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Becheanu G.,Carol Davila University of Medicine and Pharmacy | Becheanu G.,Victor Babes Institute of Pathology | Pop A.,Fundeni Clinical Institute | Dumbrava M.,Fundeni Clinical Institute | And 4 more authors.
Archives of the Balkan Medical Union | Year: 2013

Background: Whipple's disease is a rare chronic systemic disease caused by Tropheryma whipplei, an ubiquitous Gram-positive Actinobacteria. The estimated incidence of the disease is less than 1 per 1 milion, males predominantly affected. Methods: We report five new cases of Whipple's disease, four males and one female, diagnosed in our clinic from 2002 to 2012. Diagnosis was reached with the help of the upper digestive endoscopy and subsequent histopathological examination of the small bowel biopsy specimen and lymph node biopsy (one case) and by electron microscopy. The main symptoms were arthralgia, weight loss and diarrhea. Results: The endoscopic feature of the small bowel mucosa varied from congestion, granularity of the mucosa to whitish plaques. All patients showed PAS positive, Ziehl-Neelsen negative macrophages identified in the lamina propria of the small bowel mucosa and, in one case, inside of a retroperitoneal lymph node. The diagnosis was confirmed by electron microscopy in all cases. Classic Whipple's disease was the diagnosis in all five cases, but involvement of the endocardium in one case and lymph nodes in two cases was observed. The clinical evolution was favourable after antibiotic treatment in four patients, but one was lost to follow-up. In two patients, the duodenal biopsies performed six months after treatment showed a reduced number of PAS positive macrophages in the lamina propria, in the absence of any clinical symptoms. Conclusions; Tropheryma whipplei infection may induce a disease with many possible clinical features. In cases with extragastrointestinal symptoms a high level of clinical suspicion is necessary in order to reach the diagnosis. Copyright © 2013 CELSIUS.

Curici A.,University of Bucharest | Codrici E.,Victor Babes Institute of Pathology | Mihai S.,Victor Babes Institute of Pathology | Tanase C.P.,Victor Babes Institute of Pathology | And 5 more authors.
Romanian Journal of Neurology/ Revista Romana de Neurologie | Year: 2014

Objectives. Muscle contusion is the most common form of muscle injury and the muscle endogenous regenerative capacity can compensate for non-extensive damage. However, severe traumatic injuries may overcome this capacity. More effective therapeutic strategies are still needed, so the exploration of the molecular context during muscle regeneration might provide new insights.Materials and methods. We investigated the expression pattern of a reputed angiogenic molecule, CD105/endoglin, by measuring the tissue and serum concentration by multiplex assay in normal and Dmdmdx dystrophic mice by following the distribution of the cellular sources by immunofluorescence in a mouse model of acute muscle contusion.Results. Maximal tissue concentration in normal animals was obtained 48h post-injury, and then started to decline, with only one other significantly increased value 5 days after injury. In dystrophic mice, tissue levels were globally much higher than in normal animals and started rising 1h post-injury and were maintained elevated all along the regenerations process. In situ immunolabelling highlighted the increased positive population mostly in the inflammatory areas. Double staining separates distinct subsets based on hematopoietic marker CD45 and the endothelial marker CD31 co-expression in endoglin positive cells.Conclusions. This study offers a timeline of endoglin expression during normal and pathologic muscle regeneration, providing evidence that the major wave corresponds to the inflammatory stage of muscle regeneration, deriving from multiple cellular sources such as endothelial cells, blood cells and also other interstitial cells that become activated during this process. © 2014 Editura Medicala AMALTEA. All Rights reserved.

Ceafalan L.C.,Carol Davila University of Medicine and Pharmacy | Ceafalan L.C.,Victor Babes Institute of Pathology | Manole E.,Colentina Research Center | Manole E.,Victor Babes Institute of Pathology | And 6 more authors.
Romanian Journal of Neurology/ Revista Romana de Neurologie | Year: 2015

