Miniaturization of the Clonogenic Assay Using Confluence Measurement

PubMed Central

Mayr, Christian; Beyreis, Marlena; Dobias, Heidemarie; Gaisberger, Martin; Pichler, Martin; Ritter, Markus; Jakab, Martin; Neureiter, Daniel; Kiesslich, Tobias

2018-01-01

The clonogenic assay is a widely used method to study the ability of cells to ‘infinitely’ produce progeny and is, therefore, used as a tool in tumor biology to measure tumor-initiating capacity and stem cell status. However, the standard protocol of using 6-well plates has several disadvantages. By miniaturizing the assay to a 96-well microplate format, as well as by utilizing the confluence detection function of a multimode reader, we here describe a new and modified protocol that allows comprehensive experimental setups and a non-endpoint, label-free semi-automatic analysis. Comparison of bright field images with confluence images demonstrated robust and reproducible detection of clones by the confluence detection function. Moreover, time-resolved non-endpoint confluence measurement of the same well showed that semi-automatic analysis was suitable for determining the mean size and colony number. By treating cells with an inhibitor of clonogenic growth (PTC-209), we show that our modified protocol is suitable for comprehensive (broad concentration range, addition of technical replicates) concentration- and time-resolved analysis of the effect of substances or treatments on clonogenic growth. In summary, this protocol represents a time- and cost-effective alternative to the commonly used 6-well protocol (with endpoint staining) and also provides additional information about the kinetics of clonogenic growth. PMID:29510509

Regulation of human corneal epithelial mucins by rebamipide.

PubMed

Itoh, Shinsaku; Itoh, Kuni; Shinohara, Hisashi

2014-02-01

Membrane-associated mucins (MAMs) play important roles in barrier function and tear stability, and their expression on the ocular surface is altered in dry eye disease. Rebamipide is a mucin secretagogue that promotes the production of mucin-like glycoproteins in human corneal epithelial (HCE) cells. However, the expression of MAMs on the corneal epithelia (MUC1, MUC4, MUC16), which is induced by rebamipide, is poorly understood. In this study, we investigated the effect of rebamipide on the regulation of MAM expression in HCE cells. MUC16, Ki67 and PCNA expression levels in HCE cells isolated at confluence and at 24 hours after confluence were examined by Western blotting to assess cell proliferation. HCE cells isolated at 24 hours after confluence were cultured in medium supplemented with 1-10 µM rebamipide or 0.3-30 nM of epidermal growth factor (EGF). Real-time PCR (RT-PCR) and Western blot analysis of MAMs were performed to evaluate the effect of rebamipide. Western blot analysis of cells treated with an EGF receptor inhibitor (AG1478) or MEK1/2 inhibitor (U0126) was performed to reveal the relationship between EGF receptor activation and rebamipide-induced MAM expression. HCE cells isolated at 24 hours after confluence had lower cell proliferation activity and increased MUC16 expression compared with cells isolated at confluence. RT-PCR and Western blot analysis revealed that rebamipide increased MAM gene expression for 2 hours and protein expression for 24 hours in HCE cells. EGF inhibitor treatment led to reduced levels of all three MAMs that are normally induced by rebamipide, whereas EGF induced the expression of all three MAMs. We suggested that rebamipide increased MUC1, MUC4 and MUC16 expression levels through signals involved in EGF receptor activation in the human corneal epithelia. These data suggest that rebamipide may improve subjective symptoms of dry eye disease by upregulating MAM expression.

Maintenance of human adipose derived stem cell (hASC) differentiation capabilities using a 3D culture.

PubMed

Lin, Ching-Yu; Huang, Chi-Hui; Wu, Yuan-Kun; Cheng, Nai-Chen; Yu, Jiashing

2014-07-01

In this study, 3D culture system for human adipose-derived stem cell (hASC) using a BioLevitator as the bioreactor for microcarrier-based cultures was established. During the culturing period, hASCs preferred to grow in crevices between microcarriers and a high viability was maintained even when reaching confluency. Adipogenic or osteogenic differential medium was used to induce hASCs and differential potentials of these cells were compared between 2D and 3D environments via RT-PCR and staining quantifications. CEBP/α gene expression was significant higher in 3D condition at day 21 (P < 0.05). Staining quantification indicates that cells cultured in 3D condition have significant better differentiation potential from day 14 to 21 for both adipogenic and osteogenic lineages (P < 0.01).

Renal cysteine conjugate C-S lyase mediated toxicity of halogenated alkenes in primary cultures of human and rat proximal tubular cells.

PubMed

McGoldrick, Trevor A; Lock, Edward A; Rodilla, Vicente; Hawksworth, Gabrielle M

2003-07-01

Proximal tubular cells from human (HPT) and rat (RPT) kidneys were isolated, grown to confluence and incubated with S-(1,2-dichlorovinyl)- l-cysteine (DCVC), S-(1,2,2-trichlorovinyl)- l-cysteine (TCVC), S-(1,1,2,2-tetrafluoroethyl)- l-cysteine (TFEC) and S-(2-chloro-1,1-difluorethyl)- l-cysteine (CDFEC), the cysteine conjugates of nephrotoxicants. The cultures were exposed to the conjugates for 12, 24 and 48 h and the toxicity determined using the MTT assay. All four conjugates caused dose-dependent toxicity to RPT cells over the range 50-1,000 microM, the order of toxicity being DCVC>TCVC>TFEC=CDFEC. The inclusion of aminooxyacetic acid (AOAA; 250 microM), an inhibitor of pyridoxal phosphate-dependent enzymes such as C-S lyase, afforded protection, indicating that C-S lyase has a role in the bioactivation of these conjugates. In HPT cultures only DCVC caused significant time- and dose-dependent toxicity. Exposure to DCVC (500 microM) for 48 h decreased cell viability to 7% of control cell values, whereas co-incubation of DCVC (500 microM) with AOAA (250 microM) resulted in cell viability of 71%. Human cultures were also exposed to S-(1,2-dichlorovinyl)-glutathione (DCVG). DCVG was toxic to HPT cells, but the onset of toxicity was delayed compared with the corresponding cysteine conjugate. AOAA afforded almost complete protection from DCVG toxicity. Acivicin (250 microM), an inhibitor of gamma-glutamyl transferase (gamma-GT), partially protected against DCVG (500 microM)-induced toxicity at 48 h (5% viability and 53% viability in the absence and presence of acivicin, respectively). These results suggest that DCVG requires processing by gamma-GT prior to bioactivation by C-S lyase in HPT cells. The activity of C-S lyase, using TFEC as a substrate, and glutamine transaminase K (GTK) was measured in rat and human cells with time in culture. C-S lyase activity in RPT and HPT cells decreased to approximately 30% of fresh cell values by the time the cells reached confluence (120 h), whereas the decline in GTK activity was less marked (50% of the fresh cell values at confluence). Rat cells had threefold higher activity than human cells at each time point. This higher activity may partly explain the differences in toxicity between rat and human proximal tubular cells in culture.

Snake River fall Chinook salmon life history investigations, annual report 2008

USGS Publications Warehouse

Tiffan, Kenneth F.; Connor, William P.; Bellgraph, Brian J.; Buchanan, Rebecca A.

2010-01-01

In 2009, we used radio and acoustic telemetry to evaluate the migratory behavior, survival, mortality, and delay of subyearling fall Chinook salmon in the Clearwater River and Lower Granite Reservoir. We released a total of 1,000 tagged hatchery subyearlings at Cherry Lane on the Clearwater River in mid August and we monitored them as they passed downstream through various river and reservoir reaches. Survival through the free-flowing river was high (>0.85) for both radio- and acoustic-tagged fish, but dropped substantially as fish delayed in the Transition Zone and Confluence areas. Estimates of the joint probability of migration and survival through the Transition Zone and Confluence reaches combined were similar for both radio- and acoustic-tagged fish, and ranged from about 0.30 to 0.35. Estimates of the joint probability of delaying and surviving in the combined Transition Zone and Confluence peaked at the beginning of the study, ranging from 0.323 ( SE =NA; radio-telemetry data) to 0.466 ( SE =0.024; acoustic-telemetry data), and then steadily declined throughout the remainder of the study. By the end of October, no live tagged juvenile salmon were detected in either the Transition Zone or the Confluence. As estimates of the probability of delay decreased throughout the study, estimates of the probability of mortality increased, as evidenced by the survival estimate of 0.650 ( SE =0.025) at the end of October (acoustic-telemetry data). Few fish were detected at Lower Granite Dam during our study and even fewer fish passed the dam before PIT-tag monitoring ended at the end of October. Five acoustic-tagged fish passed Lower Granite Dam in October and 12 passed the dam in November based on detections in the dam tailrace; however, too few detections were available to calculate the joint probabilities of migrating and surviving or delaying and surviving. Estimates of the joint probability of migrating and surviving through the reservoir was less than 0.2 based on acoustic-tagged fish. Migration rates of tagged fish were highest in the free-flowing river (median range = 36 to 43 km/d) but were generally less than 6 km/d in the reservoir reaches. In particular, median migration rates of radio-tagged fish through the Transition Zone and Confluence were 3.4 and 5.2 km/d, respectively. Median migration rate for acoustic-tagged fish though the Transition Zone and Confluence combined was 1 km/d.

[A method for the primary culture of fibroblasts isolated from human airway granulation tissues].

PubMed

Chen, Nan; Zhang, Jie; Xu, Min; Wang, Yu-ling; Pei, Ying-hua

2013-04-01

To establish a feasible method to culture primary fibroblasts isolated from human airway granulation tissues, and therefore to provide experimental data for the investigation of the pathogenesis of benign airway stenosis. The granulation tissues were collected from 6 patients during routine bronchoscopy at our department of Beijing Tiantan Hospital from April to June 2011. Primary fibroblasts were obtained by culturing the explanted tissues. Cell growth was observed under inverted microscope. All of these 6 primary cultures were successful. Fibroblast-like cells were observed to migrate from the tissue pieces 3 d after inoculation. After 9-11 d of culture, cells reached to 90% confluence and could be sub-cultured. After passage, the cells were still in a typical elongated spindle-shape and grew well. The cells could be sub-cultured further when they formed a monolayer. Explant culture is a reliable method for culturing primary fibroblasts from human airway granulation tissues.

[Isolation and culture of bovine choriocapillary endothelial cells using paramagnetic beads coated with Lycopersicon esculentum].

PubMed

Swiech-Zubilewicz, A; Soubrane, G; Mascarelli, F

2000-01-01

To establish a pure culture of choriocapillary endothelial cells as a model of angiogenesis in vitro. Bovine choriocapillary endothelial cells (BCEC) were obtained by the method described by Hoffmann et al. (6) using the polystyrene paramagnetic beads coated with Lycopersicon esculentum, which attach specifically to the rest of fucose on the surface of microvascular endothelial cells. The endothelial characteristic of the cultured cells was evaluated by immunocytochemistry using anti von Willebrand factor and anti-CD 31 antibodies. Proliferation and survival of BCEC were tested using haemacytometer of Mallasez. The purity of obtained BCEC culture was confirmed by positive immunocytochemical staining with anti von Willebrand and anti factor CD 31 antibodies in more than 95% of cells. The proliferation of cells in Endothelial Cell Medium resulted in twofold increase of number of cells during 4-day observation period. After reaching the confluence, the cells continued to proliferate with increase of the cell number by 60% during 4-day observation. The use of paramagnetic beads coated with specific lectine provide a pure isolation of BCEC, which can be maintained in culture with preservation of their characteristic.

Factors controlling the abundance of rainbow trout in the Colorado River in Grand Canyon in a reach utilized by endangered humpback chub

USGS Publications Warehouse

Korman, Josh; Yard, Michael D.; Yackulic, Charles B.

2015-01-01

We estimated the abundance, survival, movement, and recruitment of non-native rainbow trout in the Colorado River in Grand Canyon to determine what controls their abundance near the Little Colorado River (LCR) confluence where endangered humpback chub rear. Over a 3-year period, we tagged more than 70,000 trout and recovered over 8,200 tagged fish. Trout density was highest (10,000-25,000 fish/km) in the reach closest to Glen Canyon Dam where the majority of trout recruitment occurs, and was 30-50-fold lower (200-800 fish/km) in reaches near the LCR confluence ~100 km downstream. The extent of rainbow trout movement was limited with less than 1% of recaptures making movements greater than 20 km. However, due to high trout densities in upstream source areas, this small dispersal rate was sufficient to explain the 3-fold increase in the relatively small population near the LCR. Reducing dispersal rates of trout from upstream sources is the most feasible solution to maintain low densities near the LCR to minimize negative effects of competition and predation on humpback chub.

Patterns of variation in diversity of the Mississippi river microbiome over 1,300 kilometers

PubMed Central

Payne, Jason T.; Millar, Justin J.; Jackson, Colin R.

2017-01-01

We examined the downriver patterns of variation in taxonomic diversity of the Mississippi River bacterioplankton microbiome along 1,300 river kilometers, or approximately one third the total length of the river. The study section included portions of the Upper, Middle, and Lower Mississippi River, confluences with five tributaries draining distinct sub-basins, river cities, and extended stretches without major inputs to the Mississippi. The composition and proportional abundance of dominant bacterial phyla was distinct for free-living and particle-associated cells, and constant along the entire reach, except for a substantial but transient disturbance near the city of Memphis, Tennessee. At a finer scale of taxonomic resolution (operational taxonomic units, OTUs), however, there were notable patterns in downriver variation in bacterial community alpha diversity (richness within a site) and beta diversity (variation in composition among sites). There was a strong and steady increase downriver in alpha diversity of OTUs on suspended particles, suggesting an increase in particle niche heterogeneity, and/or particle colonization. Relatively large shifts in beta diversity of free-living and particle-associated communities occurred following major tributary confluences and transiently at Memphis, while in long stretches between these points diversity typically varied more gradually. We conclude that the Mississippi River possesses a bacterioplankton microbiome distinct in diversity from other large river microbiomes in the Mississippi River Basin, that at major river confluences or urban point sources its OTU diversity may shift abruptly and substantially, presumably by immigration of distinct external microbiomes, but that where environmental conditions are more stable along the downriver gradient, microbiome diversity tends to vary gradually, presumably by a process of successional change in community composition. PMID:28350888

The effects of laser immunotherapy on cancer cell migration

NASA Astrophysics Data System (ADS)

Bahavar, Cody F.; Zhou, Feifan; Hasanjee, Aamr M.; Layton, Elivia; Lam, Anh; Chen, Wei R.; Vaughan, Melville B.

2016-03-01

Laser immunotherapy (LIT) uses laser irradiation and immunological stimulation to target all types of metastases and creates a long-term tumor resistance. Glycated chitosan (GC) is the immunological stimulant used in LIT. Interestingly, GC can act as a surfactant for single-walled carbon nanotubes (SWNTs) to immunologically modify SWNTs. SWNT-GC retains the optical properties of SWNTs and the immunological functions of GC to help increase the selectivity of the laser and create a more optimal immune response. One essential aspect of understanding this immune response is knowing how laser irradiation affects cancer cells' ability to metastasize. In this experiment, a cell migration assay was performed. A 2mm circular elastomer plugs were placed at the bottom of multi-well dishes. Pre-cancerous keratinocytes, different tumor cells, and fibroblasts were then plated separately in treated wells. Once the cells reached 100% confluence, they were irradiated by either a 980nm or 805nm wavelength laser. The goal was to determine the effects of laser irradiation and immunological stimulation on cancer cell migration in vitro, paying the way to understand the mechanism of LIT in treating metastatic tumors in cancer patients.

