Serum cholinesterase: function in lipoprotein metabolism.

Human serum beta-lipoproteins, isolated by percipitation with heparin-calcium mixture, showed cholinesterase activity. The enzyme activity was almost proportional to the lipoprotein concentration. Rats, treated with neostigmine, a cholinesterase inhibitor, showed a significant decrease in serum beta-lipoprotein and in the incorporation of H3-lysine into the lipoprotein compared to untreated controls. The decreased incorporation of H3-lysine into beta-lipoprotein was associated with increased labelling of alpha-lopoprotein. There was no significant difference in the labelling of pre-beta-lipoprotein. We propose that LDL is formed from VLDL in the presence of cholinesterase.     

Cholinesterase in the atherosclerotic intima and in fibroblast cultures

Summary Cholinesterase activity was present in the atheromatous plaque of the rabbit's atherosclerotic aorta. Cholinesterase activity was significantly increased in rat fibroblast cultures grown in the presence of hypercholesterolemic serum. Cholesterol ester synthesis in these cultures was inhibited by neostigmine, a cholinesterase inhibitor.Acknowledgments. We thank Patricia Fontaine for technical assistance and Patricia Candow and Barbara English for secretarial help. This work was supported by a grant from the Canadian Heart Foundation. Part of this work was presented at 1977 annual meeting of the Biochemical Society in London.     

The growth responses of hamster chondrocytes,dermal fibroblasts and embryo cells to whole blood serum and plasma-derived serum

Summary Autoradiographic studies with³H-thymidine demonstrated that the growth responses of hamster chondrocytes, dermal fibroblasts and embryo cells, respectively, differed in media containing whole blood serum (WBS) and plasmaderived serum (PDS). Dermal fibroblasts seemed to require a growth factor from platelets for growth, but chondrocytes did not. Embryo cells showed an intermediate pattern of growth response to this factor.This work was supported by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science and Culture and the Ministry of Health and Welfare of Japan.We thank Miss M. Tanaka and Miss K. Kawana for technical assistance.     

Mg2+-ATPase defective mutant ofEscherichia coli and thiamine transport

Summary Mg²⁺-ATPase deficient mutant ofEscherichia coli showed an evident dependency of thiamine uptake on the oxidative metabolism of glucose, whereas the parent strain did not. In both cells, this uptake was completely inhibited by H⁺ conductors.Acknowledgment. We are indebted to Miss M. Abe for her excellent technical assistance.     

Effect of low density lipoprotein on proteoglycan synthesis by aorta cells in culture

Summary Increasing the content of human serum low density lipoprotein in the growth medium led to greater incorporation of³⁵S-sulfate into proteoglycan (mostly into dermatan sulfate) by primary aorta cells but did not affect similar incorporation by fibroblast cells. These results suggest a mechanism which can explain the increased deposition of lipid in aorta due to hyperlipidemia.Acknowledgements. This work was supported by a Fort Polk-American Heart Association-Louisiana, Inc. Research Award.     

Effects of acute intoxication with uranyl nitrate on bone formation

Summary The alteration of bone formation by an acute intoxication with uranyl nitrate is demonstrated by histologic and histometric methods. When compared with the controls, intoxicated animals showed a markedly lower density in healing sockets, while bone formation was reduced in healing sockets as well as in metaphyseal bone.Acknowledgments. This work was supported in part by grant No. 8461 d/82 of the National Research Council Argentina and by Research Contract No. 2816/R1/R2 of the International Atomic Energy Agency. The authors wish to acknowledge the valuble technical assistance of Miss Ofelia Chiesa and Miss Patricia Mandalunis.     

Dose-effects of 8-methoxypsoralen and long wave UV-light in 3T3 cells: evaluation of a phototoxic index

Summary 3T3 cells were cultured until confluency and treated with various doses of 8-methoxypsoralen followed by long wave UV light irradiation. The inhibition of³H-thymidine incorporation was dose-dependent for both, psoralen and light. A phototoxic index (PTI) was established demonstrating that a constant correlation between psoralen and UVA light exists for the photoinactivation in living cells.Supported by Deutsche Forschungsgemeinschaft.We thank Miss Anne Schröder for her skillful assistance.     

Biliary excretion of sulfobromophthalein in isolated perfused livers from normal and spironolactone-treated rats

Summary Bile flow and biliary excretion of sulfobromophthalein (BSP) was examined in isolated perfused livers from normal and spironolactone(SP)-treated rats. BSP biliary excretion contributed to the bile production in both groups. Moreover SP increased BSP biliary excretion but transfer of dye from plasma into liver was not affected.Acknowledgments. This work was supported by a grant from Consejo Nacional de Investigaciones Científicas y Técnicas República Argentina (EFIHER-CONICET). Isolated rat liver preparations were performed by Miss Graciela Patricia Rodriguez. The assistance of Carlos Guillermo Grieve is also gratefully acknowledged.     

