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Resumen del trabajo presentado en el 8th International Workshop on the Biology of Fish Gametes, celebrado en Gdansk (Polonia) del 20 al 23 de septiembre de 2022., The climate change includes a decrease in seawater pH, and an increase in its temperature. It is possible that the marine fish sperm cells, which are released to the sea at spawning, can be affected by these expected changes on the water, and become unable of fulfill its role on fertilization. For that reason, different seawater pH has been tested in 3 aquaculture marine species: the European eel (Anguilla anguilla), the European seabass (Dicentrarchus labrax), and the Senegalese sole (Solea senegalensis), and their sperm motility parameters analyzed by a CASA-mot system. In the European eel, tested seawater pH, from 6.5 to 9.5, affected sperm motility and other kinetic parameters like MP, FA, VCL, VAP, LIN, STR, WOB, ALH and BFC. pH values lower than 7.8 or higher than 8.2 caused lower values of motility and the rest of kinetic parameters. The longevity was not affected by pH from 7.6 to 8.2. In other experiment a seawater pH of 7.8 caused lower motility, FA, VSL, VAP and LIN than a seawater with pH 8.2. The effect of the water temperature was tested, by comparing activation with seawater at 4 °C (our control) and at 23-24 °C. The spermatozoa beating cross frequency (BCF) was the only parameter significantly affected, being lower at 23 °C than at 4 °C. In other experiment where we combined 2 temperatures (4, 24 °C) and two pH (7.8, 8.2), a significant interaction was observed, and in general the worst results were obtained with seawater at 4 °C and at pH 7.8. Considering all these data, it seems that the seawater pH has a deeper effect on the eel sperm motility than a high temperature. Regarding European sea bass and Senegalese sole, motility and other sperm parameters were not affected by seawater temperature in the pH range from 6.5 to 9.5, thus indicating that ocean acidification would not affect their behavior. The differences found between these species, evolutively and ecologically differents, could reflect different activation mechanisms of the spermatozoa motility., Supported by MCIN with funding from European Union NextGenerationEU (PRTR-C17.I1) and by Generalitat Valenciana (THINKINAZUL/2021/012) to SEASPERM. MM has a postdoctoral contract by Generalitat Valenciana (APOSTD/2020/053). WAG has a Margarita Salas postdoctoral contract (RD 289/2021. UAB) by the Spanish Ministry of Universities. TSF and MPS have fellowships from Brazilian institutions: CNPq (200285/2021-1), and CAPES (Finance code 001), respectively.

