Aspetti processuali del Regolamento n. 1/2003/Ce

The recent reform in European antitrust enforcement is embodied in Regolation n. 1/2003/ Ce and related Communications. Since 2004 when it came into force, some crytical assessments can already be made. The work starts from some technical analysis of the reform, under a procedural perspective, to assess the proceedings’ real impact on parties’ rights and to criticize its limits. Decentralisation has brought about more complicacies, since community procedural systems are not harmonized, neither in their administrative rules, nor in their civil proceedings, which are all involved in the European antitrust network. Therefore, antitrust proceedings end un as being more jurisdictional in their effects than in their guarentees, which is a flaw to be mended by legislators. National laws shoud be harmonized, community law should be clarified and the system should turn more honestly towards a rationalized jurisdiction-cented mechanism. Otherwise, parties defense rights and the overall efficiency are put into doubt. Italy is a good exemple of how many colmlicacies can outburst from national procedures and national decentralised application. An uncertain pattern of judicial control, together with unclear relationships among the institutions to cooperate in the antitrust network can produce more problems than they aim to solve. As to the private enforcement, Regulation n.1 does not even attempt to give precise regulation to this underdeveloped sector. A continual comparison with U.S. system has brought the Commission to become aware both of the risks and of the advanteges of an increased civil antitrust litigation in fronto of national judges. In order to substain a larger development of this parallel and, presently, difficult way of judicial compensation, it is presently ongoing a consultation among states to find suitable incentives to make private enforcement more appealing and effective. The solution to this lack of private litigation is not to be sought in Regulation n. 1 which calls into action national legislators and proceedures to implement further improvements. As a conclusion, Regulation n. 1 is the outpost of an ambitious community design to create an efficient control mechanism over antitrust violations. It focuses on Commission proceedings, powers and sanctions in order to establish deterrence, then it highlights civil litigation perspectives and it involves directly states into antitrust application. It seems that more could be done to technically shape administrative proceedings in a more jurisdictionally oriented form, then to clarify respective roles and coordination mecanisms in order to prevent difficulties easy to forsee. Some of jurisprudential suggestions have been accepted, but much more is left to be done in the future to improve european antitrust enforcement system.

Präparation, Charakterisierung und elektronische Eigenschaften von dünnen Schichten des Hochtemperatursupraleiters HgReBa 2 Ca n-1 Cu n O y

