Vascular anomalies (VAs) result from the defective development of the embryonic vascular system and feature dysplastic malformed vessels, which are not always apparent at birth. They do not regress over the patient's lifetime; they usually have commensurate growth during childhood and may worsen over time if not treated. VAs may cause chronic painful swelling, bleeding, functional deficits or vital structure obstruction. These patients' quality of life is usually impaired because of the chronicity and recurrence of the disease. We report on six cases of complicated VAs, refractory to current treatments, treated with rapamycin, an mTor inhibitor recently used in VAs. (C) 2017 Elsevier Masson SAS. All rights reserved.

Palpable thyroid nodules are present clinically in 4-7% of the population and their prevalence increases to 50%-67% when using high-resolution neck ultrasonography. By contrast, thyroid carcinoma (TC) represents only 5-20% of these nodules, which underlines the need for an appropriate approach to avoid unnecessary surgery. Frozen section (PS) has been used for more than 40 years in thyroid surgery to establish the diagnosis of malignancy. However, a controversy persists regarding the accuracy of FS and its place in thyroid pathology has changed with the emergence of fine-needle aspiration (FNA). A PubMed Medline and SpringerLink search was made covering the period from January 2000 to June 2012 to assess the accuracy of ES, its limitations and indications for the diagnosis of thyroid nodules. Twenty publications encompassing 8.567 subjects were included in our study. The average value of TC among thyroid nodules in analyzed studies was 15.5 %. ES ability to detect cancer expressed by its sensitivity (Ss) was 67.5 %. More than two thirds of the authors considered PS useful exclusively in the presence of doubtful ENA and for guiding the surgical extension in cases confirmed as malignant by FNA; however, only 33% accepted FS as a routine examination for the management of thyroid nodules. The influence of FS on surgical reintervention rate in nodular thyroid pathology was considered to be negligible by most studies, whereas 31 % of the authors thought that FS has a favorable benefit by decreasing the number of surgical re-interventions. In conclusion, the role of FS in thyroid pathology evolved from a mandatory component for thyroid surgery to an optional examination after a pre-operative FNA cytology. The accuracy of FS seems to provide no sufficient additional benefit and most experts support its use only in the presence of equivocal or suspicious cytological features, for guiding the surgical extension in cases confirmed as malignant by FNA and for the identification of other potentially confusing intraoperative findings.

Paravertebral malignant tumors constitute 4.8% of cancer cases in pediatric oncology and are mostly composed of neuroblastoma (46.4%) and soft tissue sarcomas (35.7%). We describe the case of a Caucasian 6-year-old boy who was admitted for middle back pain radiated to limbs and progressively increasing weakness of the legs, suggesting a spinal cord disease. The exploration revealed two paravertebral masses extending through the neural foraminae into the epidural space. The association with elevated serum neuron specific enolase suggested at first the diagnosis of neuroblastoma, but the pathological examination revealed a Burkitt's lymphoma. This is a rare location of sporadic Burkitt's lymphoma with neurologic syndrome as first symptoms.

The breast pathology includes a large array of entities for which macroscopic and microscopic analysis remains fundamental. Tissue and cell morphology allows in most cases the distinction between benign or malignant tumours and therefore provides the clinicians with essential information for the therapeutic strategy. In the Pathology laboratory, immunohistochemistry and molecular biology have improved the specificity of the diagnosis and have introduced new prognostic and predictive markers for tumour management. The last edition of the WHO classification, released in 2003, distinguishes 21 varieties of invasive carcinoma and 2 categories of intraepithelial neoplasia based on the morphology and immunohistochemical profile. Other diseases can affect the breast, although much less frequently, such as Paget's disease of the nipple, phyllode tumours, sarcomas, lymphomas... These diseases will not be reviewed here.