Trabajo presentado en la International Conference & Exposition Aquaculture Europe, celebrada en Funchal, Maderia (Portugal) del 04 al 07 de octubre de 2021., [Introduction]: The European eel (Anguilla anguilla) is a highly valued species targeted for aquaculture production. However, it is considered a critically endangered species by CITES, and reproduction in captivity seems to be the only realistic alternative to solve both economic and ecological problems (Asturiano, 2020). It is possible to obtain gametes for artificial fertilization but it requires hormonal treatment, which is a time-consuming and expensive process (Gallego et al., 2012). Moreover, the variable response to these treatments results in high mortality rates during the early stages of development, which could be caused (at least in part) by low sperm quality and uncontrolled epigenetic factors (Herráez et al., 2017). Considering the environmental conditions experienced during eels oceanic migration, the effects of low temperature combined with hormonal treatment have been used to better understand the onset of eel maturation (Peñaranda et al., 2016). Thus, the present study considers the use of low temperature as a pre-treatment before starting a standard hormonal treatment and evaluates its impact on eel maturation and male reproductive performance., [Material and Methods]: Eighty-eight male European eels were maintained for 3-4 days in freshwater at 20 ºC. Then, 6 males were sacrificed, as initial control, and the rest were changed to seawater at 10 ºC, except the control group, which was changed to seawater but maintained at 20 ºC. Low water temperature pre-treatments were applied during 1, 2 or 4 weeks to groups of 20 males. Once finished each pre-treatment, 8 fish per group were sacrificed for sampling. The temperature of the water of the remaining males was increased to 20 ºC in 3-4 days and they started receiving standard hormonal treatment (weekly administration of recombinant chorionic human gonadotropin, rhCG; Ovitrelle, Spain, 1.5 IU/g fish). From sacrificed fish (controls and pre-treated) biometric parameters were measured. Blood and testis samples were used to perform steroid (T, 11-KT and E2) analyses and histology, respectively. Once eels were spermiating, sperm volume, motility and kinetic parameters were evaluated weekly using a CASA system., [Results]: The eye, fin and hepatosomatic indexes from 1-week, 2- and 4-weeks pre-treated fish were significantly higher than those of the control group. The gonadosomatic indexes registered in the control group were unusually high and significantly differed with 1-week pre-treatment group but not with the 2- and 4-weeks groups. Androgen levels (T, 11-KT) were significantly higher after 4-weeks of pre-treatment in comparison with the control. The E2 levels did not show the same increasing profile. The control and 1-week pre-treated males showed significantly higher percentages of undifferentiated and differentiated type A spermatogonia in comparison with early spermatogonia type B. The males after 2- and 4-weeks of pre-treatment revealed a significantly increased proportion of differentiated spermatogonia type A. After the rhCG treatment, males from the 4-weeks pre-treatment group started to produce sperm earlier than males from the other groups. The sperm volume in the 4-weeks pre-treatment group increased during the first weeks of spermiation and in the control group at the end of the experiment, but no significant differences were found. Sperm motility and kinetics parameters were low in pre-treated groups throughout the whole experiment while in the control group increased significantly at the end of the hormonal treatment., [Discussion]: The onset of maturation in the European eel is controlled by environmental conditions found during their migration. The application of pre-treatments can be important to induce maturation in aquaculture eels, which do not encounter these conditions (Palstra and van den Thillart, 2009). Several factors such as temperature, salinity, forced swimming and pressure have been used as part of pre-treatments applied to eels (reviewed by Asturiano et al., 2020). Rozenfeld et al. (2019) evaluated the effects of cold seawater in European eel maturation without hormonal treatment. In eels treated at 10 ºC, the T and 11-KT levels were higher and there was an increase in the proliferation of differentiated type A spermatogonia. Our results also corroborate that low-temperature pre-treatment (4-weeks at 10 °C) induces an early sexual development, thus leading to the release of androgens which promote the proliferation of type A spermatogonia. However, the low sperm quality found in the pre-treated groups revealed that thermal pre-treatment followed by rhCG treatment at high temperatures impairs spermatogenesis. Low-temperature pre-treatments induce testis cell proliferation and synchronization but inhibit further maturation. Peñaranda et al. (2016) suggested that low seawater temperature inhibits the gene expression of enzymes responsible for the synthesis of maturation-inducing steroids (MIS) during eels oceanic migration. Thus, the premature maturation of the testis is prevented before eels arrive to the spawning areas and a temperature threshold is reached allowing the change of the steroidogenic pathway to change from androgens to MIS. In our experiment, lowtemperature pre-treated males had a time-dependent and fast response to rhCG treatment (in comparison with control males maintained at 20 °C), but their spermiation period was shorter and their sperm motility parameters were significantly lower. If this is evidencing a lower rate of gene expression in the testis, and thus limiting the final sperm maturation process, is an open question that requires further research.

Resumen del trabajo presentado en el XIII Congress of the Iberian Association of Comparative Endocrinology, celebrado en modalidad virtual del 16 al 17 de septiembre de 2021., The European eel (A. anguilla) is a valued species targeted for aquaculture production. However, it is considered a critically endangered species and its reproduction in captivity seems to be the only alternative to solve both economic and ecological problems. Although it is possible to obtain eel gametes for artificial fertilization, it requires longterm hormonal treatments. Recent studies found that cold seawater stimulates the brainpituitary-gonad axis in eel males, inducing the start of spermatogenesis. These results suggested that cold seawater could be used as a pre-treatment before the standard hormonal therapy applied to induce sexual maturation, to obtain higher sperm quantity and quality. Thus, our study evaluates the effects of cold seawater pre-treatments of different duration (1, 2 and 4-weeks) on reproductive performance of eel males. Biometric parameters from control and pre-treated fish were measured. Blood and testis samples were collected for steroid (T, 11-KT and E2) and histology analyses, respectively. Once eels were spermiating, sperm parameters were evaluated using a CASA system. Eel males pre-treated with cold temperature showed increased biometric indexes, androgens and E2 plasma levels. Males maintained at 10 ºC for 4-weeks showed a significantly higher percentage of differentiated type A spermatogonia in comparison to undifferentiated ones. However, the low sperm quality found in the pre-treated groups revealed that pretreatments followed by recombinant human chorionic gonadotropin (rechCG) treatment at high temperatures impairs spermatogenesis. Cold temperature pre-treated males had a time-dependent and fast response to rechCG treatment, but their spermiation period was shorter and sperm parameters were significantly lower. Further research is needed to study the effect of thermic pre-treatments on gene regulation controlling spermatogenesis., Funded by the Spanish MICIU (Project EELGONIA; RTI2018-096413-B-I00). LF and MM have contracts funded by Generalitat Valenciana (GRISOLIAP/2020/063 and APOSTD/2020/053). VG had a grant from the MICIU (IJCI2017-34200).

