Acute alcoholic myopathy. Enzyme histochemistry and electron microscopic findings

Five cases of acute alcoholic myopathy were studied by light microscopy and histochemical techniques. Three cases were studied by electron microscopy. The findings were uniform showing rhabdomyolysis and chronic interstitial inflammation. Type I fibers were predominantly affected with patchy loss of oxidative enzyme activity in some fibers.The electron microscopic observations disclosed conspicuous mitochondrial abnormalities, characterized by partial or total dissolution of mitochondria with disorganization and swelling of the cristae. Bizarre paracrystalline rectangular inclusions as well as amorphous osmiophilic inclusions were noted in some degenerating mitochondria. It was concluded that such enzyme histochemical and mitochondrial changes form a non-specific response to a variety of agents injurious to skeletal muscle.     

Enzyme histochemistry on skeletal muscle of the human foetus.

The differentiation of fibre types in developing human skeletal muscle was studied. The material consisted of muscle samples from different muscles of 86 foetuses (abortions) between 12 weeks gestation and delivery and 50 children 1 day to 7 years old. The latter samples were obtained at surgery. Histochemical stains for myofibrillar ATPase were made after preincubations at pH 4.3, 4.6 and 10.3 in order to identify the subgroups A and B of type II fibres and undifferentiated fibres (type II C). Stains for glycogen and lipids were also performed as well as for NADH-diaphorase and alpha-glycerophosphate dehydrogenase. After 20 weeks gestation a few large size type I fibers could be found in some muscles, but not until after the 30th week were some type II A fibres seen. During the last 3 months of gestation a very rapid further differentiation occurred, but at delivery the differentiation process was still not completed. At birth 15-20% of the fibres were classified as undifferentiated. This picture only gradually changed with a slow increase in the number of type I, II A and II B fibres. The stains for metabolic enzymes and substrates were pale until late in foetal life when some distinction between fibre types became discernible.     

Characterization of gap junctions between cultured leptomeningeal cells.

Leptomeningeal cells in intact meninges or dissociated and cultured for 2 h to several weeks were dye-coupled (Lucifer yellow), and voltage-clamped pairs of freshly dissociated leptomeningeal cells were well coupled electrically. Unitary conductances of junctional channels were predominantly 40-90 pS. Junctional conductance was reversibly reduced by 2 mM halothane, 1 mM heptanol and 100% CO2 and was increased by 1 mM 8 Br-cAMP. Two gap junction proteins, connexin 26 and connexin 43, were identified between leptomeningeal cells using immunocytochemical methods; Northern blot analyses of RNA isolated from cultured leptomeningeal cells showed specific hybridization to cDNAs encoding connexins 26 and 43, but not to a cDNA encoding connexin 32. These studies demonstrate co-expression of two connexins in a single cell type in the nervous system; biophysical properties do not differ significantly from those of astrocytes and cardiac myocytes, which express only connexin 43.     

Implantation of cultured human leptomeningeal cells into rat brain

Since previous studies have shown that cells cultured from human leptomeninges can express neuronal and glial antigens under appropriate culture conditions [DeGiorgio L. A. et al. (1994) J. Neurol. Sci.124, 141–148; Bernstein J. J. et al. (1996) Int. J. Devl Neurosci.14(5), 681–687], we have studied the developmental characteristics of these cells further by grafting them into young adult rat brains. Cells were labeled in culture with Fast Blue and were identified unequivocally by hybridization with nick-translated human DNA. Intensely Fast Blue positive human leptomeningeal cells were concentrated in the implant pocket and adjacent rat leptomeninges at one and two weeks postimplant. Human and rat leptomeningeal cells were similar morphologically and were equally immunopositive for vimentin and fibronectin. Implanted human cells did not express the neuronal and glial proteins they had in vitro. Cells which hybridized with human DNA corresponded to the intensely Fast Blue positive cells. Small groups of human DNA hybridizing cells were also observed in the choroid plexus. Less intensely Fast Blue positive neurons and glia were found in the brain, but these hybridized with rat DNA. A minority of human leptomeningeal cells implanted into rat brain are subsequently found in host leptomeninges where they demonstrate properties characteristic of leptomeningeal fibroblasts. Small numbers of implanted cells can survive for two weeks.     

