Analysis of human MDM4 variants in papillary thyroid carcinomas reveals new potential markers of cancer properties

A wild-type (wt) p53 gene characterizes thyroid tumors, except for the rare anaplastic histotype. Because p53 inactivation is a prerequisite for tumor development, alterations of p53 regulators represent an alternative way to impair p53 function. Indeed, murine double minute 2 (MDM2), the main p53 negative regulator, is overexpressed in many tumor histotypes including those of the thyroid. A new p53 regulator, MDM4 (a.k.a. MDMX or HDMX) an analog of MDM2, represents a new oncogene although its impact on tumor properties remains largely unexplored. We estimated levels of MDM2, MDM4, and its variants, MDM4-S (originally HDMX-S) and MDM4-211 (originally HDMX211), in a group of 57 papillary thyroid carcinomas (PTC), characterized by wt tumor protein 53, in comparison to matched contra-lateral lobe normal tissue. Further, we evaluated the association between expression levels of these genes and the histopathological features of tumors. Quantitative real-time polymerase chain reaction revealed a highly significant downregulation of MDM4 mRNA in tumor tissue compared to control tissue (P < 0.0001), a finding confirmed by western blot on a subset of 20 tissue pairs. Moreover, the tumor-to-normal ratio of MDM4 levels for each individual was significantly lower in late tumor stages, suggesting a specific downregulation of MDM4 expression with tumor progression. In comparison, MDM2 messenger RNA (mRNA) and protein levels were frequently upregulated with no correlation with MDM4 levels. Lastly, we frequently detected overexpression of MDM4-S mRNA and presence of the aberrant form, MDM4-211 in this tumor group. These findings indicate that MDM4 alterations are a frequent event in PTC. It is worthy to note that the significant downregulation of full-length MDM4 in PTC reveals a novel status of this factor in human cancer that counsels careful evaluation of its role in human tumorigenesis and of its potential as therapeutic target

Dual targeting of p53 and c-MYC selectively eliminates leukaemic stem cells

e Glasgow and Manchester Experimental Cancer Medicine Centres (ECMC), which are funded by CR-UK and the Chief Scientist’s Office (Scotland). We acknowledge the funders who have contributed to this work: MRC stratified medicine infrastructure award (A.D.W.), CR-UK C11074/A11008 (F.P., L.E.M.H., T.L.H., A.D.W.); LLR08071 (S.A.A., E.C.); LLR11017 (M.C.); SCD/04 (M.C.); LLR13035 (S.A.A., K.D., A.D.W., and A.P.); LLR14005 (M.T.S., D.V.); KKL690 (L.E.P.); KKL698 (P.B.); LLR08004 (A.D.W., A.P. and A.J.W.); MRC CiC (M.E.D.); The Howat Foundation (FACS support); Friends of Paul O’Gorman (K.D. and FACS support); ELF 67954 (S.A.A.); BSH start up fund (S.A.A.); MR/K014854/1 (K.D.)

Polycomb-Like 3 Promotes Polycomb Repressive Complex 2 Binding to CpG Islands and Embryonic Stem Cell Self-Renewal

Polycomb repressive complex 2 (PRC2) trimethylates lysine 27 of histone H3 (H3K27me3) to regulate gene expression during diverse biological transitions in development, embryonic stem cell (ESC) differentiation, and cancer. Here, we show that Polycomb-like 3 (Pcl3) is a component of PRC2 that promotes ESC self-renewal. Using mass spectrometry, we identified Pcl3 as a Suz12 binding partner and confirmed Pcl3 interactions with core PRC2 components by co-immunoprecipitation. Knockdown of Pcl3 in ESCs increases spontaneous differentiation, yet does not affect early differentiation decisions as assessed in teratomas and embryoid bodies, indicating that Pcl3 has a specific role in regulating ESC self-renewal. Consistent with Pcl3 promoting PRC2 function, decreasing Pcl3 levels reduces H3K27me3 levels while overexpressing Pcl3 increases H3K27me3 levels. Furthermore, chromatin immunoprecipitation and sequencing (ChIP-seq) reveal that Pcl3 co-localizes with PRC2 core component, Suz12, and depletion of Pcl3 decreases Suz12 binding at over 60% of PRC2 targets. Mutation of conserved residues within the Pcl3 Tudor domain, a domain implicated in recognizing methylated histones, compromises H3K27me3 formation, suggesting that the Tudor domain of Pcl3 is essential for function. We also show that Pcl3 and its paralog, Pcl2, exist in different PRC2 complexes but bind many of the same PRC2 targets, particularly CpG islands regulated by Pcl3. Thus, Pcl3 is a component of PRC2 critical for ESC self-renewal, histone methylation, and recruitment of PRC2 to a subset of its genomic sites

