glycogen and Sarcoma--Ewing

Ewings' sarcoma involving the facial bones is rare although it is the second most frequent bone malignancy. For the diagnosis a biopsy is needed. Currently the treatment is the combination of chemotherapy and radiotherapy with surgery indicated in only a few instances. We report a case of Ewing's sarcoma of the mandible and describe its clinicopathologic features and the treatment of this disease.

Topics: 12E7 Antigen; Adolescent; Antigens, CD; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Cell Adhesion Molecules; Combined Modality Therapy; Female; Glycogen; Humans; Mandibular Neoplasms; Sarcoma, Ewing; Vimentin

A 20-year-old woman presented with nasal obstruction and slight epistaxis. The obstructing lesion was excised and microscopy showed a neoplasm composed of comparatively uniform undifferentiated cells forming solid nests. The cytoplasm of the cells was clear but poorly demarcated, partly vacuolated and contained much glycogen. Although widespread in the nasal mucosa, the cells did not penetrate into the underlying bone. The cells expressed the MIC2 gene (using the CD99 marker). Electron microscopy showed simple cells with a small number of mitochondria, many glycogen particles; there were no neurosecretory granules present. Early surgical treatment followed by chemo- and radiotherapy have greatly improved the prognosis of EWS: extraskeletal Ewing's sarcoma (EWS/PNET).

Topics: 12E7 Antigen; Adult; Antigens, CD; Cell Adhesion Molecules; Female; Glycogen; Humans; Immunohistochemistry; Microscopy, Electron; Nasal Mucosa; Nose Neoplasms; Sarcoma, Ewing; Vimentin

Topics: Adolescent; Adult; Age Factors; Bone Neoplasms; Child; Child, Preschool; Diagnosis, Differential; False Positive Reactions; Glycogen; Histocytochemistry; Humans; Prognosis; Sarcoma, Ewing; Staining and Labeling

Glycogen dysregulation is a hallmark of aging, and aberrant glycogen drives metabolic reprogramming and pathogenesis in multiple diseases. However, glycogen heterogeneity in healthy and diseased tissues remains largely unknown. Herein, we describe a method to define spatial glycogen architecture in mouse and human tissues using matrix-assisted laser desorption/ionization mass spectrometry imaging. This assay provides robust and sensitive spatial glycogen quantification and architecture characterization in the brain, liver, kidney, testis, lung, bladder, and even the bone. Armed with this tool, we interrogated glycogen spatial distribution and architecture in different types of human cancers. We demonstrate that glycogen stores and architecture are heterogeneous among diseases. Additionally, we observe unique hyperphosphorylated glycogen accumulation in Ewing sarcoma, a pediatric bone cancer. Using preclinical models, we correct glycogen hyperphosphorylation in Ewing sarcoma through genetic and pharmacological interventions that ablate in vivo tumor growth, demonstrating the clinical therapeutic potential of targeting glycogen in Ewing sarcoma.

Topics: Animals; Bone Neoplasms; Child; Glycogen; Humans; Male; Mice; Osteosarcoma; Sarcoma, Ewing; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Extra-osseous Ewing sarcomas/peripheral primitive neuroectodermal tumors (EOES/pPNETs) are high-grade malignant tumors found in various organs, such as the lung, skin, intestine, kidney and female genital tract; however, to the best of our knowledge, only two cases have previously been identified in the thyroid gland. We describe a case of primary EOES/PNET of the thyroid gland in a 66-year-old man with a previous history of large B cell lymphoma. During a routine follow-up examination, the patient underwent an ultrasound cervical scan showing a solid nodule of the left thyroid lobe. The fine-needle aspiration biopsy of the nodule suggested a neuroendocrine tumor. Histological and immunohistochemical examination of the surgical specimen supported a diagnosis of EOES/PNET, which was further confirmed by the demonstration of EWSR1 gene translocation by means of fluorescent in situ hybridization and by the detection of glycogen particles and neurosecretory granules by means of electron microscopy. Total body computed tomography and magnetic resonance imaging excluded the involvement of other sites, and therefore a diagnosis of primary EOES/PNET of the thyroid gland was made.This paper also discusses the main differential diagnoses, including lymphoma recurrence, other small round cell tumors (primary or metastatic), and a thyroid localization of an EWS/PNET from another organ.

Topics: Aged; Biomarkers, Tumor; Biopsy, Needle; Calmodulin-Binding Proteins; Diagnosis, Differential; DNA, Neoplasm; Glycogen; Humans; In Situ Hybridization, Fluorescence; Incidental Findings; Lymph Nodes; Lymphoma, Large B-Cell, Diffuse; Male; Neoplasm Recurrence, Local; Neuroectodermal Tumors, Primitive, Peripheral; RNA-Binding Protein EWS; RNA-Binding Proteins; Sarcoma, Ewing; Secretory Vesicles; Thyroid Neoplasms; Translocation, Genetic

Ewing family of tumors (EFTs) comprise highly malignant, nearly undifferentiated neoplasms including Ewing's sarcoma (ES), primitive neuroectodermal tumor (PNET) and a spectrum of other unusual variants. In general, EFTs are included among small blue cell tumors. Establishing histological diagnosis can be difficult; CD99 and FLI1 immunohistochemical staining has improved diagnosis, but these markers are not specific for EFTs. The diagnosis of EFTs is confirmed by molecular diagnostic testing showing the presence of established rearrangements of the EWS gene. The use of this molecular signature in EFTs revealed rare variants in the histomorphologic spectrum of these tumors. The authors report an unusual variant of EFT in the mandible of a 17-year-old patient, which was confirmed by translocation rearrangement in EWR1 gene at 22q12 by fluorescence in situ hybridization. The unusual histologic features, with prominent spindling of tumor cells and deviation from the classic features of Ewing's sarcoma posed a diagnostic challenge for the medical centers involved in the diagnosis and treatment of this patient. This highlights the importance of the genetic study of undifferentiated sarcomas to identify rare morphologic variants of ES, in view of the chemosensitivity of EFTs and how this affects patient management.