Production of cloned calves using roscovitine-treated adult somatic cells as donors.

PubMed

Miyoshi, Kazuchika; Arat, Sezen; Stice, Steven L

2006-01-01

The stage of the donor cell cycle is a major factor in the success of cloning. Quiescent cells arrested in the G0/G1 phases of the cell cycle by either serum starvation or growth arrest when cultured cells reach confluence have been used as donors to produce cloned animals. Recently, we have developed a novel and effective method using roscovitine to synchronize adult bovine granulosa cells in the G0/G1 cell cycle stage. The resulting fetal and calf survival after transfer of cloned embryos was enhanced in the roscovitine-treated group compared with serum-starved controls. The methods described in this chapter outline (1) the preparation of donor cells, (2) the preparation of recipient oocytes, and (3) the production of cloned embryos. The first section involves methods for the preparation of donor cell stocks from isolated granulosa cells and the roscovitine treatment of the cells before nuclear transfer. The second section explains procedures of in vitro maturation of recipient oocytes. The last section involves methods for the production of cell-oocyte complexes, the fusion of the complexes, and the activation, in vitro culture, and transfer into recipient females of cloned embryos.

Effects of fish community on occurrences of Yangtze finless porpoise in confluence of the Yangtze and Wanhe Rivers.

PubMed

Zhang, Xiaoke; Yu, Daoping; Wang, Huili; Wan, An; Chen, Minmin; Tao, Feng; Song, Zunrong

2015-06-01

The Yangtze finless porpoise is a subspecies of narrow-ridged finless porpoise endemic to the middle and lower reaches of the Yangtze River and the adjoining Poyang and Dongting Lakes. With the depletion of fish stocks in the Yangtze River in recent decades, food availability has become the most important factor affecting the survival of this subspecies. Despite this, the relationships between fish community and occurrences of porpoise are far from being fully understood. Therefore, during September 2013 to August 2014, the occurrences of porpoise were investigated in confluence of the Yangtze and Wanhe Rivers; fish community was also surveyed synchronously in confluence and two adjacent transects. The results showed that (1) the confluence had maximum fish species richness, and the main dominant species was upper fish, while the other two transects were mainly dominated by demersal fish. ANOVA analyses showed that individual number and yield of upper fish which can be eaten by porpoise (upper edible fish) in the confluence were significantly higher than other two transects. (2) Average group size of the porpoise was 3.7 ± 1.8 individuals. The occurrences of porpoise in different seasons had great differences, and the porpoise was more likely to be detected in autumn and winter. (3) Fish community had significant effects on occurrences of porpoise, and the main influencing factors were fish species richness, individual number, and yield of edible fish, especially the upper edible fish. The results of this study will have important implications for the conservation of porpoise.

75 FR 29258 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-25

... Barker, Town Reach 1. of Chenango, Town of Dickinson, Town of Fenton, Village of Port Dickinson... River. Town of Fenton. Approximately 2.5 miles None +1011 upstream of Ballyhack Road. Page Brook At the confluence with +876 +878 Town of Fenton. the Chenango River. Approximately 630 feet +877 +878 downstream of...

Fabrication of corneal epithelial cell sheets maintaining colony-forming cells without feeder cells by oxygen-controlled method.

PubMed

Nakajima, Ryota; Takeda, Shizu

2014-01-01

The use of murine 3T3 feeder cells needs to be avoided when fabricating corneal epithelial cell sheets for use in treating ocular surface diseases. However, the expression level of the epithelial stem/progenitor cell marker, p63, is down-regulated in feeder-free culture systems. In this study, in order to fabricate corneal epithelial cell sheets that maintain colony-forming cells without using any feeder cells, we investigated the use of an oxygen-controlled method that was developed previously to fabricate cell sheets efficiently. Rabbit limbal epithelial cells were cultured under hypoxia (1-10% O2) and under normoxia during stratification after reaching confluence. Multilayered corneal epithelial cell sheets were fabricated using an oxygen-controlled method, and immunofluorescence analysis showed that cytokeratin 3 and p63 was expressed in appropriate localization in the cell sheets. The colony-forming efficiency of the cell sheets fabricated by the oxygen-controlled method without feeder cells was significantly higher than that of cell sheets fabricated under 20% O2 without feeder cells. These results indicate that the oxygen-controlled method has the potential to achieve a feeder-free culture system for fabricating corneal epithelial cell sheets for corneal regeneration. Copyright © 2013 Elsevier Ltd. All rights reserved.

Quantitative assessment of neural outgrowth using spatial light interference microscopy

NASA Astrophysics Data System (ADS)

Lee, Young Jae; Cintora, Pati; Arikkath, Jyothi; Akinsola, Olaoluwa; Kandel, Mikhail; Popescu, Gabriel; Best-Popescu, Catherine

2017-06-01

Optimal growth as well as branching of axons and dendrites is critical for the nervous system function. Neuritic length, arborization, and growth rate determine the innervation properties of neurons and define each cell's computational capability. Thus, to investigate the nervous system function, we need to develop methods and instrumentation techniques capable of quantifying various aspects of neural network formation: neuron process extension, retraction, stability, and branching. During the last three decades, fluorescence microscopy has yielded enormous advances in our understanding of neurobiology. While fluorescent markers provide valuable specificity to imaging, photobleaching, and photoxicity often limit the duration of the investigation. Here, we used spatial light interference microscopy (SLIM) to measure quantitatively neurite outgrowth as a function of cell confluence. Because it is label-free and nondestructive, SLIM allows for long-term investigation over many hours. We found that neurons exhibit a higher growth rate of neurite length in low-confluence versus medium- and high-confluence conditions. We believe this methodology will aid investigators in performing unbiased, nondestructive analysis of morphometric neuronal parameters.

Flood Plain Information Brandywine Creek, New Castle County, Delaware.

DTIC Science & Technology

1973-07-01

The Brandywine’s course through Delaware is fairly straight and cuts through a valley of rugged terrain. The adjacent land slopes that comprise its...varies sharply in 3 distinct reaches. The upper reach from the State Line to Rockland Road falls 15.5 feet at a slope of 4.9 feet per mile. The middle...Mileage Above Drainage Area Location Mouth Tributary Total sq. Mi. sq. mi. Pennsy lvania-Del aware State Line 10.55 ... 299.9 Confluence with Beaver

Metropolitan Spokane Region Water Resources Study. Appendix J. Water Quality Simulation Model

DTIC Science & Technology

1976-01-01

Calibration and Poduction Runs, 606.4-1 to 606,.4-,102 A detailed index for eaoh task section precedes the respective section text. - 5- -., ~5 , 5, -𔃿.-- .55...before the mixing of Hangman Creek. Station 6. on the Hangman Creek, is located apprcximately 200 feet upstream of its confluence with the Spokane...in terms of the entire reach length considered as a mixed body of uniform quality. Therefore, the selection of reaches is also 606.1-8 tied to the

Variable partitioning of flow and sediment transfer through a large river diffluence-confluence unit across a monsoonal flood pulse

NASA Astrophysics Data System (ADS)

Hackney, C. R.; Aalto, R. E.; Darby, S. E.; Parsons, D. R.; Leyland, J.; Nicholas, A. P.; Best, J.

2016-12-01

Bifurcations represent key morphological nodes within the channel networks of anabranching and braided fluvial channels, playing an important role in controlling local bed morphology, the routing of sediment and water, and defining the stability of the downstream reaches. Herein, we detail field observations of the three-dimensional flow structure, bed morphological changes and partitioning of both flow discharge and suspended sediment through a large diffluence-confluence unit on the Mekong River, Cambodia, across a range of flow stages (from 13,500 m3 s-1 to 27,000 m3 s-1) over the monsoonal flood-pulse cycle. We show that the discharge asymmetry (a measure of the disparity between discharges distributed down the left and right branches of the bifurcation) varies with flow discharge and that the influence of upstream curvature-induced cross-stream water surface slope and bed morphological changes are first-order controls in modulating the asymmetry in bifurcation discharge. Flow discharge is shown to play a key role in defining the morphodynamics of the diffluence-confluence unit downstream of the bifurcation. Our data show that during peak flows (Q 27,000 m3 s-1), the downstream island complex acts as a net sink of suspended sediment (with 2600 kg s-1 being deposited between the diffluence and confluence), whereas during lower flows, on both the rising and falling limbs of the flood wave, the sediment balance is in quasi-equilibrium. We propose a new conceptual model of bifurcation stability that incorporates varying flood discharge and in which the long term stability of the bifurcation, as well as the larger channel planform and morphology of the diffluence-confluence unit, are controlled by the variations in flood discharge.

The influence of flow discharge variations on the morphodynamics of a diffluence-confluence unit on the Mekong River

NASA Astrophysics Data System (ADS)

Hackney, Christopher; Darby, Stephen; Parsons, Daniel; Leyland, Julian; Aalto, Rolf; Nicholas, Andrew; Best, Jim

2017-04-01

Bifurcations represent key morphological nodes within the channel networks of anabranching and braided fluvial channels, playing an important role in controlling local bed morphology, the routing of sediment and water, and defining the stability of the downstream reaches. Herein, we detail field observations of the three-dimensional flow structure, bed morphological changes and partitioning of both flow discharge and suspended sediment through a large diffluence-confluence unit on the Mekong River, Cambodia, across a range of flow stages (from 13,500 m3 s-1 to 27,000 m3 s-1) over the monsoonal flood-pulse cycle. We show that the discharge asymmetry (a measure of the disparity between discharges distributed down the left and right branches of the bifurcation) varies with flow discharge and that the influence of upstream curvature-induced cross-stream water surface slope and bed morphological changes are first-order controls in modulating the asymmetry in bifurcation discharge. Flow discharge is shown to play a key role in defining the morphodynamics of the diffluence-confluence unit downstream of the bifurcation. Our data show that during high flows (Q 27,000 m3 s-1), the downstream island complex acts as a net sink of suspended sediment (with 2600 kg s-1 being deposited between the diffluence and confluence), whereas during lower flows, on both the rising and falling limbs of the flood wave, the sediment balance is in quasi-equilibrium. We propose, therefore, that the long term stability of the bifurcation, as well as the larger channel planform and morphology of the diffluence-confluence unit, is therefore controlled by annual monsoonal flood pulses and the associated variations in discharge.

Fisetin and luteolin protect human retinal pigment epithelial cells from oxidative stress-induced cell death and regulate inflammation

PubMed Central

Hytti, Maria; Piippo, Niina; Korhonen, Eveliina; Honkakoski, Paavo; Kaarniranta, Kai; Kauppinen, Anu

2015-01-01

Degeneration of retinal pigment epithelial (RPE) cells is a clinical hallmark of age-related macular degeneration (AMD), the leading cause of blindness among aged people in the Western world. Both inflammation and oxidative stress are known to play vital roles in the development of this disease. Here, we assess the ability of fisetin and luteolin, to protect ARPE-19 cells from oxidative stress-induced cell death and to decrease intracellular inflammation. We also compare the growth and reactivity of human ARPE-19 cells in serum-free and serum-containing conditions. The absence of serum in the culture medium did not prevent ARPE-19 cells from reaching full confluency but caused an increased sensitivity to oxidative stress-induced cell death. Both fisetin and luteolin protected ARPE-19 cells from oxidative stress-induced cell death. They also significantly decreased the release of pro-inflammatory cytokines into the culture medium. The decrease in inflammation was associated with reduced activation of MAPKs and CREB, but was not linked to NF- κB or SIRT1. The ability of fisetin and luteolin to protect and repair stressed RPE cells even after the oxidative insult make them attractive in the search for treatments for AMD. PMID:26619957

Cholinergic and dopaminergic neuronal differentiation of human adipose tissue derived mesenchymal stem cells.

PubMed

Marei, Hany El Sayed; El-Gamal, Aya; Althani, Asma; Afifi, Nahla; Abd-Elmaksoud, Ahmed; Farag, Amany; Cenciarelli, Carlo; Thomas, Caceci; Anwarul, Hasan

2018-02-01

Mesenchymal stem cells (MSCs) are multipotent cells that can differentiate into various cell types such as cartilage, bone, and fat cells. Recent studies have shown that induction of MSCs in vitro by growth factors including epidermal growth factor (EGF) and fibroblast growth factor (FGF2) causes them to differentiate into neural like cells. These cultures also express ChAT, a cholinergic marker; and TH, a dopaminergic marker for neural cells. To establish a protocol with maximum differentiation potential, we examined MSCs under three experimental culture conditions using neural induction media containing FGF2, EGF, BMP-9, retinoic acid, and heparin. Adipose-derived MSCs were extracted and expanded in vitro for 3 passages after reaching >80% confluency, for a total duration of 9 days. Cells were then characterized by flow cytometry for CD markers as CD44 positive and CD45 negative. MSCs were then treated with neural induction media and were characterized by morphological changes and Q-PCR. Differentiated MSCs expressed markers for immature and mature neurons; β Tubulin III (TUBB3) and MAP2, respectively, showing the neural potential of these cells to differentiate into functional neurons. Improved protocols for MSCs induction will facilitate and ensure the reproducibility and standard production of MSCs for therapeutic applications in neurodegenerative diseases. © 2017 Wiley Periodicals, Inc.

The Planform Mobility of Large River Channel Confluences

NASA Astrophysics Data System (ADS)

Sambrook Smith, Greg; Dixon, Simon; Nicholas, Andrew; Bull, Jon; Vardy, Mark; Best, James; Goodbred, Steven; Sarker, Maminul

2017-04-01

Large river confluences are widely acknowledged as exerting a controlling influence upon both upstream and downstream morphology and thus channel planform evolution. Despite their importance, little is known concerning their longer-term evolution and planform morphodynamics, with much of the literature focusing on confluences as representing fixed, nodal points in the fluvial network. In contrast, some studies of large sand bed rivers in India and Bangladesh have shown large river confluences can be highly mobile, although the extent to which this is representative of large confluences around the world is unknown. Confluences have also been shown to generate substantial bed scours, and if the confluence location is mobile these scours could 'comb' across wide areas. This paper presents field data of large confluences morphologies in the Ganges-Brahmaputra-Meghna river basin, illustrating the spatial extent of large river bed scours and showing scour depth can extend below base level, enhancing long term preservation potential. Based on a global review of the planform of large river confluences using Landsat imagery from 1972 to 2014 this study demonstrates such scour features can be highly mobile and there is an array of confluence morphodynamic types: from freely migrating confluences, through confluences migrating on decadal timescales to fixed confluences. Based on this analysis, a conceptual model of large river confluence types is proposed, which shows large river confluences can be sites of extensive bank erosion and avulsion, creating substantial management challenges. We quantify the abundance of mobile confluence types by classifying all large confluences in both the Amazon and Ganges-Brahmaputra-Meghna basins, showing these two large rivers have contrasting confluence morphodynamics. We show large river confluences have multiple scales of planform adjustment with important implications for river management, infrastructure and interpretation of the rock record.

