Cloning and expression of two human p70 S6 kinase polypeptides differing only at their amino termini.

Two classes of human cDNA encoding the insulin/mitogen-activated p70 S6 kinase have been isolated; the two classes differ only in the 5' region, such that the longer polypeptide (p70 S6 kinase alpha I; calculated Mr 58,946) consists of 525 amino acids, of which the last 502 residues are identical in sequence to the entire polypeptides encoded by the second cDNA (p70 S6 kinase alpha II; calculated Mr 56,153). Both p70 S6 kinase polypeptides predicted by these cDNAs are present in p70 S6 kinase purified from rat liver, and each is thus expressed in vivo. Moreover, both polypeptides are expressed from a single mRNA transcribed from the (longer) p70 S6 kinase alpha I cDNA through the utilization of different translational start sites. Although the two p70 S6 kinase polypeptides differ by only 23 amino acid residues, the slightly longer alpha I polypeptide exhibits anomalously slow mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), migrating at an apparent Mr of 90,000 probably because of the presence of six consecutive Arg residues immediately following the initiator methionine. Transient expression of p70 alpha I and alpha II S6 kinase cDNA in COS cells results in a 2.5- to 4-fold increase in overall S6 kinase activity. Upon immunoblotting, the recombinant p70 polypeptides appear as a closely spaced ladder of four to five bands between 65 and 70 kDa (alpha II) and 85 and 90 kDa (alpha I). Transfection with the alpha II cDNA yields only the smaller set of bands, while transfection with the alpha I cDNA generates both sets of bands. Mutation of Met-24 in the alpha I cDNA to Leu or Thr suppresses synthesis of the alpha II polypeptides. Only the p70 alpha I and alpha II polypeptides of slowest mobility on SDS-PAGE comigrate with the 70- and 90-kDa proteins observed in purified rat liver S6 kinase. Moreover, it is the recombinant p70 polypeptides of slowest mobility that coelute with S6 kinase activity on anion-exchange chromatography. The slower mobility and higher enzymatic activity of these p70 proteins is due to Ser/Thr phosphorylation, inasmuch as treatment with phosphatase 2A inactivates kinase activity and increases the mobility of the bands on SDS-PAGE in an okadaic acid-sensitive manner. Thus, the recombinant p70 S6 kinase undergoes multiple phosphorylation and partial activation in COS cells. Acquisition of S6 protein kinase catalytic function, however, is apparently restricted to the most extensively phosphorylated recombinant polypeptides.     

Transmission electron microscopical studies of the site of attachment of Ancylostoma ceylanicum to the small bowel mucosa of the dog

The host-parasite interaction at the site of attachment of the adult hookworm, Ancylostoma ceylanicum, to the small bowel mucosa of the dog has been examined by transmission electron microscopy. The lamina propria around the heads of the worms showed intense infiltration with neutrophilic and eosinophilic leucocytes, and plasma cells were also prominent. Erythrocytes were extravasated within the lamina propria while the portions of mucosa engulfed into the buccal cavities of the worms were necrotic and included aggregates of collagen fibres. Within the lumina of the worms were erythrocytes in varying stages of lysis together with other necrotic debris. The mucous membrane adjacent to the heads of the worms was ulcerated while more distant enterocytes were small, distorted and displayed various degrees of microvillar loss. Many mucosal blood vessels had platelet aggregates within their lumina but no fibrin deposition was observed. Vessels were often seen in the bases of the ulcers and were prone to rupture. Erythrocytes and leucocytes were seen in the dog intestinal lumen in close proximity to the bodies of the worms.     

