Organs on chip approach: A tool to evaluate cancer-immune cells interactions

In this paper we discuss the applicability of numerical descriptors and statistical physics concepts to characterize complex biological systems observed at microscopic level through organ on chip approach. To this end, we employ data collected on a micro uidic platform in which leukocytes can move through suitably built channels toward their target. Leukocyte behavior is recorded by standard time lapse imaging. In particular, we analyze three groups of human peripheral blood mononuclear cells (PBMC): heterozygous mutants (in which only one copy of the FPR1 gene is normal), homozygous mutants (in which both alleles encoding FPR1 are loss-of-function variants) and cells from ‘wild type’ donors (with normal expression of FPR1). We characterize the migration of these cells providing a quantitative con rmation of the essential role of FPR1 in cancer chemotherapy response. Indeed wild type PBMC perform biased random walks toward chemotherapy-treated cancer cells establishing persistent interactions with them. Conversely, heterozygous mutants present a weaker bias in their motion and homozygous mutants perform rather uncorrelated random walks, both failing to engage with their targets. We next focus on wild type cells and study the interactions of leukocytes with cancerous cells developing a novel heuristic procedure, inspired by Lyapunov stability in dynamical systems

Mammographic calcifications undergoing percutaneous biopsy: outcome in women with and without a personal history of breast cancer

PURPOSE To compare the positive predictive values (PPVs) of BI-RADS categories used to assess pure mammographic calcifications in women with and without a previous history of breast cancer (PHBC). MATERIALS AND METHODS In this retrospective study, all consecutive pure mammographic calcifications (n = 320) undergoing a stereotactic biopsy between 2016 and 2018 were identified. Mammograms were evaluated in consensus by two radiologists according to BI-RADS and blinded to patient history and pathology results. Final pathologic results were used as the standard of reference. PPV of BI-RADS categories were compared between the two groups. Data were evaluated using standard statistics, Mann-Whitney U tests and Chi-square tests. RESULTS Two hundred sixty-eight patients (274 lesions, median age 54 years, inter-quartile range, 50-65 years) with a PHBC (n = 46) and without a PHBC (n = 222) were included. Overall PPVs were the following: BI-RADS 2, 0% (0 of 56); BI-RADS 3, 9.1% (1 of 11); BI-RADS 4a, 16.2% (6 of 37); BI-RADS 4b, 37.5% (48 of 128); BI-RADS 4c, 47.3% (18 of 38) and BI-RADS 5, 100% (4 of 4). The PPV of BI-RADS categories was similar in patients with and without a PHBC (P = .715). Calcifications were more often malignant in patients with a PHBC older than 10 years (47.3%, 9 of 19) compared to 1-2 years (25%, 1 of 4), 2-5 years (20%, 2 of 10) and 5-10 years (0%, of 13) from the first breast cancer (P = .005). CONCLUSION PPV of mammographic calcifications is similar in women with or without PHBC when BI-RADS classification is strictly applied. A higher risk of malignancy was observed in patients with a PHBC longer than 10 years

Improving phenolic total content and monoterpene in mentha x piperita by using salicylic acid or methyl jasmonate combined with rhizobacteria inoculation

The effects of plant inoculation with plant growth-promoting rhizobacteria (PGPR) and those resulting from the exogenous application of salicylic acid (SA) or methyl jasmonte (MeJA) on total phenolic content (TPC) and monoterpenes in Mentha x piperita plants were investigated. Although the PGPR inoculation response has been studied for many plant species, the combination of PGPR and exogenous phytohormones has not been investigated in aromatic plant species. The exogenous application of SA produced an increase in TPC that, in general, was of a similar level when applied alone as when combined with PGPR. This increase in TPC was correlated with an increase in the activity of the enzyme phenylalanine ammonia lyase (PAL). Also, the application of MeJA at different concentrations in combination with inoculation with PGPR produced an increase in TPC, which was more relevant at 4 mM, with a synergism effect being observed. With respect to the main monoterpene concentrations present in peppermint essential oil (EO), it was observed that SA or MeJA application produced a significant increase similar to that of the combination with rhizobacteria. However, when plants were exposed to 2 mM MeJA and inoculated, an important increase was produced in the concentration on menthol, pulegone, linalool, limonene, and menthone concentrations. Rhizobacteria inoculation, the treatment with SA and MeJA, and the combination of both were found to affect the amount of the main monoterpenes present in the EO of M. piperita. For this reason, the expressions of genes related to the biosynthesis of monoterpene were evaluated, with this expression being positively affected by MeJA application and PGPR inoculation, but was not modified by SA application. Our results demonstrate that MeJA or SA application combined with inoculation with PGPR constitutes an advantageous management practice for improving the production of secondary metabolites from M. piperita.Fil: Cappellari, Lorena del Rosario. Universidad Nacional de Rio Cuarto. Facultad de Cs.exactas Fisicoquimicas y Naturales. Instituto de Biotecnologia Ambiental y Salud. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Cordoba. Instituto de Biotecnologia Ambiental y Salud.; ArgentinaFil: Santoro, Valeria Maricel. Instituto Max Planck Institut für Chemische Okologie; AlemaniaFil: Schmidt, Axel. Instituto Max Planck Institut für Chemische Okologie; AlemaniaFil: Gershenzon, Jonathan. Instituto Max Planck Institut für Chemische Okologie; AlemaniaFil: Banchio, Erika. Universidad Nacional de Rio Cuarto. Facultad de Cs.exactas Fisicoquimicas y Naturales. Instituto de Biotecnologia Ambiental y Salud. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Cordoba. Instituto de Biotecnologia Ambiental y Salud.; Argentin

