SUPEROXIDE DISMUTASE IN JUVENILE NEURONAL CEROID-LIPOFUSCINOSIS (SPIELMEYER-VOGT-BATTEN'S DISEASE)

Abstract— The ceroid-lipofuscin pigments found in neuronal ceroid-lipofuscinosis are believed to arise from lipid peroxidation-damaged organelle membranes. In several in vitro systems superoxide dismutase inhibits lipid peroxidation. Erythrocytes and lymphocytes were prepared from ten patients with juvenile neuronal ceroid-lipofuscinosis (Spielmeyer-Vogt-Batten's disease). Copper-zinc and manganese superoxide dismutases were selectively assayed by a direct method on homogenates of the cells. No significant differences were found between the patients, neurological controls and healthy adults.     

Methylated lysine in storage body protein of sheep with hereditary ceroid-lipofuscinosis.

Juvenile ceroid-lipofuscinosis (Batten disease) is a hereditary storage disease with an autosomal-recessive mode of transmission. This disorder has been identified in humans, dogs and sheep. It is characterized by massive accumulations of autofluorescent storage bodies in many tissues. This storage body accumulation is accompanied by functional decline and degeneration of the affected tissues, and ultimately results in premature death. The primary defect responsible for juvenile ceroid-lipofuscinosis has not been identified. Previous studies have indicated that the storage material is primarily protein. Why this protein accumulates in storage bodies remains to he determined. In affected humans, the storage body protein appears to be abnormally rich in a methylated derivative of lysine (epsilon-N-trimethyllysine). Studies were undertaken to determine whether the storage bodies from sheep with hereditary ceroid-lipofuscinosis were also characterized by the presence of this modified amino acid. Chromatographic and mass spectral analyses of hydrolysates of the storage body protein indicated a significant fraction of the lysine residues in this protein were present as the epsilon-N-trimethyl derivative. This modified amino acid was not detected in hydrolysates of protein from normal sheep tissues or from tissues of sheep with GM1 gangliosidosis, nor did it appear to be present in the storage body protein from a human subject with the late infantile form of ceroid-lipofuscinosis. Thus, it is apparently specific to the storage body protein that accumulates in the juvenile type of this disease. The abnormal presence of epsilon-N-trimethyllysine in proteins could interfere with their sorting or degradation within cells and thus cause them to accumulate in the storage bodies characteristic of the human juvenile and ovine ceroid-lipofuscinoses.     

Analysis of dolichyl pyrophosphoryl oligosaccharides in purified storage cytosomes from ovine ceroid-lipofuscinosis

Ovine ceroid-lipofuscinosis is an inherited neurodegenerative disorder characterised by the accumulation of storage cytosomes in brain and visceral organs. Phosphorylated dolichol-containing compounds, largely in the form of dolichyl pyrophosphoryl oligosaccharides, have been shown to constitute 1-2% of the dry weight of storage cytosomes isolated from brain and pancreas, and 0.5 and 0.1% respectively of storage cytosomes isolated from liver and kidney. The carbohydrate portion of these glyconjugates in storage cytosomes isolated from brain, pancreas and liver consisted of a series of oligosaccharides of composition Man2-9GlcNAc2, with Man5-8GlcNAc2 predominating. The concentrations of dolichyl pyrophosphoryl oligosaccharides in storage cytosomes from ovine ceroid-lipofuscinosis are much higher than has been reported for endoplasmic reticulum, their normal functional location.     

LEUKOCYTE PPD-PEROXIDASE ACTIVITY WITH POLYUNSATURATED FATTY ACID HYDROPEROXIDES: NORMAL VALUES IN BATTEN''S DISEASE

Abstract— The activity of leukocyte p -phenylenediamine (PPD)-dependent peroxidase with respect to 3 peroxidic substrates was investigated in three patients with Batten's disease (ceroid-lipofuscinosis) as compared with normal controls: 1. The activity of PPD-peroxidase, using H₂O₂ as the peroxidic substrate, was found to be normal in our patients with Batten's disease. 2. PPD-peroxidase was shown to be active towards arachidonic acid hydroperoxide (AAHPO) and linoleic acid hydroperoxide (LAHPO) as peroxidic substrates. No difference could be detected between patients and normals. 3. Determination of Michaelis constants with respect to H₂O₂, AAHPO and LAHPO in normal leukocytes revealed that PPD-peroxidase was more active towards AAHPO (lower K _m) than towards LAHPO. The same kinetic properties were found for PPD-peroxidase in patients with ceroid-lipofuscinosis.     

