bluuebunny/crossref_metadata_2025 huggingface dataset:
The travelling business of sustainable urbanism: International consultants as norm-setters
Authors: Rapoport Elizabeth , Hult Anna
Published: January 2017

This article examines the international travels of ideas about sustainable urban planning and design through a focus on private sector architecture, planning and engineering consultants. These consultants, who we refer to as the global intelligence corps (GIC), package up their expertise in urban sustainability as a marketable commodity, and apply it on projects around the world. In doing so, the global intelligence corps shape norms about what constitutes ‘good’ sustainable urban planning, and contribute to the development of an internationalised travelling model of sustainable urbanism. This article draws on a broad study of the industry (GIC) in sustainable urban planning and design, and two in-depth case studies of Swedish global intelligence corps firms working on Chinese Eco-city projects. Analysis of this material illustrates how the global intelligence corps’s work shapes a traveling model of sustainable urbanism, and how this in turn creates and reinforces particular norms in urban planning practice.
The practice of scalecraft: Scale, policy and the politics of the market in England’s academy schools
Authors: Papanastasiou Natalie
Published: January 2017

This paper builds on geographical understandings of scalar practices and illustrates how they can enrich studies of policymaking, in particular for the area of education policy. It achieves this by integrating a focus on ‘scalecraft’ with an approach to ‘policy as practice’ featuring in critical policy studies. The paper draws on one of the few empirical studies of England’s academy schools policy and analyses the work of actors tasked with implementing the policy in two local authority case studies. Analysis presents a new critical perspective on the academies policy by revealing how policy actors’ work is underpinned by scalecraft practices. The analysis also reveals how scalecraft intersects with marketisation politics to create scalar tensions which profoundly shape the work of policy actors. The paper uses its analytical findings to conceptually develop the relationship between scalar practices and the politics of the market, and to propose new conceptual dimensions for understanding the practice of scalecraft.
A review of the spatial distribution of and analytical techniques used in paleotempestological studies in the western North Atlantic Basin
Authors: Oliva Frank , Peros Matthew , Viau André
Published: January 2017

Paleotempestology, the study of past tropical cyclones (TCs) using geological proxy techniques, is a growing discipline that utilizes data from a broad range of sources. Most paleotempestological studies have been conducted using “established proxies,” such as grain-size analysis, loss-on-ignition, and micropaleontological indicators. More recently, however, researchers have been applying more advanced geochemical analyses, such as X-ray fluorescence core scanning and stable isotopic geochemistry, to generate new paleotempestological records. In this paper we begin by providing a list of paleotempestological studies for the western North Atlantic Basin and illustrate the spatial coverage of these studies. We then review the premises behind both established and new proxies and discuss their strengths and limitations at resolving past hurricane activity. Lastly, we suggest future directions for paleotempestological research based on our review of the literature that we argue will ultimately lead to a better understanding of TC dynamics under future climate change scenarios.
Differential Effects of Hydroxamate Inhibitors on PMA- and Ligand-Induced L-Selectin Down-Modulation: Role of Membrane Proximal and Cytoplasmic Domains
Authors: Jasuja R. R. , Mier J. W.
Published: January 2017

L-selectin is down-modulated from the cell surface in response to leukocyte activation or cross-linking with ligand mimetics such as anti-L-selectin antibodies or sulfatides. The down-modulation induced upon cellular activation with PMA is due to proteolytic shedding mediated by a cysteine metalloprotease, presumably the TNF-α converting enzyme (TACE), and is susceptible to inhibitors of the hydroxamate class. To determine if cross-linking-induced down-modulation of L-selectin is similarly affected by these inhibitors, we exposed Jurkat T cells to the anti-L-selectin antibody Dreg 200 or sulfatides in the presence of the hydroxamate TNF-α protease inhibitor (TAPI). TAPI completely inhibited PMA-induced down-modulation but had no effect on that induced by the anti-L-selectin antibody or sulfatides. The down-modulation induced by an anti-TCR antibody (WT31) was only partially inhibited by TAPI. An L-selectin mutant lacking the putative TACE cleavage site and resistant to PMA-induced shedding (321del.9) was expressed in L1.2 pre-B cells. Like WT L-selectin, this mutant was down-modulated when exposed to sulfatides, or Dreg 200 and this down-modulation was unaffected by TAPI. An L-selectin cytoplasmic tail deletion mutant (344del.15) expressed in L1.2 pre-B cells was down-modulated by PMA or sulfatides, but not Dreg 200. Electrophoretic analysis of L-selectin immunoprecipitated from the supernatants of Jurkat cells treated with either sulfatides or D200 revealed a proteolytic fragment of the same size as that released from the cell surface in response to PMA. Our data indicate that the down regulation of L-selectin in response to cross-linking by ligands or TCR engagement may be mechanistically distinct from that induced by PMA. Furthermore, our results with the 344del.15 mutant suggest that the down-modulation of L-selectin induced by certain sulfated carbohydrate ligands may be initiated through surface receptors other than L-selectin itself.
Carbamazepine-Induced Hypersensitivity Syndrome in a Child with Epilepsy
Authors: Verrotti A. , Feliciani C. , Morresi S. , Coscione G. , Morgese G. , Toto P. , Chiarelli F.
Published: January 2017

Carbamazepine is an effective anticonvulsant and is considered the drug of first choice for the treatment of partial and secondarily generalized seizures. Although carbamazepine is well tolerated, many side effects have been reported in the literature. The majority of these adverse effects are transient and do not lead to the discontinuation of the therapy.

