Personal Light Dosimetry in Permanent Night and Day Workers

Light exposure was measured in six day and six night watches (working 12-hour shifts five days in a row) during 48 h on work days and 48 h on days off using a photocell with a sensitivity corresponding to photopic vision. The photocell was mounted on a frame of spectacles, thus measuring in viewing direction. Light exposure was low both in night and day watches; however, in night watches exposures were significantly lower: On work days, night watches spent a mean of 13 min above 1,500 lx, day watches 52 min; on days off, night watches spent 3 min above 1,500 lx but day watches 89 min. Unexpectedly, night watches had no higher exposure during days off. We suspect that this is due to a light avoidance tendency in permanent night workers. High negative correlations between the acrophases of subjective state (e.g., alertness and mood) and light exposure in night watches indicate that bright light would probably increase desynchroniza-tion between subjective state, sleep, and activity.     

Characterization of a cell cycle-dependent nuclear autoantigen

Analysis of reactivity to nuclear antigens in autoimmune sera revealed a serum that produced a previously undescribed cell cycle-dependent immunofluorescence staining pattern. By indirect immunofluorescence using HEp-2 cells as substrate, the serum generated a speckled and nucleolar pleomorphic staining pattern. This characteristic immunofluorescence pattern was detected in different cell lines from various species indicating that the antigen was highly conserved. This serum immunoprecipitated a 85 kDa protein using an extract from [³⁵S]-labeled HeLa cells. Indirect immunofluorescence of proliferating mouse 3T3 cells displayed the characteristic pleomorphic staining which was not observed in serum-starved cells. Resting human and mouse peripheral blood lymphocytes were negative in immunofluorescence while mitogen-stimulated lymphocytes were positive. Germinal centers of mice two weeks after immunization with 2-phenyl-oxazolone showed speckled immunofluorescence staining in the dark zones whereas unimmunized mice were completely negative. Cell synchronization experiments showed a characteristic sequence of locations of the antigen during the cell cycle. In G₁, cells were completely negative. In late G₁, G₁/S and S phase, speckles were visible. In early G₂, speckles were visible, and later in G₂, the nucleoli were positive. During mitosis chromosomes were stained. Further characterization of this antibody specificity and cloning of cDNA are in progress.     

Cell-free in vitro reduction of carboxylates to aldehydes: With crude enzyme preparations to a key pharmaceutical building block

The scarcity of practical methods for aldehyde synthesis in chemistry necessitates the development of mild, selective procedures. Carboxylic acid reductases catalyze aldehyde formation from stable carboxylic acid precursors in an aqueous solution. Carboxylic acid reductases were employed to catalyze aldehyde formation in a cell-free system with activation energy and reducing equivalents provided through auxiliary proteins for ATP and NADPH recycling. In situ product removal was used to suppress over-reduction due to background enzyme activities, and an N-protected 4-formyl-piperidine pharma synthon was prepared in 61% isolated yield. This is the first report of preparative aldehyde synthesis with carboxylic acid reductases employing crude, commercially available enzyme preparations.     

Das Corno-Quercetum des Mátra-Gebirges

Zusammenfassung Die zum Aceri-Quercion-Verband gehörenden kontinentalen und kalkholden Corno-Quercetum Wälder sind in dem nordöstlichen Ungarischen Mittelgebirge so im Mátra-Gebirge auf Andesit, auf Kalk- und vulkanischem Grundgestein verbreitet. Einige Bestände der Corno-Quercetum Wälder stehen im Kontakt mit dem auf Lößboden (Tschernosjom-brauner Waldboden) vorhandenen Aceri tatarici-Quercetum des Flachlandes (Randgebiet der Ungarischen Tiefebene).Die artenreichen Bestände sind durch den verhältnismäßig hohen Anteil der Aceri-Quercion Charakterarten und der sog. Waldsteppenarten gut gekennzeichnet.In der floristischen Zusammensetzung des Corno-Quercetum spielen außer den obererwähnten Arten die Elemente von Quercetea pubescenti-petraeae und Querco-Fagetea die Hauptrolle.Auf Grund der Charakterisierung des Standortes der Corno-Quercetum Wälder durch die T-, W- und R-Indikatorwerte der Arten nach Zólyomi (1963, 1964) ist das Corno-Quercetum als eine relativ basischen, mäßig trockenen Standort kennzeichnende, an submediterranen wärmeliebenden Arten reiche Gesellschaft kontinentale Gepräges zu betrachten.Auf Andesit-Grundgestein entstandene Böden (Erubasboden, brauner Waldboden, Parabraunerde) des Corno-Quercetum sind reicher an adsorbiertem Ca, und das ermöglicht das Erscheinen der Bestände und so auch der Arten, die einen gewissen Kalkgehalt und eine neutrale, oder schwach basische Reaktion des Bodens beanspruchen.

