Resonance Raman characterization of the 7-ns photoproduct of (carbonmonoxy)hemoglobin: implications for hemoglobin dynamics

Resonance Raman spectra are reported for deoxyhemoglobin (deoxyHb) and the (carbonmonoxy)hemoglobin (HbCO) photoproduct Hb by use of 7-ns YAG laser pulses at wavelengths of 416 and 532 nm, where enhancement is observed for totally symmetric and nontotally symmetric modes, respectively. The frequencies of the porphyrin skeletal modes v10, v2, v19, v11, and v3 have been determined to be 1602, 1559, 1553, 1542, and 1466 cm-1 in Hb. These frequencies are 2-3 cm-1 lower than the corresponding frequencies for deoxyHb. The v19 and v11 frequencies are at the expected values for a Ct-N distance of 2.057 A, the known core size for a 6-coordinate high-spin FeII-porphyrin complex. The remaining frequencies, however, deviate from the core size correlations for these modes in the same direction as do those of deoxyHb, suggesting that the porphyrin ring is domed in both species. Thus, the heme structure is similar for deoxyHb and Hb but is slightly expanded in the latter. The expanded heme in Hb implies a restraint on the full out-of-plane displacement of the Fe atom, by an estimated approximately 0.1 A relative to deoxyHb. This could result from a residual interaction with the CO molecule if the latter remains held by the protein against the Fe atom, in a high-spin 6-coordinate complex. The available spectroscopic evidence suggests that such a complex may be stabilized at 4 K but is unlikely to persist at room temperature beyond the electronic relaxation (0.35 ps) of the electronically excited heme.(ABSTRACT TRUNCATED AT 250 WORDS)     

Binding of purified Escherichia coli O75X adhesin to frozen sections of human kidney

Abstract Binding characteristics of the purified Escherichia coli O75X adhesin in frozen sections of human kidney were determined, using antibodies raised against the purified antigen and rhodamine-conjugated second antibodies. To identify the adhesin-binding nephron domains, the same tissue sections were double stained with fluorescein isothiocynate-conjugated nephron site-specific lectins. The results revealed that, at the tubular pole, the O75X adhesin bound selectively to the basement membrane of proximal and distal tubules and, with a slightly lower efficiency, of collecting ducts. In the glomerulus, the O75X adhesin bound only to the parietal epithelial cells (Bowman's capsule). The purified O75X adhesin bound also to exfoliated epithelial cells in human urine. Our results suggest that the O75X adhesin may contribute to the uropathogenicity of E. coli by binding the bacteria to tissue structures in the human urinary tract.     

Control of phosphorus loading and flushing as restoration methods for Lake Veluwe,The Netherlands

As a result of high nutrient loading Lake Veluwe suffered from an almost permanent bloom of the blue-green algaOscillatoria agardhii Gomont. In 1979, the phosphorus loading of the lake was reduced from approx. 3 to 1 g P.m^–2.a^–1. Moreover, since then the lake has been flushed during winter periods with water low in phosphorus. This measure aimed primarily at interrupting the continuous algal bloom. The results of these measures show a sharp decline of total-phosphorus values from 0.40–0.60 mg P.l^–1 (before 1980) to 0.10–0.20 mg P.l^–1 (after 1980). Summer values for chlorophylla dropped from 200–400 mg.m^–3 to 50–150 mg.m^–3.The increase in transparency of the lake water was relatively small, from summer values of 15–25 cm before the implementation of the measures to 25–45 cm afterwards. The disappointing transparency values may be explained by the decreasing chlorophylla and phosphorus content of the algae per unit biovolume. Blue-green algae are gradually loosing ground. In the summer of 1985 green algae and diatoms dominated the phytoplankton for the first time since almost 20 years. To achieve the ultimate water quality objectives (transparency values of more than 100 cm in summer), the phosphorus loading has to be reduced further.     

Studies on thyroid cell surface glycoproteins: Isolation of plasma membranes and characterization of carbohydrate units

Plasma membranes were isolated from calf thyroid microsomes and further resolved into two subfractions by sucrose density gradient centrifugations. The lighter and major membrane fraction was obtained in a yield of 10 mg/100 g of thyroid and was enriched 38-fold with respect to 5′-nucleotidase activity compared to the homogenate. It differed from the denser plasma membrane fraction in containing greater amounts of phospholipid and cholesterol but had a similar total carbohydrate content (16 mg/100 mg protein) and monosaccharide composition. The membranes were found to retain most (80%) of their carbohydrate after delipidation. The major protein-bound sugars present in the lighter membrane fraction expressed as micromoles per 100 mg of peptide were: galactose 24, mannose 17, fucose 3, glucosamine 23, galactosamine 4, and sialic acid 9. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate of the lipid-free membranes revealed at least 18 protein bands and 3 periodic acid-Schiffreactive glycoprotein components. Incubation of the delipidated membranes with Pronase resulted in the solubilization of 95% of the saccharide portion which upon filtration through Bio-Gel P-6 and P-10 columns yielded several glycopeptide fractions. While some of the carbohydrate was found in glycopeptides which appeared to contain the well-known complex and polymannose asparagine-bound oligosaccharides, as well as small O-glycosidically linked units, approximately half was recovered in high molecular weight components which contained galactose and glucosamine as their principal sugar constituents, and which were similar in composition to glycopeptides recently isolated (T. Krusius, J. Finne, and H. Rauvala, 1978, Eur. J. Biochem.92, 289–300) from human erythrocyte membranes.     