Topics: 12E7 Antigen; Adolescent; Antigens, CD; Carboxylesterase; Cell Adhesion Molecules; Chondrosarcoma, Mesenchymal; Chromosomes, Human, Pair 22; Diagnosis, Differential; Female; Follow-Up Studies; Gene Rearrangement; Glycogen; Humans; In Situ Hybridization, Fluorescence; Mandibular Neoplasms; RNA-Binding Protein EWS; Sarcoma; Sarcoma, Ewing; Translocation, Genetic

We report a case of Ewing sarcoma/peripheral primitive neuroectodermal tumor (ES/pPNET) arising in the vulva in a 17-year-old adolescent girl. Primary extraskeletal ES/pPNET is uncommon and only rarely affects the female genital tract. Characteristic histologic features of ES/pPNET were present in this case including a monomorphic population of small round blue cells with cytoplasmic glycogen confirmed by periodic acid-Schiff and were immunohistochemically positive for CD99 in a membranous and Fli-1 in a nuclear pattern. The presence of an EWS/Fli-1 fusion transcript was demonstrated by reverse transcription polymerase chain reaction and confirmed by direct sequencing. This report described another very rare case of ES/pPNET arising in the vulva and confirmed by molecular analysis.

Topics: Adolescent; Base Sequence; Biomarkers, Tumor; DNA, Neoplasm; Female; Glycogen; Humans; Molecular Sequence Data; Neoplasms, Multiple Primary; Neuroectodermal Tumors, Primitive, Peripheral; Oncogene Proteins, Fusion; Periodic Acid-Schiff Reaction; Proto-Oncogene Protein c-fli-1; RNA-Binding Protein EWS; Sarcoma, Ewing; Sequence Analysis, DNA; Transcription Factors; Vulvar Neoplasms

Extraskeletal Ewing's sarcoma is a rare tumor. The most common sites of occurrence are on the trunk, extremities, and retroperitoneum. This type of tumor is well characterized by recurrent chromosomal translocation such as t (11;22) (q24;q12) (EWSR1/FLI1) or t (21;22) (q22;q12) (EWSR1/ERG) and overexpression of MIC2/CD99 on tumor cell membrane. We describe the first reported case of an esophageal extraskeletal Ewing's sarcoma with confirmation from immunohistochemical and molecular diagnoses. A 56-year-old man developed a polypoid tumor located in the lower part of the esophagus. The tumor was composed of small-sized round cells showing prominent fibrillar cytoplasmic processes. Intracytoplasmic glycogen was detected in all the tumor cells. Immunoreactivity for MIC2/CD99 was positive on the membrane of all tumor cells. A reverse transcriptase-polymerase chain reaction followed by sequencing revealed an EWSR1/ERG chimeric transcript, which combined EWSR1 exon 10 with ERG exon 6. The present report added a new entity of esophageal small round cell tumor.

Topics: 12E7 Antigen; Antigens, CD; Bone Neoplasms; Cell Adhesion Molecules; DNA, Neoplasm; Esophageal Neoplasms; Glycogen; Humans; Immunoenzyme Techniques; Male; Middle Aged; Oncogene Proteins, Fusion; Proto-Oncogene Protein c-fli-1; Reverse Transcriptase Polymerase Chain Reaction; RNA-Binding Protein EWS; Sarcoma, Ewing; Transcription Factors; Translocation, Genetic

Ewing's sarcoma shows a strong tendency to metastasize to the lungs or the skeleton, or both. A peculiar feature of the secondary involvement of bone with this tumor is that it may also appear in the absence of clinically evident lung metastases, both at clinical presentation and during the course of the disease. Although osseous metastases are critically relevant for prognosis, the pathogenesis of this peculiar feature of Ewing's sarcoma is poorly understood, partly due to the lack of appropriate experimental in vivo models. We show that the intravenous injection of TC-71 Ewing's sarcoma cells into athymic 4-5-week-old Crl/nu/nu (CD1) BR mice reproducibly colonizes specific sites of the skeleton in addition to the lungs and lymph nodes. The distribution and the morphologic appearance of these experimental bone metastases mimic the pattern of skeletal involvement observed in humans. This experimental model of bone metastasis of Ewing's sarcoma may be the basis for future studies aimed at understanding the pathophysiology and treatment of Ewing's sarcoma.

Topics: 12E7 Antigen; Animals; Antigens, CD; Bone Neoplasms; Cell Adhesion Molecules; Disease Models, Animal; Female; Glycogen; Integrins; Lung Neoplasms; Matrix Metalloproteinases; Mice; Mice, Nude; Neoplasm Metastasis; Receptors, Growth Factor; Sarcoma, Ewing; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinase-2; Tumor Cells, Cultured

Topics: 12E7 Antigen; Actins; Antigens, CD; Bone Neoplasms; CD3 Complex; Cell Adhesion Molecules; Child; Diagnosis, Differential; Diagnostic Errors; Glycogen; Humans; Immunohistochemistry; Intercellular Junctions; Leukocyte Common Antigens; Liability, Legal; Lymphoma; Microscopy, Electron; Neuroectodermal Tumors, Primitive; Pathology; Phosphopyruvate Hydratase; Sarcoma, Ewing; Vimentin

