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ABSTRACT: Japanese quails in wild life live in small groups with females being even solitary during the laying period. Although it is a poultry species widely used for egg production, information regarding laying behavior motivations or influencing variables is scarce. Our study focuses on evaluating along 7 d the quail laying behavior in a novel environmental set up. This set up allows the female to choose between remaining separated from a conspecific in one side of the apparatus or to voluntarily enter their space (box-mate side) and interact with it. We evaluated whether the female insemination status prior to enter the environmental set up, and the presence of a female or a male partner in the box-mate side can influence their laying and social behavior. Thus, 4 experimental groups were established. Females spent a higher (P < 0.05) percentage of time in the box-mate side than in their separated sector in all groups. In 3 of the 4 experimental groups (non-inseminated females interacting with a female or a male box-mate, and inseminated females interacting with a male box-mate) females also laid a greater percentage (≥65%, P < 0.05, in all cases) of eggs in the box-mate sector than in their separated sector. However, the group of inseminated females that interacted with a female box-mate shifted their egg distribution and laid equally between both sides of the apparatus. Aggressive social interactions were reduced (P < 0.05) throughout the testing days but this was depending upon the female insemination status and the sex of their box-mate. Findings suggest that females can change their laying side choice when they are inseminated but depending on the sex identity of their box-mate partners. Thus, providing quail female breeders with the option of laying their eggs in separated enclosures from conspecifics could be key to favor their well-being.

Japanese quails in wild life live in small groups with females being even solitary during the laying period. Although it is a poultry species widely used for egg production, information regarding laying behavior motivations or influencing variables is scarce. Our study focuses on evaluating along 7 d the quail laying behavior in a novel environmental set up. This set up allows the female to choose between remaining separated from a conspecific in one side of the apparatus or to voluntarily enter their space (box-mate side) and interact with it. We evaluated whether the female insemination status prior to enter the environmental set up, and the presence of a female or a male partner in the box-mate side can influence their laying and social behavior. Thus, 4 experimental groups were established. Females spent a higher (P < 0.05) percentage of time in the box-mate side than in their separated sector in all groups. In 3 of the 4 experimental groups (non-inseminated females interacting with a female or a male box-mate, and inseminated females interacting with a male box-mate) females also laid a greater percentage (≥65%, P < 0.05, in all cases) of eggs in the box-mate sector than in their separated sector. However, the group of inseminated females that interacted with a female box-mate shifted their egg distribution and laid equally between both sides of the apparatus. Aggressive social interactions were reduced (P < 0.05) throughout the testing days but this was depending upon the female insemination status and the sex of their box-mate. Findings suggest that females can change their laying side choice when they are inseminated but depending on the sex identity of their box-mate partners. Thus, providing quail female breeders with the option of laying their eggs in separated enclosures from conspecifics could be key to favor their well-being., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: The incidence of cutaneous melanoma is increasing in Italy, in parallel with the implementation of gene panels. Therefore, a revision of national genetic assessment criteria for hereditary melanoma may be needed. The aim of this study was to identify predictors of susceptibility variants in the largest prospective cohort of Italian high-risk melanoma cases studied to date., Materials and Methods: From 25 Italian centers, we recruited 1044 family members and germline sequenced 940 cutaneous melanoma index cases through a shared gene panel, which included the following genes: CDKN2A, CDK4, BAP1, POT1, ACD, TERF2IP, MITF and ATM. We assessed detection rate according to familial status, region of origin, number of melanomas and presence and type of non-melanoma tumors., Results: The overall detection rate was 9.47% (5.53% analyzing CDKN2A alone), ranging from 5.14% in sporadic multiple melanoma cases (spoMPM) with two cutaneous melanomas to 13.9% in familial cases with at least three affected members. Three or more cutaneous melanomas in spoMPM cases, pancreatic cancer and region of origin predicted germline status [odds ratio (OR) = 3.23, 3.15, 2.43, P < 0.05]. Conversely, age > 60 years was a negative independent predictor (OR = 0.13, P = 0.008), and was the age category with the lowest detection rate, especially for CDKN2A. Detection rate was 19% when cutaneous melanoma and pancreatic cancer clustered together., Conclusions: Gene panel doubled the detection rate given by CDKN2A alone. National genetic testing criteria may need a revision, especially regarding age cut-off (60) in the absence of strong family history, pancreatic cancer and/or a high number of cutaneous melanomas., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background Aims: Induced pluripotent stem cells (iPSCs) have the capacity to generate β cells in vitro, but the differentiation is incomplete and generates a variable percentage of off-target cells. Single-cell RNA sequencing offers the possibility of characterizing the transcriptional dynamics throughout differentiation and determining the identity of the final differentiation product., Methods: Single-cell transcriptomics data were obtained from four stages across differentiation of iPSCs into β cells and from human donor islets., Results: Clustering analysis revealed that iPSCs undertake a full endoderm commitment, and the obtained endocrine pancreatic cells have high homology with mature islets. The iPSC-derived β cells were devoid of pluripotent residual cells, and the differentiation was pancreas-specific, as it did not generate ectodermal or mesodermal cells. Pseudotime trajectory identified a dichotomic endocrine/non-endocrine cell fate and distinct subgroups in the endocrine branch., Conclusions: Future efforts to produce β cells from iPSCs must aim not only to improve the resulting endocrine cell but also to avoid differentiation into non-pancreatic endoderm cells., (Copyright © 2020 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.)

