Summer dynamics of the deep chlorophyll maximum in Lake Tahoe

Vertical profiles of chlorophyll and phytoplankton biomass weremeasured in Lake Tahoe from July 1976 through April 1977. Adeep chlorophyll maximum (DCM) persisted during summer and earlyautumn (July—October) near 100 m, well below the mixedlayer and at the upper surface of the nitracline. The DCM coincidedwith the phytoplankton biomass maximum as determined from cellcounts. In addition, the composition of the phytoplankton assemblagewas highly differentiated with respect to depth. Cyclotellastelligera was the predominant species in the mixed layer whilethe major species in the DCM layer included C. ocellata andseveral green ultraplanktonic species. In situ cell growth playsa substantial role in maintaining the DCM, but sinking of cellsfrom shallower depths and zooplankton grazing above the DCMmay contribute to the maintenance of the DCM. Calculations supportthe interpretation that the summer DCM persists at the boundarybetween an upper, nutrient-limited phytoplankton assemblageand a deeper, light-limited assemblage.     

Kinetic studies of four enzyme forms of beta-N-acetylhexosaminidase from embryonic chicken brain

The separation of four different hexosaminidase forms from embryonic chicken brain (16-day-old) has been performed by ion-exchange chromatography. Two different DEAE-cellulose columns have been used: a first one at pH 7.2 and a second one at pH 6.0. Km and Vmax values were estimated from the Lineweaver-Burk or Dixon plots and ki from the Dixon plots, using N-acetyl-D-glucosamine or N-acetyl-D-galactosamine as inhibitors. In both cases we found a kind of competitive inhibition in which Lineweaver-Burk and Dixon plots curve downwards.     

Isolation, characterization and phospholipid composition of lamellar bodies and subcellular fractions from dog lung

1. Lamellar body fractions from dog lung can be separated by a procedure based on differential centrifugation before ultracentrifugation onto a discontinuous sucrose gradient. This fraction yields about 1% of total protein from the homogenate. 2. The different fractions obtained in the isolation were assayed for the measurement of four subcellular marker enzymes: beta-N-acetylglucosaminidase, acid phosphatase, 5'-nucleotidase and succinate dehydrogenase. 3. Lamellar bodies were not contaminated by mitochondria (0.7 succinate dehydrogenase relative specific activity), whereas high specific hydrolase activities were found (beta-N-acetylglucosaminidase and 5'-nucleotidase were enriched 1.8- and 2.8-fold, respectively). 4. The chemical criterion was established by measuring the specific components of lamellar bodies. The lamellar bodies have the highest phospholipid/protein ratio (0.35); cholesterol/protein ratio (0.15) and the highest phosphatidylglycerol percentages (7.9%). 5. The phospholipid composition of lamellar bodies is distributed among phosphatidylcholine (64.5%), phosphatidylethanolamine (11%), phosphatidylglycerol (7.9%), sphingomyelin (4%), phosphatidylserine and phosphatidylinositol (3%), respectively. The remainder were considered as trace amounts (less than 1%).     

The organization of repetitive sequences in the albumin and alpha-fetoprotein gene loci in the rat

Summary The distribution of middle repetitive sequences in the genic and extragenic regions of the rat albumin and -fetoprotein genes was analyzed. Their presence was determined by probing Southern blots of restriction fragments of albumin and -fetoprotein genomic subclones with ³²P-labeled total rat DNA. Repetitive sequences were detected in both genes. They were classified as weak, moderate and intense hybridizing elements according to the intensity of hybridization. Weak repetitive sequences were characterized as dG·dT repeats by using ³²P-labeled poly-(dG·dT)(dC·dA) oligomer probe. They occurred in 5 and 3 extragenic regions of the two genes and in introns 4 and 5 of the albumin gene. The moderate repetitive sequence present in intron 6 of the albumin gene was identified as the rat SINES element, 4D12. The intense repetitive sequence, localized in the 3 non-coding region of the albumin gene, corresponded to the terminal segment of a rat high repeat long interspersed DNA family, L1Rn. 4D12 and L1Rn sequences were also scattered throughout the -fetoprotein locus as moderate and intense repetitive elements, respectively, but their distribution was different from that of the albumin genomic region. These results indicate that repetitive sequences invaded the two loci in a non-conservative manner.     

An improved purification procedure and properties of casein kinase II from brain

A simple and short purification procedure applicable to casein kinase II has been developed, for fully characterizing the enzyme from calf cerebral cortex cytosol. The procedure consists of four chromatographic steps: DEAE-cellulose, phosphocellulose, phosvitin-Sepharose and ATP-agarose which yields 87% pure casein kinase II. The purified enzyme shows three major bands with apparent molecular masses of 42, 38, and 27 kDa by polyacrylamide gel electrophoresis in sodium dodecyl sulfate and is self-autophosphorylated on its 27 kDa polypeptide. The enzyme shows all the characteristics described for casein kinase II from other sources: it is independent of cyclic nucleotides, calcium/phospholipids, and double-stranded poly(I).poly(C); it can utilize both ATP and GTP as phosphoryl donors and can phosphorylate both casein and phosvitin but not histone. The kinetic studies establish that theK _m for ATP is 12.5 M and 25.1 M when using phosvitin and casein respectively as phosphoryl acceptors. TheK _m for phosvitin is 0.91 mg/ml and for casein 1.43 mg/ml, while theV _max is 315 nmol/min/per mg protein and 479 nmol/min/per mg protein for phosvitin and casein respectively. The activity of the kinase is highly stimulated by KCl or NaCl, and almost completely inhibited by heparin concentrations of 1 g/ml (92%). This inhibition is reduced to only 33% in the presence of optimal KCl concentrations (150 mM). Spermine stimulates enzyme activity, whilst hemin produces a slight inhibition.     

Externalization of aminophospholipids in platelet liposome bilayers by supplementation with sphingomyelin

Negatively charged phospholipids are mainly located on the inner leaflet of platelet liposomes. Using a specific reagent for amino groups, trinitrobenzenesulfanilic acid, we have shown that in large artificial vesicles prepared with total lipids from sheep platelets supplemented with increased amounts of exogenous sphingomyelin, an alteration in the amino-phospholipid topology occurs, with a progressive appearance of these lipid species -in particular phosphatidylserine- in the outer membrane bilayer.