On the binding of steroid sulfates to albumin

Summary ³H-Labeled steroid sulfates, sulfate of estrone (E₁S) or dehydroepiandrosterone (DHAS), were dialyzed against delipidated human serum albumin or human plasma in the presence of increasing amounts of competing non-labeled sulfates (DHAS or E₁S). The apparent equilibrium constants (K) of the tracers did not measuraby change at concentrations of the non-radioactive sulfates below 10^–5 mol/l. At higher concentrations, K decreased gradually. The apparent equilibrium constant of³H-E₁S was diminished by plasma in a similar fashion. It may be concluded that albumin possesses one strong, non-specific binding site. This site, however, does not seem to be utilized for the binding of E₁S in vivo, because of its preferetial occupation by other ligands. This may be true for other steroid sulfates as well, depending on their relative abundance in plasma.Acknowledgment. This investigation received financial support from the Special Programme of Research, Development and Research Training in Human Reproduction, World Health Organisation, and from the Ford Foundation.     

Testosterone secretion by Mongolian gerbil interstitial cells during short-term incubation depends on androgen precursors and serum proteins but not on gonadotrophins

Interstitial cells from the testes of the Mongolian gerbil have been used to investigate the effects of serum proteins on testosterone production stimulated by hCG and steroidal precursors. Short-term incubation of interstitial cells with progesterone or DHEA resulted in a rapid increase of testosterone secretion; this effect was even more pronounced in the presence of calf serum. On the other hand, addition of hCG (10 mIU) had no significant effect on testosterone release during the 30-min incubation. These results demonstrate that the magnitude of the steroidogenic response of short-term incubated interstitial cells is a complex function, mainly of precursor concentrations and binding capacities of serum proteins but not of gonadotrophins.     

Testosterone secretion by Mongolian gerbil interstitial cells during short-term incubation depends on androgen precursors and serum proteins but not on gonadotrophins

Summary Interstitial cells from the testes of the Mongolian gerbil have been used to investigate the effects of serum proteins on testosterone production stimulated by hCG and steroidal precursors. Short-term incubation of interstitial cells with progesterone or DHEA resulted in a rapid increase of testosterone secretion; this effect was even more pronounced in the presence of calf serum. On the other hand, addition of hCG (10 mIU) had no significant effect on testosterone release during the 30-min incubation. These results demonstrate that the magnitude of the steroidogenic response of short-term incubated interstitial cells is a complex function, mainly of precursor concentrations and binding capacities of serum proteins but not of gonadotrophins.8 October 1986     

On the distribution of plasma L-asparaginase

Summary The guinea-pig,Cavia porcellus, is unusual in possessing plasma L-asparaginase, an enzyme with anti-tumor activity. 21 additional species have been examined as to the presence of this enzyme: the results confirm and extend its remarkably limited species distribution.This work was supported in part by grant GM/CA 25378 from the National Institute of General Medical Sciences, National Institutes of Health.We are grateful to dr Hanna Hensch, Philadelphia Zoological Garden, and Ms Joan Whitla and Dr Richard J. Montali, National Zoological Park/Smithsonian Institution, for samples of frozen serum.To whom correspondence should be addressed.     

Preferential excretion of glycated albumin in C57BL-Ks-J mice: Effects of diabetes

Urinary excretion of glycated albumin was quantitated in genetically hyperglycemic mice (C57BL-Ks-J, db/db mice), a model for non-insulin-dependent diabetes mellitus, and compared with their non-diabetic littermates. The data indicated a preferential excretion of glycated albumin in non-diabetic mice. This phenomenon of editing of glycated albumin is decreased significantly in diabetic mice. Quantitative measurements of overall excretion of glycated albumin suggested that the loss of editing in diabetic mice is due to the dilution of glycated albumin by the unmodified albumin which is excreted in large amounts in diabetic mice. Therefore, the loss of editing observed in this model resembled the one we characterized in insulin-dependent diabetic humans and a streptozotocin-diabetic rat model³.     

α1-Adrenergic stimulation of ketogenesis and fatty acid oxidation is associated with inhibition of lipogenesis in rat hepatocytes

Summary The effect of norepinephrine on fatty acid synthesis (³H₂O incorporation into fatty acids), on fatty acid oxidation to CO₂ and on ketogenesis was studied in isolated hepatocytes of fed rats. After incubation with norepinephrine (50 M), lipogenesis was lower (5.7±1.1 nmoles³H₂O incorporated into fatty acids/mg dry weight/30 min) than in controls (7.5±1.7; n=6, p<0.02). In contrast, (1-¹⁴C) palmitate conversion into total ketone bodies was increased to 10.9±1.8 nmoles/mg/30 min with norepinephrine, vs 8.5±1.6 in controls (p<0.05), and more (1-¹⁴C) palmitate was converted to¹⁴CO₂ with norepinephrine than in controls (1.48±0.10 nmoles/mg/30 min vs 1.06±0.11, p<0.05). The inhibitory effect of norepinephrine on lipogenesis was abolished by addition of the ₁-receptor blocker prazosin, but not by ₂ or -blockers. The results demonstrate that the ketogenic effect of norepinephrine is coupled with an inhibitory effect on lipogenesis which may be explained by diminished activity of acetyl-CoA carboxylase, diminished formation of malonyl-CoA and decreased activity of carnitine palmitoyl transferase I.     

