Altered ubiquitination and stability of aquaporin-1 in hypertonic stress

Aquaporin-1 (AQP1) water channel protein expression is increased by hypertonic stress. The contribution of changes in protein stability to hypertonic induction of AQP1 have not been described. Incubation of BALB/c fibroblasts spontaneously expressing AQP1 with proteasome inhibitors increased AQP1 expression, suggesting basal proteasome-dependent degradation of the protein. Degradation by the proteasome is thought to be triggered by polyubiquitination of a target protein. To determine whether AQP1 is ubiquitinated, immunoprecipitation with anti-AQP1 antibodies was performed, and the resultant samples were probed by protein immunoblot for the presence of ubiquitin. Immunoblots demonstrated ubiquitination of AQP1 under control conditions that increased after treatment with proteasome inhibitors (MG132, lactacystin). Exposure of cells to hypertonic medium for as little as 4 h decreased ubiquitination of AQP1, an effect that persisted through 24 h in hypertonic medium. Using metabolic labeling with [35S]methionine, the half-life of AQP1 protein under isotonic conditions was found to be <4 h. AQP1 protein half-life was markedly increased by exposure of cells to hypertonic medium. These observations provide evidence that aquaporins are a target for ubiquitination and proteasome-dependent degradation. Additionally, these studies demonstrate that reduced protein ubiquitination and increased protein stability lead to increased levels of AQP1 expression during hypertonic stress.

Characterization of a Maize Tonoplast Aquaporin Expressed in Zones of Cell Division and Elongation1

We studied aquaporins in maize (Zea mays), an important crop in which numerous studies on plant water relations have been carried out. A maize cDNA, ZmTIP1, was isolated by reverse transcription-coupled PCR using conserved motifs from plant aquaporins. The derived amino acid sequence of ZmTIP1 shows 76% sequence identity with the tonoplast aquaporin γ-TIP (tonoplast intrinsic protein) from Arabidopsis. Expression of ZmTIP1 in Xenopus laevis oocytes showed that it increased the osmotic water permeability of oocytes 5-fold; this water transport was inhibited by mercuric chloride. A cross-reacting antiserum made against bean α-TIP was used for immunocytochemical localization of ZmTIP1. These results indicate that this and/or other aquaporins is abundantly present in the small vacuoles of meristematic cells. Northern analysis demonstrated that ZmTIP1 is expressed in all plant organs. In situ hybridization showed a high ZmTIP1 expression in meristems and zones of cell enlargement: tips of primary and lateral roots, leaf primordia, and male and female inflorescence meristems. The high ZmTIP1 expression in meristems and expanding cells suggests that ZmTIP1 is needed (a) for vacuole biogenesis and (b) to support the rapid influx of water into vacuoles during cell expansion.

High Expression of the Tonoplast Aquaporin ZmTIP1 in Epidermal and Conducting Tissues of Maize1

Aquaporins are integral membrane proteins of the tonoplast and the plasma membrane that facilitate the passage of water through these membranes. Because of their potentially important role in regulating water flow in plants, studies documenting aquaporin gene expression in specialized tissues involved in water and solute transport are important. We used in situ hybridization to examine the expression pattern of the tonoplast aquaporin ZmTIP1 in different organs of maize (Zea mays L.). This tonoplast water channel is highly expressed in the root epidermis, the root endodermis, the small parenchyma cells surrounding mature xylem vessels in the root and the stem, phloem companion cells and a ring of cells around the phloem strand in the stem and the leaf sheath, and the basal endosperm transfer cells in developing kernels. We postulate that the high level of expression of ZmTIP1 in these tissues facilitates rapid flow of water through the tonoplast to permit osmotic equilibration between the cytosol and the vacuolar content, and to permit rapid transcellular water flow through living cells when required.

The human AQP4 gene: definition of the locus encoding two water channel polypeptides in brain.

