The purification of plasma membrane from guinea pig peritoneal macrophages.
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Biomedical subjects
Publications and source records attributed to R Robineaux.
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Seven patients presenting with the characteristic clinical and laboratory findings of HCL were investigated mainly in order to find the origin of the HC with histochemical, electron microscopic and immunologic techniques. The results we obtained are somewhat contradictory, as in the recent literature, if the HC's are to the considered as being exclusively of lymphocytic or monocytic origin. The data however indicate that HCL is not of one cell type but two. One of these cells has characteristics of a lymphocyte. The other one, an ill defined cell, has mixed characteristics of both the monocyte and lymphocyte series. This hairy cell may represent a new cell line.
Observations on a 32 year old male are described. Hematological examination demonstrated leukocytosis with circulating blastosis and dystrophic hypereosinophilia of the blood and bone marrow, with cells at various stages of maturation. Cytotoxic chemotherapy led to complete remission for 5 months followed by a terminal relapse. No features in favor of an "eosinophilic collagenosis" were revealed at autopsy. Repetitive ultrastructural studies performed during evolution of the illness demonstrated considerable anomalies of the eosinophil line. The present observation thus shows the clinical, evolutional, cytological and autopsy criteria of an acute eosinophilic leukemia.
Developing eosinophils from the bone marrow of a patient with acute "eosinophilic" leukemia were characterized by electron microscopy. It was suggested that the first sequential step in granule formation occurred at the level of the endoplasmic reticulum without actual participation of the Golgi complex. Progressive densification of the former profiles, presumably mediated by Golgi vesicles, resulted in the formation of dense immature granules. Ultrastructural observations of the "leukemic" eosinophils which were generally arrested at an intermediate stage of maturation revealed also large vacuoles containing sequestered immature granules, without any indication of phagocytic activity. Morphological evidence that has been accumulated indicates that the membrane of these vacuoles fused with the cell membrane, thus being in contact with the extracellular space. These profiles strongly suggested that granules and/or granule-associated material were secreted by developing bone marrow eosinophils.
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Complement-dependent demyelinating activity of whole brain homogenate (WBH)-induced experimental allergic encephalomyelitis (EAE) sera was tested on long term tissue cultures of in vitro myelinated fetal guinea pig cerebellum. Complement-fixing (CF) auto-antibodies were shown to be the responsible agents, as demonstrated in experiments where all reagents belonged to the same species: guinea pigs of outbred (Hartley) and even of inbred (S2 or S13) strains. These antibodies were of the IgG2 class as shown by Sephadex G-200 and DEAE cellulose fractionation experiments. The corresponding auto-antigen was present in the homogenate and myelin of the central nervous system (CNS) tissue. It was different from the encephalitogenic basic protein of CNS myelin (BP), as shown in experiments where the demyelinating auto-antibodies were induced, detected, and absorbed by WBH or by CNS myelin but not by BP. They were neither induced by nor cross-reacting with cerebroside and peripheral nervous system (PNS) tissue.
We have shown in our earlier paper (4) that a ribonuclease resistant filamentous network was associated with the fibrillar structures of the nucleolus of L 929 interphase nucleus. After desoxyribonuclease digestion and oxydised DAB opacification, the nucleolar fibrillar areas became heterogenous from an electron microscopical point of view. We have concluded that an important part of these fibrillar areas are composed of DNA and that the high electron dense fibrillar structures seen after desoxyribonuclease digestion, are the primary ribosomal gene product, just transcribed on the DNA matrix.
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Electronic opacification by DAB, following ribonuclease digestion, allowed us to observe a network of DNA filaments which appeared in close relation with both intranucleolar clumps of DNA on one hand, and with the nucleolar fibrillar RNA structure on the other hand. It is inferred that the ribosomal genes may be located on these filaments of DNA.
The action of trimethylcolchicinic acid on the synthesis and excretion of proteoglycans has been studied on the L cell strain. The incorporation of precursors has been measured, and proteoglycans produced in the culture medium have been extracted and their concentration determined. The mucopolysaccharide components have been studied by electrophoresis. Control cultures produce hyaluronic acid, dermatan sulfate and very low concentrations of chondroitin 4-sulphate or 6-sulphate. Cultures treated with trimethycolchicinic acid (4 mu g/ml) produce hyaluronic acid, very high concentrations of chondroitin 4-sulphate or 6-sulphate and only traces of dermatan sulphate. So, trimethylcolchicinic acid does not modify the synthesis of hyaluronic acid: it considerably increases the production of chondroitin 4-sulphate or 6-sulphate and inhibits the production of dermatan sulphate. Protein fraction of the proteoglycans is proportionally increased in treated cultures, but there is no marked difference between amino acid concentrations of proteoglycans extracted from control and treated cultures. A slight fall in the cystine concentrations was the only change in the amino acid content of proteoglycans extracted from treated cultures. A hypothesis to explain these results is discussed.