The human microbiomes in pancreatic cancer: Towards evidence-based manipulation strategies?

Recent pieces of evidence have emerged on the relevance of microorganisms in modulating responses to anticancer treatments and reshaping the tumor-immune microenvironment. On the one hand, many studies have addressed the role of the gut microbiota, providing interesting correlative findings with respect to etiopathogenesis and treatment responses. On the other hand, intra-tumoral bacteria are being recognized as intrinsic and essential components of the cancer microenvironment, able to promote a plethora of tumor-related aspects from cancer growth to resistance to chemotherapy. These elements will be probably more and more valuable in the coming years in early diagnosis and risk stratification. Furthermore, microbial-targeted intervention strategies may be used as adjuvants to current therapies to improve therapeutic responses and overall survival. This review focuses on new insights and therapeutic approaches that are dawning against pancreatic cancer: a neoplasm that arises in a central metabolic “hub” interfaced between the gut and the host

Characterization of Large Volume 3.5 x 8 inches LaBr3:Ce Detectors

The properties of large volume cylindrical 3.5 x 8 inches (89 mm x 203 mm) LaBr3:Ce scintillation detectors coupled to the Hamamatsu R10233-100SEL photo-multiplier tube were investigated. These crystals are among the largest ones ever produced and still need to be fully characterized to determine how these detectors can be utilized and in which applications. We tested the detectors using monochromatic gamma-ray sources and in-beam reactions producing gamma rays up to 22.6 MeV; we acquired PMT signal pulses and calculated detector energy resolution and response linearity as a function of gamma-ray energy. Two different voltage dividers were coupled to the Hamamatsu R10233-100SEL PMT: the Hamamatsu E1198-26, based on straightforward resistive network design, and the LABRVD, specifically designed for our large volume LaBr3:Ce scintillation detectors, which also includes active semiconductor devices. Because of the extremely high light yield of LaBr3:Ce crystals we observed that, depending on the choice of PMT, voltage divider and applied voltage, some significant deviation from the ideally proportional response of the detector and some pulse shape deformation appear. In addition, crystal non-homogeneities and PMT gain drifts affect the (measured) energy resolution especially in case of high-energy gamma rays. We also measured the time resolution of detectors with different sizes (from 1x1 inches up to 3.5x8 inches), correlating the results with both the intrinsic properties of PMTs and GEANT simulations of the scintillation light collection process. The detector absolute full energy efficiency was measured and simulated up to gamma-rays of 30 Me

Effets de la formation de la pourriture acide sur la composition de la fraction lipidique des différentes parties du grain de Vitis vinifera cv. Fortana

On a étudié par comparaison la composition de la fraction lipidique des différentes parties de grains de Vitis vinifera sains et attaqués par la pourriture acide.Des différences considérables ont été révélées entre les extraits des différents types d'échantillons de pulpe + peau en ce qui concerne les paramètres suivants: teneur en lipide, composition des triglycérides, composition des acides gras. Par contre, la fraction lipidique des deux échantillons de pépins était très semblable.Les composants principaux ainsi que leur répartition quantitative caractéristique correspondent aux résultats publiés dans une étude précédente. On a mis en évidence l'aldéhyde oléanoique qui n'était présente que dans quelques échantillons de pulpe + peau en même temps que l'acide correspondant et l'erythrodiol.Effects of sour rot on the composition of the lipid fraction of different parts of grapevine berries ( Vitis vinifera cv. Fortana)The composition of the lipid fraction of the various parts of berries of Vitis vinifera, healthy and infected by sour rot, have been compared. Notable differences became evident among the extracts of the various samples of pulp + skin with respect to the following parameters: lipid contens, composition of triglycerides and composition of fatty acids. The composition of the lipid fraction of the two samples of grape seeds, however, turned out to be very similar.The principal components and their quantitative distribution characteristics corresponded with the results reported in a preceding work. Oleanoic aldehyde has been identified which, together with the corresponding acid and erythrodiole, was only present in the samples of pulp + skin

Antitumoral efficacy of the protease inhibitor gabexate mesilate in colon cancer cells harbouring KRAS, BRAF and PIK3CA mutations