Two‐step vegetation response to enhanced precipitation in Northeast Brazil during Heinrich event 1

High resolution palynological and geochemical data of sediment core GeoB 3910‐2 (located offshore Northeast Brazil) spanning the period between 19 600 and 14 500 calibrated year bp (19.6–14.5 ka) show a land‐cover change in the catchment area of local rivers in two steps related to changes in precipitation associated with Heinrich Event 1 (H1 stadial). At the end of the last glacial maximum, the landscape in semi‐arid Northeast Brazil was dominated by a very dry type of caatinga vegetation, mainly composed of grasslands with some herbs and shrubs. After 18 ka, considerably more humid conditions are suggested by changes in the vegetation and by C_org and C/N data indicative of fluvial erosion. The caatinga became wetter and along lakes and rivers, sedges and gallery forest expanded. The most humid period was recorded between 16.5 and 15 ka, when humid gallery (and floodplain) forest and even small patches of mountainous Atlantic rain forest occurred together with dry forest, the latter being considered as a rather lush type of caatinga vegetation. During this humid phase erosion decreased as less lithogenic material and more organic terrestrial material were deposited on the continental slope of northern Brazil. After 15 ka arid conditions returned. During the humid second phase of the H1 stadial, a rich variety of landscapes existed in Northeast Brazil and during the drier periods small pockets of forest could probably survive in favorable spots, which would have increased the resilience of the forest to climate change.     

Studies on Chloroplast Degradation in vivo: III. THE EFFECT OF MAGNESIUM ON ELECTRON TRANSPORT AND PHOSPHORYLATIQN IN PLASTIDS ISOLATED FROM YOUNG AND AGEING COTYLEDONS

The magnesium dependence of NADP-reduction, oxygen evolutionand concomitant ATP-formation was studied as a function of theage of the Cucurbita cotyledons from which the chloroplastswere isolated. Whereas NADP-reduction under alkaline assay conditionsalways exhibits a stimulation upon addition of Mg⁺⁺, measurementsin an acidic medium show an age-dependent reversal to inhibition.The data for oxygen evolution are more complex in nature, dueto participation of consecutive reactions. The Mg⁺⁺ concentrationfor optimal ATP formation decreases with the age of the cotyledonarymaterial. The results are discussed from the viewpoint of apossible control of essential photosynthetic functions in vivoby magnesium.     

THE 'TRANSITION PROBABILITY MODEL' AND THE REGULATION OF PROLIFERATION OF HUMAN DIPLOID CELL CULTURES DURING AGING

Genealogies derived from time-lapse cinemicrophotographic studies of aging human diploid cell cultures were analysed in terms of the ‘transition probability’ model. It was found that the distribution of intermitotic times obtained from middle passage cells deviated only slightly from that predicted by the model. In contrast, the plot for late passage cultures did not fit the predicted pattern and appeared to be composed of multiple curves. These changes are discussed in reference to cellular senescence as expressed by normal human diploid cells in vitro.     

Rate of growth in length of Bangladeshi infants as a function of attained length.

We examine variation in the rate of growth in length of breast-feeding infants from rural Bangladesh. These data were collected between November 1985 and February 1986 from two rural sites. Eighty-eight infants, ranging from birth to 4 months of age at the start of the study and their mothers were measured monthly for 4 months. Length increased linearly with age over this 4-month period (infants' average bias-adjusted R2 = 0.90). The relationship between infant rate of growth in length and attained length was analyzed by two different methods: Oldham's (1962) method of regressing rate of growth on mean length and Blomqvist's (1977) method of regressing rate of growth on estimated initial length. The methods gave similar results. The rate of growth was negatively associated with mean infant length over the 4-month period (p less than 0.001); that is, shorter infants grew at a faster rate than longer infants. For every centimeter shorter the infant was, the rate of growth was 0.1 cm/mo faster on average; the effect was greater among males than among females. The average rate of growth was greater for males than for females and greater in financially solvent households and varied by site. Infant growth rate was slower among older infants than among younger infants, as expected. However, after adjusting for mean infant length, age was no longer significantly associated with infant growth rate, although mean infant length remained highly significant. Forty-one percent of the variation in infant rate of growth in length was explained by mean infant length, sex, sex by length interaction, household financial solvency, and site.