Ultrastructural study in the diagnosis of various neurolipidoses

We have studied the sphingolipidosis, principally, ceroid-lipofuscinosis and other complex lipid storage diseases. We examined biopsy tissue of fourteen patients with diseases of the various types given above. The biopsy tissue was usually skin, muscle or peripheral nerve, and occassionally brain or various visceral tissue.     

Significance of histiocytes on otherwise-normal cervical smears from postmenopausal women. A retrospective study of 108 cases

OBJECTIVE: To evaluate the significance of histiocytes on normal cervical smears from postmenopausal women and correlate them with endometrial pathology. STUDY DESIGN: Histiocytes were classified into three types. The clinical history was obtained from cytologic and surgical reports. RESULTS: Among 108 cervical smears, 13 had large, foamy histiocytes (type A), 88 had histiocytes resembling superficial endometrial stromal cells (type B), and 7 had variably sized histiocytes alone or in association with inflammatory or multinucleated cells (type C). Endometrial pathology was identified in 13 patients (12.0%): 4/13 with type A histiocytes (2 endometrial adenocarcinomas, 2 endometrial polyps), 8/88 with type B histiocytes (8 endometrial polyps) and 1/7 with type C histiocytes (endometrial polyp). Among 70 patients with no clinical indications for endometrial sampling except for the presence of histiocytes, 4 demonstrated endometrial pathology (all endometrial polyps). In contrast, endometrial pathology was identified in 9/38 with clinical indications for endometrial sampling. Among the 13 patients with endometrial pathology, 9 had a significant clinical history (sensitivity of 69.2%), and 4 had histiocytes as the only indication for endometrial biopsy (sensitivity of 30.8%). CONCLUSION: A significant clinical history is more predictive of endometrial pathology and outweighs the significance of histiocytes as an indication for endometrial biopsy.     

Crush cytology of Rosai-Dorfman disease of the central nervous system. A report of 2 cases

BACKGROUND: Rosai-Dorfman disease (RDD) of the central nervous system (CNS) is rare. There is no previous report on intraoperative crush cytology of this disorder. CASES: A 70-year-old man presented with headache and was found to have 2 intracranial extraaxial masses. A 62-year-old man presented with cauda equina syndrome secondary to a mass lesion in the sacral canal. The crush smears, prepared intraoperatively from a small portion of the biopsy specimen in each case, showed scattered lymphoid aggregates in a lighter background with a loose admixture of lymphocytes, plasma cells, usual histiocytes and RDD histiocytes. The RDD histiocytes contained single but sometimes multiple nuclei that were larger and more hyperchromatic than those of the usual histiocytes. Many RDD histiocytes had voluminous, pale pink cytoplasm. Lymphophagocytosis by RDD histiocytes was observed. The findings in the crush smears reflected the histology of the lesions. The lymphophagocytosis and nuclear details in the RDD histiocytes were more visible in the crush smears than in the frozen sections. CONCLUSION: Crush cytology appears useful, alone or in conjunction with frozen sections, in the intraoperative diagnosis of CNS RDD.     

Dolichyl-pyrophosphoryloligosaccharide protein oligosaccharide transferase in neuronal ceroid-lipofuscinosis.

The kinetics of the oligosaccharide transfer from oligosaccharyl pyrophosphoryldolichol to endogenous protein acceptors in human fibroblasts were studied. No alterations in the transferase activity and enzyme characteristics could be observed in fibroblasts from neuronal ceroid-lipofuscinosis (NCL) patients. Analysis of urinary dolichol of two NCL patients also did not reveal substantial differences with respect to controls.     

