Molecular cloning of a pair of human pepsinogen A genes which differ by a Glu→Lys mutation in the activation peptide

Summary Three human cosmid clones containing pepsinogen A (PGA) encoding sequences were isolated from a genomic bank derived from a single individual. One cosmid contains two PGA genes in tandem in a head-to-tail orientation, while the other two cosmids each contain a single PGA gene. The three cosmids were characterized by restriction mapping and sequence analysis (exons 1 and 2 and flanking regions). As judged from these data, three of the four PGA genes isolated appear to be nearly identical, but one of the tandem genes is clearly different from the other genes. The first exon of all four genes codes for the same amino acid sequence. However, in the second exon of one of the tandem genes we found a nucleotide substitution giving rise to a GluLys substitution of the 43rd amino acid residue of the activation peptide, leading to a charge difference of the corresponding isozymogens. The presence of two distinct PGA genes in the isolated gene pair conclusively proves the multigene structure of the PGA system. These genes might be responsible for at least part of the electrophoretic polymorphism at the protein level.     

The influence of crabs on litter processing in high intertidal mangrove forests in tropical Australia

Summary Measurements of litter fall and litter removal by crabs, in conjunction with estimates of litter decay by microbes and tidal export of litter from three high-intertidal mangrove forests were made during a year-long study in tropical northeastern Australia. In forests dominated by Ceriops tagal and Bruguiera exaristata, litter standing stocks remained low on the forest floor (mean 6 g·m^-2), although litter fall was high; 822 and 1022 g·m^-2·y^-1, respectively. Sesarmid crabs removed 580 (Ceriops) and 803 (Bruguiera) g·m^-2·y^-1, or 71 and 79%, of the total annual litter fall from the forest floor. Relative to the rate of litter removal by crabs, microbial turnover of whole, unshredded litter was insignificant, accounting for <1% of annual litter fall. Export of litter by tides was estimated to remove 194 (Ceriops) and 252 (Bruguiera) g·m^-2·y^-1 or 24 and 25% of annual litter fall. In a forest dominated by Avicenniamarina, in which an ocypodid crab was more abundant than sesarmids, litter standing stocks were higher (mean 84 g·m^-2) and crabs removed less litter; 173 g·m^-2·y^-1 or 33% of the annual litter fall of 519 g·m^-2·y^-1. Microbial turnover of intact litter was more important in the Avicennia forest (168 g·m^-2·y^-1 or 32% of annual litter fall), and tides exported 107 g·m^-2·y^-1 or 21% of litter production. In areas where sesarmid crabs were absent or rare in Ceriops forests, there were significantly higher standing stocks of litter and slower rates of leaf removal. Taking into account the probable assimilation efficiencies of sesarmid crabs feeding on mangrove leaves, we estimate that in Ceriops and Bruguiera forests leaf processing by crabs turns litter over at >75 times the rate of microbial decay alone, thus facilitating the high sediment bacterial productivity in these forests. The importance of litter processing by crabs increases with height in the intertidal in tropical Australia, in contrast to New World mangrove forests, where the reverse is true.Contribution No. 445 from the Australian Institute of Marine Science     

Auxin Physiology of the Tomato Mutant diageotropica

The tomato (Lycopersicon esculentum, Mill.) mutant diageotropica (dgt) exhibits biochemical, physiological, and morphological abnormalities that suggest the mutation may have affected a primary site of auxin perception or action. We have compared two aspects of the auxin physiology of dgt and wild-type (VFN8) seedlings: auxin transport and cellular growth parameters. The rates of basipetal indole-3-acetic acid (IAA) polar transport are identical in hypocotyl sections of the two genotypes, but dgt sections have a slightly greater capacity for IAA transport. 2,3,5-Triiodobenzoic acid and ethylene reduce transport in both mutant and wild-type sections. The kinetics of auxin uptake into VFN8 and dgt sections are nearly identical. These results make it unlikely that an altered IAA efflux carrier or IAA uptake symport are responsible for the pleiotropic effects resulting from the dgt mutation. The lack of auxin-induced cell elongation in dgt plants is not due to insufficient turgor, as the osmotic potential of dgt cell sap is less (more negative) than that of VFN8. An auxin-induced increase in wall extensibility, as measured by the Instron technique, only occurs in the VFN8 plants. These data suggest dgt hypocotyls suffer a defect in the sequence of events culminating in auxin-induced cell wall loosening.     

Postcranial adaptation and evolution in lorisidae

With the exception of leaping, lorises and galagos move in many similar ways although frequencies and styles differ. This peculiar locomotor distinction in two closely related subfamilies has profoundly altered their respective postcranial anatomies from their common ancestor. A comparison of postcranial adaptation in extant forms shows that lorises and galagos differ somewhat in forelimb mobility, but are more fundamentally disparate in hindlimb adaptation. Inferences concerning locomotor adaptation in the lorisid fossil record indicate a more generalized locomotor pattern which is more like that of extant cheirogaleids than either living galagos or lorises. Thus, vertical clinging and leaping in galagines and the slow-climbing and suspensory movements of lorisines appear to be evolutionarily recent innovations from a more generalized locomotor past.     

High extracellular concentrations of potassium and rubidium modulate the synaptic transmission in the frog labyrinth

High extracellular K or Rb levels (20 mM) produce an increase in the resting EPSP and spike frequencies recorded intra cellularly from single fibres of the posterior nerve in the isolated frog labyrinth. The afferent discharge facilitation proved to be inversely related to the fibre's initial resting activity. The K effect is systematically larger than the Rb effect. High sensitive and scarcely sensitive units may be identified with respect to K and Rb action. The present findings suggest that, according to previous models of hair cell functioning, the K and Rb effects are mediated by a raise in intracellular Ca concentration which sustains an increased transmitter release at the cyto-neural junction.     

