Purpose The study aims to investigate the interplay among digital technologies, intellectual capital and innovation. Thus far, there have been scant research on such intricate bundle of interactions. Also, the findings of previous studies were rather inconclusive, because conflicting results emerged over time. Building on the existence of heterogeneous evidences, this study solved the detected criticism by suggesting a curvilinear relationship among digital technologies, digital skills of human capital and intellectual property. Specifically, we argue that the relationship between digital technologies and intellectual property is inverted u-shaped. Design/methodology/approach Hypotheses are tested by applying a generalized linear model (GLM) regression analysis and a quadratic model for non-linear regression. The study analysed a large-scale sample of micro-data drawn from Eurostat. Such sample embraces the population of firms operating in all European member states. Findings Overall, the results of the study confirm that digital technologies are curvilinearly related to intellectual property. Precisely, the curve is inverted u-shaped. Notably, results show that digital skills only matter when employees have very demanding duties to accomplish. In all other cases, digital skills do not affect intellectual property significantly. Research limitations/implications The research is solely focused on firms' operating in the European Union. Future studies should extend the analysis to other geographies. Practical implications At a real impact level, the study suggests that intellectual property is only partially fostered by digital skills and digital technologies. In this sense, digital skills might be overrated. Originality/value Differently from prior research, this study originally detangles the impact of digital technologies on firm's intellectual capital by suggesting the existence of an inverse u-shaped relationship between variables.

The literature has neglected to analyse employer associations as organizations facing potential environmental threats to their financial sustainability. We examine associations' responses to collective bargaining decentralization, a major, contemporary threat. Using a qualitative, comparative case approach, we examine eight associations four each in Australia and Italy to develop a model of response types. Stronger decentralization effects increase associations' exposure to new and heightened competition, which in turn produces stronger association responses. These include prioritizing commercial over associational objectives. We analyse responses using strategic choice and resource dependence theories, finding that associations use both. However, the decision how to combine them reflects environmental conditions as well as choices linking organizational purpose and financial sustainability.

This work describes the effectiveness of HDAC-inhibitor (S)-2 towards colorectal cancer (CRC) HCT116 cells in vitro by inducing cell cycle arrest and apoptosis, and in vivo by contrasting tumour growth in mice xenografts. Among the multifaceted drug-induced events described herein, an interesting link has emerged between the oncoprotein histone deacetylase HDAC1 and the oncogenic Cancerous Inhibitor of Protein Phosphatase 2A (CIP2A) which is overexpressed in several cancers including CRCs. HDAC1 inhibition by (S)-2 or specific siRNAs downregulates CIP2A transcription in three different CRC cell lines, thus restoring the oncosuppressor phosphatase PP2A activity that is reduced in most cancers. Once re-activated, PP2A dephosphorylates pGSK-3β(ser9) which phosphorylates β-catenin that remains within the cytosol where it undergoes degradation. The decreased amount/activity of β-catenin transcription factor prompts cell growth arrest by diminishing c-Myc and cyclin D1 expression and abrogating the prosurvival Wnt/β-catenin signaling pathway. These results are the first evidence that the inhibition of HDAC1 by (S)-2 downregulates CIP2A transcription and unleashes PP2A activity, thus inducing growth arrest and apoptosis in CRC cells.