Ziel dieser Arbeit war die Pr"{a}paration, Charakterisierung und Untersuchung der elektronischen Eigenschaften von d"{u}nnen Schichten des Hochtemperatursupraleiters HgReBa$_{2}$Ca$_{n-1}$Cu$_{n}$O$_{y}$, die mittels gepulster Laser-Deposition hergestellt wurden. Die HgRe1212-Filme zeigen in der AC-Suszeptibilit"{a}t einen scharfen "{U}bergang in die supraleitende Phase bei 124 K mit einer "{U}bergangsbreite von 2 K. Die resistiven "{U}berg"{a}nge der Proben wurden mit zunehmender St"{a}rke des externen Magnetfeldes breiter. Aus der Steigung der Arrheniusplots konnte die Aktivierungsenergie f"{u}r verschiedene Feldst"{a}rken bestimmt werden. Weiterhin wurde die Winkelabh"{a}ngigkeit des Depinning-Feldes $B_{dp}(theta)$ der Filme gemessen. Hieraus wurde ein Anisotropiewert von $gamma$ = 7.7 bei 105 K ermittelt. Dies ist relevant, um den f"{u}r Anwendungen wichtigen Bereich im $T$-$B$-$theta$-Phasenraum des Materials absch"{a}tzen zu k"{o}nnen. Die kritische Stromdichte $J_{c}$ der d"{u}nnen Filme aus HgRe-1212 wurde mit Hilfe eines SQUID-Magnetometers gemessen. Die entsprechenden $M$-$H$ Kurven bzw. das magnetische Moment dieser Filme wurde f"{u}r einen weiten Temperatur- und Feldbereich mit einem magnetischen Feld senkrecht zum Film aufgenommen. F"{u}r einen HgRe-1212-Film konnte bei 5 K eine kritische Stromdichte von 1.2 x 10$^{7}$ A/cm$^{2}$ und etwa 2 x 10$^{6}$ A/cm$^{2}$ bei 77 K ermittelt werden. Es wurde die Magnetfeld- und die Temperaturabh"{a}ngigkeit des Hall-Effekts im normalleitenden und im Mischzustand in Magnetfeldern senkrecht zur $ab$-Ebene bis zu 12 T gemessen. Oberhalb der kritischen Temperatur $T_{c}$ steigt der longitudinale spezifische Widerstand $rho_{xx}$ linear mit der Temperatur, w"{a}hrend der spezifische Hall-Widerstand $rho_{yx}$ sich umgekehrt proportional zur Temperatur "{a}ndert. In der N"{a}he von $T_{c}$ und in Feldern kleiner als 3 T wurde eine doppelte Vorzeichen"{a}nderung des spezifischen Hall-Widerstandes beobachtet. Der Hall-Winkel im Normalzustand, cot $theta_{H}= alpha T^{2} + beta$, folgt einer universellen $textit{T }^{2}$-Abh"{a}ngigkeit in allen magnetischen Feldern. In der N"{a}he des Nullwiderstand-Zustandes h"{a}ngt der spezifische Hall-Widerstand $rho_{yx}$ "{u}ber ein Potenzgesetz mit dem longitudinalen Widerstand $rho_{xx}$ zusammen. Das Skalenverhalten zwischen $rho_{yx}$ und $rho_{xx}$ weist eine starke Feld-Abh"{a}ngigkeit auf. Der Skalenexponent $beta$ in der Gleichung $rho_{yx}$ =A $rho_{xx}^{beta}$ steigt von 1.0 bis 1.7, w"{a}hrend das Feld von 1.0 bis 12 T zunimmt.

Die Beeinflussung einer Th2-Zell-vermittelten Immunantwort durch Interleukin-1-alpha am Beispiel des murinen allergischen Asthmas

Dass durch IL-1α die Th1-vermittelte Immunreaktion der kutanen Leishmaniasis beeinflusst werden kann, konnte unsere Arbeitsgruppe bereits zeigen. Daran anknüpfend war das Ziel meiner Dissertation zu prüfen, ob sich diese Erkenntnisse im Modell des murinen allergischen Asthmas reproduzieren lassen, auch im Sinne eines zukünftigen therapeutischen Nutzens für die Behandlung dieser epidemiologisch hochrelevanten Erkrankung. Daneben sollte die Verwendung der gesamten murinen Lunge als Quelle für Untersuchungsmaterial erschlossen werden. Zu diesem Zweck wurden bei BALB/c Mäusen ein (OVA)/Alum-induziertes allergisches Asthma in Gegenwart oder Abwesenheit von IL-1α generiert. Anschließend wurde eine broncheoalveoläre Lavage (BAL) durchgeführt, bzw. die komplette linke Lunge gewonnen und prozessiert. Der Einfluss von IL-1α auf den Phänotyp der Th2-vermittelten Immunantwort ließ sich auf zytomorphologischer, durchflusszytometrischer und Zytokin-Ebene nachweisen. So zeigte ein Teil unserer Ergebnisse, dass nach der frühen Gabe eine Tendenz zur Verlagerung des Gewichtes der Abwehrreaktion von Th2 in Richtung Th1 besteht. Ebenso fanden sich Hinweise für eine relative Augmentation der Th2-Antwort durch den Einsatz von IL-1α zu einem späteren Zeitpunkt. Eine absolute Verstärkung der Th2-Reaktion auf OVA durch IL-1α konnten wir nicht messen. Hier scheint mit OVA allein schon eine maximale Ausprägung erreicht zu sein. Neben den Th1/Th2-Effekten wurden auch einige gegenläufige Beobachtungen gemacht, welche nicht a priori durch das Th1/Th2-Paradigma zu erklären sind, sondern den Einfluß von IL-1α auf andere Systeme belegen, wie z. B. die Wirkung auf CCL28 und regulatorische T-Zellen. Für die Gewinnung von inflammatorischen Zellen aus der kompletten Lunge und deren weitere Untersuchung konnten wir eine Methode entwickeln und standardisieren, welche relativ einfach in der Durchführung ist und zuverlässig eine im Verhältnis zur BAL hohe Zellzahl liefert, was wiederum ein breites Spektrum an weiteren Untersuchungen erlaubt. Durch den Vorgang der mechanischen und enzymatischen Prozessierung scheinen die funktionellen Eigenschaften der Zellen nicht wesentlich beeinträchtigt zu sein. Der Einsatz von IL-1α resultierte letztendlich in einem Mischbild an hervorgerufenen Veränderungen und es bedarf noch weiterer Studien, um die unterschiedlichen induzierten Mechanismen sauber voneinander zu trennen und den therapeutischen Nutzen von IL-1α im allergischen Asthma zu evaluieren.

