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Testosterone production by the fetal testis depends on a functional relationship between hCG and the LH/chorionic gonadotropin receptor (LHCGR). Failure of the receptor to correctly respond to its ligand leads to impaired sexual differentiation in males. A phenotypically female patient with pubertal delay had a 46,XY karyotype and was diagnosed with 46,XY disorder of sex development (DSD). Novel compound heterozygous LHCGR mutations were found in the signal peptide: a duplication p.L10_Q17dup of maternal origin, and a deletion (p.K12_L15del) and a p.L16Q missense mutation of paternal origin. cAMP production was very low for both the deletion and duplication mutations and was halved for the missense mutant. The duplication and missense mutations were both expressed intracellularly, but at very low levels at the cell membrane; they were most likely retained in the endoplasmic reticulum. The deletion mutant had a very limited intracellular expression, indicating impaired biosynthesis. There was reduced expression of all three mutants, which was most marked for the deletion mutation. There was also decreased protein expression of all three mutant receptors. In the deletion mutation, the presence of a lower-molecular-weight band corresponding to LHCGR monomer, probably due to lack of glycosylation, and a lack of bands corresponding to dimers/oligomers suggests absent ER entry. This novel case of 46,XY DSD illustrates how different LHCGR signal peptide mutations led to complete receptor inactivation by separate mechanisms. The study underlines the importance of specific regions of signal peptides and expands the spectrum of LHCGR mutations.

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. Les malformations vasculaires (MV) sont des anomalies de développement des vaisseaux pendant la vie intra-utérine ; elles sont présentes à la naissance mais parfois découvertes tardivement. Elles ne présentent aucune tendance à la régression spontanée, mais au contraire une évolution progressive émaillée de poussées inflammatoires tout au long de la vie. Selon le type de MV, les complications peuvent être esthétiques, douloureuses, fonctionnelles, hémorragiques ou systémiques. La qualité de vie des patients porteurs de MV peut être fortement affectée en raison des symptômes présentés ainsi que du caractère chronique, durable et récidivant de l’affection. Nous présentons l’évolution clinique, biologique et radiologique de 6 patients souffrant de MV complexes réfractaires aux traitements conventionnels qui ont bénéficié d’une nouvelle approche thérapeutique par rapamycine (Sirolimus®), un inhibiteur de la voie mTor (pour mammalian target of rapamycin).

Glycoprotein hormones are complex hormonally active macromolecules. Luteinizing hormone (LH) is essential for the postnatal development and maturation of the male gonad. Inactivating Luteinizing hormone beta (LHB) gene mutations are exceptionally rare and lead to hypogonadism that is particularly severe in males. We describe a family with selective LH deficiency and hypogonadism in two brothers. DNA sequencing of LHB was performed and the effects of genetic variants on hormone function and secretion were characterized by mutagenesis studies, confocal microscopy and functional assays. A 20-year-old male from a consanguineous family had pubertal delay, hypogonadism and undetectable LH. A homozygous c.118_120del (p.Lys40del) mutation was identified in the patient and his brother, who subsequently had the same phenotype. Treatment with hCG led to pubertal development, increased circulating testosterone and spermatogenesis. Experiments in HeLa cells revealed that the mutant LH is retained intracellularly and showed diffuse cytoplasmic distribution. The mutated LHB heterodimerizes with the common alpha-subunit and can activate its receptor. Deletion of flanking glutamic acid residues at positions 39 and 41 impair LH to a similar extent as deletion of Lys40. This region is functionally important across all heterodimeric glycoprotein hormones, because deletion of the corresponding residues in hCG, follicle-stimulating hormone and thyroid-stimulating hormone beta-subunits also led to intracellular hormone retention. This novel LHB mutation results in hypogonadism due to intracellular sequestration of the hormone and reveals a discrete region in the protein that is crucial for normal secretion of all human glycoprotein hormones.

A 10-year-old girl with a family history of Hodgkin's lymphoma presented with a 2 month history of cervical lymphadenopathy and weight loss. Biopsy indicated concomitant nodal involvement by Langerhans cell histiocytosis and Hodgkin's lymphoma. Such an association is rare, especially so in children, but is not an isolated phenomenon, thereby prompting the question of whether Langerhans cell histiocytosis is a reactive or a neoplastic process.