We present a case of a female child, aged 11 years and 6 months, who showed an anticonvulsant hypersensitivity syndrome induced by carbamazepine. This syndrome is a rare, potentially life-threatening adverse drug reaction. The patient developed a cutaneous nonpruritic rash, associated with high fever, diffuse lymphadenopathy, and arthralgias on the knees and the ankles with local signs of arthritis. Laboratory examination showed a lymphocytosis, mild thrombocytopenia, marked eosinophilia, and high transaminases.

Corticosteroid therapy (betametasone 0,5 mg × 3 day) was started and carbamazepine was gradually withdrawn changing to valproic acid, with complete control of the seizures. The fever and the rash reduced gradually, beginning from the face and then disappearing completely after 10 days. Laboratory results showed a clear improvement: after 7 days the patient showed a complete normalization of the above parameters, except for transaminases. The complete normalization of these enzymes was observed after 2 weeks from the disappearance of the skin rash.
Morning Sudden Cardiac Death
Authors: Guagnano M. T. , Davì G. , Sensi S.
Published: January 2017

A circadian variation of cardiac function with peak in the early morning was documented about twenty years ago. A circadian rhythm of platelet aggregability, in the same time of the day, was demonstrated in healthy young male subjects. The morning hours were also reported as crucial for sympathetic nervous system activity, for heart rate variability, and for the abrupt rise in blood pressure. Altogether, these trigger factors may explain the high incidence of sudden cardiac death during the morning.

In the primary prevention of sudden death in patients with high cardiovascular risk, many strategies were proposed, such as implantable cardioverter-defibrillators, antiarrhythmic and antihypertensive therapies, particularly beta-blockers and more recently, aspirin. Also in subjects without cardiovascular risk factors, it is predictable that early and continuous administration of low-dose aspirin, by inhibiting platelet aggregation and thrombin formation, particularly in morning hours, may represent an effective therapy for the prevention of myocardial infarction and morning sudden cardiac death.
Role of Pegylated Lyposomal Doxorubicin (PLD) in Systemic Kaposi's Sarcoma: A Systematic Review
Authors: Di Trolio R. , Di Lorenzo G. , Delfino M. , De Placido S.
Published: January 2017

Kaposi's sarcoma (KS) is a form of skin cancer that can involve internal organs. It is often found in patients with acquired immunodeficiency syndrome (AIDS) and can be fatal. Kaposi's sarcoma produces pink, purple or brown tumors on the skin, mucous membranes or internal organs. Treatment goals for KS are simple: to reduce the severity of the symptoms, shrink tumors and prevent disease progression. Unfortunately, there is no single best treatment-plan that can achieve all these goals. With widespread KS lesions over the body surface or evidence of spreading to other parts of the body, the physicians need to treat the patients with systemic chemotherapy. A new class of drugs, called liposomal anthracyclines, appears to produce good results with fewer toxic side effects than more conventional cytotoxic drugs. One of these drugs, pegylated liposomal doxorubicin (PLD) has become the treatment of choice. This article summarizes all the studies with PLD in systemic Kaposi's sarcoma.
In Vitro Effects of Aminobisphosphonates on Vγ9Vδ2 T Cell Activation and Differentiation
Authors: Ferlazzo V. , Sferrazza C. , Caccamo N. , Di Fede G. , Di Lorenzo G. , D'Asaro M. , Meraviglia S. , Dieli F. , Rini G. , Salerno A.
Published: January 2017

In this study we have evaluated the in vitro effects of four different aminobisphosphonates, alendronate, risedronate, neridronate and zoledronate, on Vγ9Vδ2 T cell activation and differentiation. All tested aminobisphosphonates induce an IL-2-dependent activation and expansion of Vγ9Vδ2 T lymphocytes in primary PBMC cultures of healthy donors. Most notably, they also determine a different distribution of Vγ9Vδ2 T cell subsets, with decrease of Tnaive and TCM cells and increase of TEM and TEMRA Vγ9Vδ2 cells, indicating that in vitro treatment with aminobisphosphonates induces Vγ9Vδ2 T lymphocytes to differentiate towards an effector/cytotoxic phenotype. Accordingly, Vγ9Vδ2 T lymphocytes cultured with aminobisphosphonates and IL-2 showed a major content of IFN-γ and acquired the ability to kill tumor target cells.
Low Levels of Serum Cholesterol/Phospholipids are Associated with the Antiphospholipid Antibodies in Monoclonal Gammopathy
Authors: Pugliese L. , Bernardini I. , Viola-Magni M.P. , Albi E.
Published: January 2017

A decrease in cholesterol blood level, not due to a decrease synthesis by the liver, has been observed in patients suffering from tumors. In this work cholesterol blood was evaluated in patients affected by monoclonal gammopathy who were not subjected to any treatment. The blood of 25 patients were analyzed for protein and lipid content. Patients were divided according to the gamma protein content into three groups, and it was demonstrated that the group with high levels of gamma proteins presented a strong decrease in blood cholesterol and phospholipids. In these patients the presence of antibodies against phospholipids by using cardiolipin and phosphatidylinositol as antigens has also been demonstrated. The antibodies were rare in patients with a low content of gamma proteins and normal level of lipids, but the frequency was more than 80% in patients with low blood lipid levels.
Cox-Inhibitors Relieve the Immunosuppressive Effect of Tumor Cells and Improve Functions of Immune Effectors
Authors: Lang S. , Picu A. , Hofmann T. , Andratschke M. , Mack B. , Moosmann A. , Gires O. , Tiwari S. , Zeidler R.
Published: January 2017