The differential diagnosis of cutaneous small round cell malignancies is a relatively uncommon but recurrent problem that usually requires adjuvant techniques including special histochemical stains, immunohistochemistry (IHC), electron microscopy (EM), and cytogenetics (CG) to arrive at a definite answer. This report describes a case of a primary cutaneous malignancy that, after workup, fulfilled the criteria of extraskeletal Ewing's family sarcoma, which was corroborated by IHC with an antibody to glycoprotein p30/32 mic2 that is highly expressed in these neoplasms. The lesions consisted of a large nodular proliferation of poorly differentiated monotonous small round cells confined to the dermis and subcutaneous tissue. The cells had high nuclear to cytoplasmic (N/C) ratios, scattered prominent nucleoli, and indistinct cytoplasm. A periodic acid-Schiff (PAS) stain with and without diastase demonstrated abundant cytoplasmic glycogen. The glycogen was confirmed with EM, which did not show neurosecretory granules, but extensive sectioning of the tissue blocks demonstrated with light microscopy a single focus with pseudorosette formation. IHC was positive for monoclonal antibody (MAb) O13 to glycoprotein p30/32 mic2 and negative for lymphoid (CD45), neural (S-100, NF, GFAP), neuroendocrine (NSE), and muscle (MSA, desmin) markers. To the best of our knowledge, this is one of few reported cases of primary cutaneous (extraskeletal/extraosseous) Ewing's sarcoma (EEWS) and the first to use IHC with MAb O13, which recognizes the cell surface glycoprotein p30/32 mic2. This case further illustrates the continuum between EEWS and primitive peripheral neuroepithelioma and supports the unifying concept that these two entities are merely subtle morphologic variants of the same malignant neoplasm, which is better designated a Ewing's family sarcoma.

Topics: 12E7 Antigen; Actins; Adolescent; Antibodies, Monoclonal; Antigens, CD; Biomarkers, Tumor; Cell Adhesion Molecules; Cell Differentiation; Cell Division; Cell Nucleolus; Cell Nucleus; Coloring Agents; Cytogenetics; Cytoplasm; Desmin; Diagnosis, Differential; Female; Glial Fibrillary Acidic Protein; Glycogen; Humans; Immunohistochemistry; Leukocyte Common Antigens; Microscopy, Electron; Neurofilament Proteins; Phosphopyruvate Hydratase; S100 Proteins; Sarcoma, Ewing; Skin; Skin Neoplasms

Topics: Adolescent; Antibodies, Monoclonal; Antigens, Neoplasm; Biomarkers, Tumor; Bone Neoplasms; Child; Glycogen; Humans; Sarcoma, Ewing

Recent reports of Ewing's sarcoma (EW) and extraskeletal Ewing's sarcoma (EEW) support the hypothesis that these tumors are neuroectodermal in origin. Primitive neuroectodermal tumors (PNET) of bone (32 cases) and soft tissue (25 cases) including those previously categorized as EW in 27 cases and EEW in 15 cases were carefully studied histologically, immunocytochemically and morphometrically, focusing on tumor cell differentiation. This study attempts to subclassify these tumors on the basis of the size of tumor cells and nuclei, their variations (uniformity or diversity), arrangement of tumor cells (rosette or non-rosette), focal differentiation to larger ganglion-like cells, and staining intensity for neural markers. All tumors were histologically subclassified as small, medium or large cell types, three basic subtypes (rosette type, abortive rosette type, non-rosette type) and four complementary subtypes (fibrillary type, non-fibrillary type, angiomatoid type, ganglion cell type). Classic EW or EEW is consistent with small or medium, non-rosette, non-fibrillary type tumors, previously described large cell EW with large, non-rosette, fibrillary or non-fibrillary type tumors, and classic neuroectodermal tumor with small or medium, rosette, fibrillary type tumors, according to the present subclassification. Clinicopathologic correlations with the different subtypes are discussed. Long-term survival, more than 5 years, was seen in patients with small cell type, and those younger than 14 years of age.

Topics: Adolescent; Adult; Bone Neoplasms; Child; Child, Preschool; Female; Follow-Up Studies; Glycogen; Humans; Infant; Male; Neoplasms, Germ Cell and Embryonal; Neurofilament Proteins; Phosphopyruvate Hydratase; Sarcoma, Ewing; Soft Tissue Neoplasms; Survival Rate

The authors report the case of an 8-year-old boy with a Ewing's sarcoma localized to the mandible. The therapeutic modalities consisting of induction chemotherapy, surgical removal of the involved portion of the mandible with reconstruction using a parascapular osteo-cutaneous free flap, and maintenance chemotherapy are described. Emphasis is placed on the importance of performing a wide resection and the use of a multidisciplinary team approach in the treatment of these unusual tumours.

Topics: Cell Nucleolus; Cell Nucleus; Child; Chromatin; Cytoplasm; Glycogen; Humans; Male; Mandibular Neoplasms; Sarcoma, Ewing; Vimentin