Lamotrigine Subacute Cutaneous Lupus Erythematous

Objective: Lamotrigine is a new generation anticonvulsant, also used in bipolar type 1 disorders to prevent occurrence of mood episodes not responding to traditional antidepressant, which is an increasing indication worldwide. Skin adverse events occur in 5-10% of patients, but are usually mild and self-limiting. The occurrence of drug-induced lupus erythematosus is an uncommon, but severe condition, usually requiring discontinuation of the drug. Methods: A spontaneous pharmacovigilance case reporting is presented, following the causality assessment criteria, including temporal association with drug exposure, exclusion of other etiologies, dechallenge and rechallenge. Skin biopsy for normal histopathology and direct immunofluorescence was performed to assess the diagnosis of lupus erythematosus. Results: A 77 years old woman was hospitalized in the Dermatology Clinic of Cagliari University for a lamotrigine Induced Subacute Cutaneous Lupus Erythematosus, confirmed by improvement after drug dismission, and immediate recurrence at re-exposure. The last measure was performed on request of the neurologist with the consent of the patient, because of a high suicidal risk, not responding to traditional antidepressants, and well controlled by lamotrigine. Another peculiarity of our observation is a potential dose-related effects, as the patients was assuming lamotrigine from 2 years, and skin manifestations actually began 3 weeks after the dosage was doubled to 100 mg daily. Conclusion: If confirmed by other observation, drug-induced subacute cutaneous lupus erythematosus might be dose-dependent, and careful clinician’s dosage monitoring might prevent occurrence, preserving the lamotrigine use for the many patients who need it. Dermatologist referral is warrant in front of any suspect cutaneous adverse reaction due to psychotropic medications, to provide prompt recognition, adequate assessment and supportive management of such difficult and delicate patients

COVID-19 pulmonary infection in erythrodermic psoriatic patient with oligodendroglioma: safety and compatibility of apremilast with critical intensive care management

Novel coronavirus 2019 (SARS-CoV2) pandemic has particularly affected Italy, with a profound impact on the therapeutic strategy for complex disorder such as psoriasis, whose extensive skin damage might expose to an increased infective risk compared to the general population. Psoriasis treatment relies on immunosuppression, and although most experts agree that the benefit-to risk-ratio is in favor of maintaining selective biologic therapies, and small molecules such as apremilast, they recommend dismission if severe COVID-19 symptoms occur

Knock-down of Kaiso induces proliferation and blocks granulocytic differentiation in blast crisis of chronic myeloid leukemia

