Distribution of macroinvertebrates in the shallow part of ‘De Gijster’, a water storage lake in the Biesbosch (The Netherlands)

The composition and distribution of the macroinvertebrate fauna of the shallow part of De Gijster, a medium sized water storage lake in The Netherlands, has been studied during 1984. The numerous individuals collected belong mainly to the Oligochaeta, Diptera (Chironomidae larvae) and Mollusca. Amongst them are elements of both the former polder fauna and the river fauna. A quantitative analysis revealed data on the horizontal distribution of several taxa within the lake as well as data on the influence of the lifecycle of some taxa on the annual composition of the fauna.     

Temporal Distribution of [3H]-Imipramine Binding in Rat Brain Regions is Not Changed by Chronic Methamphetamine

Specific binding of [³H]-imipramine in the rat suprachiasmatic nuclei, occipital cortex and caudate putamen underwent significant and replicable changes throughout 24 hr under a light-dark cycle or under constant conditions. Daily variations were also found in the medial and dorsal raphe nuclei and the lateral hypothalamus. Methamphetamine, a psychoactive drug with marked effect on circadian rhythms in physiological and hormonal parameters and adrenergic receptors, did not have any significant effect on imipramine binding rhythms in eight discrete brain regions. Thus a drug known to reduce serotoninergic neurotransmission did not change characteristics of the modulatory binding site related to serotonin uptake.     

Presence of glyoxalase II in mitochondria from spinach leaves: comparison with the enzyme from cytosol

Glyoxalase II has been purified from cytosol and mitochondria of spinach leaves. Electrophoresis and isoelectric focussing have resolved cytosolic and mitochondrial glyoxalase II in multiple forms: pl 5.3, 5.8 and 6.2 (cytosol) and pl 4.8 (mitochondria). The enzyme of both localizations is a monomer showing a relative molecular mass of about 26 kDa. The values of kinetic constants using several glutathione thiolesters as substrates, are similar for the enzymes from cytosol and mitochondria. These results extend also to plant the presence in mitochondria of peculiar forms of glyoxalase II, likewise recently demonstrated in mammalians.     

The induction of lincomycin resistance in Lycopersicon peruvianum and Lycopersicon esculentum

Lincomycin-resistant calli were induced from both Lycopersicon esculentum and Lycopersicon peruvianum using N-mitroso-N-methylurea (NMU) mutagenesis. From these calli lincomycin-resistant plants were regenerated. For L. peruvianum it was shown that the resistant plants could be divided in two classes with respect to their resistance to lincomycin and its derivative clindamycin. The first class comprised plants which were resistant to 500 mg/l lincomycin and showed no shoot or root formation in the presence of clindamycin; the second class consisted of plants resistant to 2000 mg/l lincomycin and these plants were able to form shoots and roots on clindamycin containing media. Lincomycin is an inhibitor of peptidyltransferase; chloroplast encoded parts of this enzymatic function are sensitive for this antibiotic. Reciprocal crosses between our lincomycin resistant and wild type L. peruvianum plants indicated a maternal inheritance of the mutation.     

Immunoaffinity Purification of Epitope-Tagged Human β2-Adrenergic Receptor to Homogeneity

To obtain large quantities of pure human β₂-adrenergic receptor (β₂-AR) needed for structural studies, an efficient method for β₂-AR purification was developed using a recombinant receptor with an eight amino acid epitope at its C-terminus. This epitope is recognized by KT3-monoclonal antibody. The epitope tagged β₂-AR was expressed in Sf9 cells with a specific activity of 5–20 pmol/mg of membrane protein. The epitope-tagged and wild-type receptors had identical ligand binding properties. The tagged receptor was solubilized using dodecyl-β-maltoside with a quantitative yield. Solubilized epitope-tagged receptors were partially purified by KT3-mAb immunoaffinity in 60–70% yield. Further purification of the receptors on an alprenolol-affinity column resulted in a homogenous preparation with an overall yield of >30%. The purified receptor was concentrated to >1 mg/ml without loss of ligand binding activity.     