Aero-digestive tract squamous intra-epithelial neoplasia is a disease whose genetic and epigenetic features lead to clinical signs and well codified histologic features. This publication aims to review the molecular alterations which have been identified in these lesions, to clarify the clinical manifestations and to discuss the proposed histological classification.

Dear Editor, Pituitary gigantism is caused by chronic growth hormone (GH) hypersecretion by a pituitary lesion before epiphyseal fusion. Genetic causes have been identified in nearly 50% of patients with pituitary gigantism, with germline mutations in the AIP gene being the most frequent cause (Rostomyan et al. 2015). Recently, a new form of pituitary gigantism, X-linked acrogigantism (X-LAG), was described (Trivellin et al. 2014). X-LAG is due to chromosome Xq26.3 duplication and GPR101 is the disease-associated gene (Trivellin et al. 2014 ; Iacovazzo et al. 2016). X-LAG is characterized by of mixed GH/prolactin-secreting pituitary macroadenomas and/or hyperplasia in early childhood (Beckers et al. 2015). X-LAG typically occurs sporadically in females, but somatic mosaicism also occurs in males ; familial mother-to-son transmission of the Xq26.3 duplication has been reported in three familial isolated pituitary adenoma families (Trivellin et al. 2014 ; Daly et al. 2016 ; Gordon et al. 2016 ; Iacovazzo et al. 2016). The clinical presentation of X-LAG syndrome differs from other genetic forms of pituitary gigantism (Rostomyan et al. 2015) and many well-known historical cases of gigantism share the clinical characteristics of X-LAG syndrome (Beckers et al. 2015 ; Rostomyan et al. 2015). If untreated during childhood X-LAG leads to established extreme gigantism (>1.9 meters) before puberty (Daly et al. 2016)...

Craniopharyngioma is the most common pituitary tumor in childhood. It can compromise the pubertal development because of its evolution or treatment. Syndrome of Klinefelter is the most common cause of hipergonadotrophic hypogonadism in males. The concomitant presentation of both entities is extremely low (1/109) and the pathophysiological association is questionned. We present the case of a 18-year-old Belgian patient. He had a diagnosis of craniopharyngioma in childhood and he presented with panhypopituitarism after radiotherapy and surgical treatment. At the age of 14, he started pubertal induction with gonadotropin therapy without clinical response. Asociación de craneofaringioma y síndrome de Klinefelter en la transición puberal: un desafío diagnóstico Craniopharyngioma and Klinefelter syndrome during the pubertal transition: A diagnostic challenge A genetic evaluation confirmed a homogeneous 47, XXY karyotype. Failure of exogenous gonadotropin therapy revealed the hidden association of primary and secondary hypogonadism, demonstrating the importance of the followup and a multidisciplinary approach in these patients.Los craneofaringiomas son de los tumores hipofisarios más frecuentes en la niñez y, sea por su evolución o por el tratamiento que requieren, pueden comprometer el desarrollo puberal. El síndrome de Klinefelter es la causa más frecuente de hipogonadismo hipergonadotrópico en el varón. La presentación concomitante de ambas entidades es extremadamente baja (1/109) y plantea un interrogante acerca de una probable asociación fisiopatológica. Se presenta el caso de un paciente belga de 18 años, con diagnóstico de craneofaringioma en la niñez y panhipopituitarismo luego del tratamiento quirúrgico y radioterápico. Al llegar a los 14 años, se inició la inducción puberal con gonadotropinas. Ante la falta de respuesta clínica, se completó una evaluación genética, que evidenció, de manera homogénea, una trisomía XXY. La falta de respuesta al tratamiento de inducción con gonadotropina exógena reveló la asociación de hipogonadismo primario y secundario, que demostró la importancia del seguimiento multidisciplinario que estos pacientes requieren.

