Endometrial carcinomas with mismatch repair deficiency: immunophenotypical and molecular characterization.

This project was focused on the molecular and immunophenotypical characterization of a series of 80 gynecological cancers (GC, 64 endometrial, 12 ovarian, 4 endocervical) from patients who referred to the Cancer Genetic Counseling of Varese Hospital. We investigated the immunohistochemical (IHC) expression of MMR proteins, the microsatellite (MSI) profile of GC. We observed that the IHC analysis identified more cases with MMR deficit respect to the MSI analysis, in particular we identified a subgroup of cases with a borderline MSI profile. In order to improve MSI test, we tested four additional MSI target loci (RPL22, SRPR, MBD6 and NRIP) by setting up the MSI test in our series, these analysis revealed RPL22 and SRPR as informative loci in borderline cases and provided MSI evaluation criteria. We also analyzed MLH1 promoter methylation profile in MLH1 IHC negative samples to distinguish the sporadic MLH1 negative samples to LS cases. We set up the IHC analysis of ARID1A protein. We observed ARID1A protein loss in LS cancers, respect to sporadic cancer. Finally, we investigated the mutational landscape of 16 genes involved in endometrial cancer by targeted exome sequencing in 35 endometrial cancers. The genes most frequently mutated were ARID1A, ARID2, PTEN. The MSI cases show a higher mutational rate respect to MSS samples

Skeletal Myogenic Progenitors Originating from Embryonic Dorsal Aorta Coexpress Endothelial and Myogenic Markers and Contribute to Postnatal Muscle Growth and Regeneration

Skeletal muscle in vertebrates is derived from somites, epithelial structures of the paraxial mesoderm, yet many unrelated reports describe the occasional appearance of myogenic cells from tissues of nonsomite origin, suggesting either transdifferentiation or the persistence of a multipotent progenitor. Here, we show that clonable skeletal myogenic cells are present in the embryonic dorsal aorta of mouse embryos. This finding is based on a detailed clonal analysis of different tissue anlagen at various developmental stages. In vitro, these myogenic cells show the same morphology as satellite cells derived from adult skeletal muscle, and express a number of myogenic and endothelial markers. Surprisingly, the latter are also expressed by adult satellite cells. Furthermore, it is possible to clone myogenic cells from limbs of mutant c-Met-/- embryos, which lack appendicular muscles, but have a normal vascular system. Upon transplantation, aorta-derived myogenic cells participate in postnatal muscle growth and regeneration, and fuse with resident satellite cells. The potential of the vascular system to generate skeletal muscle cells may explain observations of nonsomite skeletal myogenesis and raises the possibility that a subset of satellite cells may derive from the vascular system

Video based valve motion combined with Computational Fluid Dynamics gives stable and accurate simulations of blood flow in the Realheart® Total Artificial Heart

Background: Patients with end-stage, biventricular heart failure, and for whom heart transplantation is not an option, may be given a Total Artificial Heart (TAH). The Realheart® is a novel TAH which pumps blood by mimicking the native heart with translation of an atrioventricular plane. The aim of this work was to create a strategy for using Computational Fluid Dynamics (CFD) to simulate haemodynamics in the Realheart®, including motion of the atrioventricular plane and valves. Methods: The accuracies of four different computational methods for simulating fluid-structure interaction of the prosthetic valves were assessed by comparison of chamber pressures and flow rates with experimental measurements. The four strategies were: prescribed motion of valves opening and closing at the atrioventricular plane extrema; simulation of fluid-structure interaction of both valves; prescribed motion of the mitral valve with simulation of fluid-structure interaction of the aortic valve; motion of both valves prescribed from video analysis of experiments. Results: The most accurate strategy (error in ventricular pressure of 6%, error in flow rate of 5%) used video-prescribed motion. With the Realheart operating at 80 bpm, the power consumption was 1.03 W, maximum shear stress was 15 Pa, and washout of the ventricle chamber after 4 cycles was 87%.Conclusions: This study, the first CFD analysis of this novel TAH, demonstrates that good agreement between computational and experimental data can be achieved. This method will therefore enable future optimisation of the geometry and motion of the Realheart®

Prospective comparative study of ERCP brush cytology and EUS-FNA for the diferential diagnosis of biliary strictures