Multipotent Mesenchymal Stromal Stem Cell Expansion by Plating Whole Bone Marrow at a Low Cellular Density: A More Advantageous Method for Clinical Use

PubMed Central

Mareschi, Katia; Rustichelli, Deborah; Calabrese, Roberto; Gunetti, Monica; Sanavio, Fiorella; Castiglia, Sara; Risso, Alessandra; Ferrero, Ivana; Tarella, Corrado; Fagioli, Franca

2012-01-01

Mesenchymal stem cells (MSCs) are a promising source for cell therapy due to their pluripotency and immunomodulant proprieties. As the identification of “optimal” conditions is important to identify a standard procedure for clinical use. Percoll, Ficoll and whole bone marrow directly plated were tested from the same sample as separation methods. The cells were seeded at the following densities: 100 000, 10 000, 1000, 100, 10 cells/cm2. After reaching confluence, the cells were detached, pooled and re-plated at 1000, 500, 100, and 10 cells/cm2. Statistical analyses were performed. Cumulative Population Doublings (PD) did not show significant differences for the separation methods and seeding densities but only for the plating density. Some small quantity samples plated in T25 flasks at plating densities of 10 and 100 cells/cm2 did not produce any expansion. However, directly plated whole bone marrow resulted in a more advantageous method in terms of CFU-F number, cellular growth and minimal manipulation. No differences were observed in terms of gross morphology, differentiation potential or immunophenotype. These data suggest that plating whole bone marrow at a low cellular density may represent a good procedure for MSC expansion for clinical use. PMID:23715383

Induction of morphological changes in death-induced cancer cells monitored by holographic microscopy.

PubMed

El-Schich, Zahra; Mölder, Anna; Tassidis, Helena; Härkönen, Pirkko; Falck Miniotis, Maria; Gjörloff Wingren, Anette

2015-03-01

We are using the label-free technique of holographic microscopy to analyze cellular parameters including cell number, confluence, cellular volume and area directly in the cell culture environment. We show that death-induced cells can be distinguished from untreated counterparts by the use of holographic microscopy, and we demonstrate its capability for cell death assessment. Morphological analysis of two representative cell lines (L929 and DU145) was performed in the culture flasks without any prior cell detachment. The two cell lines were treated with the anti-tumour agent etoposide for 1-3days. Measurements by holographic microscopy showed significant differences in average cell number, confluence, volume and area when comparing etoposide-treated with untreated cells. The cell volume of the treated cell lines was initially increased at early time-points. By time, cells decreased in volume, especially when treated with high doses of etoposide. In conclusion, we have shown that holographic microscopy allows label-free and completely non-invasive morphological measurements of cell growth, viability and death. Future applications could include real-time monitoring of these holographic microscopy parameters in cells in response to clinically relevant compounds. Copyright © 2015 Elsevier Inc. All rights reserved.

Oxygen mapping: Probing a novel seeding strategy for bone tissue engineering.

PubMed

Westphal, Ines; Jedelhauser, Claudia; Liebsch, Gregor; Wilhelmi, Arnd; Aszodi, Attila; Schieker, Matthias

2017-04-01

Bone tissue engineering (BTE) utilizing biomaterial scaffolds and human mesenchymal stem cells (hMSCs) is a promising approach for the treatment of bone defects. The quality of engineered tissue is crucially affected by numerous parameters including cell density and the oxygen supply. In this study, a novel oxygen-imaging sensor was introduced to monitor the oxygen distribution in three dimensional (3D) scaffolds in order to analyze a new cell-seeding strategy. Immortalized hMSCs, pre-cultured in a monolayer for 30-40% or 70-80% confluence, were used to seed demineralized bone matrix (DBM) scaffolds. Real-time measurements of oxygen consumption in vitro were simultaneously performed by the novel planar sensor and a conventional needle-type sensor over 24 h. Recorded oxygen maps of the novel planar sensor revealed that scaffolds, seeded with hMSCs harvested at lower densities (30-40% confluence), exhibited rapid exponential oxygen consumption profile. In contrast, harvesting cells at higher densities (70-80% confluence) resulted in a very slow, almost linear, oxygen decrease due to gradual achieving the stationary growth phase. In conclusion, it could be shown that not only the seeding density on a scaffold, but also the cell density at the time point of harvest is of major importance for BTE. The new cell seeding strategy of harvested MSCs at low density during its log phase could be a useful strategy for an early in vivo implantation of cell-seeded scaffolds after a shorter in vitro culture period. Furthermore, the novel oxygen imaging sensor enables a continuous, two-dimensional, quick and convenient to handle oxygen mapping for the development and optimization of tissue engineered scaffolds. Biotechnol. Bioeng. 2017;114: 894-902. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Mixing zone hydrodynamics in a large confluence: a case study of the Snake and Clearwater Rivers confluence

NASA Astrophysics Data System (ADS)

Shehata, M. M.; Petrie, J.

2015-12-01

Confluences are a basic component in all fluvial systems, which are often characterized by complex flow and sediment transport patterns. Addressing confluences, however, started only recently in parallel with new advances of flow measurement tools and computational techniques. A limited number of field studies exist investigating flow hydrodynamics through confluences, particularly for large confluences with central zone widths of 100 m or greater. Previous studies have indicated that the size of the confluent rivers and the post-confluence zone may impact flow and sediment transport processes in the confluence zone, which consequently could impact the biodiversity within the river network. This study presents the results of a field study conducted at the confluence of the Snake and the Clearwater rivers near the towns of Clarkston, WA and Lewiston, ID (average width of 700 m at the confluence center). This confluence supports many different and, sometimes, conflicting purposes including commercial navigation, recreation, and fish and wildlife conservation. The confluence properties are affected by dredging operations carried out periodically to maintain the minimum water depth required for safe flow conveyance and navigation purposes. Also, a levee system was constructed on the confluence banks as an extra flood control measure. In the recent field work, an Acoustic Doppler Current Profiler was used to measure water velocity profiles at cross sections in the confluence region. Fixed and moving vessel measurements were taken at selected locations to evaluate both the spatial and temporal variation in velocity throughout the confluence. The confluence bathymetry was surveyed with a multi-beam sonar to investigate existent bed morphological elements. The results identify the velocity pattern in the mixing zone between the two rivers. The present findings are compared to previous studies on small confluences to demonstrate the influence of scale on flow processes.

Megascours: the morphodynamics of large river confluences

NASA Astrophysics Data System (ADS)

Dixon, Simon; Sambrook Smith, Greg; Nicholas, Andrew; Best, Jim; Bull, Jon; Vardy, Mark; Goodbred, Steve; Haque Sarker, Maminul

2015-04-01

River confluences are wildly acknowledged as crucial controlling influences upon upstream and downstream morphology and thus landscape evolution. Despite their importance very little is known about their evolution and morphodynamics, and there is a consensus in the literature that confluences represent fixed, nodal points in the fluvial network. Confluences have been shown to generate substantial bed scours around five times greater than mean depth. Previous research on the Ganges-Jamuna junction has shown large river confluences can be highly mobile, potentially 'combing' bed scours across a large area, although the extent to which this is representative of large confluences in general is unknown. Understanding the migration of confluences and associated scours is important for multiple applications including: designing civil engineering infrastructure (e.g. bridges, laying cable, pipelines, etc.), sequence stratigraphic interpretation for reconstruction of past environmental and sea level change, and in the hydrocarbon industry where it is crucial to discriminate autocyclic confluence scours from widespread allocyclic surfaces. Here we present a wide-ranging global review of large river confluence planforms based on analysis of Landsat imagery from 1972 through to 2014. This demonstrates there is an array of confluence morphodynamic types: from freely migrating confluences such as the Ganges-Jamuna, through confluences migrating on decadal timescales and fixed confluences. Along with data from recent geophysical field studies in the Ganges-Brahmaputra-Meghna basin we propose a conceptual model of large river confluence types and hypothesise how these influence morphodynamics and preservation of 'megascours' in the rock record. This conceptual model has implications for sequence stratigraphic models and the correct identification of surfaces related to past sea level change. We quantify the abundance of mobile confluence types by classifying all large confluences in the Amazon and Ganges-Brahmaputra-Meghna basins, showing these two basins have contrasting confluence morphodynamics. For the first time we show large river confluences have multiple scales of planform adjustment with important implications for infrastructure and interpretation of the rock record.

Autogenous bone chips: influence of a new piezoelectric device (Piezosurgery) on chip morphology, cell viability and differentiation.

PubMed

Chiriac, G; Herten, M; Schwarz, F; Rothamel, D; Becker, J

2005-09-01

The aim of the present study was to investigate the influence of a new piezoelectric device, designed for harvesting autogenous bone chips from intra-oral sites, on chip morphology, cell viability and differentiation. A total of 69 samples of cortical bone chips were randomly gained by either (1) a piezoelectric device (PS), or (2) conventional rotating drills (RD). Shape and size of the bone chips were compared by means of morphometrical analysis. Outgrowing osteoblasts were identified by means of alkaline phosphatase activity (AP), immunhistochemical staining for osteocalcin (OC) synthesis and reverse transcriptase-polymerase chain reaction phenotyping. In 88.9% of the RD and 87.9% of the PS specimens, an outgrowth of adherent cells nearby the bone chips was observed after 6-19 days. Confluence of cells was reached after 4 weeks. Positive staining for AP and OC identified the cells as osteoblasts. The morphometrical analysis revealed a statistically significant more voluminous size of the particles collected with PS than RD. Within the limits of the present study, it may be concluded that both the harvesting methods are not different from each other concerning their detrimental effect on viability and differentiation of cells growing out of autogenous bone chips derived from intra-oral cortical sites.

Epithelial cell kinase-B61: an autocrine loop modulating intestinal epithelial migration and barrier function.

PubMed

Rosenberg, I M; Göke, M; Kanai, M; Reinecker, H C; Podolsky, D K

1997-10-01

Epithelial cell kinase (Eck) is a member of a large family of receptor tyrosine kinases whose functions remain largely unknown. Expression and regulation of Eck and its cognate ligand B61 were analyzed in the human colonic adenocarcinoma cell line Caco-2. Immunocytochemical staining demonstrated coexpression of Eck and B61 in the same cells, suggestive of an autocrine loop. Eck levels were maximal in preconfluent cells. In contrast, B61 levels were barely detectable in preconfluent cells and increased progressively after the cells reached confluence. Caco-2 cells cultured in the presence of added B61 showed a significant reduction in the levels of dipeptidyl peptidase and sucrase-isomaltase mRNA, markers of Caco-2 cell differentiation. Cytokines interleukin-1beta (IL-1beta), basic fibroblast growth factor, IL-2, epidermal growth factor, and transforming growth factor-beta modulated steady-state levels of Eck and B61 mRNA and regulated Eck activation as assessed by tyrosine phosphorylation. Functionally, stimulation of Eck by B61 resulted in increased proliferation, enhanced barrier function, and enhanced restitution of injured epithelial monolayers. These results suggest that the Eck-B61 interaction, a target of regulatory peptides, plays a role in intestinal epithelial cell development, migration, and barrier function, contributing to homeostasis and preservation of continuity of the epithelial barrier.

On measuring cell confluence in phase contrast microscopy

NASA Astrophysics Data System (ADS)

Dempsey, K. P.; Richardson, J. B.; Lam, K. P.

2014-03-01

A principal focus highlighting recent advances in cell based therapies concerns the development of effective treatments for osteoarthritis. Earlier clinicaltrials have shown that 80% of patients receiving mesenchymal stem cell(MSC) based treatment have improved their quality of life by alleviating pain whilst extending the life of their natural joints. The current challenge facing researchers is to identify the biological differences between the treatments that have worked and those which have shown little improvement. One possible candidate for the difference in treatment prognosis is an examination of the proliferation of the ( type) cells as they grow. To further understanding of the proliferation and differentiation of MSC, non-invasive live cell imaging techniques have been developed which capture important cell events and dynamics in cell divisions over an extended period of time. An automated image analysis procedure capable of tracking cell confluence over time has also been implemented, providing an objective and realistic estimation of cell growth within continuous live cell cultures. The proposed algorithm accounts for the halo artefacts that occur in phase microscopy. In addition to a favourable run-time performance, the method was also validated using continuous live MSC cultures, with consistent and meaningful results.

In vitro ovine articular chondrocyte proliferation: experiments and modelling.

PubMed

Mancuso, L; Liuzzo, M I; Fadda, S; Pisu, M; Cincotti, A; Arras, M; La Nasa, G; Concas, A; Cao, G

2010-06-01

This study focuses on analysis of in vitro cultures of chondrocytes from ovine articular cartilage. Isolated cells were seeded in Petri dishes, then expanded to confluence and phenotypically characterized by flow cytometry. The sigmoidal temporal profile of total counts was obtained by classic haemocytometry and corresponding cell size distributions were measured electronically using a Coulter Counter. A mathematical model recently proposed (1) was adopted for quantitative interpretation of these experimental data. The model is based on a 1-D (that is, mass-structured), single-staged population balance approach capable of taking into account contact inhibition at confluence. The model's parameters were determined by fitting measured total cell counts and size distributions. Model reliability was verified by predicting cell proliferation counts and corresponding size distributions at culture times longer than those used when tuning the model's parameters. It was found that adoption of cell mass as the intrinsic characteristic of a growing chondrocyte population enables sigmoidal temporal profiles of total counts in the Petri dish, as well as cell size distributions at 'balanced growth', to be adequately predicted.

Assessment of selected water-quality data collected in the lower Red River (main stem) basin, Texas, 1997-98

USGS Publications Warehouse

Baldys, Stanley; Hamilton, Danna K.

2003-01-01

The Texas part of the Red River Basin has been divided into five reaches or subbasins (fig. 1) to facilitate improved planning, monitoring, geographical analysis, and dissemination of information. The U.S. Geological Survey (USGS), in cooperation with the Red River Authority of Texas, is studying the five subbasins, each for a period of about 1 year. Baldys and Phillips (1998) discuss various components and the associated scope of study of each of the five reaches. Data from the first reach studied—reach 2, the Wichita River Basin—were presented in a fact sheet by Baldys and Phillips (2000). This fact sheet presents an assessment of data collected at 11 sites during 1997–98 for reach 1—the lower Red River (main stem) Basin from the confluence of Cache Creek downstream to the Texas-Arkansas State Line (fig. 1).

Floods in Starkweather Creek basin, Madison, Wisconsin

USGS Publications Warehouse

Lawrence, Carl L.; Holmstrom, Barry K.

1972-01-01

The reaches evaluated are (1) Starkweather Creek and West Branch Starkweather Creek, for a distance of 6.0 river miles from the mouth at Lake Monona upstream to the U.S. Highway 51 crossing north of Truax Field; and (2) East Branch Starkweather Creek (2.8 river miles), from its confluence with the West Branch near Milwaukee Street upstream to a point near the Interstate Highway 90-94 crossing.

Effect of Levonorgestrel (NORPLANT) on the Immune Regulation of Bone Morphogenesis in Calvarial Cultures from the Laboratory Mouse (Mus muscularis).

DTIC Science & Technology

1995-10-01

continually releases a synthetic progestin, levonorgestrel , for five years. In order to assess the impact of levonorgestrel on bone cells, murine calvarial...cell cultures were harvested, grown to confluence and treated with levonorgestrel , progesterone and estrogen. The majority of the cells grown in these

NaK-ATPase pump sites in cultured bovine corneal endothelium of varying cell density at confluence.