We recently showed that Xq26.3 microduplications cause X-linked acrogigantism (X-LAG). X-LAG patients mainly present with growth hormone and prolactin-secreting adenomas and share a minimal duplicated region containing at least four genes. GPR101 was the only gene highly expressed in their pituitary lesions, but little is known about its expression patterns. In this work, GPR101 transcripts were characterized in human tissues by 5′-Rapid Amplification of cDNA Ends (RACE) and RNAseq, while the putative promoter was bioinformatically predicted. We investigated GPR101 mRNA and protein expression by RT-quantitative PCR (qPCR), whole-mount in situ hybridization, and immunostaining, in human, rhesus monkey, rat and zebrafish. We identified four GPR101 isoforms characterized by different 5′-untranslated regions (UTRs) and a common 6.1kb long 3′UTR. GPR101 expression was very low or absent in almost all adult human tissues examined, except for specific brain regions. Strong GPR101 staining was observed in human fetal pituitary and during adolescence, whereas very weak/absent expression was detected during childhood and adult life. In contrast to humans, adult monkey and rat pituitaries expressed GPR101, but in different cell types. Gpr101 is expressed in the brain and pituitary during rat and zebrafish development; in rat pituitary, Gpr101 is expressed only after birth and shows sexual dimorphism. This study shows that different GPR101 transcripts exist and that the brain is the major site of GPR101 expression across different species, although divergent species- and temporal-specific expression patterns are evident. These findings suggest an important role for GPR101 in brain and pituitary development and likely reflect the very different growth, development and maturation patterns among species.

Breast cancer is rare in male patients. Certain predisposing factors, be they genetic (e.g., BRCA2 gene mutations) or hormonal (imbalance between estrogen and androgen levels), have been implicated in male breast cancer pathophysiology. Male-to-female (MtF) transsexualism is a condition that generally involves cross-sex hormone therapy. Anti-androgens and estrogens are used to mimic the female hormonal environment and induce the cross-sex secondary characteristics. In certain situations, the change in the hormonal milieu can be disadvantageous and favor the development of hormone-dependent pathologies, such as cancer. We report a case of a MtF transgender patient who developed breast cancer after 7 years of cross-sex hormonal therapy. The patient was found to be BRCA2 positive, and suffered recurrent disease. The patient was unaware of being a member of an established BRCA2 mutation-positive kindred. This represents the first case of a BRCA2 mutation predisposing to breast cancer in a MtF transgender patient.

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 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.

Haemangiomas are ubiquitous vascular tumours that rarely involve the pericardium. We report the case of a symptomatic patient with pericardial haemangioma reproducing the pathognomonic MR findings that have been describedfor haemangiomas of other locations.

A 10-year-old girl with a family history of Hodgkin's lymphoma presented with a 2 month history of cervical lymphadenopathy and weight loss. Biopsy indicated concomitant nodal involvement by Langerhans cell histiocytosis and Hodgkin's lymphoma. Such an association is rare, especially so in children, but is not an isolated phenomenon, thereby prompting the question of whether Langerhans cell histiocytosis is a reactive or a neoplastic process.

We performed this study in order to evaluate the diagnostic accuracy of whole-body fluorodeoxyglucose positron emission tomography (FDG PET) imaging and somatostatin receptor scintigraphy (SRS) for localizing primary carcinoid tumours and evaluating the extent of the disease. A secondary aim was to correlate those findings with the histological characteristics of the lesions. FDG PET was performed in 17 patients and SRS in 16. All patients had pathologically proven carcinoids. All lesions were verified by histopathological analysis or by follow-up. Ki-67 and p53 expression were assessed as an indicator of the tumours' aggressiveness. FDG PET correctly identified 4/7 primary tumours and 8/11 metastatic spreads, as compared to six and 10 respectively, for SRS. Most tumours were typical carcinoids with low Ki-67 expression. No correlation was found between the histological features and the tracer's uptake. We conclude that SRS remains the modality of choice for evaluating patients with carcinoid tumours, regardless of their proliferative activity. FDG PET should be reserved to patients with negative results on SRS.

ObjectiveThe LH/chorionic gonadotrophin receptor (LHCGR) is a G protein-coupled receptor (GPCR) that plays a central role in male sexual differentiation, regulation of ovarian follicular maturation, ovulation and maintenance of corpus luteum and pregnancy, as well as maintenance of testicular testosterone production. Mutations in theLHCGRgene are very rare. The aim of this work was to study the clinical and molecular characteristics of a rare familialLHCGRmutation.MethodsFive affected members of a family, including a phenotypically female, but genotypically male (46,XY), patient with Leydig cell hypoplasia type 1 and four genotypically female siblings with reproductive abnormalities, were studied genetically. Cell trafficking studies as well as signalling studies of mutated receptor were performed.ResultsThe five affected patients were all homozygous for a novel mutation in theLHCGRgene, a deletion of guanine in position 1850 (1850delG). This resulted in a frameshift affecting most of the C-terminal intracellular domain.In vitrostudies demonstrated that the 1850delG receptor was completely incapable of transit to the cell membrane, becoming trapped within the endoplasmic reticulum. This could not be rescued by small-molecule agonist treatment or stimulated intracellularly by co-expression of a yoked human chorionic gonadotrophin.ConclusionsThis novelLHCGRmutation leads to complete inactivation of the LHCGR receptor due to trafficking and signalling abnormalities, which improves our understanding of the impact of the affected structural domain on receptor trafficking and function.