The contribution of coding mutations to oncogenesis has been largely clarified whereas little is known about somatic mutations in noncoding DNA and their role in driving tumors remains controversial. Here, we used an alternative approach to interpret the functional significance of noncoding somatic mutations in promoting tumorigenesis. Noncoding somatic mutations of 151 neuroblastomas (NB) were integrated with ENCODE data to locate somatic mutations in regulatory elements specifically active in NB cells, non-specifically active in NB cells, and non-active. Within these types of elements, transcription factors (TFs) were identified whose binding sites (BS) were enriched or depleted in mutations. For these TFs, a gene expression signature was built to assess their implication in NB. DNA and RNA sequencing data were integrated to assess the effects of those mutations on mRNA levels. The pathogenicity of mutations was significantly higher in TFBS of regulatory elements specifically active in NB cells, as compared to the others. Within these elements, they were 18 over-represented TFs involved mainly in cell cycle phase transitions, and 15 under-represented TFs primarily regulating cell differentiation. A gene expression signature based on over-represented TFs correlated with poor survival and unfavourable prognostic markers. Moreover, recurrent mutations in TFBS of over-represented TFs such as EZH2 affected MCF2L and ADPRHL1 expression, among the others. We propose a novel approach to study the involvement of regulatory variants in NB that could be extended to other cancers and provide further evidence that alterations of gene expression may have relevant effects in NB development.

Microscale fluid dynamics has received intensive interest due to the extraordinary advances in electronic device miniaturization, where the peaks of temperature from hotspots must be reduced by a coolant flowing in a microchannel. One of the most meaningful microscale effects is the emergence of slip flow. The present analysis is concerned with the 2-D velocity distribution of steady-state, hydrodynamically developed, laminar slip flow, for Newtonian fluids in rectangular ducts. The solutions describing velocity profiles, friction factors, shear stresses, momentum flux, and kinetic energy correction factors are derived resorting to the integral transform technique. The results are shown as functions of the aspect ratio and the Knudsen number, in the limit of Kn 0 they reproduce perfectly the well-known results of macroscale fluid dynamics.

We have previously demonstrated that therapeutic concentrations of unfractionated heparin (UFH) impair fibrin polymerization leading to the formation of clots which are more sensitive to lysis induced by tissue plasminogen activator (t-PA). The aim of this study was to compare the effect of UFH with that of three different low molecular weight heparins (LMWHs) on clot sensitivity to t-PA-induced lysis. Labelled fibrin clots, prepared from plasma containing UFH, Fraxiparine, Reviparine, Enoxaparine or saline, were incubated in phosphate buffer containing t-PA (0.1 and 0.5 microgram/ml) and plasminogen (20 micrograms/ml). The extent of clot lysis was quantified by counting the residual radioactivity of the clots and by measuring D-dimer levels released into the medium. Fibrin polymerization and clot structure were evaluated by means of a turbidimetric assay and by electron microscopic scanning. Pre-incubation of plasma with 0.5 or 1.0 U/ml UFH resulted in a marked dose-dependent acceleration of lysis induced by 0.1 or 0.5 microgram/ml t-PA. In contrast, lysis rates induced by 0.5 microgram/ml t-PA were not modified by pre-incubation of plasma with LMWHs. When exposed to 0.1 microgram/ml t-PA clots formed from plasma containing 0.5-2 IU/ml of Fraxiparine, Reviparine and Enoxaparine showed only a minor increase in lysis rates compared to control clots. There was not a clear dose-response curve with LMWHs. Furthermore, lysis rates obtained with UFH-treated clots were always significantly higher than those seen with LMWHs-treated clots.(ABSTRACT TRUNCATED AT 250 WORDS)