PubMed

Crawford, K M; Ernst, S A; Meyer, R F; MacCallum, D K

1995-06-01

The driving force for ion and water flow necessary for efficient deturgesence of the corneal stroma resides in the ouabain-sensitive sodium (Na) pump of corneal endothelial cells. Using a cell culture model of corneal endothelial cell hypertrophy, the authors examined the expression of Na pumps at the cell surface to see how this central element of the endothelial pump changed as corneal endothelial cell density decreased to a level associated with corneal decompensation in vivo. 3H-ouabain binding to NaK-ATPase at saturating conditions was used to quantitate the number of Na pump sites on cultured bovine corneal endothelial cells as the confluent density decreased from approximately 2750 cells/mm2 to approximately 275 cells/mm2. The mean number of Na pump sites per cell at confluence (1.92 +/- 0.07 x 10(6)) did not change as the cell density decreased 2.7-fold from 2763 cells/mm2 to 1000 cells/mm2. However, pump site expression doubled to approximately 4 x 10(6) sites/cell as the cell density decreased from 1000 cells/mm2 to 275 cells/mm2. Despite the incremental increase in Na pump site expression that occurred as the cells hypertrophied below a density of 1000/mm2 to achieve confluence, this increase was insufficient to prevent a decrease in Na pump site density of the intact monolayer, expressed as pump sites/mm2. The confluent cell density of cultured bovine corneal endothelial cells can be varied from that found in the normal native cornea to that associated with corneal decompensation. In confluent cultures with cell densities ranging from 2750 cells/mm2 to 1000 cells/mm2, the number of pump sites per cell remains relatively unchanged. Below cell densities of 1000 cells/mm2, the number of pump sites per cell progressively increases. The increased Na pump site abundance in markedly hypertrophied endothelial cells cannot adequately compensate for the progressive reduction in the number of transporting cells per unit area within the intact monolayer. Even when considered with the decrease in the size of the paracellular ion conductive pathway that is a consequence of progressive endothelial hypertrophy, the overall pumping capacity of the intact endothelial monolayer declines.

Spatial and seasonal variability of base flow in the Verde Valley, central Arizona, 2007 and 2011

USGS Publications Warehouse

Garner, Bradley D.; Bills, Donald J.

2012-01-01

Synoptic base-flow surveys were conducted on streams in the Verde Valley, central Arizona, in June 2007 and February 2011 by the U.S. Geological Survey (USGS), in cooperation with the Verde River Basin Partnership, the Town of Clarkdale, and Yavapai County. These surveys, also known as seepage runs, measured streamflow under base-flow conditions at many locations over a short period of time. Surveys were conducted on a segment of the Verde River that flows through the Verde Valley, between USGS streamflow-gaging stations 09504000 and 09506000, a distance of 51 river miles. Data from the surveys were used to investigate the dominant controls on Verde River base flow, spatial variability in gaining and losing reaches, and the effects that human alterations have on base flow in the surface-water system. The most prominent human alterations in the Verde Valley are dozens of surface-water diversions from streams, including gravity-fed ditch diversions along the Verde River.Base flow that entered the Verde River from the tributary streams of Oak Creek, Beaver Creek, and West Clear Creek was found to be a major source of base flow in the Verde River. Groundwater discharge directly into the Verde River near these three confluences also was an important contributor of base flow to the Verde River, particularly near the confluence with Beaver Creek. An examination of individual reaches of the Verde River in the Verde Valley found three reaches (largely unaffected by ditch diversions) exhibiting a similar pattern: a small net groundwater discharge in February 2011 (12 cubic feet per second or less) and a small net streamflow loss in June 2007 (11 cubic feet per second or less). Two reaches heavily affected by ditch diversions were difficult to interpret because of the large number of confounding human factors. Possible lower and upper bounds of net groundwater flux were calculated for all reaches, including those heavily affected by ditches.

Techniques for studying the effects of microgravity on model particle/cell systems

NASA Technical Reports Server (NTRS)

Young, Ronald B.

1988-01-01

To study the direct effects of a low gravity environment on skeletal and cardiac muscle cells, experiments were initiated to determine whether skeletal and/or cardiac muscule cells would grow within the lumen of XM-80 hollow fibers (i.d. = 0.5 mm). Cells were prepared from skeletal or cardiac muscle tissue of 12 day embryos and were cultured for up to 7 days in the hollow fiber environment. Light microscopy revealed that cells proliferated to confluency over this period of time and fusion was apparent in the skeletal muscle cells. Once it was verified that cells would grow to confluency, additional XM-80 fibers containing cells were placed in a Clinostat in the horizontal position at 100 rpm. Fibers were stretched by a built-in spring mechanism to hold the fiber tightly at the center of rotation. Under these conditions, the gravity vector approaches zero and the cells are in an environment that simulates microgravity. Examination of skeletal muscle cells by electron microscopy revealed that myoblast fusion and myofibril accumulation were extensive. Although data obtained thus far are preliminary, they suggest that myofibril organization in chicken skeletal muscle cultures is somewhat more poorly defined in Clinostat rotated cultures than in controls that were not subjected to Clinostat conditions.

Flood-inundation maps for Indian Creek and Tomahawk Creek, Johnson County, Kansas, 2014

USGS Publications Warehouse

Peters, Arin J.; Studley, Seth E.

2016-01-25

Digital flood-inundation maps for a 6.4-mile upper reach of Indian Creek from College Boulevard to the confluence with Tomahawk Creek, a 3.9-mile reach of Tomahawk Creek from 127th Street to the confluence with Indian Creek, and a 1.9-mile lower reach of Indian Creek from the confluence with Tomahawk Creek to just beyond the Kansas/Missouri border at State Line Road in Johnson County, Kansas, were created by the U.S. Geological Survey in cooperation with the city of Overland Park, Kansas. The flood-inundation maps, which can be accessed through the U.S. Geological Survey Flood Inundation Mapping Science Web site at http://water.usgs.gov/osw/flood_inundation/, depict estimates of the areal extent and depth of flooding corresponding to selected water levels (stages) at the U.S. Geological Survey streamgages on Indian Creek at Overland Park, Kansas; Indian Creek at State Line Road, Leawood, Kansas; and Tomahawk Creek near Overland Park, Kansas. Near real time stages at these streamgages may be obtained on the Web from the U.S. Geological Survey National Water Information System at http://waterdata.usgs.gov/nwis or the National Weather Service Advanced Hydrologic Prediction Service at http://water.weather.gov/ahps/, which also forecasts flood hydrographs at these sites.Flood profiles were computed for the stream reaches by means of a one-dimensional step-backwater model. The model was calibrated for each reach by using the most current stage-discharge relations at the streamgages. The hydraulic models were then used to determine 15 water-surface profiles for Indian Creek at Overland Park, Kansas; 17 water-surface profiles for Indian Creek at State Line Road, Leawood, Kansas; and 14 water-surface profiles for Tomahawk Creek near Overland Park, Kansas, for flood stages at 1-foot intervals referenced to the streamgage datum and ranging from bankfull to the next interval above the 0.2-percent annual exceedance probability flood level (500-year recurrence interval). The simulated water-surface profiles were then combined in a geographic information system with a digital elevation model derived from light detection and ranging data (having a 0.429-foot vertical and 0.228-foot horizontal accuracy) to delineate the area flooded at each water level.The availability of these maps, along with Web information regarding current stage from the U.S. Geological Survey streamgages and forecasted high-flow stages from the National Weather Service, will provide emergency management personnel and residents with information that is critical for flood response activities such as evacuations, road closures, and postflood recovery efforts.

Diurnal cycles control the fate of contaminants at an Andean river confluence impacted by legacy mining

NASA Astrophysics Data System (ADS)

Pasten, P.; Guerra, P. A.; Simonson, K.; Bonilla, C.; Pizarro, G. E.; Escauriaza, C. R.; González, C.

2014-12-01

The importance of hydrologic-geochemical interactions in arid environments is a controlling factor in quality and quantity of water available for human consumption and agriculture. When acid drainage affects these watersheds, water quality is gravely degraded. Despite its effect on watersheds, the relationship between time changes in hydrological variables and water quality in arid regions has not been studied thoroughly. Temporal variations in acid drainage can control when the transport of toxic elements is increased. We performed field work at the Azufre River (pH 2, E.C~10.9 mS/cm) and Caracarani River (pH 8.7, E.C~1.2 mS/cm) confluence, located in the Northern Chilean Altiplano (at 4000 m asl). We registered stream flowrates (total flowrate~430 L/s), temperature and electric conductivity (E.C) hourly using in-stream data loggers during one year. We also measured turbidity and pH during one field survey at different distances from the junction, as a proxy of the formation of iron-aluminum particles that cycle trace elements in these environments. We found turbidity-pH diurnal cycles were caused by upstream hourly changes in upstream flowrate: when the Caracarani River flowrate reached its daily peak, particle formation occurred, while the dissolution of particles occurred when the Azufre River reached its maximum value. This last process occurred due to upstream freeze-thaw cycles. This study shows how the dynamics of natural confluences determines chemical transport. The formation of particles enriched in toxic elements can promote settling as a natural attenuation process, while their dissolution will produce their release and transport long distances downstream. It is important to consider time as an important variable in water quality monitoring and in water management infrastructure where pulses of contamination can have potentially negative effects in its use. Acknowledgements: Funding was provided by "Proyecto Fondecyt 1130936" and "CONICYT/FONDAP 15110020".

Sand deposition in the Colorado River in the Grand Canyon from flooding of the Little Colorado River

USGS Publications Warehouse

Wiele, S.M.; Graf, J.B.; Smith, J.D.

1996-01-01

Methods for computing the volume of sand deposited in the Colorado River in Grand Canyon National Park by floods in major tributaries and for determining redistribution of that sand by main-channel flows are required for successful management of sand-dependent riparian resources. We have derived flow, sediment transport, and bed evolution models based on a gridded topography developed from measured channel topography and used these models to compute deposition in a short reach of the river just downstream from the Little Colorado River, the largest tributary in the park. Model computations of deposition from a Little Colorado River flood in January 1993 were compared to bed changes measured at 15 cross sections. The total difference between changes in cross-sectional area due to deposition computed by the model and the measured changes was 6%. A wide reach with large areas of recirculating flow and large depressions in the main channel accumulated the most sand, whereas a reach with similar planimetric area but a long, narrow shape and relatively small areas of recirculating flow and small depressions in the main channel accumulated only about a seventh as much sand. About 32% of the total deposition was in recirculation zones, 65% was in the main channel, and 3% was deposited along the channel margin away from the recirculation zone. Overall, about 15% of the total input of sand from this Little Colorado River flood was deposited in the first 3 km below the confluence, suggesting that deposition of the flood-derived material extended for only several tens of kilometers downstream from the confluence.

Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl.

PubMed

Stamm, Anne; Strauß, Sarah; Vogt, Peter; Scheper, Thomas; Pepelanova, Iliyana

2018-04-07

AmbLOXe is a lipoxygenase, which is up-regulated during limb-redevelopment in the Mexican axolotl, Ambystoma mexicanum, an animal with remarkable regeneration capacity. Previous studies have shown that mammalian cells transformed with the gene of this epidermal lipoxygenase display faster migration and wound closure rate during in vitro wound healing experiments. In this study, the gene of AmbLOXe was codon-optimized for expression in Escherichia coli and was produced in the insoluble fraction as protein aggregates. These inclusion bodies or nanopills were shown to be reservoirs containing functional protein during in vitro wound healing assays. For this purpose, functional inclusion bodies were used to coat cell culture surfaces prior cell seeding or were added directly to the medium after cells reached confluence. In both scenarios, AmbLOXe inclusion bodies led to faster migration rate and wound closure, in comparison to controls containing either no AmbLOXe or GFP inclusion bodies. Our results demonstrate that AmbLOXe inclusion bodies are functional and may serve as stable reservoirs of this enzyme. Nevertheless, further studies with soluble enzyme are also necessary in order to start elucidating the exact molecular substrates of AmbLOXe and the biochemical pathways involved in the wound healing effect.

Inhibition of protein phosphorylation in MIA pancreatic cancer cells: Confluence of metabolic and signaling pathways

PubMed Central

Zhang, Hengwei; Cao, Rui; Lee, Wai-Nang Paul; Deng, Caishu; Zhao, Yingchun; Lappe, Joan; Recker, Robert; Yen, Yun; Wang, Qi; Tsai, Ming-Ying; Go, Vay Liang; Xiao, Gary Guishan

2010-01-01

Oxythiamine (OT), a transketolase inhibitor, is known to inhibit pancreatic cancer cell proliferation. In this study, we investigated the effect of inhibition of the transketolase pathway on signaling pathways in MIA PaCa cancer cells using in-house proteomic techniques. We hypothesized that OT alter protein phosphorylation thus affecting cell cycle arrest and cell proliferation. MIA PaCa-2 cells were cultured in media containing an algal 15N amino acid mixture at 50% enrichment, with and without OT, to determine protein expression and synthesis. Analysis of cell lysates using two-dimensional gel electrophoresis matrix assisted laser desorption and ionization time-of-flight and time-of-flight mass spectrometry (2-DE-MALDI-TOF/TOF MS) identified 12 phosphor proteins that were significantly suppressed by OT treatment. Many of these proteins are involved in regulation of cycle activities and apoptosis. Among the proteins identified, expression of the phosphor heat shock protein 27 (Hsp27) was dramatically inhibited by OT treatment while the level of its total protein remained unchanged. Hsp27 expression and phoshporylation is known to be associated with drug resistance and cancer cell survival. The changes in phosphorylation of key proteins of cancer proliferation and survival suggest that protein phosphorylation is the confluence of the effects of OT on metabolic and signaling pathways. PMID:20035555

Comparison of the canine corneal epithelial cell sheets cultivated from limbal stem cells on canine amniotic membrane, atelocollagen gel, and temperature-responsive culture dish.

PubMed

Nam, Eunryel; Fujita, Naoki; Morita, Maresuke; Tsuzuki, Keiko; Lin, Hsing Yi; Chung, Cheng Shu; Nakagawa, Takayuki; Nishimura, Ryohei

2015-07-01

The current study compared canine corneal epithelial cell sheets cultivated from limbal stem cells on amniotic membrane, atelocollagen gel, and temperature-responsive culture dish. We collected limbal epithelial cells from the intact eyes of beagles and cultivated the cells on denuded canine amniotic membranes, temperature-responsive cell culture labware, and collagen gel with 3T3 feeder cells. Immunofluorescence staining for Ki-67 was used to analyze the capacity of cell proliferation in the sheets. Immunofluorescence staining was also performed for the corneal epithelium-specific marker cytokeratin 3 and putative stem cell markers ABCG2 and p63. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to detect ABCG2 and p63. The growth rates of the cultivated cells, or the times it took them to reach confluency, were different for the three scaffolds. The cultivated sheet on the temperature-responsive dish consisted of 2-3 layers, while those on the collagen gel and on the amniotic membrane consisted of 5-8 layers. The basal layer cells grown on all three scaffolds expressed putative stem cell markers. In real-time RT-PCR analysis, the highest level of p63 was observed in the sheets grown on collagen gel. In this study, the cells cultured on the collagen gel demonstrated a capacity for cell proliferation, and the expressions of stem cells in the sheets suggested that collagen gel is the most suitable carrier for clinical use. © 2014 American College of Veterinary Ophthalmologists.

Are endothelial cell bioeffects from acoustic droplet vaporization proximity dependent?