McCune Albright syndrome (MAS) is a clinical association of endocrine and nonendocrine anomalies caused by postzygotic mutation of the GNAS1 gene, leading to somatic activation of the stimulatory α-subunit of G protein (Gsα). Important advances have been made recently in describing pathological characteristics of many MAS-affected tissues, particularly pituitary, testicular, and adrenal disease. Other rarer disease related features are emerging.The objective of the investigation was to study the pathological and genetic findings of MAS on a tissue-by-tissue basis in classically and nonclassically affected tissues.This was a comprehensive autopsy and genetic analysis.The study was conducted at a tertiary referral university hospital.An adult male patient with MAS and severe disease burden including gigantism was the subject of the study.Interventions included clinical, hormonal, and radiographic studies and gross and microscopic pathology analyses, conventional PCR, and droplet digital PCR analyses of affected and nonaffected tissues.Pathological findings and the presence of GNAS1 mutations were measured.The patient was diagnosed with MAS syndrome at 6 years of age based on the association of café-au-lait spots and radiological signs of polyostotic fibrous dysplasia. Gigantism developed and hyperprolactinemia, hypogonadotropic hypogonadism, and hyperparathyroidism were diagnosed throughout the adult period. The patient died at the age of 39 years from a pulmonary embolism. A detailed study revealed mosaiscism for the p.R201C GNAS1 mutation distributed across many endocrine and nonendocrine tissues. These genetically implicated tissues included rare or previously undescribed disease associations including primary hyperparathyroidism and hyperplasia of the thymus and endocrine pancreas.This comprehensive pathological study of a single patient highlights the complex clinical profile of MAS and illustrates important advances in understanding the characteristics of somatic GNAS1-related pathology across a wide range of affected organs.

Summary Heterozygous germline inactivating mutations in the aryl hydrocarbon receptor-interacting protein (AIP) gene lead to pituitary adenomas that most frequently present in the setting of familial isolated pituitary adenoma syndrome, usually as somatotropinomas and prolactinomas. More recently, they have been found in a significant percentage of young patients presenting with pituitary macroadenoma without any apparent family history. We describe the case of a 19-year-old man who presented with a gigantic somatotropinoma. His family history was negative. His peripheral DNA showed a heterozygous AIP mutation (p.I13N), while tumor tissue only had the mutated allele, showing loss of heterozygosity (LOH) and suggesting that the mutation caused the disease. Learning points AIP mutations may be observed in sporadic somatotrope adenomas occurring in young patients. LOH is a strong indicator that an AIP variant is disease causing. Somatotrope adenomas in carriers of AIP mutations are generally larger and more difficult to cure.

A 40-yr-old female presented with an extensive lesion of the sellar area and the sphenoid sinus, spreading to the optic nerves and associated with pachymeningitis. Histological findings were consistent with an inflammatory pseudotumor, and steroid treatment allowed the disappearance of all the lesions. Inflammatory pseudotumors of the pituitary are very rare. This case appears unique with regard to the extension of the lesions and the dramatic response to medical treatment. The differential diagnosis of inflammatory lesions of the pituitary is difficult. It relies mainly on histological analysis and includes sarcoidosis, Wegener's granulomatosis, histiocytosis (Langerhans, Rosai-Dorfman, and Erdheim-Chester diseases) and lymphocytic hypophysitis.

