Management of early melanoma recurrence despite adjuvant anti-PD-1 antibody therapy☆.

BACKGROUND: Anti-programmed cell death protein 1 (PD-1) antibodies (PD1) prolong recurrence-free survival in high-risk resected melanoma; however, approximately 25%-30% of patients recur within 1 year. This study describes the pattern of recurrence, management and outcomes of patients who recur with adjuvant PD1 therapy. PATIENTS AND METHODS: Consecutive patients from 16 centres who recurred having received adjuvant PD1 therapy for resected stage III/IV melanoma were studied. Recurrence characteristics, management and outcomes were examined; patients with mucosal melanoma were analysed separately. RESULTS: Melanoma recurrence occurred in 147 (17%) of ∼850 patients treated with adjuvant PD1. In those with cutaneous melanoma (n = 136), median time to recurrence was 4.6 months (range 0.3-35.7); 104 (76%) recurred during (ON) adjuvant PD1 after a median 3.2 months and 32 (24%) following (OFF) treatment cessation after a median 12.5 months, including in 21 (15%) who ceased early for toxicity. Fifty-nine (43%) recurred with locoregional disease only and 77 (57%) with distant disease. Of those who recurred locally, 22/59 (37%) subsequently recurred distantly. Eighty-nine (65%) patients received systemic therapy after recurrence. Of those who recurred ON adjuvant PD1, none (0/6) responded to PD1 alone; 8/33 assessable patients (24%) responded to ipilimumab (alone or in combination with PD1) and 18/23 (78%) responded to BRAF/MEK inhibitors. Of those who recurred OFF adjuvant PD1, two out of five (40%) responded to PD1 monotherapy, two out of five (40%) responded to ipilimumab-based therapy and 9/10 (90%) responded to BRAF/MEK inhibitors. CONCLUSIONS: Most patients who recur early despite adjuvant PD1 develop distant metastases. In those who recur ON adjuvant PD1, there is minimal activity of further PD1 monotherapy, but ipilimumab (alone or in combination with PD1) and BRAF/MEK inhibitors have clinical utility. Retreatment with PD1 may have activity in select patients who recur OFF PD1.

Can platelet-rich plasma be an alternative to surgery for resistant chronic patellar tendinopathy in sportive people? Poor clinical results at 1-year follow-up.

INTRODUCTION AND PURPOSE: Patellar tendinopathy is a disease affecting particularly athletes. Platelet-rich plasma (PRP) injections have gained increasing interest for their potential benefits. Anyway, a tendon disease longer than 6 months should be considered as an indication for surgery. The aim of our study was to evaluate the efficacy of PRP in athletes with a severe chronic patellar tendinopathy longer than 6 months when surgery should be chosen. METHODS: We enrolled 17 sport practitioners (19 patellar tendons) who did not want to undergo surgery and who are nonresponders to other conservative treatments. We treated them with PRP and calculated the results using the visual analog scale (VAS), the Victorian Institute of Sport Assessment-Patellar (VISA-P) score, and Tegner Activity Scale. Every test has been conducted at T0, T1 (4 months), and T2 (12 months). RESULTS: We found a poor improvement at T1 and a clinical worsening at T2 through VAS. VISA-P showed a medium improvement both at T1 and T2. Tegner scale did not show improvements. CONCLUSIONS: Our study was not able to remove the doubts about the benefits of PRP in patellar tendinopathy, confirming ambiguous certainties. Further investigations are needed to assess its effectiveness.

Evaluating diagnostic accuracy of anti-tissue Transglutaminase IgA antibodies as first screening for Celiac Disease in very young children.

