Intrahepatic bile ducts transport water in response to absorbed glucose

The physiological relevance of theabsorption of glucose from bile by cholangiocytes remains unclear. Theaim of this study was to test the hypothesis that absorbed glucosedrives aquaporin (AQP)-mediated water transport by biliary epitheliaand is thus involved in ductal bile formation. Glucose absorption andwater transport by biliary epithelia were studied in vitro bymicroperfusing intrahepatic bile duct units (IBDUs) isolated from ratliver. In a separate set of in vivo experiments, bile flow andabsorption of biliary glucose were measured after intraportal infusionof D-glucose or phlorizin. IBDUs absorbedD-glucose in a dose- and phlorizin-dependent manner with anabsorption maximum of 92.8 ± 6.2 pmol · min¹ · mm¹.Absorption of D-glucose by microperfused IBDUs resulted inan increase of water absorption (J_v = 310nl · min¹ · mm¹,P_f = 40 × 10³ cm/sec).Glucose-driven water absorption by IBDUs was inhibited byHgCl₂, suggesting that water passively followsabsorbed D-glucose mainly transcellularly viamercury-sensitive AQPs. In vivo studies showed that as the amount ofabsorbed biliary glucose increased after intraportal infusion ofD-glucose, bile flow decreased. In contrast, as theabsorption of biliary glucose decreased after phlorizin, bile flowincreased. Results support the hypothesis that the physiologicalsignificance of the absorption of biliary glucose by cholangiocytes islikely related to regulation of ductal bile formation.

     

Nongastric H-K-ATPase in rodent prostate: lobe-specific expression and apical localization

The molecular basis of active iontransport in secretory glands such as the prostate is not wellcharacterized. Rat nongastric H-K-ATPase is expressed at highlevels in distal colon surface cell apical membranes and thus isreferred to as "colonic." Here we show that the ATPase is expressedin rodent prostate complex in a lobe-specific manner. RT-PCR andWestern blot analyses indicate that rat nongastric H-K-ATPase-subunit (_ng) mRNA and protein are present incoagulating gland (anterior prostate) and lateral and dorsalprostate and absent from ventral lobe, whereas Na-K-ATPase -subunit is present in all lobes. RT-PCR analysis shows that Na-K-ATPase ₄ and ₃ and gastricH-K-ATPase -subunit are not present in significant amounts in allprostate lobes. Relatively low levels of Na-K-ATPase ₂were found in lateral, dorsal, and anterior lobes. _ngprotein expression is anteriodorsolateral: highest in coagulatinggland, somewhat lower in dorsal lobe, and even lower in lateral lobe.Na-K-ATPase protein abundance has the reverse order: expression inventral lobe is higher than in coagulating gland. _ngprotein abundance is higher in coagulating gland than distal colonmembranes. Immunohistochemistry shows that in rat and mouse coagulatinggland epithelium _ng protein has an apical polarizationand Na-K-ATPase ₁ is localized in basolateral membranes.The presence of nongastric H-K-ATPase in rodent prostate apicalmembranes may indicate its involvement in potassium concentrationregulation in secretions of these glands.

     

Potassium and sodium balance in U937 cells during apoptosis with and without cell shrinkage.