Quantification of Lung Fibrosis in IPF-Like Mouse Model and Pharmacological Response to Treatment by Micro-Computed Tomography

Idiopathic pulmonary fibrosis (IPF) is a chronic progressive degenerative lung disease leading to respiratory failure and death. Although anti-fibrotic drugs are now available for treating IPF, their clinical efficacy is limited and lung transplantation remains the only modality to prolong survival of IPF patients. Despite its limitations, the bleomycin (BLM) animal model remains the best characterized experimental tool for studying disease pathogenesis and assessing efficacy of novel potential drugs. In the present study, the effects of oropharyngeal (OA) and intratracheal (IT) administration of BLM were compared in C57BL/6 mice. The development of lung fibrosis was followed in vivo for 28 days after BLM administration by micro-computed tomography and ex vivo by histological analyses (bronchoalveolar lavage, histology in the left lung to stage fibrosis severity and hydroxyproline determination in the right lung). In a separate study, the antifibrotic effect of Nintedanib was investigated after oral administration (60 mg/kg for two weeks) in the OA BLM model. Lung fibrosis severity and duration after BLM OA and IT administration was comparable. However, a more homogeneous distribution of fibrotic lesions among lung lobes was apparent after OA administration. Quantification of fibrosis by micro-CT based on % of poorly aerated tissue revealed that this readout correlated significantly with the standard histological methods in the OA model. These findings were further confirmed in a second study in the OA model, evaluating Nintedanib anti-fibrotic effects. Indeed, compared to the BLM group, Nintedanib inhibited significantly the increase in % of poorly aerated areas (26%) and reduced ex vivo histological lesions and hydroxyproline levels by 49 and 41%, respectively. This study indicated that micro-computed tomography is a valuable in vivo technology for lung fibrosis quantification, which will be very helpful in the future to better evaluate new anti-fibrotic drug candidates

Esophageal testing: What we have so far

Gastroesophageal reflux disease (GERD) is a common disorder of the gastrointestinal tract. In the last few decades, new technologies have evolved and have been applied to the functional study of the esophagus, allowing for the improvement of our knowledge of the pathophysiology of GERD. High-resolution manometry (HRM) permits greater understanding of the function of the esophagogastric junction and the risks associated with hiatal hernia. Moreover, HRM has been found to be more reproducible and sensitive than conventional water-perfused manometry to detect the presence of transient lower esophageal sphincter relaxation. Esophageal 24-h pH-metry with or without combined impedance is usually performed in patients with negative endoscopy and reflux symptoms who have a poor response to anti-reflux medical therapy to assess esophageal acid exposure and symptom-reflux correlations. In particular, esophageal 24-h impedance and pH monitoring can detect acid and non-acid reflux events. EndoFLIP is a recent technique poorly applied in clinical practice, although it provides a large amount of information about the esophagogastric junction. In the coming years, laryngopharyngeal symptoms could be evaluated with up and coming non-invasive or minimally invasive techniques, such as pepsin detection in saliva or pharyngeal pH-metry. Future studies are required of these techniques to evaluate their diagnostic accuracy and usefulness, although the available data are promising

Management of Asymptomatic Sporadic Nonfunctioning Pancreatic Neuroendocrine Neoplasms (ASPEN) <= 2 cm: Study Protocol for a Prospective Observational Study