The employment of anti-epidermal growth factor receptor (EGFR) antibodies represents a backbone of the therapeutic options for the treatment of metastatic colorectal cancer (mCRC). However, this therapy is poorly effective or ineffective in unselected patients. Mutations in KRAS, BRAF and PIK3CA genes have recently emerged as the best predictive factors of low/absent response to EGFR-targeted therapy. Due to the need for efficacious treatment options for mCRC patients bearing these mutations, in this short report we examined the antitumoral activity of the protease inhibitor gabexate mesilate, alone and in combination with the anti-EGFR monoclonal antibody cetuximab, in a panel of human CRC cell lines harbouring a different expression pattern of wild-type/mutated KRAS, BRAF and PIK3CA genes. Results obtained showed that gabexate mesilate significantly inhibited the growth, invasive potential and tumour-induced angiogenesis in all the CRC cells employed in this study (including those ones harbouring dual KRAS/PIK3CA or BRAF/PIK3CA mutation), while cetuximab affected these parameters only in CRC cells with KRAS, BRAF and PIK3CA wild-type. Notably, the antitumoral efficacy of gabexate mesilate and cetuximab in combination was found to be not superior than that observed with gabexate mesilate as single agent. Overall, these preliminary findings suggest that gabexate mesilate could represent a promising therapeutic option for mCRC patients, particularly for those harbouring KRAS, BRAF and PIK3CA mutations, either as monotherapy or in addition to standard chemotherapy regimens. Further studies to better elucidate gabexate mesilate mechanism of action in CRC cells are therefore warranted

Rapid neuronal differentiation of induced pluripotent stem cells for measuring network activity on micro-electrode arrays

Neurons derived from human induced Pluripotent Stem Cells (hiPSCs) provide a promising new tool for studying neurological disorders. In the past decade, many protocols for differentiating hiPSCs into neurons have been developed. However, these protocols are often slow with high variability, low reproducibility, and low efficiency. In addition, the neurons obtained with these protocols are often immature and lack adequate functional activity both at the single-cell and network levels unless the neurons are cultured for several months. Partially due to these limitations, the functional properties of hiPSC-derived neuronal networks are still not well characterized. Here, we adapt a recently published protocol that describes production of human neurons from hiPSCs by forced expression of the transcription factor neurogenin-212. This protocol is rapid (yielding mature neurons within 3 weeks) and efficient, with nearly 100% conversion efficiency of transduced cells (>95% of DAPI-positive cells are MAP2 positive). Furthermore, the protocol yields a homogeneous population of excitatory neurons that would allow the investigation of cell-type specific contributions to neurological disorders. We modified the original protocol by generating stably transduced hiPSC cells, giving us explicit control over the total number of neurons. These cells are then used to generate hiPSC-derived neuronal networks on micro-electrode arrays. In this way, the spontaneous electrophysiological activity of hiPSC-derived neuronal networks can be measured and characterized, while retaining interexperimental consistency in terms of cell density. The presented protocol is broadly applicable, especially for mechanistic and pharmacological studies on human neuronal networks

INTRAHEPATIC CHOLANGIOCARCINOMA DEVELOPMENT IN A PATIENT WITH A NOVEL BAP1 GERMLINE MUTATION AND LOW EXPOSURE TO ASBESTOS

BRCA1 associated protein-1 (BAP1) germline mutations define a novel hereditary cancer syndrome, namely BAP1 tumor predisposition syndrome (BAP1-TPDS), characterized by an increased susceptibility to develop different cancer types, including mesothelioma, uveal and cutaneous melanoma, renal cell carcinoma, and basal cell and squamous cell carcinoma. Currently, the role of BAP1 germline mutations in intrahepatic cholangiocarcinoma (iCCA) pathogenesis is less known. Here we report the first clinical case of a female patient who developed an iCCA when she was 47-years-old and was found to carry a novel germline mutation at a splicing site of exon 4 in BAP1 gene (NM_004656.4: c.255_255+6del). An accurate anamnesis revealed the absence of risk factors linked to iCCA development, except for a low occupational exposure to asbestos. In tumor tissue, BAP1 sequencing, multiplex ligation-dependent probe amplification and immunoistochemistry showed the loss of heterozygosity and lack of nuclear expression, suggesting that BAP1 wild-type allele and functional protein were lost in cancer cells, in line with the classical two-hit model of tumor suppressor genes. Further studies are needed to confirm whether iCCA may be included into BAP1-TPDS cancer phenotypes and whether minimal asbestos exposure may facilitate the development of this malignancy in individuals carrying BAP1 germline mutations

