SPHINGOLIPIDS AND PHOSPHOLIPIDS IN MICROSOMES AND MYELIN FROM NORMAL AND PATHOLOGICAL BRAINS

—Myelin and microsomes were separated from human cerebral white matter and cortex respectively using the technique of 15% caesium chloride and their sphingolipid and phospholipid contents estimated. Normal brains as well as cerebral material from cases of metachromatic leucodystrophy, Krabbe's disease and Tay-Sachs’disease were studied. Gangliosides were not present in normal myelin but were found in microsomes and in myelin from the pathological material. The ratio of cerebroside to sulphatide in myelin was 4 to 1 in normal, 1 to 20 in metachromatic and 7 to 1 in Krabbe's disease. The results in the human material are briefly discussed.     

Plasma corticotrophin levels in addison-schilder's disease.

Raised plasma immunoreactive corticotrophin (ACTH) levels were found in five boys with the sex-linked disorder progressive leucodystrophy associated with adrenal insufficiency (Addison-Schilder''s disease) and in a symptom-free brother of one of them. Similar ACTH concentrations were found using two antisera, one against the N-terminal part of the ACTH molecule and the other against the C-terminal part. In one patient the circulating ACTH had normal biological activity as measured using the cytochemical ACTH bioassay. Immunoreactive beta/-melanocyte-stimulating hormone was also determined in one patient and found to be raised.     

Nephrotic Syndrome in Chronic Lymphocytic Leukaemia

Two patients with chronic lymphocytic leukaemia and the nephrotic syndrome are described in whom deposits were shown in renal glomerular basement membranes in a pattern suggesting immune-complex glomerulonephritis. This renal lesion has been described in one case of squamous carcinoma of the bronchus, in one case of Burkitt''s lymphoma, and in three cases of Hodgkin''s disease though not previously in chronic lymphocytic leukaemia. Immune-complex glomerulonephritis is, however, a recognized finding in mice infected with leukaemogenic viruses     

Occurrence of brain calcifications,iron deposits,and tonic-clonic convulsions following experimental infections with Candida albicans

Summary During the prolonged presence ofCandida albicans in the spleen of Wistar rats, two cases of calcium deposits in the brain were observed. In one case there was one big calcium deposit at some distance from the cortex. In this case, thickening in a limited area of the leptomeninges was found. The animal had generalized tonicclonic convulsions which lead to death. In the second case, calcium deposits formed two separate agglomerations in the posterior part of the thalamus. Serial sections made at six micron intervals showed the presence of calcium deposits to a depth of forty-eight microns. At twenty-four microns other calcium deposits were observed in the thalamus symmetrically located to those described above.Small iron deposits were found in the anterior part of the thalamus and other iron deposits were found in the temporal lobe of the brain. They were arranged in a straight line along the capillary vessels and most of them were seen in the macrophages in the vicinity of the vessels.Since calcium deposits in the brain occurred in the same experimental conditions as reported cases of calcinosis, it is probable that the same mechanism is involved in all these pathological calcium deposits.This work was supported by a Damon Runyon Memorial Fund Grant Nr. 720.     

STRUCTURAL STUDIES ON SULPHATIDES IN METACHROMATIC LEUCODYSTROPHY

Sulphatides have been isolated by a newly developed method from the brain white matter of normal human adult and child, and of two patients with metachromatic leucodystrophy. Only galactose was found in chromatographic analysis of the carbohydrate component. Following permethylation of the sulphatides, the methylated hexose, in all cases, was 2,4,6-trimethylgalactose. The sulphate group is located solely at the C-3 position of the galactopyranose moiety, and in metachromatic leucodystrophy, although there is an accumulation of sulphatides, their composition and structure remain normal.     

Hereditary demyelinating diseases - the leukodystrophies

Schilder's disease and Greenfield's metachromatic leucodystrophy are hereditary demyelinating diseases, which are probably metabolic and lysosomal, although the responsible enzyme is not yet known for all.     

Absence of an orphan mitochondrial protein, c19orf12, causes a distinct clinical subtype of neurodegeneration with brain iron accumulation

The disease classification neurodegeneration with brain iron accumulation (NBIA) comprises a clinically and genetically heterogeneous group of progressive neurodegenerative disorders characterized by brain iron deposits in the basal ganglia. For about half of the cases, the molecular basis is currently unknown. We used homozygosity mapping followed by candidate gene sequencing to identify a homozygous 11 bp deletion in the orphan gene C19orf12. Mutation screening of 23 ideopathic NBIA index cases revealed two mutated alleles in 18 of them, and one loss-of-function mutation is the most prevalent. We also identified compound heterozygous missense mutations in a case initially diagnosed with Parkinson disease at age 49. Psychiatric signs, optic atrophy, and motor axonal neuropathy were common findings. Compared to the most prevalent NBIA subtype, pantothenate kinase associated neurodegeneration (PKAN), individuals with two C19orf12 mutations were older at age of onset and the disease progressed more slowly. A polyclonal antibody against the predicted membrane spanning protein showed a mitochondrial localization. A histopathological examination in a single autopsy case detected Lewy bodies, tangles, spheroids, and tau pathology. The mitochondrial localization together with the immunohistopathological findings suggests a pathomechanistic overlap with common forms of neurodegenerative disorders.     

