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Background: Transarterial chemoembolization (TACE) is indicated for unresectable hepatocellular carcinoma., Methods: This was a retrospective study of 50 hepatocellular carcinoma patients treated with TACE using doxorubicin-loaded LifePearl™ to investigate the safety and efficacy of TACE., Results: There was no 30-day mortality, and limited adverse events were reported. At best tumor response, complete response and disease control were 58% and 94%, respectively, with a median of 4.5 months of follow-up. Median overall survival was 33.8 months. Patients with Barcelona Clinic Liver Cancer stage 0 and stage A at best tumor response showed a higher complete response rate (83%) than patients with Barcelona Clinic Liver Cancer stage B (complete response: 50%; p = 0.0414)., Conclusion: Doxorubicin-loaded LifePearl™ TACE might be an effective treatment, with a good safety profile, for patients with early/intermediate-stage hepatocellular carcinoma. Further prospective data, especially with a small cohort of selected patients, are required to confirm these results., Competing Interests: Financial & competing interests disclosure This study was funded by a research grant from Terumo Europe NV (Leuven, Belgium). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript., (© 2021 Gerardo Tovar-Felice.)

Current archaeological paradigm proposes that the first peopling of the Americas does not exceed the Last Glacial Maximum period. In this context, the acceptance of the anthropogenic character of the earliest stone artefacts generally rests on the presence of projectile points considered no more as typocentric but as typognomonic, since it allows, by itself, to certify the human character of the other associated artefacts. In other words, without this presence, nothing is certain. Archaeological research at Piauí (Brazil) attests to a Pleistocene human presence between 41 and 14 cal kyr BP, without any record of lithic projectile points. Here, we report the discovery and interpretation of an unusual stone artefact in the Vale da Pedra Furada site, in a context dating back to 24 cal kyr BP. The knapping stigmata and macroscopic use-wear traces reveal a conception centred on the configuration of double bevels and the production in the same specimen of at least two successive artefacts with probably different functions. This piece unambiguously presents an anthropic character and reveals a technical novelty during the Pleistocene occupation of South America., Competing Interests: The authors have declared that no competing interests exist.

The European Union (EU) continuously takes ensuring the safe use of manufactured nanomaterials (MNMs) in consumer products into consideration. The application of a common approach for testing MNMs, including the use of optimized protocols and methods' selection, becomes increasingly important to obtain reliable and comparable results supporting the regulatory framework. In the present study, we tested four representative MNMs, two titanium dioxides (NM100 and NM101) and two silicon dioxides (NM200 and NM203), using the EU FP7-NANoREG approach, starting from suspension and dispersion preparations, through to their characterization and final evaluation of biological effects. MNM dispersions were prepared following a refined NANOGENOTOX protocol and characterized by dynamic light scattering (DLS) in water/bovine serum albumin and in media used for in vitro testing. Potential genotoxic effects were evaluated on human bronchial BEAS-2B cells using micronucleus and Comet assays, and pro-inflammatory effects by cytokines release. Murine macrophages RAW 264.7 were used to detect potential innate immune responses using two functional endpoints (pro-inflammatory cytokines and nitric oxide [NO] production). The interaction of MNMs with RAW 264.7 cells was studied by electron microscopy. No chromosomal damage and slight DNA damage and an oxidative effect, depending on MNMs, were observed in bronchial cells. In murine macrophages, the four MNMs directly induced tumor necrosis factor α or interleukin 6 secretion, although at very low levels; lipopolysaccharide-induced NO production was significantly decreased by the titania and one silica MNM. The application of this approach for the evaluation of MNM biological effects could be useful for both regulators and industries., (© 2020 John Wiley & Sons, Ltd.)

Respiratory allergies are known to affect people all over the world. Environmental factors related to pollution play a significant etiopathogenic role in this regard. Polluting sources are industrial activities and urban traffic, capable of generating various types of pollutants that trigger inflammatory, direct, and indirect damage to tissues, promoting allergic symptoms, even serious ones, and interfering with the pharmacologic response. They are also able to modify pollen, promoting allergic sensitization. Pollution could have played a significant predisposing role in the ongoing morbidity and mortality of SARS-CoV-2., (© 2020 European Academy of Allergy and Clinical Immunology and John Wiley & Sons Ltd.)

Aim: In many cases, the dynamics of psychotherapeutic change processes is characterized by sudden and critical transitions. In theoretical terms, these transitions may be "phase transitions" of self-organizing nonlinear systems. Meanwhile, a variety of methods is available to identify phase transitions even in short time series. However, it is still an open question if different methods for timeseries analysis reveal convergent results indicating the moments of critical transitions and related precursors., Methods and Procedures: Seven concepts which are commonly used in nonlinear time series analysis were investigated in terms of their ability to identify changes in psychological time series: Recurrence Plots, Change Point Analysis, Dynamic Complexity, Permutation Entropy, Time Frequency Distributions, Instantaneous Frequency, and Synchronization Pattern Analysis, i.e., the dynamic inter-correlation of the system's variables. Phase transitions were simulated by shifting control parameters in the Hénon map dynamics, in a simulation model of psychotherapy processes (one by an external shift of the control parameter and one created by a simulated control parameter shift), and three sets of empirical time series generated by daily self-ratings of patients during the treatment., Results: The applied methods showed converging results indicating the moments of dynamic transitions within an acceptable tolerance. The convergence of change points was confirmed statistically by a comparison to random surrogates. In the three simulated dynamics with known phase transitions, these could be identified, and in the empirical cases, the methods converged indicating one and the same transition (possibly the phase transitions of the cases). Moreover, changes that did not manifest in a shift of mean or variance could be detected., Conclusion: Changes can occur in many different ways in the psychotherapeutic process. For instance, there can be very slow and small transitions or very high and sudden ones. The results show the validity and stability of different measures indicating pattern transitions and/or early warning signals of those transitions. This has profound implications for real-time monitoring in psychotherapy, especially in cases where a transition is not obvious to the eye. Reliably identifying points of change is mandatory also for research on precursors, which in turn can help improving treatment., (Copyright © 2020 Schiepek, Schöller, de Felice, Steffensen, Bloch, Fartacek, Aichhorn and Viol.)

Aim: Psychotherapy could be interpreted as a self-organizing process which reveals discontinuous pattern transitions (so-called phase transitions). Whereas this was shown in the conscious process of awake patients by different measures and at different time scales, dreams came very seldom into the focus of investigation. The present work tests the hypothesis that, by dreaming, the patient gets progressively more access to affective-laden (i.e., emotionally charged) unconscious dimensions. Furthermore, the study investigates if, over the course of psychotherapy, a discontinuous phase transition occurs in the patient's capacity to get in contact with those unconscious dimensions., Methods and Procedures: A series of 95 dream narratives reported during a psychoanalytic psychotherapy of a female patient (published as the "dreams of Amalie X") was used for analysis. An automated text analysis procedure based on multiple correspondence analysis was applied to the textual corpus of the dreams, highlighting a 10-factor structure. The factors, interpreted as affective-laden unconscious meaning dimensions, were adopted to define a 10-dimensional phase space, in which the ability of a dream to be associated with one or more local factors representing complex affective-laden meanings is measured by the Euclidean distance (ED) from the origin of this hyperspace. The obtained ED time series has been fitted by an autoregressive integrated moving average (ARIMA) model and by non linear methods like dynamic complexity, recurrence plot, and time frequency distribution. Change point analysis was applied to these non linear methods., Results: The results show an increased frequency and intensity of dreams to get access to affective-laden meanings. Non linear methods identified a phase transition-like jump of the ED dynamics onto a higher complexity level of the dreaming process, suggesting a non linear process in the patient's capacity to get in contact with unconscious dimensions., Conclusion: The study corroborates the hypothesis that, by dreaming, the patient gets progressively more access to affective-laden meaning intended as unconscious dimensions. The trajectory of this process has been reproduced by an ARIMA model, and beyond this, non linear methods of time series analysis allowed the identification of a phase transition in the unconscious process of the psychoanalytic therapy under investigation., (Copyright © 2020 Gennaro, Kipp, Viol, de Felice, Andreassi, Aichhorn, Salvatore and Schiepek.)