Effects of salinity on chloride cells in the euryhaline cyprinodontid fish Rivulus marmoratus

Summary The ultrastructure and density of chloride cells in the gill, opercular epithelium, and opercular skin of the euryhaline self-fertilizing fish Rivulus marmoratus (Cyprinodontidae) were studied with electron and fluorescence microscopy. R. marmoratus raised from birth in 1, 50, 100, and 200% seawater were compared. Chloride cells from fish raised in each of the four salinities exhibited an invaginated pit structure at the apical crypt. Multicellular complexes were present in the 1% seawater group and in those fish raised in higher salinities where elaborate interdigitations were seen between cells. Chloride cells from gills of fish raised in 200% seawater had a significantly higher percentage of their cytoplasmic volume composed of mitochondria than did those from fish raised in 1% seawater (69.9% vs 37.4%). The opercular skin and opercular epithelium had the same density of chloride cells (4.2×10⁴-4.5×10⁴ chloride cells/cm²), and this number did not vary significantly with increased salinity. The opercular skin thus appears far more responsive to environmental salinity than the opercular epithelium. Chloride cells from the opercular epithelium of fish raised in 200% seawater were found to be 39% larger than those from fish raised in 1% seawater, whereas the chloride cells from the opercular skin of the 200% seawater group were 107% larger than those from the 1% seawater group.     

Prevention of the normal expansion of maternal plasma volume: a model for chronic fetal hypoxaemia

The effects of inadequate expansion of maternal blood volume on uterine blood flow, fetal oxygen levels and vasoactive mediators during the third trimester were studied in 8 pregnant sheep. Results were compared to those obtained during 15 normal pregnancies. Prevention of the normal (20 ml/day) increase in maternal plasma volume was achieved by repeated haemorrhage and injections of furosemide. These treatments also reduced the rise in blood flow to the pregnant uterine horn that normally occurs during this period of gestation: at term flow was only 508 +/- 61 (SEM) compared to 838 +/- 83 ml/min in the control group (P greater than 0.01). This reduction in uterine blood flow caused a gradual fall in fetal PaO2, and rise in fetal levels of plasma renin activity, vasopressin, catecholamines and angiotensin II without change in pHa or base excess. Four to 5 days prior to delivery, the difference from control in PaO2 was -3.9 +/- 0.5 mmHg, plasma renin activity +2.9 +/- 1.7 ng/ml.h, vasopressin +4.2 +/- 1.1 pg/ml, catecholamines +957 +/- 145.3 pg/ml and angiotensin II +243 +/- 108.2 pg/ml. Furthermore, the fall in PaO2 and rise in vasoactive mediators that normally occur 3-5 days prior to the onset of labour was either absent (PaO2 and plasma renin activity) or blunted. Thus when expansion of blood volume during pregnancy is inadequate, blood flow to the uterus is adversely affected. This leads to various degrees of chronic fetal hypoxaemia and stimulation of vasoactive mediator systems. However, the normal stimulation of vasoactive mediator systems that occurs 3-5 days before delivery appears to be blunted. Experimental prevention of blood volume expansion during pregnancy produces an excellent model for the study of chronic mild fetal hypoxaemia.     

Effect of a hyperlipidic diet on lipid composition, fluidity, and (Na+-K+)ATPase activity of rat erythrocyte membranes

Feeding rats a hyperlipidic diet in which animals were offered daily a variety of high-energy food resulted in a significant increase of serum free fatty acids and a decrease of phospholipids with respect to controls. On the contrary, there were no significant differences in erythrocyte membrane total lipid composition between the two groups. Erythrocyte membranes showed a significant decrease in saturated fatty acid content and a significant increase in (n-6) polyunsaturated fatty acid content; (n-3) polyunsaturated fatty acids significantly decreased. Membrane fluidity, investigated by fluorescence polarization of diphenylhexatriene, significantly increased in the erythrocyte membranes of the experimental group. These results seem compatible with the decreased saturated/unsaturated fatty acid ratio. A significant decrease of (Na+-K+)ATPase activity occurred in erythrocyte membranes of the experimental group rats with respect to the controls.     

2',3'-dideoxycytidine permeation of the human erythrocyte membrane

The mechanism by which 2,3'-dideoxycytidine, an inhibitor of HIV-I infectivity, permeates the cell membrane was investigated. The influx of ddCyd into human erythrocytes was nonconcentrative. The initial velocity of both ddCyd influx and efflux was, in contrast to compounds that permeate the cell membrane via the nucleoside transporter, a linear function of nucleoside concentration in the 1 microM to 10 mM range and relatively insensitive to temperature. Furthermore, potent inhibitors of nucleoside transporter and other nucleosides were found to inhibit ddCyd influx only partially or not at all suggesting that ddCyd permeates the human erythrocyte membrane predominantly by nonfacilitated diffusion. This unusual characteristic seems to be due to the lack of 3'-hydroxyl moiety of ddCyd which appears to be an important determinant for the nucleoside carrier specificity rather than to lipid solubility itself. As far as permeation of the cell membrane is concerned ddCyd shares these properties with 2',3'-dideoxythymidine and 3'-azido-3'-deoxythymidine.