The blockade of pro-inflammatory mediators is a successful approach to improve the engraftment after islet transplantation. L-aptamers are chemically synthesized, nonimmunogenic bio-stable oligonucleotides that bind and inhibit target molecules conceptually similar to antibodies. We aimed to evaluate if blockade-aptamer-based inhibitors of C-C Motif Chemokine Ligand 2/monocyte chemoattractant protein-1 (CCL2/MCP-1) and C-X-C Motif Chemokine Ligand 12/stromal cell-derived factor-1 (CXCL12/SDF-1) are able to favor islet survival in mouse models for islet transplantation and for type 1 diabetes. We evaluated the efficacy of the CCL2-specific mNOX-E36 and the CXCL12-specific NOX-A12 on islet survival in a syngeneic mouse model of intraportal islet transplantation and in a multiple low doses of streptozotocin (MLD-STZ) diabetes induction model. Moreover, we characterized intrahepatic infiltrated leukocytes by flow cytometry before and 3 days after islet infusion in presence or absence of these inhibitors. The administration for 14 days of mNOX-E36 and NOX-A12 significantly improved islet engraftment, either compound alone or in combination. Intrahepatic islet transplantation recruited CD45 + leucocytes and more specifically CD45+/CD11b+ mono/macrophages; mNOX-E36 and NOX-A12 treatments significantly decreased the recruitment of inflammatory monocytes, CD11b + /Ly6C high /CCR2 + and CD11b + /Ly6C high /CXCR4 + cells, respectively. Additionally, both L-aptamers significantly attenuated diabetes progression in the MLD-STZ model. In conclusion, CCL2/MCP-1 and CXCL12/SDF-1 blockade by L-aptamers is an efficient strategy to improve islet engraftment and survival., (© 2019 The American Society of Transplantation and the American Society of Transplant Surgeons.)

Background: Bone marrow (BM) is as an alternative site for islet transplantation, but it is not an immunoprotected microenvironment and allogeneic islets are rejected. However, the BM, for its structure and anatomic position, offers the possibility to modulate microenvironment by local interventions. We here investigate whether local irradiation is able to improve islet engraftment and prevent rejection in BM in the absence of immunosuppression., Methods: A model of BM local irradiation was set up. Islets were transplanted in syngeneic and fully major histocompatibility complex-mismatched recipients in control and locally irradiated BM; gain of normoglycemia and time to rejection were evaluated., Results: BM local irradiation proved to be a selective and safe procedure. Syngeneic islet transplantation into locally irradiated BM had better outcome compared with not irradiated recipients in terms of capacity to gain normoglycemia (100% versus 56% in irradiated versus not irradiated mice). In the allogenic setting, glycemia was significantly lower in the first days after transplantation in the group of irradiated mice and local irradiation also delayed time to graft rejection (from 4 ± 1 days for not irradiated to 11 ± 1 days for locally irradiated mice)., Discussion: These data indicate that local immunosuppression by irradiation before islet transplantation in BM favors islet engraftment and delays time to rejection., (Copyright © 2019 International Society for Cell and Gene Therapy. Published by Elsevier Inc. All rights reserved.)