Circadian rhythmicity of the structural morphometric model of rat endocrine pancreas has been studied in 24 Wistar female rats, four months old, kept in LD 12:12. The following parameters were evaluated: the volume fractions of nucleus and cytoplasm of exocrine cells, the size distribution and number in unit tissue volume of acinar cell nuclei, the mean nuclear diameter, the shape coefficient of glandular acini (that is the ratio acinar area/perimeter2 which indicates the shifting of structures from circularity). A statistically significant circadian rhythm was demonstrated for the shape coefficient of glandular acini. Results obtained in the present experiment are compared with data recorded in a previous study.

We deal with a family of functionals depending on curvatures and we prove for them compactness and semicontinuity properties in the class of closed and bounded sets which satisfy a uniform exterior and interior sphere condition. We apply the results to state an existence theorem for the Nitzberg and Mumford problem under this additional constraint., Comment: 20 pages. To appear on Rendiconti del Seminario Matematico dell'Universita' di Padova

The process which includes production, collection, carriage and transformation of biomass into renewable fuels and then into energy (both electrical and thermal) involves a large number of decisions to select the most efficient chain. In this paper, the contribution margin of four different energy chains is evaluated both in terms of profits and energy gain per unit of cultivated surface. Four widely cultivated crops in Europe (corn, rapeseed, soybean and sunflower) are considered as feedstock. Four technologies are taken into consideration as conversion plants: (i) anaerobic digestion with internal combustion engines, (ii) direct combustion with Organic Rankine Cycle, (iii) gasification with internal combustion engines, and (iv) straight vegetable oil combustion within internal combustion engines.

Gene transfer delivery of endogenous angiogenesis inhibitors such as angiostatin would circumvent problems associated with long-term administration of proteins. Kaposi's sarcoma (KS), a highly vascular neoplasm, is an excellent model for studying tumor angiogenesis and antiangiogenic agent efficacy. We investigated the effects of angiostatin gene transfer in in vitro and in vivo models of KS-induced neovascularization and tumor growth. A eukaryotic expression plasmid and a Moloney leukemia virus-based retroviral vector for expression of murine angiostatin were generated harboring the angiostatin cDNA with cleavable leader signals under the control of either the strong cytomegalovirus promoter/enhancer or the Moloney leukemia virus long terminal repeat. Angiostatin secretion was confirmed by radioimmunoprecipitation and Western blot analysis. Supernatants of angiostatin-transfected cells inhibited endothelial cell migration in vitro. Stable gene transfer of the angiostatin cDNA by retroviral vectors in KS-IMM cells resulted in sustained angiostatin expression and delayed tumor growth in nude mice, which was associated with reduced vascularization. These findings suggest that gene therapy with angiostatin might be useful for treatment of KS and possibly other highly angiogenic tumors.

One of the aims of the present investigation was to assess three-dimensionally the anteroposterior discrepancy of dental bases using a noninvasive direct procedure. A second aim was to verify the relationship of three-dimensional soft-tissue measurement to the well-established two-dimensional cephalometric assessments of anteroposterior discrepancy. Dental and facial landmarks were directly digitized on 20 orthodontic and maxillofacial surgery patients aged 8 to 26 years using an electromagnetic three-dimensional computerized digitizer. The anteroposterior maxillomandibular discrepancy was measured by calculating the linear distances between the projections of subnasal and sublabial landmarks on the occlusal plane, subnasal and sublabial landmarks on Camper's plane, and insertion of maxillary and mandibular median labial frenula on the occlusal plane. From lateral cephalograms of the same patients, the following measurements were obtained: subspinale point-nasion-supramentale point (ANB) angle; corrected ANB angle that compensates for the position of the maxilla and rotation of the mandible relative to the cranial base; Wits appraisal; MM-Wits, linear distance between the projections of points A and B on the bisector of the palatal plane to mandibular plane angle; and soft-tissue Wits, linear distance between the projections of soft-tissue points A and B on the bisecting occlusal plane. The best two-dimensional vs three-dimensional linear regression (r = 0.91) was found between Wits appraisal and the linear distances between the projections of maxillary and mandibular median labial frenula on the occlusal plane (Wits = -1.05 x 3D measurement - 3.75). The three-dimensional evaluation of the sagittal discrepancy of the jaws directly performed in vivo may allow a more complete analysis of a patient's soft-tissue drape together with the underlying hard-tissue structure.

