A Notch/Delta-dependent relay mechanism establishes anterior-posterior polarity in Drosophila

The anterior-posterior axis of Drosophila becomes polarized early in oogenesis, when the oocyte moves to the posterior of the germline cyst because it preferentially adheres to posterior follicle cells. The source of this asymmetry is unclear, however, since anterior and posterior follicle cells are equivalent until midoogenesis, when Gurken signaling from the oocyte induces posterior fate. Here, we show that asymmetry arises because each cyst polarizes the next cyst through a series of posterior to anterior inductions. Delta signaling from the older cyst induces the anterior polar follicle cells, the anterior polar cells signal through the JAK/STAT pathway to induce the formation of the stalk between adjacent cysts, and the stalk polarizes the younger anterior cyst by inducing the shape change and preferential adhesion that position the oocyte at the posterior. The anterior-posterior axis is therefore established by a relay mechanism, which propagates polarity from one cyst to the next.     

Regularity in irregular echinoids

 Using the mathematical concept of eutactic star, we prove that it is possible to define a morphospace for irregular echinoids by using a single parameter. In particular, we have found an extraordinary geometric property in the flower-like patterns of the five ambulacral petals of these animals. This property is fulfilled with great accuracy for a large collection of fossil specimens and provides new insights in the study of the viable skeletal designs of extinct and/or living organisms. Received: 19 June 2001 / Revised version: 2 October 2001 / Published online: 14 March 2002     

A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi,which interacts with the structural flagellar protein paraflagellar rod

Trypanosoma cruzi adenylyl cyclases are encoded by a large polymorphic gene family. Although several genes have been identified in this parasite, little is known about the properties and regulation of these enzymes. Here we report the cloning and characterization of TczAC, a novel member of T. cruzi adenylyl cyclase family. The TczAC gene is expressed in all of the parasite life forms and encodes a 1,313-amino acid protein that can complement a Saccharomyces cerevisiae mutant deficient in adenylyl cyclase activity. The recombinant enzyme expressed in yeasts is constitutively active, has a low affinity for ATP (K(m) = 406 microm), and requires a divalent cation for catalysis. TczAC is inhibited by Zn(2+) and the P-site inhibitor 2'-deoxyadenosine 3'-monophosphate, suggesting some level of conservation in the catalytic mechanism with mammalian adenylyl cyclases. It shows a dose-dependent stimulation by Ca(2+) which can be reversed by high concentrations of phenothiazinic calmodulin inhibitors. However, bovine calmodulin fails to stimulate the enzyme. Using a yeast two-hybrid screen it was found that TczAC interacts through its catalytic domain with the paraflagellar rod protein, a component of the flagellar structure. Furthermore, we demonstrate that TczAC can dimerize through the same domain. These results provide novel evidence of the possible localization and regulation of this protein.     

Role of adipocyte differentiation-related protein in surfactant phospholipid synthesis by type II cells

Adipocyte differentiation-related protein (ADrP) is an intrinsic lipid storage droplet protein that is highly expressed in lung. ADrP localizes to lipid storage droplets within lipofibroblasts, pulmonary cells characterized by high triacylglycerol, which is a precursor for surfactant phospholipid synthesis by alveolar type II epithelial (EPII) cells. The developmental pattern of ADrP mRNA and protein expression in lung tissue parallels triacylglycerol accumulation in rat lung. ADrP mRNA levels are relatively high in isolated lipofibroblasts, accounting for the high ADrP expression in lung. Isolated EPII cells, which do not store neutral lipids but derive them from lipofibroblasts, have low levels of ADrP mRNA expression. ADrP is found around lipid droplets in cultured lipofibroblasts, but not in EPII cells isolated from developing rat lung. After coculture with lipofibroblasts, EPII cells acquired ADrP, which associates with lipid droplets. Furthermore, (3)H-labeled triolein in isolated ADrP-coated lipid droplets is a tenfold better substrate for surfactant phospholipid synthesis by cultured EPII cells than (3)H-labeled synthetic triolein alone. Antibodies to ADrP block transfer of neutral lipid. These data suggest a role for ADrP in this novel mechanism for the transfer of lipid between lipofibroblasts and EPII cells.     

Leptin mediates the parathyroid hormone-related protein paracrine stimulation of fetal lung maturation

Developing rat lung lipofibroblasts express leptin beginning on embryonic day (E) 17, increasing 7- to 10-fold by E20. Leptin and its receptor are expressed mutually exclusively by fetal lung fibroblasts and type II cells, suggesting a paracrine signaling "loop." This hypothesized mechanism is supported by the following experimental data: 1) leptin stimulates the de novo synthesis of surfactant phospholipid by both fetal rat type II cells (400% x 100 ng(-1) x ml(-1) x 24 h(-1)) and adult human airway epithelial cells (85% x 100 ng(-1) x 24 h(-1)); 2) leptin is secreted by lipofibroblasts in amounts that stimulate type II cell surfactant phospholipid synthesis in vitro; 3) epithelial cell secretions such as parathyroid hormone-related protein (PTHrP), PGE(2), and dexamethasone stimulate leptin expression by fetal rat lung fibroblasts; 4) PTHrP or leptin stimulate the de novo synthesis of surfactant phospholipid (2- to 2.5-fold/24 h) and the expression of surfactant protein B (SP-B; >25-fold/24 h) by fetal rat lung explants, an effect that is blocked by a leptin antibody; and 5) a PTHrP receptor antagonist inhibits the expression of leptin mRNA by explants but does not inhibit leptin stimulation of surfactant phospholipid or SP-B expression, indicating that PTHrP paracrine stimulation of type II cell maturation requires leptin expression by lipofibroblasts. This is the first demonstration of a paracrine loop that functionally cooperates to induce alveolar acinar lung development.     