This study describes functional characteristics of receptors for vasoactive intestinal peptide (VIP) on human Ewing's sarcoma WE-68 cells. These characteristics include 125I-VIP binding capacity, cellular cAMP generation, glycogen hydrolysis, and pharmacological specificity. Binding studies with 125I-VIP showed specific, saturable, binding sites for VIP in WE-68 cells. Scatchard analysis revealed the presence of a single class of high-affinity binding sites that exhibited a dissociation constant (Kd) of 90 pM and a maximal binding capacity (Bmax) of 24 fmol/mg of protein. VIP and VIP-related peptides competed for 125I-VIP binding in the following order of potency: human (h) VIP greater than human peptide with N-terminal histidine and C-terminal methionine (PHM) greater than chicken secretin much greater than porcine secretin. Glucagon and the C-terminal fragments VIP[10-28] and VIP[16-28] and the VIP analogue (D-Phe2)VIP did not inhibit 125I-VIP binding. Addition of hVIP to WE-68 cells provoked marked stimulation of cAMP accumulation, hVIP stimulated increases in cAMP content were rapid, concentration-dependent, and potentiated by 3-isobutyl-l-methylxanthine (IBMX). Half-maximal stimulation (EC50) occurred at 150 nM hVIP. The ability of hVIP and analogues to stimulate cAMP generation paralleled their potencies in displacing 125I-VIP binding. (D-Phe2)VIP, VIP[10-28], VIP[16-28], and (p-Cl-D-Phe6, Leu17)VIP, a putative VIP receptor antagonist, affected neither basal cAMP levels nor hVIP-induced cAMP accumulation. WE-68 cell responses to hVIP were desensitized by prior exposure to hVIP. Desensitization to hVIP did not modify the cAMP response to beta-adrenergic stimulation, and beta-adrenergic agonist desensitization did not modify responses to hVIP. hVIP also induced a time- and concentration-dependent hydrolysis of 3H-glycogen newly formed from 3H-glucose in WE-68 cultures. hVIP maximally decreased 3H-glycogen content by 36% with an EC50 value of about 8 nM. The order of potency of structurally related peptides of hVIP for stimulation of glycogenolysis correlated with their order of potency for inhibition of 125I-VIP binding. IBMX potentiated the glycogenolytic action of hVIP and PHM. The simultaneous presence of the calcium channel antagonist verapamil or the calcium ionophore A 23187 did not influence the glycogenolytic and cAMP stimulatory effects of hVIP. Collectively, these data indicate that Ewing's sarcoma (WE-68) cells are endowed with genuine VIP

Topics: Cyclic AMP; Glycogen; Humans; Iodine Radioisotopes; Peptide Fragments; Protein Binding; Receptors, Gastrointestinal Hormone; Receptors, Vasoactive Intestinal Peptide; Sarcoma, Ewing; Tumor Cells, Cultured; Vasoactive Intestinal Peptide

This study describes hormonal regulation of glycogen metabolism in Ewing's sarcoma cells. 3H-Glycogen synthesized in cultured Ewing's sarcoma WE-68 cells from 3H-glucose was hydrolyzed in a concentration-dependent manner by various catecholamines. The order of potency for the glycogenolytic effects of catecholamines was isoproterenol greater than or equal to dopamine greater than norepinephrine greater than epinephrine. The concentrations giving half-maximal effectiveness (EC50) were about 2 x 10(-8) M, 3 x 10(-8) M, 8 x 10(-8) M, and 5 x 10(-7) M for isoproterenol, dopamine, norepinephrine, and epinephrine, respectively. Higher concentrations of each of the catecholamines were necessary to elicit EC50 stimulation of cyclic AMP production in Ewing's sarcoma cells. Glycogenolysis induced by dopamine was blocked by chlorpromazine, a dopamine D1-receptor antagonist, but not by haloperidol, a dopamine D2-receptor antagonist. The glycogenolytic action of norepinephrine was markedly reduced by propranolol, a beta-adrenoreceptor antagonist, and was not affected by yohimbine, an alpha-adrenoreceptor antagonist. In addition, chlorpromazine also antagonized the glycogenolytic response to norepinephrine. Dibutyryl cyclic AMP, 3-isobutyl-1-methylxanthine, and the diterpene forskolin were also found to induce 3H-glycogen hydrolysis. Our data indicate that catecholamines exert their glycogenolytic effects in Ewing's sarcoma cells by stimulation of cyclic AMP formation via beta-adrenergic receptors and dopamine D1-receptors.

Topics: Adrenergic beta-Agonists; Animals; Cell Line; Cells, Cultured; Chlorpromazine; Cyclic AMP; Dopamine; Dose-Response Relationship, Drug; Glycogen; Haloperidol; Norepinephrine; Sarcoma, Ewing; Time Factors

Ewing's sarcoma (ES) of bone may occasionally display rosette-like textures mimicking Homer-Wright ones, as seen in neuroectodermic neoplasms (neuroblastoma, peripheral neuroepithelioma). Of a group of 39 cases of ES, reviewed with electron microscopic study, the authors have isolated five atypical ES, which histologically also possessed neuroectodermic traces. These tumors were composed of small round blue cells with rosette-like figures and cytoplasmic glycogen. The immunohistochemical analysis showed positivity for neuron-specific enolase (NSE) as well as for HNK-1 (leu-7) monoclonal antibody. Electron microscopic examination confirmed the tumor cell as being of small round type, with a dense chromatine pattern and the presence of isolated dendritic processes, as well as synaptic-like buttons; intermediate filaments, neurotubuli, and dense-core neurosecretory granules also were seen. Moreover, in two cases basement-like condensations surrounded some cells. Scanning electron microscopic study in one case confirmed the presence of rosette-like figures and cell elongations with short dendritic projections of the cytoplasm. Clinically and radiologically these cases showed features similar to ES of bone; one case, located in the chest wall, had a local relapse after treatment, with the histologic features of a pleomorphic neuroblastoma. The authors conclude that these tumors resemble closely immature neuroepithelioma of soft tissue but, being primary to bone, are superimposable on those described as "neuroectodermal tumors of bone."