Bone marrow aplasias are characterized by a peripheral blood cells deficit resulting from a deficiency in the production of one or several blood cells lineages in bone marrow. Some of these aplasias are able to regenerate, others don’t respond to treatment. In most of the cases, spontaneously regenerating bone marrows are characterized by the presence of a great number of fibroblastic cells presenting a strong membranous alkaline phosphatase activity (ALP), this activity has completely disappeared in irreversible bone marrow aplasias. The aim of our work is to analyze phenotypic and functional characteristics of these ALP positive reticular cells. In this aim, we used a murine radio-induced aplasia model: a 4 Gy gamma rays total body irradiation induced an important but reversible bone marrow aplasia.

The study of the human hematopoietic system would be facilitated by availability of a relevant animal model. Because the medullar microenvironment is made of different types of cells, interactions between hematopoietic cells and stromal cells are difficult to analyze in detail. As an approach for establishing an in vivo model to dissect these interactions, we grafted murine bone marrow fibroblastic cells (MS‐5 cell line) with hematopoietic cells into the kidney capsule of syngenic mice. To identify the origin of cells present in the graft, we used green fluorescent protein–stable transfected MS‐5 cells for the transplantation. To analyze the evolution of stromal cells and identify hematopoietic cells able to develop in these conditions, we performed morphology, histochemistry, and immunohistology on tissue sections at different times after transplantation. When injected alone, MS‐5 cells differentiate into adipocytes. When injected with a bone marrow suspension or with isolated CD45+ cells (leukocytes), the stromal cells keep their fibroblastic morphology and their alkaline phosphatase expression and sustain granulopoiesis. When injected with hematopoietic stem cells called c‐kit+Sca‐1+Lin− suspension, clusters of hematopoietic cells are also observed: They do not present any granulopoietic activity and do not belong to B or T population nor to erythroid lineage. They are quiescent, induce bone marrow recovery and survival of lethally irradiated recipients, are able to form macroscopic colonies in the spleen, and are able to form very few colonies in vitro, suggesting that they are hematopoietic stem cells. In conclusion, our results show that reticular fibroblastic stromal cells MS‐5 sustain the survival of stem cells and are not able to induce their differentiation. However, they can control differentiation, proliferation, and/or survival of hematopoietic cells engaged in myeloid lineage.

Attempts were made to establish models to study interactions between marrow stromal cells and hematopoietic cells in vivo. The approach was to create a NOD-SCID-hu murine model of long-term human hematopoiesis by implantation of a human adult bone fragment. Nine to 12 weeks posttransplantation, human CD45+ cells were detected in the blood and the spleen of some mice. The histology of the human transplant showed that human bone fragment was viable at 9 weeks. Moreover, vessels of human origin, as assessed by immunohistochemical detection of human beta2-microglobulin, were observed in the mouse tissue surrounding the transplanted human fragment.

Purpose: The identification of fibroblast-like cells of the marrow stroma by means of alkaline phosphatase (ALP) cytochemistry reveals delicate ALP-positive structures interspersed among haematopoietic cells and arranged in a loosely meshed network. These cells are often referred to as 'reticular' cells and the network they form is known as the 'ALP network'. The purpose was to analyse the evolution of this ALP network in relation to haemopoietic regeneration after whole-body irradiation.Materials and methods: The total surface occupied by ALP-positive processes revealed by means of ALP cytochemistry was expressed as a ratio of the total marrow area. ALP-positive cells were counted using nuclei as the defining unit. Cell proliferation was analysed by the detection of bromodeoxyuridine (BrdU) incorporation. Fat cells were identified by oil red O staining and alpha-glycerophosphate dehydrogenase (alpha-GPDH) activity.Results: The ALP network and ALP-positive cell number began to increase 24 h after 4-Gy irradiation to reach a maximum after 72 h, when the bone marrow was almost completely empty of haemopoietic cells. This increase was in advance of haemopoietic recovery and was not due to cell proliferation. A decrease in the ALP network occurred in parallel with an increase in haemopoiesis and was accompanied by a transient increase in fat cells on day 7.Conclusions: These data indicate that the recovery of the ALP network, which is partially due to the recruitment of ALP-positive cells, occurs in advance of the haemopoietic recovery and that the equilibrium between fat cells and ALP-positive cells seems to be controlled by haemopoietic cells.