Cadmium removal by living cells of the marine microalga Tetraselmis suecica

Cadmium removal by living cells of the marine microalga Tetraselmis suecica was tested in cultures exposed to different cadmium concentrations (0.6, 3, 6, 15, 30 and 45 mg/l). The EC50 for growth was 7.9 mg Cd/l after six days of exposure. The cadmium removed was proportional to the concentration of this metal in the medium and it was dependent on the time of exposure; cultures with higher cadmium concentration removed a higher amount of this metal. In cultures exposed to 0.6 mg/l, T suecica cells removed 98.1% of added cadmium with 0.392 x 10(-6) microg Cd/cell, whereas in cultures with 45 mg/l only 7.7% was removed with 16.052 x 10(-6) microg Cd/cell. The highest amount of cadmium removed per liter of culture was observed in cultures exposed to 6 mg/l, with 3.577 mg/l of cadmium. After six days of incubation, the higher proportion of cadmium was bioaccumulated intracellularly in all cultures except in 45 mg/l cultures, the percentage of intracellular cadmium being always more than 50%. The highest percentage of bioadsorbed cadmium (60.1%) was found in cells of cultures with the highest cadmium concentration (45 mg/l). Furthermore, a relation between intracellular cadmium and the concentration of sulfhydryl groups was observed.     

PCNA protein associates to Cdk-A type protein kinases in germinating maize

In higher eukaryotes, the proliferating cell nuclear antigen (PCNA) can be found associated to Cyclin D and Cdk4/6, the kinase complex responsible for cell cycle commitment in response to growth and mitogenic signals. During maize germination, PCNA can be found in protein complexes between 131 and 163 kDa. The sizes of PCNA protein complexes seem to change during germination, so that by the time the S phase starts, a complex of 100 kDa (likely the homotrimeric ring) is the predominant one. PCNA complexes during early germination contain (any of) two PSTAIRE-containing protein kinases of 32 and 36 kDa that readily phosphorylate both histone H1 and maize retinoblastoma-related (RBR) proteins. Kinase activity in PCNA complexes is markedly inhibited by roscovitine and olomoucine, two known Cdk inhibitors. The protein p13^Suc1 only pulls down the 36 kDa PSTAIRE protein. Kinase activity in PCNA immunoprecipitates is maximal during early germination, before the onset of the S-phase, whereas kinase activity associated to p13^Suc1 reaches a peak later, after the onset of the S-phase. We discuss the physiological repercussions of these findings.     

Population growth rate of the depredating Podisus nigrispinus (Heteroptera: Pentatomidae) and of the Tuta absoluta (Leptoptera: gelechiidae) in wintering place

The fertility life table of Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) preying either on Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) caterpillars or on alternative prey Tenebrio molitor L. (Coleoptera: Tenebrionidae) under greenhouse conditions (30 +/- 5 degrees C, 61 +/- 23% RH) were studied. The life table was also determined for the pest T. absoluta under the same conditions. The net reproductive rate (Ro) and the intrinsic rate of natural increase (rm) were higher 14.13 and 46.32 times for predators fed on T. molitor prey, however, the generation time (T) was similar between prey. The pest T. absoluta showed Ro and rm higher 2.15 and 32.10 times than those achieved for predators fed on this pest. However, females fed on a suitable prey T. molitor showed higher Ro and rm than those yielded for the pest. The survival curves were similar for P. nigrispinus females fed on both prey and classified as being type II by Weibull analysis. The results suggest that P. nigrispinus is able to maintain its population preying only on T. absoluta caterpillars; however, the life table parameters determined individually for both showed that the pest produces more generations per year and faster population natural growth than the predator.     

NMR triple-quantum filtered relaxation analysis of O-water in insulin solutions: an insight into the aggregation of insulin and the properties of its bound water

Transverse triple-quantum filtered NMR spectroscopy (TTQF) of ¹⁷O-water was used to study the properties of water in insulin solutions at different Zn²⁺ concentrations and pH values. It was established that strongly bound water molecules are already present in Zn-free insulin. On the assumption that the effective correlation time of a strongly bound water molecule, τ_sb, is 10 ns, the apparent number of strongly bound water molecules was 3 to 4 per insulin monomer. Addition of Zn²⁺ equivalent to 2 g-atoms per hexamer did not produce substantial increases in the overall ¹⁷O-water TTQF signal intensity and apparent fraction of bound water. The dramatic enhancement of the TTQF signals observed for samples with a Zn²⁺/hexamer ratio greater than 2:1 could be attributed to the increase in correlation time of the strongly bound water, due to the formation of higher-order oligomers of the protein.