A common phenomenon in cancer patients is a suppressed cell-mediated immunity, characterized by the inability of immune effector cells to mount efficient anti-tumor responses. Immunosuppressive factors, released by the tumor, contribute to this phenomenon and thus to tolerance. Prostaglandins, catalyzed by the cyclooxygenases (COX-1 and COX-2) from arachidonic acid, are one class of these factors. Since at least one of the COX enzymes is often expressed at high level in human cancers, the enzymes were ascribed a causal role in tumor etiology and progression. Non-steroidal antiinflammatory drugs (NSAIDs) like aspirin, which block COX activity, have demonstrated their antitumor effects in preclinical and clinical trials. Pro-apoptotic and anti-angiogenic effects in tumor cells may account for this activity. In addition, by inhibiting the release of prostaglandins from the tumor and by blocking COX activity in immune effector cells, NSAIDs may also bias the function of immune cells towards a more tumoricidal phenotype. We show here that tumor cells inhibit the physiological function of immune cells, and that NSAIDs restore this function. These data contribute to an understanding of the antineoplastic effect ascribed to NSAIDs and support the prophylactic use of these drugs in high-risk patients.
Bone Marrow Histopathological and Molecular Changes of Small B-Cell Lymphomas after Rituximab Therapy: Comparison with Clinical Response and Patients' Outcome
Authors: Goteri G. , Olivieri A. , Ranaldi R. , Lucesole M. , Filosa A. , Capretti R. , Pieramici T. , Leoni P. , Rubini C. , Fabris G. , Lo Muzio L.
Published: January 2017

This study correlates bone marrow changes after Rituximab (RTX) treatment with the clinical characteristics and outcome of 26 patients with small B-cell lymphomas. The percentage, phenotypic profile and clonality pattern of bone marrow lymphoid infiltrate were analysed before and after RTX treatment. Clinical, histological and molecular responses to RTX were correlated to the clinical outcome of the patients. Sixteen out of twenty-six patients obtained a complete clinical remission (CR). A favourable histology - follicular lymphoma (FL), hairy cell leukaemia (HCL) and marginal zone lymphoma (MZL) - was associated with a higher frequency of clinical CR and histological remission (HR), in comparison with mantle cell lymphoma (MCL), chronic lymphocytic leukaemia (CLL) and lymphoplasmacytic lymphoma (LPL). Two patterns of bone marrow HR were observed: 1) complete lymphoid cell disappearance (9 patients); or 2) nodular/interstitial T-cell infiltration (10 patients). Three histological persistence (HP) patterns were observed: 1) persistence of CD20+ small lymphoid cells in 1 patient with MCL; 2) loss of CD20 antigen expression in 4 patients with CLL; or 3) persistence only of clusters of monotypic plasma cells in 2 patients with LPL. CR and HR were strongly correlated. The percentage of lymphomatous infiltrate after RTX was higher in patients who subsequently died of the disease. Molecular response showed no correlations with the further clinical course in 12 patients achieving a complete clinical remission. In conclusion, bone marrow morphological and immunohistochemical analysis with a restricted panel of antibodies is useful to avoid 42% false positive and 85% false negative interpretations. Persistence of monoclonality after RTX might have a role in evaluating the molecular pattern of CD20-negative clones that can emerge after RTX as a tumoral escape to therapy.
Sjogren's Syndrome in a Celiac Patient: Searching for Environmental Triggers
Authors: D'Onofrio F. , Miele L. , Diaco M. , Santoro L. , De Socio G. , Montalto M. , Grieco A. , Gasbarrini G. , Manna R.
Published: January 2017

In the last few years many studies have shown the potential role of different triggers in the pathogenesis of several autoimmune diseases. In particular, in Sjogren's syndrome the presence of a genetic background is considered determining, but environmental factors have recently been described as triggers or precipitators. In this report, we describe the case of a young woman affected by celiac disease in which an Ascaris lumbricoides infestation and estrogen therapy could have played a role in the development of Sjogren's syndrome.
Interstitial Lung Disease in Systemic Sclerosis
Authors: Marasini B. , Belloli L. , Massarotti M.
Published: January 2017

Interstitial lung disease is an early and serious complication of systemic sclerosis (SSc). Because it may be asymptomatic for a long period, and only the early (logistic phase is at present susceptible to treatment, early diagnosis and identification of risk are critical to the outcome. However, identifying SSc patients at risk for developing interstitial lung disease is at present difficult; therefore, a strict monitoring of the disease, especially in the first years, is mandatory. Treatment strategy is aimed at suppressing inflammation. Unfortunately, optimal therapy has not yet been established. Combination of corticosteroids and cyclophosphamide is considered the best therapeutic approach available so far, but doses and duration of treatment need to be determined. Future research should focus on new anti-inflammatory or immunosuppressive agents.
Cholinergic Regulation of Morphine Release from Human White Blood Cells: Evidence for a Novel Nicotinic Receptor via Pharmacological and Micro Array Analysis
Authors: Zhu W. , Mantione K.J. , Kream R.M. , Cadet P. , Stefano G.B.
Published: January 2017