BACKGROUND: Small bowel biopsy is the gold standard for Celiac Disease (CD) diagnosis, nevertheless serum assays are the first step in ascertaining a diagnosis of CD. New ESPGHAN Criteria 2012 (European Society of Pediatric Gastroenterology Hepatology and Nutrition) suggest using exclusively anti-tissue Transglutaminase IgA antibodies (anti-tTGA) as initial approach to symptomatic subjects. The aim of our study was to evaluate the diagnostic accuracy of anti-tTGA as initial screening assay for CD in a large cohort of pediatric patients. METHODS: We selected 730 subjects aged between 6 months and 4 years ("Group A") and 348 subjects younger than 2 years (which are part of the 730 subjects) ("Group B"). We performed anti-Deamidated Gliadin Peptides IgA and IgG antibodies (a-DGP IgA/IgG) and anti-tTGA assays by ELISA test. We evaluated the agreement between anti-tTGA and a-DGP IgA/IgG assays and compared the diagnostic accuracy of a-DGP IgA/IgG with that of anti-tTGA in both groups of patients. RESULTS: There was a substantial agreement between anti-tTGA and a-DGP IgA in "Group A" and an almost perfect agreement in "Group B"; the strength of agreement between anti-tTGA and a-DGP IgG was moderate in "Group A" and substantial in "Group B". anti-tTGA were more sensitive and specific than a-DGP IgA/IgG in both groups. CONCLUSIONS: anti-tTGA could be used as initial screening assay for CD in all subjects from 6 months of age according to ESPGHAN Criteria 2012.

LAMC2 enhances the metastatic potential of lung adenocarcinoma.

Lung cancer is the number one cancer killer, and metastasis is the main cause of high mortality in lung cancer patients. However, mechanisms underlying the development of lung cancer metastasis remain unknown. Using genome-wide transcriptional analysis in an experimental metastasis model, we identified laminin γ2 (LAMC2), an epithelial basement membrane protein, to be significantly upregulated in lung adenocarcinoma metastatic cells. Elevated LAMC2 increased traction force, migration, and invasion of lung adenocarcinoma cells accompanied by the induction of epithelial-mesenchymal transition (EMT). LAMC2 knockdown decreased traction force, migration, and invasion accompanied by EMT reduction in vitro, and attenuated metastasis in mice. LAMC2 promoted migration and invasion via EMT that was integrin ß1- and ZEB1-dependent. High LAMC2 was significantly correlated with the mesenchymal marker vimentin expression in lung adenocarcinomas, and with higher risk of recurrence or death in patients with lung adenocarcinoma. We suggest that LAMC2 promotes metastasis in lung adenocarcinoma via EMT and may be a potential therapeutic target.

Complicated diverticulitis in kidney transplanted patients: analysis of 717 cases.

INTRODUCTION: This study aims to investigate possible risk factors for diverticulitis in kidney transplant recipients affected by colonic diverticulosis. METHODS AND RESULTS: We investigated 717 patients transplanted between 2000 and 2010. Diverticular disease was endoscopically diagnosed in 17 of 717 examined patients. Eight patients were diagnosed with autosomal dominant polycystic kidney disease (ADPKD); 9 of 17 patients underwent emergency surgery. We performed Hartmann's procedure on all patients, with a second stage performed at least 6 months later. DISCUSSION: Although the incidence of colonic diverticular perforation in kidney transplanted patients is similar to that observed in the general population, perforation in immunosuppressed patients is associated with a higher morbidity/mortality rate. In our study, the incidence of perforation is 1.25% (9 of 717), with almost half of the cases observed in patients with ADPKD (4 of 9). Such an observation is consistent with published data, in which patients with ADPKD are reported to more frequently develop colonic diverticulosis and its complications. One possible explanation might be related to a belated diagnosis of diverticulitis, which could initially simulate an inflammatory disease as a consequence of renal cysts. Also, steroids seem to be a predisposing factor for colonic perforation in these patients. CONCLUSIONS: A timely surgery can significantly reduce mortality. In cases of elective surgery, mortality and morbidity are similar to those of immunocompetent patients; accordingly, this is the goal to be pursued. Early signs and symptoms are often masked by immunosuppressive therapy. In these patients, surgeons should always perform (1) abdominal computed tomography scanning and, in the presence of diverticulitis, reduce or withdraw immunosuppressive therapy; and (2) early surgery, with Hartmann's procedure being, in our opinion, the best choice. Before transplantation, elective surgery for colonic resection should be considered in patients with ADPKD or with a history of 1 or more episodes of acute diverticulitis who then regressed with medical therapy.

