Vascular wall stem cells. Selection and conditioning of progenitors useful for cell therapy. A pathological case study

The arterial wall contains MSCs with mesengenic and angiogenic abilities. These multipotent precursors have been isolated from variously-sized human adult segments, belying the notion that vessel wall is a relatively quiescent tissue. Recently, our group identified in normal human arteries a vasculogenic niche and subsequently isolated and characterized resident MSCs (VW-MSCs) with angiogenic ability and multilineage potential. To prove that VW-MSCs are involved in normal and pathological vascular remodeling, we used a long-term organ culture system; this method was of critical importance to follow spontaneous 3-D vascular remodeling without any influence of blood cells. Next we tried to identify and localize in situ the VW-MSCs and to understand their role in the vascular remodeling in failed arterial homografts. Subsequently, we isolated this cell population and tested in vitro their multilineage differentiation potential through immunohistochemical, immunofluorescence, RT-PCR and ultrastructural analysis. From 25-30cm2 of each vascular wall homograft sample, we isolated a cell population with MSCs properties; these cells expressed MSC lineage molecules (CD90, CD44, CD105, CD29, CD73), stemness (Notch-1, Oct-4, Sca-1, Stro-1) and pericyte markers (NG2) whilst were negative for hematopoietic and endothelial markers (CD34, CD133, CD45, KDR, CD146, CD31 and vWF). MSCs derived from failed homografts (H-MSCs) exhibited adipogenic, osteogenic and chondrogenic potential but scarce propensity to angiogenic and leiomyogenic differentiation. The present study demonstrates that failed homografts contain MSCs with morphological, phenotypic and functional MSCs properties; H-MSCs are long-lived in culture, highly proliferating and endowed with prompt ability to differentiate into adipocytes, osteocytes and chondrocytes; compared with VW-MSCs from normal arteries, H-MSCs show a failure in angiogenic and leiomyogenic differentiation. A switch in MSCs plasticity could be the basis of pathological remodeling and contribute to aneurysmal failure of arterial homografts. The study of VW-MSCs in a pathological setting indicate that additional mechanisms are involved in vascular diseases; their knowledge will be useful for opening new therapeutic options in cardiovascular diseases.

Computational Modeling of Cardiac Excitation-Contraction Coupling in Physiological and Pathological Conditions

The cardiomyocyte is a complex biological system where many mechanisms interact non-linearly to regulate the coupling between electrical excitation and mechanical contraction. For this reason, the development of mathematical models is fundamental in the field of cardiac electrophysiology, where the use of computational tools has become complementary to the classical experimentation. My doctoral research has been focusing on the development of such models for investigating the regulation of ventricular excitation-contraction coupling at the single cell level. In particular, the following researches are presented in this thesis: 1) Study of the unexpected deleterious effect of a Na channel blocker on a long QT syndrome type 3 patient. Experimental results were used to tune a Na current model that recapitulates the effect of the mutation and the treatment, in order to investigate how these influence the human action potential. Our research suggested that the analysis of the clinical phenotype is not sufficient for recommending drugs to patients carrying mutations with undefined electrophysiological properties. 2) Development of a model of L-type Ca channel inactivation in rabbit myocytes to faithfully reproduce the relative roles of voltage- and Ca-dependent inactivation. The model was applied to the analysis of Ca current inactivation kinetics during normal and abnormal repolarization, and predicts arrhythmogenic activity when inhibiting Ca-dependent inactivation, which is the predominant mechanism in physiological conditions. 3) Analysis of the arrhythmogenic consequences of the crosstalk between β-adrenergic and Ca-calmodulin dependent protein kinase signaling pathways. The descriptions of the two regulatory mechanisms, both enhanced in heart failure, were integrated into a novel murine action potential model to investigate how they concur to the development of cardiac arrhythmias. These studies show how mathematical modeling is suitable to provide new insights into the mechanisms underlying cardiac excitation-contraction coupling and arrhythmogenesis.