Objectives. Muscle regeneration after trauma is a complex phenomenon involving several cellular processes, such as angiogenesis, inflammation, fibrosis, activation of satellite cells and their differentiation into myocytes and myotubes. Although many studies explored these mechanisms in the last years, there is still an unmet need to find new therapy targets, especially regarding some cellular molecules involved in muscular recovery after mechanical or pathological injury. In the present study we investigated the dynamics of endothelin-1 (ET-1), an important factor that has been shown to be involved in all stages of tissue regeneration, but which is poorly investigated in skeletal muscle. Materials and Methods. We used an experimental animal model of acute mechanical trauma on mouse gastrocnemius muscle. ET-1 levels were investigated at different time-points after muscle injury by in situ immunofluorescence, xMAP assay on tissue and serum samples, and Western Blot analysis. Results. By xMAP assay, tissue ET-1 levels increased significantly up to the 5th day after trauma, correlated with serum levels. xMAP assay was confirmed by Western blot analysis which showed a significant increase in the level of ET-1 towards the end of the first week after trauma. This corresponds with the inflammatory stage of the regeneration process, followed by angiogenesis and satellite cell activation. In situ immunostaining showed a multiplication of interstitial cells expressing ET-1 in the first week after muscle injury. Two cellular subtypes were detected in the connective tissue - one is represented by blood-derived CD45 positive cells and the other by local interstitial cells. Such cells were detected in all connective tissue compartments, in close association with CD56 positive satellite cells, myoblasts and myotubes and most of them co-express sca-1. Conclusions. The present study demonstrated that ET-1 is synthesized mostly by mesenchymal progenitors and their number greatly increases after mechanical trauma in muscle interstitium. Based on ET-1 expression and their close association with activated satellite cells, such cells could have a paracrine influence not only over angiogenesis but also over fiber regeneration. ET-1 appears as an important molecule working in conjunction with other various signalling pathways especially during first stages of the regeneration process after acute mechanical injury. ET-1 and its receptors could become therapeutic targets, especially for inflammatory myopathies and muscular dystrophies with significant pathological fibrosis. © 2015, Editura Medicala AMALTEA. All rights reserved.

Ceafalan L.C.,Carol Davila University of Medicine and Pharmacy | Ceafalan L.C.,Victor Babes Institute of Pathology | Popescu B.O.,Victor Babes Institute of Pathology | Popescu B.O.,Carol Davila University of Medicine and Pharmacy | And 2 more authors.
BioMed Research International | Year: 2014

Skeletal muscle, a tissue endowed with remarkable endogenous regeneration potential, is still under focused experimental investigation mainly due to treatment potential for muscle trauma and muscular dystrophies. Resident satellite cells with stem cell features were enthusiastically described quite a long time ago, but activation of these cells is not yet controlled by any medical interventions. However, after thorough reports of their existence, survival, activation, and differentiation there are still many questions to be answered regarding the intimate mechanism of tissue regeneration. This review delivers an up-to-date inventory of the main known key players in skeletal muscle repair, revealed by various models of tissue injuries in mechanical trauma, toxic lesions, and muscular dystrophy. A better understanding of the spatial and temporal relationships between various cell populations, with different physical or paracrine interactions and phenotype changes induced by local or systemic signalling, might lead to a more efficient approach for future therapies. © 2014 Laura Cristina Ceafalan et al.

Ceafalan L.C.,Carol Davila University of Medicine and Pharmacy | Ceafalan L.C.,Victor Babes Institute of Pathology | Manole E.,Victor Babes Institute of Pathology | Tanase C.P.,Victor Babes Institute of Pathology | And 5 more authors.
Anatomical Record | Year: 2015

Angiogenesis is a key event during tissue regeneration, but the intimate mechanisms controlling this process are still largely unclear. Therefore, the cellular and molecular interplay along normal tissue regeneration should be carefully unveiled. To this matter, we investigated by xMAP assay the dynamics of some angiogenic factors known to be involved in tissue repair, such as follistatin (FST), Placental Growth Factor-2 (PLGF-2), epidermal growth factor (EGF), betacellulin (BTC), and amphiregulin (AREG) using an animal model that mimics acute muscle contusion injuries. In situ immunofluorescence was used for the evaluation and tissue distribution of their cellular sources. Tissue levels of explored factors increased significantly during degeneration and inflammatory stage of regeneration, peaking first week postinjury. However, except for PLGF-2 and EGF, their levels remained significantly elevated after the inflammatory process started to fade. Serum levels were significantly increased only after 24 h for AREG and EGF. Though, for all factors except FST, the levels in injured samples did not correlate with serum or contralateral tissue levels, excluding the systemic influence. We found significant correlations between the levels of EGF and AREG, BTC, FST and FST and AREG in injured samples. Interstitial cells expressing these factors were highlighted by in situ immunolabeling and their number correlated with measured levels dynamics. Our study provides evidence of a dynamic level variation along the regeneration process and a potential interplay between selected angiogenic factors. They are synthesized, at least partially, by cell populations residing in skeletal muscle interstitium during regeneration after acute muscle trauma. © 2015 Wiley Periodicals, Inc.

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