First Report of Fusarium oxysporum f.sp. pisi as Causal Agent of Root and Crown Rot on Chickpea (Cicer arietinum) in Southern Italy.

Epidemiological investigations in representative chickpea (Cicer arietinum L.) fields in southern Italy (Larino, Campobasso, 41°50'45″ N, 14°55'28″ E) identified severe withering (25 to 51%) of plants during flowering. Diseased plants showed a reduced total root biomass associated with less vigorous and chlorotic foliage. Browning and necrosis of subcortical and xylematic tissues of the crown and main roots were observed in affected plants. Symptomatic root and stem portions from 50 plants were sampled, surface disinfected with a sodium hypochlorite water solution (2% v/v for 2 min), rinsed with sterile distilled water, and placed in petri dishes containing potato dextrose agar with streptomycin sulfate (200 mg/l) and incubated at 25°C for 10 days. The most frequent fungal colony isolated showed macro- and microscopic characters specific of the genus Fusarium (3), with falcate and three-septate macroconidia (24.0 to 43.8 µm long) and microconidia (6.8 to 10.4 µm long) with zero or one septa. The ribosomal DNA of the fungal isolate processed by PCR using the ITS1F/ITS4 primers (2) produced an amplicon of 545 bp (ENA, Accession No. HG423346). A BLAST search with the amplified sequence in the database of the International Mycological Association ( www.mycobank.org ) revealed 99% identity with F. oxysporum sequences. Additional molecular analysis using the specific primers Foc0-12/Foc0-12rf for F. oxysporum f.sp. ciceris (Foc) produced an amplicon only in the chickpea virulent strain Foc-7952, race 0 (1) used as control; furthermore, PCR amplification for the Pisatin Demetylase gene by using the specific primers PDAF2a and PDAR3a (4) yielded the expected amplicon only for the new isolate, whereas no amplification was obtained with the control strain Foc-7952. Pathogenicity assays were carried out to complete Koch's postulates. To this aim, horticultural peat was infested with a conidial suspension (1 × 104 conidia/g of soil) from the new fungal pathogen, dispensed in plastic pots, and sown with surface sterilized seeds of chickpea (cv. Real, ISEA, Italy). Uncontaminated peat was used as control. For both treatments, 3 replicates of 10 seeds were used and experiments repeated twice. The plastic pots were kept in a growth chamber (28°C; 70% RH; 15/9 h light/dark) where the first disease symptoms on plants appeared 20 days after sowing. At the end of the experiments, all plants inoculated with the new isolate showed a high disease severity (98%), whereas non-inoculated plants remained healthy. The seedlings from infested soil demonstrated the same symptoms previously observed in the field, and after re-isolation, the causal agent demonstrated the same morphological features of the isolate used for inoculation. Pathogenicity tests were performed on pea, faba bean, melon, and tomato by using three cultivars for each crop. The results demonstrated high virulence of the new isolate of F. oxysporum f.sp. pisi (Fop) on both chickpea and pea with seed germination reduction, rot on main and secondary roots and cotyledonary leaves, and root biomass reduction and foliage chlorosis. No symptoms were observed on other inoculated vegetal species. Collectively, data of our investigation allow us to affirm that this is the first report of Fop as a new pathogen of chickpea. This result has great economic importance since it enables specific monitoring and management plans for this new disease caused by Fop on chickpea, a key crop for human and animal nutrition. References: (1) M. M. Jiménez-Gasco and R. M. Jiménez-Díaz, Phytopathology 93:201, 2003. (2) I. Larena et al. J. Biotechnol. 75:187, 1999. (3) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (4) N. A. Milani et al. Fungal Genet. Biol. 933:942, 2012.

PATZ1 interacts with p53 and regulates expression of p53-target genes enhancing apoptosis or cell survival based on the cellular context.