Statistical shape modeling of pathological scoliotic vertebrae: A comparative analysis

Statistical shape models (SSMs) have been used widely as a basis for segmenting and interpreting complex anatomical structures. The robustness of these models are sensitive to the registration procedures, i.e., establishment of a dense correspondence across a training data set. In this work, two SSMs based on the same training data set of scoliotic vertebrae, and registration procedures were compared. The first model was constructed based on the original binary masks without applying any image pre- and post-processing, and the second was obtained by means of a feature preserving smoothing method applied to the original training data set, followed by a standard rasterization algorithm. The accuracies of the correspondences were assessed quantitatively by means of the maximum of the mean minimum distance (MMMD) and Hausdorf distance (H(D)). Anatomical validity of the models were quantified by means of three different criteria, i.e., compactness, specificity, and model generalization ability. The objective of this study was to compare quasi-identical models based on standard metrics. Preliminary results suggest that the MMMD distance and eigenvalues are not sensitive metrics for evaluating the performance and robustness of SSMs.

Clinical, laboratory and pathological findings in dogs experimentally infected with Angiostrongylus vasorum

The aim of this comparative study was to investigate the development of clinical signs and accompanying haematological, coproscopic and pathological findings as a basis for the monitoring of health condition of Angiostrongylus vasorum infected dogs. Six beagles were orally inoculated with 50 (n=3) or 500 (n=3) A. vasorum third stage larvae (L3) obtained from experimentally infected Biomphalaria glabrata snails. Two dogs were treated with moxidectin/imidacloprid spot-on solution and two further dogs with an oral experimental compound 92 days post infection (dpi), and were necropsied 166 dpi. Two untreated control dogs were necropsied 97 dpi. Prepatency was 47-49 days. Dogs inoculated with 500 L3 exhibited earlier (from 42 dpi) and more severe respiratory signs. Clinical signs resolved 12 days after treatment and larval excretion stopped within 20 days in all four treated dogs. Upon necropsy, 10 and 170 adult worms were recovered from the untreated dogs inoculated with 50 and 500 L3, respectively. Adult worms were also found in two treated dogs, in the absence of L1 or eggs. Despite heavy A. vasorum infection load and severe pulmonary changes including vascular thrombosis, only mild haematological changes were observed. Eosinophilia was absent but the presence of plasma cells was observed. Neutrophilic leucocytes showed a transient increase but only after treatment. Signs for coagulopathies were slight; nevertheless coagulation parameters were inoculation dose dependent. Ten weeks after treatment pulmonary fibrosis was still present. Infections starting from 50 L3 of A. vasorum had a massive impact on lung tissues and therefore on the health of affected dogs, particularly after prepatency, although only mild haematological abnormalities were evident.

Virological and pathological findings in Bluetongue virus serotype 8 infected sheep

Twenty-seven sheep of the four most common Swiss breeds and the English breed Poll Dorset were experimentally infected with a northern European field strain of bluetongue virus serotype 8 (BTV-8). Animals of all breeds developed clinical signs, viremia and pathological lesions, demonstrating that BTV-8 is fully capable of replicating and inducing bluetongue disease (BT) in the investigated sheep. Necropsy performed between 10 and 16 days post-infectionem (d.p.i.) revealed BT-typical hemorrhages, effusions, edema, erosions and activation of lymphatic tissues. Hemorrhages on the base of the Arteria pulmonalis and the left Musculus papillaris subauricularis were frequently present. Histology confirmed the macroscopical findings. Using a score system, clinical manifestation and pathology were found to be significantly related. Furthermore, clinical signs and fever were shown to be indicative for the concurrent presence of high amounts of viral ribonucleic acid (RNA) in blood. Spleen, lung, lymph nodes and tonsils from all animals were analyzed regarding viral RNA loads and infectivity using real-time reverse transcriptase PCR (rRT-PCR) and virus isolation in cell culture, respectively. The highest amount of viral RNA was detected in spleen and lung and rRT-PCR revealed to be a more sensitive method for virus detection compared to virus isolation. A long-term follow-up was performed with three sheep showing that BTV-8 viral RNA in blood was present up to 133 d.p.i. and in certain tissues even on 151 d.p.i. No significant breed-related differences were observed concerning clinicopathological picture and viremia, and the Swiss sheep were as susceptible to BTV-8 infection as Poll Dorset sheep, demonstrating a remarkably high virulence of BTV-8 for indigenous sheep breeds.

