Transplantation of prodrug-converting neural progenitor cells for brain tumor therapy

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Bioassay-guided isolation of antiproliferative compounds from grape (Vitis vinifera) stems.

The fractionation, guided by cell-growth inhibition assay, of the EtOAc crude extract from grape stems of the Sicilian Vitis vinifera variety 'Nerello Mascalese' allowed identification of ten constituents, isolated either as pure compounds (1, 3–5, 7–10) or inseparable mixtures (2a-d and 6a-e). The pure constituents were: two triterpenoid acids, oleanolic (1) and betulinic acids (5); daucosterol (7); a stilbenoid, E-resveratrol (3) and its dimer E-ε-viniferin (4); the simple phenol gallic acid (8); and the flavanols catechin (9) and gallocatechin (10). Four 6′- O-acyldaucosterols (2a-d) and five 1,2-di- O-acyl-3- O-β-D-galactopyranosyl glycerols (6a-e) were also identified as inseparable mixtures. All the isolated compounds were subjected to spectroscopic analysis and MTT bioassay on MCF-7 human breast cancer cells. The majority showed growth-inhibitory activity, 5 being the most active (GI50 = 0.57 μM). Compounds 3–5 were also tested on HT-29, U-87-MG and U-373-MG cell lines

Anti-proliferative effects of Cetuximab and Trastuzumab in colorectal cancer cell lines

Colon cancer is one of the most common human malignancies and a leading cause of death worldwide. In Europe around 250.000 new colon cases are diagnosed each year, accounting for around 9% of all the malignancies (Labianca R et al., 2010; Berrino F et al., 2007). Dysregulation of the signalling pathways induced through EGF receptors (ErbB/ HER receptors) by their over-expression or constitutive activation can promote tumor processes (Lurje G et al., 2009), including colorectal cancer. Therefore, the ErbB/HER receptor family with their most prominent members EGFR and HER-2 represents validated targets for anti-cancer therapy. Cetuximab and trastuzumab are two monoclonal antibodies approved for treating, respectively, metastatic colorectal and breast cancer. Because the monotherapy with cetuximab in metastatic colorectal is often insufficient (Cunningham D et al., 2004), it is useful to develop complementary therapeutic strategies to enhance antibody efficacy. A possible approach is co-administration of inhibitors, targeting multiple members of the EGF receptor family. In this study we examined the effect of cetuximab and trastuzumab in combination using two human colon cancer cell lines as a model. We observed that the two drugs had a cytostatic effect and inhibited the proliferation of both the cell lines in a time- and concentration-dependent manner. However, the combination had lower efficacy on one cell line than the other, with growth inhibition of 31% in the former and 49% in the latter. This result was associated to specific changes in cell cycle distribution, while no apoptosis was observed. Chromosome copy number heterogeneity and aneuploidy in tumoral cell lines have been reported (Pellestor F et al., 1999). Our data deriving from the cell cycle analysis confirmed the aneuploidy and polyploidy in our cellular models and are useful to explain cellular response to the combination. We used fluorescent in situ hybridisation analysis to evaluate EGFR and HER-2 gene amplification status. Both the tumour cell lines resulted in an abnormal copy number for the two genes resulting from aneuploidy (polisomy of chromosome 7 and 17) which is not responsible for the difference in sensitivity to cetuximab and trastuzumab between the two cell lines. In order to understand and to improve the pharmacological efficacy of cetuximab and trastuzumab combination, it will be useful to elucidate the molecular mechanisms involved in their activity. This will allow to develop novel and interesting approaches to cancer therapy

Recent advances in molecular diagnostics of colorectal cancer by genomic arrays: proposal for a procedural shift in biological sampling and pathological report