Studio della patogenesi di HIV-1 nel compartimento osseo: effetti della terapia antiretrovirale convenzionale e nuovi approcci all'eradicazione

L’obiettivo della tesi è studiare il virus HIV-1 in relazione alle alterazioni sistemiche, riscontrate nel paziente HIV-infetto, in particolare alterazioni a carico del sistema scheletrico, indotte dal virus o dall’azione dei farmaci utilizzati nella terapia antiretrovirale (HAART). L’incidenza dell’osteoporosi nei pazienti HIV-positivi è drammaticamente elevata rispetto alla popolazione sana. Studi clinici hanno evidenziato come alcuni farmaci, ad esempio inibitori della proteasi virale, portino alla compromissione dell’omeostasi ossea, con aumento del rischio fratturativo. Il nostro studio prevede un follow-up di 12 mesi dall’inizio della HAART in una coorte di pazienti naïve, monitorando diversi markers ossei. I risultati ottenuti mostrano un incremento dei markers metabolici del turnover osseo, confermando l’impatto della HAART sull’omeostasi ossea. Successivamente abbiamo focalizzato la nostra attenzione sugli osteoblasti, il citotipo che regola la sintesi di nuova matrice ossea. Gli esperimenti condotti sulla linea HOBIT mettono in evidenza come il trattamento, in particolare con inibitori della proteasi, porti ad apoptosi nel caso in cui vi sia una concentrazione di farmaco maggiore di quella fisiologica. Tuttavia, anche concentrazioni fisiologiche di farmaci possono regolare negativamente alcuni marker ossei, come ALP e osteocalcina. Infine esiste la problematica dell’eradicazione di HIV-1 dai reservoirs virali. La HAART riesce a controllare i livelli viremici, ciononostante diversi studi propongono alcuni citotipi come potenziali reservoir di infezione, vanificando l’effetto della terapia. Abbiamo, perciò, sviluppato un nuovo approccio molecolare all’eradicazione: sfruttare l’enzima virale integrasi per riconoscere in modo selettivo le sequenze LTR virali per colpire il virus integrato. Fondendo integrasi e l’endonucleasi FokI, abbiamo generato diversi cloni. Questi sono stati transfettati stabilmente in cellule Jurkat, suscettibili all’infezione. Una volta infettate, abbiamo ottenuto una significativa riduzione dei markers di infezione. Successivamente la transfezione nella linea linfoblastica 8E5/LAV, che porta integrata nel genoma una copia di HIV, ha dato risultati molto incoraggianti, come la forte riduzione del DNA virale integrato.

Growth hormone (GH) deficiency type II: a novel GH-1 gene mutation (GH-R178H) affecting secretion and action