Consumer concern on the quality of products and animal welfare has greatly increased during the past decades. Dietary synthetic antibiotic products used as growth promoters have been restricted or banned in many countries. Edible plants, essential oils, or their main components were suggested as natural feed supplements to improve growth, products' quality, and welfare-related parameters. Thymol (THY), a main component of oregano essential oil, has been proved as an effective antimicrobial and antioxidant compound. Tocopherol (TOC) evidenced antioxidant activity with potential as a growth promoter and a synergic antioxidant activity between TOC and ascorbyl palmitate (AP) has also been reported. Herein, we evaluated whether broiler diet supplementation with THY, and THY with a formulation mix containing TOC and AP (1:0.5:0.5, respectively) have potential as growth enhancers under commercial conditions. Potential protective effects against foot pad dermatitis and hock burns were also evaluated. Newly hatched male broiler chicks with similar body weight (BW) were randomly assigned to 1 of 7 groups (4 replicates each) as follows: Basal (no feed supplements added), Promotor (Basal + 6.26 μmol flavomycin/kg feed), BHT (Basal + 1.33 mmol of buthylated hidroxytoluene (BHT)/kg feed), Prom-BHT (Basal + 6.26 μmol flavomycin/kg feed + 1.33 mmol of BHT/kg feed), TOC-AP (Basal + 0.67 mmoles of TOC + 0.67 mmoles of AP/kg feed), THY (Basal + 1.33 mmoles of THY/kg feed), and THY-TOC-AP (Basal + 0.67 mmoles of THY + 0.67 mmoles of a mix 1:1 of TOC-AP). Along 7 wk, BW, feed intake, and feed conversion ratio were evaluated. Skin injuries were assessed at 35 d of age. At the end of the study (42 d), compared to Basal group, similarly enhanced final BW were observed in all groups but TOC-AP. No main differences between groups were detected in feed intake, feed conversion ratio, or skin injuries. Findings suggest that THY itself or in combination with TOC-AP may have value as a natural growth enhancer alternative for broilers., (© 2018 Poultry Science Association Inc.)

Aggressive behaviors can affect both animal welfare and productivity. Because the expression of aggressive behaviors is dependent on the quality of the opponent, they reflect relative rather than absolute levels of underlying aggressiveness. This study was aimed to characterize the aggressive responsiveness of photostimulated (14:10 h light:dark photoperiod) adult Japanese quail when interacting with a photocastrated (6:18 h light:dark photoperiod) counterpart in a novel test environment and to assesses interindividual variations. This was based on the assumption that photocastrated birds will not actively provoke an aggressive confrontation. Birds were reared in male-female pairs. Frequencies of behaviors (i.e., pecks, threats, chases, grabs, mounts) were determined during 10 min social interactions in a novel environment. A first experiment evaluated 78 encounters between a photostimulated male or female with either a photocastrated male or female (photocastration of sexually mature birds started at 11 wk of age). High interindividual variability was observed and in general, highly aggressive birds (performing 20 or more aggressive interactions) received little or no aggression from their test counterpart. However, unexpectedly, we also found that 37% and 32% of photocastrated males and females, respectively, performed aggressions toward their photostimulated counterparts, and initiated the aggressive interactions in a similar proportion than photostimulated males. Aggressive photocastrated males did not perform reproductive-type behaviors (i.e., grabs, mounts). Aggressiveness in the photocastrated birds was attributed to their social experience prior to photocastration. Therefore, a second experiment evaluated 106 encounters between a photostimulated male or female and a naive photocastrated male (photocastration started at 4 wk of age, prior to sexual development). Photocastrated males performed no aggressions toward their photostimulated counterparts. Consistently with previous studies, our findings show that naive photocastrated males can be used as a non-aggressive stimulus during a social interaction aimed to assess expression of aggressiveness in photostimulated birds. However, caution should be taken when applying the photocastration protocol considering that prior fighting and sexual experience or other physiological changes related with maturation can interfere during subsequent aggressive testing., (© 2017 Poultry Science Association Inc.)