NASA Astrophysics Data System (ADS)

Seda, Robinson; Li, David; Fowlkes, J. Brian; Bull, Joseph

2013-11-01

Acoustic droplet vaporization (ADV) produces gas microbubbles that provide a means of selective occlusion in gas embolotherapy. Vaporization and subsequent occlusion occur inside blood vessels supplying the targeted tissue, such as tumors. Theoretical and computational studies showed that ADV within a vessel can impart high fluid mechanical stresses on the vessel wall. Previous in vitro studies have demonstrated that vaporization at an endothelial layer may affect cell attachment and viability. The current study is aimed at investigating the role of vaporization distance away from the endothelial layer. HUVECs were cultured in OptiCell™ chambers until reaching confluence. Dodecafluoropentane microdroplets were added, attaining a 10:1 droplet to cell ratio. A single ultrasound pulse (7.5 MHz) consisting of 16 cycles (~ 2 μs) and a 5 MPa peak rarefactional pressure was used to produce ADV while varying the vaporization distance from the endothelial layer (0 μm, 500 μm, 1000 μm). Results indicated that cell attachment and viability was significantly different if the distance was 0 μm (at the endothelial layer). Other distances were not significantly different from the control. ADV will significantly affect the endothelium if droplets are in direct contact with the cells. Droplet concentration and flow conditions inside blood vessels may play an important role. This work was supported by NIH grant R01EB006476.

77 FR 76420 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2012-12-28

... Fort Gay, confluence. Unincorporated Areas of Wayne County. At the Tug Fork +575 confluence. Mill Creek (backwater effects from From the Tug Fork +575 Town of Fort Gay. Tug Fork). confluence to approximately 1.1 miles upstream of the Tug Fork confluence. Tug Fork At the Big Sandy River +575 Town of Fort Gay...

76 FR 70397 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-14

... the Ohio River +549 +550 Town of Fort Gay, Unincorporated Areas of Wayne County. confluence. At the... of Fort Gay. Fork). confluence to approximately 1.1 miles upstream of the Tug Fork confluence. Tug Fork At the Big Sandy River +576 +575 Town of Fort Gay. confluence. Approximately 0.5 mile +577 +575...

Formation of fluvial knickzones in Japanese mountainous areas: A spatial analysis using GIS and DEMs

NASA Astrophysics Data System (ADS)

Hayakawa, Y. S.; Oguchi, T.

2006-12-01

Fluvial knickzones are the elements of bedrock rivers that can enhance stream erosion into bedrock, and they can be key morphologies highlighting interactions among earth surface processes such as erosion, tectonics, and volcanism. This study examines the longitudinal profiles of Japanese mountain rivers to illustrate the distribution of knickzones and discusses their role in the landscape development. Using 50-m DEMs, knickzones were extracted based on a quantitative criterion, and 5,753 knickzones were identified in the rivers of ca. 65,000 km long. The location of the knickzones was then examined along with other GIS data including topography, geology and precipitation. Overall, topographical conditions have the strongest influences on knickzone abundance, and upstream steep reaches of the rivers are more favorable for knickzone existence. The knickzone abundance for each rock type is also controlled by stream gradients, and lighologic boundaries do not show significant correlations with the knickzone locations. The controls of lithologic substrate on the knickzone locations are therefore limited. The abundant knickzones in steep river reaches indicate a hydraulic origin of knickzones, where stream erosions have enough strength in shaping the bedrock. Moreover, the knickzones are frequently observed in reaches slightly upstream from the major confluences at which stream discharge abruptly increases, indicating that the hydraulic anomalies of water flows at the confluences can cause knickzones which may later migrate upstream. The other possible causes of knickzone initiation including volcanic, tectonic and climatic effects are also suggested. The abundant knickzones in Japanese mountain rivers, resulted from the interactions among surface processes, suggest that river morphology modeling needs to consider the initiation and development of knickzones. tokyo.ac.jp/~hayakawa/

Nature of distribution of mercury in the sediments of the river Yamuna (tributary of the Ganges), India.

PubMed

Subramanian, V; Madhavan, N; Saxena, Rajinder; Lundin, Lars-Christer

2003-06-01

Suspended Particulate Matter (SPM), surface (bed sediments) and short length cores of sediments collected from the largest tributary of the river Ganges, namely the river Yamuna, were analysed for total mercury as well as its fractionation in various size and chemical sites in the sediments following standard procedures. Also, attempts were made to determine the vertical distribution in sediments in relation to the recent timescale of a few decades. Our observations indicate that the SPM in general showed higher levels of total mercury compared to the surface sediments while at places the enhancement could be by a factor of 10, say around 25 microg g(-1) in the downstream region that integrates the industrial midstream and agricultural downstream terrain near its confluence with the Ganges. Surface sediments in the upstream direction near the Himalayan foothills and SPM in the lower reaches showed significant high Index of Geoaccumulation (Igeo) as defined by Müller. Size fractionation studies indicate that the finer fraction preferentially showed higher levels of mercury while in the lower reaches of the river, the total mercury is equitably distributed among all size fractions. The proportion of the residual fraction of mercury in relation to mobile fractions, in general decreases downstream towards its confluence with the Ganges river. In sediment cores, the vertical distribution show systematic peaks of mercury indicating that addition of this toxic metal to the aquatic system is in direct proportion to the increase in various types of human activities such as thermal power plants, land use changes (urbanisation) in the midstream region and intensive fertiliser application in lower reaches of this vast river basin.

Retrieval of a periodontally compromised tooth by allogeneic grafting of mesenchymal stem cells from dental pulp: A case report.

PubMed

Hernández-Monjaraz, Beatriz; Santiago-Osorio, Edelmiro; Ledesma-Martínez, Edgar; Alcauter-Zavala, Andrés; Mendoza-Núñez, Víctor Manuel

2018-01-01

Objective To report a case of successful allogeneic grafting of mesenchymal dental pulp stem cells (DPSCs) as preliminary findings in a patient with periodontal disease enrolled into clinical trial ISRCTN12831118. Methods Mesenchymal stem cells from the dental pulp of a deciduous tooth from a 7-year-old donor were separated from the pulp chamber and processed via enzymatic digestion and centrifugation. DPSCs were passaged and cultured on a 35 × 13 mm culture dish in minimum essential medium-alpha, without supplementation. After reaching 80% confluency, 5 x 10 6 allogeneic DPSCs in 250 µl phosphate buffered saline were seeded onto a dry scaffold of lyophilized collagen-polyvinylpyrrolidone sponge placed in the left lower premolar area of a 61-year-old patient with periodontal disease. Surgical access to the lower premolar area was achieved using the flap technique. Results At 3 and 6 months following allogeneic graft, the patient showed no sign of rejection and exhibited decreases in tooth mobility, periodontal pocket depth and bone defect area. Bone mineral density had increased at the graft site. Conclusions Regenerative periodontal therapy using DPSCs of allogeneic origin may be a promising treatment for periodontal disease-induced bone defects.

Hydraulic Conductivity of Smooth Muscle Cell-Initiated Arterial Cocultures

PubMed Central

Mathura, Rishi A.; Russell-Puleri, Sparkle; Cancel, Limary M.; Tarbell, John M.

2015-01-01

The purpose of the study was to examine the effects of arterial coculture conditions on the transport properties of several in vitro endothelial cell (EC) – smooth muscle cell (SMC) – porous filter constructs in which SMC were grown to confluence first and then EC were inoculated. This order of culturing simulates the environment of a blood vessel wall after endothelial layer damage due to stenting, vascular grafting or other vascular wall insult. For all coculture configurations examined, we observed that hydraulic conductivity (Lp) values were significantly higher than predicted by a resistances-in-series (RIS) model accounting for the Lp of EC and SMC measured separately. The greatest increases were observed when EC were plated directly on top of a confluent SMC layer without an intervening filter, presumably mediated by direct EC – SMC contacts that were observed under confocal microscopy. The results are the opposite of a previous study that showed Lp was significantly reduced compared to an RIS model when EC were grown to confluency first. The physiological, pathophysiological and tissue engineering implications of these results are discussed. PMID:26265460

Hydraulic Conductivity of Smooth Muscle Cell-Initiated Arterial Cocultures.

PubMed

Mathura, Rishi A; Russell-Puleri, Sparkle; Cancel, Limary M; Tarbell, John M

2016-05-01

The purpose of the study was to examine the effects of arterial coculture conditions on the transport properties of several in vitro endothelial cell (EC)-smooth muscle cell (SMC)-porous filter constructs in which SMC were grown to confluence first and then EC were inoculated. This order of culturing simulates the environment of a blood vessel wall after endothelial layer damage due to stenting, vascular grafting or other vascular wall insult. For all coculture configurations examined, we observed that hydraulic conductivity (L(p)) values were significantly higher than predicted by a resistances-in-series (RIS) model accounting for the L(p) of EC and SMC measured separately. The greatest increases were observed when EC were plated directly on top of a confluent SMC layer without an intervening filter, presumably mediated by direct EC-SMC contacts that were observed under confocal microscopy. The results are the opposite of a previous study that showed L(p) was significantly reduced compared to an RIS model when EC were grown to confluency first. The physiological, pathophysiological and tissue engineering implications of these results are discussed.

A field study of the confluence between Negro and Solimões Rivers. Part 1: Hydrodynamics and sediment transport

NASA Astrophysics Data System (ADS)

Gualtieri, Carlo; Filizola, Naziano; de Oliveira, Marco; Santos, Andrè Martinelli; Ianniruberto, Marco

2018-01-01

Confluences are a common feature of riverine systems, where are located converging flow streamlines and potential mixing of separate flows. The confluence of the Negro and Solimões Rivers ranks among the largest on Earth and its study may provide some general insights into large confluence dynamics and processes. An investigation was recently conducted about that confluence in both low and high-flow conditions using acoustic Doppler velocity profiling (ADCP), water quality sampling and high-resolution seismic data. First, the study gained insights into the characterization of the basic hydrodynamics parameters about the confluence as well as of those affecting sediments transport. Second, the analysis of the results showed that common hydrodynamic features noted in previous confluence studies were herein observed. Finally, some differences between low-flow and relatively high-flow conditions about the transfer of momentum from the Solimões to the Negro side of the Amazon Channel were identified.

Astrocytes from adult Wistar rats aged in vitro show changes in glial functions.

PubMed

Souza, Débora Guerini; Bellaver, Bruna; Raupp, Gustavo Santos; Souza, Diogo Onofre; Quincozes-Santos, André

2015-11-01

Astrocytes, the most versatile cells of the central nervous system, play an important role in the regulation of neurotransmitter homeostasis, energy metabolism, antioxidant defenses and the anti-inflammatory response. Recently, our group characterized cortical astrocyte cultures from adult Wistar rats. In line with that work, we studied glial function using an experimental in vitro model of aging astrocytes (30 days in vitro after reaching confluence) from newborn (NB), adult (AD) and aged (AG) Wistar rats. We evaluated metabolic parameters, such as the glucose uptake, glutamine synthetase (GS) activity, and glutathione (GSH) content, as well as the GFAP, GLUT-1 and xCT expression. AD and AG astrocytes take up less glucose than NB astrocytes and had decreased GLUT1 expression levels. Furthermore, AD and AG astrocytes exhibited decreased GS activity compared to NB cells. Simultaneously, AD and AG astrocytes showed an increase in GSH levels, along with an increase in xCT expression. NB, AD and AG astrocytes presented similar morphology; however, differences in GFAP levels were observed. Taken together, these results improve the knowledge of cerebral senescence and represent an innovative tool for brain studies of aging. Copyright © 2015 Elsevier Ltd. All rights reserved.

75 FR 29296 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-25

... the confluence with Marble Creek. Kentucky River Tributary 2 From the confluence with the None +573... the Kentucky River. Marble Creek (Backwater effects from From the confluence with the None +580...

An Assessment of Stream Confluence Flow Dynamics using Large Scale Particle Image Velocimetry Captured from Unmanned Aerial Systems

NASA Astrophysics Data System (ADS)

Lewis, Q. W.; Rhoads, B. L.

2017-12-01

The merging of rivers at confluences results in complex three-dimensional flow patterns that influence sediment transport, bed morphology, downstream mixing, and physical habitat conditions. The capacity to characterize comprehensively flow at confluences using traditional sensors, such as acoustic Doppler velocimeters and profiles, is limited by the restricted spatial resolution of these sensors and difficulties in measuring velocities simultaneously at many locations within a confluence. This study assesses two-dimensional surficial patterns of flow structure at a small stream confluence in Illinois, USA, using large scale particle image velocimetry (LSPIV) derived from videos captured by unmanned aerial systems (UAS). The method captures surface velocity patterns at high spatial and temporal resolution over multiple scales, ranging from the entire confluence to details of flow within the confluence mixing interface. Flow patterns at high momentum ratio are compared to flow patterns when the two incoming flows have nearly equal momentum flux. Mean surface flow patterns during the two types of events provide details on mean patterns of surface flow in different hydrodynamic regions of the confluence and on changes in these patterns with changing momentum flux ratio. LSPIV data derived from the highest resolution imagery also reveal general characteristics of large-scale vortices that form along the shear layer between the flows during the high-momentum ratio event. The results indicate that the use of LSPIV and UAS is well-suited for capturing in detail mean surface patterns of flow at small confluences, but that characterization of evolving turbulent structures is limited by scale considerations related to structure size, image resolution, and camera instability. Complementary methods, including camera platforms mounted at fixed positions close to the water surface, provide opportunities to accurately characterize evolving turbulent flow structures in confluences.

33 CFR 165.T11-0511 - Safety Zone; Big Sioux River from the Military Road Bridge North Sioux City to the confluence of...

Code of Federal Regulations, 2011 CFR

2011-07-01

... the Military Road Bridge North Sioux City to the confluence of the Missouri River, SD. 165.T11-0511... River from the Military Road Bridge North Sioux City to the confluence of the Missouri River, SD. (a... Bridge, North Sioux City, SD at 42.52 degrees North, 096.48 West longitude to the confluence of the...

Pollution of the River Niger and its main tributaries

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nwokedi, G.I.C.; Obodo, G.A.

1993-08-01

The River Niger system, with a length of about 4200 kilometers, and a discharge volume of 190 cubic kilometers, per year is the third largest river in Africa, and the largest in West Africa. It serves as an important waterway for the transportation of goods and provides rich agricultural flood basins for the cultivation of food and vegetables. Also it is a major source of animal proteins in form of fishes, snails and other aquatics. Above all the River and its tributaries represent the main source of domestic water supply for the rural communities, and water for irrigation. Therefore theremore » is a need to establish the nature and present levels of pollutants in the river, and the contribution made by the tributaries to the gross pollution level. A number of studies have been reported. Martins reported on the geochemistry of the River Niger while Nriagu; Livingstone; and Imevbore provided some chemical data on the upper reaches around and above its confluence with River Benue at Lokoja. Ajayi and Osibanjo reported on the chemical properties of some tributaries above the confluence of the Niger and the Benue. So far no work has been reported on the lower reaches of the Niger where contributions of the Benue and other major tributaries are significant, and where there are large settlements on its banks and the banks of the tributaries. This work aims at establishing base-line levels of the various pollutants and their sources. 12 refs., 1 fig., 2 tabs.« less

Spatial and temporal patterns of debris flow deposition in the Oregon Coast Range, USA

USGS Publications Warehouse

May, Christine L.; Gresswell, Robert E.