Context and Objective: Main features of the autosomal dominant form of GH deficiency (IGHD II) include markedly reduced secretion of GH combined with low concentrations of IGF-I leading to short stature. Design, Setting, and Patients: A female patient presented with short stature (height -6.0 sd score) and a delayed bone age of 2 yr at the chronological age of 5 yr. Later, at the age of 9 yr, GHD was confirmed by standard GH provocation test, which revealed subnormal concentrations of GH and a very low IGF-I. Genetic analysis of the GH-1 gene revealed the presence of a heterozygous R178H mutation. Interventions and Results: AtT-20 cells coexpressing both wt-GH and GH-R178H showed a reduced GH secretion after forskolin stimulation compared with the cells expressing only wt-GH, supporting the diagnosis of IGHD II. Because reduced GH concentrations found in the circulation of our untreated patient could not totally explain her severe short stature, functional characterization of the GH-R178H performed by studies of GH receptor binding and activation of the Janus kinase-2/signal transducer and activator of transcription-5 pathway revealed a reduced binding affinity of GH-R178H for GH receptor and signaling compared with the wt-GH. Conclusion: This is the first report of a patient suffering from short stature caused by a GH-1 gene alteration affecting not only GH secretion (IGHD II) but also GH binding and signaling, highlighting the necessity of functional analysis of any GH variant, even in the alleged situation of IGHD II.

Psychogenic aphonia: no fixation even after a lengthy period of aphonia

Although psychogenic aphonia is considered to be a conversion disorder, aphonic patients are primarily treated symptomatically. This is because it is considered of overriding importance to elicit a voice quickly to avoid fixation of the aphonia. The aim of this study was to show that, for patients exhibiting the symptom of voicelessness, not eliciting the voice immediately will not lead to a permanent aphonia.

Expression patterns of Bmi-1 and p16 significantly correlate with overall, disease-specific, and recurrence-free survival in oropharyngeal squamous cell carcinoma

BACKGROUND: The objective of this study was to link expression patterns of B-cell-specific Moloney murine leukemia virus integration site 1 (Bmi-1) and p16 to patient outcome (recurrence and survival) in a cohort of 252 patients with oral and oropharyngeal squamous cell cancer (OSCC). METHODS: Expression levels of Bmi-1 and p16 in samples from 252 patients with OSCC were evaluated immunohistochemically using the tissue microarray method. Staining intensity was determined by calculating an intensity reactivity score (IRS). Staining intensity and the localization of expression within tumor cells (nuclear or cytoplasmic) were correlated with overall, disease-specific, and recurrence-free survival. RESULTS: The majority of cancers were localized in the oropharynx (61.1%). In univariate analysis, patients who had OSCC and strong Bmi-1 expression (IRS >10) had worse outcomes compared with patients who had low and moderate Bmi-1 expression (P = .008; hazard ratio [HR], 1.82; 95% confidence interval [CI], 1.167-2.838); this correlation was also observed for atypical cytoplasmic Bmi-1 expression (P = .001; HR, 2.164; 95% CI, 1.389-3.371) and for negative p16 expression (P < .001; HR, 0.292; 95% CI, 0.178-0.477). The combination of both markers, as anticipated, had an even stronger correlation with overall survival (P < .001; HR, 8.485; 95% CI, 4.237-16.994). Multivariate analysis demonstrated significant results for patients with oropharyngeal cancers, but not for patients with oral cavity tumors: Tumor classification (P = .011; HR, 1.838; 95%CI, 1.146-2.947) and the combined marker expression patterns (P < .001; HR, 6.254; 95% CI, 2.869-13.635) were correlated with overall survival, disease-specific survival (tumor classification: P = .002; HR, 2.807; 95% CI, 1.477-5.334; combined markers: P = .002; HR, 5.386; 95% CI, 1.850-15.679), and the combined markers also were correlated with recurrence-free survival (P = .001; HR, 8.943; 95% CI, 2.562-31.220). CONCLUSIONS: Cytoplasmic Bmi-1 expression, an absence of p16 expression, and especially the combination of those 2 predictive markers were correlated negatively with disease-specific and recurrence-free survival in patients with oropharyngeal cancer. Therefore, the current results indicate that these may be applicable as predictive markers in combination with other factors to select patients for more aggressive treatment and follow-up. Cancer 2011;. © 2011 American Cancer Society.