The aquaporin family of membrane water transport proteins are expressed in diverse tissues, and in brain the predominant water channel protein is AQP4. Here we report the isolation and characterization of the human AQP4 cDNAs and genomic DNA. Two cDNAs were isolated corresponding to the two initiating methionines (M1 in a 323-aa polypeptide and M23 in a 301-aa polypeptide) previously identified in rat [Jung, J.S., Bhat, R.V., Preston, G.M., Guggino, W.B. & Agre, P. (1994) Proc. Natl. Acad. Sci. USA 91, 13052-13056]. Similar to other aquaporins, the AQP4 gene is composed of four exons encoding 127, 55, 27, and 92 amino acids separated by introns of 0.8, 0.3, and 5.2 kb. Unlike other aquaporins, an alternative coding initiation sequence (designated exon 0) was located 2.7 kb upstream of exon 1. When spliced together, M1 and the subsequent 10 amino acids are encoded by exon 0; the next 11 amino acids and M23 are encoded by exon 1. Transcription initiation sites have been mapped in the proximal promoters of exons 0 and 1. RNase protection revealed distinct transcripts corresponding to M1 and M23 mRNAs, and AQP4 immunoblots of cerebellum demonstrated reactive polypeptides of 31 and 34 kDa. Using a P1 and a lambda EMBL subclone, the chromosomal site of the human AQP4 gene was mapped to chromosome 18 at the junction of q11.2 and q12.1 by fluorescence in situ hybridization. These studies may now permit molecular characterization of AQP4 during human development and in clinical disorders.

A biologic function for an "orphan" messenger: D-myo-inositol 3,4,5,6-tetrakisphosphate selectively blocks epithelial calcium-activated chloride channels.

Inositol phosphates are a family of water-soluble intracellular signaling molecules derived from membrane inositol phospholipids. They undergo a variety of complex interconversion pathways, and their levels are dynamically regulated within the cytosol in response to a variety of agonists. Relatively little is known about the biological function of most members of this family, with the exception of inositol 1,4,5-trisphosphate. Specifically, the biological functions of inositol tetrakisphosphates are largely obscure. In this paper, we report that D-myo-inositol 3,4,5,6-tetrakisphosphate (D-Ins(3,4,5,6)P4) has a direct biphasic (activation/inhibition) effect on an epithelial Ca(2+)-activated chloride channel. The effect of D-Ins(3,4,5,6)P4 is not mimicked by other inositol tetrakisphosphate isomers, is dependent on the prevailing calcium concentration, and is influenced when channels are phosphorylated by calmodulin kinase II. The predominant effect of D-Ins(3,4,5,6)P4 on phosphorylated channels is inhibitory at levels of intracellular calcium observed in stimulated cells. Our findings indicate the biological function of a molecule hitherto considered as an "orphan" messenger. They suggest that the molecular target for D-Ins(3,4,5,6)P4 is a plasma membrane Ca(2+)-activated chloride channel. Regulation of this channel by D-Ins(3,4,5,6)P4 and Ca2+ may have therapeutic implications for the disease states of both diabetic nephropathy and cystic fibrosis.

Influence of characteristics of irrigation channels on the breeding bird community of a wetland zone in southeast of Spain

The “El Hondo Nature Park” is mainly composed of a series of irrigation channels and water reservoirs, subjected to various regimes of management as well as reed and vegetation control, thus creating a great variety of habitats and situations. To determine the influence of these habitats and management regimes on the local bird community, a set of characteristics of these channels and their surrounding area were analysed with a Correspondence Analysis (CA). The degree of reed development in channels and the presence in the surroundings of orchards and other reed formations were the most decisive factors to explain the probability of occurrence of reed birds and waterbirds, as well as bird species richness and abundance. Other bird species were not directly influenced by channel variables, but only by those of surrounding land uses.

Phytoplankton abundance measured on water samples from rivers of the Lena Delta in 2010

The Lena Delta in Northern Siberia is one of the largest river deltas in the world. During peak discharge, after the ice melt in spring, it delivers between 60-8000 m**3/s of water and sediment into the Arctic Ocean. The Lena Delta and the Laptev Sea coast also constitute a continuous permafrost region. Ongoing climate change, which is particularly pronounced in the Arctic, is leading to increased rates of permafrost thaw. This has already profoundly altered the discharge rates of the Lena River. But the chemistry of the river waters which are discharged into the coastal Laptev Sea have also been hypothesized to undergo considerable compositional changes, e.g. by increasing concentrations of inorganic nutrients such as dissolved organic carbon (DOC) and methane. These physical and chemical changes will also affect the composition of the phytoplankton communities. However, before potential consequences of climate change for coastal arctic phytoplankton communities can be judged, the inherent status of the diversity and food web interactions within the delta have to be established. In 2010, as part of the AWI Lena Delta programme, the phyto- and microzooplankton community in three river channels of the delta (Trofimov, Bykov and Olenek) as well as four coastal transects were investigated to capture the typical river phytoplankton communities and the transitional zone of brackish/marine conditions. Most CTD profiles from 23 coastal stations showed very strong stratification. The only exception to this was a small, shallow and mixed area running from the outflow of Bykov channel in a northerly direction parallel to the shore. Of the five stations in this area, three had a salinity of close to zero. Two further stations had salinities of around 2 and 5 throughout the water column. In the remaining transects, on the other hand, salinities varied between 5 and 30 with depth. Phytoplankton counts from the outflow from the Lena were dominated by diatoms (Aulacoseira species) cyanobacteria (Aphanizomenon, Pseudanabaena) and chlorophytes. In contrast, in the stratified stations the plankton was mostly dominated by dinoflagellates, ciliates and nanoflagellates, with only an insignificant diatom component from the genera Chaetoceros and Thalassiosira (brackish as opposed to freshwater species). Ciliate abundance was significantly coupled with the abundance of total flagellates. A pronounced partitioning in the phytoplankton community was also discernible with depth, with a different community composition and abundance above and below the thermocline in the stratified sites. This work is a first analysis of the phytoplankton community structure in the region where Lena River discharge enters the Laptev Sea.