Recent work from our laboratory has demonstrated that human white blood cells make morphine and that substances of abuse, i.e. nicotine, alcohol and cocaine have the ability to release this endogenous substance, suggesting a common mechanism of action. We now demonstrate that the nicotinic process is more complex than formerly envisioned. The incorporation rate of 125I-labeled morphine into PMN and MN are 7.85±0.36%, 1.42±0.19%, respectfully, suggesting in MN this process is of low activity. Separate incubations of PMN with varying concentrations of nicotine or the nicotine agonist epibatidine resulted in a statistically significant enhancement of 125I-trace labeled morphine released into the extracellular medium. In order to ascertain the specificity of the nicotine stimulated morphine release the following experiments were performed. Co-incubation of hexamethonium dichloride (5 μg/ml and at 10 μg/ml), which preferentially blocks nicotinic receptors at autonomic ganglia, with nicotine, exerted a very weak inhibitory effect. Co-incubation of α-BuTx or atropine or chlorisondamine diiodide or dihydro-β-erythroidine hydrobromide, an α4β2 receptor antagonist, did not block nicotine induced morphine release alone or in combination, suggesting either the response was not specific or it was mediated by a novel nicotinic receptor. Human leukocyte total RNA isolated from whole blood were analyzed, using the Human Genome Survey microarray (Applied Biosystems), for cholinergic receptor expression. PMN nicotinic receptor gene expression was present and contained numerous variants (eight). The number of variants suggests that indeed a novel nicotinic receptor may be mediating this effect, while simultaneously demonstrating the significance of the cholinergic receptor expression in these immune cells.
TT Virus Infection: Role of Interferons, Interleukin-28 and 29, Cytokines and Antiviral Proteins
Authors: Garbuglia A.R. , Grasso F. , Donà M.G. , Mochi S. , Conti P. , De Lutiis M.A. , Giorgi C. , Iezzi T.
Published: January 2017

In 1997 a novel virus in the serum of a patient with acute post-transfusion hepatitis of non A-G etiology was identified. This agent was designed TT virus (TTV). It produces persistent viremia and no disease, but the mechanism of its persistence is poorly understood. In the present study mRNA expression of antiviral proteins as MxA, 2′5′ OAS, anti-apopotic protein, cytokines IL- 28, IL- 29 and IFN are examined in a subject affected by B lymphoma and positive for TTV DNA and RNA in this cellular subset, and in BJAB and Dohh2 cell lines.
Interleukin-3 Promotes the Expression of E-NPP3/CD203C on Human Blood Basophils in Healthy Subjects and in Patients with Birch Pollen Allergy
Authors: Hauswirth A.W. , Sonneck K. , Florian S.T. , Krauth M.-T. , Böhm A. , Sperr W.R. , Valenta R. , Schernthaner G.-H. , Printz D. , Fritsch G. , Bühring H.-J. , Valent P.
Published: January 2017

We recently identified the ectoenzyme CD203c as a novel basophil activation antigen that is up-regulated in response to FceRI cross-linkage. We investigated the effects of various interleukins (ILs) on expression of CD203c on blood basophils using an antibody against CD203c and flow cytometry. Of all cytokines tested, only IL-3 was found to upregulate expression of CD203c on basophils above baseline levels. The effects of IL-3 were dose- and time-dependent (EC50: 0.1–1 ng/ml) without differences observed between healthy and allergic donors. Whereas anti-IgE induced maximum upregulation of CD203c within 15 minutes, the IL-3-induced upregulation showed a maximum after 180 minutes. IgE-receptor cross-linking resulted in enhanced expression of both CD63 and CD203c, whereas IL-3 enhanced the levels of CD203c without promoting expression of CD63. The IL-3-induced upregulation of CD203c was also observed in highly enriched basophils and was counteracted by a blocking antibody against the alpha chain of the IL-3 receptor (CD123). The IL-3-induced upregulation of CD203c was also found to depend on the presence of calcium. To analyze signaling pathways involved in IL-3-induced upregulation of CD203c, pharmacologic inhibitors were applied. The PI3-kinase inhibitors, wortmannin and LY294002 counteracted the IL-3-induced expression of CD203c, whereas MEK- and PKC inhibitors showed no effects. In conclusion, IL-3 upregulates expression of CD203c on basophils through a specific receptor and via a PI3-kinase-dependent signaling-pathway. Compared to FceRI-mediated cell activation, IL-3-induced upregulation of CD203c is a late(r) event and is not accompanied by upregulation of CD63.
Tannic Acid Induces in vitro Acantholysis of Keratinocytes via IL-1α and TNF-α
Authors: Feliciani C. , Ruocco E. , Zampetti A. , Toto P. , Amerio P.A. , Tulli A. , Amerio P. , Ruocco V.
Published: January 2017

The mechanism of acantholysis in pemphigus vulgaris (PV) is an intriguing argument since several chemical mediators are implicated. We previously reported a central role for IL-1α and TNF-α, both able to regulate complement activation and plasminogen activators. Very little is known about what triggers the disease (drugs, viruses or food). In this study, we evaluate the molecular role of tannins in acantholysis. By HPLC chromatography we measured tannic acid (TA) and gallic acid (GA) in blister fluid of 4 groups of patients divided according to their dietary habits, including a regular diet, a diet rich in tannins, a diet free of tannins, and a group of pemphigus patients. Blister fluid was obtained from patients using a suction blister apparatus. We show that people with a diet rich in tannins have increased tannin metabolites (TA and GA) in the skin in respect to controls (tannin-rich diet: GA = 194.52±2.39 nmol/ml; TA = 348.28±1.4 nmol/ml versus tannin-Mediterranean diet: GA = 15.28±1.63 nmol/ml; TA = 22.81±1.68 nmol/ml). PV patients showed similar values to the Mediterranean diet population (PV patients: GA = 95.8±1.97 nmol/ml; TA = 199.09±4.15 nmol/ml versus Mediterranean diet: GA = 83.53±2.35 nmol/ml; TA = 195.1±2.50 nmol/ml). In an in vitro acantholysis system using TA and PV-IgG we show that TA 0.1 mM in NHEK culture is able to induce acantholysis. This effect was able to amplify the acantholytic action of PV-IgG in vitro. A blocking study using anti IL-1α and anti TNF-α antibodies showed a reduction in TA-induced acantholysis. Taken together, these results suggest that a diet rich in tannins could be a trigger in genetically predisposed patients. If these data are confirmed, a complementary diet poor in tannins may be useful in patients affected by PV.
In Melanoma Changes of Immature and Mature Dendritic Cell Expression Correlate with Tumor Thickness: An Immunohistochemical Study
Authors: Simonetti O. , Goteri G. , Lucarini G. , Rubini C. , Stramazzotti D. , Muzio L. Lo , Biagini G. , Offidani A.
Published: January 2017