Histone deacetylase inhibitors ( HDAC i) are agents capable of inducing growth arrest and apoptosis in different tumour cell types. Previously, we reported a series of novel HDAC i obtained by hybridizing SAHA or oxamflatin with 1,4‐benzodiazepines. Some of these hybrids proved effective against haematological and solid cancer cells and, above all, compound (S)‐8 has emerged for its activities in various biological systems. Here, we describe the effectiveness of (S)‐8 against highly metastatic human A375 melanoma cells by using normal PIG 1 melanocytes as control. (S)‐8 prompted: acetylation of histones H3/H4 and α‐tubulin; G 0 /G 1 and G 2 /M cell cycle arrest by rising p21 and hypophos‐phorylated RB levels; apoptosis involving the cleavage of PARP and caspase 9, BAD protein augmentation and cytochrome c release; decrease in cell motility, invasiveness and pro‐angiogenic potential as shown by results of wound‐healing assay, down‐regulation of MMP ‐2 and VEGF ‐A/ VEGF ‐R2, besides TIMP ‐1/ TIMP ‐2 up‐regulation; and also intracellular accumulation of melanin and neutral lipids. The pan‐caspase inhibitor Z‐ VAD ‐fmk, but not the antioxidant N‐acetyl‐cysteine, contrasted these events. Mechanistically, (S)‐8 allows the disruption of cytoplasmic HDAC 6‐protein phosphatase 1 ( PP 1) complex in A375 cells thus releasing the active PP 1 that dephosphorylates AKT and blocks its downstream pro‐survival signalling. This view is consistent with results obtained by: inhibiting PP 1 with Calyculin A; using PPP 1R2‐transfected cells with impaired PP 1 activity; monitoring drug‐induced HDAC 6‐ PP 1 complex re‐shuffling; and, abrogating HDAC 6 expression with specific si RNA . Altogether, (S)‐8 proved very effective against melanoma A375 cells, but not normal melanocytes, and safe to normal mice thus offering attractive clinical prospects for treating this aggressive malignancy.

There is great interest in developing reliable biomarkers to support antemortem diagnosis of late-onset Alzheimer's disease (AD). Early prediction and diagnosis of AD might be improved by the detection of a proteolytic dysfunction in extracts from cultured AD fibroblasts, producing altered isoelectrophoretic forms of the enzyme transketolase (TK-alkaline bands). The TK profile and apolipoprotein E (APOE) genotype were examined in fibroblasts from 36 clinically diagnosed probable late-onset sporadic AD patients and 38 of their asymptomatic relatives, 29 elderly healthy individuals, 12 neurological non-AD patients, and 5 early-onset AD patients. TK alterations occurred in (i) several probable AD patients regardless of age-of-onset and severity of disease; (ii) all early-onset AD patients and APOE ε 4/4 carriers; and (iii) nearly half of asymptomatic AD relatives. Normal subjects and non-AD patients were all negative. Notably, culture conditions promoting TK alterations were also effective in increasing active BACE1 levels. Overall, the TK assay might represent a low-cost laboratory tool useful for supporting AD differential diagnosis and identifying asymptomatic subjects who are at greater risk of AD and who should enter a follow-up study. Moreover, the cultured fibroblasts were confirmed as a useful in vitro model for further studies on the pathogenetic process of AD.

A series of new histone deacetylase inhibitors were designed and synthesized based on hybridization between SAHA or oxamflatin and 5-phenyl-1,4-benzodiazepines. The compounds were tested for their enzyme inhibitory activity on HeLa nuclear extracts, and on human recombinant HDAC1 and HDAC6. Antiproliferative activity was tested on different cancer cells types, while proapoptotic activity was primarily tested on NB4 cells. The compounds showed IC50 values similar to those of SAHA. Compound (S)-8 displayed interesting activity against hematological and solid malignancies.

