Identification of hemolytic granules isolated from human myocardial cells

Human myocardial cells from fresh autopsy material contained granules which possessed hemolytic activity against guinea pig and rabbit erythrocytes. The hemolytic granules, which had a density of 1.02 and a diameter of 200-300 nm, were recovered as a microsome fraction from subcellular homogenates of human myocardial cells by differential centrifugation in 300 mM sucrose containing 0.1 mM PMSF and 10 mM EDTA. The membrane lesions caused by the granules were ring-like structures with an internal diameter of about 10-17 nm, analogous to that caused by perforin- and complement-induced lysis. However, the requirement for divalent cation differed from that for perforin-induced lysis, since the microsome-mediated lysis occurred in the presence of EDTA.     

Identification of hemolytic granules isolated from human myocardial cells

Summary Human myocardial cells from fresh autopsy material contained granules which possessed hemolytic activity against guinea pig and rabbit erythrocytes. The hemolytic granules, which had a density of 1.02 and a diameter of 200–300 nm, were recovered as a microsome fraction from subcellular homogenates of human myocardial cells by differential centrifugation in 300 mM sucrose containing 0.1 mM PMSF and 10 mM EDTA. The membrane lesions caused by the granules were ring-like structures with an internal diameter of about 10–17 nm, analogous to that caused by perforin- and complement-induced lysis. However, the requirement for divalent cation differed from that for perforin-induced lysis, since the microsome-mediated lysis occurred in the presence of EDTA.     

Ultrastructure of neurons of nucleus preopticus and pars lateralis of the nucleus lateralis tuberis in Gambusia (Teleosta poeciliidae)]

The nucleus preopticus (NPO) and the pars lateralis of the nucleus lateralis tuberis (NLT) have been examined at the ultrastructural level. In the NPO two types of large neurons occur, both containing 140-150 nm secretory granules. The pars lateralis of the NLT contains only one cell type with similar granules. Both of these nuclei may be the source of type A fibres innervating the gonadotropic cells of Gambusia.     

Claudin-17 forms tight junction channels with distinct anion selectivity

Barrier properties of tight junctions are determined by the claudin protein family. Many claudins seal this barrier, but others form paracellular channels. Among these, no claudins with general and clear-cut anion selectivity have yet been described, while for claudin-10a and claudin-4, only circumstantial or small anion selectivities have been shown. A claudin with unknown function and tissue distribution is claudin-17. We characterized claudin-17 by overexpression and knock-down in two renal cell lines. Overexpression in MDCK C7 cell layers caused a threefold increase in paracellular anion permeability and switched these cells from cation- to anion-selective. Knockdown in LLC-PK(1) cells indorsed the finding of claudin-17-based anion channels. Mutagenesis revealed that claudin-17 anion selectivity critically depends on a positive charge at position 65. Claudin-17 expression was found in two organs: marginal in brain but abundant in kidney, where expression was intense in proximal tubules and gradually decreased towards distal segments. As claudin-17 is predominantly expressed in proximal nephrons, which exhibit substantial, though molecularly not defined, paracellular chloride reabsorption, we suggest that claudin-17 has a unique physiological function in this process. In conclusion, claudin-17 forms channels within tight junctions with distinct anion preference.     

The subcommissural organ of the lizardLacerta s. sicula raf. ultrastructure during the winter

Summary The secretory activity of the SCO cells ofLacerta s.sicula Raf. is strongly reduced during the winter. Such reduction is documented by the decrease of the number of secretory granules type A and B described in previous papers in the summer SCO cells. Also the sacks of RER filled with electron-dense material (type C secretion) are very few; in their place there are, in the basal region of the cells, large vacuoles. In the distal region of the cells, at the free cell surface, a pronounced increase in the number of microvilli is noticed.     