Role of fine needle aspiration cytology in evaluation of paediatric lymphadenopathy

Fine needle aspiration cytology (FNAC) in children has been slow to gain acceptance as compared with its use in adults. The aim of this retrospective study is to determine the utility of FNAC in diagnosing paediatric lymphadenopathy. Six hundred and ninety-two paediatric lymph nodes were aspirated during the period from January 1996 to June 2001. Aspirated material in 32 cases (4.6%) was deemed inadequate for cytological diagnosis. In the remaining cases, 62.2% were reactive lymphadenopathy, 25.2% tuberculous lymphadenitis, 6.3% acute suppurative lymphadenitis, nine cases revealed malignancy (seven lymphoma, two metastatic deposits) and one case had Langerhan's cell histiocytosis. No significant complications were encountered in any of the cases. FNAC emerged as a safe and reliable diagnostic procedure in paediatric age group obviating the need for excision biopsy.     

IgA nephropathy in adults: immunohistologic findings and clinical course

Laboratory examination of specimens from 123 consecutive renal biopsies performed at Victoria General Hospital, Halifax revealed six cases of mesangial deposition, predominantly of IgA, unassociated with systemic disorders. Immunohistologic examination showed deposits of only IgA in one specimen, IgA and IgG in two and IgA, IgG and IgM in three. Glomerular deposits of C3 were seen in five of the specimens, and properdin was seen in three. Glomeruli in all the specimens showed increased matrix and increased numbers of cells in the mesangium. Electron microscopy revealed deposits in the mesangium or capillary wall in all five of the specimens so studied. All six patients had proteinuria, four had microscopic hematuria, and three had hypertension; in one patient the disease progressed to renal failure.     

Invasive meningococcal disease associated with a very high case fatality rate in the North-West of Poland

The aim of the study was to investigate invasive meningococcal disease in the North-West of Poland, associated with a case fatality rate of 42.9%, where among the first 11 cases, eight had fatal outcome. All fatal cases were diagnosed as fulminant meningococcal severe sepsis with Waterhouse-Friderichsen syndrome and disseminated intravascular coagulation. Serotyping and pulsed-field gel electrophoresis analysis revealed that the high case fatality rate was not associated with the dissemination of one epidemic clone. However, six cases, all with good outcomes, were caused by C:2b:(P1.2,P1.5) isolates of the same pulsed-field gel electrophoresis type belonging to ST8 complex/Cluster A4.     

Reduced Proteins in Temporal Cortex in Alzheimer''s Disease: An Electrophoretic Study

Cytoplasmic and pellet fractions from post-mortem temporal cortex from eight cases of neuropathologically confirmed Alzheimer's disease, one case of cerebrovascular dementia, and five controls were examined by sodium dodecylsulphate-polyacrylamide gel electrophoresis. No differences were observed in the cytoplasmic proteins from the five controls and the case of cerebrovascular dementia. In five cases of Alzheimer's disease with neurone loss, there was a major loss of a cytoplasmic 55,000-dalton protein identified as tubulin and variable reductions in cytoplasmic proteins of molecular weights of 28,000, 30,000, 92,000, and 200,000 daltons. Three cases of Alzheimer's disease had no detectable neurone loss; two of these cases had protein patterns indistinguishable from the controls and one showed some reduction in soluble tubulin only. These results indicate that decreases of particular proteins in the temporal cortex in Alzheimer's disease may be associated with neurone loss.     

THE PATTERNS OF ARYLSULPHATASES A AND B IN HUMAN NORMAL AND METACHROMATIC LEUCODYSTROPHY TISSUES AND THEIR RELATIONSHIP TO THE CEREBROSIDE SULPHATASE ACTIVITY