Staurosporine (STS) and etoposide (Eto) induced apoptosis of the human histiocytic lymphoma cells U937 were studied to determine the role of monovalent ions in apoptotic cell shrinkage. Cell shrinkage, defined as cell dehydration, was assayed by measurement of buoyant density of cells in continuous Percoll gradient. The K+ and Na+ content in cells of different density fractions was estimated by flame emission analysis. Apoptosis was evaluated by confocal microscopy and flow cytometry of acridine orange stained cells, by flow DNA cytometry and by effector caspase activity. Apoptosis of U937 cells induced by 1 muM STS for 4 h was found to be paralleled by an increase in buoyant density indicating cell shrinkage. An increase in density was accompanied by a decrease in K+ content (from 1.1 to 0.78 mmol/g protein), which exceeded the increase in Na+ content (from 0.30 to 0.34 mmol/g) and resulted in a significant decrease of the total K+ and Na+ content (from 1.4 to 1.1 mmol/g). In contrast to STS, 50 microM Eto for 4 h or 0.8-8 microM Eto for 18-24 h induced apoptosis without triggering cell shrinkage. During apoptosis of U937 cells induced by Eto the intracellular K(+)/Na+ ratio decreased like in the cells treated with STS, but the total K+ and Na+ content remained virtually the same due to a decrease in K+ content being nearly the same as an increase in Na+ content. Apoptotic cell dehydration correlated with the shift of the total cellular K+ and Na+ content. There was no statistically significant decrease in K+ concentration per cell water during apoptosis induced by either Eto (by 13.5%) or STS (by 8%), whereas increase in Na+ concentration per cell water was statistically significant (by 27% and 47%, respectively). The data show that apoptosis can occur without cell shrinkage-dehydration, that apoptosis with shrinkage is mostly due to a decrease in cellular K+ content, and that this decrease is not accompanied by a significant decrease of K+ concentration in cell water.     

Revision of the Athyrisininae, Siluro-devonian Brachiopods from China and Russia

Athyrisinine brachiopods from the Upper Silurian of Russia and the Lower–Middle Devonian of China and north Vietnam include over 70 species belonging to Athyrisina and Parathyrisina ; generic and subfamilial diagnoses are emended. Five genera are considered synonyms of Athyrisina and three of Parathyrisina . A neotype is selected and illustrated for Athyrisina squamosa , the type species of Athyrisina . Four nomina nova , Athyrisina xui , Parathyrisina wani , P. minima , and P. cheni , are suggested as new substitute names for Athyrisina tumida Wang, in Xu et al. 1978 (primary homonymy), Athyrisinoides ganxiensis Wan, in Xu et al. 1978, Parathyrisina minor Zhang, in Zhang and Fu 1983, and Athyrisinoides tudilingensis Chen, 1979 (secondary homonyms) respectively. A new genus, Bruntosina , is described from the Emsian to Eifelian of the Qinling region. A new subfamily, Homeathyridinae, is erected within the Athyrididae with Homeathyris , Pseudohomeospira , and Squamathyris included, revised and their diagnoses emended. Homeathyris incisus sp. nov. is described from the Ludfordian of Novaya Zemlya. Ikella is revised and excluded from the athyrisinins. The origin and dispersal of the homeathyridins and the athyrisinins are discussed.     

Selective Cytostatic and Neurotoxic Effects of Avermectins and Activation of the GABAα Receptors

A natural avermectin complex, aversectin C, was shown to be capable of exerting selective cytostatic and neurotoxic effects on mammalian cells. Specifically, it killed proliferating neuroblastoma B103 cells but was non-toxic for differentiated cells of this culture. The anti-proliferation action of aversectin C was not inhibited by bicuculline or picrotoxin, antagonists of the GABA receptors, and was partly due to the action of avermectin A₁, a component of aversectin C. Aversectin C irreversibly suppressed activity of 60% neurons in medial septal slices of the rat brain. More than 55% of them were the GABA- and B₁-sensitive neurons whereas the rest, about 45% neurons, were the GABA-insensitive and the neurotoxic effect of aversectin C was caused mainly by the B₂ component.     

Historic DNA reveals genetic consequences of fragmentation in an endangered,endemic mustard

Understanding how anthropogenic disturbance affects genetic diversity is essential to appropriately incorporating genetic considerations into conservation plans. Unfortunately, we rarely have information about a population’s genetic diversity before it becomes imperiled. Here we reconstruct the historic range of the naturally rare annual mustard Streptanthus glandulosus subsp. niger (Sgn) and use herbarium specimens to quantify pre-disturbance genetic diversity. We compare this to the genetic diversity in the contemporary plant populations and to plants in the seed bank. We conclude that Sgn was recently a single, panmictic population composed of orders of magnitude more plants than exist today but experienced recent and abrupt declines following housing development. Today Sgn persists as two disjunct populations, the larger of which has retained historic levels of diversity although there is a downward trend in all measures. The smaller population has lost 21–28% of the diversity that was present only 50 years ago with an N_e?~?5–16. The contemporary populations have differentiated from each other due to drift. The seed bank contained no novel alleles and had high levels of homozygosity, indicating that it is incapable of providing genetic rescue. This novel combination of hDNA, the aboveground plant population and the seed bank can be used to design high impact conservation plans that appropriately incorporate genetic diversity for this and other imperiled species.