Statistical mechanics is the field of physics focusing on the prediction of the behavior of a given system by means of statistical properties of ensembles of its microscopic elements. The authors examined the possibility of applying such an approach to psychotherapy research with the aim of investigating (a) the possibility of predicting good and poor outcomes of psychotherapy on the sole basis of the correlation pattern among their descriptors and (b) the analogies and differences between the processes of good- and poor-outcome cases. This work extends the results reported in a previous paper and is based on higher-order statistics stemming from a complex network approach. Four good-outcome and four poor-outcome brief psychotherapies were recorded, and transcripts of the sessions were coded according to Mergenthaler's Therapeutic Cycle Model (TCM), i.e., in terms of abstract language, positive emotional language, and negative emotional language. The relative frequencies of the three vocabularies in each word-block of 150 words were investigated and compared in order to understand similarities and peculiarities between poor-outcome and good-outcome cases. Network analyses were performed by means of a cluster analysis over the sequence of TCM categories. The network analyses revealed that the linguistic patterns of the four good-outcome and four poor-outcome cases were grounded on a very similar dynamic process substantially dependent on the relative frequency of the states in which the transition started and ended ("random-walk-like behavior", adjusted R 2 = 0.729, p < 0.001). Furthermore, the psychotherapy processes revealed statistically significant changes in the relative occurrence of visited states between the beginning and the end of therapy, thus pointing to the non-stationarity of the analyzed processes. The present study showed not only how to quantitatively describe psychotherapy as a network, but also found out the main principles on which its evolution is based. The mind, from a linguistic perspective, seems to work-through psychotherapy sessions by passing from the most adjacent states and the most occurring ones. This finding can represent a fertile ground to rethink pivotal clinical concepts such as the timing of an interpretation or a comment, the clinical issue to address within a given session, and the general task of a psychotherapist: from someone who delivers a given technique toward a consultant promoting the flexibility of the clinical field and, thus, of the patient's mind., (Copyright © 2020 de Felice, Giuliani, Gelo, Mergenthaler, De Smet, Meganck, Paoloni, Andreassi, Schiepek, Scozzari and Orsucci.)

Food additive E551 consists of synthetic amorphous silica (SAS), comprising agglomerates and aggregates of primary particles in the nanorange (<100 nm), which potential nanospecific risks for humans associated to dietary exposure are not yet completely assessed. In NANoREG project, aim of the study was to identify potential hazards of pyrogenic SAS nanomaterial NM-203 by a 90-day oral toxicity study (OECD test guideline 408). Adult Sprague-Dawley rats of both sexes were orally treated with 0, 2, 5, 10, 20 and 50 mg SAS/kg bw per day; dose levels were selected to be as close as possible to E551 dietary exposure. Several endpoints were investigated, the whole integrative study is presented here along with the results of dispersion characterization, tissue distribution, general toxicity, blood/serum biomarkers, histopathological and immunotoxicity endpoints. No mortality, general toxicity and limited deposition in target tissues were observed. NM-203 affected liver and spleen in both sexes. Proposed NOAEL 5 mg/kg bw per day in male rats for enlarged sinusoids in liver. In female rats, TSH and creatinine levels were affected, proposed LOAEL 2 mg/kg bw per day. Overall, these data provide new insight for a comprehensive risk assessment of SAS exposure by the oral route., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: Childhood maltreatment (CM) is recognized as a non-specific risk factor for Eating Disorders (EDs), but the mechanisms explaining this association have been insufficiently assessed. We aim to explore the psychological pathways through which CM experiences promote ED core symptoms., Methods: Two-hundred-twenty-eight people with EDs, 94 with anorexia nervosa restricting (ANR) type and 134 with binge-purging (BP) symptoms (including 23 with AN purging type and 111 with bulimia nervosa), completed the Eating Disorder Inventory-2, the State-Trait Anxiety Inventory and the Childhood Trauma Questionnaire. The variables provided by these questionnaires were included in a network analysis to identify the shortest pathways between CM nodes and ED core symptoms. Then mediation analysis was performed in order to confirm the mediation role of the nodes included in the shortest pathways from CM to ED core symptoms., Results: All types of CM experiences were connected to the ED psychopathology through emotional abuse. In the ANR group, interoceptive awareness was included in the shortest path between emotional abuse and drive to thinness and mediated this relationship. In the BP group, the shortest routes between CM and ED core symptoms included both ineffectiveness and interoceptive awareness., Conclusions: Combining the network analysis approach with the mediation analyses provides for the first time a putative hybrid model, which reveals that all CM types converge towards ED symptoms through emotional abuse and that interoceptive awareness and ineffectiveness mediate these connections in people with ANR and BP symptoms, respectively. These findings may have possible implications for both research and treatment of EDs., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

An early dialogue between nanomedicine developers and regulatory authorities are of utmost importance to anticipate quality and safety requirements for these innovative health products. In order to stimulate interactions between the various communities involved in a translation of nanomedicines to clinical applications, the European Commission's Joint Research Centre hosted a workshop titled "Bridging communities in the field of Nanomedicine" in Ispra/Italy on the 27th -28th September 2017. Experts from regulatory bodies, research institutions and industry came together to discuss the next generation of nanomedicines and their needs to obtain regulatory approval. The workshop participants came up with recommendations highlighting methodological gaps that should be addressed in ongoing projects addressing the regulatory science of nanomedicines. In addition, individual opinions of experts relevant to progress of the regulatory science in the field of nanomedicine were summarised in the format of a survey., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Polybrominated diphenyl ethers (PBDEs) are persistent organic pollutants that are added to numerous products to prevent accidental fires. PBDEs are present in the environment and they bio-accumulate in human and animal tissues. Recently, their presence has been correlated to several pathologies but little is known about their effect on the human innate immune system activity. In this study we investigated the effect of the congener 2,2',4,4'-Tetrabromodiphenyl ether (PBDE-47) on the functional activity of the THP-1 human macrophages cell line and on ex vivo freshly isolated human basophils. Cytotoxicity and genotoxicity studies showed that PBDE-47 was able to induce toxic effects on the THP-1 cell line viability at concentrations ≥25 μM. Immune function of THP-1 was studied after stimulation with bacterial lipopolysaccharide (LPS) and PBDE-47 exposure at concentrations granting macrophage viability. Two dimensional electrophoresis showed modification of the proteome in the 3 μM PBDE-47 treated sample and Real Time PCR and ELISA demonstrated a statistically significant reduction in the expression of IL-1β, IL-6 and TNF-α cytokines. Furthermore, PBDE-47 was able to perturbate genes involved in cell motility upregulating CDH-1 and downregulating MMP-12 expressions. Finally, basophil activation assay showed reduced CD63 activation in PBDE-47 treated samples. In conclusion, our study demonstrated that PBDE-47 may perturb the activities of cells involved in innate immunity dampening the expression of macrophage pro-inflammatory cytokines (IL-1β, IL-6 and TNF-α) and genes involved in cell motility (MMP-12 and E-cadherin) and interfering with basophil activation suggesting that this compound can impair innate immune response., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

A systematic single case study with a mixed quantitative and qualitative methodology was conducted to investigate the nonlinear dynamics of change in play profiles of a child in psychodynamic play therapy. The first aim of the study was to cluster the different features of play characteristics (i.e., descriptive, cognitive, affective, and social characteristics of play, and the defensive strategies used in play) in order to construct the different play profiles of this child, and secondly to assess the transitions between profiles over the course of treatment. It was expected that there would be an increase in critical fluctuations during the transitions between the profiles and an increase in the variability of play profiles. Results showed that the child's play characteristics clustered into eight states and three attractors. The Markov Transition Matrix showed how play profiles evolved over time. Entropy analyses comparing the first and the second half of therapy showed an increase in variability. Qualitative analyses indicated the importance of expression of the child's underlying fear, and its integration with overt anger in the generation of the new play profiles. The results indicate an increase in variability, and a destabilization of old play profiles that were used towards generating new play profiles.

