Human arterial smooth muscle cells in culture: Inverse relationship between proliferation and expression of contractile proteins

Summary Human arterial smooth muscle cells (hASMC) from explants of the inner media of uterine arteries were studied in secondary culture. We had previously found that these cells depend on exogenous platelet-derived growth factor (PDGF) for proliferation in vitro. Deprivation of the serum mitogen(s) by culture in plasma-derived serum or bovine serum albumin (BSA) caused a true growth arrest that was reversible upon reexposure to the mitogen(s). When added to serum-containing medium, heparin caused a reversible growth arrest which could be competed for by increasing concentrations of serum. In the current study we used a set of smooth muscle-specific actin and myosin, antibodies to study the expression of contractile proteins in stress fibers under indirect immunofluorescence on hASMC in culture. Even in sparse culture, grwoth-arrested hASMC expressed stress fibers containing these actin and myosin epitopes. This was true irrespective of whether growth arrest was achieved by culture in media containing only BSA or a combination of heparin and whole blood serum. hASMC proliferating in whole blood serum in sparse culture did not express such strees fibers, as judged by immunofluorescent staining. This was true also for cells that were restimulated to proliferate in serum after a growth arrest. Utilizing a monoclonal antibody against a nuclear antigen expressed in proliferating human cells, we were able to demonstrate an inverse relationship between the expression of this antigen and the SMC-specific contractile proteins, respectively. Under these culture conditions, the reversible transition between defifferentiated and differentiated hASMC was almost complete and terminated about 1 wk after the change in culture condition. We conclude that hASMC in vitro respond, to exogenous PDGF by proliferation and dedifferetiation as a single population of cells. We also conclude that this modulation is reversible, because the cells become uniformly quiescent and differentiated when the mitogenic stimulus is blocked or removed. This study was supported by grants from the Swedish Medical Research Council (Project no. 4531 and 6816), the Swedish Association against Heart and Chest Diseases, the King Gustaf V and Queen Victoria Foundation, the National Institutes of Health, Bethesda, MD (grant HL 29873) and the Swedish National Board for Laboratory Animals.     

Simulated and measured nitrogen conditions in a manured and fertilised soil

The governing factors for soil nitrogen dynamics were identified with a simulation model. In addition, the model was used to interpret measurements from a plot fertilisation experiment in southwest Sweden.Simulated moisture and temperature conditions were the driving variables for the simulation of soil nitrogen dynamics and leaching during a 6-year period. The results of the simulation were compared with monthly observations on two plots with grain crops, one with liquid manure and commercial fertilisers applied and one with commercial fertilisers only.Simulated temporal variations of the nitrate and ammonium storages generally agreed with observations. The dominant role of the crops as a determinant of soil nitrogen conditions was demonstrated. A higher leaching loss from the plot with application of commerical fertilisers only occurred both in simulations and measurements compared to the plot with application of both commercial fertilisers and manure. The main reason was the higher N-application in the former treatment.The effect of water flows in macropores was interpreted as a delay of simulated leaching compared to observed leaching on some occasions in summer and early autumn. No direct effect of the macropores on the yearly rates of leaching could be seen.     

Selenium concentrations in the human thyroid gland

Recently, we found that prediagnostic serum selenium concentration was significantly lower for cases developing thyroid cancer (n=43) than for controls. We assumed that redistribution of serum selenium into the affected tissue took place in the prediagnostic period. The present study was carried out to determine the physiological concentration of selenium in the thyroid, since very few data are available in the literature. The concentrations of selenium in the thyroid (n=45) and liver samples from Norwegians who had died because of acute illness or accidents were determined by hydride generation atomic absorption spectrometry. The mean selenium concentration was found to be 0.72±0.44 μg/g in the thyroid and 0.45±0.11 μg/g in the liver tissue. The surprisingly high concentration of selenium in apparently normal thyroids indicates that selenium has important functions in this organ. The remarkably broad range, together with the observation that no significant correlation exists between thyroid and liver concentrations, suggest that factors other than the selenium status are important determinants for the selenium concentration in the thyroid gland. This observation is consistent with our hypothesis that in carcinogenesis, prediagnostic processes influence the serum-/thyroid-ratio of selenium.     

