Cellular and subcellular localization of the vasopressin- regulated urea transporter in rat kidney.

The renal urea transporter (RUT) is responsible for urea accumulation in the renal medulla, and consequently plays a central role in the urinary concentrating mechanism. To study its cellular and subcellular localization, we prepared affinity-purified, peptide-derived polyclonal antibodies against rat RUT based on the cloned cDNA sequence. Immunoblots using membrane fractions from rat renal inner medulla revealed a solitary 97-kDa band. Immunocytochemistry demonstrated RUT labeling of the apical and subapical regions of inner medullary collecting duct (IMCD) cells, with no labeling of outer medullary or cortical collecting ducts. Immunoelectron microscopy directly demonstrated labeling of the apical plasma membrane and of subapical intracellular vesicles of IMCD cells, but no labeling of the basolateral plasma membrane. Immunoblots demonstrated RUT labeling in both plasma membrane and intracellular vesicle-enriched membrane fractions from inner medulla, a subcellular distribution similar to that of the vasopressin-regulated water channel, aquaporin-2. In the outer medulla, RUT labeling was seen in terminal portions of short-loop descending thin limbs. Aside from IMCD and descending thin limbs, no other structures were labeled in the kidney. These results suggest that: (i) the RUT provides the apical pathway for rapid, vasopressin-regulated urea transport in the IMCD, (ii) collecting duct urea transport may be increased by vasopressin by stimulation of trafficking of RUT-containing vesicles to the apical plasma membrane, and (iii) the rat urea transporter may provide a pathway for urea entry into the descending limbs of short-loop nephrons.

Estudo experimental do fenômeno de vibração induzida por vórtices em cilindro rígido livre para oscilar com dois graus de liberdade.

O fenômeno de vibração induzida por vórtices (VIV) é um problema fundamental dentro da Mecânica dos Fluidos e um exemplo importante de interação fluido-estrutura. Esta tese investiga fenômeno de VIV quando um cilindro rígido, submetido a escoamento uniforme, está livre para oscilar na direção transversal e alinhada com a corrente incidente. A tese foi estruturada ao redor de sete perguntas relacionadas ao fenômeno de VIV: 1) O fenômeno e os resultados experimentais são repetitivos? 2) Como ocorre a transição entre ramos de resposta? 3) Qual é o papel da inércia da estrutura oscilante? 4) Qual é o papel de sua rigidez? 5) Quais são as frequências naturais mais importantes da estrutura? 6) Quais padrões de esteira se desenvolvem para VIV com dois graus de liberdade? 7) Quais são os efeitos do movimento na direção alinhada com a corrente no processo de formação e desprendimento de vórtices? O fenômeno de VIV é estudado de maneira experimental em uma base elástica pendular capaz de oscilar com o mesmo momento de inércia e frequência natural nas duas direções. Os experimentos de VIV foram realizados em canal de água recirculante e com diferentes condições de inércia e rigidez. A técnica de velocimetria por imagem de partículas foi usada e permitiu identificar diferentes padrões de esteira de vórtices. Verificou-se que o VIV é repetitivo a nível de amplitudes médias e frequências dominantes. A transição dos ramos pode ocorrer de maneira intermitente ou com histerese. Os parâmetros de inércia e rigidez da estrutura são capazes de mudar o regime de oscilação e, para algumas condições, suprimir as vibrações alinhadas com a corrente. Dentre os padrões de esteira observados, um deles não havia sido relatado na literatura e é definido nesta tese. O novo modo de emissão apresenta dois vórtices com circulação oposta e elevada intensidade emitidos por ciclo. A influência da direção alinhada com o escoamento está relacionada a dois efeitos: a velocidade relativa entre o cilindro e o fluido, responsável pelo aumento da circulação dos vórtices na esteira, e o ângulo de fase do movimento nas direções alinhada e transversal, capaz de mudar o processo de formação dos vórtices.

