Carbohydrate receptor specificity of K99 fimbriae of enterotoxigenicEscherichia coli

K99 Fimbriae from enterotoxigenicEscherichia coli (ETEC) were found to bind specifically to sialic acid, as measured in a haemagglutination inhibition assay using the intact bacteria and human erythrocytes. The affinity forN-glycolylneuraminic acid was about twice that ofN-acetylneuraminic acid (NeuAc), and other monosaccharides were found to be at least ten-fold less effective as inhibitors. The specificity was found to depend on electrostatic interaction where the carboxyl group and its orientation plays an important role. 2--Benzyl-NeuAc was a better inhibitor than 2--methyl-NeuAc suggesting a hydrophobic patch near the binding site on the protein. Axially oriented hydroxyl groups as in 4-epi-NeuAc and 3-hydroxy-NeuAc seemed to participate in binding since these derivatives were better inhibitors thanN-acetylneuraminic acid. K99 was found to have a higher affinity for 4-O-acetyl-NeuAc and lower affinity forN-acetylneuraminic acid withO-substituents at C7-C9 as compared toN-acetylneuraminic acid. Hence, the degree ofO-acetylation of sialic acid in the mucosa of the small intestine may influence colonization and determine susceptibility to infection.     

The effect of monensin on thyroglobulin secretion

The effect of monensin on the secretion of thyroglobulin was studied in open follicles isolated from pig thyroid tissue; in this system, thyroglobulin is secreted into the incubation medium. When monensin was present during a 4-h chase incubation after pulse-labelling with 3H-leucine, the secretion of labelled thyroglobulin was reduced by about 85%; in electron-microscopic autoradiographs of rat thyroid lobes labelled and chase-incubated under similar conditions the relative number of grains over follicle lumina was strongly reduced when monensin was present during the chase. These observations are in agreement with the consensus that monensin arrests transport of secretory proteins in the Golgi complex. In other experiments, pulse-labelled follicles were chase-incubated for 1.5 h whereby labelled thyroglobulin was transported from the RER to exocytic vesicles. Monensin present during a subsequent chase of 0.5 h caused only a moderate decrease of labelled thyroglobulin secretion. TSH present during the second chase-stimulated secretion in both control and monensin-exposed follicles. TSH also caused a drastic reduction of exocytic vesicles in rat thyroid lobes, and the number of vesicles remaining in the cells was the same in controls and lobes exposed to the ionophore. The observations are interpreted to show that monensin does not inhibit the basal or TSH-stimulated transport of thyroglobulin from the site of monensin-induced arrest in the Golgi complex to the apical cell surface or the exocytosis of thyroglobulin.     

Testing the “methanochondrion concept”: are nucleotides transported across internal membranes in Methanobacterium thermoautotrophicum?

In order to test the Methanochondrion concept, uptake of adenine nucleotides in various membrane preparations of Methanobacterium thermoautotrophicum was studied. The uptake showed properties which are in general interpreted as indicative of a transport mechanism: (i) kinetics in the time range of minutes, (ii) temperature dependence, (iii) substrate specificity and (iv) failure to remove the substrate by extensive washing.However, nucleotide transport as an interpretation of this uptake can definitely be excluded. Not only an exchange mechanism of the mitochondrial type, but also a general exchange or an uniport mechanism was ruled out. In contrast, the nucleotide uptake was shown to be actually a tight and specific binding of ADP and ATP to binding sites at the interior side of the cell membrane. This was conclusively demonstrated in protoplasts obtained from M. thermoautotrophicum cells. In these protoplasts which do not contain internal membranes also nucleotide binding was observed, but only after disruption of the plasma membrane by osmotic lysis, which leads to the exposure of binding sites.     

