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.     

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.     

Early effect of poliomyelitis virus on 5'-nucleotidase activity in HeLa cells]

One of the earlier effects of poliovirus was the maintaining of a high level of Hela cells 5'-nucleotidase activity (maximal efficiency for a virus/cell ratio = 500 ID 50/cell) (ID50:50% Infections Dose). The activity decreased when cells were suspended in the presence of EDTA(EDTA:Ethylene Diamine Tetracetic Acid) and was partially restored by adding 10 mM Mg++ interactions are discussed.     

Role of calcium in the histamine release induced by D-galactosamine from rat mast cells

Rat peritoneal mast cells were isolated and purified by differential centrifugation in Ficoll. Cells pooled from three to four rats were suspended at approximately 10(6) cells/ml in a buffered salt solution and incubated for 1 h at 37 degrees C in 300 microliter volumes in the absence or presence (9 X 10(-4) M) of calcium chloride. Addition of D-galactosamine hydrochloride (DGM; 2.8 X 10(-4)M) caused (in addition to basal release) a mean +/- SEM percent histamine release of 15.7 +/- 5.2 in the presence of Ca++ and 19 +/- 4.9 in the absence of Ca++ (p greater than 0.05). It is suggested that D-galactosamine does not require extracellular Ca++ for the release of histamine from the rat mast cell.     

Net ATP synthesis by running the red cell calcium pump backwards.

Ca2+ loaded inside-out vesicles from human red blood cells, yielding C2+ into a Ca2+ free medium with 4 mM EGTA, 2 mM ADP and 10 mM phosphate, produced an excess of 14.9 pmoles . min-1 . (mg protein)-1 of ATP compared to controls in which the transmembrane Ca2+ gradient was abolished by the ionophore A 23 187.     

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.     

Intracellular topography of immunoreactive gastrin demonstrated using electron immunocytochemistry

Summary Gastrin (G)-producing cells from the mammalian gastric antrum have been investigated using computer-assisted morphometry and a novel double colloidal gold-labeled-immunoglobulin electron immunocytochemical procedure. Correlation analysis of human antral G-cells indicates (p<0.001) that a single population of granules exists with small (160 nm) electron-dense and large (240 nm) electron-lucent forms representing the extremes. Non-crossreacting region-specific antisera have been used to visualize G-17 and G-34 (progastrin) to the small electron-dense granules and G-17 to the other intermediate forms. From the results we propose a topographic segregation of immunoreactive gastrins within 2 apparently distinct granule subclasses and suggest that this may represent the pathway of granule maturation.     

Effect of oridonin, a Rabdosia diterpenoid, on radiosensitization with misonidazole

The radiosensitization brought about by oridonin, one of the Rabdosia diterpenoids, alone or in combination with misonidazole, was investigated in Chinese hamster V79 cells. The enhancement ratio of 1.92 was obtained when 0.01 mM oridonin and 1 mM misonidazole were administered to hypoxic cells under radiation. The enhancement ratios of oridonin and misonidazole for hypoxic cells were 1.16 and 1.59 respectively. Hence, a supra-additive effect was obtained by the combined treatment with these two drugs. Under aerobic conditions, no effect of 0.01 mM oridonin on the radiosensitization caused by misonidazole was observed.     

Exocytosis of secretory granules — a probable mechanism for the release of neuromodulators in invertebrate neuropiles

Summary Presynaptic terminals typically contain secretory granules, usually 100–200 nm in diameter, in addition to the smaller synaptic vesicles. Evidence is presented that granule exocytosis is a widespread phenomenon in invertebrate neuropiles. Such secretory release is apparently associated with morphologically unspecialized regions of the plasmalemma, rather than synaptic thickenings.Acknowledgments. The authors gratefully acknowledge the assistance of Mr R. M. Hewit and Mrs. S. Al-Yousuf. Supported by research grant GR/A88743 from the S.E.R.C., U.K., to D.W. Golding.     