Iatrogenic pancreatic cancer metastasis after islet infusion is a potential risk of islet autotransplantation performed after pancreatectomy. To model this risk, islets and/or pancreatic exocrine clusters obtained from a genetically engineered mouse model for pancreatic ductal adenocarcinoma (the LSL-Kras G12D/+ ;LSL-Trp53 R172H/+ ;Pdx-1-Cre, termed KPC mouse) were transplanted via the portal vein in syngeneic wild type (WT) severely diabetic recipients in the following treatment groups: group A (n = 11) received KPC exocrine clusters in volume equal to 250 islet equivalents (IEQs); group B (n = 12) received 250 WT IEQs mixed with KPC exocrine clusters (1:1 volume ratio); group C (n = 5) received 250 KPC IEQs, and group D (n = 7) received 250 WT IEQs. The incidence of hepatic metastasis was assessed by magnetic resonance imaging and histology over the 13 months of follow-up. Overall survival was not different in the four groups. No mice developed liver metastases during the follow-up. Two mice developed spontaneous tumors: a liver hepatocellular tumor in group A and a malignant lymphoma in group D. Islets and/or exocrine clusters obtained by KPC mouse, a model that develops pancreatic cancer with 100% penetrance, do not retain the same risk of tumor development when transplanted via the portal vein in a syngeneic diabetic recipient., (© 2017 The American Society of Transplantation and the American Society of Transplant Surgeons.)

Background: The advancement of knowledge in the field of regenerative medicine is increasing the therapeutic expectations of patients and clinicians on cell therapy approaches. Within these, stem cell therapies are often evoked as a possible therapeutic option for diabetes, already ongoing or possible in the near future., Aim: The purpose of this document is to make a point of the situation on existing knowledge and therapies with stem cells to treat patients with diabetes by focusing on some of the aspects that most frequently raise curiosity and discussion in clinical practice and in the interaction with the patient. In fact, at present there are no clinically approved treatments based on the use of stem cells for the treatment of diabetes, but several therapeutic approaches have already been evaluated or are being evaluated in clinical trials., Data Synthesis: It is possible to identify three large potential application fields: 1) the reconstruction of the β cell mass; 2) the immunomodulation in type 1 diabetes (T1D); 3) the treatment of complications. In this study we will limit the discussion to approaches that have the potential for clinical translation, deliberately omitting aspects of basic biology and preclinical data. Also, we intentionally omit the treatment of the complications that will be the subject of a future document. Finally, an overview of the Italian situation regarding the storage of cord blood cells for the therapy of diabetes will be given., (Copyright © 2017 The Italian Society of Diabetology, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition, and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.)

Background: In vitro primary cultures of microvascular endothelial cells from endocrine pancreas are difficult to obtain, but can be a very helpful tool for studies of islet biology, transplantation and regenerative medicine., Methods: We applied a protocol recently described for the isolation and culture of brain microvascular endothelial cells (EC) on human pancreatic islets. EC obtained were characterized in terms of morphological (light and transmission electron microscopy), phenotypical (by immunofluorescence and flow cytometry) and functional (cord formation assay and protein secretion by multiplex bead-based assay) characteristics., Results: EC were obtained from 25% of islet preparations processed. Two primary endothelial cell lines showed high proliferative potential and were deeply characterized: they presented endothelial cell morphology and expressed CD31, CD49a, CD49e, CD34, von Willebrand Factor (vWF), Vascular Endothelial CAdherin (VE-CAD), Tyrosine Kinase with Ig and EGF Homology Domains-2 (TIE2), Vascular Endothelial Growth Factor Receptor 1 (VEGFR1), Ulex lectin and the endothelium endocrine-specific marker nephrin. Besides, they were able to form cordons in vitro and secreted factors involved in the process of angiogenesis such as Vascular Endothelial Growth Factor (VEGF), Monocyte Chemotactic Protein 1 (MCP-1), interleukin (IL)-8 and Melanoma Growth Stimulatory Activity Alpha (GROα). These cell lines were termed Human Islet Microvascular Endothelial Cells (HIMEC)., Discussion: This study establishes a simple and effective strategy for isolation and long-term culture of EC derived from human pancreatic islet. HIMEC in culture preserve phenotype and functional properties and are, therefore, a useful tool for future experiments of in vitro pancreas modelling, co-transplantation with pancreatic islets, re-vascularization of scaffold or matrix for regenerative medicine purposes., (Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)