Purpose: To describe the prevalent clinical, laboratory, and radiological features of otogenic lateral sinus thrombosis (OLST) in children; to identify clinical predictors of outcome; to propose a management algorithm derived from experience., Methods: A retrospective review was conducted of the clinical records of patients with OLST, treated in a single tertiary care referral center for pediatric disease from 2006 to 2017. The inclusion criteria were pediatric age (0-16 years) and OLST diagnosis confirmed by a pre- and post-contrast CT or venography-MRI scan. Primary outcome measures were early (1-2 months) and late (6 months) sinus recanalization assessed by means of neuroimaging., Results: Twenty-five patients (8 females and 17 males; mean age = 6 ± 3 years) were included. A genetic abnormality associated with thrombophilia was found in 24 (96%) patients. At diagnosis, anticoagulant treatment with low-molecular-weight heparin (LMWH) was started in all subjects, while surgical treatment (mastoidectomy and tympanostomy tube insertion) was performed in 16/25 (64%) patients. Follow-up neuroimaging showed lateral sinus recanalization in 12/25 (48%) patients after 1-2 months and in 17/25 (68%) after 6 months. At multivariate logistic regression analysis, no significant predictors of the early and late neuroradiological outcome were found., Conclusions: All children with OLST should be screened for thrombophilia to decide on treatment duration and to assess the need for future antithrombotic prophylaxis. Immediately after diagnosis, anticoagulant treatment with LMWH should be started according to the international guidelines. Instead, our experience suggests that surgical treatment should not be indicated in all patients, but decided on a case-to-case basis.

Study Objective: To assess the association between ovarian endometriomas detectable at transvaginal ultrasound (TVS) and other specific extraovarian lesions including adhesions, deep infiltrating endometriosis (DIE), and adenomyosis., Design: Retrospective observational study (Canadian Task Force classification II-2)., Setting: Two university hospitals., Patients: Two hundred fifty-five symptomatic women with at least 1 ovarian endometrioma found on ultrasound after presentation with pain or irregular menstruation., Interventions: Patients underwent TVS followed by either medical or surgical treatment., Measurements and Main Results: Two hundred fifty-five women, aged 20 to 40 years, underwent TVS and were found to have at least 1 endometrioma with a diameter > 20 mm. Associated sonographic signs of pelvic endometriosis (adhesions, DIE, and adenomyosis) were recorded, and a subgroup of patients (n = 50) underwent laparoscopic surgery within 3 months of TVS. Mean endometrioma diameter was 40.0 ± 18.1 mm, and bilateral endometriomas were observed in 65 patients (25.5%). TVS showed posterior rectal DIE in 55 patients (21.5%) and a thickening of at least 1 uterosacral ligament in 93 patients (36.4%). One hundred eighty-six patients (73%) had adhesions, and 134 patients (53%) showed signs of myometrial adenomyosis on TVS. Thirty-eight patients (15%) exhibited only a single isolated endometrioma with a mobile ovary and no other signs of pelvic endometriosis/adenomyosis at TVS., Conclusion: Ovarian endometriomas are indicators for pelvic endometriosis and are rarely isolated. Particularly, left endometriomas were found to be associated with rectal DIE and left uterosacral ligament localization and bilateral endometriomas correlated with adhesions and pouch of Douglas obliteration, whereas no correlation was found between endometrioma size and DIE. Determining appropriate management, whether clinical or surgical, is critical for ovarian endometriomas and concomitant adhesions, endometriosis, and adenomyosis in patients desiring future fertility., (Copyright © 2018 American Association of Gynecologic Laparoscopists. Published by Elsevier Inc. All rights reserved.)

Introduction: Impaired thrombin generation has been associated to increase bleeding after cardiac surgery with cardiopulmonary bypass (CPB), especially in children. The aim of this study was to evaluate standard coagulation assay, thrombin generation by calibrated automated thrombogram (CAT), thromboelastography (TEG) and procoagulant phospholipids (PPL) activity in infants undergoing cardiac surgery with CPB., Materials and Methods: Prospective observational study performed in children aged <24months undergoing cardiac surgery with CPB. Exclusion criteria were preoperative coagulopathy or anticoagulant therapy. Coagulation was evaluated by standard coagulation assays (prothrombin time, activated partial thromboplastin time, fibrinogen level, platelet count), TEG, CAT and PPL at anaesthesia induction (T1) and after 12h (T2). Perioperative bleeding management was performed according to the institutional guidelines., Results: Forty-nine children aged <24months were enrolled. At T1 ETP and peak height evaluated by CAT were significantly lower in infants aged <6months. Standard coagulation tests, TEG and PPL did not correlate with age. At T2 platelet count, plasmatic fibrinogen level, all TEG parameters, ETP and peak height by CAT were significantly impaired compared to baseline values (T1), despite allogeneic blood product transfusions., Conclusions: Thrombin generation is significantly impaired in children affected by congenital heart disease, compared to healthy children and adults. CAT parameters resulted age-dependent, and thrombin generation is lower in infants aged <6months. After cardiac surgery with CPB, a coaugulopathy, revealed by CAT, TEG, but not by PT and aPTT assays, is persistent 12h after surgery despite transfusions of blood products., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Bones of human and ground sloth remains were analyzed for presence of Trypanosoma cruzi by conventional PCR using primers TC, TC1 and TC2. Sequence results amplified a fragment with the same product size as the primers (300 and 350pb). Amplified PCR product was sequenced and analyzed on GenBank, using Blast. Although these sequences did not match with these parasites they showed high amplification with species of bacteria. This article presents the methodology used and the alignment of the sequences. The display of this dataset will allow further analysis of our results and discussion presented in the manuscript "Finding the unexpected: a critical view on molecular diagnosis of infectious diseases in archaeological samples" (Pucu et al. 2017) [1].

Background: Anisakis simplex is the only fishery-product associated parasite causing clinical allergic responses in humans so far. However, other anisakids, due to the presence of shared or own allergens, could also lead to allergic reactions after sensitization. The aim of this study was to determine if Pseudoterranova decipiens belonging to the family Anisakidae has allergenic activity and is able to induce sensitization after oral administration in a murine (BALB/c mice) model., Results: The ingestion of A. pegreffii proteins by BALB/c mice, which had been previously sensitized by intraperitoneal inoculation with the corresponding live L3 larvae, triggers signs of allergy within 60 min, whereas P. decipiens did to a lesser extent. Beside symptoms, allergic reactions were furtherly supported by the presence of histamine in sera of sensitized mice. Specific IgG1 and IgE responses were detected in sera of all sensitized mice from week four. Specific IgG2a response was detected in sera from mice sensitized to P. decipiens. After polyclonal or specific activation with anti-CD3/anti-CD28 or antigens, respectively, splenocytes from mice infected i.p. with A. pegreffii or P. decipiens larvae showed significantly higher production of IL-10 than naïve mice. After stimulation with specific antigens, significantly higher IL-5 and IL-13 amounts were produced by specific antigen stimulated splenocytes than by the naïve cells; only P. decipiens proteins induced IFN-ɣ., Conclusions: The overall results suggest that infection with P. decipiens can sensitize mice to react to subsequent oral challenge with anisakid proteins, as described for A. simplex (sensu stricto) and A. pegreffii infections. The results show that anisakid proteins induce a dominant Th2 response, although P. decipiens could also induce a mixed type 1/type 2 pattern.