Numeric assessment of leptomeningeal nerve cells. A semiquantitative study

Since 1900, several authors have described the presence of mature neurons in the leptominges. Using human material, a semiquantitative evaluation of their number has been performed. The distribution pattern of these nerve cell populations is presented. The role of the leptomeningeal neurons in the regulation of the cerebral microcirculation is discussed.     

Acid phosphatase activity of cerebrospinal fluid cells in leptomeningeal haemorrhage

Summary One hundred and twenty samples of haemorrhagic spinal fluid were examined by acid phosphatase staining. This enzyme activity starts to appear in mono-histiocytic cells 2 days after bleeding and increases up to the 5th day. After 1 week the activity decreases rapidly. Similar results are found in mixtures of incubated clear spinal fluid, to which blood is added. Acid phosphatase staining is a useful additional method for determination of the age of a leptomeningeal bleed.     

Expression of a membrane-bound form of the ferroxidase ceruloplasmin by leptomeningeal cells

Ceruloplasmin is a key enzyme involved in detoxifying ferrous iron, which can generate free radicals. The secreted form of ceruloplasmin is produced by the liver and is abundant in serum. We have previously identified a membrane-bound glycosylphosphatidylinositol (GPI)-anchored form of ceruloplasmin (GPI-Cp) that is expressed by astrocytes in the central nervous system (CNS) (Patel and David. 1997. J Biol Chem 272:20185-20190). We now provide direct evidence that rat leptomeningeal cells, which cover the surface of the brain, also express GPI-Cp. The expression of GPI-Cp on the surface of these cells increases with postnatal development and is regulated in vitro by cell density, time in culture, and various extracellular matrix molecules. The expression of GPI-Cp also appears to be regulated differently in astrocytes and leptomeningeal cells in vitro. The abundant expression of GPI-Cp on the surface of leptomeningeal cells suggests that these cells play a role in antioxidant defense along the surface of the postnatal CNS possibly by detoxifying the cerebrospinal fluid.     

Some remarks on the morphology and histochemistry of the so-called Opalski cells

Summary The morphology of Opalski cells and histochemical properties of substances filling their cytoplasm are discussed. In the morphological picture of Opalski cells special attention has been paid to the presence of residual cell processes which are visible both in the routin cellular stainings and in gold-sublimate Cajal impregnation. Their presence and positive metalic impregnation in the author's opinion, are conclusive for astrocytic origin of Opalski cells. The second fundamental morphological feature of Opalski cells consisted in the presence of PAS-positive granulations filling their cytoplasm. On the basis of the histochemical tests performed, the author considered these granular substances as acid mucopolysaccharides bound with cellular proteins containing copper. Opalski cells, being a degenerative forms of astroglia appear as the result of disturbances in the intracellular metabolism of astrocytes. Subsequently to this metabolic disorders in their cytoplasm, abnormal metabolites of the above described character are accumulated. Opalski cells originate from Alzheimer cells type I, however, it seems, that their direct development from typical astrocytes avoiding the stage of Alzheimer cells, is also possible. A few examples of Opalski cells could be seen also apart from hepato-lenticular degeneration, for instance in cases hepatogenic encephalopathy.

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Zusammenfassung Die Morphologie der Opalski-Zellen und die histochemischen Eigenschaften der ihr Cytoplasma erfüllenden Stoffe werden dargelegt. Im morphologischen Bild der Opalski-Zellen wird besonders das Vorhandensein von Zellfortsatzresten hervorgehoben, die sowohl in den Routinezellfärbungen als auch in der Cajal-Goldsublimat-Imprägnation zu sehen sind, was nach der Meinung des Autors auf die Herkunft der Opalski-Zellen aus Astrocyten hinweist. Die zweite morphologische Haupteigenschaft der Opalski-Zellen machen die das Cytoplasma erfüllenden PAS-positive Granula aus. Auf Grund der durchgeführten histochemischen Tests werden diese granulären Substanzen als saure Mucopolysaccharide in Bindung an kupferhaltige Zellproteine betrachtet. Die Opalski-Zellen als degenerative Astrogliaformen erscheinen als Ergebnis von Störungen im intracellulären Stoffwechsel der Astrocyten; dieser zufolge werden abnorme Metaboliten der beschriebenen Art im Cytoplasma angehäuft. Die Opalski-Zellen leiten sich aus Zellen vom Alzheimertyp I ab; es erscheint jedoch möglich, daß sie sich auch direkt aus typischen Astrocyten entwickeln. Vereinzelte Opalski-Zellen kommen auch außerhalb der hepatolenticulären Degeneration, z.B. in Fällen von hepatogener Encephalopathie vor.