Two forms of genetic instability have been described in colorectal cancer: chromosomal instability, characterized by structural and numerical chromosomal abnormalities and associated to aneuploidy; and microsatellite instability, characterized by a deficiency in the mismatch repair system that leads to slippage in microsatellites and is associated to euploidy. Thirteen colorectal cancer sample DNAs were analyzed after colectomy. High-resolution genome-wide DNA copy number and Single Nucleotide Polimorphism genotyping analysis was performed by Affymetrix SNP 6.0 arrays that interrogates 906,600 single nucleotide polymorphisms and 945,826 copy number probes. We implemented this analysis as part of a routine procedure that includes the sampling of fresh tissue from the tumor mass without affecting the subsequent standard histopathological procedure. The novel molecular technology allows the determination of a genome-wide molecular karyotype using only 500 ng of high-quality tumor DNA; it distinguishes the two main types of genomic instability, discriminating between chromosomal instability positive and negative tumors. It also detects loss of heterozygosity (LOH) regions, called copy neutral-LOH. Tumor-associated copy neutral-LOH regions may play a pivotal role in oncogenesis when they determine duplications of either activating or loss of function gene mutation. We observed recurrent gains of chromosomes 2, 7, 8q, 9, 12, 13, 20 and losses of chromosomes 4, 5, 8p, 15, 17p, 18, 22, and Y, in agreement with previous cytogenetic studies. The use of such sampling procedure could stimulate the routine detection of point mutations in specific genes, thus avoiding subsequent sectioning of formalin-fixed and paraffin-embedded samples

Immunohistochemistry evidence for gastric ghrelin cells hypertrophy in obese patients

Ghrelin cells are endocrine cells of the gastrointestinal tract mainly present in the glands of the stomach. They produce the “hunger hormone” that promotes appetite and adiposity in animal and human models. In response to these anabolic effects, several elements have suggested the influence of ghrelin on the regulation of metabolic functions and the development of obesity-related disorders. However, its physiologic significance is quite complex and some of the effects of ghrelin are still debated in the literature. Recent evidence suggests that ghrelin influences glucose homeostasis through the modulation of insulin secretion and insulin receptor signalling. T2 diabetes is the most frequent metabolic disorder associated with obesity. To date, the most effective treatment of obesity is bariatric surgery and one of the most used techniques is sleeve gastrectomy. It consists in the resection of 25% of the stomach producing a neo anatomy in which this organ becomes a tube-like structure, lacking the ghrelinrich part. Forty-seven percent of T2 diabetic obese patients treated by sleeve gastrectomy rescue their diabetic condition just after surgery and before any weight reduction. This clinical evidence clearly connects this new morphology of the stomach with the positive metabolic improvement and suggests that ghrelin could play a key role in this striking effect. We studied the gastric mucosa of the fundus of sleeve gastrectomized T2 diabetic-obese patients (n.5) by immunohistochemistry with Anti- Ghrelin antibody to verify the ghrelin cell morphology, immunoreactivity and density. Anti- Chromogranin A antibody was used to detect the whole endocrine population of gastric glands. We used the normal part of the gastric fundus removed from patients (n.3) suffering for gastric cancer, as control. Ghrelin receptor has been shown on peripheral nerve fibers of the autonomous nervous system, and because a direct innervation of Langerhans islets by intestinal neurons have been shown in the past, we extended our immunohistochemistry analyses to neuronal antigens in order to study the nervous network of gastric mucosae in these surgical samples. Results show that ghrelin cells in T2 diabetic obese patients represented a consistent percentage (48.4% vs 38% in controls) of chromogranin A immunoreactive cell population. Furthermore, ghrelin cells were hypertrophic and intensely immunostained when compared with those found in gastric mucosae of controls. Neuronal specific enolase immunoreactivity was clearly enhanced in the gastric mucosae of obese patients. These data suggest that the stomach of T2 diabetic obese patients increase ghrelin content and that sleeve gastrectomy could have an important metabolic role by anatomical reduction of this hormone. Furthermore, our result suggest a possible paracrine effect on the peripheral neuronal population that could also be implicated in this complex and striking clinical effect of bariatric surgery

PARP-14 Promotes Survival of Mammalian α but Not β Pancreatic Cells Following Cytokine Treatment