Background and Aims: Several studies report the use of thromboelatography (TEG) to monitor coagulation in pediatric cardiac surgery. The aim of this study was to compare baseline and intraoperative TEG, TEG-functional fibrinogen, and standard coagulation assays in children with cyanotic and acyanotic congenital heart disease (CHD) undergoing cardiac surgery., Methods: This is a prospective observational study of 63 children aged <24 months undergoing cardiac surgery with cardiopulmonary bypass (CPB). Exclusion criteria included preoperative anticoagulant therapy and hepatic failure. We collected blood at anesthesia induction (T1), at lowest temperature after CPB start (T2), and after heparin neutralization (T3). Coagulation was evaluated by TEG (reaction time [R]), k, alpha-angle, maximum amplitude (MA), MA-fibrinogen (MA-fib), and by standard coagulation assays (prothrombin time, activated partial thromboplastin time, fibrinogen level, platelet [PLT] count)., Results: Sixty-three patients were enrolled (38 cyanotic and 25 acyanotic). Median age was 4 [IQR 2-6] months and median weight was 5 [IQR 3.7-6.5] kg. Most common surgeries were: ventricular septal defect repair (n = 13), Fallot correction (n = 11), and arterial switch operation (n = 10). Cyanotic and acyanotic children were well matched: R, k, MA, and MA-fib at T1, T2, and T3 were not significantly different between cyanotic and acyanotic children. At T2, significant correlations were showed between MA and PLT count (r = 0.4; P = 0.0008) and k and plasma fibrinogen level (r = -0.54; P < 0.0001). At T3, significant correlations were showed between MA and PLT count (r = 0.5; P < 0.0001), G and PLT count (r = 0.6; P < 0.0001), and MA-fib and plasma fibrinogen level (r = 0.5; P = 0.002)., Conclusions: According to our findings, cyanosis does not affect TEG parameters in children with CHD. PLT count and plasma fibrinogen significantly correlated (are significantly associated) with MA and MA-fib respectively, suggesting that use of TEG after protamine administration may be prompted for improved hemostatic monitoring in the perioperative phase., (© 2016 John Wiley & Sons Ltd.)

Aim: Even though there is substantial evidence that play based therapies produce significant change, the specific play processes in treatment remain unexamined. For that purpose, processes of change in long-term psychodynamic play therapy are assessed through a repeated systematic assessment of three children's "play profiles," which reflect patterns of organization among play variables that contribute to play activity in therapy, indicative of the children's coping strategies, and an expression of their internal world. The main aims of the study are to investigate the kinds of play profiles expressed in treatment, and to test whether there is emergence of new and more adaptive play profiles using dynamic systems theory as a methodological framework. Methods and Procedures: Each session from the long-term psychodynamic treatment (mean number of sessions = 55) of three 6-year-old good outcome cases presenting with Separation Anxiety were recorded, transcribed and coded using items from the Children's Play Therapy Instrument (CPTI), created to assess the play activity of children in psychotherapy, generating discrete and measurable units of play activity arranged along a continuum of four play profiles: "Adaptive," "Inhibited," "Impulsive," and "Disorganized." The play profiles were clustered through K -means Algorithm, generating seven discrete states characterizing the course of treatment and the transitions between these states were analyzed by Markov Transition Matrix, Recurrence Quantification Analysis (RQA) and odds ratios comparing the first and second halves of psychotherapy. Results: The Markov Transitions between the states scaled almost perfectly and also showed the ergodicity of the system, meaning that the child can reach any state or shift to another one in play. The RQA and odds ratios showed two trends of change, first concerning the decrease in the use of "less adaptive" strategies, second regarding the reduction of play interruptions. Conclusion: The results support that these children express different psychic states in play, which can be captured through the lens of play profiles, and begin to modify less dysfunctional profiles over the course of treatment. The methodology employed showed the productivity of treating psychodynamic play therapy as a complex system, taking advantage of non-linear methods to study psychotherapeutic play activity.

Objective: To study outcomes and complications during pregnancy and at delivery in women with a posterior deep infiltrating endometriosis (DIE) nodule persisting after surgery and diagnosed at transvaginal sonography (TVS) in comparison with a control group of women without endometriosis., Design: Multicenter observational and cohort study., Setting: University hospital., Patient(s): Women (n = 200) with a posterior DIE nodule equal or more than 2 cm centimeters in size who desired a pregnancy and a control group of women (n = 300) with no previous recorded diagnosis of endometriosis who delivered in our clinic during the same time period., Intervention(s): Patient data collected from medical charts and by phone interviews., Main Outcome Measure(s): Evaluation of complications during pregnancy and delivery., Result(s): Of the 101 women with a posterior DIE nodule, 52 become pregnant among whom 25 used assisted reproductive technology. Of these 52 pregnancies, 11 ended in an early abortion, and 41 delivered a baby; 13 (31.7%) had a premature delivery, 7 (17.8%) a placenta praevia, and 28 (68.2%) had a cesarean delivery. When compared with the control group, the women with endometriosis had a higher risk of pregnancy complicated by preterm birth, placenta previa, placental abruption, and hypertension. Cesarean delivery and complications during surgery (hysterectomy, hemoperitoneum, and bladder injuries) were statistically significantly more frequent in women with endometriosis than in controls., Conclusion(s): Women with an incomplete removal of posterior DIE have a high complications rate during pregnancy and delivery., (Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

Purpose: The development, analysis, and first clinical use of a novel eye drop formulation of plasminogen and hyaluronate sodium for the treatment of a patient with ligneous conjunctivitis (LC) are described., Summary: LC is a chronic inflammatory disorder of the eye that can in rare cases lead to corneal involvement and subsequent blindness. Topically administered plasminogen is an effective treatment for LC; however, the lack of a specific and validated formulation of plasminogen for topical treatment can constitute a gap in the quality of care. A novel formulation of plasminogen and hyaluronate sodium for the treatment of LC was developed by combining commercially available products. Through a series of tests, including microbiological, viscosity, and pH analyses, the novel eye drop formulation was demonstrated to have constant activity (3.3 IU/mL or greater), to be microbiologically stable (i.e., sterile), and to be safe enough for daily administration. The first clinical application of the novel eye drop formulation was in the case of a nine-year-old girl with LC affecting both eyes. Initially, the girl's parents administered two drops every three hours to the left eye each day while the girl was awake. On examination after 30 days of daily use of the eye drop formulation, the patient was found to have a clear cornea and no membrane development on the conjunctiva; the formulation was subsequently administered in both eyes, with positive results., Conclusion: A new formulation of plasminogen plus hyaluronate sodium was developed, tested, and used successfully in a girl with LC., (Copyright © 2016 by the American Society of Health-System Pharmacists, Inc. All rights reserved.)

We studied the synchronization dynamics of a therapist and patient during a psychotherapy session. This investigation was developed in order to explore a new possible perspective and methodology for studying the expression of emotions. More specifically, literature concerning synchronization of in-session non-verbal variables emphasises its positive correlation with empathy and therapeutic outcomes. We compared the dynamics of galvanic skin response (GSR) and linguistic prosody, chosen as indicators of emotional expression in different domains. We studied their synchronization through complementary methodologies: Recurrence Quantification Analysis (RQA) and Principal Component Analysis (PCA), Markov Transition Matrix (MTM) and Cross-Recurrence Quantification Analysis (CRQA). We investigated the nonlinearity of GSR in terms of self-similarity and power-law, as emerged in autocorrelation functions and signal variations. We considered time-lagged correlations as a measure of dynamical systems' memory. This article concludes by highlighting the importance of a deeper study of all variables related to the psychotherapeutic process and their synchronization in order to extend our knowledge of general human dynamics.

Allergen specific immunotherapy has been introduced in the clinic more than 100 years ago showing effectiveness and, so far, it represents the only curative approach to treat allergic disorders ameliorating the symptoms, reducing the medication costs and blocking the onset of new sensitizations. However, some questions are still open regarding to the safety of the treatment and the need to reduce the dose and time of administration to improve the compliance of the patients. All preparations that are currently available may trigger side effects. For these reasons, new formulations and route of administration have been exploited demonstrating that such products presented improved efficacy and safety. Nanotechnology for biomedical applications offers many advantages, such as improved stability and bioavailability, favourable biodistribution profiles and targeting to specific cell populations whose impact on the immune system has been evaluated in animal systems. Nanoparticles interact with the immune system, and the final outcome of this interaction depends on their physico-chemical characteristics. Concerns can be raised when immunotoxic effect are induced, resulting in inflammatory dangerous responses or in the reduction of the normal defensive activity of the immune system. In this paper, we will review the most relevant data about the synthesis of allergen/nanoparticles systems and will discuss their impact on the immune system in terms of immunomodulatory activity and immunotoxicity risk assessment.