The thiol N-acetylcysteine (NAC) is a chemopreventive agent that acts through a variety of mechanisms and can prevent in vivo carcinogenesis. We have previously shown that NAC inhibits invasion and metastasis of malignant cells as well as tumor take. Neovascularization is critical for tumor mass expansion and metastasis formation. We investigated whether a target of the anti-cancer activity of NAC could be the inhibition of the tumor angiogenesis-associated phenotype in vitro and in vivo using the potent angiogenic mixture of Kaposi's sarcoma cell products as a stimulus. Two endothelial (EAhy926 and human umbilical vein endothelial [HUVE]) cell lines were utilized in a panel of assays to test NAC ability in inhibiting chemotaxis, invasion, and gelatinolytic activity in vitro. NAC treatment of EAhy926 and HUVE cells in vitro dose-dependently reduced their ability to invade a reconstituted basement membrane, an indicator of endothelial cell activation. Invasion of HUVE cells was inhibited with an ID50 of 0.24 mM NAC, whereas inhibition of chemotaxis required a 10 fold higher doses, indicating that invasion is a preferential target. NAC inhibited the enzymatic activity and conversion to active forms of the gelatinase produced by endothelial cells. The matrigel in vivo assay was used for the evaluation of angiogenesis; NAC strongly inhibited neovascularization of the matrigel sponges in response to Kaposi's sarcoma cell products. NAC prevented angiogenesis while preserving endothelial cells, implying that it could be safely used as an anti-angiogenic treatment.

The thiol N-acetyl-L-cysteine (NAC), an analogue and precursor of reduced glutathione, has cancer chemopreventive properties attributable to its nucleophilicity, antioxidant activity, and a variety of other mechanisms. We demonstrated recently that NAC has anti-invasive, antimetastatic, and antiangiogenic effects in in vitro and in vivo test systems. In the present study, s.c. transplantation of KS-Imm cells in (CD-1)BR nude mice resulted in the local growth of Kaposi's sarcoma, a highly vascularized human tumor. The daily administration of NAC with drinking water, initiated after the tumor mass had become established and detectable, produced a sharp inhibition of tumor growth, with regression of tumors in half of the treated mice along with a markedly prolonged median survival time. The production of vascular endothelial growth factor (VEGF) and certain proliferation markers (proliferating cell nuclear antigen and Ki-67) were significantly lower in Kaposi's sarcomas from NAC-treated mice than from control mice. Treatment of KS-Imm cells with NAC in vitro resulted in a dose-dependent inhibition of chemotaxis and invasion through inhibition of gelatinase-A (matrix metalloproteinase-2, MMP-2) activity without altering MMP-2 or MMP-9 mRNA levels. NAC also significantly inhibited VEGF production but did not affect proliferation markers in vitro. Reverse transcription-PCR analysis indicated that total VEGF mRNAs were reduced by 10 mM NAC. Taken together, these findings provide evidence that NAC, the safety of which even at high doses has been established in almost 40 years of clinical use, in addition to its chemopreventive action, has a strong antiangiogenic potential that could be exploited for preventing cancer progression as well as used in cancer adjuvant therapy.