Social interactions have been extensively studied in poultry in a variety of environmental situations. Many studies allow full social contacts between birds, but there are others in which the interactions are tested through barriers (wire mesh or glass). Thus a situation where, according to their needs, some birds can get access to physical contact with conspecifics while others cannot, would be useful to expand the testing options for social interaction studies. We developed an individual physical barrier device (IPB) that is fitted on the birds to delimit their ambulation areas by preventing them from passing across metal mesh boundaries that IPB free counterparts can easily overcome. The prototypes showing greater efficacy consisted of a small metal bar placed in the bird's back perpendicular to the sagittal plane that slightly exceeds body width, held with a harness fitted by 2 elastic fabric bands around the wings' base. To be useful, the IPB should allow natural movements and not affect the expression of behaviors (non-invasive). This study assessed whether the IPB may alter adult Japanese quail behavioral responses using 4 classical test situations: Open-Field, Runway, Time Budget in Home Box, and Mating Interactions. Open-field ambulatory behaviors were affected 1 h, but not 7 d, after IPB was fitted, suggesting that 7 d (or less) are required to habituate to the device. After that time period, IPB fitted birds showed no differences in any of the behaviors registered in the other 3 tests situations when compared to non IPB fitted birds. Findings suggest that after habituation, the IPB does not affect main behaviors in adult quail. Its application could be expanded if an IPB device is also found suitable for other poultry species., (© 2015 Poultry Science Association Inc.)

The use of noninvasive methods for measuring fecal glucocorticoid metabolites is a useful tool for endocrine assessment particularly in studies where animals cannot be captured, when they should be sampled without disturbing their activities, and/or when welfare needs to be maximized. However, still no complete standardization exists for the methodology, and some confounding variables may play an important role affecting measurements and interpretation of results. The present study focused on whether two different diets (laying feed or seed mixture), the time since defecation (0, 4, 24, or 48 h) and the drying method of those samples (oven-dried or naturally nonoven-dried) may affect concentrations of corticosterone metabolites (CM) measured in male Japanese quail. Half of the birds were provided with plain water (control) and the other half received a corticosterone solution. Birds fed with a seed mixture exhibited higher values of CM (nanogram/gram) in droppings than quail that received a laying feed diet suggesting that diet should be carefully considered as a potential source of variation. As expected both groups exhibited higher CM concentration after corticosterone treatment. While CM concentrations increased significantly in nonoven-dried samples over time (0 < 4 < 24 = 48 h), oven-dried samples exhibited similar high CM values. At 24 and 48 h postdefecation, nonoven-dried samples had similar CM concentrations as all oven-dried samples. Drying of samples may be considered a reliable method to reduce variations due to water loss over time, facilitating comparisons up to 48 h postdefecation. This finding would allow to enhance the range of application of this noninvasive and welfare friendly method to situations where samples cannot be collected or frozen shortly after defecation., (© 2015 Poultry Science Association Inc.)

In order to broaden knowledge on the pathogenesis of malignant pleural mesothelioma (MPM), we reviewed studies on the MPM-transcriptome and identified 119 deregulated genes. However, there was poor consistency among the studies. Thus, the expression of these genes was further investigated in the present work using reverse transcriptase-quantitative PCR (RT-qPCR) in 15 MPM and 20 non-MPM tissue samples. Fifty-nine genes showed a statistically significant deregulation and were further evaluated in two epithelioid MPM cell lines (compared to MET-5A, a non-MPM cell line). Nine genes (ACSL1, CCNO, CFB, PDGFRB, SULF1, TACC1, THBS2, TIMP3, XPOT) were deregulated with statistical significance in both cell lines, 12 (ASS1, CCNB1, CDH11, COL1A1, CXADR, EIF4G1, GALNT7, ITGA4, KRT5, PTGIS, RAN, SOD1) in at least one cell line, whereas 7 (DSP, HEG1, MCM4, MSLN, NME2, NMU, TNPO2) were close but did not reach the statistical significance in any of the cell line. Patients whose MPM tissues expressed elevated mRNA levels of BIRC5, DSP, NME2, and THBS2 showed a statistically significant shorter overall survival. Although MPM is a poorly studied cancer, some features are starting to emerge. Novel cancer genes are suggested here, in particular those involved in cell-cell and cell-matrix interactions., (Copyright © 2014 Elsevier B.V. All rights reserved.)