Neuronal ceroid-lipofuscin storage in a cynomolgus monkey (Macaca fascicularis)

The light and electron microscopical characteristics of non-age-related neuronal ceroid-lipofuscinosis in a young cynomolgus monkey (Macaca fascicularis) are described. The pigment in the neuronal cytoplasm was PAS-positive, sudanophilic, weakly acid-fast, and positive with Schmorl's and aldehydefuchsin stains for lipofuscin. Ultra-structurally, it appeared as membrane-bound, electron-dense aggregates within the cytoplasm of neurones of both brain and spinal cord.     

Distribution pattern of concanavalin A on carcinoma cells, histiocytes and mesothelial cells from effusions

Fixed and unfixed cancer cells, mesothelial cells and histiocytes were exposed to fluorescein-labeled concanavalin A (ConA-FITC). Cancer cells, whether fixed or unfixed, showed a similar pattern of fluorescence, as a continuous layer over the whole periphery of the cell. This pattern of ConA-FITC distribution was also obtained on fixed mesothelial cells and fixed histiocytes. Redistribution of ConA-FITC in form of "caps" and "patches" was recorded on unfixed mesothelial cells and unfixed histiocytes. Of the three cell types studied, only the histiocytes were lysed by the incubation with ConA-FITC.     

Cytologic diagnosis of hemophagocytic lymphohistiocytosis in ascitic fluid: a case report

BACKGROUND: Hemophagocytic lymphohistiocytosis (HLH), also known as macrophage activation syndrome, is a rare, fatal hematopoietic disease. Its cytologic features may be subtle because the abnormal histiocytes may not be recognized if one is not aware of this entity. We report a case of HLH involving the ascitic fluid. CASE REPORT: A 73-year-old man developed weakness, lethargy, decreased appetite and progressive shortness of breath after a cholecystectomy. Physical examination revealed hypotension, tachycardia and chest dullness with decreased breath sounds bilaterally. Radiologic examination revealed bilateral pleural effusions. The patient accumulated fluid in the peritoneal cavity, lungs, retroperitoneum and mediastinum. Bone marrow biopsy showed abundant histiocytes infiltrating the marrow cavity, and many of these histiocytes contained cellular debris. A diagnosis of HLH was therefore made. The abdominal paracentesis specimen contained many similar histiocytes exhibiting erythrophagocytosis and lymphophagocytosis. These abnormal histiocytes were positive for CD68 and negative for AE1/AE3, confirming the diagnosis of HLH. The patient died soon after from disseminated aspergillosis. CONCLUSION: HLH is cytologically characterized by the presence of abnormal histiocytes with ingested cellular debris. In serous effusions they should not be confused with mesothelial cells. Immunohistochemical studies may help confirm the diagnosis.     

Infantile neuronal ceroid-lipofuscinosis is not an allelic form of Batten disease: exclusion of chromosome 16 region with linkage analyses

Infantile neuronal ceroid-lipofuscinosis (CLN1) is the form of neuronal ceroid-lipofuscinoses (NCL) with the earliest onset of symptoms. The locus of the most common form of these disorders, juvenile NCL (CLN3), has been mapped to chromosome 16. We report here linkage data of the same region in Finnish CLN1 families. Our results indicate that CLN1 is not allelic with CLN3 but represents a different locus, which is not located within about 70 cM in chromosome 16.     

Value of histiocyte detection in Pap smears for predicting endometrial pathology. An institutional experience