The routine use of arterial grafts in coronary surgery is facilitated by peroperative adjunction of antispasmodic drug to reduce the event of spasm. Diltiazem has been favoured in most clinical studies devoted to the radial artery graft. The aim of this study was to assess the efficacy of a spasm preventing protocol associating hydrostatic dilation of the graft with a diluted solution of papaverine and nicardipine infusion, starting preoperatively and continued postoperatively in i.v. and per os forms. Between September 1996 and March 1997, a consecutive series of 50 patients underwent myocardial revascularization using the radial artery. The radial artery was prepared by hydrostatic dilation with papaverine (1%) and nicardipine was administrated at 0.25 microgram/kg/min and titrated according to the arterial systemic pressure. Operative mortality was 4% (sepsis). There was no evidence of perioperative MI nor hypoperfusion syndrome. Mean CKMB level at 18 hours was 36 micrograms/l. No ischaemic anomalies of the ECG were detected. Angiography performed in the last 20 patients showed a 98% (51/52) permeability rate for all graft; 19/20 radial grafts (95%) were patent. One radial graft presented a 50% stenosis at the proximal anastomosis, and another a moderate spasm (40%) in the middle part of the conduit. This study confirms that the radial artery conduit can be used with satisfactory results for routine coronary artery bypass. The use of nicardipine allows the control the vasoreactivity of the radial graft without totally obviating at least angiographic spasm. This drug is easy to titrate, and well tolerated in association to beta-blockers in the routine perioperative management of the coronary patients.

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.

Hypertrophy and hyperplasia of pacinian corpuscles in the hand are very rare, with only 29 cases previously published. We describe two new cases in middle-aged patients whose chief complaint was progressive digital pain. Immunohistochemical and electronmicroscopic investigations indicated that the lump replicates the structure of enlarged pacinian corpuscles and should not be considered as a real tumor, nor connected to neurofibromatosis. No recurrence was observed after surgical excision. Although local trauma was encountered in 55% of the reported cases, the pathogenesis of such a lesion is still speculative.

Resume On observe trois types de leishmaniose : la leishmaniose cutanee, la leishmaniose muqueuse et la leishmaniose viscerale, cette derniere etant provoquee par Leishmania donovani. Observation. Un enfant de 11 ans, d'origine thailandaise, residant en Belgique depuis 6 ans, est opere d'un osteosarcome vertebral, metastase au niveau pulmonaire, ce qui conduit a une polychimiotherapie suivie d'une metastasectomie pulmonaire. Un episode d'hyperthermie, sans cause precise, avec alteration de l'etat general lors d'une aplasie medullaire post-chimiotherapique est traite par une antibiotherapie a large spectre puis par amphotericine b. L'evolution est d'abord favorable puis, 7 mois plus tard, un diagnostic de leishmaniose viscerale est pose a la suite de la reapparition des memes symptomes. Les traitements classiques par derives de l'antimoine (Glucantime®), puis par amphotericine liposomale (Ambisome®) se montrent inefficaces. Une association amphotericine liposomale et interferon gamma amende les symptomes sans eradiquer le parasite de l'organisme. Face a cette leishmaniose viscerale chronique, le patient recoit un traitement de maintenance par amphotericine liposomale. Conclusion. Le caractere refractaire et recidivant de cette leishmaniose viscerale chronique pourrait etre secondaire au deficit immunitaire inherent a la polychimiotherapie recue en raison d'un osteosarcome metastatique, actuellement en premiere remission complete.

OBJECTIVE: To examine the factors associated with liver steatosis in severely obese subjects and to test the potential reversibility of fatty liver after weight loss. DESIGN: Retrospective clinical study. SUBJECT: 528 obese patients before bariatric surgery and 69 obese subjects of the initial cohort evaluated before and 27±15 months after gastroplasty. MEASUREMENTS: Fatty deposition (scored as mild, moderate or severe) and inflammatory changes were evaluated in liver biopsies; clinical (body mass index (BMI), age, gender, duration of obesity) and biological (glucose, triglycerides, liver enzymes) parameters were related to histological findings. RESULTS: 74% of the 528 biopsies showed fatty change, estimated as mild in 41% of cases, moderate in 32% and severe in 27%. The prevalence of steatosis was significantly higher in men than in women (91% vs 70%, P=0.001) and in patients with impaired glucose tolerance or type 2 diabetes compared with nondiabetics (89% vs 69% P=0.001). The severity of the steatosis was associated with BMI (P=0.002) but not with the duration of obesity or the age of the patient. When compared with patients without fatty change, those with liver steatosis had significantly higher fasting plasma glucose (5.5 mmol/l vs 5.1 mmol/l, P=0.007) and triglycerides (1.8 mmol/l vs 1.3 mmol/l, P=0.002). Mean serum liver enzyme activities (alkaline phosphatase, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and gamma-glutamyl-transpeptidase (γGT) were significantly (P

Primary tumors of the aorta are extremely rare and the diagnosis is made most often after surgery or autopsy. Because clinical symptoms of abdominal sarcoma are similar to those of occlusive or aneurysmal disease, aortic sarcomas are frequently mistaken for these lesions. The imaging findings are frequently nonspecific and therefore do not allow a definitive preoperative diagnosis. We report a case of an epithelioid angiosarcoma in the vessel wall of an abdominal aortic aneurysm.