Boron nutrition and chilling tolerance of warm climate crop species

Background Field observations and glasshouse studies have suggested links between boron (B)-deficiency and leaf damage induced by low temperature in crop plants, but causal relationships between these two stresses at physiological, biochemical and molecular levels have yet to be explored. Limited evidence at the whole-plant level suggests that chilling temperature in the root zone restricts B uptake capacity and/or B distribution/utilization efficiency in the shoot, but the nature of this interaction depends on chilling tolerance of species concerned, the mode of low temperature treatment (abrupt versus gradual temperature decline) and growth conditions (e.g. photon flux density and relative humidity) that may exacerbate chilling stress. Scope This review explores roles of B nutrition in chilling tolerance of continual root or transient shoot chills in crop species adapted to warm season conditions. It reviews current research on combined effects of chilling temperature (ranging from > 0 to 20 degrees C) and B deficiency on growth and B nutrition responses in crop species differing in chilling tolerance. Conclusion For subtropical/tropical species (e.g. cucumber, cassava, sunflower), root chilling at 10-17 degrees C decreases B uptake efficiency and B utilization in the shoot and increases the shoot : root ratio, but chilling-tolerant temperate species (e.g. oilseed rape, wheat) require much lower root chill temperatures (2-5 degrees C) to achieve the same responses. Boron deficiency exacerbates chilling injuries in leaf tissues, particularly under high photon flux density. Suggested mechanisms for B x chilling interactions in plants are: (a) chilling-induced reduction in plasmalemma hydraulic conductivity, membrane fluidity, water channel activity and root pressure, which contribute to the decrease in root hydraulic conductance, water uptake and associated B uptake; (b) chilling-induced stomatal dysfunction affecting B transport from root to shoot and B partitioning in the shoot; and (c) B deficiency induced sensitivity to photo-oxidative damage in leaf cells. However, specific evidence for each of the mechanisms is still lacking. Impacts of B status on chilling tolerance in crop species have important implications for the management of B supply during sensitive stages of growth, such as early growth after planting and early reproductive development, both of which can coincide with the occurrence of chilling temperatures in the field.

Exceptional overproduction of a functional human membrane protein

Eukaryotic-especially human-membrane protein overproduction remains a major challenge in biochemistry. Heterologously overproduced and purified proteins provide a starting point for further biochemical, biophysical and structural studies, and the lack of sufficient quantities of functional membrane proteins is frequently a bottleneck hindering this. Here, we report exceptionally high production levels of a correctly folded and crystallisable recombinant human integral membrane protein in its active form; human aquaporin 1 (hAQP1) has been heterologously produced in the membranes of the methylotrophic yeast Pichia pastoris. After solubilisation and a two step purification procedure, at least 90 mg hAQP1 per liter of culture is obtained. Water channel activity of this purified hAQP1 was verified by reconstitution into proteoliposomes and performing stopped-flow vesicle shrinkage measurements. Mass spectrometry confirmed the identity of hAQP1 in crude membrane preparations, and also from purified protein reconstituted into proteoliposomes. Furthermore, crystallisation screens yielded diffraction quality crystals of untagged recombinant hAQP1. This study illustrates the power of the yeast P. pastoris as a host to produce exceptionally high yields of a functionally active, human integral membrane protein for subsequent functional and structural characterization. © 2007 Elsevier Inc. All rights reserved.

Entrapment of a Water Wire in a Hydrophobic Peptide Channel with an Aromatic Lining

A one-dimensional water wire has been characterized by X-ray diffraction in single crystals of the tripeptide Ac-Phe-Pro-Trp-OMe. Crystals in the hexagonal space group P6(5) reveal a central hydrophobic channel lined by aromatic residues which entraps an approximately linear array of hydrogen bonded water molecules. The absence of any significant van der Waals contact with the channel walls suggests that the dominant interaction between the ``water wire'' and ``peptide nanotube'' is electrostatic in origin. An energy difference of 16 KJmol(-1) is estimated for the distinct orientations of the water wire dipole with respect to the macrodipole of the peptide nanotube. The structural model suggests that Grotthuss type proton conduction may, through constricted hydrophobic channels, be facilitated by concerted, rotational reorientation of water molecules.