Introduction: The optimal treatment for small, asymptomatic, nonfunctioning pancreatic neuroendocrine neoplasms (NF-PanNEN) is still controversial. European Neuroendocrine Tumor Society (ENETS) guidelines recommend a watchful strategy for asymptomatic NF-PanNEN <2 cm of diameter. Several retrospective series demonstrated that a non-operative management is safe and feasible, but no prospective studies are available. Aim of the ASPEN study is to evaluate the optimal management of asymptomatic NF-PanNEN ≤2 cm comparing active surveillance and surgery. Methods: ASPEN is a prospective international observational multicentric cohort study supported by ENETS. The study is registered in ClinicalTrials.gov with the identification code NCT03084770. Based on the incidence of NF-PanNEN the number of expected patients to be enrolled in the ASPEN study is 1,000 during the study period (2017–2022). Primary endpoint is disease/progression-free survival, defined as the time from study enrolment to the first evidence of progression (active surveillance group) or recurrence of disease (surgery group) or death from disease. Inclusion criteria are: age >18 years, the presence of asymptomatic sporadic NF-PanNEN ≤2 cm proven by a positive fine-needle aspiration (FNA) or by the presence of a measurable nodule on high-quality imaging techniques that is positive at 68Gallium DOTATOC-PET scan. Conclusion: The ASPEN study is designed to investigate if an active surveillance of asymptomatic NF-PanNEN ≤2 cm is safe as compared to surgical approach

Characterization of a new animal model of lysosomal storage disease: Gangliosidosis in a family of boars

Le patologie da accumulo lisosomiale (LSDs) sono un gruppo eterogeneo di rare, progressive, letali, multisistemiche patologie con un meccanismo ereditario autosomico recessivo. Le LSDs sono caratterizzate da una funzionalità deficitaria di enzimi lisosomiali specifici, causata da difetti genetici. A partire dalla via di degradazione lisosomiale strettamente sequenziale degli sfingolipidi, sono stati descritti difetti per quasi tutti i passaggi della loro degradazione; ciò determina un blocco metabolico e un accumulo di substrati corrispondenti non degradati. Queste patologie possono essere classificate secondo il difetto genetico, l’enzima deficitario o i prodotti accumulati. I gangliosidi sono uno dei maggiori componenti delle membrane neuronali e partecipano a processi cruciali per il sistema nervoso. Difetti genetici nel catabolismo dei gangliosidi ed il loro accumulo determinano una LSD neuronale, definita Gangliosidosi. Questa patologia può essere sottoclassificata in Gangliosidosi GM1, causata da una deficienza di β-galattosidasi con accumulo di ganglioside GM1, o Gangliosidosi GM2, causata da deficienza di β-esosaminidasi o proteina attivatrice GM2 con accumulo di ganglioside GM2. Poiché la patologia nell’uomo è relativamente rara, i modelli animali sono strumenti indispensabili per studi sulla patogenesi e per lo sviluppo di potenziali trattamenti. Lo scopo del progetto è di studiare le caratteristiche morfologiche e biochimiche di un caso di Gangliosidosi GM2 in tre cinghiali della stessa nidiata. I tre cinghiali, appartenenti ad un allevamento brado, hanno presentato sintomatologia neurologica (dismetria, atassia, quadriplegia e decubito laterale) a 6 mesi di età. Sono state eseguite analisi virologiche e batteriologiche, risultate negative per culture batteriche, Peste suina classica e Aujeszky; sono state inoltre escluse tossicosi indotte da piante e farmaci. A causa del peggioramento delle condizioni cliniche, i cinghiali sono stati sacrificati a circa un anno di età e sottoposti a indagini necroscopiche. Lesioni macroscopiche, comuni in tutti gli animali affetti, erano riduzione della consistenza del parenchima cerebrale e cerebellare, degenerazione epatica diffusa e dilatazione gastrointestinale. L’esame istologico ha rilevato nel cervello, cervelletto, midollo spinale, gangli periferici e retina, la presenza di neuroni aumentati di dimensioni, con citoplasma diffusamente e severamente vacuolizzato. Altre lesioni neuropatologiche includevano sferoidi, meganeuriti e microgliosi. I fenomeni di astrocitosi sono stati confermati con colorazioni immunoistochimiche per GFAP e vimentina; mentre la colorazione Luxol fast blu ha mostrato diffusi quadri di demielinizzazione. La microscopia elettronica ha evidenziato nel citoplasma dei neuroni la presenza di numerosi lisosomi, disposti singolarmente o in aggregati, ripieni di materiale membranoso organizzato in lamelle o vortici (corpi citoplasmatici membranosi). Studi biochimici hanno rivelato la presenza a livello cerebrale di un’elevata quantità di ganglioside GM2, confermando la diagnosi di Gangliosidosi GM2. Questa patologia fu descritta nel suino solamente nel 1978 nella razza Yorkshire (Sus scrofa domestica) e non è mai stata descritta in cinghiali selvatici (Sus scrofa). Questa forma di Gangliosidosi GM2, evidenziata per la prima volta in una nidiata di cinghiali, appare molto simile alla patologia riscontrata nell’uomo e può rappresentare un utile modello animale.Lysosomal storage diseases (LSDs) are heterogeneous group of rare, progressive, lethal, multisystemic diseases with autosomal recessive inheritance. LSDs are characterized by deficient function of specific lysosomal enzymes, due to a genetic defect. According to the strictly sequential lysosomal degradation pathway of glycosphingolipids, defects for almost every step in their degradation have been described; this lead to a block of degradation and accumulation of the corresponding undegraded substrates. These pathologies can be classified on the base of the underlying genetical defect, the affected enzyme or according to the accumulated products. Gangliosides are major components of neuronal membranes and participate in crucial processes of the nervous system. Genetic defects in catabolism of these glycosphingolipids and their accumulation cause a neuronal LSD, defined Gangliosidosis. These pathologies can be subclassified into GM1 gangliosidosis, caused by β-galactosidase deficiency with GM1 ganglioside accumulation, or GM2 gangliosidosis, caused by β-hexosaminidase or GM2 activator protein deficiency with GM2 ganglioside accumulation. Since the human diseases are relatively rare, animal models are indispensable tools for further study of pathogenesis and for development of potential treatments. The aim of the project is to study morphological and biochemical features of GM2 gangliosidosis in 3 wild boars of the same brood. Three littermate (3/3) wild boards, from a free ranging farm, presented neurological signs (dysmetria, ataxia, quadriplegia and lateral decubitus) at 6 months of age. Viral and bacterial analysis were performed and resulted negative for bacteria culture, Classical Swine Fever and Aujeszky; plants and drugs toxicosis were excluded. Due to the worsening conditions, they were euthanized at approximately one year of age and submitted for necropsy. Gross lesions, common in all affected animals, were reduction in consistency of cerebral and cerebellar parenchyma, diffuse hepatic degeneration and gastrointestinal dilation. Histology revealed in brain, cerebellum, spinal cord, peripheral ganglia and retina, enlarged foamy neurons, with diffusely severely vacuolated cytoplasm. Other neuropathological findings included spheroids, meganeurites and microgliosis. Astrocytosis was confirmed with Immunohistochemistry for GFAP and Vimentin; demyelination was evident with Luxol fast blue stain. Electron Microscopy analysis of neurons revealed that cytoplasm was enlarged by the presence of numerous lysosomes, singularly disposed or in aggregates, filled by membranous material arranged in lamellae and whorls (membranous cytoplasmic bodies). Biochemical studies revealed the presence of an elevate amount of GM2 ganglioside, confirming the diagnosis of GM2 gangliosidosis. GM2 gangliosidosis in swine has been described only in 1978 in purebred Yorkshire swine (Sus scrofa domestica) and has never been described in wild boars (Sus scrofa). This form of GM2 gangliosidosis in a family of wild boars is very similar to human disease and can be a potential useful animal model