Histone deacetylase inhibitors (HDACi) represent a promising class of epigenetic agents with anticancer properties. Here, we report that (S)-2, a novel hydroxamate-based HDACi, shown previously to be effective against acute myeloid leukemia cells, was also a potent inducer of apoptosis/differentiation in human prostate LNCaP and PC3 cancer cells. In LNCaP cells (S)-2 was capable of triggering H3/H4 histone acetylation, H2AX phosphorylation as a marker of DNA damage and producing G0/G1 cell cycle arrest. Consistently, (S)-2 led to enhanced expression of both the protein and mRNA p21 levels in LNCaP cells but, contrary to SAHA, not in normal non-tumorigenic prostate PNT1A cells. Mechanistic studies demonstrated that (S)-2-induced apoptosis in LNCaP cells developed through the cleavage of pro-caspase 9 and 3 and of poly(ADP-ribose)-polymerase accompanied by the dose-dependent loss of mitochondrial membrane potential. Indeed, the addition of the pan-caspase inhibitor Z-VAD-fmk greatly reduced drug-mediated apoptosis while the antioxidant N-acetyl-cysteine was virtually ineffective. Importantly, preliminary data with nude mice xenografted with LNCaP cells showed that (S)-2 prompted a decrease in the tumor volume and an increase in H2AX phosphorylation within the cancer cells. Moreover, the highly metastatic prostate cancer PC3 cells were also sensitive to (S)-2 that: i) induced growth arrest and moderate apoptosis; ii) steered cells towards differentiation and neutral lipid accumulation; iii) reduced cell invasiveness potential by decreasing the amount of MMP-9 activity and up-regulating TIMP-1 expression; and iv) inhibited cell motility and migration through the Matrigel. Overall, (S)-2 has proven to be a powerful HDACi capable of inducing growth arrest, cell death and/or differentiation of LNCaP and PC3 prostate cancer cells and, due to its low toxicity and efficacy in vivo, might also be of clinical interest to support conventional prostate cancer therapy.

In this paper we investigate the development over time of employers’ associations as a peculiar form of meta-organization. In particular, we try to understand how in the last decades the progressive decentralization of the collective bargaining and the reshaping of the economic landscape impacted on employers’ associations by increasing their members’ heterogeneity, and how this has affected their trajectories of evolution, internal functioning and identity. We qualitatively study the case of the largest territorial employers’ organization in Italy (Assolombarda): in the last decades new members were recruited almost exclusively among small size firms and in the tertiary sector; this resulted in an increased heterogeneity of the members’ composition which has lead to different expectations and a new definition of the association mission. The provision of ‘elective’ services as become a major source of financial resources, and a growing number of members now evaluate the overall offer of inducements (either as elective or as selective services) as a key factor in their decision to join and remain. The organizational identity is a key element of many ‘partial organization’ and it is protected by deciding who is allowed to join: by weakening this element, Assolombarda is losing its capability to organize the collective action of the members and it is in fact evolving – even if key actors do not appear fully conscious of that consciously – by increasingly reassembling to a form of business network.

Histone deacetylase inhibitors (HDACi) induce tumour cell cycle arrest and/or apoptosis, and some of them are currently used in cancer therapy. Recently, we described a series of powerful HDACi characterized by a 1,4‐benzodiazepine (BDZ) ring hybridized with a linear alkyl chain bearing a hydroxamate function as Zn++‐chelating group. Here, we explored the anti‐leukaemic properties of three novel hybrids, namely the chiral compounds (S)‐2 and (R)‐2, and their non‐chiral analogue 4, which were first comparatively tested in promyelocytic NB4 cells. (S)‐2 and partially 4– but not (R)‐2 – caused G0/G1 cell‐cycle arrest by up‐regulating cyclin G2 and p21 expression and down‐regulating cyclin D2 expression, and also apoptosis as assessed by cell morphology and cytofluorimetric assay, histone H2AX phosphorylation and PARP cleavage. Notably, these events were partly prevented by an anti‐oxidant. Moreover, novel HDACi prompted p53 and α‐tubulin acetylation and, consistently, inhibited HDAC1 and 6 activity. The rank order of potency was (S)‐2 > 4 > (R)‐2, reflecting that of other biological assays and addressing (S)‐2 as the most effective compound capable of triggering apoptosis in various acute myeloid leukaemia (AML) cell lines and blasts from patients with different AML subtypes. Importantly, (S)‐2 was safe in mice (up to 150 mg/kg/week) as determined by liver, spleen, kidney and bone marrow histopathology; and displayed negligible affinity for peripheral/central BDZ‐receptors. Overall, the BDZ‐hydroxamate (S)‐2 showed to be a low‐toxic HDACi with powerful anti‐proliferative and pro‐apototic activities towards different cultured and primary AML cells, and therefore of clinical interest to support conventional anti‐leukaemic therapy.

