Differentiated cell behavior: a multiscale approach using measure theory

This paper deals with the derivation of a collective model of cell populations out of an individual-based description of the underlying physical particle system. By looking at the spatial distribution of cells in terms of time-evolving measures, rather than at individual cell paths, we obtain an ensemble representation stemming from the phenomenological behavior of the single component cells. In particular, as a key advantage of our approach, the scale of representation of the system, i.e., microscopic/discrete vs. macroscopic/continuous, can be chosen a posteriori according only to the spatial structure given to the aforesaid measures. The paper focuses in particular on the use of different scales based on the specific functions performed by cells. A two-population hybrid system is considered, where cells with a specialized/differentiated phenotype are treated as a discrete population of point masses while unspecialized/undifferentiated cell aggregates are represented with a continuous approximation. Numerical simulations and analytical investigations emphasize the role of some biologically relevant parameters in determining the specific evolution of such a hybrid cell system.Comment: 25 pages, 6 figure

A GFP-based assay for rapid screening of compounds affecting ARE-dependent mRNA turnover

A reporter transcript containing the green fluorescent protein (GFP) gene upstream of the destabilizing 3′-untranslated region (3′-UTR) of the murine IL-3 gene was inserted in mouse PB-3c-15 mast cells. The GFP-IL-3 transcript was inherently unstable due to the presence of an adenosine-uridine (AU)-rich element (ARE) in the 3′-UTR and was subject to rapid decay giving a low baseline of GFP fluorescence. Transcript stabilization with ionomycin resulted in an increase of fluorescence that is quantitated by FACS analysis of responding cells. Using this system we have identified okadaic acid as a novel stabilizing compound, and investigated the upstream signaling pathways leading to stabilization. This reporter system has the advantage of speed and simplicity over standard methods currently in use and in addition to serving as a research tool it can be easily automated to increase throughput for drug discover

A novel homozygous SLC2A9 mutation associated with renal-induced hypouricemia

Hereditary renal hypouricemia (RHUC) is a genetically heterogenous disorder characterized by defective uric acid (UA) reabsorption resulting in hypouricemia and increased fractional excretion of UA; acute kidney injury (AKI) and nephrolithiasis are recognized complications. Type 1 (RHUC1) is caused by mutations in the SLC22A12 gene, whereas RHUC2 is caused by mutations in the SLC2A9 gene. Patient ethnicity is diverse but only few Caucasian families with an SLC2A9 mutation have been reported. The current report describes the clinical history, biochemical and molecular genetics findings of a native Austrian family with RHUC2. The propositus presented with 2 episodes of exercise-induced AKI and exhibited profound hypouricemia. Mutational screening of the SLC22A12 and SLC2A9 genes was performed. The molecular analyses revealed the homozygous c.512G>A transition that leads to the p.Arg171His missense substitution in SLC2A9, confirming the diagnosis of RHUC2. Segregation study of the causal mutation revealed that the mother and elder sister were heterozygous carriers, whereas the younger sister was found to be homozygous. We report the identification of a novel mutation in SLC2A9 as the cause of RHUC2 in a native Austrian family. We show that glucose transporter 9 mutations cause severe hypouricemia in homozygous individuals and confirm the high risk of AKI in male individuals harbouring these mutations. In our literature review, we provide an overview of the putative underlying pathophysiology, potential renal complications, findings on kidney biopsy as well as potential long-time renal sequela

mTORC2 sustains thermogenesis via Akt-induced glucose uptake and glycolysis in brown adipose tissue

Activation of non-shivering thermogenesis (NST) in brown adipose tissue (BAT) has been proposed as an anti-obesity treatment. Moreover, cold-induced glucose uptake could normalize blood glucose levels in insulin-resistant patients. It is therefore important to identify novel regulators of NST and cold-induced glucose uptake. Mammalian target of rapamycin complex 2 (mTORC2) mediates insulin-stimulated glucose uptake in metabolic tissues, but its role in NST is unknown. We show that mTORC2 is activated in brown adipocytes upon β-adrenergic stimulation. Furthermore, mice lacking mTORC2 specifically in adipose tissue (AdRiKO mice) are hypothermic, display increased sensitivity to cold, and show impaired cold-induced glucose uptake and glycolysis. Restoration of glucose uptake in BAT by overexpression of hexokinase II or activated Akt2 was sufficient to increase body temperature and improve cold tolerance in AdRiKO mice. Thus, mTORC2 in BAT mediates temperature homeostasis via regulation of cold-induced glucose uptake. Our findings demonstrate the importance of glucose metabolism in temperature regulation

Transcriptome-wide expression profiling in skin fibroblasts of patients with joint hypermobility syndrome/ehlers-danlos syndrome hypermobility type

