Human proteins IEF 58 and 57a are associated with the Golgi apparatus

A mouse monoclonal antibody (mAB 22-II-D8B) raised against lysed transformed human amnion cells (AMA) has been characterized. The mAB decorated the Golgi apparatus in growing and quiescent cultured monolayer cells (fibroblasts and epithelial cells) of various species as determined by double immunofluorescence labeling and colocalization with galactosyltransferase antibodies. It reacted with the acidic human proteins IEF 58 (Mr = 29,000) and 57a, respectively (Mr = 30,000) (HeLa protein catalogue number; [(1982) Clin. Chem. 28, 766]), Golgi staining was also observed in BS-C-1 cells microinjected with mAB 22-II-D8B suggesting that the epitopes recognized by the antibody are most likely located on the cytoplasmic face of the membranes. The precise localization of the antigens to the various cisternae of the Golgi apparatus could not be demonstrated by immunogold cytochemistry on ultrathin cryosections due to either weak reactivity of the antibody or low concentration of the antigens. Immunofluorescence staining with mAB 22-II-D8B of lymphoid human Molt-4 cells and some human tissues failed to reveal any significant staining even though these expressed high levels of both IEF 58 and 57a. These results are taken to imply that the epitopes recognized by mAB 22-II-D8B may be masked in some cell types.     

THE FINE STRUCTURE OF CHONDROCOCCUS COLUMNARIS : I. Structure and Formation of Mesosomes

Cells of Chondrococcus columnaris were sectioned and examined in the electron microscope after fixation by two different methods. After fixation with osmium tetroxide alone, the surface layers of the cells consisted of a plasma membrane, a dense layer (mucopeptide layer), and an outer unit membrane. The outer membrane appeared distorted and was widely separated from the rest of the cell. The intracytoplasmic membranes (mesosomes) appeared as convoluted tubules packaged up within the cytoplasm by a unit membrane. The unit membrane surrounding the tubules was continuous with the plasma membrane. When the cells were fixed with glutaraldehyde prior to fixation with osmium tetroxide, the outer membrane was not distorted and separated from the rest of the cell, structural elements (peripheral fibrils) were seen situated between the outer membrane and dense layer, and the mesosomes appeared as highly organized structures produced by the invagination and proliferation of the plasma membrane. The mesosomes were made up of a series of compound membranes bounded by unit membranes. The compound membranes were formed by the union of two unit membranes along their cytoplasmic surfaces.     

THE FINE STRUCTURE OF CHONDROCOCCUS COLUMNARIS : II. Structure and Formation of Rhapidosomes

When cells of C. columnaris were broken open, treated with PTA, and examined in the electron microscope, tubular structures (rhapidosomes) were present in the preparations. The rhapidosomes are approximately 300 A in diameter. Their length varies from about 500 to about 15,000 A. An axial hole which runs the length of the rhapidosomes appears to widen and narrow with a regular periodicity. End-on views of short segments of rhapidosomes revealed the presence of subunits around their outside peripheries. The results of studies of lysed cells and of sectioned cells indicate that the rhapidosomes are produced during the disintegration of cells. It seems likely that the compound membranes of the mesosomes break down to give rise to the tubular structures. The mesosomal origin of rhapidosomes is postulated only for the rhapidosomes of C. columnaris, since the origin of rhapidosomes from other organisms was not investigated during this study. The rhapidosomes of C. columnaris may be unrelated to those of S. grandis, S. myxococcoides, A. violaceum, and Sorangium 495, since there was a difference in the details of fine structure between rhapidosomes from C. columnaris and those found in the other four organisms.     

THE FINE STRUCTURE OF TWO UNUSUAL STALKED BACTERIA

Two strains of bacteria that produce slender appendages (pseudostalks) from their lateral surfaces were studied using the electron microscope. The pseudostalks were shown to be extensions of the cytoplasm and peripheral membranes of the cell proper. Both strains of bacteria produce holdfasts at the poles of the cells by the means of which attachment can take place. The pseudostalks are not involved in the attachment of cells. No specialized intracytoplasmic structures are present at the point of juncture of pseudostalk and cell. A discussion of the possible functions of the pseudostalks, based on the electron microscope findings, is presented.     

Estimation of the effect of photoinhibition on the carbon gain in leaves of a willow canopy

The occurrence of photoinhibition of photosynthesis in leaves of a willow canopy was examined by measuring the chlorophyll-a fluorescence ratio of F _V/F _M (F_M is the maximum fluorescence level of the induction curve, and F_V is the variable fluorescence, F _V=F _M–F ₀, where F₀ is the minimal fluorescence). The majority of the leaves situated on the upper parts of peripheral shoots showed an afternoon inhibition of this ratio on clear days. This was the consequence of both a decrease in F _M and a rise in F _O. In the same leaves the diurnal variation in intercepted photosynthetic photon flux density (PPFD) was monitored using leaf-mounted sensors. Using the multivariate method, partial least squares in latent variables, it is shown that the dose of PPFD, integrated and linearly weighted over the last 6-h period, best predicts photoinhibition. Photoinhibition occurred even among leaves that did not intercept PPFDs above 1000 mol·m^–2·s^–1. Exposure of leaves to a standard photoinhibitory treatment demonstrated that the depression in the F _V/F _M ratio was paralleled by an equal depression in the maximal quantum yield of CO₂ uptake and a nearly equal depression in the rate of bending (convexity) of the light-response curve of CO₂ uptake. As a result, the rate of net photosynthesis is depressed over the whole natural range of PPFD. By simulating the daily course in the rate of net photosynthesis, it is estimated that in the order of one-tenth of the potential carbon gain of peripheral willow shoots is lost on clear days as a result of photoinhibition. This applies to conditions of optimal temperatures. Photoinhibition is even more pronounced at air temperatures below 23° C, as judged from measurements of the F_V/F_M ratio on clear days: the afternoon inhibition of this ratio increased in a curvilinear manner from 15% to 25% with a temperature decrease from 23° to 14° C.Abbreviations and Symbols F_O minimum fluorescence - F_V variable fluorescence - F_M maximum fluorescence - PLS partial least squares in latent variables - PPFD photosynthetic photon flux density - VPD water vapour-pressure deficit This study was supported by the Swedish Natural Science Research Council. We are indebted to Dr. Jerry Leverenz (Department of Plant Physiology, University of Umeå, Sweden) for guidance with the modelling of the photosynthesis data.     