In previous studies it has been demonstrated that the plant alkaloid voacamine (1), used at noncytotoxic concentrations, enhanced the cytotoxicity of doxorubicin and exerted a chemosensitizing effect on cultured multidrug-resistant (MDR) U-2 OS-DX osteosarcoma cells. The in vitro investigations reported herein gave the following results: (i) the chemosensitizing effect of 1, in terms of drug accumulation and cell survival, was confirmed using SAOS-2-DX cells, another MDR osteosarcoma cell line; (ii) compound 1 enhanced the cytotoxic effect of doxorubicin also on the melanoma cell line Me30966, intrinsically drug resistant and P-glycoprotein-negative; (iii) at the concentrations used to sensitize tumor cells, 1 was not cytotoxic to normal cells (human fibroblasts). These findings suggest possible applications of voacamine (1) in integrative oncologic therapies against resistant tumors.

The cone-rod homeobox (CRX) is a gene that belongs to the member of the orthodenticle homeobox (Otx) family, with important function in development and differentiation of retinal and pineal cells. Moreover, CRX appears to be specifically expressed in pineal tumors and retinoblastomas. We performed an immunohistochemical study on 91 pediatric and adult central nervous system tumors, plus 2 normal brain samples. Our results demonstrated that CRX is expressed not only in pineal parenchymal tumors and retinoblastoma, but also in a some medulloblastomas and supratentorial primitive neuroectodermal tumors. None of the glial tumors screened were positive for CRX. In conclusion, CRX could be useful in surgical neuropathology for the differential diagnosis of pineal region tumors, in particular to discriminate pineal tumors from glial tumors.

Objective: To evaluate the characteristics of the uterine junctional zone (JZ) by three-dimensional (3D) transvaginal sonography (TVS) in women with recurrent miscarriage (RM) as compared to normal fertile controls., Study Design: The thickness and the morphology of the JZ were evaluated in 75 women with a history of RM due to different causes and in 20 fertile women without a history of miscarriages or pelvic disease. All patients included in the study were selected among those who attended the outpatient clinic of "Tor Vergata" University. The JZ characteristics were evaluated in the midluteal phase of the cycle on the uterine coronal section obtained by 3D TVS., Results: Patients with RM showed a JZ maximum thickness significantly increased when compared to that observed in control group (5.8±0.7 vs. 5.0±1.1mm). When grouped according to the different causes of RM, all groups of patients with RM showed an increased JZ thickness when compared to fertile women, with the exception of those with anti-phospholipid antibody syndrome, probably due to the small number of cases with this pathology., Conclusions: A thickened JZ could be an independent indicator of the risk of miscarriage and may represent an important contributing factor to some causes of RM. These observations may offer new perspectives for the screening and treatment of patients with RM. Although further studies are needed to ascertain if the reduction of the JZ thickness can determine a better pregnancy outcome, 3D TVS evaluation of the JZ could provide the opportunity to identify women in which appropriate therapeutic protocols can improve the possibility of successful pregnancy., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Scope: Among food allergies, peanut allergy is frequently associated with severe anaphylactic reactions. In the need for safe and effective therapeutic strategies, probiotics may be considered on the basis of their immunomodulatory properties. The aim of the present study was to investigate the immunological mediators involved in the effects of probiotic VSL#3 oral supplementation on Th2 inflammation and anaphylaxis in a mouse model of peanut allergy., Methods and Results: VSL#3 supplementation to peanut-sensitized mice was effective in ameliorating anaphylaxis and Th2-mediated inflammation, by promoting regulatory responses in the jejunum mucosa and in the mesenteric lymph node, as evaluated by ELISA, real-time PCR, histologic, and immunohistochemical analysis. Probiotic-induced TGF-β mediates its protective effects through the induction of regulatory T cells expressing FOXP3 and/or latency-associated peptide, as proven by in vivo blockade of TGF-β in VSL#3-treated mice with a neutralizing monoclonal antibody one day before challenge., Conclusion: TGF-β, induced in the gut by VSL#3 supplementation, is capable of reducing the Th2 inflammation associated with food anaphylaxis in a mouse model of peanut sensitization. TGF-β acts through the induction/maintenance of regulatory T cells expressing FOXP3 and/or latency-associated peptide. Probiotics supplementation may represent an effective and safe strategy for treating food allergies in adult population., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Objective: The uterine junctional zone (JZ) alterations are correlated with adenomyosis. An accurate evaluation of the JZ may be obtained by 3-dimensional transvaginal sonography (TVS). The aim of the present prospective study was to assess the value of detectable alterations by 3-dimensional TVS of the JZ in patients with pelvic endometriosis (diagnosed by laparoscopy and histologic condition) and to compare these findings with those of women without pelvic endometriosis., Study Design: Eighty-two patients who were scheduled for laparoscopy had undergone previous surgery and 2- and 3-dimensional TVS. Uterine multiplanar sections that were obtained by 3-dimensional TVS were used to evaluate JZ features. During laparoscopy, an accurate staging of pelvic endometriosis was performed. JZ thickness and JZ alterations were correlated with stage of endometriosis., Results: Of the 82 patients, 59 patients had endometriosis at laparoscopy and histology. The maximum thickness of JZ in patients with endometriosis was significantly greater than in patients without endometriosis (6.5 ± 1.9 mm vs 4.8 ± 1.0 mm; P < .001). The features of JZ appeared similar at different stages, whereas they are statistically different if correlated with patients without endometriosis., Conclusion: JZ thickness and its alterations are different in patients with endometriosis compared with those women without endometriosis and are not correlated with American Society of Reproductive Medicine staging methods. Because these JZ ultrasound features are associated mostly with adenomyosis, a correlation between endometriosis and JZ hyperplasia and adenomyosis could be hypothesized. Noninvasive evaluation of the JZ may be useful in the identification of those women who are affected by endometriosis also in early stage of the disease when there are no other sonographic signs of pelvic endometriosis., (Copyright © 2013 Mosby, Inc. All rights reserved.)

Background: Shellfish allergy is one of the major causes of life-threatening allergic reactions to food. The shrimp species Pandalus borealis is the commercially most important coldwater shrimp species, and its protein extract is commonly used in shrimp allergy diagnostics. However, the DNA sequence of its major allergen, tropomyosin, designated Pan b 1, was not previously described. Our aim was to identify the cDNA sequence of Pan b 1 and to generate a recombinant protein with similar structure and allergenicity as the natural protein., Methods: P. borealis shrimps were caught in the Oslofjord (Norway). cDNA from Pan b 1 was generated, an N-terminal histidine tag was added, and the protein was expressed in Escherichia coli. The recombinant Pan b 1 was characterized by structural and IgE-binding studies and investigated further with basophil activation tests (BATs) and skin prick tests (SPTs) on Norwegian shrimp-allergic individuals., Results: The open reading frame encoded 284 amino acids that shared 97-100% identity with other shrimp tropomyosins. Mass spectroscopy of natural Pan b 1 confirmed the protein's molecular mass and indicated the absence of posttranslational modifications. Circular dichroism spectroscopy revealed virtually identical spectra between recombinant and natural Pan b 1, which together with native PAGE and size exclusion chromatography results indicated a similar structure. Furthermore, immunoblot and ELISA studies as well as BATs and SPTs showed equivalent results of recombinant and natural Pan b 1., Conclusion: A recombinant tropomyosin from P. borealis was generated that can be used in diagnostics and further studies on tropomyosin allergenicity and specific immunotherapy., (Copyright © 2012 S. Karger AG, Basel.)