A 51-year old patient consults for abdominal swelling and persistent constipation. Clinical exploration shows the presence of a left iliac fossa tumor corresponding to a papillary serous adenocarcinoma of the fallopian tube after macroscopic and microscopic examination. The diagnostic and therapeutic problems caused by this rare gynecologic tumor are discussed.

s of the papers presented during the meeting of January 26, 2002, Université Libre de Bruxelles, Hôpital Erasme 95 ABSTRACTS OF THE PAPERS PRESENTED DURING THE MEETING OF JANUARY 26, 2002, UNIVERSITÉ LIBRE DE BRUXELLES, HÔPITAL ERASME Acta Clinica Belgica, 2002; 57-2 BOLD CONTRAST WITHIN TUMORS : HOW DOES IT CORRELATE TO PO 2 ? SIMULTANEOUS AND CO-LOCALIZED MEASUREMENT USING GRADIENTECHO FMRI AND OXYLITE-OXYFLO Christine Baudelet and Bernard Gallez Laboratory of Biomedical Magnetic Resonance and Laboratory of Medicinal Chemistry and Radiopharmacy, Université Catholique de Louvain, B-1200 Brussels, Belgium. BOLD imaging is often proposed as a tool to evaluate the responsiveness of tumors to oxygen changes. However, the signal is combination of flow and oxygen dependant parameters. Up to-now, the validity of the approach relies only on the comparison of BOLD signal with oxygen measurements carried out on different animals or animals measured at different times and location. We have developed an experimental setup which allows a simultaneous and co-localized measurement of pO2/flow and gradient echo fMRI. Isoflurane anesthetized mice were imaged at 4.7 Tesla. The slice for imaging was carefully selected such that it was positioned on the extremity of the Oxylite-/Oxyflo probe inserted into the intramuscularly implanted tumor. The spatial resolution for imaging was chosen to get voxels dimensions compatible with the sensitive area of the probe, while keeping a good BOLD contrast. GE images (200/20/45°) were continuously acquired during modification of gas breathing (air-50%O2-air-100%O2-air-Carbogen-air). A set of multi-echo GE images (200/ 6.1/6/45°) were then also acquired following the same protocol. From Multi-echo images, both T2* map and S 0 map were created. We computed (pixel-by-pixel analysis) the correlation of the three time series with the signals coming from OxyLite and OxyFlo. A typical co-localized and simultaneous recording of the GE signal and tumor pO 2 : EXPRESSION OF VEGF AND ITS RECEPTORS IN HUMAN ILIAC CREST AND FEMORAL BONE MARROW Z. Belaid, A. Thiry, A. Rodriguez, J. Boniver, Ch.M.Lapière, BV. Nusgens and MP. Defresne, Departement of Histology and Anatomy pathology, Departement of Orthopedics Surgery, Laboratory of Connectives Tissues Biology, University of liege, Sart-Tilman, Belgium The understanding of the mechanisms regulating hematopoiesis in normal bone marrow is required for progressing in the elucidation of the alteration occurring in many hematological malignancies such as myelodysplastic syndromes and leukemias. The aim of this study is to further explore the hypothesis that normal human hematopoiesis may be regulated by paracrine regulatory loops and by cross-talk between hematopoietic cells and their environment. The vascular network is the first candidate, which can be implicated, in these regulatory loops. We investigated by RT-PCR the mRNA level of the vascular endothelial cell growth factor (VEGF), its splice variants and its receptors, the VEGR-1/Flt1, the VEGFR-2/KDR and the specific VEGF165 isoform co-receptor, the neuropilin-1 (NRP-1) as well as