Cells with a dendritic morphology and/or expression of dendritic cell (DC) markers have been repeatedly described in several human tumors, but the distribution and density of melanoma-associated DCs have not yet been reported. The aim of the present study is to analyze the density and topographical distribution of melanoma-associated DCs and their relation with CD3+, CD4+ and CD8+ T lymphocytes in forty cases of cutaneous human melanoma. In melanocytic tumours different pools of DCs were recognised in the epidermis and in the dermis, particularly in intimate relation with lymphocyte clusters inside the melanocytic proliferation, and more often at the edges of tumours. The number of Langerin-positive DCs showed an inverse correlation with tumour depth (correlation coefficient r= −0.59, P=0.0001) and was significantly lower in thick melanomas compared to thin and intermediate ones (P<0.0005). The density of CD83+ DCs was significantly lower in thick melanomas compared to thin and intermediate ones (P<0.009). A significant correlation was found between the density of the two DCs subsets (r=0.57, p<0.0001). The number of CD3+ lymphocytes was inversely correlated to the depth of infiltration (r=−0.596, P<0.0001): melanoma cases with II-III Clark level showed a higher T lymphocyte mean density compared to cases with IV-V Clark's level (P<0.0001). T lymphocyte density was significantly lower in thick melanomas compared to thin and intermediate melanomas (P<0.0005). In conclusion, our study indicates a progressive loss of DCs and T lymphocytes in the neoplastic progression of melanomas; further identification of the molecular pathways involved in the functional impairment of these immunitary cells may lead to new immunotherapeutic approaches for melanoma patients that would improve the clinical outcome of the patients.
Management and Triage of Women with Human Papillomavirus Infection in Follow-up for Low-Grade Cervical Disease: Association of HPV-DNA and RNA-Based Methods
Authors: Rosini S. , Zappacosta R. , Di Bonaventura G. , Caraceni D. , Pilla D. , Di Girolamo G. , Esposito A. , Orsini T. , Setta S. , Vizzino M. , Piccolomini M. , Piccolomini R.
Published: January 2017

Type-specific persistent infection with Human Papillomavirus (HPV) is a significant risk factor for the development of cervical diseases. Persistent infection could be further refined by a sequencing approach to detect early cervical lesions that are at high risk of developing an invasive squamous cervical cancer. The aim of the present study is to investigate the clinical utility of detecting mRNA transcripts of HPV oncogenes E6/E7 by using a Real-time NASBA technology (mRNA test) and to identify women with low-grade cytological disease but with an increased risk of developing high-grade cervical abnormalities or invasive squamous cervical cancer. Our preliminary results show that E6/E7 is detected in only a subset of HR-HPV-positive cases. Since viral persistence is considered to be the true precursor of neoplastic progression, only the detection of E6/E7 mRNA can identify the infection which is more likely to persist and induce neoplasia in future. For these reasons we believe that this test would be useful for the characterization of women with HR-HPV DNA positivity who should be effectively treated because at high-risk of developing a high grade cervical lesion or an invasive squamous cervical cancer.
Internalization of Non-Clustered Desmoglein 1 without Depletion of Desmoglein 1 from Adhesion Complexes in An Experimental Model of the Autoimmune Disease Pemphigus Foliaceus
Authors: Lanza A. , De Rosa A. , Femiano F. , Annese P. , Ruocco E. , Gombos F. , Lanza M. , Cirillo N.
Published: January 2017

Serum antibodies against desmoglein 1 (Dsg1) are known to induce the clinical and histological manifestations of pemphigus foliaceus (PF), autoimmune bullous disease targeting skin. The basic pathophysiological phenomenon of PF blistering is the disruption of epithelial integrity in the granular layer of the epidermis due to separation of keratinocytes from one another, or acantholysis. In this report we investigate the changes in subcellular distribution of Dsg1 in response to serum of patients with PF by using an in vitro model of PF. Immunofluorescence analysis on HaCaT cells indicates that non-clustered Dsg1 is markedly internalized after exposure to serum. However, binding of PF IgG to Dsg1-rich adhesion complexes (desmosomes) does not cause disruption of such structures nor depletion of clustered Dsg1, as revealed by colocalization of PF IgG and Dsg1 in a punctate staining on cell membrane 24 hours after treatment. Furthermore, morphological studies demonstrate that the dramatic alterations induced by PF sera are not the result of apoptotic programs. Taken together, our data strongly suggest that anti-Dsg1 antibodies from PF serum could cause the internalization of non-clustered Dsg1 and perturb the formation of new desmosomes but not directly disrupt Dsg1-containing junctions when stable contacts are already formed.
A Novel Unconventional Antigen MPD5 Elicits Anti-Tumor Humoral Immune Responses in a Subset of Patients with Polycythemia Vera
Authors: Xiong Z. , Liu E. , Yan Y. , Silver R.T. , Yang F. , Chen I.H. , Hodge I. , Verstovsek S. , Segura F.J. , Wang H. , Prchal J. , Yang X.-F.
Published: January 2017