Changes in cholinesterase activity of muscle after crushing the sciatic nerve of rats

Summary The subcellular distribution of cholinesterase (ChE) was studied in the gastrocnemius muscle of rats after strong or weak nerve crushing. The ChE activities of muscle were decreased to a greater extent by strong crushing than by weak crushing. In particular, the ChE activity of the fraction containing sarcoplasmic reticulum was most greatly decreased. These results suggest that the change in the ChE activity of the microsomal fraction most finely reflects the strength of nerve crushing.Acknowledgments. This study was accomplished in Central Research Laboratories, Sankyo Co. Ltd. The authors wish to thank Prof. Tsuneyuki Nakazawa, Department of Neuropsychiatry, School of Medicine, Fujita-Gakuen University, and Dr Yutaka Sakai for their valuable advice and support, Sr Jean M. Michalec for her critical comments, and Miss Hamako Katano, Mr Naoji Mikuni and Miss Yoshie Ishii for their skillful technical assistance.     

Effect of intralipid infusion on serum high- and low-density lipoprotein cholesterol,lecithin: Cholesterol acyltransferase,and lipoprotein lipase in tumor-bearing rats

We compared the effects of 0.45% normal saline (NS), 5% Intralipid^® (IL), and 16.7% glucose (Glu) infusions on total serum triglycerides and cholesterol, serum high-(HDL-c) and low-density lipoprotein cholesterol (LDL-c), and activity of serum lecithin: cholesterol acyltransferase (LCAT), and serum lipoprotein lipase (LPL) in rats implanted with a fibrosarcoma. In tumor-bearing rats given NS, a two-fold increase in total serum cholesterol, a four-fold increase in LDL-c, and a five-fold decrease in the HDL-c/LDL-c ratio were observed compared to tumor-free rats. In tumor-bearing rats administered IL, a two-fold increase in total serum triglyceride and cholesterol, a three-fold increase in HDL-c and HDL-c/LDL-c ratio, and a two-fold increase in LPL activity were observed compared to tumor-bearing rats administered NS. In tumor-bearing rats administered Glu, a two-fold decrease in total serum cholesterol, a two-fold decrease in HDL-c, and a three-fold decrease in LDL-c were observed compared to tumor-bearing rats administered NS. Tumor weights and LCAT activity did not differ significantly between treatment groups. Previous results have demonstrated that lipophilic compounds that interact with plasma lipoproteins have altered pharmacological effects when administered with IL. Therefore, this study suggests that IL infusions alter the HDL-c/LDL-c ratio and could affect the pharmacological behavior of anticancer compounds that predominantly distribute into the LDL fraction upon entrance into the bloodstream.     

Cyclic nucleotide levels in the perfused rat heart subjected to hypoxia

Summary Isolated rat hearts were subjected to hypoxic perfusion on a recirculating Langendorff apparatus. Following a 30-min-period of aerobic stabilization the hearts were perfused for 30 min with media equilibrated with 84% N₂, 12% O₂ and 4% CO₂. At the end of the hypoxic period myocardial concentrations of cyclic AMP and cyclic GMP were determined by radioimmunoassay. Exposure to hypoxia resulted in a significant increase in cyclic AMP (p<0.01) and a decrease in cyclic GMP (p<0.05) as compared to hearts perfused for 60 min with media gassed with 96% O₂, 4% CO₂.Acknowledgment. We gratefully acknowledge the excellent technical assistance provided by Mrs Inger O. Boggs and Miss Patricia C. Hannigan. This work was supported in part by U. S. Public Health Service grant HL 14661 from the National Heart and Lung Institute, and by a grant from the Central Ohio Heart Chapter.     

Cholinesterases are down-expressed in human colorectal carcinoma

The aberrations of cholinesterase (ChE) genes and the variation of ChE activity in cancerous tissues prompted us to investigate the expression of ChEs in colorectal carcinoma. The study of 55 paired specimens of healthy (HG) and cancerous gut (CG) showed that acetylcholinesterase (AChE) activity fell by 32% and butyrylcholinesterase (BuChE) activity by 58% in CG. Abundant AChE-H, fewer AChE-T, and even fewer AChE-R and BuChE mRNAs were observed in HG, and their content was greatly diminished in CG. The high level of the AChE-H mRNA explains the abundance of AChE-H subunits in HG, which as glycosylphosphatidylinositol (GPI)-anchored amphiphilic AChE dimers (G₂^A) and monomers (G₁^A) account for 69% of AChE activity. The identification of AChE-T and BuChE mRNAs justifies the occurrence in gut of A₁₂, G₄^H and PRiMA-containing G₄^A AChE forms, besides G₄^H, G₄^A and G₁^H BuChE. The down-regulation of ChEs might contribute to gut carcinogenesis by increasing acetylcholine availability and overstimulating muscarinic receptors. Received 19 May 2006; received after revision 5 June 2006; accepted 5 July 2006     