Aero-digestive tract squamous intra-epithelial neoplasia is a disease whose genetic and epigenetic features lead to clinical signs and well codified histologic features. This publication aims to review the molecular alterations which have been identified in these lesions, to clarify the clinical manifestations and to discuss the proposed histological classification.

Many studies have already examined the hematopoietic recovery after irradiation but paid with very little attention to the bone marrow microenvironment. Nonetheless previous studies in a murine model of reversible radio-induced bone marrow aplasia have shown a significant increase in alkaline phosphatase activity (ALP) prior to hematopoietic regeneration. This increase in ALP activity was not due to cell proliferation but could be attributed to modifications of the properties of mesenchymal stem cells (MSC). We thus undertook a study to assess the kinetics of the evolution of MSC correlated to their hematopoietic supportive capacities in mice treated with sub lethal total body irradiation. In our study, colony-forming units – fibroblasts (CFU-Fs) assay showed a significant MSC rate increase in irradiated bone marrows. CFU-Fs colonies still possessed differentiation capacities of MSC but colonies from mice sacrificed 3 days after irradiation displayed high rates of ALP activity and a transient increase in osteoblastic markers expression while pparγ and neuropilin-1 decreased. Hematopoietic supportive capacities of CFU-Fs were also modified: as compared to controls, irradiated CFU-Fs significantly increased the proliferation rate of hematopoietic precursors and accelerated the differentiation toward the granulocytic lineage. Our data provide the first evidence of the key role exerted by the balance between osteoblasts and adipocytes in spontaneous bone marrow regeneration. First, (pre)osteoblast differentiation from MSC stimulated hematopoietic precursor's proliferation and granulopoietic regeneration. Then, in a second time (pre)osteoblasts progressively disappeared in favour of adipocytic cells which down regulated the proliferation and granulocytic differentiation and then contributed to a return to pre-irradiation conditions.

Cytomegalovirus (CMV), Pneumocystis carinii , and Aspergillus species are common causes of fatal pulmonary infections in immunocompromised hosts. Therefore, rapid and reliable methods of establishing the diagnosis of these types of pneumonia are essential. Bronchoalveolar lavage (BAL) has proved to be a rapid and safe procedure for procuring large numbers of bronchial and alveolar cells from the lung. To assess various methods of detecting CMV, P. carinii , and aspergillosis in lavage specimens, 47 BAL samples from adults at high risk for these infections were evaluated. The visualization of these agents was performed by cytologic examination and in situ hybridization for CMV; cytologic examination, Gomori’s methenamine silver (GMS) stain, and immunofluorescence for P. carinii ; and cytologic examination, GMS stain, and immunocytologic studies for Aspergillus species. Cytomegalovirus was detected in 2 of 47 specimens (4%) by cytologic examination and 7 of 47 (15%) by in situ hybridization. Cells with nuclear and/or cytoplasmic inclusions invariably were labeled with the CMV DNA probe. The weak diagnostic value of the cytologic examination resultedfrom the absence of characteristic inclusions in many specimens with positive results by in situ hybridization. P. carinii was the most frequent pathogen isolated from BAL fluid (9 of 47 cases; 19%). It was found in 1 of 47 specimens (2%) by the cytologic examination of Papanicolaou- stained smears, 4 of 47 (8,5%) by the GMS stain, and 8 of 47 (17%) by immunofluorescence. Most P. carinii -positive cases (five of nine cases) were detected by immunofluorescence only. Aspergillus species was diagnosed in 2 of 47 specimens (4%) by cytologic examination and GMS staining. Immunocytologic studies had positive results in these specimens and detected one additional case of Aspergillus infection (3 of 47 cases; 6%). These data show that techniques using CMV DNA probes and anti- P. carinii or anti-Aspergillus antibodies are rapid and more sensitive than conventional diagnostic procedures.

The monoclonal antibody EB-A1 to galactomannan is apparently specific for detecting Aspergillus species and Penicillium marneffei in formalin-fixed, paraffin-embedded tissues. It reveals hyphae, remnants of filaments, and organisms in the cytoplasm of some phagocytic cells.

