The influence of cholecystokinin, vasoactive intestinal peptide and secretin on pancreatic and biliary secretion in laying hens

White Leghorn hens, 14-29 weeks old, were surgically fitted with cannulas for collecting pancreatic and biliary secretions, and a jugular cannula for continuous infusion of either cholecystokinin (CCK), vasoactive intestinal peptide (VIP), or secretin. As compared to secretory levels during saline infusion, CCK significantly stimulated biliary flow and biliverdin concentration in bile; VIP significantly depressed biliverdin concentration but enhanced bicarbonate secretion in both pancreatic and biliary secretions, and also increased total pancreatic flow. Secretin depressed biliary flow and increased pancreatic bicarbonate release. The principal hormonal regulator of biliary secretion appears to be CCK, and that of pancreatic secretion to be VIP.     

Sucrose synthase and invertase in isolated vascular bundles

Vascular bundles were isolated from grapefruit (Citrus paradisi Macf.) during periods of rapid sucrose translocation into fruit. Invertase and sucrose synthase activities were assayed in these strands and compared with immediately adjacent tissues (inner most peel and segment epidermis) and phloem-free juice sacs during four growing seasons. Although sucrose synthase was present in sink cells, the significantly greater activity in vascular strands (per unit fresh weight and protein) indicated that the role of this enzyme in translocation may include a vascular function in addition to its proposed involvement in metabolism of importing cells.     

Standardized isolation of protoplasts

Isolation of leaf mesophyll protoplasts from tobacco (Nicotiana tabacum) is facilitated using a specially designed digestion chamber. The chamber's airtight seals and various ports reduce the complexity, time and contamination risks which may be associated with standard isolations. The polished glass viewing window allows continuous monitoring of protoplast release, thus facilitating more precise determinations of the optimal digestion time. In concert with a simplified centrifugation step, the protoplast isolation procedure is greatly standardized.     

Human keratinocytes maintain reversible anti-apoptotic defenses in vivo and in vitro

Human keratinocytes proliferate and differentiate in an epidermal environment where induction of apoptosis can be triggered by ultraviolet radiation (UVR), activated lymphocytes and cytokines. The purpose of this study was to determine whether keratinocytes were susceptible to apoptosis induced by ionophore, ultra-violet radiation, cytokines or crosslinking of CD95 (Fas/APO-1). In normal human skin exposed to two minimal erythema doses of ultraviolet radiation, suprabasal cells were the first keratinocytes to demonstrate apoptotic nuclei, and by 48 h apoptotic cells were identified throughout the mid to upper epidermis. However, most keratinocytes resisted apoptosis and UVR-induced apoptosis was not observed in basal cells, or in the most differentiated epidermis. Human keratinocytes and keratinocyte cell lines cultured in vitro developed maximal apoptosis 48 h after radiation. Human keratinocytes cultured in full growth factor supplements were resistant to UVR-induced apoptosis compared to keratinocyte cell lines or to a lymphoid cell line (HL60) susceptible to apoptosis. Keratinocyte cell lines were completely resistant to apoptosis induced by interferon-, interferon-, IL-2, IL-6, TNF-, IL-1Ra, and GM-CSF. A subset of the cells in cultures of keratinocytes and transformed keratinocyte cell lines died by apoptosis in response to anti-Fas, IL-1 and TNF- plus IFN- and ionophore. Second passage freshly isolated human keratinocytes were much more resistant to apoptosis induced by ionophore, anti-Fas and cytokines than were transformed keratinocyte cell lines. Calcium shift to induce differentiation in second-passage keratinocyte cultures made keratino-cytes even more resistant to UVR-induced apoptosis. This parallels the lack of UVR-induced apoptosis observed in the most differentiated keratinocytes in irradiated human skin. Both keratinocytes and kerati-nocyte cell lines express rather low levels of the anti-apoptotic proteins bcl-2 and bcl-x compared to other apoptosis-resistant cell types. The differences between keratinocytes and keratinocyte cell lines in suscepti-bility to apoptosis are not explained by difference in expression of bcl-2 or bcl-x. Finally, withdrawal of growth factors from keratinocytes decreased cell survival following UVR and increased the induction of apoptosis. Inhibition of protein synthesis with cyclo-heximide also made keratinocytes more susceptible to UVR-induced apoptosis, indicating that anti-apop-totic defences in cultured keratinocytes are dependent on active protein synthesis. These experiments show that the strong keratinocyte defences against apoptosis are stratified within the epidermis, and can be altered by differentiation and growth factor withdrawal.     

Two-dimensional motion of DNA bands during 120° pulsed-field gel electrophoresis. I. Effect of molecular weight

The instantaneous position and velocity of bands of linear, double-stranded DNA were measured during 120° pulsed-field electrophoresis in 1% agarose gels, using a video micrometer capable of simultaneous measurements in two dimensions. When the direction of the field was switched, the band initially retraced the last portion of its path during the preceding pulse. The distance the band moved backward increased with DNA length: 48.5 kb (kilobase pair) DNA moved backward only 0.2 μm, but 1110 kb DNA moved backward 24 μm, before setting off in a positive direction. The velocity of the DNA band was particularly rapid during the backward movement: the magnitude of the velocity spike increased with M, reaching 2.4 μm/s for 1110 kb DNN, which was about 5 times the steady-state velocity. The velocity in the y direction, perpendicular to the mean drift direction, allowed an even larger transient spike, which also increased with M. Simulation of the dynamics of long DNA chins undergoing gel electrophoresis by a dynamic Monte Carlo method gave instantaneous xy position and velocity in excellent agreement with experiment. The simulation included extensional motions of the DNA within the tube of interconnected agarose pores as well as the possibility of loops (hernias) that escape laterally from the tube. © 1995 John Wiley & Sons, Inc.     

