Lymph node fine needle cytology, Epstein Barr virus infection and Hodgkin Lymphoma

Epstein-Barr virus (EBV) is a double-strand DNA virus of the herpes family; it is one of the most common human viruses and it is associated with a wide spectrum of benign and malignant conditions. EBV is related to the development of several neoplasms, globally 1% of tumours, including lymphoproliferative, epithelial and mesenchymal neoplasm. Lymphoproliferative disorders include Hodgkin lymphoma (HL) and B and T cell non-Hodgkin lymphoma. HL is one of the most common lymphoma in the developed world, affecting both young people and adults. HL pathogenesis is complex and includes various and partially unknown mechanisms. EBV has been detected in some HL neoplastic cells and expresses genes with a potential oncogenic function, therefore many studies suggest that viral infections have a causative role in neoplastic transformation. Fine Needle Cytology (FNC) is extensively used in the first diagnosis of any lymph-nodal enlargement, including reactive lymphadeno - pathies and lymphoproliferative processes; therefore cytopathologists are likely to encounter EBVassociated malignancies in cytology samples, mainly HL, which is one of the most common lymphoma. This study focuses on the cytological features and ancillary studies required to diagnose EBV-related HL

Epidemiology and control of Ebola Virus Disease (EVD) in Sierra Leone: analysis of data from the Médecins Sans Frontières (MSF) response, 2014-15

Aims and Objectives This thesis stems from work conducted with Médecins Sans Frontières in Sierra Leone, during the 2014- 2015 Ebola virus disease (EVD) outbreaks. The thesis addresses 3 main public health knowledge gaps and aims to improve the response to future EVD outbreaks by: 1) Understanding the factors that influenced EVD transmission and community compliance with con- trol measures; 2) Estimating the performance of the WHO EVD case definition, and what components could improve it; 3) Estimating the design effect and mortality rates, and discussing how to improve future surveys for highly-clustered diseases. Methods The methods corresponding to each gap were: 1) A mixed-methods study in one village experiencing sustained transmission in Sierra Leone; 2) A review and meta-analysis exploring performance of WHO EVD case definitions against laboratory confirmed EVD; 3) Two population-based clustered surveys, in Sierra Leone. Results Study 1 identified that non-compliance with public health guidelines was a consequence of the failure of the response to orientate itself according to the needs and values of the community. Study 2 estimated that the WHO EVD case definition performed sub-optimally to identify cases (sen- sitivity 81·5% (74·1-87·2%); specificity 35·7% (28·5-43·6%)). Inclusion of intense fatigue as a key symptom and contact history could improve performance, but these changes would require collabora- tion with, and trust of, affected communities. Study 3 identified a high degree of clustering in community-based surveys of EVD, this contributed to imprecise mortality estimates, which have limited utility when assessing the impact of disease. Estimated the design effect along with methodological suggestions provided can inform future surveys for similar highly clustered diseases. Conclusion The thesis highlights that outbreak patterns are linked to social and cultural environments. Community influence key public health practices (i.e. case definitions). Recognition of transmission risks from responding organization requires a respectful and compassionate approach, understanding of social norms and adapted community-lead interventions

Neuroplastic Changes Following Brain Ischemia and their Contribution to Stroke Recovery: Novel Approaches in Neurorehabilitation

Ischemic damage to the brain triggers substantial reorganization of spared areas and pathways, which is associated with limited, spontaneous restoration of function. A better understanding of this plastic remodeling is crucial to develop more effective strategies for stroke rehabilitation. In this review article, we discuss advances in the comprehension of post-stroke network reorganization in patients and animal models. We first focus on rodent studies that have shed light on the mechanisms underlying neuronal remodeling in the perilesional area and contralesional hemisphere after motor cortex infarcts. Analysis of electrophysiological data has demonstrated brain-wide alterations in functional connectivity in both hemispheres, well beyond the infarcted area. We then illustrate the potential use of non-invasive brain stimulation (NIBS) techniques to boost recovery. We finally discuss rehabilitative protocols based on robotic devices as a tool to promote endogenous plasticity and functional restoration

Electrophysiology of glioma: a Rho GTPase-activating protein reduces tumor growth and spares neuron structure and function