its ligand semaphorin-III (SEM-III) known to function as VEGF antagonist. Bone marrows were collected from the iliac crest and the femur from 11 donors. VEGF121 and 165 are the main isoforms expressed at variable levels in iliac crest and femur while the extracellular matrix binding-VEGF189 is present in all samples, although at low level. Neither the 145 nor the 206 variants are detected. The VEGFR-2 is similarly expressed in all samples. Interestingly we observed a higher steady state level of VEGFR-1, NRP-1 and SEM-III in the femoral bone marrow, rich in adipocytes than in the hematopoietic iliac crest. To assess the cellular origin of this overexpression in femoral bone marrow, adipocytes were isolated from collagenase-digested femoral biopsies on the basis of their floating properties. The purified adipocyte population effectively expressed more NRP-1 mRNA than the sedimented cells population (stromal cells and hematopoietic cells). These results will be confirmed by immunostaining and western-blot analysis at the protein level. In conclusion, this preliminary study showed that the expression of NRP-1 in adipocytes is high when hematopoiesis is low, suggesting that these cells may play an active role in regulation of hematopoiesis and angiogenesis. In conclusion, BOLD contrast imaging is a powerful tool to appreciate the efficacy of a treatment of O 2 breathing. In carbogen breathing protocols, the variation in signal intensity is higher than using the oxygen protocol; this effect is likely to be due to the combination of an inflow effect together (increase in So) with a higher increase in pO 2 (increase in T2*). BOLD MRI is a non invasive method that allows a mapping of the response, and the variations observed are directly related to pO 2 changes. However, this technique has no value to estimate the basal pO 2 before treatment and BOLD MRI should be considered as a technique complementary to other methods allowing absolute quantification of the pO 2 . QUANTIFICATION OF MICROMETASTATIC DISEASE IN BREAST CANCER PATIENTS : OPTIMALISATION OF THE METHOD USING REAL-TIME QUANTITATIVE PCR FOR THE DETECTION OF CK19 MRNA IN BLOOD I. Benoy, H. Elst, P. Van Dam, P. Huget, S. Scharpé, E. Van Marck, L. Dirix, Oncology Center, AZ Sint-Augustinus, Antwerp, Belgium Department of Pathology, University Hospital Antwerp, Belgium Department of Clinical Biochemistry, University of Antwerp, Belgium Introduction : Earlier diagnosis has led to stage migration and better prognosis, but a subgroup of patients with early breast cancer continue to succumb to metastatic disease. Nodal status remains the most valuable marker to predict for the presence of occult spread. Morphological and molecular techniques suffer from low sensitivity and a lack of quantification. We have developed a sensitive and quantitative assay to detect breast cancer cells in venous blood using a quantitative RT-PCR identifying transcripts of the CK19 gene Methods : Total RNA was isolated from full blood samples of patients and healtzy volunteers using the Qiagen Rneasy ® total RNA kit. For generation of first strand cDNA, 2μg of total RNA was reverse transcripted with the high-Capacity cDNA Archive Kit (Applied Biosystems). A cDNA-specific CK19 Taqman TM primer and probe set was developed using Primer Express ® software. PCR was performed on the ABI Prism 7700 system and the standard thermal cycling protocol. The cycle at witch the emission intensity rises above baseline is referred to as cycle