On the binding of steroid sulfates to albumin

3H-Labeled steroid sulfates, sulfate of estrone (E1S) or dehydroepiandrosterone (DHAS), were dialyzed against delipidated human serum albumin or human plasma in the presence of increasing amounts of competing non-labeled sulfates (DHAS or E1S). The apparent equilibrium constants (K) of the tracers did not measurably change at concentrations of the non-radioactive sulfates below 10(-5) mol/l. At higher concentrations, K decreased gradually. The apparent equilibrium constant of 3H-E1S was diminished by plasma in a similar fashion. It may be concluded that albumin possesses one strong, non-specific binding site. This site, however, does not seem to be utilized for the binding of E1S in vivo, because of its preferential occupation by other ligands. This may be true for other steroid sulfates as well, depending on their relative abundance in plasma.     

Plating efficiency of mouse embryo cells as a function of gestational age

Summary Mouse embryo cells can be plated directly from single-cell suspensions of fresh embryos. For randomly bred Swiss mice, there is an increase in plating efficiency as a function of gestational age. Colony-forming units appear at about day 10 and there is an exponential increase in colony-forming ability up to day 16, after which there is no further increase.The technical assistance of Ms.Helen Sandza is gratefully acknowledged. This work was supported by a grant from the National Cancer Institute (No. CA13053).     

Lokale Kapillarpermeabilität und austauschbares Albumin in verschiedenen Organen der Ratte

Summary It was shown that the various organs and tissues differ from each other: (1) in the local capillary permeability for homologous I¹³¹-labeled serum albumin, and (2) in their contents of exchangeable albumin.     

Effect of albumin on uncoupling of oxidative phosphorylation by chinoform in rat liver mitochondria.

Addition of serum albumin diminished the uncoupling effect of chinoform on oxidative phosphorylation in rat liver mitochondria. Upon increasing the concentration of magnesium ions in the medium, the action of serum albumin was diminished. These results indicate that serum albumin combines with chinoform in competition with magnesium ions.     

Alpha 1-adrenergic stimulation of ketogenesis and fatty acid oxidation is associated with inhibition of lipogenesis in rat hepatocytes

The effect of norepinephrine on fatty acid synthesis (3H2O incorporation into fatty acids), on fatty acid oxidation to CO2 and on ketogenesis was studied in isolated hepatocytes of fed rats. After incubation with norepinephrine (50 microM), lipogenesis was lower (5.7 +/- 1.1 nmoles 3H2O incorporated into fatty acids/mg dry weight/30 min) than in controls (7.5 +/- 1.7; n = 6, p less than 0.02). In contrast, (1-14C) palmitate conversion into total ketone bodies was increased to 10.9 +/- 1.8 nmoles/mg/30 min with norepinephrine, vs 8.5 +/- 1.6 in controls (p less than 0.05), and more (1-14C) palmitate was converted to 14CO2 with norepinephrine than in controls (1.48 +/- 0.10 nmoles/mg/30 min vs 1.06 +/- 0.11, p less than 0.05). The inhibitory effect of norepinephrine on lipogenesis was abolished by addition of the alpha 1-receptor blocker prazosin, but not by alpha 2 or beta-blockers. The results demonstrate that the ketogenic effect of norepinephrine is coupled with an inhibitory effect on lipogenesis which may be explained by diminished activity of acetyl-CoA carboxylase, diminished formation of malonyl-CoA and decreased activity of carnitine palmitoyl transferase I.     

Effects of cadmium on the immune system of mice

Summary Chronic oral exposure of mice to Cd⁺⁺ inhibits cell-mediated immunity of delayed type hypersensitivity induced by sheep red blood cells (SRBC). No effect was detected on humoral immune response to SRBC. Spleen cells derived from mice exposed to Cd⁺⁺ showed in vitro enhanced response to T and B cell mitogens. These results demonstrate that Cd⁺⁺ exposure alters the immune system of mice.Dedicated to Prof. Georg Henneberg on the occasion of his 70th anniversary on 12.10.1978.Acknowledgments. Supported by a grant from the Umweltbundesamt. We thank Ms Odenwald, Ms Schulz, Klinikum Steglitz, and Ms Emeis, Robert Koch-Institut Abt. Immunologie, for the excellent technical assistance.     