One challenge faced by ethologists in poultry welfare is helping birds to form functionally successful groups over time through the expression of appropriate behaviors. Searching for phenotypic variability, we developed in Japanese quail a density-related permanence (DRP) test that enables us to classify young birds (while in groups) according to their individual permanence in boxes containing a high or low density of confined conspecifics (HD or LD, respectively). This study addressed the question of whether contrasting DRP quail behavior may reflect underlying differences in social responses. Birds were classified at 11 d of age in an apparatus consisting of 2 boxes interconnected by a central region delimited by sliding doors. Each box contained at its distal end either 12 or 3 conspecifics confined behind a glass (high or low density, respectively). The doors were closed 9 times every 1 h, and positioning of 36 experimental birds was registered. If birds were found in the box containing high density, low density, or in the central region, they received a 1, -1, or a 0 score, respectively. Birds with final summed scores of ≥3 or ≤-3 were categorized as HD or LD, respectively. Same category groups (HD or LD) were evaluated in their home box (undisturbed) and in a resident/intruder test when 38 d old. A higher proportion of LD than HD groups (5/6 vs. 1/6, respectively) showed at least one aggressive pecking event during a 1-h trial. The LD groups also showed a higher number of aggressive pecking events than HP groups. When an unfamiliar intruder (either HD or LD) was incorporated during 5 min in the HD or LD box, LD resident quail showed shorter latencies and a higher number of aggressive pecking events toward the intruder bird than their HD counterparts. The early individual permanence in the DRP test could be considered a consequence of a different adaptability strategy for group living. This novel test could be relevant for selection programs aiming to obtain birds better suited for rearing in high-density conditions.

Background: The strategy of salvage transplantation for patients with hepatocellular carcinoma is based on the premise that tumour recurrence will be still transplantable at the time of recurrence. However, patients can not only present non-transplantable recurrence but can also be over the age limit accepted for transplantation., Aims: To measure the risk of being too old for salvage transplantation of patients resected for hepatocellular carcinoma within Milan criteria., Methods: A Markov simulation model was developed on the basis of published literature., Results: The risk of being too old for salvage transplantation depends on the time-span between age at hepatic resection and age limit, and the expected median waiting-time. Patients resected at an age 2 or 3 years below the age limit carry a risk of being too old that overcomes the probability of receiving transplantation. Salvage strategy can cause harm that depends on the tumour characteristics and degree of portal hypertension, becoming maximal for patients with multiple tumours, clinical signs of portal hypertension and increased bilirubin levels., Conclusions: The best strategy to adopt should be balanced between the risk of being too old and the expected transplant benefit, but salvage strategy could be pursued if it did not turn into significant harm in comparison to primary transplantation., (Copyright © 2012 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.)