PATZ1 is a transcriptional factor functioning either as an activator or a repressor of gene transcription depending upon the cellular context. It appears to have a dual oncogenic/anti-oncogenic activity. Indeed, it is overexpressed in colon carcinomas, and its silencing inhibits colon cancer cell proliferation or increases sensitivity to apoptotic stimuli of glioma cells, suggesting an oncogenic role. Conversely, the development of B-cell lymphomas, sarcomas, hepatocellular carcinomas and lung adenomas in Patz1-knockout (ko) mice supports its tumour suppressor function. PATZ1 role in mouse lymphomagenesis is mainly because of the involvement of PATZ1 in BCL6-negative autoregulation. However, this does not exclude that PATZ1 may be involved in tumorigenesis by other mechanisms. Here, we report that PATZ1 interacts with the tumour suppressor p53 and binds p53-dependent gene promoters, including those of BAX, CDKN1A and MDM2. Knockdown of PATZ1 in HEK293 cells reduces promoter activity of these genes and inhibits their expression, suggesting a role of PATZ in enhancing p53 transcriptional activity. Consistently, Patz1-ko mouse embryonic fibroblasts (MEFs) show decreased expression of Bax, Cdkn1a and Mdm2 compared with wild-type (wt) MEFs. Moreover, Patz1-ko MEFs show a decreased percentage of apoptotic cells, either spontaneous or induced by treatment with 5-fluorouracil (5FU), compared with wt controls, suggesting a pro-apoptotic role for PATZ1 in these cells. However, PATZ1 binds p53-target genes also independently from p53, exerting, in the absence of p53, an opposite function on their expression. Indeed, knockdown of PATZ1 in p53-null osteosarcoma cells upregulates BAX expression and decreases survival of 5FU-treated cells, then suggesting an anti-apoptotic role of PATZ1 in p53-null cancer cells. Therefore, these data support a PATZ1 tumour-suppressive function based on its ability to enhance p53-dependent transcription and apoptosis. Conversely, its opposite and anti-apoptotic role in p53-null cancer cells provides the perspective of PATZ1 silencing as a possible adjuvant in the treatment of p53-null cancer.

miR-130a targets MET and induces TRAIL-sensitivity in NSCLC by downregulating miR-221 and 222.

Non-small cell lung cancer (NSCLC) accounts for ∼80% of all lung cancers. Although some advances in lung cancer therapy have been made, patient survival is still quite poor. Two microRNAs, miR-221 and miR-222, upregulated by the MET proto-oncogene, have been already described to enhance cell survival and to induce TNF-related apoptosis-inducing ligand (TRAIL) resistance in NSCLC cell lines, through the downregulation of p27(kip1), PTEN and TIMP3. Here, we further investigated this pathway and showed that miR-130a, expressed at low level in lung cancer cell lines, by targeting MET was able to reduce TRAIL resistance in NSCLC cells through the c-Jun-mediated downregulation of miR-221 and miR-222. Moreover, we found that miR-130a reduced migratory capacity of NSCLC. A better understanding of MET-miR-221 and 222 axis regulation in drug resistance is the key in developing new strategies in NSCLC therapy.

Downregulation of HMGA-targeting microRNAs has a critical role in human pituitary tumorigenesis.

Previous studies have demonstrated that high mobility group A proteins have a critical role on the onset of human pituitary adenomas. Indeed, both high mobility group A (HMGA) genes are overexpressed in pituitary adenomas, and consistently transgenic mice overexpressing either the Hmga1 or the Hmga2 gene develop mixed growth hormone/prolactin (GH-PRL)-secreting pituitary adenomas. Trisomy of chromosome 12, where HMGA2 is located, and/or amplification of the HMGA2 gene locus account for the HMGA2 overexpression in most human prolactinomas. Conversely, HMGA1 overexpression is not associated to any rearrangement or amplification of the HMGA1 locus. We have first identified micro RNAs (miRNAs) able to target both HMGA1 and HMGA2 messenger RNAs. Then, all of these miRNAs have been found downregulated in pituitary adenomas of different histotypes, compared with normal pituitary. Interestingly, their downregulation was also observed in nonfunctioning pituitary adenomas where HMGA2 overexpression is not associated to any alteration of the HMGA2 locus. Functional studies show that all these HMGA-targeting miRNAs inhibit the proliferation of the rat pituitary adenoma cell line GH3. Therefore, these results indicate that the downregulation of the miRNAs able to target the HMGA genes could contribute to increase HMGA protein levels in human pituitary adenomas, and then to pituitary tumorigenesis.