OBJECTIVE: To evaluate and to compare the diagnostic yield of ERCP brush cytology (ERCP) and EUS-FNA in patients with biliary strictures and evaluates the agreement between general pathologists (GP) and expert GI pathologists (GIP) in the final diagnosis of biliary strictures. METHODS: Patients with biliary strictures documented by ERCP were included. Brush cytology was performed and during EUS, only visible mass lesions or localized bile duct wall thickening were aspirated. The gold standard method for diagnosis was surgical histology and/or follow-up. Tissue sampling results were: malignant, suspicious, atypical, insufficiently or benign. Specimens were interpreted by GP and GIP, blinded for prior tests results. RESULTS: 46 patients were included. Final diagnosis was malignancy in 37 (26 pancreatic - 11 biliary) and benign in 9 (8 chronic pancreatitis - 1 common bile duct inflammatory stricture). Sensitivity and accuracy for ERCP brush cytology were 43.2% and 52.2% for GP and 51.4% and 58.7% for GIP. Sensitivity and accuracy for EUS-FNA were 52.8% and 58.5%, respectively for GP and 69.4% e 73.2% for GIP. In comparison, the combination of brush cytology and EUS-FNA demonstrated higher sensitivity and accuracy for both GP (64.9% and 69.6%, respectively) and GIP (83.8% and 84.8%, respectively) and improved agreement with final diagnosis for both (mostly for GIP). CONCLUSION: Both, ERCP brush cytology and EUS-FNA has a similar yield for the diagnosis of biliary strictures. However, the combination of these methods results in an improved diagnostic accuracy. In addition, GIP might be expected to interpret specimens with greater accuracy than GP.OBJETIVO: Avaliar o desempenho diagnóstico da citologia obtida pela CPER, aquele obtido pela EE-PAAF e a concordância entre patologistas gerais (PG) e especialistas (PE) em pacientes com estenose biliar. MÉTODOS: Incluímos pacientes com estenose biliar identificados pela CPER. A EE-PAAF foi realizada apenas em áreas com efeito de massa ou da parede espessada do ducto biliar. O padrão-ouro foi a cirurgia, histologia e/ou o seguimento. As amostras teciduais foram consideradas: malignas, suspeitas, atípicas, insuficientes ou benignas. Os espécimes obtidos por cada método foi interpretado (cego) por um PG e outro PE. RESULTADO: 46 pacientes foram incluídos (37 malignos e 9 benignos). O diagnóstico final foi de tumor pancreático (26), biliar (11), pancreatite crônica (8) e estenose inflamatória do ducto biliar (1). Sensibilidade e acurácia da CPER foram 43,2% e 52,2% para o PG e 51,4% e 58,7% para o PE. Sensibilidade e acurácia da EE-PAAF foi 52,8% e 58,5% para o PG e 69,4% e 73,2% para o PE. A combinação entre a CPER e EE-PAAF demonstrou maior sensibilidade e acurácia para ambos PG (64,9% e 69,6%) e PE (83,8% e 84,8%), respectivamente. CONCLUSÃO: A citologia obtida pelo escovado da via biliar durante a CPER e as amostras teciduais colhidas pela EE-PAAF tem rendimento semelhante para o diagnóstico das estenoses biliares. No entanto, a combinação dos métodos resulta em uma maior acurácia. Além disso, espera-se que a interpretação das amostras ocorra com maior precisão pelo PE se comparado ao PG.Universidade de São Paulo Faculdade de Medicina de Ribeirão Preto Departamento de Cirurgia e AnatomiaUniversidade Federal de São Paulo (UNIFESP) Unidade de Diagnóstico AnatomopatológicoUNIFESP, Unidade de Diagnóstico AnatomopatológicoSciEL

Tumor escape and progression of HER-2/neu negative breast cancer under immune pressure