Elektronenmikroskopische Untersuchung der phäochromen (chromaffinen) Granula in den Markzellen der Nebenniere

Summary The catechol-containing granules in the adrenal medullary cells (mouse, guinea pig, cat) are true cell organells, clearly distinguished from mitochondria; their size varies over a fairly wide range, the mean diameter being calculated at about 175 mµ in the mouse (osmium-tetroxide-fixation). They have a surrounding membrane of 100 Å thickness; their internal structure is finely granular. On the thin sections, many granules are surrounded by spaces which appear empty under the electron microscope and which are limited by double-membranes; it is most likely thatintra vitam these spaces contain lipids.

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Ausgeführt mit Unterstützung der Friedrich-Baur-Stiftung und der Deutschen Forschungsgemeinschaft.     

Degradation of pheromone biosynthesis-activating neuropeptide (PBAN) by hemolymph enzymes of the tobacco hornworm,Manduca sexta,and the corn earworm,Helicoverpa zea

The tritium-labeled bis-norleucine analog ofHelicoverpa zea pheromone biosynthesis-activating neuropeptide ([³H]NLPBAN) was incubated in vitro with hemolymph fromManduca sexta orH. zea adult females. The incubations resulted in the formation of several tritium-labeled degradation products. At a [³H]NLPBAN concentration of 0.9 μM the degradation proceeded at a very slow but physiologically plausible rate (2–10 fmol/min/μl hemolymph). The primary [³H]NLPBAN degradation reaction inM. sexta hemolymph was not inhibited by 20 μM leupeptin, 0.1 mM amastatin, 1 mM EDTA, 1 mM EGTA, 1 mM 1,10-phenanthroline, or 2 mM 4-(2-aminoethyl)benzenesulfonyl fluoride; but secondary reactions may have been affected, as some of the inhibitors changed the radio-HPLC profile of the degradation products. It is concluded that hemolymph ofM. sexta andH. zea contains peptidase(s) capable of inactivating circulating PBAN.     

Filaments of the vertebrate lens

Summary Filaments of an average diameter of 10–12 nm have been identified in the eye lens fibre cells of representative species of each vertebrate order. These filaments presumably serve a cytoskeletal role in the lens fibre cells.This study was supported by research grant Ey 0141 of the National Institutes of Health, Bethesda, Maryland, USA.     

Characterization of the (Na+-K+)-ATPase from endometrial plasma membranes of immature mammals]

The (Na+-K+)-ATPase in plasma membrane from Mammiferous endometrium is characterized by the Mg/ATP ratio equal to one, and by a distinct affinity for Na+ (1.3 mM) and K+ (2 mM). The activity is maximum for pH 7.4-7.5 in presence of Mg++ 2mM and ATP 2 mM, Na+ 140 mM and K+ 10 mM.     

壳聚糖磁性光敏剂血卟啉衍生物微球的制备及其体外表征研究

目的制备壳聚糖包覆的磁性光敏剂血卟啉衍生物微球并考察其体外表征。方法以五羰基铁为原料制备羰基铁粉纳米颗粒,非溶剂法将羰基铁粉纳米颗粒与光敏剂血卟啉衍生物HPD复合,并用壳聚糖包覆制备成壳聚糖包覆磁性血卟啉衍生物微球(CS-M-HPD)。使用电子显微镜、激光粒度仪、X-射线衍射仪和磁力测定仪考察微球的表征。结果检测结果表明该微球为球形,分散性良好,粒径大多集中在290～300 nm。微球羰基铁粉纯度高,呈现优良磁性。结论本研究实现了壳聚糖对磁性HPD的包覆,得到了单分散性磁性HPD微球,为进一步研究该微球靶向光动力治疗肿瘤的生物活性奠定了实验基础。     