Topics: Antibodies, Monoclonal; Antigens, Differentiation, T-Lymphocyte; Antigens, Surface; Bone Neoplasms; Glycogen; Histocytochemistry; Immunologic Techniques; Microscopy, Electron; Microscopy, Electron, Scanning; Neuroblastoma; Neuroectodermal Tumors, Primitive, Peripheral; Peripheral Nervous System Neoplasms; Phosphopyruvate Hydratase; Sarcoma, Ewing

Topics: Adult; Glycogen; Humans; Immunohistochemistry; Male; Sarcoma, Ewing; Soft Tissue Neoplasms; Thoracic Neoplasms; Vimentin

In 1969 it was recognised that tumors with light microscopic appearances indistinguishable from Ewing's sarcoma of bone may arise in extraskeletal sites (extraskeletal Ewing's sarcoma). Here, we review the available literature and report five new cases. All five received combined modality therapy with combination chemotherapy and radiotherapy to the primary site followed by surgical excision in two. All attained complete remission; after a median follow-up of 26 months, three remain disease-free but two have relapsed and died. Our experience, in accord with previous series, suggests that extraskeletal Ewing's sarcoma compared with its bony counterpart tends to occur in older subjects, has a similar incidence in males and females, usually presents with a painless mass and readily responds to combined modality therapy. We detected no light or electron microscopic features to denote a histogenetic origin. However, we suspect extraskeletal Ewing's sarcoma may occur more frequently than previously supposed.

Topics: Adult; Antineoplastic Combined Chemotherapy Protocols; Combined Modality Therapy; Female; Follow-Up Studies; Glycogen; Humans; Male; Microscopy, Electron; Sarcoma, Ewing

Ewing's sarcoma is a rare, but highly malignant bone tumor. During the years 1966-1983 we diagnosed and treated this tumor in 84 patients. Diagnosis of the tumor in an early stage is a prerequisite for successful therapy. Clinical and radiological findings contribute to the diagnosis, but histologic confirmation is indispensible. In addition to histomorphology we regard electron microscopy and the demonstration of glycogen as essential for the final diagnosis. The differential diagnosis includes haematogeneous osteomyelitis, malignant lymphoma, metastases from neuroblastomas, rhabdomyosarcoma and some benign bone tumors. New observations concerning the histogenesis of Ewing's sarcoma are discussed. The introduction of chemotherapy as part of a multimodality approach to treatment has substantially improved the prognosis in comparison to the previous results with surgery and radiation therapy. The role of surgery and radiation independently and in combination is discussed. We propose an approach to treatment based on our experience which involves surgical removal of the primary tumor after preoperative chemotherapy. The overall survival rate among our 84 patients was 40%. Among those who were diagnosed early and treated according to the recommended protocol 54% are alive.

Topics: Adolescent; Adult; Antineoplastic Combined Chemotherapy Protocols; Child; Child, Preschool; Combined Modality Therapy; Diagnosis, Differential; Female; Glycogen; Humans; Infant; Male; Microscopy, Electron; Middle Aged; Prognosis; Sarcoma, Ewing

Four small round cell malignant tumors of bone occurring in children are described. There was no catecholamine secretion and the clinical, radiologic, and biopsy diagnosis in each was Ewing's sarcoma. Glycogen was sparse both on imprints and in tissue sections. The tumors, when extensively sampled, had areas of a lobular growth pattern and Homer Wright rosettes. The rosettes were always focal and varied in complexity from case to case; they were rudimentary in one instance and markedly fibrillar in the most obvious instance. Neuron-specific enolase was demonstrated in tissue sections and in longterm cell cultures from three of the tumors. The cultured cells put out moderately long beaded processes in serum-free medium but had no catecholamine fluorescence. Electron microscopy of the tumor rosettes and the cultured cells showed processes containing aggregates of microtubules and only one case had rare neurosecretory granules. This study suggests that some small round cell tumors of bone and soft tissue in children, which present as Ewing's sarcoma, are neuroectodermal in nature.

Topics: Bone Neoplasms; Calcaneus; Child; Child, Preschool; Female; Femoral Neoplasms; Glycogen; Humans; Male; Microscopy, Electron; Neuroblastoma; Neuroectodermal Tumors, Primitive, Peripheral; Phosphopyruvate Hydratase; Rosette Formation; Sarcoma, Ewing; Scapula; Staining and Labeling

Forty-two cases of Ewing's sarcoma (ES) have been studied with light microscopy during the 9-year period 1974 to 1982. Thirty-three patients had ES of bone, and in 9 patients the tumor was located in the extraskeletal soft tissues. Cases which fulfilled all the morphologic criteria were accepted as typical ES (31 cases), and those with some architectural or cytologic peculiarities were considered atypical forms of ES (11 cases). An immunohistochemical study (PAP method) to evaluate the presence in the tumor cells of the following markers: myoglobin, F-VIII-related antigen, lysozyme, alpha-1-antitrypsin, alpha-1-antichymotrypsin, and immunoglobulins (IgG, IgA, IgM, kappa and lambda light chains), was performed with negative results in all cases (paraffin blocks were available in 38 cases). The cytochemical study on fresh tissue imprints from five patients (PAS, Sudan Black, alpha-naphthyl acetate esterase, acid phosphatase, beta glucuronidase, myeloperoxidases, naphthol-AS-D chloroacetate esterase and alkaline phosphatase) gave no pattern of histogenetic significance, PAS being the best morphologic marker in tissue sections and touch preparations. A detailed ultrastructural study was performed on 34 cases; the main findings may be summarized as follows: medium sized cells, polygonal shape, oval nuclei, smooth nuclear envelope, abundant euchromatin, well-developed nucleolonema, scant membranous organelles, abundant hyaloplasmic glycogen, occasional lipid vacuoles, straight cell membranes, and primitive intercellular junctions. No differences were found between bone and extraskeletal ES; moreover, typical and atypical forms showed moderate quantitative differences with no qualitative change. The histogenesis is discussed; no functional or morphologic markers have been found to suggest the cell of origin; however, some cell lines may be excluded. It is the impression of the authors that they are dealing with a primitive noncommitted mesenchymal cell.