Abstract— The arylsulphatase A and B patterns of human tissues and leucocytes have been established by isoelectric focussing. Assay conditions, which enable an evaluation of these patterns as quantitatively as possible, have been studied. The dependences of the enzyme patterns on the origin of the tissues and on the storage conditions have been determined. The arylsulphatase A obtained by isoelectric focussing exhibits cerebroside sulphatase activity in the presence of detergents. A purified preparation of the arylsulphatase B likewise shows a significant, although low, cerebroside sulphatase activity. In cases of the conventional types of metachromatic leucodystrophy the arylsulphatase A activity is missing, while in an atypical form of this disease ('ML Variant' according to A ustin et al . (1965) the arylsulphatase A, B and C activities are deficient. In both forms, however, residual activities of the deficient enzymes could be detected which showed isoelectric points identical to those of the normal enzymes.
The following nomenclature is proposed: 'Variant B' for the conventional type, in which the arylsulphatase B activity is present, and 'Variant O' for the exceptional cases, in which all arylsulphatase activities are deficient. The significance of the cerebroside sulphatase activity of arylsulphatase B for a possible residual turnover of cerebroside sulphates in the conventional type of the disease is discussed.     

Fine structure of in situ microbial iron deposits

The fine structure of iron deposits produced by Leptothrix spp. from two caverns and a surface spring were observed. In the case of Leptothrix pseudovacuolata, the iron occurred as an amorphous deposit within the cell sheath and the cells contained gas vesicles. In the case of the other Leptothrix spp., the iron was deposited within the cell sheath either as a hexagonal or fibrillar matrix. In the case of fibrillar iron deposits, fibrils resembling those of the sheath were found within the cytoplasm of the cells. This suggests that the deposition of iron could have occurred within the cells as well as at the cell surface. In some cases, the fine structure of microbial iron deposits could provide a means of distinguishing biological from abiological iron deposition.     

Removal of glomerular deposits induced by either preformed immune complexes or by a chronic immune complex model in NZB/W mice

The solubilization and removal of defined glomerular immune complex deposits by excess antigen was examined in NZB/W female mice. Glomerular deposits were induced by administering preformed immune complexes to young (2 to 4 mo) mice before they naturally acquired deposits from endogenous disease and to old (7 mo) mice with deposits from naturally acquired disease. The administration of excess antigen specifically removed deposits of preformed immune complexes in both groups. This was associated with a reduction in circulating large latticed complexes containing more than two antigen and two antibody molecules (greater than Ag2Ab2). Established deposits in old mice therefore did not interfere with removal of newly induced deposits of preformed immune complexes. Glomerular deposits were also induced in young mice by a chronic human serum albumin (HSA) immune complex model. The antigen in immune deposits induced by 2 wk of chronic antigen administration was solubilized and was removed within 48 hr of administering excess antigen. Circulating antibodies to the antigen were also reduced by excess antigen. Glomerular deposits of mouse immunoglobulin and complement were not significantly reduced by excess antigen but remained more intense than in mice of comparable age given preformed complexes. Thus deposits of other antigen antibody systems and possibly endogenous disease were induced by the chronic HSA immune complex model in NZB/W mice. However, defined antigen deposits within deposits containing multiple antigen antibody systems can clearly be removed by administering excess antigen.     

Pathenogenesis and Therapy of Liver Disease in Protoporphyria

Protoporphyria is an inherited disorder in man characterized by the overproduction of protoporphyrin, a compound that is excreted by the liver. Hepatobiliary disease may occur in protoporphyria, and several cases have been reported in which death was due to liver disease. Based on the histological evaluation of liver biopsy specimens from 18 patients, 6 of whom died with cirrhosis and liver failure, we speculate that liver disease in this condition is caused by protoporphyrin deposits in hepatobiliary structures. These deposits are composed of crystals and have a characteristic birefringence when examined by polarization microscopy.One patient with early liver damage was given cholestyramine and vitamin E in an attempt to reduce the amount of protoporphyrin which the liver excreted daily. Liver function tests returned to normal, and red cell and plasma protoporphyrin levels decreased. A repeat liver biopsy after one year of therapy showed healing, with decrease of the protoporphyrin deposits.Liver disease in protoporphyria may be treated by directing therapy toward the metabolic abnormality.     

Evaluation of abdominal fat pad aspiration cytology and grading for detection in systemic amyloidosis

OBJECTIVE: To evaluate the diagnostic efficacy of abdominal fat pad aspiration cytology as a screening procedure for systemic amyloidosis and to assess the clinical usefulness of semiquantitative grading criteria of fat pad amyloid deposits. STUDY DESIGN: Aspiration cytology samples from 297 cases of abdominal fat pad were retrospectively analyzed for amyloid deposits. The smears were graded semiquantitatively. The deposits in the smears were compared with histologic evidence of amyloidosis in deeper tissues in 44 cases. RESULTS: Retrospective analysis of 297 cases of aspiration cytology revealed amyloid in 90 cases. Follow-up biopsies from deeper tissues in 44 cases showed presence of systemic amyloidosis in 13 cases. The sensitivity and specificity of abdominal fat pad fine needle aspiration cytology was 78% and 93%, respectively. The positive predictive value was 84% and negative predictive value 90%. CONCLUSION: Fat pad aspiration cytology is a useful screening procedure for diagnosis of systemic amyloidosis. Patients with grade 1 deposits should not undergo a toxic therapeutic regimen on the basis of fat pad cytology alone; histologic confirmation of visceral amyloid deposition in deeper tissue is advised. Patients with grades 2 and 3 deposits may undergo suitable therapy for amyloidosis.     

