Acquisition of vimentin in astrocytes cultured from postnatal rat brain

Summary Vimentin and glial fibrillary acidic protein (GFAP) represent the principal constituents of intermediate filaments found in astrocytes. In contrast to vimentin—GFAP transition which occurs during glial developmentin situ, vimentin coexists with GFAP in cortical astrocytes allowed to differentiate in culture. To examine whether culture conditions or proliferative activity of the cells is responsible for the expression of vimentin, we generated cultures of GFAP-positive, vimentin-negative astrocytes isolated from 26-day postnatal rat brain cortices. Isolated astrocytes are characterized by a very thin rim of perinuclear cytoplasm and by numerous processes. Antiserum to GFAP labelled major processes and cell somata of some astrocytes, especially those with relatively short and large processes. Within 3 days in culture, all astrocytes accumulated GFAP in hypertrophic cell bodies and many began to express vimentin. Vimentin appeared primarily close to nuclei, and filaments of vimentin extended into proximal segments of the cell processes. In some astrocytes, however, vimentin was always absent. Combined double immunolabelling and histoautoradiography experiments demonstrated that the acquisition of vimentin was independent of the ability of astrocytes to incorporate tritiated thymidine. The results indicate that astrocytes isolated from 26-day postnatal rat brain are heterogeneous with respect to their ability to express vimentin and that vimentin synthesis is not correlated with the growth state of the cells as had been previously suspected.     

Our experience with diagnostics of congenital disorders of glycosylation

The aim of this study is to report our 3years experience with the screening of congenital disorders of glycosylation. A common isoelectric focusing method with immunofixation was used for analysis of serum transferrin and alpha1-antitrypsin, apart from several other procedures. A group of about 1000 individuals, both healthy controls and patients, mostly with signs of a metabolic disease were examined. Here we present an overview of (1) hypoglycosylation findings, (2) distribution of protein variants, (3) misguiding rare Tf variants found in our set, and (4) association of some phenotypes with various diseases.     

Polymerization of lactams, 55. Cyclic dimer of 6-caprolactam and its polymerization

Polymerization of the cyclic dimer of 6-caprolactam was investigated in N-methyl-2-pyrroli-done solution at 260°C under initiation with 6-aminocaproic acid or anhydrous phosphoric acid on the one hand, and in the solid phase at 230, 260, and 280°C in the absence of any other component or under initiation with anhydrous phosphoric acid, on the other. Monomeric 6-caprolactam was formed in all cases already at the beginning of reaction. It has been suggested that, in addition to the direct incorporation of the cyclic dimer into a polymer chain, the dimer is alos incorporated via the monomer, which is formed from the dimer by an intermolecular transamidation reaction.     

Structure and stability of poly(vinyl chloride), 4. Unsaturated ketones and their role in the initiation of thermal degradation

The thermal dehydrochlorination of poly(vinyl chloride) (PVC) in an inert atmosphere was investigated by the continuous potentiometric method. The effect of the structure of alicyclic and aliphatic α,β-unsaturated ketones on the rate of dehydrochlorination was examined, and a new mechanism of initiation for the dehydrochlorination of PVC was suggested. The results obtained demonstrate that the basic cause of the thermal instability of PVC are structures of the type of cisoid α,β-unsaturated ketone, dienone, or polyenone which by the dehydrochlorination of regular structural PVC units form isolated double bonds which, in turn, initiate the chain dehydrochlorination of PVC by allyl activation.     

The ankle-brachial blood pressure index as a risk indicator of generalized atherosclerosis

Contemporary methods of assessment of possible cardiovascular disease based on traditional risk factors are not perfect. Therefore new ways are sought. Simple and inexpensive methods include assessment of the blood pressure on the lower extremities above the ankles. The finding of a reduced ratio of the ankle-brachial blood pressure index (ABI) indicates atherosclerosis of the lower extremity arteries. ABI values of 0.9 or less are associated with a significantly increased risk of cardiovascular diseases (in particular myocardial infarction and stroke) that is independent of other risk factors. ABI assessment is a simple clinical examination and should be performed in middle-aged and elderly subjects with a medium and high cardiovascular risk in order to define more accurately their health status. Patients with reduced ABI of 0.9 or less are candidates for secondary preventive treatment for cardiovascular disease.     