Abstract Background Kaiso protein has been identified as a new member of the POZ-ZF subfamily of transcription factors that are involved in development and cancer. There is consistent evidence of the role of Kaiso and its involvement in human tumorigenesis but there is no evidence about its role in hematopoietic differentiation or establishment of chronic myeloid leukemia (CML). We used, normal K562 cell line, established from a CML patient in blast crisis, and imatinib-resistant K562 cell line, to investigate the specific distribution of Kaiso and their contribution to the cell differentiation status of the blast crisis of CML (CML-BP). Results We found cytoplasmic expression of Kaiso, in K562 cells and patients, confirmed by immunofluorescence, immunohistochemistry and western blot of cytoplasmic protein fraction. Kaiso was weakly expressed in the imatinib-resistant K562 cell line confirmed by immunofluorescence and western blot. The cytoplasmic expression of Kaiso was not modified when the K562 cells were treated for 16 h with imatinib 0.1 and 1 μM. In our study, small interfering RNA (siRNA) was introduced to down regulate the expression of Kaiso and p120ctn in K562 cell line. Kaiso and p120ctn were down regulated individually (siRNA-Kaiso or siRNA-p120ctn) or in combination using a simultaneous co-transfection (siRNA-Kaiso/p120ctn). We next investigated whether knockdown either Kaiso or p120ctn alone or in combination affects the cell differentiation status in K562 cells. After down regulation we analyzed the expression of hematopoietic cell differentiation and proliferation genes: SCF, PU-1, c-MyB, C/EBPα, Gata-2 and maturation markers of hematopoietic cells expressed in the plasma membrane: CD15, CD11b, CD33, CD117. The levels of SCF and c-MyB were increased by 1000% and 65% respectively and PU-1, Gata-2 and C/EBPα were decreased by 66%, 50% and 80% respectively, when Kaiso levels were down regulated by siRNA. The results were similar when both Kaiso and p120ctn were down regulated by siRNA. The increased expression of SCF and decreased expression of GATA-2 could be responsible by the higher cell viability detected in K562 cells double knock-down of both Kaiso and p120ctn. Finally, we studied the effect of knock-down either Kaiso or p120ctn, alone or in combination on CD15, CD11b, CD33 and Cd117 expression. Using siRNA approach a reduction of 35%, 8% and 13% in CD15, CD33 and CD117 levels respectively, were achieved in all transfections, when compared to scrambled knock-down cells. Conclusion These results suggest that both Kaiso and p120ctn, contributes to maintaining the differentiated state of the K562 cells and similar to other cancers, cytoplasmic localization of Kaiso is related to a poor prognosis in CML-BP. By the broad and profound effects on the expression of genes and markers of hematopoietic differentiation produced by Kaiso knock-down, these findings reveal Kaiso as a potential target for selective therapy of CML.</p

Protezione della vite dall'oidio con fungicidi impiegati a soglie crescenti di malattia

Nel biennio 2007-2008 \ue8 stata valutata l\u2019efficacia di metrafenone e kresoxim-methyl + boscalid nei confronti di Erysiphe necator, applicati a soglie crescenti di diffusione della malattia. Entrambi i fungicidi sono dotati di ottima efficacia iniziando la difesa alle soglie pi\uf9 basse di diffusione dell\u2019oidio, mentre quando applicati a partire dal 50-60% di diffusione non hanno contenuto l\u2019oidio in modo soddisfacente. Tuttavia, la miscela kresoximmethyl+ boscalid ha mostrato una capacit\ue0 bloccante superiore a metrafenone ed entrambi i fungicidi sono risultati pi\uf9 efficaci dei prodotti di riferimento, sebbene tali differenze non sempre sono risultate statisticamente significative. Aumenti del dosaggio di kresoximmethyl+ boscalid da 30 ml/hl a 40 ml/hl di formulato co

Isolation and characterization of bacteria associated with rice grain discolouration

In Italy, in spite the widespread use of fungicides to control blast and brown spot epidemics, the incidence of discoloured rice grains has recently increased. Widely grown rice varieties like \u2018Selenio\u2019, \u2018Loto\u2019, and \u2018Balilla\u2019 are highly susceptible to grain discolouration. Disease incidence is highest in early sown crops and it is negatively correlated to panicle brown spot incidence and severity. These observations together with lack of fungicides efficacy, suggest that bacteria may cause the disease, as it occurs in tropical countries. To have a better insight into grain discolouration aetiology in Italy we analysed the bacterial population associated with discoloured grains of cv Selenio (N=43) and other varieties (N=8). More than 400 bacterial isolates were obtained. DNA Repetitive Extragenic Palindromic (REP) sequences were amplified for each isolate, using the primer BOXA1R. More than 200 isolates had unique BOX-fingerprint and were further char-acterized with molecular and physiological analyses. About 80% of the bacterial population was composed by Gram-negative species: 60% of these isolates were fermenting, and the majority was Pantoea spp., while 40% resulted strictly aerobic. About 70 isolates were identified as Pseudomonas (sensu stricto) and cluster analysis of ITS1-RFLP data showed 9 major groups. Partial sequences of 16S rDNA of representative isolates were aligned and matched with the following species: Ps. fluorescens, Ps. stutzeri, Ps. fulva and Ps. putida. Acidovorax avenae sub. sp. avenae, a seed-transmitted and highly destructive bacterium in tropical countries, was found in 25% of the rice samples (N=21). Pathogenicity tests provided additional evidence that rice grain discolouration in Italy is due to bacteria