PARP-14 (poly-ADP Ribose Polymerase-14), a member of the PARP family, belongs to the group of Bal proteins (B Aggressive Lymphoma). PARP-14 has recently appeared to be involved in the transduction pathway mediated by JNKs (c Jun N terminal Kinases), among which JNK2 promotes cancer cell survival. Several pharmacological PARP inhibitors are currently used as antitumor agents, even though they have also proved to be effective in many inflammatory diseases. Cytokine release from immune system cells characterizes many autoimmune inflammatory disorders, including type I diabetes, in which the inflammatory state causes β cell loss. Nevertheless, growing evidence supports a concomitant implication of glucagon secreting α cells in type I diabetes progression. Here, we provide evidence on the activation of a survival pathway, mediated by PARP-14, in pancreatic α cells, following treatment of αTC1.6 glucagonoma and βTC1 insulinoma cell lines with a cytokine cocktail: interleukin 1 beta (IL-1β), interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α). Through qPCR, western blot and confocal analysis, we demonstrated higher expression levels of PARP-14 in αTC1.6 cells with respect to βTC1 cells under inflammatory stimuli. By cytofluorimetric and caspase-3 assays, we showed the higher resistance of α cells compared to β cells to apoptosis induced by cytokines. Furthermore, the ability of PJ-34 to modulate the expression of the proteins involved in the survival pathway suggests a protective role of PARP-14. These data shed light on a poorly characterized function of PARP-14 in αTC1.6 cells in inflammatory contexts, widening the potential pharmacological applications of PARP inhibitors

Computed Tomography Structured Reporting in the Staging of Lymphoma: A Delphi Consensus Proposal

Abstract Structured reporting (SR) in radiology is becoming increasingly necessary and has been recognized recently by major scientific societies. This study aims to build structured CT-based reports for lymphoma patients during the staging phase to improve communication between radiologists, members of multidisciplinary teams, and patients. A panel of expert radiologists, members of the Italian Society of Medical and Interventional Radiology (SIRM), was established. A modified Delphi process was used to develop the SR and to assess a level of agreement for all report sections. The Cronbach's alpha (Cα) correlation coefficient was used to assess internal consistency for each section and to measure quality analysis according to the average inter-item correlation. The final SR version was divided into four sections: (a) Patient Clinical Data, (b) Clinical Evaluation, (c) Imaging Protocol, and (d) Report, including n = 13 items in the "Patient Clinical Data" section, n = 8 items in the "Clinical Evaluation" section, n = 9 items in the "Imaging Protocol" section, and n = 32 items in the "Report" section. Overall, 62 items were included in the final version of the SR. A dedicated section of significant images was added as part of the report. In the first Delphi round, all sections received more than a good rating (≥3). The overall mean score of the experts and the sum of score for structured report were 4.4 (range 1-5) and 1524 (mean value of 101.6 and standard deviation of 11.8). The Cα correlation coefficient was 0.89 in the first round. In the second Delphi round, all sections received more than an excellent rating (≥4). The overall mean score of the experts and the sum of scores for structured report were 4.9 (range 3-5) and 1694 (mean value of 112.9 and standard deviation of 4.0). The Cα correlation coefficient was 0.87 in this round. The highest overall means value, highest sum of scores of the panelists, and smallest standard deviation values of the evaluations in this round reflect the increase of the internal consistency and agreement among experts in the second round compared to first round. The accurate statement of imaging data given to referring physicians is critical for patient care; the information contained affects both the decision-making process and the subsequent treatment. The radiology report is the most important source of clinical imaging information. It conveys critical information about the patient's health and the radiologist's interpretation of medical findings. It also communicates information to the referring physicians and records this information for future clinical and research use. The present SR was generated based on a multi-round consensus-building Delphi exercise and uses standardized terminology and structures, in order to adhere to diagnostic/therapeutic recommendations and facilitate enrolment in clinical trials, to reduce any ambiguity that may arise from non-conventional language, and to enable better communication between radiologists and clinicians

Fusion Transcripts of Adjacent Genes: New Insights into the World of Human Complex Transcripts in Cancer

The awareness of genome complexity brought a radical approach to the study of transcriptome, opening eyes to single RNAs generated from two or more adjacent genes according to the present consensus. This kind of transcript was thought to originate only from chromosomal rearrangements, but the discovery of readthrough transcription opens the doors to a new world of fusion RNAs. In the last years many possible intergenic cis-splicing mechanisms have been proposed, unveiling the origins of transcripts that contain some exons of both the upstream and downstream genes. In some cases, alternative mechanisms, such as trans-splicing and transcriptional slippage, have been proposed. Five databases, containing validated and predicted Fusion Transcripts of Adjacent Genes (FuTAGs), are available for the scientific community. A comparative analysis revealed that two of them contain the majority of the results. A complete analysis of the more widely characterized FuTAGs is provided in this review, including their expression pattern in normal tissues and in cancer. Gene structure, intergenic splicing patterns and exon junction sequences have been determined and here reported for well-characterized FuTAGs. The available functional data and the possible roles in cancer progression are discussed