In an effort to define the antigenic mechanism that contributes to beneficial therapeutic outcome in patients with polycythemia vera (PV), we screened a human testis cDNA library with serological cloning derived from sera of three PV patients who had undergone therapeutic-induced remission. As a result, we identified a novel antigen, MPD5, which belongs to the group of cryptic antigens with unconventional genomic intron/exon structure. Moreover, MPD5 elicited IgG antibody responses in a subset of PV patients who had benefited from a variety of therapies—including IFN-α, Hydroxyurea, Imatinib mesylate, Anagrelide, and phlebotomy—but not in untreated PV patients or healthy donors, suggesting that MPD5 is a PV-associated, therapy-related antigen. In the granulocytes of PV patients who are responsive to therapy, upregulated MPD5 expression may serve to enhance immune responses. These findings provide new insight into the mechanism underlying regulation of the self-antigen repertoire that elicits anti-tumor immune responses in patients with myeloproliferative diseases, indicating the potential of these self-antigens as targets of novel immunotherapy.
A Case of Human Polyomavirus BK Infection in a Patient Affected by Late Stage Prostate Cancer: Could Viral Infection Be Correlated with Cancer Progression?
Authors: Fioriti D , Russo G. , Mischitelli M. , Anzivino E. , Bellizzi A. , Di Monaco F. , Di Silverio F. , Giordano A. , Chiarini F. , Pietropaolo V.
Published: January 2017

The basic molecular mechanisms regulating prostate cancer (PCA) development and progression are very poorly understood. Different tumor suppressor genes are implicated in PCA. In particular, since the mutation rate of the p53 gene is also low, researchers have speculated that an infectious agent might play an important role in PCA. Polyomaviruses are candidates for this agent. We selected a patient with a diagnosis of PCA and underwent radical prostatectomy, to investigate the presence of Polyomavirus BK (BKV) sequences (urine and neoplastic tissues) and the mutation pattern of p53 gene. The results obtained showed the presence of BKV DNA and of p53 gene mutations in exons 6, 8 and 9. We speculate that BKV might contribute to cellular transformation process, triggered possibly by p53 gene mutations.
Recombinant Allergens Promote Expression of Aminopeptidase-N (CD13) on Basophils in Allergic Patients
Authors: Sonneck K. , Baumgartner C. , Rebuzzi L. , Marth K. , Chen K.W. , Hauswirth A.W. , Florian S. , Vrtala S. , Bühring H.J. , Valenta R. , Valent P.
Published: January 2017

IgE-dependent activation of basophils is associated with upregulation of several surface molecules. We recently identified the surface enzyme aminopeptidase N (CD13) as a novel activation antigen on human basophils. In the present study, we asked whether CD13 can be employed as a novel marker of allergen-induced activation of basophils in allergic individuals. Patients allergic to major allergens from grass pollen (Phl p 1, Phl p 5), birch pollen (Bet v 1), or house dust mites (Der p 2), were examined. Blood basophils were exposed to various concentrations of recombinant allergens for 15 minutes, and examined for expression of CD13 by multicolor flow cytometry. The allergen-induced upregulation of CD13 was compared with allergen-dependent increases in expression of CD63 and CD203c. Exposure to recombinant allergens resulted in an increase in expression of CD13 on basophils in all sensitized individuals, whereas no increase in CD13 was seen in healthy controls. The effects of the recombinant allergens on CD13-expression were dose- and time-dependent, were not observed in the absence of extracellular calcium, and were counteracted by preincubation of basophils with the PI3-kinase-targeting drugs staurosporin and LY294002. There was a good correlation between allergen-induced upregulation of CD13, CD63, and CD203c on basophils. In aggregate, our data show that recombinant allergens promote expression of CD13 on basophils in sensitized individuals. The functional significance and diagnostic implications of this observation remain to be determined.
Evaluation of Neutrophil CD64 Expression and Procalcitonin as Useful Markers in Early Diagnosis of Sepsis
Authors: Cardelli P. , Ferraironi M. , Amodeo R. , Tabacco F. , De Blasi R.A. , Nicoletti M. , Sessa R. , Petrucca A. , Costante A. , Cipriani P.
Published: January 2017

Quantitation of neutrophil CD64 expression and procalcitonin (PCT) levels in blood samples have been recently proposed as useful tools for early detection of sepsis. To determine the usefulness of these tests, we analyzed blood samples of 112 patients, admitted to an intensive care unit (ICU), presenting clinical symptoms of sepsis, as well as of 50 healthy controls. At the end of the study, a retrospective analysis showed that only 52 of the 112 ICU-patients presented a real sepsis (positive blood culture). The results obtained indicated that of the 52 patients with sepsis, 50 and 49 presented levels of neutrophil CD64 expression ≥ 2398 molecules per cell (cut-off determined by receiver operator characteristic analysis) and PCT levels > 0.5 ng/ml (cut-off suggested by the manufacturer), respectively. However, the neutrophil CD64 test showed higher specificity in detecting sepsis since 5 out of the 60 ICU-patients without sepsis (negative blood culture), presented CD64 expression levels ≥ 2398 molecules per cell, PCT levels ≥ 0.5 ng/ml were shown in 27 patients. Moreover, while none of the 50 healthy controls presented a neutrophil CD64 level higher than the cut-off value, 5 patients presented PCT levels ≥ 0.5 ng/ml. In conclusion, our data seem to indicate that the quantitation of CD64 expression could be taken into consideration as a sensitive and specific test for early diagnosis of sepsis.
Omeprazole Induces Apoptosis in Normal Human Polymorphonuclear Leucocytes
Authors: Capodicasa E. , Cornacchione P. , Natalini B. , Bartoli A. , Coaccioli S. , Marconi P. , Scaringi L.
Published: January 2017