Public Drinking in the Early Modern World: Voices from the Tavern, 1500-1800

Until recently the role of the public drinking house has been approached from elitist, folkloric and anecdotal perspectives. The work of a new generation of social historians, however, has raised the tavern’s profile in the academic consciousness and confirmed its position within the mainstream of social and cultural history. It is now recognized that an understanding of the centrality of public drinking to the development of both elite and popular culture is vital to studies of social behaviour. The study of taverns has also been at the forefront of emerging interest in the history of consumption and material culture, and has contributed to a richer understanding of economic history. Constructions of gender and identity are also visible through research into the patterns of behaviour and discourse in and around the public house. This four-volume reset edition presents a wide-ranging collection of primary sources which uncover the language and behaviour of local and state authorities, of peasants and town-dwellers, and of drinking companions and irate wives. The documents are translated and set in their social and historical context, providing a multidisciplinary collection that will be of great importance to scholars of all areas of social and cultural history of the early modern period. The vast majority of this material is published here for the first time, ensuring that the collection will open up new avenues of research. Volume 1 draws heavily from the Parisian police archives and includes inspectors’ reports, complaints by the general public and details of court cases to build a picture of drinking in early modern France. Volumes 2 and 3 address public drinking in the Holy Roman Empire through a variety of chronicles, civic ordinances, court records, travel reports and surveys of public houses. Volume 4 locates taverns within a broader analysis of America’s public houses, drawing on visual material as well as journal entries, business reports and newspaper articles. Each volume is accompanied by editorial introductions and is annotated to provide readers with a high-quality resource of scholarly material.

Multiple meningiomas: clinical, radiological, surgical, and pathological findings with outcome in four cats

The present report describes the clinical signs, magnetic resonance imaging (MRI) findings, surgical procedure, pathological findings and follow-up in four cats with multiple meningiomas; three castrated male and one spayed female domestic shorthair indoor cats, ranging in age from 11 to 14 years. In three of four cats, clinical signs at presentation were suggestive of a focal lesion. Three cats had two meningiomas and one had four meningiomas. Most of the tumours were supratentorial, one arose from the tentorium and one was infratentorial. The duration of presenting signs before surgery ranged from 10 days to 11 months. Postoperative MRI revealed complete gross tumour removal in three cases. In one cat with two cranial fossa meningiomas, subtotal excision with a small basal remnant (2 x 2 mm) of the ventral part of one meningioma lying on the floor of the skull, was observed. Based on histopathological architecture, six tumours revealed features of a transitional subtype meningioma, and four of a meningotheliomatous meningioma. In each cat, the multiple meningiomas were all assigned to the same histopathological group. The preoperative presenting signs had resolved by the follow-up examinations 4 weeks after surgery in two cats. Long-term follow-up evaluation revealed that surgically-induced or exacerbated neurological deficits in two cats had completely or almost completely resolved within 8 weeks of surgery. All patients are still alive 12 to 21 months after surgery and no clinical signs of recurrence could be detected at that time.