s of the papers presented during the meeting of January 26, 2002, Universite Libre de Bruxelles, Hopital Erasme 95 ABSTRACTS OF THE PAPERS PRESENTED DURING THE MEETING OF JANUARY 26, 2002, UNIVERSITE LIBRE DE BRUXELLES, HOPITAL ERASME Acta Clinica Belgica, 2002; 57-2 BOLD CONTRAST WITHIN TUMORS : HOW DOES IT CORRELATE TO PO 2 ? SIMULTANEOUS AND CO-LOCALIZED MEASUREMENT USING GRADIENTECHO FMRI AND OXYLITE-OXYFLO Christine Baudelet and Bernard Gallez Laboratory of Biomedical Magnetic Resonance and Laboratory of Medicinal Chemistry and Radiopharmacy, Universite Catholique de Louvain, B-1200 Brussels, Belgium. BOLD imaging is often proposed as a tool to evaluate the responsiveness of tumors to oxygen changes. However, the signal is combination of flow and oxygen dependant parameters. Up to-now, the validity of the approach relies only on the comparison of BOLD signal with oxygen measurements carried out on different animals or animals measured at different times and location. We have developed an experimental setup which allows a simultaneous and co-localized measurement of pO2/flow and gradient echo fMRI. Isoflurane anesthetized mice were imaged at 4.7 Tesla. The slice for imaging was carefully selected such that it was positioned on the extremity of the Oxylite-/Oxyflo probe inserted into the intramuscularly implanted tumor. The spatial resolution for imaging was chosen to get voxels dimensions compatible with the sensitive area of the probe, while keeping a good BOLD contrast. GE images (200/20/45°) were continuously acquired during modification of gas breathing (air-50%O2-air-100%O2-air-Carbogen-air). A set of multi-echo GE images (200/ 6.1/6/45°) were then also acquired following the same protocol. From Multi-echo images, both T2* map and S 0 map were created. We computed (pixel-by-pixel analysis) the correlation of the three time series with the signals coming from OxyLite and OxyFlo. A typical co-localized and simultaneous recording of the GE signal and tumor pO 2 : EXPRESSION OF VEGF AND ITS RECEPTORS IN HUMAN ILIAC CREST AND FEMORAL BONE MARROW Z. Belaid, A. Thiry, A. Rodriguez, J. Boniver, Ch.M.Lapiere, BV. Nusgens and MP. Defresne, Departement of Histology and Anatomy pathology, Departement of Orthopedics Surgery, Laboratory of Connectives Tissues Biology, University of liege, Sart-Tilman, Belgium The understanding of the mechanisms regulating hematopoiesis in normal bone marrow is required for progressing in the elucidation of the alteration occurring in many hematological malignancies such as myelodysplastic syndromes and leukemias. The aim of this study is to further explore the hypothesis that normal human hematopoiesis may be regulated by paracrine regulatory loops and by cross-talk between hematopoietic cells and their environment. The vascular network is the first candidate, which can be implicated, in these regulatory loops. We investigated by RT-PCR the mRNA level of the vascular endothelial cell growth factor (VEGF), its splice variants and its receptors, the VEGR-1/Flt1, the VEGFR-2/KDR and the specific VEGF165 isoform co-receptor, the neuropilin-1 (NRP-1) as well as its ligand semaphorin-III (SEM-III) known to function as VEGF antagonist. Bone marrows were collected from the iliac crest and the femur from 11 donors. VEGF121 and 165 are the main isoforms expressed at variable levels in iliac crest and femur while the extracellular matrix binding-VEGF189 is present in all samples, although at low level. Neither the 145 nor the 206 variants are detected. The VEGFR-2 is similarly expressed in all samples. Interestingly we observed a higher steady state level of VEGFR-1, NRP-1 and SEM-III in the femoral bone marrow, rich in adipocytes than in the hematopoietic iliac crest. To assess the cellular origin of this overexpression in femoral bone marrow, adipocytes were isolated from collagenase-digested femoral biopsies on the basis of their floating properties. The purified adipocyte population effectively expressed more NRP-1 mRNA than the sedimented cells population (stromal cells and hematopoietic cells). These results will be confirmed by immunostaining and western-blot analysis at the protein level. In conclusion, this preliminary study showed that the expression of NRP-1 in adipocytes is high when hematopoiesis is low, suggesting that these cells may play an active role in regulation of hematopoiesis and angiogenesis. In conclusion, BOLD contrast imaging is a powerful tool to appreciate the efficacy of a treatment of O 2 breathing. In carbogen breathing protocols, the variation in signal intensity is higher than using the oxygen protocol; this effect is likely to be due to the combination of an inflow effect together (increase in So) with a higher increase in pO 2 (increase in T2*). BOLD MRI is a non invasive method that allows a mapping of the response, and the variations observed are directly related to pO 2 changes. However, this technique has no value to estimate the basal pO 2 before treatment and BOLD MRI should be considered as a technique complementary to other methods allowing absolute quantification of the pO 2 . QUANTIFICATION OF MICROMETASTATIC DISEASE IN BREAST CANCER PATIENTS : OPTIMALISATION OF THE METHOD USING REAL-TIME QUANTITATIVE PCR FOR THE DETECTION OF CK19 MRNA IN BLOOD I. Benoy, H. Elst, P. Van Dam, P. Huget, S. Scharpe, E. Van Marck, L. Dirix, Oncology Center, AZ Sint-Augustinus, Antwerp, Belgium Department of Pathology, University Hospital Antwerp, Belgium Department of Clinical Biochemistry, University of Antwerp, Belgium Introduction : Earlier diagnosis has led to stage migration and better prognosis, but a subgroup of patients with early breast cancer continue to succumb to metastatic disease. Nodal status remains the most valuable marker to predict for the presence of occult spread. Morphological and molecular techniques suffer from low sensitivity and a lack of quantification. We have developed a sensitive and quantitative assay to detect breast cancer cells in venous blood using a quantitative RT-PCR identifying transcripts of the CK19 gene Methods : Total RNA was isolated from full blood samples of patients and healtzy volunteers using the Qiagen Rneasy ® total RNA kit. For generation of first strand cDNA, 2μg of total RNA was reverse transcripted with the high-Capacity cDNA Archive Kit (Applied Biosystems). A cDNA-specific CK19 Taqman TM primer and probe set was developed using Primer Express ® software. PCR was performed on the ABI Prism 7700 system and the standard thermal cycling protocol. The cycle at witch the emission intensity rises above baseline is referred to as cycle