Background. Glioblastomas are the most aggressive type of brain tumor. A successful treatment should aim at halting tumor growth and protecting neuronal cells to prevent functional deficits and cognitive deterioration. Here, we exploited a Rho GTPase-activating bacterial protein toxin, cytotoxic necrotizing factor 1 (CNF1), to interfere with glioma cell growth in vitro and vivo. We also investigated whether this toxin spares neuron structure and function in peritumoral areas. Methods. We performed a microarray transcriptomic and in-depth proteomic analysis to characterize the molecular changes triggered by CNF1 in glioma cells. We also examined tumor cell senescence and growth in vehicle-and CNF1-treated glioma-bearing mice. Electrophysiological and morphological techniques were used to investigate neuronal alterations in peritumoral cortical areas. Results. Administration of CNF1 triggered molecular and morphological hallmarks of senescence in mouse and human glioma cells in vitro. CNF1 treatment in vivo induced glioma cell senescence and potently reduced tumor volumes. In peritumoral areas of glioma-bearing mice, neurons showed a shrunken dendritic arbor and severe functional alterations such as increased spontaneous activity and reduced visual responsiveness. CNF1 treatment enhanced dendritic length and improved several physiological properties of pyramidal neurons, demonstrating functional preservation of the cortical network. Conclusions. Our findings demonstrate that CNF1 reduces glioma volume while at the same time maintaining the physiological and structural properties of peritumoral neurons. These data indicate a promising strategy for the development of more effective antiglioma therapies

Public health surveillance after the 2010 Haiti earthquake: the experience of Médecins Sans Frontières

Background In January 2010, Haiti was struck by a powerful earthquake, killing and wounding hundreds of thousands and leaving millions homeless. In order to better understand the severity of the crisis, and to provide early warning of epidemics or deteriorations in the health status of the population, Médecins Sans Frontières established surveillance for infections of epidemic potential and for death rates and malnutrition prevalence. Methods Trends in infections of epidemic potential were detected through passive surveillance at health facilities serving as sentinel sites. Active community surveillance of death rates and malnutrition prevalence was established through weekly home visits. Results There were 102,054 consultations at the 15 reporting sites during the 26 week period of operation. Acute respiratory infections, acute watery diarrhoea and malaria/fever of unknown origin accounted for the majority of proportional morbidity among the diseases under surveillance. Several alerts were triggered through the detection of immediately notifiable diseases and increasing trends in some conditions. Crude and under-5 death rates, and acute malnutrition prevalence, were below emergency thresholds. Conclusion Disease surveillance after disasters should include an alert and response component, requiring investment of resources in informal networks that improve sensitivity to alerts as well as on the more common systems of data collection, compilation and analysis. Information sharing between partners is necessary to strengthen early warning systems. Community-based surveillance of mortality and malnutrition is feasible but requires careful implementation and validation

Duplication of clostridial binding domains for enhanced macromolecular delivery into neurons

Neurological diseases constitute a quarter of global disease burden and are expected to rise worldwide with the ageing of human populations. There is an increasing need to develop new molecular systems which can deliver drugs specifically into neurons, non-dividing cells meant to last a human lifetime. Neuronal drug delivery must rely on agents which can recognise neurons with high specificity and affinity. Here we used a recently introduced ‘stapling’ system to prepare macromolecules carrying duplicated binding domains from the clostridial family of neurotoxins. We engineered individual parts of clostridial neurotoxins separately and combined them using a strong alpha-helical bundle. We show that combining two identical binding domains of tetanus and botulinum type D neurotoxins, in a sterically defined way by protein stapling, allows enhanced intracellular delivery of molecules into neurons. We also engineered a botulinum neurotoxin type C variant with a duplicated binding domain which increased enzymatic delivery compared to the native type C toxin. We conclude that duplication of the binding parts of tetanus or botulinum neurotoxins will allow production of high avidity agents which could deliver imaging reagents and large therapeutic enzymes into neurons with superior efficiency

Obese mice exposed to psychosocial stress display cardiac and hippocampal dysfunction associated with local brain-derived neurotrophic factor depletion