Topics: Adolescent; Adult; Bone Neoplasms; Cell Membrane; Cell Nucleolus; Cell Nucleus; Child; Child, Preschool; Female; Glycogen; Histocytochemistry; Humans; Immunoenzyme Techniques; Male; Microscopy, Electron; Sarcoma, Ewing; Soft Tissue Neoplasms

A case of extraskeletal Ewing's sarcoma was studied ultrastructurally, cytologically, and cytochemically. This tumor was, as in that of osseous origin, composed of densely packed highly undifferentiated blastemic cells and so-called "darker cells." The principal tumor cells were undifferentiated cells, which were characterized by the scant cytoplasm containing a variable amount of glycogen granules, large round or oval nuclei with dispersed chromatin, and rudimentary cytoplasmic junctional complexes. Some of the darker cells seemed to show somewhat histiocytic differentiation from principal blastemic cells, but the others were degenerating. Cytochemically, no tumor cells revealed the presence of peroxidase, lysozymes, and Factor VIII-related antigen. It is likely that both Ewing's sarcomas of osseous and extraosseous origin are defined as undifferentiated sarcoma merely characterized by the presence of glycogen.

Topics: Adolescent; Antigens; Cytoplasmic Granules; Diagnosis, Differential; Factor VIII; Female; Glycogen; Humans; Immunoenzyme Techniques; Microscopy, Electron; Muscles; Sarcoma, Ewing; Soft Tissue Neoplasms; von Willebrand Factor

Topics: Adult; Diagnosis, Differential; Epidural Space; Female; Glycogen; Humans; Male; Middle Aged; Sarcoma, Ewing; Spinal Canal; Spinal Cord Compression

The history of a 6-year-old girl with a tumor originating from thoracic spine and finally becoming resistant to surgery, radio-, and chemotherapy is reported. Tumor-biopsy material was studied by light and electron microscopy, in cell culture, by acetylcholinesterase ultracytochemistry, and by quantitative catecholamine analysis and this led to the rejection of the initial diagnosis of a neuroblastoma. Light microscopy revealed a uniform population of undifferentiated cells incompletely lobulated by broad fibrovascular septa. Using the electron microscope, cells were characterized by large intracellular pools of glycogen, little cytoplasm with an abundance of free ribosomes and a paucity of organelles. A few cells displayed desmosome-like attachment sites. Staining for specific and unspecific acetylcholinesterase was negative with light and electron microscopy, as were the results of catecholamine histofluorescence using the glyoxylic acid method. The latter result was confirmed by the negative outcome of quantitative analyses of dopamine, noradrenaline, and adrenaline with high pressure liquid chromatography nd electrochemical detection in tissue samples. Tumor cells could easily be maintained in culture for up to 4 weeks. None of a variety of treatments that are known to favor expression of neuronal characteristics in neuroblastoma cells (serum withdrawal, nerve growth factor, dbcAMP, dexamethasone) induced morphological differentiation in cultured tumor cells. On the basis of the clinical history, morphology, and of our experiments with tumor cells, the diagnosis of a so-called extraskeletal Ewing's sarcoma is most likely. Our results strengthen the view that a cell biology approach may be valuable in neuroblastoma differential diagnosis.

Topics: Acetylcholinesterase; Catecholamines; Cells, Cultured; Child, Preschool; Diagnosis, Differential; Female; Glycogen; Humans; Neuroblastoma; Sarcoma, Ewing

A light- and electron-microscopic review of 40 cases diagnosed at Children's Hospital of Pittsburgh as ganglioneuroblastoma, neuroblastoma, or small round cell tumor-probably neuroblastoma disclosed four cases that contained abundant glycogen. Two were unquestionable neuroblastomas by electron microscopy; one was primary in the adrenal gland, the other in the mediastinum. In the third case, a paraspinal tumor, the light-microscopic appearance was suggestive or neuroblastoma, but no catecholamine granules or neural processes were demonstrated in the material available for electron microscopy. The fourth case was an undifferentiated malignant tumor in the pectoralis muscle of a 12-year-old girl. By electron microscopy, neural processes were demonstrated and the tumor was classified as peripheral neuroblastoma. Of the remaining 36 cases, electron microscopy readily indicated a diagnosis of neuroblastoma or glangioneuroblastoma in 35 of them. In the other case, the tissue had been fixed in formalin and only a few catecholamine granules were found after an extensive search.

Topics: Adolescent; Adrenal Gland Neoplasms; Child; Child, Preschool; Diagnosis, Differential; Female; Ganglioneuroma; Glycogen; Humans; Infant; Infant, Newborn; Male; Neuroblastoma; Sarcoma, Ewing

The characteristics of 35 round cell sarcomas of bones are analyzed by optical and electron microscopical means, 28 cases of Ewing's sarcoma and 7 cases of malignant lymphomas (reticulum cell sarcoma) being distinguished. The existence of a morphological diversity within the Ewing's sarcoma group allows the authors to differentiate a conventional form of Ewing's sarcoma (21 cases) consisting of principal blastemic cells and secondary degenerative ones (dark cells) from an atypical variant (7 cases of atypical Ewin's sarcoma) whose structure adopts either an immature mesenchymal or histiocytic appearance. The ultrastructure of the atypical Ewing's cells demonstrates variability in size and in the nucleus, which is grooved and possesses prominent nucleoli. In the cytoplasm, in addition to the existence of a variable amount of glycogen, more highly differentiated structures appear when a comparison is made with the principal cells of the conventional Ewing's sarcoma. There exist filaments and mitochondria complexes, desmosomic-like junctions and profiles of endoplasmic reticulum. No transitional forms between the conventional and the atypical variants of Ewing's sarcoma have been observed. According to this analysis, when compared to other bone malignancies, the presence of glycogen as a means for identifying Ewing's sarcoma seems to be of restricted value. No clinical or anatomical differences could be established between either variant of Ewing's sarcoma.