Rapidly controlled outbreak of Serratia marcescens infection/colonisations in a neonatal intensive care unit, Pescara General Hospital, Pescara, Italy, April 2011.

In April 2011, an outbreak of Serratia marcescens infection/ colonisations occurred in the neonatal intensive care unit of Pescara General Hospital. Rapid microbiological investigations lead to identification of five cases of likely cross-transmission from a neonate hospitalised for S. marcescens sepsis: four infections and one neonate colonised post-mortem. Two low birth weight neonates died. The environmental investigation detected S. marcescens from two soap dispensers. Strict hygiene measures lead to early interruption of the outbreak, without recurrences to date.

MR in the evaluation of new anterior cruciate ligament and tibial tunnel position: correlation with clinical and functional features.

PURPOSE: This study aimed to evaluate correlations between the position of the tibial tunnel, its alignment with the ligament-screw system, presence of intratunnel fluid, position of the tibial tunnel with respect to the Blumensaat line and clinical knee stability in patients who underwent arthroscopic reconstruction of the anterior cruciate ligament (ACL), by using magnetic resonance (MR) imaging. MATERIALS AND METHODS: Forty-eight patients (40 men, eight women; mean age, 31 years) underwent arthroscopic reconstruction of the ACL using double-strand semitendinosus and gracilis tendons. The new ACL was fixed to the tibial tunnel using Bio-Intrafix (Mitek). All patients underwent MR imaging 12 months after surgery and clinical evaluation at 6 and 12 months using the International Knee Documentation Committee (IKDC) scoring system. MR imaging and clinical features were correlated using the Mann-Whitney U test for continuous variables and Fisher's exact test for categorical variables. RESULTS: Forty-one patients were clinically stable (groups A and B according to the IKDC test) and seven were unstable (group C). Mean values of tibial tunnel position in clinically unstable vs stable patients were, respectively, -3.6 ±3.8 mm vs. -2.8±3.8 mm in relation to the Blumensaat line (p=0.5712) and 77.3°±11.3 vs. 72.5°±5.5 as concerned the angle measured on the coronal view of the new ACL (p=0.3248); fluid was present in the tibial tunnel in 42.9% and 9.8% of cases, respectively (p=0.2104). MR imaging showed misalignment of ligament screw and tibial tunnel in 57.1% of patients in group C and in 12.2% in groups A and B (p=0.017). CONCLUSIONS: Misalignment of the ligament-screw system and the tibial tunnel and the presence of fluid in the tibial tunnel appear to be directly correlated with clinical instability.

HMGA proteins promote ATM expression and enhance cancer cell resistance to genotoxic agents.

DNA-damaging therapies represent a keystone in cancer treatment. Unfortunately, many tumors often relapse because of a group of cancer cells, which are resistant to conventional therapies. High-mobility group A (HMGA) proteins has a key role in cell transformation, and their overexpression is a common feature of human malignant neoplasias, representing a poor prognostic index often correlated to anti-cancer drug resistance. Our previous results demonstrated that HMGA1 is a substrate of ataxia-telangiectasia mutated (ATM), the main cellular sensor of genotoxic stress. Here we also report thatHMGA2, the other member of the HMGA family, is a novel substrate of ATM. Interestingly, we found that HMGA proteins positively regulate ATM gene expression. Moreover, induction of ATM kinase activity by DNA-damaging agents enhances HMGA-dependent transcriptional activation of ATM promoter, suggesting that ATM expression is modulated by a DNA-damage- and HMGA-dependent positive feedback loop. Finally, inhibition of HMGA expression in mouse embryonic fibroblasts and in cancer cells strongly reduces ATM protein levels, impairing the cellular DNA-damage response and enhancing the sensitivity to DNA-damaging agents. These findings indicate this novel HMGA-ATM pathway as a new potential target to improve the effectiveness of conventional anti-neoplastic treatments on the genotoxic-drug resistant cancer cells.