We investigated in vitro apoptosis in human polymorphonuclear neutrophils (PMN) induced by omeprazole. This drug, both in the native (OM) and acidified (OM-HCl) form, is a potent inducer of PMN apoptosis. The effect is time- and dose-dependent. OM-HCl is more efficient than OM in inducing PMN apoptosis. In fact, after 24 h incubation in vitro at 1×10 −4M OM-HCl induces apoptosis in 70% of the cell population compared to 37% induced by OM. Apoptosis induced by both forms of the drug is caspase dependent being significantly reduced by pretreating cells with the caspase 3 inhibitor (DEVDH-CHO). However, some differences in the apoptosis mechanisms between the two forms of the drug seem to exist because PMN treatment with the specific caspase 8 inhibitor (Z-IETD-FMK) only blocks OM-HCl mediated apoptosis. We observed cleavage of caspase 8 only in the cells incubated with OM-HCl while the executioner caspase 3 was activated with both forms of the drug. Furthermore, pretreatment with GM-CSF, a known activator of intracellular survival pathways in PMN, partially protected cells from OM-HCl induced apoptosis but did not contrast the apoptotic effect of OM. Cysteine cathepsin proteases also seem involved in the apoptotic mechanism of both drug forms since the specific inhibitor E64d gave a significant protection. To verify if OM-HCl induced apoptosis was dependent on the sulfenamide bound with the cell sulfhydryl groups we used molecules with thiol groups such as β-mercaptoethanol (β-ME) and reduced glutathione (GSH). Reactions of OM-HCl with cellular sulfhydryl groups are strongly involved in both the triggering and evolving phase of the apoptotic mechanism since significant protection from apoptosis was obtained when PMN were pretreated for 1h with β-ME (lipid-permeable) or GSH (lipid-impermeable). These results show that OM and OM-HCl induce apoptosis in human PMN and suggest that the second binds the sulfhydryl groups, present on the cell membrane, to then penetrate the cell thus causing a further significant increase in apoptosis. OM-induced PMN apoptosis during the treatment of gastric inflammatory disease could be an advantage for the resolution of the phlogosis state. However, this aspect should be further elucidated to assess the optimal therapeutical regimen for gastric diseases which are related to infective agents.
How Do Rat Cortical Cells Cultured with Aluminum Die: Necrosis or Apoptosis?
Authors: Zhang Q.L. , Boscolo P. , Niu P.Y. , Wang F. , Shi Y.T. , Zhang L. , Wang L.P. , Wang J. , Di Gioacchino M. , Conti P. , Li Q.Y. , Niu Q.
Published: January 2017

Aluminum (Al) exposure has been implicated as the cause of neural cells loss in several neurodegenerative diseases. Therefore, defining the mechanism of neural cell death in Al toxicity and degenerative diseases might lead to the development of therapeutic agents which promote neural cell survival. Furthermore, knowledge of cell death pathways might facilitate the discovery of treatments for neurodegeneration. However, the death mode of neural cells triggered by Al has not been firmly established. The present study focuses on understanding the pathway of cells death in cultured cortical cells treated with Al. Primary neurons cultured alone, astrocytes cultured alone, and neuron/astrocyte co-cultures obtained from newborn rats were incubated with Al at the concentrations of 0, 0.5,1.0, or 2.0 mM for 72 h. Morphological changes were observed with an inverted phase microscope, a fluorescent microscope, and an electron microscope. Simultaneously, the rate of apoptosis was quantified with flow cytometry. Morphological characteristics of apoptosis such as cell shrinkage, aggregation and fragmentation of chromatin, membrane buds, and formation of membrane-bound apoptotic bodies were observed in Al-treated neurons, while none of these characteristics were found in Al-treated astrocytes. Quantitative results of apoptotic rates detected with flow cytometry indicated a typical apoptosis progression in neurons at various dosages. A concentration-dependent relationship between Al concentration and apoptotic rates confirmed that apoptosis is the prominent cause of cell death in primary cultured neurons, even at a concentration lower than 2 mM. Both necrosis and apoptosis are evident in neuron/astrocyte co-cultures, but the intensity of apoptosis is much less compared with that of neurons, suggesting that astrocytes may be especially important for neuronal survival in the presence of Al.
Activity and Expression of Nitric Oxide Synthase in Rat Bladder after Sacral Neuromodulation
Authors: Minardi D. , Ghiselli R. , Lucarini G. , Mocchegiani F. , Filosa A. , Zizzi A. , Simonetti O. , Orlando F. , Pelliccioni G. , Parri G. , Saba V. , Muzio L. Lo , Biagini G. , Montironi R. , Muzzonigro G.
Published: January 2017