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Summary The author describes the Corno-Quercetum oak forests from the Mátra-Mts. in Hungary.Floristic composition indicates requirements for chalk and nitrogen. The author also includes a number of ecological data (temperature, oil-composition, humidity) measured in these forests.

     

A RAPD study of the Sarcostemma group of Cynanchum (Apocynaceae-Asclepiadoideae-Asclepiadeae)

One hundred and thirty-seven accessions of Cynanchum viminale and its relatives, formerly known as Sarcostemma, were studied using randomly amplified polymorphic DNA (RAPD). A fingerprinting technique was used because sequencing had failed to differentiate between morphologically separable groups. Chromosome counts were conducted to establish the ploidy level of the accessions. The banding patterns resulting from RAPD analysis were evaluated with Canonical Analysis of Principal Coordinates, Permanova and neighbour-joining. A strong geographic component was found in the structure of the group. Taxa considered species or subspecies based on morphology often formed coherent groups. The data are interpreted to reflect at least two cycles of diversification: the first one from Madagascar and the second one most likely from the East African–Arabian region, reaching Madagascar again. In mainland Africa, polyploidisation has occurred several times.     

Characterization of an S-layer glycoprotein produced in the course of S-layer variation of<Emphasis Type="Italic"> Bacillus stearothermophilu</Emphasis>s ATCC 12980 and sequencing and cloning of the<Emphasis Type="Italic"> sbsD</Emphasis> gene encoding the protein moiety

The cell surface of Bacillus stearothermophilus ATCC 12980 is completely covered by an oblique lattice which consists of the S-layer protein SbsC. On SDS-polyacrylamide gels, the mature S-layer protein migrates as a single band with an apparent molecular mass of 122 kDa. During cultivation of B. stearothermophilus ATCC 12980 at 67 degrees C instead of 55 degrees C, a variant developed that had a secondary cell wall polymer identical to that of the wild-type strain, but it carried an S-layer glycoprotein that could be separated on SDS-polyacrylamide gels into four bands with apparent molecular masses of 92, 118, 150 and 175 kDa. After deglycosylation, only a single protein band with a molecular mass of 92 kDa remained. The complete nucleotide sequence encoding the protein moiety of this S-layer glycoprotein, termed SbsD, was established by PCR and inverse PCR. The sbsD gene of 2,709 bp is predicted to encode a protein of 96.2 kDa with a 30-amino-acid signal peptide. Within the 807 bp encoding the signal peptide and the N-terminal sequence (amino acids 31-269), different nucleotides for sbsD and sbsC were observed in 46 positions, but 70% of these mutations were silent, thus leading to a level of identity of 95% for the N-terminal parts. The level of identity of the remaining parts of SbsD and SbsC was below 10%, indicating that the lysine-, tyrosine- and arginine-rich N-terminal region in combination with a distinct type of secondary cell wall polymer remained conserved upon S-layer variation. The sbsD sequence encoding the mature S-layer protein cloned into the pET28a vector led to stable expression in Escherichia coli HMS174(DE3). This is the first example demonstrating that S-layer variation leads to the synthesis of an S-layer glycoprotein.