It is commonly accepted that some form of skin barrier dysfunction is present in canine atopic dermatitis (AD), one of the most common cutaneous pruritic inflammatory diseases of dogs. The impaired skin barrier function facilitates the penetration of allergens and subsequently stronger sensitization responses. The role of the endocannabinoid system (ECS) in the physiology and pathology of the skin is becoming increasingly established. It has been demonstrated that cannabinoid receptors are expressed in healthy and diseased skin and, based on current knowledge, it could be stated that cannabinoids are important mediators in the skin. The present study has been designed to immunohistochemically investigate the expression of the cannabinoid receptors type 1 (CB1R) and 2 (CB2R) and the cannabinoid-related receptors G protein-coupled receptor 55 (GPR55), transient receptor potential vanilloid 1 (TRPV1) and ankyrin 1 (TRPA1), peroxisome proliferator-activated receptors alpha (PPARα), and serotoninergic receptor 1a (5-HT1aR) in keratinocytes of healthy dogs and of dogs with AD. Samples of skin tissues were collected from 7 healthy controls (CTRL-dogs) and from 8 dogs with AD (AD-dogs). The tissue samples were processed using an immunofluorescence assay with commercially available antibodies, and the immunolabelling of the receptors studied was quantitatively evaluated. The keratinocytes of the CTRL- and the AD-dogs showed immunoreactivity for all the receptors investigated with a significant upregulation of CB2R, TRPA1, and 5-HT1aR in the epidermis of the AD-dogs. The presence of cannabinoid and cannabinoid-related receptors in healthy keratinocytes suggested the possible role of the ECS in canine epidermal homeostasis while their overexpression in the inflamed tissues of the AD-dogs suggested the involvement of the ECS in the pathogenesis of this disease, having a possible role in the related skin inflammation and itching. Based on the present findings, the ECS could be considered a potential therapeutic target for dogs with AD.

The aim of this retrospective single-center study was to evaluate which factors, including expression of P-glycoprotein (P-gp), a membrane-bound protein involved in multiple drug resistance, could predict the response to treatment in canine immunosuppressant-responsive enteropathy (IRE). Dogs with IRE or non-responsive enteropathy (NRE) that were examined from 2005 to 2014 were included and were divided into two groups (IRE vs. NRE). Signalment, history, and clinical and laboratory findings were collected. P-glycoprotein immunohistochemistry was carried out on duodenal biopsies of both groups stored in our biobank, and immunophenotyping and molecular clonality were performed on the NRE samples. Ninety-two dogs were enrolled, 73 IRE (79.3%) and 19 NRE (20.7%), with a prevalence of pure breed (78.3% vs. 21.7%) and male dogs (p &lt, 0.001). Factors associated with a worse prognosis were previous treatment with steroids (p = 0.033) and lower serum total protein concentration (p = 0.005). Clonality testing on the NRE duodenal biopsies showed 5/16 clonal responses, assuming a latent undiagnosed lymphoma as a possible cause of the NRE.

Feline Vector-Borne Diseases show increased global prevalence and some Anaplasma and Ehrlichia species may pose a risk to human health. The diagnosis of Anaplasma and Ehrlichia species infection in cats is achieved by the combined use of different methods as cytologic examination evidencing intracytoplasmic morulae, serologic tests and molecular assays. The peripheral whole blood is considered the sample of choice for Anaplasma and Ehrlichia species DNA detection in cats, but false negative results are reported leading to underestimation of infection prevalence. In order to have a more accurate assessment of the spread of feline vector-borne pathogens, the presence of Anaplasma spp. and Ehrlichia spp. DNA in 37 owner and shelter cats subjected to necropsy were prospectively investigated by testing in end-point PCR spleen, bone marrow, blood clot and hair samples. The bacteria identified were genetically characterised. Three shelter cats tested positive for A. phagocytophilum DNA in spleen (one cat) or in hair samples (two cats). None of the cats tested positive in bone marrow and blood samples. From the results obtained, it can be assumed that the use of spleen or hair samples could allow a more reliable detection of A. phagocytophilum DNA in cats with blood tested negative. In the phylogeny constructed with a fragment of the heat shock (groEL) gene nucleotide sequences, all the identified A. phagocytophilum clustered with bacteria infecting a wide range of hosts, including humans, showing a potential zoonotic role.