Recently, Panoutsakopoulos et al. (1999) reported 2 cases of aneurysmal bone cysts with a recurrent (16;17)(q22;p13) translocation. We present here two additional cases harboring the same translocation as well as additional chromosomal changes.

Invasive aspergillosis is a mycelial infection, associated in 15 to 30% of cases with a malignant hemopathy or with other types of cancers. It also constitutes a complication induced by high dosage corticotherapy or long term antibiotherapy. On the occasion of an autopsy of invasive aspergillosis, we review the anatomo-clinical entities associated with aspergillus, the etiopathogenic factors and the diagnostic difficulties.

Translocation t(2p;3q) is a rare but recurrent finding in myeloid disorders. We present the first case of primary myelofibrosis with t(2;3)(p21;q26) as the sole chromosomal anomaly. The comparison with the 11 other previously published myeloid-associated t(2p;3q) cases confirms that this nonrandom translocation involves a pluripotent stem cell and is associated with a poor prognosis.

We report a case of Kikuchi‐Fujimoto disease with cutaneous involvement in a European man. In contrast, the disease is most prevalent in women of Asian descent. The condition is probably under‐recognized when histology is not used to identify lymphadenitis of protracted course. Skin lesions may mimic clinically other unrelated disorders including lymphomas and immune or infectious dermatoses. Histology of skin lesions may bring a clue to the diagnosis when apoptotic plasmacytoid monocytes are recognized. The relationship between Kikuchi‐Fujimoto disease and lupus erythematous remains a matter of debate.

UNLABELLED:Leishmaniasis refers to a spectrum of diseases caused by Leishmania. Clinically, three types of leishmaniasis can be distinguished: the cutaneous, mucous and visceral leishmaniasis, the latter being caused by Leishmania donovani. CASE REPORT:An 11-year-old Thai, living in Belgium for 6 years, had surgery for a vertebral osteosarcoma with pulmonary metastases, followed by polychemotherapy, then pulmonary metastasectomy. During a post-chemotherapy bone marrow aplasia, febrile episode with a general condition impairment was noted and first treated by broad-spectrum antibiotherapy, then by amphotericin B, in the absence of any accurate etiology. The outcome first was favorable. Nevertheless, 7 months later, the visceral leishmaniasis diagnosis was made because of the recurrence of the same symptoms. Classical treatments by antimony derivatives (Glucantim), then liposomal amphotericin (Ambisome) proved to be inefficient. A liposomal amphotericin-gamma interferon association suppressed the symptoms without eradicating the parasite. The patient was given a maintenance therapy based on liposomal amphotericin. CONCLUSION:The stubborn and recurring nature of this chronic visceral leismaniosis can be due to the immune deficit inherent in the polychemotherapy performed in order to treat the metastatic osteosarcoma which currently is in first full remission.