A new method of determinating co-efficients of flow and of resistance in pipes and in open channels, and new formulae for head, diameter, mean hydraulic radius, velocity, etc., based on this new theory /

Mode of access: Internet.

Responsiveness summary : United States Army Corps of Engineers : Waukegan Harbor, Lake County, Illinois : Section 401 water quality certification for dredging of the Waukegan Harbor approach channel and advanced maintenance area.

The approved project allows the Corps to dredge the Waukegan Harbor approach channel and advanced maintenance area. The area to be dredged lies immediately east of the north breakwater. It is approximately a rectangle 650 feet north and south and 1,400 feet east and west. The advanced maintenance area is a band along the north side of the channel. The approved project is for a 10-year certification, under which the Corps may remove 22,000 to 75,000 cubic yards of sediment per dredging event. The dredging depth is 22 feet and the amount to be dredged is about one foot of sediment. As a condition of the certification, disposal of the dredged sediment in Lake Michigan or the waters of the state cannot occur until the conditions of the certification are met. These conditions, which have been placed on the certification by Illinois EPA, ensure that the project meets state water quality standards and is consistent with the determinations of the Illinois Attorney General's Task Force on asbestos contamination at Illinois Beach State Park.

A technique for determining depths for T-year discharges in rigid-boundary channels /

Bibliography: p. 38.

Structures of mu O-conotoxins from Conus marmoreus - Inhibitors of tetrodotoxin (TTX)-sensitive and TTX-resistant sodium channels in mammalian sensory neurons

The muO-conotoxins are an intriguing class of conotoxins targeting various voltage-dependent sodium channels and molluscan calcium channels. In the current study, we have shown MrVIA and MrVIB to be the first known peptidic inhibitors of the transient tetrodotoxin-resistant (TTX-R) Na+ current in rat dorsal root ganglion neurons, in addition to inhibiting tetrodotoxin-sensitive Na+ currents. Human TTX-R sodium channels are a therapeutic target for indications such as pain, highlighting the importance of the muO-conotoxins as potential leads for drug development. Furthermore, we have used NMR spectroscopy to provide the first structural information on this class of conotoxins. MrVIA and MrVIB are hydrophobic peptides that aggregate in aqueous solution but were solubilized in 50% acetonitrile/water. The three-dimensional structure of MrVIB consists of a small beta-sheet and a cystine knot arrangement of the three-disulfide bonds. It contains four backbone loops between successive cysteine residues that are exposed to the solvent to varying degrees. The largest of these, loop 2, is the most disordered part of the molecule, most likely due to flexibility in solution. This disorder is the most striking difference between the structures of MrVIB and the known delta- and omega-conotoxins, which along with the muO-conotoxins are members of the O superfamily. Loop 2 of omega-conotoxins has previously been shown to contain residues critical for binding to voltage-gated calcium channels, and it is interesting to speculate that the flexibility observed in MrVIB may accommodate binding to both sodium and molluscan calcium channels.

Year-round presence of dugongs in Pumicestone Passage, south-east Queensland, examined in relation to water temperature and seagrass distribution