OBJECTIVE: To determine the clinical implications of the finding of histiocytes in Pap smears in 1 patient population. STUDY DESIGN: The medical records and Pap smears which the presence of histiocytes was mentioned in the diagnosis between August 1996 and August 2001 were reviewed in conjunction with follow-up surgical findings. The positive predictive value (PPV) for significant endometrial pathology for the isolated finding of histiocytes on Pap smear was determined. RESULTS: Of the 238,225 women screened over a 60-month period, 325 were reported to have histiocytes in their Pap smears. Of them, 238 (73.2%) had subsequent endometrial sampling, hysterectomy or both, and follow-up Pap smears. Two hundred seven smears (87%) failed to disclose endometrial pathology. Thirty-one cases (13%) resulted in significant histopathologic findings, including 12 uterine malignancies, 8 endocervical polyps, 7 endometrial polyps, 2 submucosal leiomyomata, 1 simple hyperplasia without atypia and 1 case of tamoxifen-related changes. Upon review of the clinical records, 58% (18/31) of those patients had other significant clinical and/or cytologic findings. Five of the 18 patients (27.8%) had associated postmenopausal bleeding, 11 had additional abnormal Pap smear findings (atypical glandular cells, 6/18, or 33.3%; endometrial cells, 5/18, or 27.8%), and another 2 had both postmenopausal bleeding and atypical glandular cells (2/18, or 11.1%). The PPV for significant uterine pathology for women with the isolated finding of histiocytes on a Pap smear was 5.5% and 60% with additional clinical and/or Pap smear findings. The PPV for endometrial cancer was 1.3% in women with the isolated finding of histiocytes on a Pap smear but 20% for women with histiocytes and additional clinical/or Pap smear findings. CONCLUSION: Based on the findings of this study and recently published data, we conclude that the isolated finding of increased histiocytes in the absence of postmenopausal bleeding, endometrial cells or atypical glandular cells on a Pap smear is a poor indicator of uterine disease.     

Histiocytes and the detection of endometrial adenocarcinoma

Histiocytes have long been recognized as part of the milieu of endometrial carcinoma in gynecologic smears. In an effort to determine whether a quantitative assessment of histiocytes, especially in the absence of endometrial cells, could increase the effectiveness of the cervicovaginal smear for diagnosis of endometrial carcinoma, smears obtained prior to a tissue diagnosis of endometrial adenocarcinoma were evaluated from 44 postmenopausal women. Smears from 97 age-matched patients in the same clinic were also evaluated and used as a control group for the endometrial carcinoma patients. All smears were evaluated for the presence of histiocytes and for the presence of benign or malignant endometrial cells, with the histiocytes quantitated as minimal (less than 5 per high-power field [HPF]), moderate (5 to 10/HPF) or heavy (greater than 10/HPF). Sensitivity and specificity were calculated to assess the role of histiocytes in the presence and in the absence of endometrial cells using cytologic findings. Our data indicate that the presence of moderate or heavy numbers of histiocytes on cervicovaginal smears of postmenopausal women increased the cytologic sensitivity from 61% to 82% when considered a marker of disease along with endometrial cells. These results suggest that attention to the presence of histiocytes on cervicovaginal smears may increase the utility of cytology for the diagnosis of endometrial lesions and may be a useful guideline for the cancer-related gynecologic examination.     

Identification of tissue histiocytes on paraffin sections by a new monoclonal antibody

A mouse monoclonal antibody (MAC 387) with specificity for monocytes and tissue histiocytes was produced by immunization of a BALB/c mouse with peripheral blood monocyte components derived by affinity chromatography of detergent-solubilized monocyte material on Sepharose 4B coupled to rabbit anti-monocyte antibodies. MAC 387 strongly stained the cytoplasm of cells of the monocyte/macrophage series on paraffin sections after controlled trypsinization of sections. The antibody showed broad reactivity for a variety of tissue histiocytes, including infiltrating and reactive histiocytes, alveolar macrophages, Kupffer cells, follicle-center macrophages, splenic red pulp macrophages, tumor-infiltrating macrophages, sinus histiocytes, epithelioid giant cells (variably), and cases of histiocytosis X and dermatopathic lymphadenopathy. Molecular weight data obtained by Western blotting, immunoprecipitation, and immunoaffinity-purification revealed that the antigen was present in different forms in the monocyte and granulocyte. In the granulocyte, free alpha (Mr 12 KD) and beta (Mr 14 KD) chains expressing the MAC 387 epitope were found together with associations of one alpha and one beta chain linked by disulfide bonds to yield a heterodimer of Mr 26 KD. In the monocyte, free alpha and beta chains are not found, but instead the heterodimer and associations of two (Mr 56 KD) and four (Mr 112 KD) heterodimers are disulfide-linked together. This new monoclonal reagent should have particular value for identification of tissue histiocytes in routine paraffin sections and particularly for demonstration of histiocytes in malignant lymphomas.     