2004-01-01

Patterns of debris-flow occurrence were investigated in 125 headwater basins in the Oregon Coast Range. Time since the previous debris-flows was established using dendrochronology, and recurrence interval estimates ranged from 98 to 357 years. Tributary basins with larger drainage areas had a greater abundance of potential landslide source areas and a greater frequency of scouring events compared to smaller basins. The flux rate of material delivered to the confluence with a larger river influenced the development of small-scale debris-flow fans. Fans at the mouths of tributary basins with smaller drainage areas had a higher likelihood of being eroded by the mainstem river in the interval between debris-flows, compared to bigger basins that had larger, more persistent fans. Valley floor width of the receiving channel also influenced fan development because it limited the space available to accommodate fan formation. Of 63 recent debris-flows, 52% delivered sediment and wood directly to the mainstem river, 30% were deposited on an existing fan before reaching the mainstem, and 18% were deposited within the confines of the tributary valley before reaching the confluence. Spatial variation in the location of past and present depositional surfaces indicated that sequential debris-flow deposits did not consistently form in the same place. Instead of being spatially deterministic, results of this study suggest that temporally variable and stochastic factors may be important for predicting the runout length of debris-flows.

2. CONFLUENCE POOL, DETAIL OF TUNNEL PORTAL WITH WATER ENTERING ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

2. CONFLUENCE POOL, DETAIL OF TUNNEL PORTAL WITH WATER ENTERING FROM SANTA ANA RIVER. VIEW TO NORTHEAST. - Santa Ana River Hydroelectric System, Bear Creek Diversion Dam & Confluence Pool, Redlands, San Bernardino County, CA

A field study of the confluence between Negro and Solimões Rivers. Part 2: Bed morphology and stratigraphy

NASA Astrophysics Data System (ADS)

Ianniruberto, Marco; Trevethan, Mark; Pinheiro, Arthur; Andrade, Joao Fernando; Dantas, Elton; Filizola, Naziano; Santos, André; Gualtieri, Carlo

2018-01-01

The confluence of the Negro and Solimões Rivers is an interesting study area under several points of view: it represents the second largest river confluence of the Amazon Basin; the rivers are characterized by very distinct hydrologic behaviour; and it is situated in a peculiar tectonic setting. A field investigation was undertaken to study the characteristics of this confluence, aiming to better understand the bed morphology and stratigraphy resulting from the complex interaction of geological setting, hydrodynamics, and sediment load. Two field campaigns were carried out, during low- and high-flow conditions, using high-resolution seismic, echosounding, and acoustic Doppler current profiling. A third campaign was carried out just in a limited area of the confluence, with a multi-beam echosounder. The results of these surveys provided a more detailed view of the geology, morphology and sediment distribution about the confluence.

Simulation of bone resorption-repair coupling in vitro.

PubMed

Jones, S J; Gray, C; Boyde, A

1994-10-01

In the normal adult human skeleton, new bone formation by osteoblasts restores the contours of bone surfaces following osteoclastic bone resorption, but the evidence for resorption-repair coupling remains circumstantial. To investigate whether sites of prior resorption, more than the surrounding unresorbed surface, attract osteoblasts or stimulate them to proliferate or make new matrix, we developed a simple in vitro system in which resorption-repair coupling occurs. Resorption pits were produced in mammalian dentine or bone slabs by culturing chick bone-derived cells on them for 2-3 days. The chick cells were swept off and the substrata reseeded with rat calvarial osteoblastic cells, which make bone nodules in vitro, for periods of up to 8 weeks. Cell positions and new bone formation were investigated by ordinary light microscopy, fluorescence and reflection confocal laser microscopy, and SEM, in stained and unstained samples. There was no evidence that the osteoblasts were especially attracted to, or influenced by, the sites of resorption in dentine or bone before cell confluence was reached. Bone formation was identified by light microscopy by the accumulation of matrix, staining with alizarin and calcein and by von Kossa's method, and confirmed by scanning electron microscopy (SEM) by using backscattered electron (BSE) and transmitted electron imaging of unembedded samples and BSE imaging of micro-milled embedded material. These new bone patches were located initially in the resorption pits. The model in vitro system may throw new light on the factors that control resorption-repair coupling in the mineralised tissues in vivo.

Mechanisms of Ionizing Radiation-Induced Cell Death in Primary Lung Cells

DTIC Science & Technology

2013-03-05

haemocytometer, and plated on 60 mm dishes coated with 0.5% gelatin (modification to allow attachment of PAEC). The cells were incubated for 14 days...we performed delayed plating where PAEC were first cultured to 70-90% confluence, exposed to varying doses of X-rays, incubated, and seeded then for...calculated as a function of plating efficiency of non-irradiated controls. The plating efficiency (PE) was defined as the percentage of the number

Carcinogens induce loss of the primary cilium in human renal proximal tubular epithelial cells independently of effects on the cell cycle

PubMed Central

Radford, Robert; Slattery, Craig; Jennings, Paul; Blacque, Oliver; Pfaller, Walter; Gmuender, Hans; Van Delft, Joost; Ryan, Michael P.

2012-01-01

The primary cilium is an immotile sensory and signaling organelle found on the majority of mammalian cell types. Of the multitude of roles that the primary cilium performs, perhaps some of the most important include maintenance of differentiation, quiescence, and cellular polarity. Given that the progression of cancer requires disruption of all of these processes, we have investigated the effects of several carcinogens on the primary cilium of the RPTEC/TERT1 human proximal tubular epithelial cell line. Using both scanning electron microscopy and immunofluorescent labeling of the ciliary markers acetylated tubulin and Arl13b, we confirmed that RPTEC/TERT1 cells express primary cilium upon reaching confluence. Treatment with the carcinogens ochratoxin A (OTA) and potassium bromate (KBrO3) caused a significant reduction in the number of ciliated cells, while exposure to nifedipine, a noncarcinogenic renal toxin, had no effect on primary cilium expression. Flow cytometric analysis of the effects of all three compounds on the cell cycle revealed that only KBrO3 resulted in an increase in the proportion of cells entering the cell cycle. Microarray analysis revealed dysregulation of multiple pathways affecting ciliogenesis and ciliary maintenance following OTA and KBrO3 exposure, which were unaffected by nifedipine exposure. The primary cilium represents a unique physical checkpoint with relevance to carcinogenesis. We have shown that the renal carcinogens OTA and KBrO3 cause significant deciliation in a model of the proximal tubule. With KBrO3, this was followed by reentry into the cell cycle; however, deciliation was not found to be associated with reentry into the cell cycle following OTA exposure. Transcriptomic analysis identified dysregulation of Wnt signaling and ciliary trafficking in response to OTA and KBrO3 exposure. PMID:22262483

Biomaterials patterned with discontinuous microwalls for vascular smooth muscle cell culture: biodegradable small diameter vascular grafts and stable cell culture substrates.

PubMed

Heath, Daniel E; Kang, Gavin C W; Cao, Ye; Poon, Yin Fun; Chan, Vincent; Chan-Park, Mary B

2016-10-01

The medial layer of small diameter blood vessels contains circumferentially aligned vascular smooth muscle cells (vSMC) that possess contractile phenotype. In tissue-engineered constructs, these cellular characteristics are usually achieved by seeding planar scaffolds with vSMC, rolling the cell-laden scaffold into a tubular structure, and maturing the construct in a pulsatile bioreactor, a lengthy process that can take up to two months. During the maturation phase, the cells circumferentially orient, their contractile protein expression increases, and they obtain a contractile phenotype. Generating cell culture platforms that enable the rapid production of directionally oriented vSMC with increased contractile protein expression would be a major step forward for blood vessel tissue engineering and would greatly facilitate the in vitro study of vSMC biology. Previously, we developed a micropatterned cell culture surface that promotes orientation and contractile protein expression of vSMC. Herein, we explore two potential applications of this technology. First, we fabricate tubular and biodegradable scaffolds that possess the micropatterning on their exterior surface. When vSMC are seeded on these scaffolds, they initially proliferate in order to fill the microchannels and as confluence is reached the cells align in the direction of the micropatterning resulting in a biodegradable scaffold that is inhabited by circumferentially aligned vSMC within a week. Second, we illustrate that we can generate biostable cell culture surfaces that allow the in vitro study of the cells in a more contractile state. Specifically, we explore contractile protein expression of cells cultured on the micropatterned surfaces with the addition of soluble transforming growth factor beta one (TGFβ1).

1. WATER ENTERING CONFLUENCE POOL FROM BEAR CREEK AT LEFT, ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

1. WATER ENTERING CONFLUENCE POOL FROM BEAR CREEK AT LEFT, AND FROM SANTA ANA RIVER THROUGH TUNNEL #0 AT RIGHT. VIEW TO NORTHEAST. - Santa Ana River Hydroelectric System, Bear Creek Diversion Dam & Confluence Pool, Redlands, San Bernardino County, CA

Hyaluronan-Based Therapy for Metastatic Prostate Cancer

DTIC Science & Technology

2015-07-01

8217-AAG ACC CTT GTG CTC GTT GT-3’ and re- verse 5’-AGG TGG ACA CAA TCC CTC TG-3’ actin, forward 5’-TCC CTG GAG AAG AGC TAC GA-3’ and reverse 5’-AGC ACT... GTG TTG GCG TAC AG-3’. 2.2.5. Death-inducing signaling complex immunoprecipitation After HT-29 cells achieved 80% confluence, the cells were pretreated

Mechanisms of palatal epithelial seam disintegration by Transforming Growth Factor (TGF)-β3

PubMed Central

Ahmed, Shaheen; Liu, Chang-Chih; Nawshad, Ali

2007-01-01

TGFβ3 signaling initiates and completes sequential phases of cellular differentiation that is required for complete disintegration of the palatal medial edge seam, that progresses between 14 to 17 embryonic days in the murine system, which is necessary in establishing confluence of the palatal stroma. Understanding the cellular mechanism of palatal MES disintegration in response to TGFβ3 signaling will result in new approaches to defining the causes of cleft palate and other facial clefts that may result from failure of seam disintegration. We have isolated MES primary cells to study the details of MES disintegration mechanism by TGFβ3 during palate development using several biochemical and genetic approaches. Our results demonstrate a novel mechanism of MES disintegration where MES, independently yet sequentially, undergoes cell cycle arrest, cell migration and apoptosis to generate immaculate palatal confluency during palatogenesis in response to robust TGFβ3 signaling. The results contribute to a missing fundamental element to our base knowledge of the diverse roles of TGFβ3 in functional and morphological changes that MES undergo during palatal seam disintegration. We believe that our findings will lead to more effective treatment of facial clefting. PMID:17698055

75 FR 29238 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-25

... River). with Pot Ripple Creek Henry County. to approximately 0.5 mile upstream of the confluence with Pot Ripple Creek. Boiling Branch (Backwater effects From the confluence None +481 Unincorporated Areas... Drennon Road. Pot Ripple Creek (Backwater effects From the confluence None +488 Unincorporated Areas of...

75 FR 23600 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-04

... feet +571 upstream of the confluence with Millers Creek. Dale Hollow Lake (Wolf River) At the confluence with +663 Unincorporated Areas of the Wolf River. Clinton County. Approximately 1,800 feet +663 upstream of the confluence with Spring Creek. Lake Cumberland Just upstream of the Wolf +760 Unincorporated...

76 FR 35119 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2011-06-16

... (backwater effects from From the confluence with Pot +488 Unincorporated Areas of Kentucky River). Ripple Creek to Henry County. approximately 0.5 mile upstream of the confluence with Pot Ripple Creek. Boiling... Ripple Creek (backwater effects From the confluence with the +488 Unincorporated Areas of from Kentucky...

Production of bovine cloned embryos with donor cells frozen at a slow cooling rate in a conventional freezer (20 C)

USGS Publications Warehouse

Chacon, L.; Gomez, M.C.; Jenkins, J.A.; Leibo, S.P.; Wirtu, G.; Dresser, B.L.; Pope, C.E.

2009-01-01

Summary Usually, fibroblasts are frozen in dimethyl sulphoxide (DMSO, 10% v/v) at a cooling rate of 1 C/min in a low-temperature (80 C) freezer (LTF) before storage in liquid nitrogen (LN2); however, a LTF is not always available. The purpose of the present study was to evaluate apoptosis and viability of bovine fibroblasts frozen in a LTF or conventional freezer (CF; 20 C) and their subsequent ability for development to blastocyst stage after fusion with enucleated bovine oocytes. Percentages of live cells frozen in LTF (49.5%) and CF (50.6%) were similar, but significantly less than non-frozen control (88%). In both CF and LTF, percentages of live apoptotic cells exposed to LN2 after freezing were lower (4% and 5%, respectively) as compared with unexposed cells (10% and 18%, respectively). Cells frozen in a CF had fewer cell doublings/24 h (0.45) and required more days (9.1) to reach 100% confluence at the first passage (P) after thawing and plating as compared with cells frozen in a LTF (0.96 and 4.0 days, respectively). Hypoploidy at P12 was higher than at P4 in cells frozen in either a CF (37.5% vs. 19.2%) or in a LTF (30.0% vs. 15.4%). A second-generation cryo-solution reduced the incidence of necrosis (29.4%) at 0 h after thawing as compared with that of a first generation cryo-solution (DMEM + DMSO, 60.2%). The percentage of apoptosis in live cells was affected by cooling rate (CF = 1.9% vs. LFT = 0.7%). Development of bovine cloned embryos to the blastocyst stage was not affected by cooling rate or freezer type. ?? 2009 Cambridge University Press.

The effect of Platelet Lysate on osteoblast proliferation associated with a transient increase of the inflammatory response in bone regeneration.

PubMed

Ruggiu, Alessandra; Ulivi, Valentina; Sanguineti, Francesca; Cancedda, Ranieri; Descalzi, Fiorella

2013-12-01

Platelet Lysate (PL) contains a cocktail of growth factors and cytokines, which actively participates in tissue repair and its clinical application has been broadly described. The aim of this study was to assess the regenerative potential of PL for bone repair. We demonstrated that PL stimulation induces a transient increase of the inflammatory response in quiescent human osteoblasts, via NF-kB activation, COX-2 induction, PGE2 production and secretion of pro-inflammatory cytokines. Furthermore, we showed that long-term PL stimulation enhances proliferation of actively replicating osteoblasts, without affecting their differentiation potential, along with changes of cell morphology, resulting in increased cell density at confluence. In confluent resting osteoblasts, PL treatment induced resumption of proliferation, change in cell morphology and increase of cell density at confluence. A burst of PL treatment (24-h) was sufficient to trigger such processes in both conditions. These results correlated with up-regulation of the proliferative and survival pathways ERKs and Akt and with cell cycle re-activation via induction of CyclinD1 and phosphorylation of Rb, following PL stimulation. Our findings demonstrate that PL treatment results in activation and expansion of resting osteoblasts, without affecting their differentiation potential. Therefore PL represents a good therapeutic candidate in regenerative medicine for bone repair. Copyright © 2013 Elsevier Ltd. All rights reserved.

78 FR 21273 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2013-04-10

... Docket No.: FEMA-B-1117 Four Mile Run At the confluence with +10 Arlington County. the Potomac River... from From the confluence with +40 Arlington County. Potomac River). the Potomac River to a point located approximately 112 feet downstream of Chain Bridge Road. Potomac River At the confluence with +10...

76 FR 8978 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2011-02-16

... White River ted Areas effects from confluence of Taney White River). to County. approximate ly 685 feet upstream of the White River confluence Bee Creek From the None +698 Unincorpora (backwater White River ted Areas effects from confluence of Taney White River). to County. approximate ly 1,700 feet upstream of...