The aim of our study is to investigate the effects of chronic sacral neuromodulation on Nitric Oxide (NO) metabolism in the rat bladder. 26 female Sprangue-Dawley rats were considered: group I, normal control rats; group II, a sham treatment, in whom catheters for electrical stimulation were placed in the S1 foramen bilaterally and left in place for 21 days, without performing neuromodulation; group III in whom electrical sacral neuromodulation was performed for 21 days. Finally a cystectomy was performed and the bladder biopsy specimens were sent for immunostaining with n-NOS and i-NOS. Morphological and immunohistochemical analysis was carried out, and evaluated in urothelial cells, endothelial cells and muscle fibers of the muscularis propria. Differences between the 3 groups were analyzed by Student Newman-Keuls test. We could observe that urothelial and endothelial i-NOS (37.00±4.69 and 59.00±7.42 respectively) and urothelial n-NOS (36.80±7.85) expression are significantly increased in neuromodulated rats, compared to groups 1 and 2 (p < 0.005). In conclusion, the increase of i-NOS expression on endothelial cells after sacral neuromodulation could be in some way related to angiogenetic responses in the microvascular structures; the increase of n-NOS and i-NOS expression on urothelial cells can suggest that NO is able to influence the plasticity of bladder response, inducing the release of messengers within the urothelium. This study can therefore improve our understanding of the mechanisms of sacral neuromodulation on chronic bladder dysfunction; further studies will need to better demonstrate the role of angiogenesis in the bladder after sacral neuromodulation and to investigate the effects of neuromodulation in rats with chronically induced bladder dysfunction.
Role of Fosfomycin Tromethamine in Modulating Non-Specific Defence Mechanisms in Chronic Uremic Patients towards ESBL-Producing Escherichia Coli
Authors: Tullio V. , Cuffini A.M. , Banche G. , Mandras N. , Allizond V. , Roana J. , Giacchino F. , Bonello F. , Ungheri D. , Carlone N.A.
Published: January 2017

Antimicrobial agents and polymorphonuclear cells (PMNs) have the potential to interact in such a way that improve the therapy for infectious diseases. In immunocompromised patients highly susceptible to microbial infections with high morbidity and mortality, several metabolic and functional alterations in PMNs, mostly related to microbicidal activity, are observed. Therefore, the antibiotic of choice should have a good antimicrobial effect without impairing host defences. The aim of this study is to evaluate in vitro effects of sub-inhibiting fosfomycin tromethamine (FT) concentrations on the primary functions of PMNs from healthy subjects and immunocompromised patients (haemodialysed and renal transplant recipients), against an ESBL-producing Escherichia coli, the most common aetiological agent in urinary tract infections (UTIs). FT is considered a first line drug in the eradication of UTIs due to its appropriate antimicrobial spectrum, oral bioavailability and minimal risk of microbial resistance. Our results provide evidence that FT is able to induce enhancement of the depressed phagocytic response of PMNs from patients on chronic haemodialysis and from renal transplant recipients, restoring their primary functions in vitro against ESBL-producing E. coll All these data permit the conclusion that uremic-infected patients might additionally benefit from the immunomodulating properties of FT.
Epithelial Lining Fluid Neutrophil-Gelatinase-Associated Lipocalin Levels in Premature Newborns with Bronchopulmonary Dysplasia and Patency of Ductus Arteriosus
Authors: Capoluongo E. , Vento G. , Lulli P. , Di Stasio E. , Porzio S. , Vendettuoli V. , Tana M. , Tirone C. , Romagnoli C. , Zuppi C. , Ameglio F.
Published: January 2017

Patency of the ductus arteriosus (PDA) and bronchopulmonary dysplasia (BPD) development represent severe affections for premature newborns, therefore the research of early markers for these two conditions is really important. The aim of this study is to analyze epithelial lining fluid (ELF) Neutrophil-gelatinase-associated lipocalin (NGAL) levels for prediction of lung injury or possible involvement of this molecule in PDA. Only scarce and contrasting results have previously been published in this field. In contrast, this molecule, included in a large macromolecular complex together with matrix metalloproteinase-9 (MMP-9), is considered an acceptable marker of infectious/inflammatory processes, cancer monitoring and induction of apoptotic pathway. NGAL was detected in 28 pre-term newborns by means of a commercially available kit in bronchoalveolar lavage fluid (BALF). The results have been corrected to ELF levels, by the urea method, to eliminate bias due to BALF collection. ELF NGAL levels were found significantly increased both in infants developing BPD or in those affected by PDA. By means of multivariate logistic regression analysis the significances were confirmed after adjusting for possible interfering variables such as gestational age and concomitant presence of both PDA and BPD. Our results stress the involvement of NGAL in the mechanisms leading to BPD and also suggest a possible association with PDA, which is often linked to prematurity and BPD development, probably due to the involvement of inflammatory and angiogenetic processes in both pathologies.
Intravitreal Micronized Triamcinolone versus Triamcinolone Acetonide: A Clinical and Morphological Comparative Study
Authors: Salvolini E. , Neri P. , Orciani M. , Di Primio R. , Giovannini A.
Published: January 2017

Nowadays many authors suggest the use of intravitreal triamcinolone acetonide (TA) for the treatment of vitreoretinal diseases, although it can be associated with a high risk of local toxicity. In order to develop a safer injection for clinical use, the purpose of our study is to evaluate the in situ safety of two different triamcinolone preparations, a commercially available TA and a micronized triamcinolone. The experiments were performed on 18 adult male age-matched New Zealand rabbits. The clinical examination included funduscopy with an indirect ophthalmoscope and intraocular pressure (IOP) measurement. At the end of the clinical observations, the animals were sacrified and the eyes enucleated and processed for the morphological evaluation. In our study the main side effect observed was the IOP elevation in the group injected with triamcinolone acetonide. In addition, in the TA-injected group, one eye was enucleated following an endophthalmitis. Our study highlights that doses as low as 4 mg of triamcinolone acetonide injected into the rabbit vitreous may have a local toxic effect in terms of IOP elevation, endophthalmitis occurrence and changes in the retinal morphology. In contrast, the micronized triamcinolone injection shows a less toxic effect in situ, thus suggesting the alternative use of this more reliable preparation which seems to be safer for a clinical use.