Giant granules in adrenaline-secreting chromaffin cells of lizard adrenal glands after metyrapone administration

Summary Secretory granules of extraordinary size, some of them bigger than the cell nucleus, abound in the adrenaline cells of lizard adrenals after metyrapone injections during 7 days. In these granules, the bounding membrane is studded with ribosomes, and the core is formed by rounded small subunits. Some granules of this type are also found in noradrenaline cells. They may represent an exceptionally increased elaboration and storage of adrenaline, induced by metyrapone probably through its action on steroidogenic tissue.This research forms part of project No. 31.26.S1-1154 supported by the Consejo Nacional de Investigaciones Científícas y Tecnológicas.     

Ultrastructure of gonadotropic cells of the normal eel and after estradiol injection]

Male and female silver eels, in fresh water or sea water, have few gonadotrophs (GTH) poorly differentiated. An oestradiol (E2) treatment induces their development: a well developed Golgi area, abundant secretory granules (200-500 nm), dilated cisterna and large globules 1.2-2.2 micron) are observed. A positive feedback may control gonadotropin synthesis in immature eels. No macroscopic effect is discernible on the gonad.     

The multifaceted Paneth cell

Paneth cells (PCs) were described over a century ago as granulated cells located at the base of small intestinal crypts, the 'crypts of Lieberkühn.' Various histochemical staining procedures were developed that identified PCs based on their distinctive granule staining pattern. Early on, PCs were proposed to perform a specialized function other than absorption of digested nutrients, the predominant task of the small intestinal epithelium. Since then, many constituents of the PC granules have been biochemically characterized. The presence of various granule-associated antimicrobial substances and their release upon microbial challenge suggest that PCs function as specialized defense cells in the small intestine. Altered resistance to microbial infection in animal models with disrupted or augmented PC function provides further support for the host defense role of PCs. Other PC components suggest that PCs may also participate in the regulation of lumenal ionic composition, crypt development, digestion, and intestinal inflammation. Received 6 June 2001; received after revision 26 July 2001; accepted 27 July 2001     

Peroxidase activity of neutrophil granules in two cases of congenital myeloperoxidase deficiency]

The neutrophils and monocytes of two patients with hereditary myeloperoxidase deficiency lacked myeloperoxidase activity as determined by light and electron microscopic cytochemical staining. Using Graham-Karnovsky media, azurophils of neutrophils were devoid of peroxidase whereas all eosinophilic and basophilic granules exhibited normal peroxidase activity. After incubation in alkaline diaminobenzidine media which stains the catalase of microperoxisomes, some small granules were seen to be strongly stained in both immature and mature neutrophils. These small granules were distinct from all other neutrophilic granules which lacked a positive reaction. Only, in the presence of cyanide or aminotriazole, peroxidatic activity was also detected in some ellipsoid azurophils. This observation suggests that these substances activated an oxidase whose nature is discussed.     

Are the pores of intramembrane particles of postsynaptic membrane transmitter-dependent channels?

The rat neonatal cerebellar cortex has been studied using the freeze fracture technique. In the dendritic postsynaptic membrane, intramembrane particles containing an electron-dense central area have been detected. This type of area could be a platinum aggregate within a channel which, crossing the particle, may connect the postsynaptic cytoplasm with the synaptic cleft.     

Purification of the cytosol oestradiol-receptor complex from foetal guinea-pig uterus using electrofocusing on polyacrylamide plates

The 3H-oestradiol receptor complex obtained from the cytosol fraction of foetal guinea-pig uterus was purified by the following steps: column chromatography in Sephadex G-15 and Ultrogel, and by electrofocusing on polyacrylamide plates. In the final step a concentration of 15-17% of the foetal uterine oestradiol receptor protein was obtained. The isoelectric point (pl) of this receptor was determined to be 6.1-6.2.     

Intranukleäre virusähnliche Partikeln in einem Mammakarzinom

Summary By electron microscopic examination of a solid, chiefly scirrhous mammary cancer, 3 kinds of unusual nuclear inclusions were found: (a) Electron dense particles of a 54–80 nm diameter, whose outlines appear mainly hexagonal, which points at an icosahedral structure, (b) Clusters of granules with a diameter of 200–300å, respectively 300–400å, at whose circumference the larger particles appear, (c) Bundles of filaments in close association and continuous with the granules. In all 3 nuclear inclusions there are subunits of 40–45å. The comparison between these results and experiments published suggest that these nuclear inclusions are (a) virus particles, (b) virus at an early stage of development and (c) virus protein subunits.     