Oral mucosal lesions diagnosed in a stomatology service. An examination of clinico-pathological findings from the year 2003

During 2003, a total of 258 new patients with oral soft tissue lesions were admitted at the Stomatology Service of the Department of Oral Surgery and Stomatology at the University of Berne. For the present study, 185 patients with clinically and histopathologically verified diagnoses were included. The following data was collected: prevalence of oral mucosal lesions, distribution of benign, precancerous and malign lesions in different age groups, and the concordance of the referral with the working diagnosis at the Stomatology Service. The most frequent pathological soft tissue findings were fibrous hyperplasias (n = 44) and oral lichen planus (n = 30). Precancerous lesions were present in 41 cases (30 patients with oral lichen planus, eleven oral leukoplakias), and ten patients had oral malignomas. Most lesions were found in patients between the age of 40 and 60 years. The referral diagnosis concurred in 36.6% (n = 67) of the cases with the definite diagnosis before initiation of treatment, the working diagnosis in 70% (n = 128) of the cases. Therefore, it can be concluded that a specialised Stomatology Service serves as a center of competence due to large numbers of patients/cases seen and treated, and the resulting high level of clinical experience of the staff. Moreover, it is important in the primary diagnosis of oral squamous cell carcinoma, in collaboration with the referring dentist in private practice.

Proteolytic processing of chemokines: implications in physiological and pathological conditions

Chemokines are small, secreted proteins that orchestrate the migration of cells, which are involved in immune defence, immune surveillance and haematopoiesis. However, chemokines are also implicated in the pathology of various inflammatory diseases, cancers and HIV. The chemokine system is considerably large and has a redundancy in the repertoire of its inflammatory mediators. Therefore, strict regulation of chemokine activity is crucial. Chemokines are the substrate for various proteases including the serine protease CD26/dipeptidyl-peptidase IV and matrix metalloproteinases. Regulation by proteolytic cleavage controls and fine-tunes chemokine function by either enhancing or reducing its chemotactic activity or receptor selectivity. Often chemokines and the proteases that regulate them are produced in the same microenvironment and expression of both may be simultaneously induced by a common stimulus enabling the rapid regulation of chemokine activity. The overall impact of cleaved chemokines in cellular responses is very complex. In this review, we will give an overview on chemokine modification and the respective chemokine modifying proteases. Furthermore, we will summarize the emerging literature describing the consequences in inflammation, haematopoiesis, cancer and HIV infection upon proteolytic chemokine processing.

Traumatic extra-axial hemorrhage: correlation of postmortem MSCT, MRI, and forensic-pathological findings

PURPOSE: To evaluate the diagnostic accuracy of in situ postmortem multislice computed tomography (MSCT) and magnetic resonance imaging (MRI) in the detection of primary traumatic extra-axial hemorrhage. MATERIALS AND METHODS: Thirty forensic neurotrauma cases and 10 nontraumatic controls who underwent both in situ postmortem cranial MSCT and MR imaging before autopsy were retrospectively reviewed. Both imaging modalities were analyzed in view of their accuracy, sensitivity, and specificity concerning the detection of extra-axial hemorrhage. Statistical significance was calculated using the McNemar test. kappa values for interobserver agreement were calculated for extra-axial hemorrhage types and to quantify the agreement between both modalities as well as MRI, CT, and forensics, respectively. RESULTS: Analysis of the detection of hemorrhagic localizations showed an accuracy, sensitivity, and specificity of 89%, 82%, and 92% using CT, and 90%, 83%, and 94% using MRI, respectively. MRI was more sensitive than CT in the detection of subarachnoid hemorrhagic localizations (P = 0.001), whereas no significant difference resulted from the detection of epidural and subdural hemorrhagic findings (P = 0.248 and P = 0.104, respectively). Interobserver agreement for all extra-axial hemorrhage types was substantial (CT kappa = 0.76; MRI kappa = 0.77). The agreement of both modalitites was almost perfect (readers 1 and 2 kappa = 0.88). CONCLUSION: CT and MRI are of comparable potential as forensic diagnostic tools for traumatic extra-axial hemorrhage. Not only of forensic, but also of clinical interest is the observation that most thin blood layers escape the radiological evaluation.