Adhérence de granulocytes,utilisés in vivo lors d'une stimulation repetée,sur des cellules histiocytaires issues de l'épiploon de souris cultivé in vitro

Summary Mice were immunized by repeated intraperitoneal injections of pig granulocytes, at 48 hour intervals. Animals were sacrificed 18 hours after the last injection. Stimulated and non-stimulated omenta were cultivated for various times up to 18 days. In the highly developed population of cells migrating from the stimulated explants, histiocytes with a large, extended surface appeared by the 6th day of growth. When cultures were incubated in vitro with the antigen, the histiocytes were capable of fixing granulocytes on their surface. This phenomenon was especially displayed by the large histiocytes. In control cultures, resulting from non-stimulated omenta, adherent granulocytes were only occasionally observed and no large histiocytes appeared. — The significance of these figures of adherence with respect to the immune treatment is discussed.

     

Pleural effusion cytology in a case of cytophagic histiocytic panniculitis (subcutaneous panniculitic T-cell lymphoma). A case report

BACKGROUND: Cytophagic histiocytic panniculitis (CHP) presents with subcutaneous panniculitis associated with hemophagocytic syndrome. Many cases of CHP are now being classified as a natural disease progression of subcutaneous panniculitic T-cell lymphoma (SPTL). There have been no cytologic reports dealing with pleural aspirates in cases of CHP or SPTL. CASE: A pleural aspirate obtained from a 19-year-old female revealed lymphoma cells and hemophagocytic histiocytes. A skin biopsy specimen showed the presence of CD8-positive lymphoma cells in fat lobules associated with cytologically benign histiocytes with erythrophagocytosis and lymphophagocytosis. CONCLUSION: Hemophagocytic histiocytes were seen in the pleural effusion from a patient with SPTL.     

Fine needle aspiration cytologic findings in a case of sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman syndrome)

The clinical, fine needle aspiration (FNA) cytologic and histologic findings in a case of sinus histiocytosis with massive lymphadenopathy (SHML) in a five-year-old boy are presented. The FNA smears showed a proliferation of plasma cells, lymphoid cells and histiocytes demonstrating emperipolesis that is characteristic of this benign disorder; these cytologic findings were reported as a reactive lymphoid infiltrate with a high content of benign histiocytes. The diagnosis was made on a subsequent open biopsy performed on the persistent lymphadenopathy. The differential diagnosis of lymph nodes containing a prominent component of histiocytes is discussed.     

Histiocyte X positivity for nonspecific esterase

The cell membrane and X bodies of histiocytes X from two cases of eosinophilic granuloma were stained by nonspecific esterase. The results show that histiocytes X possess a cell membrane exoenzyme similar to that of other histiocytes, such as alveolar macrophages, but the role of the X body as related to enzyme activity remains an unknown.     

Effect of Temperature on the Intracellular Growth of Histoplasma capsulatum

The rate and form of growth of Histoplasma capsulatum within histiocytes derived from homothermic and poikilothermic animals, and incubated at 25, 30, and 37 C, are described. The generation time of the fungus in mouse cells incubated at 37 and 25 C was 11 and 24 hr, respectively. Blastospore formation was progressively retarded in cells at 25 C, and this retardation was accompanied by germination of some of the blastospores. The generation time of the fungus in mouse cells incubated at 30 C was the same as it was at 37 C. Germ tube formation was not a prominent feature of intracellular growth at 30 C. The rate of growth of H. capsulatum within frog histiocytes at 30 and 25 C was slower than it was in mouse cells at the same temperatures. Some loss of frog histiocytes in cultures incubated at 37 C prevented accurate estimation of the rate of growth of the fungus at this temperature. Growth of H. capsulatum in frog histiocytes kept at 25 C was progressively retarded, and the retardation was accompanied by germination of the yeasts. Yeast-phase growth predominated in fish histiocytes incubated at 30 C, whereas germ tubes were formed within such cells incubated at 25 C. Cell survival of fish histiocytes was relatively poor in culture, and no estimates of rate of growth of the fungus within these cells were made.