Joint hypermobility syndrome/Ehlers-Danlos syndrome hypermobility type (JHS/EDS-HT), is likely the most common systemic heritable connective tissue disorder, and is mostly recognized by generalized joint hypermobility, joint instability complications, minor skin changes and a wide range of satellite features. JHS/EDS-HT is considered an autosomal dominant trait but is still without a defined molecular basis. The absence of (a) causative gene(s) for JHS/EDS-HT is likely attributable to marked genetic heterogeneity and/or interaction of multiple loci. In order to help in deciphering such a complex molecular background, we carried out a comprehensive immunofluorescence analysis and gene expression profiling in cultured skin fibroblasts from five women affected with JHS/EDS-HT. Protein study revealed disarray of several matrix structural components such as fibrillins, tenascins, elastin, collagens, fibronectin, and their integrin receptors. Transcriptome analysis indicated perturbation of different signaling cascades that are required for homeostatic regulation either during development or in adult tissues as well as altered expression of several genes involved in maintenance of extracellular matrix architecture and homeostasis (e.g., SPON2, TGM2, MMP16, GPC4, SULF1), cell-cell adhesion (e.g., CDH2, CHD10, PCDH9, CLDN11, FLG, DSP), immune/inflammatory/pain responses (e.g., CFD, AQP9, COLEC12, KCNQ5, PRLR), and essential for redox balance (e.g., ADH1C, AKR1C2, AKR1C3, MAOB, GSTM5). Our findings provide a picture of the gene expression profile and dysregulated pathways in JHS/EDS-HT skin fibroblasts that correlate well with the systemic phenotype of the patients

GLUT10 deficiency leads to oxidative stress and non-canonical αvβ3 integrin-mediated TGFβ signalling associated with extracellular matrix disarray in arterial tortuosity syndrome skin fibroblasts

Arterial tortuosity syndrome (ATS) is an autosomal recessive connective tissue disorder caused by loss-of-function mutations in SLC2A10, which encodes facilitative glucose transporter 10 (GLUT10). The role of GLUT10 in ATS pathogenesis remains an enigma, and the transported metabolite(s), i.e. glucose and/or dehydroascorbic acid, have not been clearly elucidated. To discern the molecular mechanisms underlying the ATS aetiology, we performed gene expression profiling and biochemical studies on skin fibroblasts. Transcriptome analyses revealed the dysregulation of several genes involved in TGFβ signalling and extracellular matrix (ECM) homeostasis as well as the perturbation of specific pathways that control both the cell energy balance and the oxidative stress response. Biochemical and functional studies showed a marked increase in ROS-induced lipid peroxidation sustained by altered PPARγ function, which contributes to the redox imbalance and the compensatory antioxidant activity of ALDH1A1. ATS fibroblasts also showed activation of a non-canonical TGFβ signalling due to TGFBRI disorganization, the upregulation of TGFBRII and connective tissue growth factor, and the activation of the αvβ3 integrin transduction pathway, which involves p125FAK, p60Src and p38 MAPK. Stable GLUT10 expression in patients' fibroblasts normalized redox homeostasis and PPARγ activity, rescued canonical TGFβ signalling and induced partial ECM re-organization. These data add new insights into the ATS dysregulated biological pathways and definition of the pathomechanisms involved in this disorder

Flexible logic locking obfuscation for sequential circuit through synchronous configurable blocks

LAUREA MAGISTRALECon la delocalizzazione del processo di produzione di dispositivi Hardware, i circuiti integrati sono stati esposti a molteplici attacchi. Il furto di proprietà intellettuale condotto dalla fabbrica che si occupa di produrli o la pratica di reverse engineering (RE) perpetrata da una design house sono solo alcuni esempi. In questo scenario, la ricerca accademica ha prodotto innumerevole materiale su possibili meccanismi di difesa e procedure d'attacco per sfruttarne le vulnerabilità. Negli ultimi anni, il logic locking, un insieme di tecniche di protezione basate sulla presenza di elementi programmabili, si è dimostrato versatile e applicabile a diversi livelli nel processo di sintesi. Il nostro lavoro può essere considerato come un passo verso una soluzione più dinamica, capace di combinare metodi difensivi per resistere sotto condizioni e vincoli dati. Il seguente testo propone un processo di integrazione tra due tecniche di logic locking già esistenti: Janus HD e latch-based obfuscation. In particolare, mira ad espandere il numero delle possibili chiavi del primo metodo introducendo blocchi configurabili basati sul latch. Successivamente, considerando la generazione di chiavi equivalenti che danno luogo alla stessa configurazione difensiva come il principale fattore di debolezza per la procedura di integrazione, si passa a valutare lo spazio delle chiavi della soluzione proposta e il suo variare rispetto ai principali parametri. Infine siamo in grado di specificare delle condizioni in cui la molteplicità introdotta non rappresenta una vulnerabilità per il circuito.Nowadays, the global spreading of the supply chain in Hardware production makes the integrated circuit (IC) design a target of multiple threats. The intellectual property (IP) piracy conducted by an untrusted foundry or the reverse engineering procedure (RE) brought by a malicious design house are only some examples. In this situation, many academic papers have produced different defense techniques and new attacks to exploit their vulnerabilities. In recent years, logic locking, a bunch of protection mechanisms based on inserting programmable elements inside the circuit, has shone for its versatility and applicability at different levels of abstraction. Our work could be considered a first step toward a more general solution that dynamically combines a subset of defense methods to obtain robust obfuscated circuitry under defined attack and constraint specifications. In our thesis, we propose an approach that integrates two existing logic locking techniques: the Janus HD and the latch-based obfuscation. In particular, we extend the key space of the first mechanism, inserting configurable memory elements based on the latch. Then, identifying the presence of equivalence keys as a critical vulnerability for our solution, we analyze their impact and specify the parameters that affect them. In the end, we define a range where the multiplicity introduced by the integration does not provide a weakness