Malignant pleural mesothelioma (MPM), a cancer of the serosal pleural cavities, is one of the most aggressive human tumors. In order to identify genes crucial for the onset and progression of MPM, we performed an extensive literature review focused on transcriptome studies (RTS). In this kind of studies a great number of transcripts are analyzed without formulating any a priori hypothesis, thus preventing any bias coming from previously established knowledge that could lead to an over-representation of specific genes. Each study was thoroughly analyzed paying particular attention to: (i) the employed microarray platform, (ii) the number and type of samples, (iii) the fold-change, and (iv) the statistical significance of deregulated genes. We also performed data mining (DM) on MPM using three different tools (Coremine, SNPs3D, and GeneProspector). Results from RTS and DM were compared in order to restrict the number of genes potentially deregulated in MPM. Our main requirement for a gene to be a "mesothelioma gene" (MG) is to be reproducibly deregulated among independent studies and confirmed by DM. A list of MGs was thus produced, including PTGS2, BIRC5, ASS1, JUNB, MCM2, AURKA, FGF2, MKI67, CAV1, SFRP1, CCNB1, CDK4, and MSLN that might represent potential novel biomarkers or therapeutic targets for MPM. Moreover, it was found a sub-group of MGs including ASS1, JUNB, PTGS2, EEF2, SULF1, TOP2A, AURKA, BIRC5, CAV1, IFITM1, PCNA, and PKM2 that could explain, at least in part, the mechanisms of resistance to cisplatin, one first-line chemotherapeutic drug used for the disease. Finally, the pathway analysis showed that co-regulation networks related to the cross-talk between MPM and its micro-environment, in particular involving the adhesion molecules, integrins, and cytokines, might have an important role in MPM. Future studies are warranted to better characterize the role played by these genes in MPM., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Ligand-specific negative regulation of cytokine-induced signaling relies on down regulation of the cytokine receptors. Down regulation of the IFNAR1 sub-unit of the Type I interferon (IFN) receptor proceeds via lysosomal receptor proteolysis, which is triggered by ubiquitination that depends on IFNAR1 serine phosphorylation. While IFN-inducible phosphorylation, ubiquitination, and degradation requires the catalytic activity of the Tyk2 Janus kinase, here we found the ligand- and Tyk2-independent pathway that promotes IFNAR1 phosphorylation, ubiquitination, and degradation when IFNAR1 is expressed at high levels. A major cellular kinase activity that is responsible for IFNAR1 phosphorylation in vitro does not depend on either ligand or Tyk2 activity. Inhibition of ligand-independent IFNAR1 degradation suppresses cell proliferation. We discuss the signaling events that might lead to ubiquitination and degradation of IFNAR1 via ligand-dependent and independent pathways and their potential physiologic significance.

The broad and often contrasting effects of type I interferons (IFNs) in innate and adaptive immunity are belied by the signaling via a single receptor, IFN-alpha receptor (IFNAR). Here, we show that IFN-alpha/beta induces opposing effects on the immunologic outcome of antigen cross-presentation depending on dendritic cell (DC) maturation status. Despite equivalent IFNAR expression, immature conventional DCs (cDCs) activate STAT1 in response to IFN-alpha/beta, whereas exposure of mature DCs to IFN-alpha/beta results in signaling via STAT4. Microarray analysis revealed numerous transcriptional changes resulting from the altered signaling. Importantly, STAT1 signaling resulted in significant inhibition of CD40L-induced IL-12 production, accounting for the inhibition of CD8+ T-cell activation. These data provide evidence for a molecular switch in signaling pathways concomitant with DC maturation that offers a novel mechanism by which DCs modulate the integration of signals from the surrounding environment.

Objective: To determine the occurrence of carpal tunnel syndrome (CTS) and ulnar neuropathy at the elbow (UNE) in a cohort of floor cleaners and to check differences between workers with and without CTS., Methods: All female floor cleaners of three major hospitals in Tuscany (Italy) were contacted. Clinical and electrophysiological severity of CTS and UNE were evaluated with standardized scales and symptoms were assessed with the self-administered Boston Questionnaire (BQ); demographic and non-occupational factors and durations of current and previous occupations were recorded. Univariate analysis of risk factors was performed in workers with and without CTS. Logistic regression was used to evaluate the capacity of independent variables to predict CTS., Results: Out of a total of 179 cleaners, 145 (81%)-mean age 39.6 years (20-64 years)-were enrolled in the study; 70 (48%) had CTS (diagnosis based on clinical and electrophysiological findings). BQ symptom and hand function scores were anomalous in 108 (74%) and 84 (58%) subjects, respectively. UNE was detected in 7/103 women. Univariate analysis showed that cleaners with CTS were older, had greater BMI and longer exposure to cleaning with previous employers than those without CTS. In the logistic regression, the only predictor of CTS was cleaning with previous employers (O.R. 12.1, 95% CI 3-49.9)., Conclusions: These results indicate a high occurrence of CTS in floor cleaners; UNE is less frequent than CTS, presumably due to repetitive movements that stress wrists more than elbows. The only predictive factor of CTS was cleaning as an occupation with previous employers. Therefore, the actual risk factor for CTS could not be cleaning per se, but how this job is performed.