Cholesterol is an essential constituent of all mammalian cell membranes and its availability is therefore a prerequisite for cellular growth and other functions. Several lines of evidence are now indicating an association between alterations of cholesterol homeostasis and cell cycle progression. However, the role of cholesterol in cell differentiation is still largely unknown. To begin to address this issue, in this study we examined changes in cholesterol metabolism and in the mRNA levels of proteins involved in cholesterol import and esterification (multi-drug resistance, MDR-3) and acylCoA: cholesterol acyltransferase (ACAT) and cholesterol export (caveolin-1) in Friend virus-induced erythroleukemia cells (MELC), in the absence or in the presence of the chemical inducer of differentiation, hexamethylene bisacetamide (HMBA). FBS-stimulated growth of MELC was accompanied by an immediate elevation of cholesterol synthesis and cholesterol esterification, and by an increase in the levels of MDR-3 and ACAT mRNAs. A decrease in caveolin-1 expression was also observed. However, when MELC were treated with HMBA, the inhibition of DNA synthesis caused by HMBA treatment, was associated with a decrease in cholesterol esterification and in ACAT and MDR-3 mRNA levels and an increase in caveolin-1 mRNA. Detection of cytoplasmic neutral lipids by staining MELC with oil red O, a dye able to evidence CE but not FC, revealed that HMBA-treatment also reduced growth-stimulated accumulation of cholesterol ester to approximately the same extent as the ACAT inhibitor, SaH. Overall, these results indicate for the first time a role of cholesterol esterification and of some related genes in differentiation of erythroid cells.

Cholesterol is an essential constituent of all mammalian cell membranes, and its availability is therefore a prerequisite for cellular growth and other functions. Several lines of evidence are now indicating an association between alterations of cholesterol homeostasis and cell cycle progression in cancer cells. However, the role of cholesterol in cell differentiation is still largely unknown. To begin to address this issue, in this study we examined changes in cholesterol metabolism and in the mRNA levels of proteins involved in cholesterol import and esterification (multi-drug resistance, MDR-3) and acylCoA:cholesterol acyltransferase (ACAT) and cholesterol export (caveolin-1) in Friend virus-induced erythroleukemia cells (MELC), in the absence or in the presence of the chemical inducer of differentiation, hexamethylene bisacetamide (HMBA). In uninduced MELC, cholesterol synthesis, esterification and MDR-3 and ACAT mRNAs increased as cells progressed from resting to proliferating phase, while caveolin-1 decreased. These results provide evidence that cholesterol esterification “per se” may have a role in cell division. When MELC are treated with HMBA, the reduction of DNA synthesis caused by the inducer is accompanied by an extensive decrease of cholesterol esterification and of ACAT and MDR-3 mRNA levels and by a significant increase in caveolin-1 mRNA. On the other hand, detection of cytoplasmic neutral lipids by staining MELC with oil-ORO, a dye able to evidence CE but not FC, revealed that HMBA-treatment inhibits cholesterol ester accumulation in MELC to approximately the same extent as the ACAT inhibitor, SaH. These results, other than, to add new insights on the possible role of cholesterol metabolism during tumor growth, for the first time indicate a possible involvement of cholesterol esterification pathways in the regulating of differentiation of erythroleukemic cells.