     

The genius of E. E. metchnikoff— Discoveries over the centuries

On the 15 of May we celebrated the 150th anniversary of the outstanding Russian biologist Elias E. Metchnikoff (1845–1916)—Nobel Prize winner (1908), full and honorary member of many scientific academies of the world. His main works were applied to the zoology of invertebtates, evolution, embryology, immunology, microbiology, infectious pathology, gerontology, etc. Elias Metchnikoff published essays on anthropology, theory of orthobiosis, role of social and social-hygienic factors in solving the problems of old age and life elongation. On 30 May-2 June 1995 an International Symposium dedicated to Metchnikoff's 150th anniversary was held in Moscow. This is a text of the lecture given by us at the opening ceremony.     

Exo70 stimulates the arp2/3 complex for lamellipodia formation and directional cell migration

Directional cell migration requires the coordination of actin assembly and membrane remodeling. The exocyst is an octameric protein complex essential for exocytosis and plasma membrane remodeling [1, 2]. A component of the exocyst, Exo70, directly interacts with the Arp2/3 complex, a core nucleating factor for the generation of branched actin networks for cell morphogenesis and migration [3-9]. Using in?vitro actin polymerization assay and time-lapse total internal reflection fluorescence microscopy, we found that Exo70 functions as a kinetic activator of the Arp2/3 complex that promotes actin filament nucleation and branching. We further found that the effect of Exo70 on actin is mediated by promoting the interaction of the Arp2/3 complex with WAVE2, a member of the N-WASP/WAVE family of nucleation promoting factors. At the cellular level, the stimulatory effect of Exo70 on the Arp2/3 complex is required for lamellipodia formation and maintaining directional persistence of cell migration. Our findings provide a novel mechanism for regulating actin polymerization and branching for effective membrane protrusion during cell morphogenesis and migration.     

Growth pattern, tuber formation and hormonal balance in in vitro potato plants carrying ipt gene

Nodal cuttings of in vitro grown potato plants (Solanum tuberosum, cv. Miranda) were transformed by a vector plasmid carrying ipt gene of Agrobacterium tumefaciens. From the initial teratoma stage 5 clones of transgenic plants (1, 2, 11, 13 and 15) were obtained, which displayed in varying degree shortening of the internodes, decrease of the leaf size, decrease of apical dominance and poor rooting. In addition, two of the clones (11 and 13) showed increased stolon and tuber formation. In all these clones the endogenous level of free cytokinins (CKs) was increased: from 40% in clone 11 to almost 300% in clone 1. Also free indole-3-acetic acid (IAA) level was increased, but to a lower degree; the maximal increase was 160% (clone 13). Applied kinetin or IAA (1 mg·l^-1) strongly suppressed root and tuber formation in clones 11 and 13, although they did not affect or even stimulated these processes in control plants. For control plants the minimal medium sucrose concentration necessary for tuber initiation was 6% whereas in clone 11 plants 2% was sufficient. Different distribution of endogenous CKs and IAA was observed in clone 11 and control plants. The highest CK content was found in transgenic plants in stems and in controls in leaves. In clone 11 plants abscisic acid (ABA) level was significantly increased in comparison to the control throughout the cultivation period. Ethylene formation was strongly increased the first week after the subcultivation and later on the difference between transgenic and control plants rapidly diminished. Reactions of clone 11 plants to red (RL) and blue light (BL) were similar to reactions of control plants. In RL clone 11 plants were tall and thin with stunted leaves; in BL they had a teratoma-like appearance and formed a very high number of tubers. The role of hormones in these changes in growth and tuber formation is discussed.