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.

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. In a first step, we have demonstrated that the density of ALP network and the number of ALP positive cells increase strongly in the first three days following irradiation. This increase precede haematopoietic regeneration and isn’t a result of cell proliferation. In vitro studies have also demonstrated that this increase in ALP activity is due to the expression of TNSALP (tissue non specific alkaline phosphatase). Next, we have established a relationship between these ALP positive reticular cells and mesenchymal stem cells. In vitro observations, as well as phenotypic analysis, indicate that TNSALP+ cells differentiated from mesenchymal stem cells. Although expressing an early osteoblastic marker, cbfa1, they keep self-renewal and differentiation capacities. To test their function in haematopoiesis, we have realized co-cultures of CFU-Fs and haematopoietic precursors (ckit+, sca-1+, lin-). We observed that, CFU-Fs from irradiated bone marrow, characterized by a high proportion of TNSALP positive cells, strongly stimulate the proliferation and the differentiation to the myelo-monocytic and granulopoietic lineage. In accordance with this observation, bone marrows of TNSALP deficient mice, at a late embryonic stage, are characterized by a deficit in granulopoiesis. In order to understand the effects of TNSALP in granulopoiesis, we added recombinant TNSALP (Enobia, Montreal Canada) on TNSALP negative CFU-Fs from control mice and observed that it has an effect on the differentiation process without any effects on the proliferation. All these data suggest that TNSALP expressed by reticular cells, directly derived from mesenchymal stem cells plays a role in normal haematopoiesis and in regeneration of bone marrow aplasia by inducing differentiation of myelo-monocytic and granulocytic precursors. Recombinant TNSALP (Enobia, Montreal Canada) will be tested in murine models of bone marrow aplasias.

AIM: To report the experience of the CHU Sart Tilman, University of Liege, Belgium, in the management of appendiceal carcinoid tumor. METHODS: A retrospective review of 1237 appendectomies performed in one single centre from January 2000 to May 2004, was undertaken. Analysis of demographic data, clinical presentation, histopathology, operative reports and outcome was presented. RESULTS: Among the 1237 appendectomies, 5 appendiceal carcinoid tumors were identified (0.4%) in 4 male and 1 female patients, with a mean age of 29.2 years (range: 6-82 years). Acute appendicitis was the clinical presentation for all patients. Four patients underwent open appendectomy and one a laparoscopic procedure. One patient was reoperated to complete the excision of mesoappendix. All tumors were located at the tip of the appendix with a mean diameter of 0.6 cm (range: 0.3-1.0 cm). No adjuvant therapy was performed. All patients were alive and disease-free during a mean follow-up of 33 mo. CONCLUSION: Appendiceal carcinoid tumor most often presents as appendicitis. In most cases, it is found incidentally during appendectomies and its diagnosis is rarely suspected before histological examination. Appendiceal carcinoid tumor can be managed by simple appendectomy and resection of the mesoappendix, if its size is ≤ 1 cm.

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.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.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.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.

Purpose: To assess, in lung cancer patients, the effects of thoracic radiotherapy (RT) on the concentrations of transforming growth factor-β1 (TGF-β1) and interleukin-6 (IL-6) in the bronchoalveolar lavage (BAL) fluid. Methods and Materials: Eleven patients with lung cancer requiring RT as part of their treatment were studied. BAL was performed bilaterally before, during, and 1, 3, and 6 months after RT. Before each BAL session, the patient's status was assessed clinically using pulmonary function tests and an adapted late effects on normal tissue-subjective, objective, management, analytic (LENT-SOMA) scale, including subjective and objective alterations. The National Cancer Institute Common Toxicity Criteria were used to grade pneumonitis. The TGF- β1 and IL-6 levels in the BAL fluid were determined using the Easia kit. Results: The TGF-β1 and IL-6 concentrations in the BAL fluid recovered from the irradiated areas were significantly increased by thoracic RT. The increase in TGF-β1 levels tended to be greater in the group of patients who developed severe pneumonitis. In the BAL fluid from the nonirradiated areas, the TGF-β1 and IL-6 concentrations remained unchanged. Conclusion: The observed increase in TGF-β1 and IL-6 concentrations in the BAL fluid recovered from the irradiated lung areas demonstrated that these cytokines may contribute to the process leading to a radiation response in human lung tissue.