Neuroblastoma (NB) is the most common extracranial malignant tumor of childhood and is characterized by a broad heterogeneity in clinical presentation and evolution. Recent advances in pangenomic analysis of NB have revealed different recurrent chromosomal aberrations. Indeed, it is now well established that the overall genomic profile is important for treatment stratification. In previous studies, 11 genes were shown to be recurrently amplified (ODC1, ALK, GREB1, NTSR2, LIN28B, MDM2, CDK4, MYEOV, CCND1, TERT, and MYC) besides MYCN, with poor survival of NB patients harboring these amplifications being suggested. Genomic profiles of 628 NB samples analyzed by array-comparative genome hybridization (a-CGH) were re-examined to identify gene amplifications other them MYCN amplification. Clinical data were retrospectively collected. We additionally evaluated the association of FRS2 gene expression with NB patient outcome using the public R2 Platform. We found eight NB samples with high grade amplification of one or two loci on chromosome arm 12q. The regional amplifications were located on bands 12q13.3-q14.1 and 12q15-q21.1 involving the genes CDK4, MDM2, and the potential oncogenic gene FRS2. The CDK4, MDM2, and FRS2 loci were coamplified in 8/8 samples. The 12q amplifications were associated with very poor prognosis and atypical clinical features of NB patients. Further functional and clinical investigations are needed to confirm or refute these associations.

Background:Quinidine at high dose is suggested as antiarrhythmic treatment in patients with Brugada Syndrome (BrS), but its efficacy to prevent life-threatening arrhythmic events (LAE) in this population is unproven and its use limited by frequent side effects. We assessed whether low-dose quinidine in patients with BrS reduces the following: (1) the occurrence of LAEs at follow-up, as compared to no therapy, in a population of both high-risk survivors of LAE and lower risk patients asymptomatic for LAEs and (2) the arrhythmic burden in the high-risk group of cardiac arrest survivors.Methods:We compared the clinical course of 53 patients with BrS treated with quinidine to that of 441 untreated controls, matched by sex, age, symptoms, and the duration of observation. Furthermore, we calculated the annual incidence of LAE off-quinidine and on-quinidine in 123 patients with BrS who were cardiac arrest survivors.Results:Fifty-three patients with BrS (89% males, median age 39.8 years) received quinidine (439±115 mg/d) for 5.0±3.7 years. Therapy was stopped in 3 cases (6%) for side effects. Quinidine reduced by 26% the risk of experiencing an LAE in cases versus controls (hazard ratio, 0.74; 95% CI, 0.22–2.48;P=0.62). Furthermore, in 27 of 123 patients with BrS symptomatic for LAEs who were treated for 7.0±3.5 years, the annual rate of LAEs decreased from 14.7% while off-quinidine to 3.9% while on-quinidine (P=0.03). Of note, recurrent LAEs were recorded in 4 (15%) cardiac arrest survivors while on-quinidine.Conclusions:We demonstrated for the first time in the long-term that low-dose quinidine reduces recurrent LAEs in patients with BrS who had already survived an LAE, with few side effects. Remarkably, 15% of patients symptomatic for LAEs experience recurrent life-threatening arrhythmias while on-treatment, suggesting that quinidine cannot replace implantable defibrillators in high-risk subjects.

The diffusion of district heating (DH) technologies is closely related to the improvement in design and management techniques. Holistic modeling of DH networks represents one of the most promising tools in this regard, since it is aimed at both identifying the size of new plants and optimizing existing ones, with a significant reduction in the time, costs and complexity of real-field tests. The aim of this work is to present the application of two innovative modeling procedures to a real case, i.e. a branch of the Parma University Campus network with a length of about 2 km, 12 connected buildings and an overall heating requirement of 11.2 MW. The first procedure is based on a dedicated library of energy system components developed by the authors in the Matlab®/Simulink® environment, which can be used to assemble models of systems and grids following different configurations. Power generation, conversion and distribution systems together with buildings and generic users can be simulated, considering the thermal and pressure losses of the heat transfer fluid. Typical disturbances can be considered, e.g. solar irradiance, environmental temperature variations and metabolic heat gain from people. Furthermore, a realistic control strategy has been applied to valve opening and boiler operation, in order to fulfil the heat demand profile of buildings in terms of indoor temperatures. Results show that the simulation tool provides a detailed representation of the real system behavior and allows for a comprehensive analysis of the evolution of the main thermodynamic parameters of the heat transfer fluid. The second procedure is based on a tool able to simulate a complex energy distribution network considering the contemporary flows of electrical, thermal and cooling energy. This approach allows both the energy distribution and its generation to be optimized by adopting an optimization procedure based on a genetic algorithm that allows for the load allocation of all the generation systems minimizing the costs of energy production and the associated environmental impact. The results of this second approach, applied to the case study of the Parma University Campus network, show a decrease in electricity consumption for pumping coupled with a reduction in CO2 emissions. The two approaches allow a complete description of a complex smart grid by considering all its aspects from the optimal load distribution of the energy systems up to the implementation of control strategies for the management of the room temperatures at the final users.The diffusion of district heating (DH) technologies is closely related to the improvement in design and management techniques. Holistic modeling of DH networks represents one of the most promising tools in this regard, since it is aimed at both identifying the size of new plants and optimizing existing ones, with a significant reduction in the time, costs and complexity of real-field tests. The aim of this work is to present the application of two innovative modeling procedures to a real case, i.e. a branch of the Parma University Campus network with a length of about 2 km, 12 connected buildings and an overall heating requirement of 11.2 MW. The first procedure is based on a dedicated library of energy system components developed by the authors in the Matlab®/Simulink® environment, which can be used to assemble models of systems and grids following different configurations. Power generation, conversion and distribution systems together with buildings and generic users can be simulated, considering the th...