Introduction: Obesity and psychosocial stress (PS) co-exist in individuals of Western society. Nevertheless, how PS impacts cardiac and hippocampal phenotype in obese subjects is still unknown. Nor is it clear whether changes in local brain-derived neurotrophic factor (BDNF) account, at least in part, for myocardial and behavioral abnormalities in obese experiencing PS. Methods: In adult male WT mice, obesity was induced via a high-fat diet (HFD). The resident-intruder paradigm was superimposed to trigger PS. In vivo left ventricular (LV) performance was evaluated by echocardiography and pressure-volume loops. Behaviour was indagated by elevated plus maze (EPM) and Y-maze. LV myocardium was assayed for apoptosis, fibrosis, vessel density and oxidative stress. Hippocampus was analyzed for volume, neurogenesis, GABAergic markers and astrogliosis. Cardiac and hippocampal BDNF and TrkB levels were measured by ELISA and WB. We investigated the pathogenetic role played by BDNF signaling in additional cardiac-selective TrkB (cTrkB) KO mice. Findings: When combined, obesity and PS jeopardized LV performance, causing prominent apoptosis, fibrosis, oxidative stress and remodeling of the larger coronary branches, along with lower BDNF and TrkB levels. HFD/PS weakened LV function similarly in WT and cTrkB KO mice. The latter exhibited elevated LV ROS emission already at baseline. Obesity/PS augmented anxiety-like behaviour and impaired spatial memory. These changes were coupled to reduced hippocampal volume, neurogenesis, local BDNF and TrkB content and augmented astrogliosis. Interpretation: PS and obesity synergistically deteriorate myocardial structure and function by depleting cardiac BDNF/TrkB content, leading to augmented oxidative stress. This comorbidity triggers behavioral deficits and induces hippocampal remodeling, potentially via lower BDNF and TrkB levels. Fund: J.A. was in part supported by Rotary Foundation Global Study Scholarship. G.K. was supported by T32 National Institute of Health (NIH) training grant under award number 1T32AG058527. S.C. was funded by American Heart Association Career Development Award (19CDA34760185). G.A.R.C. was funded by NIH (K01HL133368-01). APB was funded by a Grant from the Friuli Venezia Giulia Region entitled: “Heart failure as the Alzheimer disease of the heart; therapeutic and diagnostic opportunities”. M.C. was supported by PRONAT project (CNR). N.P. was funded by NIH (R01 HL136918) and by the Magic-That-Matters fund (JHU). V.L. was in part supported by institutional funds from Scuola Superiore Sant'Anna (Pisa, Italy), by the TIM-Telecom Italia (WHITE Lab, Pisa, Italy), by a research grant from Pastificio Attilio Mastromauro Granoro s.r.l. (Corato, Italy) and in part by ETHERNA project (Prog. n. 161/16, Fondazione Pisa, Italy). Funding source had no such involvement in study design, in the collection, analysis, interpretation of data, in the writing of the report; and in the decision to submit the paper for publication

p27(Kip1 )is expressed in proliferating cells in its form phosphorylated on threonine 187

BACKGROUND: G1/S cell cycle progression requires p27(Kip1 )(p27) proteolysis, which is triggered by its phosphorylation on threonine (Thr) 187. Since its levels are abundant in quiescent and scarce in cycling cells, p27 is an approved marker for quiescent cells, extensively used in histopathology and cancer research. METHODS: However here we showed that by using a specific phosphorylation site (pThr187) antibody, p27 is detectable also in proliferative compartments of normal, dysplastic and neoplastic tissues. RESULTS: In fact, whereas un-phosphorylated p27 and MIB-1 showed a significant inverse correlation (Spearman R = -0.55; p < 0,001), pThr187-p27 was positively and significantly correlated with MIB-1 expression (Spearman R = 0.88; p < 0,001). Thus proliferating cells only stain for pThr187-p27, whereas they are un-reactive with the regular p27 antibodies. However increasing the sensitivity of the immunocytochemistry (ICH) by the use of an ultra sensitive detection system based on tiramide signal amplification, simultaneous expression and colocalisation of both forms of p27 was shown in proliferating compartments nuclei by double immunofluorescence and laser scanning confocal microscopy studies. CONCLUSION: Overall, our data suggest that p27 expression also occurs in proliferating cells compartments and the combined use of both regular and phospho- p27 antibodies is suggested