ROV-based Deep Water Monitoring of the Northern Channel Islands Marine Protected Areas Annual Report - 2007

Research cruises were conducted in August-October 2007 to complete the third annual remotely operated vehicle (ROV)-based assessments of nearshore rocky bottom finfish at ten sites in the northern Channel Islands. Annual surveys at the Channel Islands have been conducted since 2004 at four sites and were expanded to ten sites in 2005 to monitor potential marine protected area (MPA)effects on baseline fish density. Six of the ten sites are in MPAs and four in nearby fished reference areas. In 2007 the amount of soft-only substrate on the 141 track lines surveyed was again estimated in real-time in order to target rocky bottom habitat. These real-time estimates of hard and mixed substrate for all ten sites averaged 57%, 1% more than the post-processed average of 56%. Surveys generated 69.9 km of usable video for use in finfish density calculations, with target rocky bottom habitat accounting for 56% (39.1 km) for all sites combined. The amount of rocky habitat sampled by site averaged 3.8 km and ranged from 3.3 km sampled at South Point, a State Marine Reserve (SMR) off Santa Rosa Island, to 4.7 km at Anacapa Island SMR. A sampling goal of 75 transects at all 10 sites was met using real-time habitat estimates combined with precautionary over-sampling by 10%. A total of seventy kilometers of sampling is projected to produce at least seventy-five 100 m2 transects per site. Thirteen of 26 finfish taxa observed were selected for quantitative evaluation over the time series based on a minimum criterion of abundance (0.05/100 m2). Ten of these 13 finfish appear to be more abundant at the state marine reserves relative to fished areas when densities were averaged across the 2005 to 2007 period. One of the species that appears to be more abundant in fished areas was señorita, a relatively small prey species that is not a commercial or recreational target. (PDF contains 83 pages.)

Channel Islands Deep Water Monitoring Plan Development Workshop Report April 26-27, 2005 University of California, Santa Barbara

In 2003, twelve marine protected areas were established in state waters (0-3 nmi) surrounding the Channel Islands. NOAA is considering extending this network (3-6 nmi) into deeper waters of the Channel Islands National Marine Sanctuary (CINMS). In order for effective long-term management of the deep water reserves to occur, a well-structured monitoring program is required to assess effectiveness. The CINMS and the National Marine Sanctuary Program (NMSP) hosted a 2-day workshop in April 2005 to develop a monitoring plan for the proposed federal marine reserves in that sanctuary. Conducted at the University of California at Santa Barbara, participants included scientists from academic, state, federal, and private research institutions. Workshop participants developed project ideas that could answer priority questions posed by the NMSP. This workshop report will be used to develop a monitoring plan for the reserves. (PDF contains 47 pages.)

Aspects of the washout of salmonid eggs. 1. Observations on the physical characteristics of real and artificial eggs, their rate of fall in a water column and their pattern of settlement in an experimental channel

When salmonid redds are disrupted by spates, the displaced eggs will drift downstream. The mean distance of travel, the types of locations in which the eggs resettle and the depth of reburial of displaced eggs are not known. Investigation of these topics under field conditions presents considerable practical problems, though the use of artificial eggs might help to overcome some of them. Attempts to assess the similarities and/or differences in performance between real and artificial eggs are essential before artificial eggs can validly be used to simulate real eggs. The present report first compares the two types of egg in terms of their measurable physical characteristics (e.g. dimensions and density). The rate at which eggs fall in still water will relate to the rate at which they are likely to resettle in flowing water in the field. As the rate of fall will be influenced by a number of additional factors (e.g. shape and surface texture) which are not easily measured directly, the rates of fall of the two types of egg have been compared directly under controlled conditions. Finally, comparisons of the pattern of settlement of the two types of egg in flowing water in an experimental channel have been made. Although the work was primarily aimed at testing the value of artificial eggs as a simulation of real eggs, several side issues more directly concerned with the properties of real eggs and the likely distance of drift in natural streams have also been explored. This is the first of three reports made on this topic by the author in 1984.