It has been twenty years since the first paper reporting the association between thyroid antibodies (TAIs) and spontaneous miscarriage was published. Following this observation, several studies have clearly demonstrated an increased prevalence of TAI in patients with recurrent miscarriage (RM). However, the exact mechanism underlying this association remains a matter of debate. The aim of the present study was to evaluate the thyroid function, throughout a specific test, in patient with RM and TAI focusing on the hypothesis that TAI should be an indirect sign of a mild thyroid dysfunction. 46 patients with RM and TAI were included in the study. All patients underwent short TRH stimulation test showing an abnormal response in the vast majority of cases (65%). Normal FT4 and FT3 mean values were found whereas TSH values were in the upper normal range (2.64 ± 1.3 mUI/L). Our data support the hypothesis that in patients with RM the presence of TAI is an indirect sign of a subtle thyroid dysfunction detectable by a specific test. This test give the possibility to identify women with RM in which specific therapeutic approaches could effectively improve the possibility for a successful pregnancy.

Background: Histopathological grading of ependymoma has been controversial with respect to its reproducibility and clinical significance. In a 3-phase study, we reviewed the pathology of 229 intracranial ependymomas from European trial cohorts of infants (2 trials - SFOP/CNS9204) and older children (2 trials - AIEOP/CNS9904) to assess both diagnostic concordance among five neuropathologists and the prognostic utility of histopathological variables, particularly tumor grading., Results: In phase 1, using WHO criteria and without first discussing any issue related to grading ependymomas, pathologists assessed and independently graded ependymomas from 3 of 4 trial cohorts. Diagnosis of grade II ependymoma was less frequent than grade III, a difference that increased when one cohort (CNS9204) was reassessed in phase 2, during which the pathologists discussed ependymoma grading, jointly reviewed all CNS9204 tumors, and defined a novel grading system based on the WHO classification. In phase 3, repeat independent review of two cohorts (SFOP/CNS9904) using the novel system was associated with a substantial increase in concordance on grading. Extent of tumor resection was significantly associated with progression-free survival (PFS) in SFOP and AIEOP, but not in CNS9204 and CNS9904. Strength of consensus on grade was significantly associated with PFS in only one trial cohort (AIEOP). Consensus on the scoring of individual histopathological features (necrosis, angiogenesis, cell density, and mitotic activity) correlated with PFS in AIEOP, but in no other trial., Conclusions: We conclude that concordance on grading ependymomas can be improved by using a more prescribed scheme based on the WHO classification. Unfortunately, this appears to have utility in limited clinical settings.

The aim of the study was to assess the symptoms prevalence of allergic diseases in a population of 11-15 yr old schoolchildren, to evaluate the associations between asthma and other symptoms and identify risk factors for asthma, rhinitis and eczema syndromes. A sample of 481 students was studied using an International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire. Prevalence of different kind of self-reported symptoms was calculated. Using a logistic regression approach, we tried to identify risk factors for three syndromes - rhinitis, eczema and asthma. The highest and the lowest prevalence rates of self-reported symptoms were recorded for rhinitis (43.6%) and for eczema (8.1%), respectively. The prevalence of asthma was 15.7%. Univariate analysis showed a mutual association between wheeze and rhinitis symptoms. Multivariate logistic regression model for eczema syndrome revealed female gender as a significant risk factor. The polytomic logistic multivariate regression revealed female gender and family history of allergy as significant risk factors for rhinitis syndrome only, and maternal smoking and familial allergy for rhinitis and asthma together. In particular, familial allergy yields a 400% higher chance of developing asthma and rhinitis together. The synergistic effect of familial allergy on rhinitis and asthma syndromes suggests the implementation of preventive measures in children with family history of these diseases.

Background: The immunological mechanisms responsible for the immunomodulatory and anti-allergic effects of probiotic bacteria are still poorly defined. The combined effects of mixtures of different species of probiotic bacteria have been explored only in part. The present study describes the immunomodulatory activity of the VSL#3 probiotic preparation in in vitro and in vivo systems., Methods: The activation and cytokine production by in vitro probiotic-stimulated bone-marrow dendritic cells (BM-DCs) and spleen cells isolated from naïve or Par j 1-sensitized mice were investigated. Mice were intranasally administered a sonicate preparation of VSL#3 before immunization with rPar j 1. Serum antibody levels and cytokine expression in the lung were determined., Results: Both live and sonicated VSL#3 preparations induced maturation and cytokine production by BM-DCs. Cytokine production by spleen cells from naïve or Par j 1-sensitized mice was modulated by the probiotic preparations towards a Treg/Th0 profile, characterized by increased IL-10 and IFN-gamma production. In vivo prophylactic treatment with VSL#3 induced a significant reduction of serum specific IgG1. At lung level, VSL#3 pre-treatment remarkably reduced IL-13 and IL-4 mRNA expression and increased IL-10 expression., Conclusions: The VSL#3 preparations have not only the capacity to bias primary immune responses towards a Treg/Th0-type profile, but also to modify in the same way the functional characteristics of established in vitro Th2 responses. In vivo studies on a mouse model of Par j 1 sensitization indicate that the prophylactic intranasal treatment with probiotic bacteria is able to modulate the development of Th2-biased responses., ((c) 2009 S. Karger AG, Basel.)

Exposure to indoor allergens is an important risk factor for sensitisation and respiratory allergy. The aim of this paper was to evaluate the levels of mite, cat and latex allergens in dust collected from an indoor workplace and to assess whether the exposure to these allergens was associated with the allergy symptoms reported by employees. Sixty dust samples were collected. Allergen concentrations were measured with antibody based ELISAs. All 144 participants compiled a questionnaire exploring possible symptoms of allergy. No association between latex allergen exposure and symptoms was found in spite of the high frequency of latex allergens. Mite allergens were detected in a minority of rooms. Cat allergen was the most important indoor allergen in the sampled workplace and exposure to this allergen could represent a risk for employees.

Exposure to indoor allergens can occur both at home and in public places such as schools and workplaces. To investigate and compare the presence of indoor allergens in different kind of environments (schools, offices and homes), dust samples were collected from furniture, desks, mattresses and floors with a standardized procedure. Samples were analyzed for Der p 1, Der f 1, Mite group 2 (mites) and Fel d 1(cat) by monoclonal antibody ELISA assay. Mite allergens were detected with low frequencies in schools and workplaces and with high frequency in homes. Fel d 1 was found with high frequency in every examined environment. Homes rather than public places can represent the environment where people can easier incur in mite allergy. All environments could be at risk for cat allergen exposure.

Probiotic bacteria as modulators of the immune response have been intensively studied in reducing the risk of immune-mediated diseases, including atopic diseases. Results from in vitro studies demonstrated that probiotics may modify the polarization of immune cells, supporting potential therapeutic effects in atopic diseases. Several clinical studies have been designed to explore the effective role of probiotics in the modulation of allergic diseases. The results of these studies, although promising, are not conclusive yet and are considered insufficient to recommend probiotics as a part of standard therapy in any allergic conditions. In vivo studies on animal models can provide useful information on the immunologic mechanisms responsible for the potential antiallergic effects of probiotic bacteria. The immunomodulatory activity of the probiotic mixture VSL#3 has been studied in the mouse models of allergic sensitization and anaphylaxis developed in our laboratory with inhalant and food allergens, according to a prophylactic setting by the intranasal route (inhalant allergy model) or a therapeutic setting by the oral route (food allergy model). Intranasally delivered probiotic bacteria prevented the development of Parietaria major allergen-specific response, by down-regulating T helper cell 2 responses at the local and systemic level. Oral therapeutic treatment was able to reduce both systemic and local anaphylactic symptoms induced by oral challenge with the sensitizing allergen Shrimp Tropomyosin. The induction of protective immune responses at the sites of allergen exposure linked to counterregulatory local and systemic immune responses by mucosal delivery of probiotic bacteria mixtures might become an effective strategy in the prevention and therapy of allergic diseases.