<p>Abstract</p> <p>Background</p> <p>Emerging data from pre-clinical and clinical studies suggest that HER-2/neu-specific T cell responses could induce HER-2/neu antigen loss in the tumor cells. These data suggest that patients with HER-2/neu negative breast cancer might have had HER-2/neu positive premalignant lesions in the past that progressed to HER-2/neu negative breast cancer under HER-2/neu-specific immune pressure.</p> <p>Methods</p> <p>We conducted a pilot study in patients with HER-2/neu positive and HER-2/neu negative breast cancers as well as a patient with ductal carcinoma in situ (DCIS). HER-2/neu expression was determined by FISH. HER-2/neu-specific T cell responses were determined by using IFN-γ ELISA. Expression of IFN-γ Rα in the tumors was determined by immunohistochemistry analysis of paraffin-embedded tissues.</p> <p>Results</p> <p>We determined that majority of (10 of 12) patients with HER-2/neu negative breast cancer had HER-2/neu-specific IFN-γ producing T cell responses which was stronger than those in patients with HER-2/neu positive tumors. Such immune responses were associated with nuclear translocation of IFN-γ Rα in their tumor cells. Patient with DCIS also showed HER-2/neu-specific T cell responses.</p> <p>Conclusion</p> <p>These data suggest that conducting retrospective studies in patients with HER-2/neu negative breast cancers and prospective studies in patients with HER-2/neu positive DCIS can determine whether HER-2/neu negative invasive carcinomas arise from HER-2/neu positive DCIS under the immune pressure.</p

Lymphocyte proliferation and apoptosis of lymphocyte subpopulations in bovine leukemia virus-infected dairy cows with high and low proviral load

Bovine leukemia virus (BLV) is one of the most important virus in dairy cattle. The infection behavior follows what we call the iceberg phenomenon: 60% of infected animals do not show clinical signs; 30% develop persistent lymphocytosis (PL); and the remaining 10%, die due to lymphosarcoma. BLV transmission depends on infected cell exchange and thus, proviral load is determinant. Understanding the mechanisms by which cattle governs the control of viral dissemination will be desirable for designing effective therapeutic or preventive strategies for BLV. The development of high proviral load (HPL) or low proviral load (LPL) might be associated to genetic factors and humoral immune responses, however cellular responses are not fully described. It is known that BLV affects cellular homeostasis: proliferation and apoptosis. It is also known that the BLV tropism is directed towards B lymphocytes, and that lymphocytotic animals have elevated amounts of these cells. Usually, when an animal is infected by BLV, the B markers that increase are CD21, CD5 and CD11b. This increase could be related to the modulation of apoptosis in these cells. This is the first work in which animals infected with BLV are classified according to their proviral load and the subpopulations of B and T lymphocytes are evaluated in terms of their percentage in peripheral blood and its stage of apoptosis and viability. PBMCs from HPL animals proliferated more than LPL and non-infected animals. CD11b+/CD5+ lymphocytes in LPL animals presented greater early and late apoptosis than HPL animals and cells of HPL animals had increased viability than LPL animals. Our results confirm that BLV alters the mechanism of apoptosis and proliferation of infected cells.Fil: Nieto Farías, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Souza, Fernando Nogueira. Universidade de Sao Paulo; BrasilFil: Lendez, Pamela Anahí. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Martinez Cuesta, Lucia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Santos, Kamila Reis. Universidade de Sao Paulo; BrasilFil: Della Libera, Alice Maria Melville Paiva. Universidade de Sao Paulo; BrasilFil: Ceriani, Maria Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Dolcini, Guillermina Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentin

332 Clinical and prognostic significance of junctional late gadolinium enhancement in patients with non-ischaemic cardiomyopathy