Failure of calcium to stimulate Na,K-ATPase in the presence of EDTA

The effect of calcium on Na,K-ATPase activity of rat brain homogenates and its modification by the chelating agent EDTA has been investigated. In the absence of EDTA, free calcium (approximately 10(-6) mol/l) stimulates Na,K-ATPase activity; in the presence of EDTA the same concentration of free calcium is without effect on the enzyme. In the absence of EDTA the stimulation by calcium of Na,K-ATPase activity is enhanced by the additional presence of calmodulin but in the presence of EDTA, even when calmodulin is added to excess, calcium still fails to stimulate the enzyme. The possibility that EDTA interferes with an interaction between a calcium-calmodulin complex and Na,K-ATPase is discussed.     

The effect of nifedipine and verapamil on KC1-induced rhythmic contractions of guinea pig ureter in vitro

Addition of KC1 (40 mM) produced rhythmic contractions of guinea-pig ureters in vitro which were unaffected by phentolamine, atropine or tetrodotoxin. KC1 failed to elicit rhythmic contractions of ureters incubated in a Krebs solution with no added Ca++; in these conditions the addition of CaC12 in concentrations of 1.5 mM, or higher, produced rhythmic contractions whose frequency, but not amplitude, was proportional to CaC12 concentration in the bathing medium. EDTA reduced the frequency of KC1-induced rhythmic contractions without affecting their amplitude. Nifedipine and verapamil reduced both the frequency and the amplitude of KC1-induced rhythmic contraction; verapamil was more effective than nifedipine in reducing their amplitude. Urethane reduced the amplitude without significantly affecting the frequency of KC1-induced rhythmic contractions. An increase in the extracellular Ca++ concentration reverted the suppressive effect of all drugs under study. These results suggest that an influx of Ca++ from the extracellular space is responsible for the initiation of KC1-induced rhythmic contractions and is involved in the mechanism(s) which regulates their frequency, but that a separate mechanism regulates their amplitude.     

The hemolytic activity of heterocyclic N-alkyl amine oxides

Summary The N-alkyl derivatives of morpholine-, pyrrolidine-, piperidine- and perhydroasepine-N-oxides caused the rapid, temperature-dependent, hemolysis of human red blood cells. The most hemolytic were the amine oxides with alkyl groups having 14–18 carbon atoms.     

The use of liposomes in the investigation of mechanisms of asbestos damage

Asbestos-induced cell damage is initiated by a reaction at the plasma membrane. The effect of chrysotile (which is more hemolytic and releases more silicic acid than other types of asbestos) on permeability changes of liposomes has been investigated. The destabilizing effect rises when the amount of chrysotile is increased. Silicon dioxide is one major constituent which could be a hemolytic agent, and a cause of damage. It also caused an increase of permeability of liposomes.     

Responses of monkey, rabbit and dog internal carotid arteries to atrial natriuretic factor

Effects of atrial natriuretic factor (ANF) on monkey, rabbit and dog internal carotid arteries were investigated. ANF caused a concentration-dependent relaxation in arterial strips submaximally precontracted with noradrenaline, 5-hydroxytryptamine, or high-potassium solution (10-30 mM). The response was greatest in the monkey arteries and least in the dog arteries. These results suggest that there is a marked species difference in the ANF-induced relaxation of the internal carotid arteries.     

Evaluation of the ability of intact platelets to accumulate acridine orange

A new approach to the evaluation of the uptake of fluorescent probes by intact cells is described. Acridine orange (AO) was used because it can be selectively accumulated by serotonin-containing granules of platelets. Analysis of the fluorescence signal allows the estimation of the relative volume of the granules and the equilibrium coefficients for AO transport across the cytoplasm and granule membranes. The following results were obtained for human and rabbit platelets: the relative volumes of the granules were 14 +/- 1% and 29 +/- 2%, the ratios of intragranular-extracellular probe concentration were 2260 +/- 382 and 30,000 +/- 5550, and the cytoplasm-extracellular medium concentration ratios were 375 +/- 60 and 225 +/- 60, respectively.