Objective. This study aimed to investigate the mechanisms of action of WEB-2170, an inverse agonist of platelet-activating factor receptor, capable of inducing apoptosis in human acute myelogenous leukemia (AML) cells.Material and Methods. Gene expression profiling followed by cytofluorimetric, morphologic, and biologic analyses were used to monitor WEB-2170 effects in AML cell lines (ie, NB4, KG1, NB4-MR4, THP1, and U937) and blasts from patients with different ANIL (M0-M5) subtypes. PTEN silencing with small interfering RNA was also performed.Results. We have demonstrated that drug-mediated cytostasis/apoptosis in NB4 cells is characterized by upregulation of cyclin G2, p21/WAF1, NIX, TNF-alpha, and PTEN expression, and downregulation of cyclin D2 and BCL2 expression. We observed an increase in PTEN protein accompanied by a decrease in phospho-extracellular signal-regulated kinase 2 (ERK2) and phospho-AKT, and by forkhead box O3a (FOXO3a) cytoplasmic-nuclear translocation; the mitochondrial cytochrome C release and PARP cleavage marked the late apoptotic steps. We have found that WEB-2170 triggered apoptosis in NB4, KG1, and NB4-MR4 cells where PTEN was expressed, but not in THP1 and U937 cells where PTEN was absent. Finally, we show that PTEN silencing in NB4 cells by PTEN-specific small interfering RNA resulted in a significant reduction of drug-induced apoptosis.Conclusion. We demonstrated that WEB-2170 is a powerful antileukemic agent with interesting translational opportunities to treat AML and described mechanisms of drug-induced intrinsic and extrinsic apoptosis both in AML cell lines and blasts from AML patients by addressing PTEN as the master regulator of the whole process. (C) 2009 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc.

Decentralisation of collective bargaining generates challenges to employer associations’ organisational purpose, identity and strategies for survival and growth. We focus on one important strategic choice through which associations may seek to address challenges of membership and financial sustainability: whether to substitute (even partially) traditional purposes and identities as representative associations by embracing those of commercial businesses in the broader business services market. Our study develops a cross-national, comparative analysis of eight associations, four in Australia and four in Italy. We find that labour market and product market pressures most important for explaining these choices. Acknowledgments. We warmly thank all the employer association officials who have helped us over many years and, in particular, Dr Laura Mengoni (Assolombarda) for her help on Italy .

Abstract Transcriptional silencing via promoter methylation of genes important for cell growth and differentiation plays a key role in myeloid leukemogenesis. We find that clinically achievable levels of 5-aza-2′-deoxycytidine (5-AZA-dC), a potent inhibitor of DNA methylation, can modify chromatin and restore the ability of tumor necrosis factor α (TNFα) to induce monocytic differentiation of the acute myeloid leukemia cells NB4 and U937. Although 5-AZA-dC cannot fully induce differentiation, we show that 5-AZA-dC acts directly on TNFα-responsive promoters to facilitate TNFα-induced transcriptional pathways leading to differentiation. 5-AZA-dC regulates the expression of Dif-2, a TNFα target gene, by deacetylating chromatin domains in a methylation-dependent manner. Chromatin immunoprecipitation analyses of the Dif-2 promoter show histone hyperacetylation and a recruitment of the nuclear factor-κB transcription factor in response to 5-AZA-dC. Furthermore, 5-AZA-dC plus TNFα enhances the level of phosphorylated RNA Pol II at the Dif-2 promoter via synergistic recruitment of TFIIH. We conclude that nonspecific changes in chromatin can allow a specific transcriptional inducer to overcome blocks in leukemic cell differentiation. Our results support the concept of low doses of 5-AZA-dC acting in combination with other agents to target epigenetic changes that drive malignant growth in leukemic cells. [Cancer Res 2009;69(1):55–64]