Occurrence and abundance of Acetes shrimps in the Kutubdia channel of Bangladesh coastal water

This investigation was carried out from June ‘89 to May ‘90 and focuses on the occurrence and abundance of Acetes shrimps in the Kutubdia channel. The shrimps of the genus Acetes occurred throughout the year in the area of investigation. Acetes showed a bimodal peak in occurrence, one in late monsoon season (August - September) and other in pre-monsoon season (February - March). But the maximum number of Acetes shrimps was recorded in March (462 individuals/haul). The Acetes population of this channel was dominated by four species, Acetes erythraeus (38.50%), A. indicus (32.98%), A. chinensis (4.48%) and A. japonicus (3.32%).

Water masses and circulation of the Mozambique Channel

From 1977 to 1980, several research cruises were carried out in the coastal waters of Mozambique to collect oceanographic data. The distribution of hydrographic and bathythermograph stations is given. The water masses and circulation were mapped and wind data gathered.

Plankton identified in stomach contents of Oreochromis niloticus (Pisces, Cichlidae) and the water system of Lakes Edward, George, and Kazinga channel - Uganda

The samples were collected from Lake Edward at Rwenshama, Kisenyi and Katwe, and from Lake George at Mahyoro, Kashaka and Kasenyi and in Kazinga Channel at Katunguru. The organisms identified from the water samples obtained irrespective of station or depth were mainly the phytoplankton (diatoms, blue-green algae and green algae). Of the phytoplankton, blue green-algae were the most abundant both in quantity and number of species especially in L. George. In order of importance were Microcystis spp, Planktolyngbya spp and Anabaenopsis spp were the dominant blue greens. Diatoms and green algae were present but less abundant. The estimated proportions of different types of phytoplankton identified in O. niloticus stomach contents indicate that bluegreen algae were the most abundant followed by the diatoms and green algae.

A comparison of the value of viscosity for several water models using Poiseuille flow in a nano-channel.

The viscosity-temperature relation is determined for the water models SPC/E, TIP4P, TIP4P/Ew, and TIP4P/2005 by considering Poiseuille flow inside a nano-channel using molecular dynamics. The viscosity is determined by fitting the resulting velocity profile (away from the walls) to the continuum solution for a Newtonian fluid and then compared to experimental values. The results show that the TIP4P/2005 model gives the best prediction of the viscosity for the complete range of temperatures for liquid water, and thus it is the preferred water model of these considered here for simulations where the magnitude of viscosity is crucial. On the other hand, with the TIP4P model, the viscosity is severely underpredicted, and overall the model performed worst, whereas the SPC/E and TIP4P/Ew models perform moderately.

"Dual-Parallel-Channel" Shape-Gradient Surfaces: Toward Oriented and Reversible Movement of Water Droplets

In this paper, we prepared "dual-parallel-channel" shape-gradient surfaces, on which water droplets can reversibly and orientedly move between two adjacent pools under the guidance of an external voltage. Furthermore, it is found that the motion speed is governed by several parameters, including bath condition, gradient angle, and the working voltage. In this self-transportation process of water droplets, the external voltage works like a traffic light, which can give "moving", "stopping", "turning" and "straight-going" signals to the Water droplets.

Self-assembly of a water chain with tetrameric and decameric clusters in the channel of a mixed-valence (CuCuII)-Cu-I complex

The reaction of Cu(BF4)(2) with pyridine-2,6-dicarboxylic acid (H(2)pydc) and trans-1,2-bis(4-pyridyl)ethylene (bpe) under hydrothermal conditions afforded a porous mixed-valence (CuCuII)-Cu-I coordination polymer. Coexistence of tetrameric and decameric water clusters within the channels of the complex leads to a novel water chain. The metal-organic framework provides both hydrophilic and hydrophobic environments for stabilizing the clusters and retains its integrity upon dehydration and rehydration.