Interleukin-12 (IL-12) is a cytokine that plays a central role in the control of cell-mediated immunity. We have previously shown that transforming growth factor-beta1 (TGF-beta) inhibitory effects on human primary allogeneic cytotoxicity and proliferative responses interfere with IL-12 pathway. The present study was undertaken to further elucidate the biochemical basis of the functional interaction between these two cytokines and to define the site of TGF-beta action on the signaling pathway activated by IL-12. Our data indicate that TGF-beta induced an inhibition of interferon-gamma (IFN-gamma) production without affecting the IL-12Rbeta1 and IL-12Rbeta2 subunits mRNA expression by activated T cells. We further show that TGF-beta has a significant inhibitory effect on the early signal transduction events following interaction of IL-12 with its receptor on activated T cells, resulting in the inhibition of both JAK2 and Tyk2 phosphorylation. In addition, TGF-beta was found to significantly inhibit IL-12-induced phosphorylation of the STAT4 transcription factor. Electrophoretic mobility shift assay indicated that TGF-beta induced a decrease in IL-12-induced STAT4 DNA binding activity in T lymphocytes. This study suggests that TGF-beta influences IL-12 responsiveness at least in part by inhibiting early signaling events essential to gene induction in IL-12-activated T cells.

Various methods are now available to identify the molecular partners of the component of a signal transduction pathway. Some interactions, however, can be technically difficult to detect because they depend upon transient tyrosine phosphorylation. Here, we present a simple affinity chromatography approach based on synthetic phosphopeptides to purify potential partners of phosphotyrosine-containing proteins. With this approach, we confirm the previously characterized interaction between Grb2 and the EGF receptor, and we identify novel partners of the IGF-1 receptor and of the JAK proteins. Methenyltetrahydrofolate synthetase (MTHFS) was identified as a potential mediator of IGF-1R dependent transformation. P85alpha, the regulatory subunit of PI3 kinase, was identified as one of four proteins recruited by a phosphopeptide mimicking a motif conserved in all JAK family members.

A single strand conformation polymorphism analysis on polymerase chain reaction (PCR)-amplified genomic DNA was used for the detection of DNA mutations in acid formalin-fixed, paraffin-embedded tissues. Fifty non-small cell lung cancers were screened for mutations in the exon 5 of the p53 tumor suppressor gene. Structural abnormalities of the gene were found in nine (18%) of the tumors examined. The results show that paraffin-embedded tissues fixed in unbuffered formalin can be a good source of DNA for a mobility shift analysis by neutral polyacrylamide gel electrophoresis. The described procedure allows retrospective genetic studies on paraffin blocks available in all pathology departments.

Objective: Our purpose was to investigate the efficacy of postsurgical treatment with nafarelin in women with advanced endometriosis., Study Design: Eligible for trial were women < or = 38 years old with unexplained infertility with or without chronic pelvic pain and stage III or IV endometriosis according to the American Fertility Society, revised, classification who underwent laparotomy as first surgical treatment for debulking or radical surgery of endometriotic lesions. Patients were assigned according to a randomization list to nasal nafarelin, 400 micrograms/day (36 subjects) or placebo nasal spray (39 subjects) for 3 months. Pelvic pain was assessed before first surgery and at the 12-month follow-up visit in women with pelvic pain by means of a multidimensional score system and a 10-point linear pain scale., Results: No marked differences in pain scores emerged among women allocated to different treatments. The mean reduction of the multidimensional score was 3.6 and 4.0, respectively, in women allocated to nafarelin and placebo and of the 10-point linear scale scores was 7.0 and 6.9. These differences were not statistically significant. Within 1 year from randomization, of the 36 women allocated to nafarelin and the 39 allocated to placebo, seven (19%) and seven (18%), respectively, became pregnant., Conclusion: This study suggests that medical treatment with nafarelin does not markedly improve pelvic pain and short-term reproductive prognosis in women with stages III and IV endometriosis.