Analysis of nuclear sugar-binding components in undifferentiated and in vitro differentiated human promyelocytic leukemia cells (HL60)

The nuclear sugar-binding components (i.e., lectinlike molecules) were analyzed using isolated and membrane-depleted nuclei after incubation in the presence of fluorescein-labeled neoglycoproteins. This analysis was performed before and during the in vitro differentiation of HL60 cells into monocytes by PMA treatment and into granulocytes by DMSO treatment. The nucleoli of undifferentiated and differentiated HL60 cells were not labeled, unlike the nucleoli of other mammalian cells studied so far. This peculiarity allowed us to quantitatively analyze by flow cytometry the changes in the lectin activity associated with the extranucleolar territories enriched in ribonucleoprotein complexes. The neoglycoprotein binding was found to be significantly lower in differentiated than in undifferentiated cells. The decrease in neoglycoprotein binding was observed within the first 24 h of DMSO or PMA treatment, just before the arrest of DNA synthesis. Taking into account that the granulocytic differentiation required 72 h of chemical treatment, the extra-nucleolar lectins might be involved in modulation of the DNA synthesis rather than in phenotypic differentiation. These data are discussed in an attempt to reconcile the association of lectins with RNP complexes and their possible involvement in modulation of HL60 cell proliferation.     

Antitumor effect of heat-killed Aspergillus fumigatus mycelium in a mouse model

Summary A suspension of heat-killed Aspergillus fumigatus mycelium inhibited the growth of a chemically-induced mouse bladder tumor (MBT). Tumor growth was inhibited when the mycelium was injected into mice in a mixture with the tumor cells, when injected into growing tumors, and when introduced IP at the time tumor cells were injected into the hind leg muscle. In the concentrations that affected tumor growth no toxicity of the fungus preparation was observed. The fungal suspension was more effective against MBT than a Corynebacterium parvum strain known to be a potent biologic response modifier. A significant increase in the number of mouse peritoneal exudate cells (PEC) was noted following inoculation with the mycelium. The induced PEC were cytotoxic to the tumor cells in vivo, suggesting that at least part of the tumor inhibition by the mycelium is host-mediated.     

Diphenoloxidases in various forms of myopathy which are transmitted by different genetic mechanisms

Summary In a previous article (Demos et al. 1981), we reported a significant and specific reduction of the activity index (AI) of the diphenoloxidases (DPox) in patients and heterozygotes with progressive Duchenne muscular dystrophy (DMD), which is transmitted genetically by female subjects by a sex-linked recessive mechanism (SLR). This same anomaly was detected in patients suffering from other types of dystrophy: Becker, limbgirdle, fascio-scapulo-humeral, and in heterozygotes, of either sex in diseases transmitted by an obviously recessive autosomic mechanism. These anomalies were detected using blood spots collected on absorbent paper and stored at 4°C for differnt periods. They were of the same type as had previously been detected using blood platelets (Demos 1973).     

Assay of tannase (tanin acylhydrolase EC 3.1.1.20) by gas chromatography

A gas chromatographic method has been satisfactorily developed for the determination of gallic acid after enzymatic hydrolysis of methyl gallate by fungal tannase. The technique described herein permits a specific, quantitative analysis of the enzyme. The separate determination of both soluble and fixed tannase is described.     

Reverse-phase high-pressure liquid chromatography of uridine diphosphate N-acetylmuramyl peptide precursors of bacterial cell wall peptidoglycan

A series of seven different UDP-N-acetylmuramyl peptide precursors of bacterial cell wall peptidoglycan was examined by reverse-phase high-pressure liquid chromatography. Mixtures of these compounds were successfully and rapidly analyzed by using a Waters μBondapak C₁₈ column as a stationary phase and isocratic elutions with 0.05 m ammonium phosphate or formate buffers of appopriate pH. Furthermore, their accurate quantitation could also be readily achieved by reverse-phase high-pressure liquid chromatography. All these techniques should be extremely useful for the purification of these compounds and for a wide range of biochemical studies concerning the cytoplasmic steps of the biosynthesis of peptidoglycan.     