An Actor-Critic based controller for glucose regulation in type 1 diabetes

A novel adaptive approach for glucose control in individuals with type 1 diabetes under sensor-augmented pump therapy is proposed. The controller, is based on Actor-Critic (AC) learning and is inspired by the principles of reinforcement learning and optimal control theory. The main characteristics of the proposed controller are (i) simultaneous adjustment of both the insulin basal rate and the bolus dose, (ii) initialization based on clinical procedures, and (iii) real-time personalization. The effectiveness of the proposed algorithm in terms of glycemic control has been investigated in silico in adults, adolescents and children under open-loop and closed-loop approaches, using announced meals with uncertainties in the order of ±25% in the estimation of carbohydrates. The results show that glucose regulation is efficient in all three groups of patients, even with uncertainties in the level of carbohydrates in the meal. The percentages in the A+B zones of the Control Variability Grid Analysis (CVGA) were 100% for adults, and 93% for both adolescents and children. The AC based controller seems to be a promising approach for the automatic adjustment of insulin infusion in order to improve glycemic control. After optimization of the algorithm, the controller will be tested in a clinical trial.

An unusual patient with hypercalciuria, recurrent nephrolithiasis, hypomagnesemia and G227R mutation of Paracellin-1. An unusual patient with hypercalciuria and hypomagnesemia unresponsive to thiazide diuretics

A 19-year-old female patient with hypercalciuria and recurrent nephrolithiasis/urinary tract infection unresponsive to thiazide type diuretics is presented. The patient first experienced nephrolithiasis at the age of 4 years. Afterwards, recurrent passages of stones and urinary tract infection occurred. On diagnostic evaluation at the age of 19 years, she also had hypocitraturia and hypomagnesemia. Her serum calcium concentrations were near the lower limit of normal (8.5-8.8 mg/dl; normal range: 8.5-10.5), her serum magnesium concentrations were 1.15-1.24 mg/dl (normal range: 1.4-2.5) and urinary calcium excretion was 900 mg/24 h. PTH concentrations were increased (110-156 pg/ml; normal range: 10-65). We tried to treat the patient with hydrochlorothiazide at a dose of 50 mg/day. During treatment with thiazide diuretics, PTH concentration remained high and the patient had recurrent urinary tract infections and passages of stones. Serum magnesium concentration did not normalize even under the parenteral magnesium infusion. Her mother had a history of nephrolithiasis 20 years ago. Severe hypomagnesemia in association with hypercalciuria/urinary stones is reported as a rare autosomal recessive disorder caused by impaired reabsorption of magnesium and calcium in the thick assending limp of Henle's loop. Recent studies showed that mutations in the CLDN16 gene encoding paracellin-1 cause the disorder. In exon 4, a homozygous nucleotide exchange (G679C) was identified for the patient. This results in a point mutation at position Glycine227, which is replaced by an Arginine residue (G227R). The mother was heterozygous for this mutation. G227 is located in the fourth transmembrane domain and is highly conserved in the claudin gene family. This case indicates the pathogenetic role of paracellin-1 mutation in familial hypomagnesemia with hypercalciuria and nephrocalcinosis and further underlines the risk of stone formation in heterozygous mutation carriers.

The cytidine deaminases AID and APOBEC-1 exhibit distinct functional properties in a novel yeast selectable system

Activation-induced cytidine deaminase (AID) is indispensable for immunoglobulin maturation by somatic hypermutations and class switch recombination and is supposed to deaminate cytidines in DNA, while its homolog APOBEC-1 edits apolipoprotein (apo) B mRNA by cytidine deamination. We studied the editing activity of APOBEC-1 and AID in yeast using the selectable marker Gal4 linked to its specific inhibitor protein Gal80 via an apo B cassette (Gal4-C) or via the variable region of a mouse immunoglobulin heavy chain gene (Gal4-VH). Expression of APOBEC-1 induced C to U editing in up to 15% of the Gal4-C transcripts, while AID was inactive in this reaction even in the presence of the APOBEC-1 complementation factor. After expression of APOBEC-1 as well as AID approximately 10(-3) of yeast cells survived low stringency selection and expressed beta-galactosidase. Neither AID nor APOBEC-1 mutated the VH sequence of Gal4-VH, and consequently the yeast colonies did not escape high stringent selection. AID, however, induced frequent plasmid recombinations that were only rarely observed with APOBEC-1. In conclusion, AID cannot substitute APOBEC-1 to edit the apo B mRNA, and the expression of AID in yeast is not sufficient for the generation of point mutations in a highly transcribed Gal4-VH sequence. Cofactors for AID induced somatic hypermutations of immunoglobulin variable regions, that are present in B cells and a variety of non-B cells, appear to be missing in yeast. In contrast to APOBEC-1, AID alone does not exhibit an intrinsic specificity for its target sequences.