Chromosome analysis of human sperm

Summary A modified technique has been developed for the visualization of the chromosomes in human sperm. The cytogenetic analysis of 129 G-banded human sperm metaphases of 6 normal donors showed an incidence of structural and numerical chromosome abnormalities of 7.8%. Two out of 129 spermatozoa were aneuploid (1.6%). The frequency of sperms with chromatid-type aberrations was 2.3% (3/129). Chromosome-type aberrations were found in 5 out of 129 (3.9%) spermatozoa. X to Y ratio did not differ significantly from the expected one-to-one ratio. Twenty-six sperm complements from a patient 18–20 months after testes exposure to 30 Gy were examined. A significant increase of numerical and structural chromosome abnormalities was not observed. Chromatidtype aberrations were found in two sperm complements (7.7%) and chromosome-type aberrations in one sperm complement (3.9%). The cytogenetic analysis of 15 human sperms from a cancer patient 26 months after chemotherapy showed an increased frequency of aberrant sperm complements (33.4%). One chromatid-type (6.7%), three chromosometype aberrations (20.0%) and one (6.7%) hyperploid sperm complement could be observed. The sample size is still too small to answer the question whether chemical mutagens may increase the frequency of chromosomal abnormalities in human sperm.     

Microinjection of normal cell extracts into Fanconi anemia fibroblasts corrects defective scheduled DNA synthesis recovery after 8-methoxypsoralen plus UVa treatment

Summary Fanconi anemia (FA) cells show an increased sensitivity to 8-methoxypsoralen (8-MOP) plus UVa treatment; after an initial reduction of their semiconservative DNA synthesis rate, they do not recover like normal cells. We microinjected extracts from normal cells into FA fibroblasts from complementation group A and determined semiconservative DNA synthesis rates by autoradiography; the hampered recovery phase was completely restored.The present data are part of M. Gök's thesis 1987 at the University of Heidelberg     

Diagnosis of genetic disease using recombinant DNA. Supplement

Summary Recombinant DNA methodology has greatly increased our knowledge of the molecular pathology of the human genome at the same time as providing the means to diagnose inherited disease as the DNA level. We present here a list of recent reports of both direct and indirect analysis of human inherited disease which is intended to serve as a guide to current molecular genetic approaches to diagnostic medicine.     

Effects of N-immissions on nutrient status and vitality ofPinus sylvestris near a hen-house

Near a hen house (50–600 m), vitality ofPinus sylvestris, N-, P-, K-, Ca-, Mg-contents of the needles, N-, Mg-, K-, Ca- and Al-contents in soil extracts and NH₃/NH ₄ ⁺ -contents of the air were determined. Damage symptoms occurred when N-immissions hit the canopy directly. In contrast no visible decline of the above ground plant could be observed if N was mainly deposited on the soil.     

Light-induced spectral absorbance changes in relation to photosynthesis and the epoxidation state of xanthophyll cycle components in cotton leaves

When cotton (Gossypium hirsutum L., cv Acaia SJC-1) leaves kept in weak light were suddenly exposed to strong red actinic light a spectral absorbance change took place having the following prominent characteristics. (a) It was irreversible within the first four minute period after darkening. (b) The difference in leaf absorbance between illuminated and predarkened leaves had a major peak at 505 nanometers, a minor peak at 465 nanometers, a shoulder around 515 nanometers, and minor troughs at 455 and 480 nanometers. (c) On the basis of its spectral and kinetic characteristics this absorbance change can be readily distinguished from the much faster electrochromic shift which has a peak at 515 nanometers, from the slow, so-called light-scattering change which has a broad peak centered around 535 nanometers and is reversed upon darkening, and from absorbance changes associated with light-induced chloroplast rearrangements. (d) The extent and time course of this absorbance change closely matched that of the deepoxidation of violaxanthin to zeaxanthin in the same leaves. (e) Both the absorbance change and the ability to form zeaxanthin were completely blocked in leaves to which dithiothreitol (DTT) had been provided through the cut petlole. DTT treatment also caused strong inhibition of that component of the 535-nanometer absorbance change which is reversed in less than 4 minutes upon darkening and considered to be caused by increased light scattering. Moreover, DTT inhibited a large part of nonphotochemical quenching of chlorophyll fluorescence in the presence of excessive light. However, DTT had no detectable effect on the photon yield of photosynthesis measured under strictly rate-limiting photon flux densities or on the light-saturated photosynthetic capacity, at least in the short term. We conclude that it is possible to monitor light-induced violaxanthin de-epoxidation in green intact leaves by measurement of the absorbance change at 505 nanometers. Determination of absorbance changes in conjunction with measurements of photosynthesis in the presence and absence of DTT provide a system well suited for future studies of meachanisms of dissipation of excessive excitation energy in intact leaves.