Molecular phylogenetic evidence that the phylum Haplosporidia has an alveolate ancestry

The phylogenetic position of the phylum Haplosporidia among other protists was investigated with the complete 16S-like rRNA gene sequences from two species in the phylum: Haplosporidium nelsoni, a parasite of oysters, and Minchinia teredinis, a parasite of shipworms. Because the lack of obvious morphological homologies with other protists hampered decisions regarding taxonomic composition for sequence alignment and phylogenetic analysis, the complete sequences for these two haplosporidians were directed as search queries to the blast/ncbi.nlm.nih.gov electronic mail server. The results of this heuristic similarity search provided a basis for constructing a preliminary higher-taxonomic-level analysis comparing the haplosporidians with species from the slime molds, fungi, algae, amoebae, ciliates, dinoflagellates, and apicomplexans. Maximum parsimony yielded equivocal results, whereas transversionally weighted parsimony suggested an affinity with the alveolates (i.e., the ciliates, dinoflagellates, and apicomplexans). Multiple alignment of the two haplosporidian sequences against 17 taxa in a secondary analysis focusing on the alveolates and subsequent parsimony analysis placed the phylum Haplosporidia as a monophyletic group within the Alveolata and as a taxon of equal rank with the other three alveolate phyla. The precise placement within the Alveolata was sensitive to weighting.      

Effects of Glyphosate on Metabolism of Phenolic Compounds VIII. Comparison of the Effects of Aminooxyacetate and Glyphosate

Aminooxyacetate (AOA), an in vitro inhibitor of phenylalanineammonia-lyase (PAL) and of some transaminases, was tested forcomparison with glyphosate's [N-(phosphonomethyl)glycine] effectson plant growth, PAL activity, and accumulation of hydroxyphenoliccompounds of three-day-old, dark-grown soybean [Glycine max(L.) Merr.] seedlings. Root-fed AOA (50 µM) and glyphosate(0.5 mM) caused similar decreases in growth rate and in accumulationof hydroxyphenolics, anthocyanin and chlorophyll. Together,these compounds were neither antagonistic nor synergistic inaffecting these parameters. AOA caused decreases in extractablePAL activity while increasing aromatic amino acid pools—theopposite of glyphosate's effects. In the light, the effect ofglyphosate on PAL and aromatic amino acids predominated overthose of AOA when the chemicals were given together. The effectsof AOA and glyphosate on most free amino acid levels were similar.In those cases in which the effects differed, glyphosate's effectpredominated over that of AOA when the chemicals were giventogether. The similarities and interactions of AOA and glyphosatein their effects on free amino acid profiles indicate that glyphosatesignificantly interferes with non-aromatic as well as aromaticamino acid synthesis. (Received April 6, 1982; Accepted June 28, 1982)     

Tentoxin effects onSorghum: The role of polyphenol oxidase

Summary In an ultrastructural and cytochemical study of tentoxin-treatedSorghum bicolor (L.) Moench, both bundle sheath and mesophyll plastids were severely affected, Plastids from chlorotic leaf areas lacked most internal membranes yet had plastid ribosomes and large fibrillar areas of plastid DNA. In recovered areas (mottled yellow and green), cells were found that had plastids of near-normal ultrastructure as well as the severely affected plastid-types found in chlorotic leaf areas. Polyphenol oxidase (PPO) cytochemistry of these mottled leaf areas indicated that all recovered mesophyll plastids had PPO whereas all the abnormal mesophyll plastids showed no activity. Because bundle sheath plastids ofSorghum have no PPO activity at any developmental stage, yet are affected by tentoxin, PPO cannot be uniquely affected by this toxin. We suggest that tentoxin may affect the transport of cytosolic proteins into the plastid.     

Role of the testa in preventing cellular rupture during imbibition of legume seeds

Studies with the seeds of soybean, navy bean, pea, and peanut were made to determine the extent of leakage of intracellular enzymes during imbition. Embryos with intact testae from all four species were found to leak detectable activities of either intracellular enzymes of the cytosol (glucose-6-phosphate dehydrogenase) or enzymes found in both the cytosol and organelles (malate dehydrogenase, glutamate dehydrogenase, glutamate oxaloacetate transaminase, and NADP-isocitrate dehydrogenase) after 6 hours imbition at 25 C. Pea and peanut embryos with testae leaked considerably lower levels of activity for these enzymes than did those of soybean and bean. Leakage of mitochondrial marker enzymes (fumarase, cytochrome c oxidase, and adenylate kinase) was not detected from embryos with testae, suggesting that a differential diffusion of intracellular components out of cells occurred. Soybean and bean embryos without testae leaked high, and proportionally (per cent dry seed basis) similar, levels of all cytosol, cytosol-organelle, and mitochondrial marker enzymes and protein during imbibition, indicating that cell membranes were not differential to leakage and that they had ruptured. Pea and peanut embryos without testae leaked detectable activities of all cytosol and cytosol-organelle enzymes, although fumarase was the only detectable mitochondrial marker enzyme leaked, suggesting that some degree of differential leakage may have occurred in these species. The outermost layers of embryo cells of seeds without testae of all four species absorbed and sequestered the nonpermeating pigment Evan's blue after 5 to 15 minutes imbibition, indicating that membranes had ruptured. This occurred to a much lesser extent in seeds with intact testae. Both soybean and bean embryos without testae were observed to disintegrate during imbibition, whereas those of pea and peanut did not. These data indicate that seeds of certain legumes are susceptible to cellular rupture during imbibition when seed coats are damaged or missing.