Temporal and spatial development of individual species of plankton algae from European lakes

The temporal and spatial developments of 15 phytoplankton species widely distributed in European and North American lakes, have been studied by informations in the literature and especially by their behaviour during a 5 years development in the Danish lake Grane Langsø. The species were characterized by temporal differences in maximum abundance, the density at the maximum, the number of maxima per year, the length of the vegetative period and the constancy of the annual maximum based on long-term studies. In several cases the phytoplankton species can be characterized by one individual behaviour in one group of lakes, while the same species behaves differently in another group of lakes. The results suggest that several species include two or more types with different tolerances and environmental requirements. Most of the species included are Chrysophytes, mainly species of Uroglena and Dinobryon. A remarkable feature of Grane Langsø's Chrysophytes is their predilection to accumulate in the bottom water during the summer. A similar behaviour was not demonstrated by Chrysophytes in other lakes.     

A remarkable new Bulbophyllum, sect. Sestochilus (Orchidaceae) from Thailand

Ongoing studies show that what hitherto in Thailand and neighboring countries have been gathered under one variable species, Bulbophyllum lobbii , must be separated in a number of distinct, well defined species. One such. B. smitinandii , is treated here.     

Determination of 4,4′-methylenediphenyldianiline (MDA) and identification of isomers in technical-grade MDA in hydrolysed plasma and urine from workers exposed to methylene diphenyldiisocyanate by gas chromatography-mass spectrometry

Gas chromatography-mass spectrometry using chemical ionization with ammonia as reagent gas monitoring both positive and negative ions was applied. Negative-ion monitoring using ammonia and the pentafluoropropionic anhydride (PFPA) derivatives were chosen owing to low detection limits and good separation for the isomers studied. Technical-grade methylenediphenyldiioscyanate (MDI) was analysed and three isomers, 4,4′−, 2.4′− and 2,2−methylenediphenyldianiline (MDA), were determined in addition to methylated MDA. Plasma and urine from an exposed worker were hydrolysed and analysed and the MDA isomers were identified in the biological samples.     

Four cytosolic-type CuZn-superoxide dismutases in germinating seeds of Pinus sylvestris

A differential analysis of CuZn-superoxide dismutase (SOD. EC 1.15.1.1) isozymes after native-polyacry lamide gel elecrrophoresis (PAGE) and isoelectric focusing (IEF) indicated that germinating seeds of Scots pine (Pinus sylvestris L.) 3 days after the start of imbibition (3 DAI) contain five CuZn-SOD isozymes. Two isozymes co-migrated on native–PAGE but were separated after IEF. CuZn-SODs of Scots pine were purified from germinating seeds (3 DAI) by anion-exchange chromatography, hydrophobic interaction chromatography and chromatofocusing. The final separation of CuZn-SOD isozymes was accomplished by native-PAGE. CuZn-SOD isozymes were electroblotted and their NH₂-terminal amino acid sequence was determined. Comparisons of the amino acid sequences with sequences of CuZn-SOD isozymes from other plant sources indicated that one CuZn-SOD isozyme was of the chloroplastic type whereas the other four isozymes belonged to the cytosolic-type CuZn-SODs, The NH₂-terminal amino acid sequence of the chloroplastic CuZn-SOD and of one cytosolic-type CuZn-SOD were identical to those of two previously isolated, sequenced and localized CuZn-SOD isozymes from Scots pine needles. Two cytosolic-type CuZn-SOD isozymes showed a homology at 20 out of 21 NH₂-terminal amino acids. Mitochondria and glyoxysomes were isolated by differential and Percoll density-gradient centrifugation from germinating seeds (3 DAI). The cell fractionation experiments did not suggest that a major part of the CuZn-SOD activity in germinating seeds was derived from glyoxysomes or mitochondria.     

High-temperature gas chromatography and gas chromatography-mass spectrometry of glycoprotein and glycosphingolipid oligosaccharides

High-temperature gas chromatography and gas chromatography-inass spectrometry for the analyses of oligosaccharides derived from glycoproteins or glycosphingolipids has been developed. Pcrmethylatcd oligosaccharides with up to about 12 sugar residues and masses up to 2500 Daltons can be analyzed. This approach is discussed and exemplified.