Radioimmunoassay of some hormones simultaneously measured in serum and breast cyst fluid

Summary Blood and breast cyst fluid were drawn simultaneously for hormonal determination. There was no difference between serum and cyst fluid values of PRL and TSH. A significant difference was noted for LH (p<0.01) and FSH (p<0.05), serum concentrations being higher than cyst fluid concentrations.The authors are indebted to Miss J. Meister, E. Dullaert and J. Beck for their skillful technical help. We are also indebted to the National Institutes of Health, NIAMDD and the World Health Organization, International Laboratory for Biological Standards.     

Reassociation of eukaryotic ribosomal subunits by a factor from rat ascites hepatoma cytosol

Summary Post-ribosomal supernatant extracts from Yoshida AH 130 ascites hepatoma cells promote the in vitro association of ribosomal subunits at low Mg²⁺ concentration. Comparable extracts from rat liver show, on the contrary, dissociation factor activity on ribosome monomers.The technical assistance of Miss M. Ravazzani and Miss L. Zingaretti is gratefully acknowledged.     

Biological activity of flucycloxuron,a novel benzoylphenylurea derivative,onTenebrio molitor: comparison with diflubenzuron and triflumuron

Flucycloxuron, a novel benzoylphenylurea (BPU) derivative, exhibited insecticidal activity when injected into newly ecdysed pupae ofTenebrio molitor. Mortality occurs because of defective adult ecdysis. Treatment caused a reduction in both cuticle thickness and incorporation of¹⁴C-labelled precursor into chitin, although it had no significant effect on the protein synthesis. The potencies of other BPU compounds as inhibitors of chitin biosynthesis have been examined and results showed that diflubenzuron was less effective than either flucycloxuron or triflumuron.     

Different marrow cell number requirements for the haemopoietic colony formation and the cure of the W/Wv anemia

Summary The lowest cell number in the normal marrow transplant, which allows the cure of W/W^v anemia was found to be between 10⁴ and 10⁵. This exceeds by several times the lowest cell number necessary for the haemopoietic colony formation. Therefore, either the colony forming cell is not the haemopoietic stem cell but rather its progeny, or this cell requires an aid from some other cells to exert its activity.Acknowledgments. We are indebted to Doc. Dr Maksymilian Siekierzynski for his help and advice and to Miss Elzbieta Szewczyk for expert technical assistance.     

Effect of low density lipoprotein on proteoglycan synthesis by aorta cells in culture.

Increasing the content of human serum low density lipoprotein in the growth medium led to greater incorporation of 35S-sulfate into proteoglycan (mostly into dermatan sulfate) by primary aorta cells but did not affect similar incorporation by fibroblast cells. These results suggest a mechanism which can explain the increased deposition of lipid in aorta due to hyperlipidemia.     

Sodium and calcium action potentials in human anterior pituitary cells

Summary Human anterior pituitary cells derived from a somatotropin-secreting adenoma were capable of generating action potentials with Ca²⁺ and tetrodotoxin-sensitive Na⁺ components. A major fraction of the action current was carried by Na ions.We wish to thank Dr O. Sand for correcting the English and Miss Michiko Takano for technical assistance. We also acknowledge the collaboration of Dr S. Miyazaki in the electron microscope.     

Inhibition of Ca2+-induced noradrenaline release from central noradrenergic neurons by morphine

Summary Morphine inhibited the noradrenaline release from slices of rat brain cortex induced by introduction of Ca²⁺ ions after superfusion with Ca²⁺-free, K⁺-rich solution. The degree of inhibition was inversely related to the Ca²⁺ concentration used for stimulation.Acknowledgment. We thank Mrs G. Thielecke and Miss G. Werthmann for technical assistance.     

Net ATP synthesis by running the red cell calcium pump backwards

Summary Ca²⁺ loaded inside-out vesicles from human red blood cells, yielding Ca²⁺ 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.We are obliged to Dr H. Fey and Miss H. Pfister (Veterinarybacteriological Institute Bern) and Dr H. Porzig (Pharmacological Institute Bern) for help and advice.