Osteoblasts extracellular matrix induces vessel like structures through glycosylated collagen I.

Extracellular matrix (ECM) plays a fundamental role in angiogenesis affecting endothelial cells proliferation, migration and differentiation. Vessels-like network formation in vitro is a reliable test to study the inductive effects of ECM on angiogenesis. Here we utilized matrix deposed by osteoblasts as substrate where the molecular and structural complexity of the endogenous ECM is preserved, to test if it induces vessel-like network formation by endothelial cells in vitro. ECM is more similar to the physiological substrate in vivo than other substrates previously utilized for these studies in vitro. Osteogenic ECM, prepared in vitro from mature osteoblasts at the phase of maximal deposition and glycosylation of collagen I, induces EAhy926, HUVEC, and HDMEC endothelial cells to form vessels-like structures and promotes the activation of metalloproteinase-2 (MMP-2); the functionality of the p-38/MAPK signaling pathway is required. Osteogenic ECM also induces a transient increase of CXCL12 and a decrease of the receptor CXCR4. The induction of vessel-like networks is dependent from proper glycosylation of collagens and does not occur on osteogenic ECMs if deglycosylated by -galactosidase or on less glycosylated ECMs derived from preosteoblasts and normal fibroblasts, while is sustained on ECM from osteogenesis imperfecta fibroblasts only when their mutation is associated with over-glycosylation of collagen type I. These data support that post-translational glycosylation has a role in the induction in endothelial cells in vitro of molecules conductive to self-organization in vessels-like structures.

Capillary filling in microchannels with wall corrugations: a comparative study of the Concus-Finn criterion by continuum, kinetic, and atomistic approaches.

We study the impact of wall corrugations in microchannels on the process of capillary filling by means of three broadly used methods: computational fluid dynamics (CFD), lattice Boltzmann equations (LBE), and molecular dynamics (MD). The numerical results of these approaches are compared and tested against the Concus-Finn (CF) criterion, which predicts pinning of the contact line at rectangular ridges perpendicular to flow for contact angles of theta > 45 degrees . Whereas for theta = 30, 40 (no flow), and 60 degrees (flow) all methods are found to produce data consistent with the CF criterion, at theta = 50 degrees the numerical experiments provide different results. Whereas the pinning of the liquid front is observed both in the LB and CFD simulations, MD simulations show that molecular fluctuations allow front propagation even above the critical value predicted by the deterministic CF criterion, thereby introducing a sensitivity to the obstacle height.

Regulation of microRNA expression by HMGA1 proteins.

The High Mobility Group proteins HMGA1 are nuclear architectural factors that play a critical role in a wide range of biological processes. Since recent studies have identified the microRNAs (miRNAs) as important regulators of gene expression, modulating critical cellular functions such as proliferation, apoptosis and differentiation, the aim of our work was to identify the miRNAs that are physiologically regulated by HMGA1 proteins. To this purpose, we have analysed the miRNA expression profile of mouse embryonic fibroblasts (MEFs) carrying two, one or no Hmga1 functional alleles using a microarray (miRNA microarray). By this approach, we found a miRNA expression profile that differentiates Hmga1-null MEFs from the wild-type ones. In particular, a significant decrease in miR-196a-2, miR-101b, miR-331 and miR-29a was detected in homozygous Hmga1-knockout MEFs in comparison with wild-type cells. Consistently, these miRNAs are downregulated in most of the analysed tissues of Hmga1-null mice in comparison with the wild-type mice. ChIP assay shows that HMGA1 is able to bind regions upstream of these miRNAs. Moreover, we identified the HMGA2 gene product as a putative target of miR-196a-2, suggesting that HMGA1 proteins are able to downregulate the expression of the other member of the HMGA family through the regulation of the miR-196a-2 expression. Finally, ATXN1 and STC1 gene products have been identified as targets of miR-101b. Therefore, it is reasonable to hypothesize that HMGA1 proteins are involved in several functions by regulating miRNA expression.