Topics: Adolescent; Adult; Aged; Bone Neoplasms; Cell Differentiation; Cell Nucleus; Child; Child, Preschool; Cytoskeleton; Desmosomes; Diagnosis, Differential; Female; Glycogen; Humans; Infant; Lymphoma, Large B-Cell, Diffuse; Male; Microscopy, Electron; Middle Aged; Sarcoma, Ewing

Ultrastructural examination of an alveolar rhabdomyosarcoma revealed that the bulk of the tumor was composed of small polygonal cells containing polyribosomes, short strands of rough endoplasmic reticulum, and variable amounts of glycogen; the cells were joined by small desmosome-like structures. Similar cells, as well as a variety of larger forms which appeared to be derived from the small cells, were observed along the septa; the larger forms developed a copious cytoplasm and occasionally produced myofilaments. Some of these cells were separated from the septa by basement membranes. Giant cells were also present in the alveolar spaces; these formed neither basement membranes nor myofilaments. The septa themselves were composed largely of collagen and fibroblasts. It is suggested that the commonly proposed correspondence of alveolar rhabdomyosarcoma to the myotube stage of muscle differentiation is incorrect; if anything, the overall pattern of the tumor is epithelial-like, and may correspond to the organization of mesenchymal cells at the stage of somite differentiation. The resemblance of the masses of small tumor cells to the cells of Ewing's sarcoma is discussed.

Topics: Basement Membrane; Cell Differentiation; Child; Cytoskeleton; Desmosomes; Glycogen; Humans; Male; Microscopy, Electron; Muscles; Rhabdomyosarcoma; Sarcoma, Ewing

The differential diagnosis is difficult in cases of metastatic neuroblastoma, Ewing's sarcoma, lymphoma, and rhabdomyosarcoma, the common so-called small round cell tumors of childhood. The distinction between Ewing's sarcoma and neuroblastoma in bone with no soft tissue mass in the adolescent is especially difficult. Ewing's tumor is usually characterized by its content of glycogen, neuroblastoma by its absence. A case of glycogen-containing neuroblastoma initially misdiagnosed as Ewing's tumor is presented. Diagnostic implications, including the role of electron microscopy in diagnosis, are discussed. Glycogen alone is unreliable as a diagnostic aid due to 1) its presence in several tumors other than Ewing's including neuroblastoma, and 2) its absence in some cases of Ewing's sarcoma.

Topics: Adolescent; Adrenal Gland Neoplasms; Bone Neoplasms; Diagnosis, Differential; Glycogen; Humans; Male; Microscopy, Electron; Neoplasm Metastasis; Neuroblastoma; Sarcoma, Ewing

An autopsy study of 26 cases of Ewing's sarcoma treated with radiation to the primary site plus adjuvant chemotherapy has shown metastatic tumor in 23 cases. Metastases were found typically in lungs, pleura, bones and regional lymph nodes. In three cases no tumor could be found at autopsy, and death was due to complications of treatment. Tumor was found in the irradiated primary site in 13 of the 20 cases in which the primary site was examined at autopsy. Histologically, the tumor at autopsy frequently had increased pleomorphism and increased numbers of bizarre giant cells; however, these changes did not affect the presence of glycogen in tumor cells, thus reaffirming the importance of intracytoplasmic glycogen in the diagnosis of Ewing's sarcoma.

Topics: Adolescent; Adult; Amylases; Bone Neoplasms; Child; Child, Preschool; Female; Glycogen; Humans; Lung Neoplasms; Male; Neoplasm Metastasis; Periodic Acid-Schiff Reaction; Sarcoma, Ewing

Twenty-one cases of Ewing's sarcoma were analyzed by light- and electron-microscopy and the fine structure compared to that described in previous publications. In the predominant "primary" tumor cells, glycogen was abundant in 53% of cases, infrequent in 33%, and rare in 14%. In three cases, microtubules, in association with glycogen, were demonstrated. The so-called differentiated "secondary" reticular tumor cells were sparsely populated in eight cases. Evidence is presented to suggest that these so-called "secondary" reticular cells are merely "primary" tumor cells in a state of regression. Secondary cells and cells with nuclear identations or convolutions were of no discernible prognostic significance. The histogenesis of Ewing's sarcoma remains an enigma but present findings support a primitive mesenchymal origin.

Topics: Adolescent; Adult; Bone Neoplasms; Child; Extremities; Female; Glycogen; Humans; Male; Microtubules; Pelvic Bones; Ribs; Sarcoma, Ewing

The light and the electron microscopic features of an extraskeletal round-cell tumor resembling Ewing's sarcoma are described. Ultrastructural observation revealed features similar to Ewing's sarcoma of bone. Variable agrees of nuclear complexity are described. These soft tissue tumors are probably composed of undifferentiated mesenchymal cells; there is no ultrastructural evidence to indicate the cell of origin.

Topics: Adult; Cell Membrane; Chromatin; Cytoplasmic Granules; Glycogen; Humans; Male; Microscopy, Electron; Mitosis; Organoids; Sarcoma, Ewing; Spinal Cord Neoplasms

Tumour tissue surgically excised from 10 patients bearing Ewing's tumour of bones was examined electron microscopically and histoenzymologically. In all cases the tumour was composed of polygonal cells with cytoplasm poor in organelles but with conspicuous aggregates of glycogen particles. There were numerous intercellular connections of desmosomal type and a distinct cell membrane bound positivity for alkaline phosphatase activity. In two cases in which there was a negative reaction for alkaline phosphatase, the lack of enzyme activity might have been related to cytotoxic treatment carried out for several months immediately before excision of the tissue used for histoenzymological studies. The problem of histogenesis of Ewing's tumour remains unresolved although some of the present findings support a haemangiogenic origin of the tumour.