Production of amylase(s) by Schwanniomyces castellii and Endomycopsis fibuligera

Schwanniomyces castellii and Endomycopsis fibuligera Produced extracellular amylase(s) when grown on various carbon sources and at different pH values. Both yeast species showed significant amylase synthesis in the presence of either maltose or soluble starch. On the other substrates tested (glucose, cellobiose, sucrose, trehalose, melezitose, raffinose, ethanol, glycerol) differences were found regarding growth and amylase production. Free glucose in the culture medium apparently inhibited enzyme synthesis. The pH range allowing maximal growth and amylase production was 4.5–6.0 for E. fibuligera and 5.5–7.0 for S. castellii.     

Trisomy 6p22→6pter due to familial t(6;13)(p22;q34 or 33) translocation

Summary 235 cases of Down's syndrome were ascertained in a 10-year study of Down's syndrome in Western Australia. Although cytogenetic studies performed on 222 subjects confirmed that 95% of cases were trisomic due to nondisjunction, 4% were trisomic due to translocation, and 1% were mosaic, the ratio of inherited/sporadic translocations differed from that usually reported. Comparison of the results with those of an earlier Australian survey of Down's syndrome demonstrated a real fall in the incidence of Down's syndrome in Australia but no significant change in maternal age-specific incidences.     

Early midcell localization of Escherichia coli PBP4 supports the function of peptidoglycan amidases

Insertion of new material into the Escherichia coli peptidoglycan (PG) sacculus between the cytoplasmic membrane and the outer membrane requires a well-organized balance between synthetic and hydrolytic activities to maintain cell shape and avoid lysis. Since most bacteria carry multiple enzymes carrying the same type of PG hydrolytic activity, we know little about the specific function of given enzymes. Here we show that the DD-carboxy/endopeptidase PBP4 localizes in a PBP1A/LpoA and FtsEX dependent fashion at midcell during septal PG synthesis. Midcell localization of PBP4 requires its non-catalytic domain 3 of unknown function, but not the activity of PBP4 or FtsE. Microscale thermophoresis with isolated proteins shows that PBP4 interacts with NlpI and the FtsEX-interacting protein EnvC, an activator of amidases AmiA and AmiB, which are needed to generate denuded glycan strands to recruit the initiator of septal PG synthesis, FtsN. The domain 3 of PBP4 is needed for the interaction with NlpI and EnvC, but not PBP1A or LpoA. In vivo crosslinking experiments confirm the interaction of PBP4 with PBP1A and LpoA. We propose that the interaction of PBP4 with EnvC, whilst not absolutely necessary for mid-cell recruitment of either protein, coordinates the activities of PBP4 and the amidases, which affects the formation of denuded glycan strands that attract FtsN. Consistent with this model, we found that the divisome assembly at midcell was premature in cells lacking PBP4, illustrating how the complexity of interactions affect the timing of cell division initiation.     

Novel Surface Display System for Proteins on Non-Genetically Modified Gram-Positive Bacteria

A novel display system is described that allows highly efficient immobilization of heterologous proteins on bacterial surfaces in applications for which the use of genetically modified bacteria is less desirable. This system is based on nonliving and non-genetically modified gram-positive bacterial cells, designated gram-positive enhancer matrix (GEM) particles, which are used as substrates to bind externally added heterologous proteins by means of a high-affinity binding domain. This binding domain, the protein anchor (PA), was derived from the Lactococcus lactis peptidoglycan hydrolase AcmA. GEM particles were typically prepared from the innocuous bacterium L. lactis, and various parameters for the optimal preparation of GEM particles and binding of PA fusion proteins were determined. The versatility and flexibility of the display and delivery technology were demonstrated by investigating enzyme immobilization and nasal vaccine applications.