Impact of azole drugs on energetics,kinetics, and ligand migration pathways of CO photo-dissociation in bacterial flavohemoglobins

Flavohemoglobins (fHbs) are heme proteins found in prokaryotic and eukaryotic microbes. They are involved in NO detoxification through an NO˙ dioxygenase mechanism. The N-terminal heme globin domain allows for binding of gaseous ligands whereas a C-terminal NADH/FADH binding domain facilitates association of redox cofactors necessary for ligand reduction. The NO˙ dioxygenase function is important in facilitating immune resistance by protecting the cell from nitrosative stress brought about by a host organism; as a result, bacterial flavoHbs have recently been considered as targets for the development of new antibiotics. Here, photoacoustic calorimetry and transient absorption spectroscopy have been used to characterize energetics, structural dynamics, and kinetics of CO migration within bacterial flavoHbs from Ralstonia eutropha (FHP) and Staphylococcus aureus (HMP_Sa) in the presence and absence of antibiotic azole compounds. In FHP, the ligand photo-release is associated with ΔH = 26.2 ± 7.0 kcal mol⁻¹ and ΔV = 25.0 ± 1.5 mL mol⁻¹ while in HMP_Sa, ΔH = 34.7 ± 8.0 kcal mol⁻¹ and ΔV = 28.6 ± 17 mL mol⁻¹ were observed, suggesting distinct structural changes associated with ligand escape from FHP and HMP_Sa. In the presence of ketoconazole, the CO escape leads to a more negative enthalpy change and volume change whereas association of miconazole to FHP or HMP_Sa does not impact the reaction volume. These data are in agreement with the computational results that propose distinct binding sites for ketoconazole and miconazole on CO bound FHP. Miconazole or ketoconazole binding to either protein has only a negligible impact on the CO association rates, indicating that azole drugs do not impact flavoHbs interactions with gaseous ligands but may inhibit the NOD activity through preventing the electron transfer between FAD and heme cofactors.

Impact of ketoconalzole and miconazole on structural dynamics of flavohemoglobin.     

Outcome and management of pregnancies in severe chronic neutropenia patients by the European Branch of the Severe Chronic Neutropenia International Registry

Long-term granulocyte-colony stimulating factor treatment has been shown to be safe and effective in severe chronic neutropenia patients. However, data on its use during pregnancy are limited. To address this issue, we analyzed all pregnancies reported to the European branch of the Severe Chronic Neutropenia International Registry since 1994. A total of 38 pregnancies in 21 women with chronic neutropenia (16 pregnancies in 10 women with congenital, 10 in 6 women with cyclic, 12 in 5 women with idiopathic neutropenia) were reported. Granulocyte-colony stimulating factor was administered throughout pregnancy in 16 women and for at least one trimester in a further 5 women. No major differences were seen between treated and untreated women with respect to pregnancy outcome, newborn complications and infections. In addition, we evaluated the genetic transmission of known or suspected genetic defects in 16 mothers having 22 newborns as well as in 8 men fathering 15 children. As a proof of inheritance, neutropenia was passed on to the newborn in 58% from female and in 62% from male patients with ELANE mutations, but also to some newborns from parents with unknown gene mutation. Based on our results, granulocyte-colony stimulating factor therapy has been shown to be safe for mothers throughout pregnancies and for newborns without any signs of teratogenicity. With an increasing number of adult patients, genetic counseling prior to conception and supportive care of mothers during pregnancy are crucial. The acceptance of having affected children may reflect the high quality of life obtained due to this treatment.     

Shape Differences Between the Faces of Homosexual and Heterosexual Men

Previous studies have shown that homosexual men differ from heterosexual men in several somatic traits and lay people accurately attribute sexual orientation based on facial images. Thus, we may predict that morphological differences between faces of homosexual and heterosexual individuals can cue to sexual orientation. The main aim of this study was to test for possible differences in facial shape between heterosexual and homosexual men. Further, we tested whether self-reported sexual orientation correlated with sexual orientation and masculinity–femininity attributed from facial images by independent raters. In Study 1, we used geometric morphometrics to test for differences in facial shape between homosexual and heterosexual men. The analysis revealed significant shape differences in faces of heterosexual and homosexual men. Homosexual men showed relatively wider and shorter faces, smaller and shorter noses, and rather massive and more rounded jaws, resulting in a mosaic of both feminine and masculine features. In Study 2, we tested the accuracy of sexual orientation judgment from standardized facial photos which were assessed by 80 independent raters. Binary logistic regression showed no effect of attributed sexual orientation on self-reported sexual orientation. However, homosexual men were rated as more masculine than heterosexual men, which may explain the misjudgment of sexual orientation. Thus, our results showed that differences in facial morphology of homosexual and heterosexual men do not simply mirror variation in femininity, and the stereotypic association of feminine looking men as homosexual may confound judgments of sexual orientation.