GROSS EVALUATION, SAMPLING PROCEDURES AND HISTOLOGICAL CHARACTERIZATION OF FELINE MIDDLE EAR DISEASES

Introduction: Study of ear diseases is hampered by the difficult specimen preparation, resulting in the belief that these are uncommon in cats. The purpose of this study was to characterize gross and microscopic lesions of inflammatory and neoplastic origin, particularly of the middle ear. Materials and Methods: Ears from 26 cats were samplingduring necropsies, using a previously described technique (Sula et al. 2014) and were examined grossly and processed for routine histologic examination of the external, middle, and internal ear. Results: This sampling technique allows to have on a single slide the external, middle and internal ear andthis facilitates a single, complete and exhaustive evaluation of these structures. Gross and microscopic evaluation of feline ears highlighted numerous pathological conditions: 3 chronic external and medial otitis, 1 nasopharyngeal polyp, 1 ceruminous cystomatosis, 1 squamous cell carcinoma and 1 ceruminous adenocarcinoma with metastasis. Furthermore, we identified 1 case of mucoperiosteal exostoses. Conclusions: The sampling technique for the ear is simple to be performed and useful to evaluate all the anatomical structures. Focusing on middle ear alterations, these were frequently not reported in clinical history, but became evident during gross and, particularly, histopathological evaluations. We concluded that histologic evidence of middle ear diseases in cats is far greater than clinical literature reported; this suggests that ears, when possible, should be histologically evaluated. Furthermore, we reported for the first time mucoperiosteal exostoses in a domestic cat; this condition was described only in a single report in African lions