Molecular genetic analysis in the diagnosis of lymphoma in fine needle aspiration biopsies. II. Lymphomas versus nonlymphoid malignant tumors

The configurations of immunoglobulin genes and T-cell receptor beta chain genes were analyzed by Southern blotting in DNA derived from nonlymphoid malignant tumors and lymphomas. Gene rearrangements were not detected in any of the 35 cases of nonlymphoid malignant tumors. On the contrary, they were shown in all 14 cases of non-Hodgkin's lymphomas, 2 of 3 cases of Hodgkin's disease and 2 cases diagnosed as non-Hodgkin's lymphoma or angioimmunoblastic lymphadenopathy. The differentiation by light microscopy between lymphoma and nonlymphoid malignant tumors was a diagnostic problem in five cases; the molecular genetic analysis of DNA was contributory in all five diagnostically difficult aspirates. By gene rearrangement studies, the diagnosis of lymphoma was confirmed in two cases and nonlymphoid malignant tumors were accurately indicated in aspirates diagnosed finally as rhabdomyosarcoma (one case) and carcinoma (two cases).     

Diagnostic Alternatives Used by Veterinary Surgeons for the Recording of Udder Diseases in the Norwegian Health Card System for Cattle

A questionnaire, in which 7 cases of udder disease were described, was distributed to 890 veterinarians in Norway. They were requested to classify the cases according to the diagnostic alternatives listed in the Norwegian Health Card System for Cattle (NHCSC). The NHCSC recordings are used for progeny testing of bulls, for disease monitoring, and for research purposes. The aim of this study was to evaluate the quality of the recordings for udder diseases. The questionnaire was answered by 633 veterinary surgeons. Four cases of clinical mastitis (CM) with abnormal secretion as well as other clinical signs of inflammation were correctly classified as CM by almost 100% of the veterinary surgeons. A cow in the final stage of lactation, showing no clinical signs except for clots in the milk, was considered not to be a clinical case by more than 25% of the veterinary surgeons. A typical case of subclinical mastitis (SM) was reported as SM by 83% of the veterinary surgeons, and as CM by 16%. A subclinical case with a recent history of clots in the secretion was classified as SM by 66% of the veterinary surgeons, but almost 40% either reported CM as their sole diagnosis or considered the case to be CM in combination with SM. Of the clinical cases, those exhibiting marked local signs of inflammation and a systemic reaction were correctly classified as acute clinical mastitis (ACM) by 96%–98% of the veterinary surgeons. In the NHCSC, the diagnostic alternatives for cases of CM are ACM and chronic clinical mastitis (CCM). One case, for which the diagnosis subacute clinical mastitis was appropriate according to standard definitions, was classified as CCM by 66%), and as ACM by 6%. Based on the information given in the questionnaire, the diagnosis for 2 of the clinical cases could have been either ACM or CCM, and for both cases each of these 2 alternatives was reported by more than 43% of the veterinary surgeons. A teat lesion, which was present together with ACM in one cow, was reported by 91% of the veterinary surgeons.     

Can Creutzfeldt-Jakob disease unravel the mysteries of Alzheimer?

Recent studies on iatrogenic Creutzfeldt-Jakob disease (CJD) raised concerns that one of the hallmark lesions of Alzheimer disease (AD), amyloid-β (Aβ), may be transmitted from human-to-human. The neuropathology of AD-related lesions is complex. Therefore, many aspects need to be considered in deciding on this issue. Observations of recent studies can be summarized as follows: 1) The frequency of iatrogenic CJD cases with parencyhmal and vascular Aβ deposits is statistically higher than expected; 2) The morphology and distribution of Aβ deposition may show distinct features; 3) The pituitary and the dura mater themselves may serve as potential sources of Aβ seeds; 4) Cadaveric dura mater from 2 examined cases shows Aβ deposition; and 5) There is a lack of evidence that the clinical phenotype of AD appears following the application of cadaveric pituitary hormone or dura mater transplantation. These studies support the notion that neurodegenerative diseases have common features regarding propagation of disease-associated proteins as seeds. However, until further evidence emerges, prions of transmissible spongiform encephalopathies are the only neurodegenerative disease-related proteins proven to propagate clinicopathological phenotypes.