Evaluation of magnetic resonance signal modification induced by hyaluronic acid therapy in chondromalacia patellae: a preliminary study.

Hyaluronic Acid (HA) is an alternative method for the treatment of osteoarthritis (OA), which acts on pain through a double action: anti-inflammatory and synovial fluid (SF) visco-supplementation. Magnetic Resonance Imaging (MRI), utilizing specific sequences, is a valid method for studying the initial phase of chondral damage. The analysis of the data, obtained through the intensity of values taken by positioning Region of Interest (ROIs) within the lesion, determining the differences before and after treatment with HA injected into the knee. The results obtained after six months and one year from the injection were statistically different in respect to those taken before, immediately and after three months of treatment. MRI represents a valid tool to evaluate the grade of chondromalacia patellae and also to follow the cartilage modification induced by HA therapy.

Analysis of UbcH10 expression represents a useful tool for the diagnosis and therapy of astrocytic tumors.

Previous studies suggest the expression of UbcH10 gene, that codes for a protein belonging to the ubiquitin-conjugating enzyme family, as a valid indicator of the proliferative and aggressive status of tumors of different origin. Therefore, to look for possible tools to be used as diagnostic markers in astrocytic neoplasias, we investigated UbcH10 expression in normal brain, gliosis and low-grade and high-grade astrocytic tumors by immunohistochemistry. UbcH10 expression was observed in low-grade astrocytoma and in glioblastoma. Our data indicate a clear correlation between UbcH10 expression and the histological grade of the astrocytic tumors. Moreover, the analysis of UbcH10 expression allows the differentiation between gliotic and malignant tissues. Finally, since proteasome inhibitors have recently been considered as possible drugs in the chemotherapy of various tumors, our results would suggest new perspectives for the treatment of brain malignancies based on the suppression of the UbcH10 function.

Procollagen I COOH-terminal fragment induces VEGF-A and CXCR4 expression in breast carcinoma cells.

The COOH-terminal fragment of procollagen type I (C3) is produced in tissues with high synthesis of collagen I, such as in breast cancer stroma and in bone. We previously demonstrated that C3 is chemoattractant for breast carcinoma and endothelial cells, and that in tumor cells it induces expression and activation of metalloproteinases (MMP) -2 and -9. Here we demonstrate that C3 induces expression of vascular-endothelial growth factor (VEGF) and of CXCR4, the receptor of the CXCL12/SDF-1 chemokine, in MDA MB 231 breast cancer cells. We show that the changes in gene expression and motility induced by C3 occur in a timely succession and are mediated by multiple and different signaling pathways. C3 induces early phosphorylation of p38/MAPK. Induction of VEGF expression requires continual activity of p38/MAPK and of Protein Kinase C (PKC). Pro-MMP-2 and -9 are induced through a signaling pathway involving G0alpha.i protein, and cell migration requires the activity of a combination of these signaling pathways. Our results suggest that C3 acts as a stromal-derived, cancer-promoting agent active in inducing the migratory phenotype and the survival of cancer cells and determining timely changes in their gene expression that establish conditions promoting tumor angiogenesis and invasion.

Isolation and characterization of microsatellite markers for Casearia sylvestris Sw. (Salicaceae), a neotropical medicinal tree.

Casearia sylvestris Sw. is a widespread neotropical tree utilized in popular medicine. Recent research ranked Casearia as one of the most promising genus in the search of drugs against cancer. Despite its wide distribution and pharmacological importance, no microsatellite markers have yet been developed for this genus. In this study, we provide 10 polymorphic microsatellite loci specifically designed for C. sylvestris, used to analyse 90 individuals distributed in two populations from São Paulo state, Brazil. On average, 12.3 alleles per locus were identified, showing the ability of the markers to detect microsatellite polymorphism in this species.