There is little information on sequence variation of canine adenovirus type 1 (CAdV-1), the aetiological agent of infectious canine hepatitis (ICH). This study reports hexon and fibre gene sequence variants of CAdV-1 in a dog with systemic ICH and a dog with the ocular form of the disease (‘blue eye’) in Northern Italy in 2013. One of the sequence variants matched a CAdV-1 fox sequence previously detected in Italy.

In recent years, cogeneration systems have gained increasing attention especially when dealing with distributed generation for residential buildings. One of the main problems with using cogenerative systems in residential building applications is that the demand for heat and electricity is not synchronized. For this reason, when the combined heat and power system operates during electricity peak hours (i.e. the rate of the electricity is higher), it could be profitable to store the heat in order to satisfy delayed demands. This paper presents a model for the calculation of the profitability of micro combined heat and power systems for residential building applications. The model takes into account hourly demands calculated by means of monthly and daily load profiles for heat and electricity. The system under consideration is composed of a CHP system, an auxiliary boiler and a heat-storage tank. The model is applied to a single-family dwelling in order to evaluate the effect of the size of the thermal energy storage unit on the energy and economic performance of four different prime movers (an internal combustion engine, a Stirling engine, a micro Rankine cycle and a thermophotovoltaic system). Thermal energy produced, electrical energy produced, self-consumed or exchanged with the grid, consumed natural gas, as well as differential cash flow with respect to separate generation and payback period are presented. The effect of the size of the thermal energy storage proves to be not linear with respect to the thermal power of the prime mover.

A 12-year-old, neutered male, mixed-breed, domestic cat was presented for dysuria and haematuria accompanied by recurrent dyschezia and constipation. At rectal digital examination a non-painful, globose and relatively immobile mass was detected at the pelvic brim, ventral to the rectum. Abdominal ultrasound showed the prostate replaced by a mass with a regular shape but patchy/non-uniform echogenicity. A prostatectomy followed by prepubic urethrostomy was performed. At histopathology the tumour was composed of epithelial ducts and acini with malignant features, surrounded by proliferating cellular stroma with mitoses and nuclear atypia; an immunohistochemical panel confirmed the biphasic nature of the tumour proliferation. The diagnosis was non-infiltrating malignant mixed tumour, resembling human sarcomatoid carcinoma of the prostate. Two years after surgery the animal has not shown any health problems. To the author's knowledge, this is the first reported case of prostatic tumour with a positive follow-up after surgical treatment in cat.

In this paper some new tools for the study of evolution problems in the framework of Young measures are introduced. A suitable notion of time-dependent system of generalized Young measures is defined, which allows to extend the classical notions of total variation and absolute continuity with respect to time, as well as the notion of time derivative. The main results are a Helly type theorem for sequences of systems of generalized Young measures and a theorem about the existence of the time derivative for systems with bounded variation with respect to time., Comment: 32 pages

The aim of the study was to compare, in 49 dogs with clinical and histopathological diagnosis of IBD, the endoscopic features (transforming the qualitative variables in quantitative ones by application of new codes), serum folate values, serum cobalamin values and the average number of T CD3 lymphocytes/500 μm in duodenal epithelial layer and the average number of T CD3 lymphocytes/10000 μm2 in duodenal lamina propria. A significant correlation (p