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.

Isolated gonadotropin deficiency due to inactivating mutations of -subunits of LH or FSH are rare. Isolated LH inactivation was identified in1992,although thepatient’s clinical characteristicshad been reportedearlier (1,2).Maleswith inactivatingLH mutations (five adults have been reported) present with clinical features of hypogonadism and oligo/azoospermia (1–5). In 2004, we reported the case of a 30-yr-old man with this clinical presentation (3). The patient gave informed consent for a testicular biopsy (Fig. 1, A and B), which showed arrested spermatogenesis and fetal-type Leydig cells (3). A homozygous missense mutation (G36D) in the LH -subunit gene was identified that abrogated -subunit dimerization and rendered LH biologically and immunologically inactive (3). We initiated treatment with intramuscular human chorionic gonadotropin (hCG) (1500 IU three times a week for 1 month, then 5000 IU weekly). Tanner staging was 5 after 3 months. After 24 months treatment, FSH decreased to 2.3 mIU/ml (normal range, 1–8mIU/ml)andtestosterone increasedto7 g/liter (normal range, 2.5–10.0 g/liter).Therewasnearnormalizationof testicular structure, which was likely related to Leydig cell maturation and subsequent increases in intratesticular testosterone, leading to a sperm count of 1000 spermatozoids/ml. (Fig. 1, C and D). The patient and his wife conceived a child by intracytoplasmic sperm injection from ejaculated sperm. The male child was heterozygotic for the G36D LH mutation and was phenotypically normal with normal LH, FSH, and testosterone levels at the age of 4 wk. Although rare, isolated LH deficiency due to inactivating mutations of LH -subunit gene is a useful illustration of the precise role of LH in testicular maturation and function in humans. Furthermore, it also provides a good example of the clinical efficacy of LH receptor stimulation using hCG.

A 37-year-old man with acute myeloblastic leukemia in first remission developed ulcerative colitis and bronchiolitis obliterans organizing pneumonia (BOOP) 7 months after bone marrow transplantation (BMT) from an HLA-matched brother who suffered from severe Crohn's disease. BOOP occurred 20 days after idiopathic interstitial pneumonia, in the context of severe ulcerative colitis. Lung and colon biopsies showed no signs of CMV infection or GVHD. The patient was treated with oral methylprednisolone 1 mg/kg/day and his clinical status and chest X-ray improved slowly. Remarkably, the symptoms of colitis also resolved with prednisone therapy and he is now symptom-free. We hypothesize that ulcerative colitis may have been transmitted from donor to recipient (adoptive autoimmunity) and that it was complicated by BOOP. However, other factors such as CMV may have contributed to the occurrence of BOOP.

Bone marrow specimens from 118 patients with clinical suspicion of multiple myeloma were studied to assess the diagnostic reliability of histological and cytological criteria. Plasma cell clonality was assessed by demonstrating light chain restriction. In most cases of multiple myeloma, the classical cytological and histological criteria were found. In eight cases in which the marrow contained less than 10% of plasma cells, there was discordance between the light microscopic and immunohistochemical findings. In six cases without evidence of multiple myeloma and with polyclonal plasma cells in bone marrow, abnormal plasma cells resembling a malignant proliferation were found. These findings indicate that comparison of histological and cytological results with immunohistological studies for immunoglobulin light chains in bone marrow biopsy sections can be helpful in the evaluation of patients with a suspicion of multiple myeloma and when the marrow contains less than 10% of plasma cells.

Fractionated whole-body irradiation induces thymic lymphomas in most of treated C57Bl/Ka mice. The cellular events occurring during the latency period consist of the emergence of preleukaemic cells and of marked alterations to the T-cell lineage and the microenvironment within the thymus. The proportions of the various thymocyte subsets are modified, suggesting a blockage in the normal differentiation process. Thymic epithelial cells are functionally modified, leading to decreased interactions with immature thymocytes. Interestingly, bone marrow grafting early after irradiation, which inhibits the development of lymphomas, induces the disappearance of preleukaemic cells from the thymus, whereas thymocyte subpopulations and thymic epithelium are restored. Interferon gamma and tumor necrosis factor alpha also prevent the onset of lymphomas. Studies on the effect of bone marrow transplantation and cytokine inoculation in split-dose irradiated mice should allow characterization of the factors that modulate the progression of preleukaemic cells towards the neoplastic state.