Abstract Aims Pulmonary hypertension (PH) carries a poor prognosis in patients with non-ischaemic dilated cardiomyopathy (NIDC). Cardiac magnetic resonance (CMR) with late gadolinium enhancement (LGE) evaluation can identify myocardial abnormalities. In particular, junctional LGE is already an established marker of adverse right ventricular (RV) remodelling in patients with pre-capillary PH. This study sought to assess the prevalence of junctional LGE by CMR in NIDC, its relationship with hemodynamic parameters and, moreover, its prognostic significance. Methods and results Patients with NIDC who underwent right heart catheterization (RHC) and CMR within 3 months in a tertiary hospital were enrolled. Patients with acute heart failure were excluded. Among others, RV and left ventricular (LV) volumes, junctional LGE at CMR, pulmonary artery pressure (PAP) and pulmonary capillary wedge pressure (PCWP) at RHC were tabulated. Pulmonary hypertension was defined accordingly to current Guidelines (median PAP at RHC ≥ 25 mmHg). The primary endpoint consisted of heart failure (HF) hospitalization during follow-up. A total of 188 patients [median age 49 (SD 15), 71% males] were evaluated. At morpho-functional CMR evaluation, most subjects (76%) had important systolic dysfunction (LV EF ≤ 35%). Junctional LGE was observed in 83 (44%) patients. Among patients with junctional LGE, 21 had LGE confined only to the junctional region, while 61 had also mid-wall interventricular septal stria and 21 a mid-wall stria in the lateral free LV wall. Patients with junctional LGE had lower RV EF (49% vs. 56%, P &lt; 0.001) and LV EF (27% vs. 30%, P = 0.012) when compared to those without junctional LGE although no differences in LV and RV dimensions were found. RHC showed PH in 83 patients (44%). Patients with junctional LGE showed a worse hemodynamic profile in terms of PH (55% vs. 36%; P = 0.011) and increase in PCWP (PCWP &gt; 15 mmHg in 60% vs. 42%; P = 0.015) compared to subjects without junctional LGE. Among 79 patients with PH and PCWP &gt; 15 mmHg, 75 (95%) had a combined post capillary and pre-capillary PH (diastolic pressure gradient ≥7 mmHg). Univariate analysis showed that junctional LGE was associated with a worse hemodynamic profile; on multivariable model, RV EF was significantly associated with the presence of junctional LGE (OR: 0.91; 95% CI: 0.87–0.96, P &lt; 0.001). During a median follow-up of 58 months, 33 patients (18%) died or underwent heart transplantation/ventricular assist device implantation, 17% in the junctional LGE group vs. 18% among those without junctional LGE. Thirty-eight patients (20%) had at least one episode of HF, 22 among junctional LGE group and 16 in control group (27% vs. 15%, P = 0.056). When adjusted for age, junctional LGE resulted a significant determinant of HF hospitalization (OR: 2.13, 95% CI: 1.02–4.44, P = 0.044). Conclusions Junctional LGE is detectable in almost half of NIDC patients and it is related to a worse haemodynamic profile, characterized by PH and elevated PCWP. Moreover, after adjustment for age, it was a significant determinant of HF hospitalization during follow-up in our population. Junctional LGE can therefore represent a useful prognostic tool, as marker of adverse ventricular remodelling likely related to ventricular interdependence

Pneumolysin binds to the mannose receptor C type 1 (MRC-1) leading to anti-inflammatory responses and enhanced pneumococcal survival

Streptococcus pneumoniae (the pneumococcus) is a major cause of mortality and morbidity globally, and the leading cause of death in children under 5 years old. The pneumococcal cytolysin pneumolysin (PLY) is a major virulence determinant known to induce pore-dependent pro-inflammatory responses. These inflammatory responses are driven by PLY–host cell membrane cholesterol interactions, but binding to a host cell receptor has not been previously demonstrated. Here, we discovered a receptor for PLY, whereby pro-inflammatory cytokine responses and Toll-like receptor signalling are inhibited following PLY binding to the mannose receptor C type 1 (MRC-1) in human dendritic cells and mouse alveolar macrophages. The cytokine suppressor SOCS1 is also upregulated. Moreover, PLY–MRC-1 interactions mediate pneumococcal internalization into non-lysosomal compartments and polarize naive T cells into an interferon-γlow, interleukin-4high and FoxP3+ immunoregulatory phenotype. In mice, PLY-expressing pneumococci colocalize with MRC-1 in alveolar macrophages, induce lower pro-inflammatory cytokine responses and reduce neutrophil infiltration compared with a PLY mutant. In vivo, reduced bacterial loads occur in the airways of MRC-1-deficient mice and in mice in which MRC-1 is inhibited using blocking antibodies. In conclusion, we show that pneumococci use PLY–MRC-1 interactions to downregulate inflammation and enhance bacterial survival in the airways. These findings have important implications for future vaccine design

Agenesis of the seventh cervical vertebra with spinal cord compression in a goat

A 4-year-old male goat was presented to the Hospital of Bovines and Small Ruminants, University of São Paulo - Brazil, showing fasciculation in all limbs, ataxia progressing to paralysis and opisthotonos. After determination of the vital parameters and the specific evaluation of the nervous system, sensitivity in the region corresponding to the sixth cervical vertebrae (C6) and first thoracic vertebrae (T1) was observed on palpation. Further investigation using X-ray, myelography, and ultrasound revealed the decrease of the intervertebral space between C6 and T1, the presence of spondylosis and the absence of the seventh cervical vertebra (C7), which contained only the vertebral arch and local spinous process and the compression of the spinal cord. Goat congenital malformations are underdiagnosed, therefore, requires further discussion, and studies regarding the genetic variations