Further studies on gel filtration of urinary substances.

Urine specimens from healthy children were concentrated and filtered through Sephadex G-100 and G-25. The eluate was recovered in fractions of 5 ml each. Total hexoses, galactose, total proteins and total uronic acids were determined on each fraction. Total hexoses and galactose were eluted in 6 distinct peaks. The uronic acid containing GAG were eluted in 4 small peaks on G-25 and 1 peak on G-100. This peak is eluted always with high amounts of galactose containing glycoproteins.     

Types of nerve terminals in fetal and neonatal rabbit myocardium

With the use of electron microscopy 4 types of axonal profiles were observed in the developing myocardium of rabbits: 1) adrenergic axons which contained mainly small dense-core vesicles and which presumably can store 5-hydroxy-dopamine; 2) cholinergic axons which contained small clear synaptic vesicles and which were acetylcholinesterase-positive; 3) axons which contained large vesicles filled with moderately electron-dense material and which resembled purinergic axons; and 4) profiles filled with mitochondria, vesicles of various sizes, lysosome-like bodies, and microtubules and which resembled sensory terminals.     

Further studies on gel filtration of urinary substances

Summary Urine specimens from healthy children were concentrated and filtered through Sephadex G-100 and G-25. The eluate was recovered in fractions of 5 ml each. Total hexoses, galactose, total proteins and total uronic acids were determined on each fraction. Total hexoses and galactose were eluted in 6 distinct peaks. The uronic acid containing GAG were eluted in 4 small peaks on G-25 and 1 peak on G-100. This peak is eluted always with high amounts of galactose containing glycoproteins.This work was partially supported by research grants from the CONICET, República Argentina.     

The EM immunocytochemical demonstration of lysozyme in macrophage giant cells in sarcoidosis.

The giant cells (multinucleate macrophages) of human sarcoidosis have been shown by the unlabelled antibody immunoperoxidase technique at electron microscope level to contain lysozyme within cytoplasmic granules.     

Partial purification and characterization of acid phosphatase from sporulated oocysts of Eimeria tenella

Acid phosphatase of Eimeria tenella oocysts (Peak II) was purified 77-fold with a recovery of 26% using protamine sulfate precipitation, DEAE-cellulose chromatography and Sephadex G-200 gel filtration. This enzyme occurs in multiple forms as indicated by two peaks which can be separated by DEAE-cellulose chromatography and polyacrylamide gel electrophoresis. The partially purified enzyme has optimal activity at pH 4.5. With p-nitrophenyl phosphate the Km and Vmax values for (Peak II) were 25 mM and 1.57 mumol/min/mg protein, respectively. The enzyme (Peak II) is strongly inhibited by Hg++, Cu++, iodoacetamide, fluoride and molybdate. Tartrate and other divalent metal ions have no effect on enzyme activity. The partially purified Peak II phosphatase is not a glycoprotein as it is not absorbed on concanavalin-A Sepharose and its treatment with bacterial neuraminidase does not alter its elution profile through DEAE cellulose.     

Melanogenic melanocytes in human sebaceous glands.

Electron microscopic observations revealed for the first time a small number of active melanocytes synthesizing distinctive melanin-containing organelles (melanin granules)in the ducts and acini of human sebaceous glands.     

Melanogenic melanocytes in human sebaceous glands

Summary Electron microscopic observations revealed for the first time a small number of active melanocytes synthesizing distinctive melanin-containing organelles (melanin granules) in the ducts and acini of human sebaceous glands.     

Agglutination of erythrocytes using lectin-labeled spacers

Summary The mechanism of agglutination of human erythrocytes by lectin, studied using lectin-labeled gold granules as spacers (5–64 nm in size), was found to depend upon the size of the spacer but not upon the number and the mobility of the receptors.