Biological evaluation of the systematic validity of the African Argas (Persicargas) Arboreus and the Asian-Australian A. (P.) Robertsi (Ixodoidea: Argasidae)

Khalil G. M., Hoogstraal H. and Oliver J. H. Jr. 1980. Biological evaluation of the systematic validity of the African Argas (Persicargas) arboreus and the Asian-Australian A.(P.)robertsi (Ixodoidea: Argasidae). International journal for Parasitology10: 253–259. The closely related A. (P.)arboreus (Ethiopian Faunal Region) and A. (P.) robertsi (Oriental and Australian Faunal Regions) interbreed readily and produce partially fertile progeny. Intrahybrid crosses are much less productive than P₁ homogamic crosses. Results of backcrosses indicate that the progeny of the robertsi ♂ × arboreus ♀ cross are less fertile than the progeny of the reciprocal cross. The fertility of F₁ hybrid progeny is lower than that of the P₁ pure species as expressed in lower female fecundity, egg hatch, and/or viability of immature stages. These 2 species probably originated from a common ancestor and geographic isolation caused genetic incompatibility.     

Calcium oxalate crystals in plants

Calcium (Ca) oxalate crystals occur in many plant species and in most organs and tissues. They generally form within cells although extracellular crystals have been reported. The crystal cells or idioblasts display ultrastructural modifications which are related to crystal precipitation. Crystal formation is usually associated with membranes, chambers, or inclusions found within the cell vacuole(s). Tubules, modified plastids and enlarged nuclei also have been reported in crystal idioblasts. The Ca oxalate crystals consist of either the monohydrate whewellite form, or the dihydrate weddellite form. A number of techniques exist for the identification of calcium oxalate. X-ray diffraction, Raman microprobe analysis and infrared spectroscopy are the most accurate. Many plant crystals assumed to be Ca oxalate have never been positively identified as such. In some instances, crystals have been classified as whewellite or weddellite solely on the basis of their shape. Certain evidence indicates that crystal shape may be independent of hydration form of Ca oxalate and that the vacuole crystal chamber membranes may act to mold crystal shape; however, the actual mechanism controlling shape is unknown. Oxalic acid is formed via several major pathways. In plants, glycolate can be converted to oxalic acid. The oxidation occurs in two steps with glyoxylic acid as an intermediate and glycolic acid oxidase as the enzyme. Glyoxylic acid may be derived from enzymatic cleavage of isocitric acid. Oxaloacetate also can be split to form oxalate and acetate. Another significant precursor of oxalate in plants is L-ascorbic acid. The intermediate steps in the conversion of L-ascorbic acid to oxalate are not well defined. Oxalic acid formation in animals occurs by similar pathways and Ca oxalate crystals may be produced under certain conditions. Various functions have been attributed to plant crystal idioblasts and crystals. There is evidence that oxalate synthesis is related to ionic balance. Plant crystals thus may be a manifestation of an effort to maintain an ionic equilibrium. In many plants oxalate is metabolized very slowly or not at all and is considered to be an end product of metabolism. Plant crystal idioblasts may function as a means of removing the oxalate which may otherwise accumulate in toxic quantities. Idioblast formation is dependent on the availability of both Ca and oxalate. Under Ca stress conditions, however, crystals may be reabsorbed indicating a storage function for the idioblasts for Ca. In addition, it has been suggested that the crystals serve purely as structural supports or as a protective device against foraging animals. The purpose of this review is to present an overview of plant crystal idioblasts and Ca oxalate crystals and to include the most recent literature.     

Distribution of diatom species on an estuarine mud flat and experimental analysis of the selective effect of stress

The distribution patterns of seven dominant diatom species on an estuarine mud flat transect were related to salinity gradients and organic pollution. The temporal distribution of species can be explained partly by the seasonal variation in irradiance and temperature, and partly by the large discharges of organically polluted fresh water during the late autumn. The selective effects of stress factors, such as low or high salinity, high concentrations of ammonia and free sulphide, and high temperatures were studied by measuring: (1) the tolerance of natural diatom populations isolated from the sediment; (2) the tolerance and capacities of unialgal cultures: and (3) the effects of the stress factors on the species composition of the populations in sediment samples incubated in the laboratory. A low salinity (1‰), but not a high salinity (20‰), markedly altered the species composition of natural diatom populations kept in the laboratory. Navicula salinarum Grunow and N. cf. cryptocephala Kützing outcompete other species only at a salinity of 1‰, which is below the optimum salinity for growth of these species. High concentrations (2–4 mM) of ammonia inhibited the photosynthesis of N. phyllepta Kützing and N. flanatica Grunow and decreased the cell numbers of these species in mixed populations kept in the laboratory. N. salinarum and Gyrosigma fasciola (Ehr) Cleve were relatively ammonia-tolerant and reached their highest abundance during periods of large discharges of organically polluted water. The populations on the mud-flat stations that had black, sulphide-containing layers 1 mm below the surface of the sediment, were dominated by the relatively sulphide-tolerant Navicula salinarum and N. pygmaea Kützing.High values of irradiance and temperature were a selective factor explaining the absence of N. flanatica in summer. Uptake of organic substrates and the inhibitory effect of high population densities are discussed in regard to their possible modifying influence on the distribution of diatom species on the mud flat.     

Electron microscopic study of the origin and formation of Reissner's fiber in the subcommissural organ of Cebus apella (Primates,Platyrrhini)

Summary Two kinds of secretion are formed in cells of the subcommissural organ (SCO) of Cebus apella. The light secretion is found in saccules originating from the endoplasmic reticulum. This secretion is stored in the peripheral portion of the cells and is not involved in formation of Reissner's fiber (RF). In close association with the Golgi complex, electron-dense granules are developed, containing a finely granular substance. These granules accumulate beneath the apical plasmalemma of the cell. Their content is discharged into the third ventricle, where it occurs in the form of a thin layer of secretion. This material appears to constitute the RF at the level of the entrance to the mesencephalic aqueduct.