T helper type 2 (Th2) cell differentiation requires the expression of GATA-3, a transcription factor that allows transcriptional activation of Th2 cytokine genes through chromatin remodelling. We investigated the role of the negative costimulatory receptor cytotoxic T-lymphocyte antigen 4 (CTLA-4) in the regulation of GATA-3 expression, Th2 differentiation and immunoglobulin production during the immune response to allergens. BALB/c mice were immunized with a recombinant major allergenic component of Parietaria judaica pollen, rPar j I, and treated with blocking anti-CTLA-4 or control antibodies. Results showed that in vivo CTLA-4 blockade enhanced the Par j I-specific immunoglobulin E (IgE) serum level. In contrast, Par j I-specific IgG2a serum level was reduced, suggesting that CTLA-4 blockade skewed immunoglobulin production towards interleukin-4 (IL-4) -dependent immunoglobulin isotypes. Consistently, CTLA-4 blockade increased the frequency of Par j I-specific Th2 cells but not Th1 cells, as well as IL-4 and IL-5 but not interferon-gamma production. Our data also showed that CTLA-4 blockade enhanced the GATA-3 : T-bet messenger RNA ratio. Interestingly, in vivo CTLA-4 blockade did not increase the frequency of GATA-3 protein-expressing cells. In contrast, it enhances GATA-3 protein level per cell. Further, in vitro results show that the anti-CTLA-4 monoclonal antibody, by competing with CD80 for CTLA-4 binding, induced an enhancement in the frequency of IL-4-producing cells that correlates with the increase in GATA-3 protein level per cell. In conclusion, CTLA-4, by affecting the level of GATA-3 per cell, contributes to keeping this factor under the threshold required to become a Th2 effector cell. Consequently, it affects IgE/IgG2a production and contributes to the outcome of allergen-specific immune responses.

Background: Allergy is an immunological disorder affecting about 25% of the population living in the industrialized countries. Specific immunotherapy is the only treatment with a long-lasting relief of allergic symptoms and able to reduce the risk of developing new allergic sensitizations and inhibiting the development of clinical asthma in children treated for allergic rhinitis., Methods: By means of DNA recombinant technology, we were able to design a head to tail dimer expressing disulphide bond variants of the major allergen of the Parietaria pollen. IgE binding activity was studied by Western blot, ELISA inhibition assays and the skin prick test. T cell recognition was studied by peripheral blood mononuclear cell proliferation. The immunogenicity of the hybrid was studied in a mouse model of sensitization., Results: In vitro and in vivo analysis showed that the disruption of specific cysteine residues in both allergens caused a strong reduction in IgE binding activity of the PjEDcys hybrid. In addition,we were able to show that a reduction in the IgE epitope content profoundly reduced the anaphylactic activity of the hybrid (from 100 to 1,000 times less than wild-type allergens) without interfering with the T cell recognition. Sera from BALB/c mice immunized with the hybrid were able to bind the natural Parietaria allergens and to inhibit the binding of human IgE to wild-type Par j 1 and Par j 2 allergens up to 90%., Conclusion: Our results demonstrate that hybrid-expressing disulphide bond variants of the major allergens of the Parietaria pollen displayed reduced allergenicity and maintained T cell reactivity for induction of protective antibodies.

Insects and insect-derived materials have been implicated as a risk factor for sensitization and subsequent elicitation of allergic rhinitis and allergic bronchial asthma. During the last decades, insects other than those known as allergenic, were investigated for their potential role in inducing and triggering an IgE immune response. Among these, the silverfish, an insect belonging to the Thysanura order, appeared to be of particular interest. Silverfish (Lepisma saccharina) is the most primitive living insect, and represents a descendent of the ancestral wingless insects. They are 3-12 mm long, have three tail feelers and are covered with shiny scales. They shun light and need a humid environment and their diet consists of carbohydrate materials such as paper and book-binding glue, crumbs of bread and flour. Because of these features, silverfish finds an optimal habitat both in dwellings and workplaces and in spite of its antiquity, silverfish has succeeded in exploiting the new opportunity created by man. Although its importance significantly increased when it has been demonstrated that house dust contains significant silverfish levels even in houses where the inhabitants were unaware of its presence, no silverfish extract for diagnosis of allergic diseases is commercially available yet. Identification of optimal extraction conditions and characterization of allergenic extracts are the first steps to obtain an effective allergen preparation suitable for diagnosis and therapy, and will be useful as a reference preparation for assessing silverfish exposure in different indoor environments. It has been cloned and characterized a silverfish tropomyosin, named Lep s 1, which represents the first allergen identified in silverfish extract and can be regarded as a molecule cross-reactive among inhalant and edible invertebrates allergenic sources. rLep s 1 displayed biological activity, suggesting that it could be regarded as a useful tool to study the role of silverfish tropomyosin in the sensitization to invertebrate allergic sources.

Purpose: Postoperative pain is the main adverse effect of formal hemorrhoidectomy. A new technique based on Doppler-guided ligation of the terminal branches of the superior hemorrhoidal artery was introduced in 1995 as an alternative to hemorrhoidectomy. The authors report a preliminary experience with this procedure., Methods: The Doppler-guided hemorrhoidal artery ligation technique uses a special proctoscope bearing a Doppler transducer that allows identification and suture ligation of the hemorrhoidal arteries. Sixty-eight consecutive patients (mean age, 48 years; range, 21-74 years) with Grade 3 hemorrhoids were treated., Results: Intraoperative discomfort was measured by a visual analog scale (1-10) and resulted in a mean score of 2.3 (range, 1.3-2.8). Only 38 percent of patients required postoperative analgesia. Patients were examined at 1 week, 1 month, and 3 months and every 6 months thereafter. The mean follow-up was 11 (range, 3-18) months. Bleeding resolved in 91 percent of patients, pain in 73 percent, and prolapse in 94 percent. Complications were recorded in five patients and included persistent pain for more than two days in two patients (3 percent), swelling and thrombosis of one of the hemorrhoids in two patients (3 percent), and a secondary hemorrhage in one patient (1.5 percent)., Conclusion: Doppler-guided ligation of the hemorrhoidal artery is a safe and effective alternative to hemorrhoidectomy and is associated with minimal discomfort and low risk of complications.

Background: Recombinant DNA technology does provide pure, well-defined and reproducible products to be used for clinical purposes, by cloning and expressing the cDNA of allergens present in a specific extract. Ole e 5 is a pollen allergen of Olea europaea with an IgE-binding frequency of about 35%, which has been identified as a superoxide dismutase (SOD). The aim of this study was to clone the cDNA of Ole e 5, to express Ole e 5 in Escherichia coli and to characterize its immunoreactivity., Methods: cDNA of Ole e 5 was amplified by nested 3'-RACE PCR and cloned in pGEX vector 6P expression vector. After sequencing of some clones and homology analysis, the rOle e 5 was produced in an E. coli strain as a fusion protein with GST and purified. Then, the protein immunoreactivity was evaluated by patients' IgE binding (ELISA, ELISA inhibition, and immunoblotting) and by rabbit anti-rOle e 5 binding (immunoblotting and immunoblotting inhibition)., Results: The sequence analysis of Ole e 5 cDNA confirmed that Ole e 5 is a Cu/Zn SOD, with an identity from 90 to 80% with SOD from other species. rOle e 5 was recognized by IgE from 39% of olive pollen-allergic patients tested; moreover, this binding was inhibited by the olive pollen extract. An anti-rOle e 5 antiserum raised in rabbit strongly reacted with a natural component of about 16-kDa molecular weight present in the olive pollen extract; moreover, this binding was inhibited by the recombinant protein., Conclusions: Ole e 5 is the first Cu/Zn SOD identified as an allergen in a pollen source. Due to the widespread presence of this enzyme, rOle e 5 allergen, cloned and expressed in a complete form in E. coli, could represent a good tool to investigate the allergen cross-reactivity between O. europaea pollen and other allergenic sources, such as plant foods and other pollens., (Copyright 2005 S. Karger AG, Basel)

Male breast cancer is rare and accounts for 1% of all breast cancers. The authors report two cases of male breast cancer at Stage III and review the literature. A Madden mastectomy with axillary clearance was performed. Patients were given adjuvant chemotherapy and started on tamoxifen. At one year follow-up the patients are alive and free from disease. Modified radical mastectomy is the preferred surgical approach for localized disease. Adjuvant hormonal therapy with tamoxifen is recommended as first-line treatment. Adjuvant chemotherapy has a role in node-positive cancer and locally advanced disease. Men should be made more aware of the disease and multicentric clinical trials encouraged to ensure an appropriate treatment on the basis of prospectively collected data.