Conformational study of native and acid-dissociated states of porcine lutropin (pLH) by vacuum circular dichroism and ultraviolet absorption spectroscopy

Porcine lutropin shares with ovine lutropin common structural features. They exhibit identical vacuum circular dichroism down to 170 nm with characteristic negative bands at 173, 194, and 210 nm. The band at 210 ± 1 nm is shifted to 201 nm upon dissociation with disappearance of the 194-nm band. For the two hormones the acid transition involves a significant loss of the three recognized periodic structures α helix, β sheet, and β turns of type II, unshields near the same number of tyrosyl residues (2.2 ± 0.7), and gives rise to an identical absorption difference at 287 nm in a two-step mechanism. However, pLH also exhibits noticeable differences: 5 to 10 times lower rates of acid transition with a lower pK_a (3.7 ± 0.1) and different transition behavior of tyrosine residues compared to ovine lutropin.     

Evidence for Glucocorticoid Target Cells in the Rat Optic Nerve. Hormone Binding and Glycerolphosphate Dehydrogenase Induction

Abstract: Biochemical evidence suggests that neuroglia are responsive to glucocorticoids, yet previous studies of glucocorticoid localization have typically failed to demonstrate significant uptake by neuroglial cells. To further investigate this problem, we measured glycerol-3-phosphate dehydrogenase (GPDH) activity and glucocorticoid receptor binding capacity in normal rat optic nerves and in those undergoing Wallerian (axonal) degeneration. Binding studies were also performed on hippocampus and anterior pituitary for comparison purposes. Normal optic nerve preparations possessed a high level of GPDH activity that was glucocorticoid-inducible and that increased further following axonal degeneration. Antibody inactivation experiments demonstrated the presence of more enzyme molecules in the degenerating nerve preparations. Correlative immunocytochemical studies found GPDH-positive reaction product only in morphologically identified oligodendrocytes, a result that is consistent with the previously reported localization of this enzyme in rat brain. Optic nerve cytosol fractions displayed substantial high-affinity binding of both dexamethasone (DEX) and corticosterone (CORT) that, like GPDH, was elevated approximately twofold in degenerating nerves. Finally, in vivo accumulation of [³H]DEX and [³H]CORT by optic nerve and other myelinated tracts was examined using nuclear isolation and autoradiographic methods. Although neither steroid was found to be heavily concentrated by these tissues in vivo , a small preference for DEX was observed in the nuclear uptake experiments. These results are discussed in terms of the hypothesis that glial cells are targets for glucocorticoid hormones.     

The Quaternary Structure of Chicken Acetylcholinesterase and Butyrylcholinesterase; Effect of Collagenase and Trypsin

Acetylcholinesterase (EC 3.1.1.7.; AChE) and butyrylcholinesterase (EC 3.1.1.8.; BuChE) from chicken muscle exist as sets of structurally homologous forms with very similar properties. The collagenase sensitivity and aggregation properties of the 'heavy' forms of both enzymes indicate that they possess a collagen-like tail, and their stepwise dissociation by trypsin confirms that they correspond to triple (A12) and double (A8) collagen-tailed tetramers. In addition to this dissociating effect, trypsin digests an important fraction of the catalytic units of AChE, in a progressive manner, removing as much as 30% of the enzyme's mass, without inactivation of the tetramers and of the tailed molecules. The trypsin-modified AChE forms closely resemble the corresponding mammalian AChE forms in their hydrodynamic properties. It is not known whether the trypsin-digestible peptides, which do not appear to be involved in the ionic or hydrophobic interactions of the enzymes, are a fragment of the catalytic subunit or whether they constitute distinct polypeptides.     

Evidence for sedative effects of low doses of morphine in mice involving receptors insensitive to naloxone

Low doses of morphine (0.30–2.5 mg/kg) decrease in a dose-dependent manner spontaneous climbing behaviour in mice. This effect is not modified by administration of naloxone at doses up to 1.25 mg/kg. These morphine doses do not modify the locomotor activity but, when they are associated with naloxone (0.5 mg/kg), an obvious inhibition occurs. In rats, a hyperactivity follows the akinesia produced by a morphine administration (10 mg/kg). This hyperactivity is changed into a significant hypokinesia when the animals are treated with naloxone (0.05 mg/kg). These results might reveal a dual effect of low doses of morphine, the excitatory effect of morphine being antagonized by naloxone whereas no action on the sedative effect is observed.