clinical and imaging measures. In this review, we offer evidence in support of the potential of metabolomics as a biomarker and drug discovery tool in MS. We also use pathway analysis of metabolites that are described as potential biomarkers in the literature of MS biofluids to identify the most promising molecules and upstream regulators, and show novel, still unexplored metabolic pathways, whose investigation may open novel avenues of research.     

The Hedgehog-GLI Pathway Regulates MEK5-ERK5 Expression and Activation in Melanoma Cells

Malignant melanoma is the deadliest skin cancer, with a poor prognosis in advanced stages. We recently showed that the extracellular signal-regulated kinase 5 (ERK5), encoded by the MAPK7 gene, plays a pivotal role in melanoma by regulating cell functions necessary for tumour development, such as proliferation. Hedgehog-GLI signalling is constitutively active in melanoma and is required for proliferation. However, no data are available in literature about a possible interplay between Hedgehog-GLI and ERK5 pathways. Here, we show that hyperactivation of the Hedgehog-GLI pathway by genetic inhibition of the negative regulator Patched 1 increases the amount of ERK5 mRNA and protein. Chromatin immunoprecipitation showed that GLI1, the major downstream effector of Hedgehog-GLI signalling, binds to a functional non-canonical GLI consensus sequence at the MAPK7 promoter. Furthermore, we found that ERK5 is required for Hedgehog-GLI-dependent melanoma cell proliferation, and that the combination of GLI and ERK5 inhibitors is more effective than single treatments in reducing cell viability and colony formation ability in melanoma cells. Together, these findings led to the identification of a novel Hedgehog-GLI-ERK5 axis that regulates melanoma cell growth, and shed light on new functions of ERK5, paving the way for new therapeutic options in melanoma and other neoplasms with active Hedgehog-GLI and ERK5 pathways.     

Hybrid Self-Assembling Nanoparticles Encapsulating Zoledronic Acid: A Strategy for Fostering Their Clinical Use

Self-assembling nanoparticles (SANPs) promise an effective delivery of bisphosphonates or microRNAs in the treatment of glioblastoma (GBM) and are obtained through the sequential mixing of four components immediately before use. The self-assembling approach facilitates technology transfer, but the complexity of the SANP preparation protocol raises significant concerns in the clinical setting due to the high risk of human errors during the procedure. In this work, it was hypothesized that the SANP preparation protocol could be simplified by using freeze-dried formulations. An in-depth thermodynamic study was conducted on solutions of different cryoprotectants, namely sucrose, mannitol and trehalose, to test their ability to stabilize the produced SANPs. In addition, the ability of SANPs to deliver drugs after lyophilization was assessed on selected formulations encapsulating zoledronic acid in vitro in the T98G GBM cell line and in vivo in an orthotopic mouse model. Results showed that, after lyophilization optimization, freeze-dried SANPs encapsulating zoledronic acid could retain their delivery ability, showing a significant inhibition of T98G cell growth both in vitro and in vivo. Overall, these results suggest that freeze-drying may help boost the industrial development of SANPs for the delivery of drugs to the brain.     

Gold Nanoparticles Contact with Cancer Cell: A Brief Update

The fine-tuning of the physicochemical properties of gold nanoparticles has facilitated the rapid development of multifunctional gold-based nanomaterials with diagnostic, therapeutic, and therapeutic applications. Work on gold nanoparticles is increasingly focusing on their cancer application. This review provides a summary of the main biological effects exerted by gold nanoparticles on cancer cells and highlights some critical factors involved in the interaction process (protein corona, tumor microenvironment, surface functionalization). The review also contains a brief discussion of the application of gold nanoparticles in target discovery.     

LAM Cells as Potential Drivers of Senescence in Lymphangioleiomyomatosis Microenvironment

Senescence is a stress-response process characterized by the irreversible inhibition of cell proliferation, associated to the acquisition of a senescence-associated secretory phenotype (SASP), that may drive pathological conditions. Lymphangioleiomyomatosis (LAM) is a rare disease in which LAM cells, featuring the hyperactivation of the mammalian Target of Rapamycin Complex 1 (mTORC1) for the absence of tuberin expression, cause the disruption of the lung parenchyma. Considering that LAM cells secrete SASP factors and that mTOR is also a driver of senescence, we deepened the contribution of senescence in LAM cell phenotype. We firstly demonstrated that human primary tuberin-deficient LAM cells (LAM/TSC cells) have senescent features depending on mTOR hyperactivation, since their high positivity to SA-β galactosidase and to phospho-histone H2A.X are reduced by inducing tuberin expression and by inhibiting mTOR with rapamycin. Then, we demonstrated the capability of LAM/TSC cells to induce senescence. Indeed, primary lung fibroblasts (PLFs) grown in LAM/TSC conditioned medium increased the positivity to SA-β galactosidase and to phospho-histone H2A.X, as well as p21^WAF1/CIP1 expression, and enhanced the mRNA expression and the secretion of the SASP component IL-8. Taken together, these data make senescence a novel field of study to understand LAM development and progression.