Topics: Adolescent; Adult; Alkaline Phosphatase; Bone Neoplasms; Cell Membrane; Female; Glycogen; Humans; Intercellular Junctions; Male; Microscopy, Electron; Sarcoma, Ewing

In addition to a light-microscopical and histochemical investigation of primary and metastic lesions in 27 cases of Ewing's sarcoma, biopsy materials from the primary bone lesions of 7 patients with this neoplasm were examined histochemically, enzyme-cytochemically and electron microscopically to elucidate the histogenesis and nature of the neoplasm. Ultrastructural observation has revealed that besides intracytoplasmic and extracellular deposition of glycogen the tumor cells possess several cytological features characterized by intracytoplasmic microfilaments of varying thickness up to 80 A, occasional appearance of dense patches, fat droplets, desmosomal connections and reminiscent attachment bodies. The tumor cells are mostly round, oval or polygonal in shape, but spindle or elongated cells are intermingled and occasionally contain well-developed rough endoplasmic reticulum, resembling pericytes or fibroblasts. In the intercellular spaces amongst the tumor cells, varying amounts of variable-shaped amorphous materials are found, which are compatible with acid mucopolysaccharides and glycoproteins histochemically verified. These findings may suggest that Ewing's sarcoma is a highly malignant neoplasm originating from a transitional cell developed from pericytes to vascular smooth muscle cells in the bone marrow.

Topics: Acid Phosphatase; Adenosine Triphosphatases; Alkaline Phosphatase; Bone Neoplasms; Glycogen; Humans; Sarcoma, Ewing

Cytologic and cytochemical examination of eighteen cases of round-cell sarcoma of bone allowed classification of these tumors into four cytologic groups. Additional cytochemical examinations based on the PAS and D-PAS reactions, and the demonstration of the activity of peroxidase, naphtol-ASD-Chloracetate esterase, alpha-naphthylacetate esterase, naphthol-AS-acetate esterase with and without sodium fluoride inhibition, acid and alkaline phosphatases yielded no evidence of uniform behavior among the individual groups or within any single group. The studies showed that a positive glycogen reaction cannot be used as a basic criterion for the classification of such tumors as Ewing's sarcoma and for regarding them as a uniform tumor group. It is possible that a pool of tumors is involved, including tumors of monocytic and probably of lymphocytic origin, reticulum-cell sarcoma, tumors of myelocytic and erythroplastic origin, stem-cell tumors, and endothelial-cell tumors. Histologic examination alone is not sufficient for the classification of round-cell sarcomas of bone, and it should be supplemented by cytologic and cytochemical or histochemical methods. Osteosarcomas (23 cases) and chondrosarcomas (8 cases) display cells which are characteristic for these tumors and which could be correlated with their benign counterparts, osteoblasts and chondroid cells. The histologically recognizable degree of malignancy of chondrosarcoma can be evaluated better with the cytologic than with the histologic technic. Indications of the possibilities of differential diagnosis based on the cytologic pictures of benign and malignant osteoplastic and chondroplastic tumors, giant-cell tumors and chordoma are discussed.

Topics: Adolescent; Bone Neoplasms; Cell Nucleolus; Cell Nucleus; Child; Chondroblastoma; Chondroma; Chondrosarcoma; Chordoma; Cytoplasm; Female; Giant Cell Tumors; Glycogen; Humans; Infant; Male; Naphthol AS D Esterase; Osteosarcoma; Periodic Acid-Schiff Reaction; Phosphoric Monoester Hydrolases; Sarcoma; Sarcoma, Ewing

Topics: Adolescent; Adult; Biopsy; Bone Neoplasms; Child; Culture Techniques; Female; Glycogen; Hemangioma; Humans; Lymphoma, Large B-Cell, Diffuse; Male; Microscopy, Electron; Sarcoma, Ewing

Topics: Adolescent; Adrenal Gland Neoplasms; Aged; Bone Neoplasms; Catecholamines; Child; Child, Preschool; Dactinomycin; Female; Fibula; Glycogen; Humans; Humerus; Lymphoma, Large B-Cell, Diffuse; Lymphoma, Non-Hodgkin; Male; Microscopy, Electron; Middle Aged; Neuroblastoma; Pelvic Neoplasms; Reticulin; Sarcoma, Ewing; Staining and Labeling; Sternum; Vincristine

Topics: Alkaline Phosphatase; Biopsy; Bone Cysts; Bone Diseases; Bone Neoplasms; Chondroma; Chondrosarcoma; Clinical Enzyme Tests; Diagnosis, Differential; Femoral Neoplasms; Fibrosarcoma; Giant Cell Tumors; Glycogen; Hemangioma; Hemangiosarcoma; Histocytochemistry; Humans; Ilium; Lymphoma, Large B-Cell, Diffuse; Multiple Myeloma; Neoplasm Metastasis; Osteoma; Osteoma, Osteoid; Osteomyelitis; Osteosarcoma; Ribs; Sarcoma, Ewing; Tibia; Trephining; Tuberculosis, Osteoarticular

Topics: Autopsy; Biopsy; Bone Neoplasms; Child; Female; Glycogen; Histocytochemistry; Humans; Lumbar Vertebrae; Microscopy, Electron; Phagocytosis; Radiography; Sarcoma, Ewing

Topics: Bone and Bones; Diagnosis, Differential; Glycogen; Humans; Lymphoma, Large B-Cell, Diffuse; Lymphoma, Non-Hodgkin; Neoplasms; Sarcoma; Sarcoma, Ewing