Percutaneous radiofrequency ablation in intrahepatic cholangiocarcinoma: a retrospective single-center experience

Background & aims: Very few data are available in literature about the role of radiofrequency ablation (RFA) in intrahepatic cholangiocarcinoma (ICC) and previous studies are mainly case reports and case series on a very small number of patients and nodules. In this study, we aimed to evaluate effectiveness and safety of RFA for the treatment of unresectable ICC. Methods: This is a retrospective observational cohort study comprising all consecutive patients treated with RFA for unresectable ICC at Policlinico Sant’Orsola Malpighi Hospital, Bologna, Italy. Primary endpoint was Local Tumor Progression-Free Survival (LTPFS) while Overall Survival (OS) was also assessed as secondary endpoint. Results: From January 2014 to June 2019, 29 patients with 117 nodules underwent RFA. Technique effectiveness 1 month after RFA was 92.3%; median LTPFS was 9.27 months. Univariate analysis and multivariate analysis showed that LTPFS was significantly related to tumor size ≥20 mm. At a median follow up of 39.9 months, median OS from the date of RFA was 27.5 months, with an OS of 89%, 45% and 11% at 1, 2 and 4 years, respectively. Number of overall lesions and the sum of their diameter at the moment of the first RFA significantly affected OS in multivariate analysis. Minor and major complication rates were 14% and 7%, respectively. Conclusion: Tumor size ≥20 mm was associated with lower LTPFS, representing a potential useful threshold value. A careful evaluation of tumor burden appears as a crucial element in choosing the best therapeutic strategy in unresectable ICC

Characterization of Large Volume 3.5″ x 8″ LaBr3:Ce Detectors for the HECTOR+ array

A selection of the properties of large volume, cylindrical 3.5" x 8" LaBr 3 :Ce scintillation detectors coupled to a 3.5" PMT (model R10233-1000SEL from HAMAMATSU) and a special designed Voltage Divider (LABRVD) will be discussed. A number of 10 of such detectors constitute the HECTOR + array which, in fall 2012, measured at GSI coupled to the AGATA DEMOSTRATOR at the PRESPEC experimental setup. These crystals are among the largest ever produced and needed to be characterized. We have performed several tests and here we discuss, in particular, the energy resolution measured using monochromatic γ −ray sources and in-beam reactions producing γ −rays up to 22.6 MeV. As already measured in two previous works a saturation in the energy resolution was observed in case of high energy gamma rays. Crystal non-homogeneities and PMT gain drifts can affect the resolution of measurements especially in case of high energy γ −rays

Immunotherapy in Pancreatic Cancer: Why Do We Keep Failing? A Focus on Tumor Immune Microenvironment, Predictive Biomarkers and Treatment Outcomes

The advent of immunotherapy and targeted therapies has dramatically changed the outcomes of patients affected by many malignancies. Pancreatic cancer (PC) remains one the few tumors that is not treated with new generation therapies, as chemotherapy still represents the only effective therapeutic strategy in advanced-stage disease. Agents aiming to reactivate the host immune system against cancer cells, such as those targeting immune checkpoints, failed to demonstrate significant activity, despite the success of these treatments in other tumors. In many cases, the proportion of patients who derived benefits in early-phase trials was too small and unpredictable to justify larger studies. The population of PC patients with high microsatellite instability/mismatch repair deficiency is currently the only population that may benefit from immunotherapy; nevertheless, the prevalence of these alterations is too low to determine a real change in the treatment scenario of this tumor. The reasons for the unsuccess of immunotherapy may lie in the extremely peculiar tumor microenvironment, including distinctive immune composition and cross talk between different cells. These unique features may also explain why the biomarkers commonly used to predict immunotherapy efficacy in other tumors seem to be useless in PC. In the current paper, we provide a comprehensive and up-to-date review of immunotherapy in PC, from the analysis of the tumor immune microenvironment to immune biomarkers and treatment outcomes, with the aim to highlight that simply transferring the knowledge acquired on immunotherapy in other tumors might not be a successful strategy in patients affected by PC

Pregnancy outcome after loop electrosurgical excision procedure for cervical intraepithelial neoplasia

To determine pregnancy outcomes among women who underwent loop electrosurgical excision procedure (LEEP)