The Effects Induced by Spinal Manipulative Therapy on the Immune and Endocrine Systems

Background andObjectives: Spinal manipulations are interventions widely used by different healthcare professionals for the management of musculoskeletal (MSK) disorders. While previous theoretical principles focused predominantly on biomechanical accounts, recent models propose that the observed pain modulatory effects of this form of manual therapy may be the result of more complex mechanisms. It has been suggested that other phenomena like neurophysiological responses and the activation of the immune-endocrine system may explain variability in pain inhibition after the administration of spinal manipulative therapy (SMT). The aim of this paper is to provide an overview of the available evidence supporting the biological plausibility of high-velocity, low-amplitude thrust (HVLAT) on the immune-endocrine system. Materials and Methods: Narrative critical review. An electronic search on MEDLINE, ProQUEST, and Google Scholar followed by a hand and "snowballing" search were conducted to find relevant articles. Studies were included if they evaluated the effects of HVLAT on participants' biomarkers Results: The electronic search retrieved 13 relevant articles and two themes of discussion were developed. Nine studies investigated the effects of SMT on cortisol levels and five of them were conducted on symptomatic populations. Four studies examined the effects of SMT on the immune system and all of them were conducted on healthy individuals. Conclusions: Although spinal manipulations seem to trigger the activation of the neuroimmunoendocrine system, the evidence supporting a biological account for the application of HVLAT in clinical practice is mixed and conflicting. Further research on subjects with spinal MSK conditions with larger sample sizes are needed to obtain more insights about the biological effects of spinal manipulative therapy

Small fiber neuropathy is a common feature of Ehlers-Danlos syndromes

To investigate the involvement of small nerve fibers in Ehlers-Danlos syndrome (EDS). Patients diagnosed with EDS underwent clinical, neurophysiologic, and skin biopsy assessment. We recorded sensory symptoms and signs and evaluated presence and severity of neuropathic pain according to the Douleur Neuropathique 4 (DN4) and ID Pain questionnaires and the Numeric Rating Scale (NRS). Sensory action potential amplitude and conduction velocity of sural nerve was recorded. Skin biopsy was performed at distal leg and intraepidermal nerve fiber density (IENFD) obtained and referred to published sex- and age-adjusted normative reference values. Our cohort included 20 adults with joint hypermobility syndrome/hypermobility EDS, 3 patients with vascular EDS, and 1 patient with classic EDS. All except one patient had neuropathic pain according to DN4 and ID Pain questionnaires and reported 7 or more symptoms at the Small Fiber Neuropathy Symptoms Inventory Questionnaire. Pain intensity was moderate (NRS ≥4 and <7) in 8 patients and severe (NRS ≥7) in 11 patients. Sural nerve conduction study was normal in all patients. All patients showed a decrease of IENFD consistent with the diagnosis of small fiber neuropathy (SFN), regardless of the EDS type. SFN is a common feature in adults with EDS. Skin biopsy could be considered an additional diagnostic tool to investigate pain manifestations in EDS

Constraining Warm Dark Matter using QSO gravitational lensing

Warm Dark Matter (WDM) has been invoked to resolve apparent conflicts of Cold Dark Matter (CDM) models with observations on subgalactic scales. In this work we provide a new and independent lower limit for the WDM particle mass (e.g. sterile neutrino) through the analysis of image fluxes in gravitationally lensed QSOs. Starting from a theoretical unperturbed cusp configuration we analyze the effects of intergalactic haloes in modifying the fluxes of QSO multiple images, giving rise to the so-called anomalous flux ratio. We found that the global effect of such haloes strongly depends on their mass/abundance ratio and it is maximized for haloes in the mass range 10^6-10^8 \Msun. This result opens up a new possibility to constrain CDM predictions on small scales and test different warm candidates, since free streaming of warm dark matter particles can considerably dampen the matter power spectrum in this mass range. As a consequence, while a ($\Lambda$)CDM model is able to produce flux anomalies at a level similar to those observed, a WDM model, with an insufficiently massive particle, fails to reproduce the observational evidences. Our analysis suggests a lower limit of a few keV ($m_{\nu} \sim 10$) for the mass of warm dark matter candidates in the form of a sterile neutrino. This result makes sterile neutrino Warm Dark Matter less attractive as an alternative to Cold Dark Matter, in good agreement with previous findings from Lyman-$\alpha$ forest and Cosmic Microwave Background analysis.Comment: One equation added, typo in eq: 5 corrected, minor changes to match the accepted version by MNRA