Predictors of early relapse in postmenopausal women with hormone receptor-positive breast cancer in the BIG 1-98 trial

BACKGROUND: Aromatase inhibitors are considered standard adjuvant endocrine treatment of postmenopausal women with hormone receptor-positive breast cancer, but it remains uncertain whether aromatase inhibitors should be given upfront or sequentially with tamoxifen. Awaiting results from ongoing randomized trials, we examined prognostic factors of an early relapse among patients in the BIG 1-98 trial to aid in treatment choices. PATIENTS AND METHODS: Analyses included all 7707 eligible patients treated on BIG 1-98. The median follow-up was 2 years, and the primary end point was breast cancer relapse. Cox proportional hazards regression was used to identify prognostic factors. RESULTS: Two hundred and eighty-five patients (3.7%) had an early relapse (3.1% on letrozole, 4.4% on tamoxifen). Predictive factors for early relapse were node positivity (P < 0.001), absence of both receptors being positive (P < 0.001), high tumor grade (P < 0.001), HER-2 overexpression/amplification (P < 0.001), large tumor size (P = 0.001), treatment with tamoxifen (P = 0.002), and vascular invasion (P = 0.02). There were no significant interactions between treatment and the covariates, though letrozole appeared to provide a greater than average reduction in the risk of early relapse in patients with many involved lymph nodes, large tumors, and vascular invasion present. CONCLUSION: Upfront letrozole resulted in significantly fewer early relapses than tamoxifen, even after adjusting for significant prognostic factors.

Abdominal magnetic resonance imaging in small rodents using a clinical 1.5 T MR scanner

Because of superior soft-tissue contrast compared to other imaging techniques, non-invasive abdominal magnetic resonance imaging (MRI) is ideal for monitoring organ regeneration, tissue repair, cancer stage, and treatment effects in a wide variety of experimental animal models. Currently, sophisticated MR protocols, including technically demanding procedures for motion artefact compensation, achieve an MRI resolution limit of < 100 microm under ideal conditions. However, such a high spatial resolution is not required for most experimental rodent studies. This article describes both a detailed imaging protocol for MR data acquisition in a ubiquitously and commercially available 1.5 T MR unit and 3-dimensional volumetry of organs, tissue components, or tumors. Future developments in MR technology will allow in vivo investigation of physiological and pathological processes at the cellular and even the molecular levels. Experimental MRI is crucial for non-invasive monitoring of a broad range of biological processes and will further our general understanding of physiology and disease.

Site-1 protease is essential for endochondral bone formation in mice

Site-1 protease (S1P) has an essential function in the conversion of latent, membrane-bound transcription factors to their free, active form. In mammals, abundant expression of S1P in chondrocytes suggests an involvement in chondrocyte function. To determine the requirement of S1P in cartilage and bone development, we have created cartilage-specific S1P knockout mice (S1P(cko)). S1P(cko) mice exhibit chondrodysplasia and a complete lack of endochondral ossification even though Runx2 expression, Indian hedgehog signaling, and osteoblastogenesis is intact. However, there is a substantial increase in chondrocyte apoptosis in the cartilage of S1P(cko) mice. Extraction of type II collagen is substantially lower from S1P(cko) cartilage. In S1P(cko) mice, the collagen network is disorganized and collagen becomes entrapped in chondrocytes. Ultrastructural analysis reveals that the endoplasmic reticulum (ER) in S1P(cko) chondrocytes is engorged and fragmented in a manner characteristic of severe ER stress. These data suggest that S1P activity is necessary for a specialized ER stress response required by chondrocytes for the genesis of normal cartilage and thus endochondral ossification.