The Nature of Creativity: Cognitive and Confluence Perspectives

ERIC Educational Resources Information Center

Megalakaki, Olga; Craft, Anna; Cremin, Teresa

2012-01-01

In the present psychology-informed literature review we address some aspects of the nature of creativity from cognitive and confluence perspectives. The authors begin by discussing models of creativity offered by cognitive and confluence approaches, focusing on the transition from univariate to multivariate models. The article explores what these…

Lineage mapper: A versatile cell and particle tracker

NASA Astrophysics Data System (ADS)

Chalfoun, Joe; Majurski, Michael; Dima, Alden; Halter, Michael; Bhadriraju, Kiran; Brady, Mary

2016-11-01

The ability to accurately track cells and particles from images is critical to many biomedical problems. To address this, we developed Lineage Mapper, an open-source tracker for time-lapse images of biological cells, colonies, and particles. Lineage Mapper tracks objects independently of the segmentation method, detects mitosis in confluence, separates cell clumps mistakenly segmented as a single cell, provides accuracy and scalability even on terabyte-sized datasets, and creates division and/or fusion lineages. Lineage Mapper has been tested and validated on multiple biological and simulated problems. The software is available in ImageJ and Matlab at isg.nist.gov.

Flow structure at an ice-covered river confluence

NASA Astrophysics Data System (ADS)

Martel, Nancy; Biron, Pascale; Buffin-Bélanger, Thomas

2017-04-01

River confluences are known to exhibit complex relationships between flow structure, sediment transport and bed-form development. Flow structure at these sites is influenced by the junction angle, the momentum flux ratio (Mr) and bed morphology. In cold regions where an ice cover is present for most of the winter period, the flow structure is also likely affected by the roughness effect of the ice. However, very few studies have examined the impact of an ice cover on the flow structure at a confluence. The aims of this study are (1) to describe the evolution of an ice cover at a river confluence and (2) to characterize and compare the flow structure at a river confluence with and without an ice cover. The field site is a medium-sized confluence (around 40 m wide) between the Mit is and Neigette Rivers in the Bas-Saint-Laurent region, Quebec (Canada). The confluence was selected because a thick ice cover is present for most of the winter allowing for safe field work. Two winter field campaigns were conducted in 2015 and 2016 to obtain ice cover measurements in addition to hydraulic and morphological measurements. Daily monitoring of the evolution of the ice cover was made with a Reconyx camera. Velocity profiles were collected with an acoustic Doppler current profiler (ADCP) to reconstruct the three-dimensional flow structure. Time series of photographs allow the evolution of the ice cover to be mapped, linking the processes leading to the formation of the primary ice cover for each year. The time series suggests that these processes are closely related with both confluence flow zones and hydro-climatic conditions. Results on the thickness of the ice cover from in situ measurements reveal that the ice thickness tends to be thinner at the center of the confluence where high turbulent exchanges take place. Velocity measurements reveal that the ice cover affects velocity profiles by moving the highest velocities towards the center of the profiles. A spatio-temporal conceptual model is presented to illustrate the main differences on the three-dimensional flow structure at the river confluence with and without the ice cover.

Role of Stat3 and ErbB2 in Breast Cancer

DTIC Science & Technology

2012-10-01

also activated by receptor tyrosine kinases, such as the epidermal growth factor receptor (EGFR) or platelet -derived growth factor receptor (PDGFR...cells were grown to different densities, up to 5 days post-confluence, as indicated. Detergent cell lysates were probed for Stat3-ptyr705, active Rac...and lysates probed for total cav1, cadherin 11 or tubulin as a loading control. 15 C Figure 7: Cadherin 11 and Rac1 downregulation

Androgen receptor (AR) degradation enhancer ASC-J9® in an FDA-approved formulated solution suppresses castration resistant prostate cancer cell growth.

PubMed

Cheng, Max A; Chou, Fu-Ju; Wang, Keliang; Yang, Rachel; Ding, Jie; Zhang, Qiaoxia; Li, Gonghui; Yeh, Shuyuan; Xu, Defeng; Chang, Chawnshang

2018-03-28

ASC-J9 ® is a recently-developed androgen receptor (AR)-degradation enhancer that effectively suppresses castration resistant prostate cancer (PCa) cell proliferation and invasion. The optimal half maximum inhibitory concentrations (IC 50 ) of ASC-J9 ® at various PCa cell confluences (20%, 50%, and 100%) were assessed via both short-term MTT growth assays and long-term clonogenic proliferation assays. Our results indicate that the IC 50 values for ASC-J9 ® increased with increasing cell confluency. The IC 50 values were significantly decreased in PCa AR-positive cells compared to PCa AR-negative cells or in normal prostate cells. This suggests that ASC-J9 ® may function mainly via targeting the AR-positive PCa cells with limited unwanted side-effects to suppress the surrounding normal prostate cells. Mechanism dissection indicated that ASC-J9 ® might function via altering the apoptosis signals to suppress the PCa AR-negative PC-3 cells. Preclinical studies using multiple in vitro PCa cell lines and an in vivo mouse model with xenografted castration-resistant PCa CWR22Rv1 cells demonstrated that ASC-J9 ® has similar AR degradation effects when dissolved in FDA-approved solvents, including DMSO, PEG-400:Tween-80 (95:5), DMA:Labrasol:Tween-80 (10:45:45), and DMA:Labrasol:Tween-20 (10:45:45). Together, results from preclinical studies suggest a potential new therapy with AR-degradation enhancer ASC-J9 ® may potentially be ready to be used in human clinical trials in order to better suppress PCa at later castration resistant stages. Copyright © 2017 Elsevier B.V. All rights reserved.

Effects of cadmium on intercellular junctions in a renal epithelial cell line grown on permeable membrane supports

DOE Office of Scientific and Technical Information (OSTI.GOV)

Prozialeck, W.C.; Niewenhuis, R.J.

1991-03-11

Recent findings from the authors laboratories have shown that Cd{sup 2+} has relatively specific damaging effects on adhering and occluding junctions in the established porcine renal epithelial cell line, LLC-PK{sub 1}. The present studies were undertaken in order to further characterize the junction-perturbing effects of Cd{sup 2+} in polarized monolayers of LLC-PK{sub 1} cells, and to begin to identify the mechanisms underlying these effects. LLC-PK{sub 1} cells were grown to confluency on Millicell HA chambers and exposed to Cd{sup 2+} in polarized monolayers of LLC-PK{sub 1} cells, and to begin to identify the mechanisms underlying these effects. LLC-PK{sub 1} cellsmore » were grown to confluency on Millicell HA chambers an exposed to Cd{sup 2+} by adding CdCl{sub 2} to the solutions on either side of the cell monolayer. The integrity of cell-cell junctions was assessed by monitoring the transepithelial electrical resistance. The results showed that exposure to Cd{sup 2+} caused a pronounced decrease in transepithelial resistance without causing the cells to detach from the Millicell membrane. This decrease in resistance occurred more quickly and was much more pronounced when Cd{sup 2+} was added to the basolateral surface rather than the apical surface. Furthermore, the effects of Cd{sup 2+} were greatly reduced when excess Ca{sup 2+} was present in the medium. These results suggest that Cd{sup 2+} was present in the medium. These results suggest that Cd{sup 2+} may disrupt cell-cell junctions by interacting with Ca{sup 2+} binding sites or Ca{sup 2+} channels that are oriented toward the basolateral cell surface.« less

Arsenite induces apoptosis in human mesenchymal stem cells by altering Bcl-2 family proteins and by activating intrinsic pathway

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yadav, Santosh; Shi Yongli; Wang Feng

2010-05-01

Purpose: Environmental exposure to arsenic is an important public health issue. The effects of arsenic on different tissues and organs have been intensively studied. However, the effects of arsenic on bone marrow mesenchymal stem cells (MSCs) have not been reported. This study is designed to investigate the cell death process caused by arsenite and its related underlying mechanisms on MSCs. The rationale is that absorbed arsenic in the blood circulation can reach to the bone marrow and may affect the cell survival of MSCs. Methods: MSCs of passage 1 were purchased from Tulane University, grown till 70% confluency level andmore » plated according to the experimental requirements followed by treatment with arsenite at various concentrations and time points. Arsenite (iAs{sup III}) induced cytotoxic effects were confirmed by cell viability and cell cycle analysis. For the presence of canonic apoptosis markers; DNA damage, exposure of intramembrane phosphotidylserine, protein and m-RNA expression levels were analyzed. Results: iAs{sup III} induced growth inhibition, G2-M arrest and apoptotic cell death in MSCs, the apoptosis induced by iAs{sup III} in the cultured MSCs was, via altering Bcl-2 family proteins and by involving intrinsic pathway. Conclusion: iAs{sup III} can induce apoptosis in bone marrow-derived MSCs via Bcl-2 family proteins, regulating intrinsic apoptotic pathway. Due to the multipotency of MSC, acting as progenitor cells for a variety of connective tissues including bone, adipose, cartilage and muscle, these effects of arsenic may be important in assessing the health risk of the arsenic compounds and understanding the mechanisms of arsenic-induced harmful effects.« less

The Mississippi River: A place for fish

USGS Publications Warehouse

Schramm, Harold; Ickes, Brian; Chen, Yushun; Chapman, Duane C.; Jackson, John; Chen, Daqing; Li, Zhongjie; Kilgore, Jack; Phelps, Quinton E.; Eggleton, Michael

2016-01-01

The Mississippi River flows 3,734 km from its source at Lake Itasca, Minnesota to its outlet at the Gulf of Mexico. Along its course, it collects water from portions of two Canadian provinces and 41 % of the conterminous United States. Although greatly altered for navigation and flood control throughout much of its length, the Mississippi River remains an important fishery resource that provides habitat for 188 species of fishes and recreational and commercial fishing opportunities. The objectives of this chapter are to describe the contemporary fisheries habitat throughout the Mississippi River, identify how management to achieve human benefits influences the fishes and their habitats, and summarize efforts to conserve and enhance fish habitat. The 826-km headwater reach is entirely in Minnesota and remains largely unaltered. The reaches that extend 1,059 km from St. Anthony Falls, Minnesota to above the confluence with the Missouri River near St. Louis, Missouri have been altered by impoundment that has affected floodplain function, increased sedimentation of backwaters, and homogenized the formerly diverse aquatic habitats. After the confluence with the Missouri River, the Mississippi River flows freely for 1,849 km to the Gulf of Mexico. The alterations of the free-flowing reaches of greatest significance to the fisheries resource are reducing the duration and height of the flood pulse as a consequence of shortening the river channel, disconnection of the river from its historic and present floodplain, and loss of secondary channel-island complexes. Engineering features to improve commercial navigation have also added habitat and, when wisely manipulated, can be used to rehabilitate habitat. Some aspects of water quality have improved, but legacy chemicals and nutrient-laden inflows and sediments remain problems. Although true restoration in the sense of restoring all environmental conditions to an unaltered state is unlikely, the future value of the Mississippi River as a fisheries resource will depend on actively maintaining diverse and accessible aquatic habitats to support food webs and water quality suitable for fishes.

Summary of survival data from juvenile coho salmon in the Klamath River, northern California, 2009

USGS Publications Warehouse

Beeman, John W.; Juhnke, Steven D.

2009-01-01

A study of the effects of the discharge from Iron Gate Dam on the Klamath River on juvenile coho salmon during their seaward migration began in 2005. Estimates of fish survival through various reaches of the river downstream of the dam were completed in 2006, 2007, 2008, and 2009. This report describes the estimates of survival during 2009, and is a complement to similar reports for 2006, 2007, and 2008. For each year, a series of numerical models were evaluated to determine apparent survival and recapture probabilities of radio-tagged fish in several river reaches between Iron Gate Hatchery at river kilometer 309 and a site at river kilometer 33. The evaluations indicate that the primary differences among years are in the survivals through reaches upstream of the confluence of the Scott River with the Klamath River. Data from 2009, one of two years when fish from both hatchery and wild origins were available for analysis, indicate that survival of wild and hatchery fish are similar.

Thermal profiles for selected river reaches of the Methow and Chewuch Rivers, Washington, August 2011

USGS Publications Warehouse

Gendaszek, Andrew S.

2012-01-01

Longitudinal profiles of near-streambed and near-surface temperatures were collected for selected reaches of the Methow and Chewuch Rivers, Washington, during August 2011 to facilitate development of a stream temperature model near the confluence of the Methow and Chewuch Rivers. Temperature was measured using a probe with an internal datalogger towed behind a watercraft moving downstream at ambient river velocity. For the Methow River, an additional temperature survey was completed using near-streambed and near-surface probes towed behind a second watercraft that traversed the channel to measure vertical and lateral temperature variability. All data were referenced to location that was concurrently measured with a Global Positioning System. Data are presented as Microsoft Excel® files consisting of date and time, water temperature, and Washington State Plane North easting and northing.

Layering PLGA-based electrospun membranes and cell sheets for engineering cartilage-bone transition.

PubMed

Mouthuy, P-A; El-Sherbini, Y; Cui, Z; Ye, H

2016-04-01

It is now widely acknowledged that implants that have been designed with an effort towards reconstructing the transition between tissues might improve their functionality and integration in vivo. This paper contributes to the development of improved treatment for articular cartilage repair by exploring the potential of the combination of electrospinning technology and cell sheet engineering to create cartilage tissue. Poly(lactic-co-glycolic acid) (PLGA) was used to create the electrospun membranes. The focus being on the cartilage-bone transition, collagen type I and hydroxyapatite (HA) were also added to the scaffolds to increase the histological biocompatibility. Human mesenchymal stem cells (hMSCs) were cultured in thermoresponsive dishes to allow non-enzymatic removal of an intact cell layer after reaching confluence. The tissue constructs were created by layering electrospun membranes with sheets of hMSCs and were cultured under chondrogenic conditions for up to 21 days. High viability was found to be maintained in the multilayered construct. Under chondrogenic conditions, reverse-transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry have shown high expression levels of collagen type X, a form of collagen typically found in the calcified zone of articular cartilage, suggesting an induction of chondrocyte hypertrophy in the PLGA-based scaffolds. To conclude, this paper suggests that layering electrospun scaffolds and cell sheets is an efficient approach for the engineering of tissue transitions, and in particular the cartilage-bone transition. The use of PLGA-based scaffold might be particularly useful for the bone-cartilage reconstruction, since the differentiated tissue constructs seem to show characteristics of calcified cartilage. Copyright © 2013 John Wiley & Sons, Ltd.

Apoptotic effect of α-Fe2O3 and SiO2 nanoparticles in human rhabdomyosarcoma cell line

NASA Astrophysics Data System (ADS)

Fatima, Mahvish; Fakhar-e-Alam, Muhammad; Atif, M.; Nadeem Shakoor, Muhammad; Afzal, Muhammad; Waseem, Muhammad; Hammad Aziz, Muhammad

2014-12-01

Nanotechnology provides the opportunity for the development of new materials in the nanometer size range, with many potential applications in biological sciences and clinical medicine. It has been reported that RD (muscle cancer cell line) is the most common soft tissue sarcoma in children originating from immature cells, comprising 2.9% of all malignancies in patients younger than 20 years old, with 350 cases diagnosed annually in the United States. Soft tissue is the most common target organ for nanoparticles after they gain significant entry into the target site through any of the possible routes. RD cell lines have been used as an experimental biological model in this article. A suitable environment was provided until 75% of RD cell confluence was reached. Prior to determination of toxicity of hematite (α-Fe2O3) and SiO2 nanoparticles, the sizes and shapes were confirmed using scanning electron microscopy (SEM), and the sizes were about 66 and 250 nm respectively. Moreover, 10-80 μg ml-1 of α-Fe2O3 and SiO2 nanoparticles dispersed in solution were labeled for each row of 96 well plates. The present study evaluates the suppression factor of the said particles, which leads to cell killing phenomena. After successful measurements in the above mentioned experiment, the author will be able to give the actual cause of cell killing effects. The given study has provided valuable insights into a feasible mechanism of apoptosis caused by α-Fe2O3 and SiO2 nanoparticles. An underlying promising mechanism of apoptosis due to α-Fe2O3 and SiO2 nanoparticle exposure should be further investigated at the in vivo level.