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With 8 Figures in the Text     

Diagnosis of leptomeningeal metastasis

Forty-one cases of leptomeningeal metastases from solid and hematological tumours were studied clinically and pathologically. Neurological symptoms and signs, cerebrospinal fluid characteristics and radiographic appearance were reviewed. The role of the biochemical markers, computer tomography scans of the brain and myelography in the diagnosis of patients with leptomeningeal metastasis were determined. The subject of the study was an investigation of the value of various diagnostic procedures to increase the accuracy of the diagnosis leptomeningeal metastasis.     

Gap junction disappearance in astrocytes and leptomeningeal cells as a consequence of protozoan infection

Trypanosoma cruzi and Toxoplasma gondii are protozoan parasites capable of causing infections of the nervous system. In order to determine effects of infection by these organisms on intercellular communication in the brain, dye coupling and connexin abundance and distribution were examined in leptomeningeal cells and astrocytes infected with T. cruzi or T. gondii. For both cell types infected with either type of protozoan parasite, intercellular diffusion of intracellularly injected Lucifer Yellow was dramatically reduced. Immunocytochemistry with antibodies specific for connexin43 (in astrocytes) or both connexin43 and connexin26 (for leptomeningeal cells) demonstrated that punctate gap junctional staining was much reduced in infected cells, although uninfected neighbors could display normal connexin abundance and distribution. Western blot analyses revealed that connexin43 abundance in both cell types infected with either parasite was similar to that in uninfected cells. Phosphorylation state of connexin43 (inferred from electrophoretic mobility of connexin43 isoforms) was not significantly affected by the infection process. Immunocytochemistry of whole brains from animals acutely infected with either parasite also showed a marked reduction in connexin43 expression. We conclude that infection of both types of brain cells with either protozoan parasite results in a loss of intercellular communication and organized gap junction plaques without affecting expression levels or posttranslational processing of gap junction proteins. Presumably, these changes in gap junction distribution result from altered targeting of the junctional protein to the plasma membrane, and/or from changes in assembly of subunits into functional channels.     

The amyloid beta-protein precursor is localized in smooth muscle cells of leptomeningeal vessels

We examined leptomeningeal vessels using 2 different antisera which recognized both terminal ends of amyloid beta-protein precursor (APP). Both antisera recognized a 120-kDa protein from leptomeningeal vessels. Immunocytochemistry showed that APP was localized in the smooth muscle cells of leptomeningeal vessels. These results indicated that the smooth muscle cells of leptomeningeal vessels contained full-length native APP, and that the cerebrovascular amyloid in the leptomeningeal vessels was derived from this APP.     

Post-traumatic intradiploic leptomeningeal fistula and cyst.

A 59 year old female patient presented with ataxia and difficulty in walking. The neurological examination revealed right homonymous hemianopia and ataxia. Radiographic evaluation revealed a large occipital intradiploic cyst mainly in the left suboccipital area. There was also moderate hydrocephalus and encephalomalacia of the left occipital pole. Bone window studies also demonstrated a growing fracture extending from the upper pole of the cyst to the vertex. Both pathologies were attributed to child abuse the patient suffered when she was a child. At first surgery, decompression of the cerebellum was followed by duroplasty and acrylic cranioplasty to the posterior cranial fossa. A month later, a shunt had to be inserted for hydrocephalus. At 7 months postoperatively, the patient is well and free of any symptoms or recurrence.     

Rhabdoid and papillary meningioma with leptomeningeal dissemination

Rhabdoid meningioma is a rare variant of meningioma classified as grade III under the new World Health Organization (WHO) classification of brain tumors. Although this tumor is known for its aggressive behavior, dissemination into cerebral spinal fluid (CSF) is extremely rare. We report here a case of rhabdoid meningioma in a young man, operated on twice previously, who presented with multiple CSF areas of seeding in the brain and spinal cord. The imaging findings for this tumor, including diffusion and perfusion MR sequences, are highlighted. This particular histological subtype of meningioma has a poor prognosis and must be treated aggressively.