Pumicestone Passage is a narrow waterway that lies to the north of and adjacent to Moreton Bay, and between mainland Queensland and Bribie Island, Australia. Anecdotal reports have suggested that the Passage is home to dugongs year-round despite winter water temperatures that are known to cause dugongs to migrate elsewhere. To examine the pattern of distribution and abundance of dugongs within the passage on a year-round basis, eight years of sightings data collected by a charter boat operator were examined. Dedicated aerial surveys of the passage were also conducted at two-monthly intervals over two years, and more intensively over a single winter. Dugong sightings were examined in relation to water temperatures and seagrass prevalence. The number of dugongs sighted in the area on any one survey varied from 0 to 13. Dugongs were seen in all months of the year and in each of the eight winters, indicating that Pumicestone Passage is used year-round despite winter water temperatures dropping to below 18 degrees C from June to August inclusive and below 16 degrees C in June. All dugong sightings occurred in the southern part of the passage, south of Tripcony Bight. Dugongs were associated with shallows that support Halophila and Halodule species of seagrass, food species that are favoured elsewhere in their range. The northern part of the passage also supports seagrasses that are eaten by dugongs and has water temperature ranges that are not appreciably different to those of the southern passage. However, the narrow channels and very shallow nature of the northern passage provides little to no deep-water refugia for dugongs and the seagrass beds are less extensive. This study suggests that southern Pumicestone Passage requires protection concomitant with it being a year-round refuge of the vulnerable dugong.

Modeling of Flowing Film Concentrators - Part 1. Water Split Behaviour

A large number of mineral processing equipment employs the basic principles of gravity concentration in a flowing fluid of a few millimetres thick in small open channels where the particles are distributed along the flow height based on their physical properties and the fluid flow characteristics. Fluid flow behaviour and slurry transportation characteristics in open channels have been the research topic for many years in many engineering disciplines. However, the open channels used in the mineral processing industries are different in terms of the size of the channel and the flow velocity used. Understanding of water split behaviour is, therefore, essential in modeling flowing film concentrators. In this paper, an attempt has been made to model the water split behaviour in an inclined open rectangular channel, resembling the actual size and the flow velocity used by the mineral processing industries, based on the Prandtl's mixing length approach. (c) 2006 Elsevier B.V. All rights reserved.

Membrane trafficking of aquaporin 1 is mediated by protein kinase C via microtubules and regulated by tonicity

It is well-known that the rapid flow of water into and out of cells is controlled by membrane proteins called aquaporins (AQPs). However, the mechanisms that allow cells to quickly respond to a changing osmotic environment are less well established. Using GFP-AQP fusion proteins expressed in HEK293 cells, we demonstrate the reversible manipulation of cellular trafficking of AQP1. AQP1 trafficking was mediated by the tonicity of the cell environment in a specific PKC- and microtubule-dependent manner. This suggests that the increased level of water transport following osmotic change may be due a phosphorylation-dependent increase in the level of AQP1 trafficking resulting in membrane localization.

The separation of secondary oil-water dispersions in particulate beds

The literature relating to haze formation, methods of separation, coalescence mechanisms, and models by which droplets <100 μm are collected, coalesced and transferred, have been reviewed with particular reference to particulate bed coalescers. The separation of secondary oil-water dispersions was studied experimentally using packed beds of monosized glass ballotini particles. The variables investigated were superficial velocity, bed depth, particle size, and the phase ratio and drop size distribution of inlet secondary dispersion. A modified pump loop was used to generate secondary dispersions of toluene or Clairsol 350 in water with phase ratios between 0.5-6.0 v/v%.Inlet drop size distributions were determined using a Malvern Particle Size Analyser;effluent, coalesced droplets were sized by photography. Single phase flow pressure drop data were correlated by means of a Carman-Kozeny type equation. Correlations were obtained relating single and two phase pressure drops, as (ΔP2/μc)/ΔP1/μd) = kp Ua Lb dcc dpd Cine A flow equation was derived to correlate the two phase pressure drop data as, ΔP2/(ρcU2) = 8.64*107 [dc/D]-0.27 [L/D]0.71 [dp/D]-0.17 [NRe]1.5 [e1]-0.14 [Cin]0.26  In a comparison between functions to characterise the inlet drop size distributions a modification of the Weibull function provided the best fit of experimental data. The general mean drop diameter was correlated by: q_p q_p p_q /β      Γ ((q-3/β) +1) d qp = d fr  .α        Γ ((P-3/β +1 The measured and predicted mean inlet drop diameters agreed within ±15%. Secondary dispersion separation depends largely upon drop capture within a bed. A theoretical analysis of drop capture mechanisms in this work indicated that indirect interception and London-van der Waal's mechanisms predominate. Mathematical models of dispersed phase concentration m the bed were developed by considering drop motion to be analogous to molecular diffusion.The number of possible channels in a bed was predicted from a model in which the pores comprised randomly-interconnected passage-ways between adjacent packing elements and axial flow occured in cylinders on an equilateral triangular pitch. An expression was derived for length of service channels in a queuing system leading to the prediction of filter coefficients. The insight provided into the mechanisms of drop collection and travel, and the correlations of operating parameters, should assist design of industrial particulate bed coalescers.