Diffusion loading conditions determine recovery of protein synthesis in electroporated P3X63 Ag8 cells

Summary Using the suspension cell line P3X63 Ag8 we have studied the impact of the composition of the diffusion medium on cellular protein synthesis under standard electroporation conditions in TBS-Na. This buffer contains the high saline concentration usually present in electroporation-mediated DNA transfection. Electroporation in the presence of TBS-Na resulted in an immediate shut-off of protein synthesis, even though both FITC-dextran (M_r 40 kD) and Semliki Forest virus core protein (M_r 33 kD) were incorporated efficiently into the cytoplasm across the electropores at 0°C. Subsequent resealing of the pores was completed after a 5-min incubation at 37°C. When compared with control cells, overall protein synthesis of electroporated cells recovered slowly to resume a 30% activity after 1 h of incubation at 37°C. We have determined optimal conditions for diffusion loading (which necessitates the presence of ATP, GTP, amino acids, K⁺, Mg²⁺, and Ca²⁺) and resealing (in the presence of K⁺, Mg²⁺, and Ca²⁺), leading to a full and lasting recovery of protein synthesis within 5 min after pore closure.     

Antifertility activity of trimethylphosphate in male rats

Summary The toxic effect of trimethylphosphate on rat testes was investigated after oral dosing with 250 mg/kg for either 5 days/week for 30 days or 6 days/week for 60 days. After these prolonged treatments azoospermia resulted.Acknowledgments. The authors wish to thank Ms M. Michelles, Ms A. Murray and Ms R. Peake for their technical support.     

Chemical characterization of human urine albumin in proteinuria.

From the urine of a patient with proteinuria, the albumin protein component was isolated and compared with human serum albumin. By comparing the amino acid composition of the original proteins and their large cyanogen bromide fragments, peptide maps and N-terminal sequences of 33 amino acid residues, the identity of both proteins was shown.     

The chemokinetic effect of serum albumin

Summary Experiments performed by means of time lapse cinematography or the filter technique show that human serum albumin has marked chemokinetic effects on neutrophil cultured in Gey's solution. The average speed of the cells, as well as the proportion of neutrophils showing locomotion, is increased. Enhanced locomotion correlates with decreased attachment to the substratum as determined by morphological and functional criteria.We thank Mrs R. Graf and Miss M. Schuster for technical assistance. This work was supported by the Swiss National Science Fondation.     

Total replacement of blood by an emulsion of fluorocarbon in the rat-water extravasation as a cause of failure

Summary After exchange transfusion by an emulsion of fluorocarbons (Fluosol 43) in rats, an increase in fluorocarbons and¹³¹I-labelled albumin was observed. These changes suggest a transfer of water from vascular to interstitial space possibly owing to the inability of the emulsion to reproduce the oncotic pressure of normal blood.     

Plakorin,a potent Ca2+-ATPase activator from the Okinawan marine spongePlakortis sp.

Summary A new cyclic peroxide, plakorin, which is a potent sarcoplasmic reticulum (SR) Ca^a+-ATPase activator has been isolated from the Okinawan marine spongePlakortis sp., its structure was elucidated on the basis of spectral data.Acknowledgments. We thank Ms M. Hamashima and Ms A. Muroyama for their technical assistance.     

Symbioramide,a novel Ca2+-ATPase activator from the cultured dinoflagellateSymbiodinium sp.

Summary A novel sphingosine derivative, symbioramide, has been isolated from the laboratory-cultured dinoflagellateSymbiodinium sp. as a sarcoplasmic reticulum (SR) Ca²⁺-ATPase activator, and its structure elucidated to be1 on the basis of spectral and chemical means.Acknowledgments. We thank Ms M. Hamashima and Ms A. Muroyama for their technical assistance.     

Effects of selective dopamine D1 and D2 antagonists on male rat sexual behavior

Summary The effects of selective dopamine (DA) D₁ and D₂ antagonists on male rat sexual behavior were investigated. The D₁ antagonist (+)SCH-23390, 25–100 g kg^–1 s.c. –20 min, and the D₂ antagonist raclopride, 0.1–1.6 mg kg^–1 s.c., –20 min, decreased both the number of mounts and intromissions preceding ejaculation. No statistically significant effects in the time up to ejaculation or in the time up to the first intromission were noted, whereas both compounds produced a statistically significant increase in the post-ejaculatory interval. The effect can generally be characterized as psychomotor inhibition, and no evidence was obtained for a specific role of DA D₁ or D₂ receptors in the mediation of male rat sexual behavior.The expert technical assistance of Ms Elisabeth Wallin is gratefully acknowledged. The figures were skilfully prepared by Ms Madelene Kröning at the Department of Psychology. This study received support from the Bank of Sweden tercentenary Foundation, The Swedish MRC and Wilhelm and Martina Lundgren Foundation.