The importance of macrophyte bed size for cladoceran composition and horizontal migration in a shallow lake

Cladoceran composition and diel horizontal migration were studiedin 2, 10 and 25 m diameter macrophyte exclosures establishedin the littoral zone of shallow Lake Stigsholm, Denmark. Theexclosures were protected from waterfowl grazing, but open tofish. The macrophyte community cornprized Potwnogeton pectinatus,Potamogeton pusillus and Callitriche hemaphroditica. Cladoceranswere sampled randomly every third hour inside and outside themacrophyte exclosures during a 24 h period. In the 2m exclosure,the pelagic species Ceriodaphnia spp. and Bosmina spp. dominatedduring the day, mean density being as high as 3430 indiv. l^–1During the night, density decreased to 10–20% of the daytimedensity thus indicating diel horizontal migration. In the 10and 25 m exciosures, the daytime mean density of Ceriodaphniaspp. was 865 and 202 indiv. l^–1, respectively, and didnot decrease at night. In contrast to the pelagic species, thedensity of macrophyte-associated species tended to be higherin the 10 and 25 m exclosure than in the 2 m exclosure. In thedaytime, Eurycercus lamellatus density in the 2, 10 and 25 rnmacrophyte exclosures was 7, 28 and 16 indiv. l^–1 respectively,while that of Simocephalus vetulus was 11, 171 and 92 mdiv.l^–1, respectively. There was no thy-night difference inthe density of macrophyte-associated species. We conclude thatcladoceran community composition varies with macrophyte bedsize, and that the edge zone between the bed and open wateris an important daytime refuge for potentially migrating pelagiccladocerans.     

High Numbers of Prosthecate Bacteria in Pulp Mill Waste Aeration Lagoons

Prosthecate bacteria comprised 0.6 to 10.5% of the bacterial community in samples from 11 pulp mill waste aeration lagoons. Because of their distinct morphology, the genera Ancalomicrobium, Caulobacter, Prosthecobacter, Prosthecomicrobium, Stella, and Hyphomicrobium or Hyphomonas could be identified and enumerated by direct microscopic examination. Monthly samples from one lagoon showed that several genera varied from undetectable to predominant among the appendaged organisms. Temperature (season), type of wood pulped, and pulping process did not significantly affect the density of prosthecate bacteria.     

Plasma and Tissue Levels of Trimethoprim in The Rainbow Trout,Salmo Gairdneri,After Absorption From Fresh and Salt Water

This is a comparative study of uptake of trimethoprim from 1) fresh water, 2) salt water after 7 days of adaption and 3) salt water without previous adaptation. The rate and extent of absorption were found to vary significantly. The salt water adapted group reached a plasma concentration of approx. 1 µg/ml after 10 h, the unadapted salt water group after 24 to 48 h and the fresh water group did not reach this concentration. The results are discussed in relation to the non-ionic diffusion theory and to the alterations taking place in euryhaline species of fish during adaptation to salt water.     

Structure and toxicity of pure ricinus agglutinin

Highly purified ricinus agglutinin was found to inhibit protein synthesis in HeLa cells. This effect could be prevented by the addition of the specific antiricinus agglutinin serum, whereas specific anti-ricin serum did not protect the cells, demonstrating that the toxic effect of ricinus agglutinin is not due to contamination with ricin.After reduction of ricinus agglutinin with 2-mercaptoethanol in the presence of 0.5 M galactose the constituent peptide chains were separated by chromatography on a DE-52 column. The B′-chain passed through the column, whereas the A′-chain bound and was eluted with a salt gradient. The B′-chain was further purified by chromatography on a CM-52 column.The shortest chain, the A′-chain, was found to inhibit cell-free protein synthesis, whereas the B′-chain did not have this ability. On the other hand, the B′-chain was able to induce agglutination of erythrocytes when tested together with anti-ricinus agglutinin serum indicating that the B′-chains bind to the cells.Ouchterlony immunodiffusion tests with crude anti-ricin and anti-ricinus agglutinin sera revealed that the two constituent chains of ricinus agglutinin are immunologically partial identical and that they also show reaction of partial identity with both chains of the toxic lectin ricin.The data indicate that a similar structure-function relationship exists in ricinus agglutinin as in ricin. The reason for the much lower toxicity of ricinus agglutinin than of ricin in living animals is discussed.