Penicillinamidohydrolase inEscherichia coli

Substrate specificity of the bacterial penicillinamidohydrolase (penicillinacylase, EC 3.5.1.11) fromEscherichia coli was determined by measuring initial rates of enzyme hydrolysis of different substrates within zero order kinetics. SomeN-phenylacetyl derivatives of amino acids and amides of phenylacetic acid and phenoxyacetic acid of different substituted amides of these acids or amides, structurally and chemically similar to these compounds, served as substrates. Significant differences in ratios of initial Tates of the enzyme hydrolysis of different substrates were found when using a toluenized suspension of bacterial cells or a crude enzyme preparation, in spite of the fact that the enzyme is localized between the cell wall and cytoplasmic membrane, in the so-called periplasmic space.N-phenylacetyl derivatives are the most rapidly hydrolyzed substrates. Beta-phenylpropionamide and 4-phenylbutyramide were not utilized as substrates. The substrate specificity of the enzyme is discussed with respect to a possible use of certain colourless compounds as substrates, hydrolysis of which yields chromophor products suitable for a simple and rapid assay of the enzyme activity.     

Biochemical dehydrogenations of saccharides

Fermentation of a medium containing 5% 2-deoxy-D-glucose and barium carbonate by a strain ofPseudomonas aeruginosa yielded barium 2-deoxy-d-gluconate. The yield was 77% theoretical. The strain in question makes it possible to prepare directly calcium, magnesium, manganese and ferrous salts of 2-deoxy-d-glueonic acid. A treatment of 6% solution of 2-deoxy-d-glucose with commercial glucose oxidase preparation caused also a complete dehydrogenation.     

Palynological study of a holocene peat and a miocene coal deposit from NW Borneo

The development of a Holocene peat near Marudi (Sarawak) and a Miocene coal near Berakas (Brunei) is studied pollen-analytically and stratigraphically by comparison with present-day swamp vegetation along the NW Bornean coast. A number of 76 pollen and spore types were recognized and are briefly diagnosed.The Marudi profile starts on mangrove sediment and shows a succession which is closely comparable to the present-day zonation of the large raised bog of which it forms part.The Berakas profile starts on fluviatile sediments and remains in the mixed swamp forest stage. This small deposit has remained close to the coast and peat growth is terminated by marine transgression.The floristic composition of the mixed swamp forest stage in both bogs, separated in age by approximately seven million years is closely comparable. Minor differences which exist can be attributed to differences in size, ecologic conditions and accessibility from mainland reservoirs.Only on spore type, Stenochlaena areolaris has become extinct in Borneo.     

The ultrastructure of topical cutaneous calcinosis

Summary Mineralized plaques, which develop at the site of repeated subcutaneous injections of 100 g KMnO₄/0.2 H₂O in rats, were investigated by electron microscopy. The newly formed, delineated, white plaque tissue at the injection site consisted of numerous, mostly unaltered fibroblasts and collagen fibers, without participation of inflammatory cells. Some signs of cell injury were found in the center of the lesions. Numerous, irregularly distributed, small, mineralized foci were seen near the fibroblasts. These were formed by aggregation of small needle-like units (50 Å in diameter and 0.05–2.0 m long). These needle-shaped units were found either in vesicular, cell derived structures, considered to be shed cell processes or cell fragments, or on collagen fibers. Intramitochondrial deposits of such needle-like units were seen frequently. Fusion of smaller mineralized foci to larger plaques occured and then needle-shaped units were seen at the periphery of the electron-dense lesions. Hypotheses concerning the mechanism of experimental cutaneous calcinosis (soft tissue mineralization) are discussed and related to the findings of this study. Probable intracellular crystal deposition and mineralization in cell-derived structures were shown for the first time in topical cutaneous calcinosis.     

A Highlights from MBoC Selection: Fast and parallel nanoscale three-dimensional tracking of heterogeneous mammalian chromatin dynamics

Chromatin organization and dynamics are critical for gene regulation. In this work we present a methodology for fast and parallel three-dimensional (3D) tracking of multiple chromosomal loci of choice over many thousands of frames on various timescales. We achieved this by developing and combining fluorogenic and replenishable nanobody arrays, engineered point spread functions, and light sheet illumination. The result is gentle live-cell 3D tracking with excellent spatiotemporal resolution throughout the mammalian cell nucleus. Correction for both sample drift and nuclear translation facilitated accurate long-term tracking of the chromatin dynamics. We demonstrate tracking both of fast dynamics (50 Hz) and over timescales extending to several hours, and we find both large heterogeneity between cells and apparent anisotropy in the dynamics in the axial direction. We further quantify the effect of inhibiting actin polymerization on the dynamics and find an overall increase in both the apparent diffusion coefficient D* and anomalous diffusion exponent α and a transition to more-isotropic dynamics in 3D after such treatment. We think that in the future our methodology will allow researchers to obtain a better fundamental understanding of chromatin dynamics and how it is altered during disease progression and after perturbations of cellular function.