Intracellular cholesterol metabolism was reported to modulate amyloid-beta (Abeta) generation in Alzheimer's disease (AD). Results presented herein demonstrated that, like brain cells, cultured skin fibroblasts from AD patients contained more cholesterol esters than fibroblasts from healthy subjects. Particularly, Oil Red-O, Nile Red, and filipin staining highlighted higher levels of neutral lipids which responded to inhibitors of acyl-coenzyme A:cholesterol acyl-transferase (ACAT-1), associated with an increase in free cholesterol. ACAT-1 mRNA levels increased significantly in AD fibroblasts, whereas those of sterol regulatory element binding protein-2, neutral cholesterol ester hydrolase, and ATP-binding cassette transporter member 1 were markedly down-regulated. Instead, mRNA levels of low-density lipoprotein receptor, hydroxy-methyl-glutaryl-coenzyme A reductase, caveolin-1, and amyloid-beta protein precursor (AbetaPP) were virtually unchanged. Notably, mRNA levels of both beta-site AbetaPP-cleaving enzyme 1 (BACE1) and neprilysin were significantly down-regulated. An increase in Abeta(40) and Abeta(42) immunostaining and a decrease in BACE1 active form were also found in AD versus control fibroblasts. Altogether, these findings support the hypothesis that the derangement of cholesterol homeostasis is a systemic alteration involving central but also peripheral cells of AD patients, and point to cholesterol ester levels in AD fibroblasts as an additional metabolic hallmark useful in the laboratory and clinical practice.

Superior vena cava (SVC) occlusion can be clinically recognized in the acute setting when the stenosing process does not allow the development of collateral venous channels, which guarantee the venous drainage to the right heart. On the contrary, when the obstruction develops progressively, the diagnosis of SVC obstruction may remain undiagnosed. In the present case, the presence of SVC thrombosis was purely coincidental. In fact, the obstruction was first noticed on diagnostic tests performed because of the malfunction of a totally implantable Porth a Cath placed into the superior vena cava (through right subclavian access), five years before, in a patient suffering from non-Hodgkin disease. Venography is the most appropriate diagnostic methodology which reveals the presence of a dilated azygos vein as a compensatory mechanism. Comparison with computed tomography allows to confirm the diagnosis and to identify the possible causes. Dilatation of the azygos vein, secondary to superior vena cava thrombosis, although a rare event, should be taken into consideration in those patients with CVC and who present with frequent episodes of deep venous thrombosis.

Background: Alzheimer's disease is the most common progressive neurodegenerative disease. In recent years, numerous progresses in the discovery of novel Alzheimer's disease molecular biomarkers in brain as well as in biological fluids have been made. Among them, those involving lipid metabolism are emerging as potential candidates. In particular, an accumulation of neutral lipids was recently found by us in skin fibroblasts from Alzheimer's disease patients. Therefore, with the aim to assess whether peripheral alterations in cholesterol homeostasis might be relevant in Alzheimer's disease development and progression, in the present study we analyzed lipid metabolism in plasma and peripheral blood mononuclear cells from Alzheimer's disease patients and from their first-degree relatives.Methods: Blood samples were obtained from 93 patients with probable Alzheimer's disease and from 91 of their first-degree relatives. As controls we utilized 57, cognitively normal, over-65 year-old volunteers and 113 blood donors aged 21-66 years, respectively. Data are reported as mean +/- standard error. Statistical calculations were performed using the statistical analysis software Origin 8.0 version. Data analysis was done using the Student t-test and the Pearson test.Results: Data reported here show high neutral lipid levels and increased ACAT-1 protein in about 85% of peripheral blood mononuclear cells freshly isolated (ex vivo) from patients with probable sporadic Alzheimer's disease compared to about 7% of cognitively normal age-matched controls. A significant reduction in high density lipoprotein-cholesterol levels in plasma from Alzheimer's disease blood samples was also observed. Additionally, correlation analyses reveal a negative correlation between high density lipoprotein-cholesterol and cognitive capacity, as determined by Mini Mental State Examination, as well as between high density lipoprotein-cholesterol and neutral lipid accumulation. We observed great variability in the neutral lipid-peripheral blood mononuclear cells data and in plasma lipid analysis of the subjects enrolled as Alzheimer's disease-first-degree relatives. However, about 30% of them tend to display a peripheral metabolic cholesterol pattern similar to that exhibited by Alzheimer's disease patients.Conclusion: We suggest that neutral lipid-peripheral blood mononuclear cells and plasma high density lipoprotein-cholesterol determinations might be of interest to outline a distinctive metabolic profile applying to both Alzheimer's disease patients and asymptomatic subjects at higher risk of disease.