Background: Seborrhoeic dermatitis (SD) is a common chronic inflammatory disease of the skin. Topical steroid creams and/or antifungal agents are commonly used in SD, but no resolutive therapies have been available up to now. Furthermore, little data have been available regarding clinical outcomes after cessation of topical treatments. A new formulation of betamethasone valerate 0.1% in a thermophobic, low-residue, foam vehicle (Bettamousse) (BVM) has become available for the topical treatment of scalp dermatoses. No data have been published hitherto regarding efficacy, safety and patient acceptability of this new formulation for the treatment of SD of the scalp., Study Aim: To assess in an open-label, prospective, multicentre trial, the efficacy, safety and patient acceptability of BVM, as compared to baseline, in SD subjects with scalp involvement., Patients and Methods: A total of 180 patients with moderate-to-severe SD of the scalp were enrolled in the trial. Efficacy was evaluated by analysing SD lesions for erythema, scaling and itching using a five-point grading score (0 = lesion completely cured; 1 = mild; 2 = moderate; 3 = severe and 4 = very severe lesion). The clinical global (sum) score was obtained adding the score of each item. Efficacy and safety were assessed at baseline, after 4 weeks of active treatment, followed by an 8-week follow-up period with no treatment., Results: In comparison with baseline, BVM significantly improved SD lesions. The sum score was reduced from 6.3 +/- 1.8 to 1.4 +/- 1.4 at the end of the active treatment period, (p < 0.0001) and to 1.7 +/- 1.8 at the end of the 8-week follow-up period (p < 0.0001). After active treatment, 93% of the patients had a sum score of

Here we describe the uncommon case of a Caucasian male with secondary hyperparathyroidism due to 8 parathyroid glands discovered in the course of a surgical exploration. The patient (age 49 yr) with a 21-yr history of chronic renal failure came to our observation in June 1999 complaining of depression, muscle weakness, bone and joint pain, movement hindrance. The biochemical evaluation evidenced low-normal serum calcium, high phosphorus and PTH levels. The symptoms and the biochemical findings were suggestive for secondary hyperparathyroidism. The neck US revealed an increase of thyroid gland volume with diffuse hyperechogenity; two nodules of 20 and 25 mm as maximum diameter were found in the thyroid parenchyma, while 4 hypoechogenous nodules (maximum diameter ranging from 13.0 to 30.0 mm) with clean borders and anechogenous areas inside were evidenced in the rear side of the thyroid lobes. The parathyroid scan with 99mTc and 201 Tl demonstrated increased uptake bilaterally in the inferior side of the neck. The patient underwent a total parathyroidectomy with near total thyroidectomy in November 1999. Histological examination of surgical specimen evidenced 6 hyperplastic parathyroid glands in back side of the 2 lobes (3 on the right and 3 on the left), and the examination of the thyroid gland showed 2 hyperplastic parathyroids (5 mm and 15 mm maximum diameter) into the 2 nodules previously evidenced by US. The physiopathological and clinical and therapeutic implications of this observation are discussed.

Background: Calcium-binding plant allergens can be grouped in different families according to the number of calcium-binding domains (EF hands)., Objective: We sought to identify pollens containing crossreactive calcium-binding allergens and to investigate structural and immunologic similarities of members belonging to different families of calcium-binding allergens., Methods: By means of multiple sequence alignment and molecular modeling, we searched for structural similarities among pollen allergens with 2 (Phl p 7, timothy grass; Aln g 4, alder), 3 (Bet v 3, birch) and 4 EF hands (Jun o 4, prickly juniper). Purified recombinant Aln g 4 and Jun o 4 were used to determine the prevalence of IgE recognition in 210 patients sensitized to different pollens and to search, by means of ELISA competition, for the presence of cross-reactive epitopes in pollens from 16 unrelated plant species. IgE cross-reactivity among the allergen families was studied with purified rPhl p 7, rAln g 4, rBet v 3, and rJun o 4 and 2 synthetic peptides comprising the N-terminal and C-terminal EF hands of Phl p 7 by means of ELISA competition., Results: Structural similarities were found by using molecular modeling among the allergens with 2, 3, and 4 EF hands. Pollens from 16 unrelated plants contained Aln g 4- and Jun o 4-related epitopes. Twenty-two percent of the patients with multiple pollen sensitization reacted to at least one of the calcium-binding allergens. A hierarchy of IgE cross-reactivity (rPhl p7 > rAln g 4 > rJun o 4 > rBet v 3) could be established that identified rPhl p 7 as the EF-hand allergen containing most IgE epitopes in the population studied., Conclusion: The demonstration that members of different families of calcium-binding plant allergens share similarities suggests that it may be possible to use representative molecules for the diagnosis and therapy of allergies to EF-hand allergens.

Background: Par j 1 represents a major allergenic component of Parietaria judaica (Pj) pollen, since it is able to induce an immunoglobulin E (IgE) response in 95% of Pj-allergic patients. It belongs to the non-specific lipid transfer protein family, sharing with them a common three-dimensional structure., Methods: Disulphide bond variants of the recombinant Par j 1 (rPar j 1) allergen were generated by site-directed mutagenesis, and the immunological activity of rPar j 1 and its conformational mutants was compared with the use of the skin prick test (SPT). The ability to bind IgE antibodies was evaluated by Western blot, ELISA and ELISA inhibition. T cell reactivity was measured by peripheral blood mononuclear cell proliferation assay., Results: The disruption of Cys14-Cys29 and Cys30-Cys75 bridging (PjA mutant) caused the loss of the majority of specific IgE-binding activity. Additional disruption of the Cys4-Cys52 bridge (PjC mutant) and the latter Cys50-Cys91 bridge (PjD mutant) led to the abolition of IgE-binding activity. On the SPT, PjB (lacking the Cys4-Cys52 and Cys50-Cys91 bridges) was still capable of triggering a type I hypersensitive reaction in 9 out of 10 patients, and PjA in 3 out of 10 patients, while PjC and PjD did not show any SPT reactivity. All the mutants preserved their T cell reactivity., Conclusion: Recombinant hypoallergenic variants of the rPar j 1 allergen described herein may represent a useful tool for improved immunotherapy., (Copyright 2001 S. Karger AG, Basel)

Allergy to Cupressaceae pollen is a worldwide pollinosis caused by several species. Pollen extracts prepared from allergenic species belonging to this family are characterised by low protein and high carbohydrate content. The allergenic components represented in the pollen extracts from different species of the Cupressaceae family show high levels of cross-reactivity when probed with human IgE from allergic subjects and share a number of common epitopes also identified by polyclonal rabbit antisera and monoclonal antibodies. A close relationship has also been described with the Taxodiaceae and Podocarpaceae families. Although both proteic and carbohydrate epitopes appear to be involved in IgE recognition and allergenic cross-reactivity, a large portion of the IgE reactivity of Cupressaceae-allergic patients seems to be associated with sugar moieties present on the relevant allergenic molecules. From this point of view, Cupressaceae/Taxodiaceae allergens constitute a particularly useful model to study IgE cross-reactivity, as they have been shown to display different levels of homology. Moreover, the availability of the purified allergens, together with their recombinant counterparts, may shed light on the actual role played by carbohydrate in allergic sensitisation, IgE recognition and allergenic cross-reactivity., (Copyright 2001 S. Karger AG, Basel)