The confluence model: birth order as a within-family or between-family dynamic?

PubMed

Zajonc, R B; Sulloway, Frank J

2007-09-01

The confluence model explains birth-order differences in intellectual performance by quantifying the changing dynamics within the family. Wichman, Rodgers, and MacCallum (2006) claimed that these differences are a between-family phenomenon--and hence are not directly related to birth order itself. The study design and analyses presented by Wichman et al. nevertheless suffer from crucial shortcomings, including their use of unfocused tests, which cause statistically significant trends to be overlooked. In addition, Wichman et al. treated birth-order effects as a linear phenomenon thereby ignoring the confluence model's prediction that these two samples may manifest opposing results based on age. This article cites between- and within-family data that demonstrate systematic birth-order effects as predicted by the confluence model. The corpus of evidence invoked here offers strong support for the assumption of the confluence model that birth-order differences in intellectual performance are primarily a within-family phenomenon.

Bovine dedifferentiated adipose tissue (DFAT) cells

PubMed Central

Wei, Shengjuan; Du, Min; Jiang, Zhihua; Duarte, Marcio S; Fernyhough-Culver, Melinda; Albrecht, Elke; Will, Katja; Zan, Linsen; Hausman, Gary J; Elabd, Elham M Youssef; Bergen, Werner G; Basu, Urmila; Dodson, Michael V

2013-01-01

Dedifferentiated fat cells (DFAT cells) are derived from lipid-containing (mature) adipocytes, which possess the ability to symmetrically or asymmetrically proliferate, replicate, and redifferentiate/transdifferentiate. Robust cell isolation and downstream culture methods are needed to isolate large numbers of DFAT cells from any (one) adipose depot in order to establish population dynamics and regulation of the cells within and across laboratories. In order to establish more consistent/repeatable methodology here we report on two different methods to establish viable DFAT cell cultures: both traditional cell culture flasks and non-traditional (flat) cell culture plates were used for ceiling culture establishment. Adipocytes (maternal cells of the DFAT cells) were easier to remove from flat culture plates than flasks and the flat plates also allowed cloning rings to be utilized for cell/cell population isolation. While additional aspects of usage of flat-bottomed cell culture plates may yet need to be optimized by definition of optimum bio-coating to enhance cell attachment, utilization of flat plate approaches will allow more efficient study of the dedifferentiation process or the DFAT progeny cells. To extend our preliminary observations, dedifferentiation of Wagyu intramuscular fat (IMF)-derived mature adipocytes and redifferentiation ability of DFAT cells utilizing the aforementioned isolation protocols were examined in traditional basal media/differentiation induction media (DMI) containing adipogenic inducement reagents. In the absence of treatment approximately 10% isolated Wagyu IMF-mature adipocytes dedifferentiated spontaneously and 70% DFAT cells displayed protracted adipogenesis 12 d after confluence in vitro. Lipid-free intracellular vesicles in the cytoplasm (vesicles possessing an intact membrane but with no any observable or stainable lipid inside) were observed during redifferentiation. One to 30% DFAT cells redifferentiated into lipid-assimilating adipocytes in the DMI media, with distinct lipid-droplets in the cytoplasm and with no observable lipid-free vesicles inside. Moreover, a high confluence level promoted the redifferentiation efficiency of DFAT cells. Wagyu IMF dedifferentiated DFAT cells exhibited unique adipogenesis modes in vitro, revealing a useful cell model for studying adipogenesis and lipid metabolism. PMID:23991361

Automated method for the rapid and precise estimation of adherent cell culture characteristics from phase contrast microscopy images.

PubMed

Jaccard, Nicolas; Griffin, Lewis D; Keser, Ana; Macown, Rhys J; Super, Alexandre; Veraitch, Farlan S; Szita, Nicolas

2014-03-01

The quantitative determination of key adherent cell culture characteristics such as confluency, morphology, and cell density is necessary for the evaluation of experimental outcomes and to provide a suitable basis for the establishment of robust cell culture protocols. Automated processing of images acquired using phase contrast microscopy (PCM), an imaging modality widely used for the visual inspection of adherent cell cultures, could enable the non-invasive determination of these characteristics. We present an image-processing approach that accurately detects cellular objects in PCM images through a combination of local contrast thresholding and post hoc correction of halo artifacts. The method was thoroughly validated using a variety of cell lines, microscope models and imaging conditions, demonstrating consistently high segmentation performance in all cases and very short processing times (<1 s per 1,208 × 960 pixels image). Based on the high segmentation performance, it was possible to precisely determine culture confluency, cell density, and the morphology of cellular objects, demonstrating the wide applicability of our algorithm for typical microscopy image processing pipelines. Furthermore, PCM image segmentation was used to facilitate the interpretation and analysis of fluorescence microscopy data, enabling the determination of temporal and spatial expression patterns of a fluorescent reporter. We created a software toolbox (PHANTAST) that bundles all the algorithms and provides an easy to use graphical user interface. Source-code for MATLAB and ImageJ is freely available under a permissive open-source license. © 2013 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

Automated Method for the Rapid and Precise Estimation of Adherent Cell Culture Characteristics from Phase Contrast Microscopy Images

PubMed Central

Jaccard, Nicolas; Griffin, Lewis D; Keser, Ana; Macown, Rhys J; Super, Alexandre; Veraitch, Farlan S; Szita, Nicolas

2014-01-01

The quantitative determination of key adherent cell culture characteristics such as confluency, morphology, and cell density is necessary for the evaluation of experimental outcomes and to provide a suitable basis for the establishment of robust cell culture protocols. Automated processing of images acquired using phase contrast microscopy (PCM), an imaging modality widely used for the visual inspection of adherent cell cultures, could enable the non-invasive determination of these characteristics. We present an image-processing approach that accurately detects cellular objects in PCM images through a combination of local contrast thresholding and post hoc correction of halo artifacts. The method was thoroughly validated using a variety of cell lines, microscope models and imaging conditions, demonstrating consistently high segmentation performance in all cases and very short processing times (<1 s per 1,208 × 960 pixels image). Based on the high segmentation performance, it was possible to precisely determine culture confluency, cell density, and the morphology of cellular objects, demonstrating the wide applicability of our algorithm for typical microscopy image processing pipelines. Furthermore, PCM image segmentation was used to facilitate the interpretation and analysis of fluorescence microscopy data, enabling the determination of temporal and spatial expression patterns of a fluorescent reporter. We created a software toolbox (PHANTAST) that bundles all the algorithms and provides an easy to use graphical user interface. Source-code for MATLAB and ImageJ is freely available under a permissive open-source license. Biotechnol. Bioeng. 2014;111: 504–517. © 2013 Wiley Periodicals, Inc. PMID:24037521

Inhibition of histone deacetylation and DNA methylation improves gene expression mediated by the adeno-associated virus/phage in cancer cells.

PubMed

Kia, Azadeh; Yata, Teerapong; Hajji, Nabil; Hajitou, Amin

2013-10-22

Bacteriophage (phage), viruses that infect bacteria only, have become promising vectors for targeted systemic delivery of genes to cancer, although, with poor efficiency. We previously designed an improved phage vector by incorporating cis genetic elements of adeno-associated virus (AAV). This novel AAV/phage hybrid (AAVP) specifically targeted systemic delivery of therapeutic genes into tumors. To advance the AAVP vector, we recently introduced the stress-inducible Grp78 tumor specific promoter and found that this dual tumor-targeted AAVP provides persistent gene expression, over time, in cancer cells compared to silenced gene expression from the CMV promoter in the parental AAVP. Herein, we investigated the effect of histone deacetylation and DNA methylation on AAVP-mediated gene expression in cancer cells and explored the effect of cell confluence state on AAVP gene expression efficacy. Using a combination of AAVP expressing the GFP reporter gene, flow cytometry, inhibitors of histone deacetylation, and DNA methylation, we have demonstrated that histone deacetylation and DNA methylation are associated with silencing of gene expression from the CMV promoter in the parental AAVP. Importantly, inhibitors of histone deacetylases boost gene expression in cancer cells from the Grp78 promoter in the dual tumor-targeted AAVP. However, cell confluence had no effect on AAVP-guided gene expression. Our findings prove that combination of histone deacetylase inhibitor drugs with the Grp78 promoter is an effective approach to improve AAVP-mediated gene expression in cancer cells and should be considered for AAVP-based clinical cancer gene therapy.

Novel Function of NIBP in Breast Cancer

DTIC Science & Technology

2012-05-01

reduced in NIBP knockdown cells (Fig. 8). 7 pRK -Flag-NIBP Isoforms(aa) 960 944 1200 1246 1148 S E A P A c ti v it y ( F o ld...Fig.9. MDA-MB-231 cells were co-transfected by TurboFectin8.0 with empty pRK -Flag vector or various isoforms of NIBP with NF-B-SEAP reporter and...Ser536) pRK -Flag 12060301550 NIBP-mutA 12060301550TNFα (min) Fig.11. MDA-MB-231 cells at 60% confluence in 6-well plates were transfected with empty

Evaluation of Confluence Model Variables on IQ and Achievement Test Scores in a Sample of 6- to 11-Year-Old Children.

ERIC Educational Resources Information Center

Svanum, Soren; Bringle, Robert G.

1980-01-01

The confluence model of cognitive development was tested on 7,060 children. Family size, sibling order within family sizes, and hypothesized age-dependent effects were tested. Findings indicated an inverse relationship between family size and the cognitive measures; age-dependent effects and other confluence variables were found to be…

An Experimental Study to Control Scour at River Confluence

NASA Astrophysics Data System (ADS)

Wuppukondur, A.; Chandra, V.

2015-12-01

The aim of present study is finding a method to control sediment erosion at river confluence. The confluences are mixture of two different flows and are common occurrences along the river. River confluences are sites of natural scour phenomenon and also influence reservoir sedimentation. The river confluence is associated with a separation zone, stagnation zone and a mixing layer along which the scour hole is observed. The eroded sediment creates potential problems by depositing at unwanted downstream locations such as barrages, weirs, check dams, reservoirs etc. As per the literature, the storage capacity of major reservoirs in India is going to be reduced nearly half of the storage capacity by 2020. Hence, an experimental study has been conducted on mobile bed (d50=0.28 mm) with a confluence angle of 90o for a discharge ratio (Qr) of 0.5, where, Qr is defined as the ratio between lateral flow discharge (Ql) and main flow discharge (Qm). Circular shape pile models of same diameter are arranged in a systematic manner with constant spacing (5 cm, 10 cm and 15 cm) to change the flow pattern for reducing scour at the confluence. Two types of pile models (8 mm ϕ and 12 mm ϕ) are used to conduct the experiments. The experimental results show that maximum scour depth at confluence is reduced by 60%. In addition, the bed profile modifications are also reported. Keywords: Reservoir sedimentation, River confluence, Mobile bed, Scour, Vanes. References:1. Borghei, S. M., and Sahebari, A. J. (2010). "Local Scour at Open-Channel Junctions", Journal of Hydraulic Research, 48(4), 37 - 41. 2. Kothyari, U. C. (1996). "Methods for Estimation Sediment Yield from Catchments", Proc., Int. Sem. On Civil Engg. Practices in Twenty First Century, Roorkee, India, 1071-1086. 3. Mosley, M. P. (1976) "An Experimental Study of Channel Confluences". The Journal of Geology, 84(55), 532-562. 4. Ouyang, H. T. (2009). "Investigation on the dimensions and shape of a submerged vane for sediment management in alluvial channels." Journal of Hydraulic Engineering, 135 (3), 209- 217. 5. Tan, S. K., Yu, G., Lim, S.Y., and Ong, M. C. (2005). "Flow structure and sediment motion around submerged vanes in open channel." Journal of Waterway, Port, Coastal and Ocean Engineering, 131(3), 132-136.

Characterizing Three-Dimensional Mixing Process in a River Confluence using Hydro-acoustical Backscatter and Flow Measurement

NASA Astrophysics Data System (ADS)

Son, Geunsoo; Kim, Dongsu; Kim, YoungDo; Lyu, Siwan; Kim, Seojun

2017-04-01

River confluences are zones where two rivers with different geomorphic and hydraulic characteristics amalgamate, resulting in rapid change in terms of flow regime, sediment entrainment and hydraulic geometry. In these confluence zones, the flow structure is basically complicated responded with concurrent mixing of physical and chemical aquatic properties, and continuous channel morphology could be changed due to erosion and sedimentation. In addition, the confluences are regions in which two rivers join and play an important role in river ecology. In order to characterize the mixing process of confluence for understanding the impacts of a river on the other river, therefore, it has been crucial to analyze the spatial mixing patterns for main streams depending on various inflow conditions of tributaries. However, most conventional studies have mostly relied upon hydraulic or water quality numerical models for understanding mixing pattern analysis of confluences, due to the difficulties to acquire a wide spatial range of in-situ data especially for characterizing this kind of mixing process. Even with intensive in-situ measurements, those researches tended to focus mainly on the hydraulic characteristics such as the flow and morphological complexity of confluence, so that very few studies comprehensively included sediment variation with flow at the same time. In this study, subsequently, flow and sediment mixing characteristics were concurrently investigated in the confluence between Nakdong and Nam river in South Korea, where it has been frequently questioned to determine how Nam river affects Nakdong river that recently have suffered various environmental problems such as green algae bloom and erosion/deposition in the confluence. We basically examined the mixing characteristics of confluence by using acoustic Doppler current profilers (ADCPs) which were used to measure hydraulic factors such as flow rate and depth, as well as measuring the suspended sediment concentration by using acoustic backscatter. Cross-sectional ADCP measurements in a confluence were collected with high spatial resolution in order to analyze the details of spatial distribution in the perspective of the three-dimensional mixing patterns of flow and sediment, where backscatters (or SNR) measured from ADCPs were used to track sediment mixing assuming that it could be a surrogate to estimate the suspended sediment concentration. Raw backscatter data were corrected by considering the beam spreading and absorption by water. Also, an optical Laser diffraction instrument (LISST) was used to verify the method of acoustic backscatter and to collect the particle size distribution of main stream and tributary. In addition, image-based spatial distributions of sediment mixture in the confluence were monitored in various flow conditions by using an unmanned aerial vehicle (UAV), which were compared with the spatial distribution of acoustic backscatter. As results, we found that when acoustic backscatter and flow measurements by ADCPs were well processed, they could be proper indicators to identify the spatial patterns of the three-dimensional mixing process between two rivers.

Differentiation of human umbilical cord mesenchymal stem cells into dermal fibroblasts in vitro

DOE Office of Scientific and Technical Information (OSTI.GOV)

Han, Yanfu; Chai, Jiake, E-mail: cjk304@126.com; Sun, Tianjun

2011-10-07

Highlights: {yields} Mesenchymal stem cells (MSCs) are potential seed cells for tissue-engineered skin. {yields} Tissue-derived umbilical cord MSCs (UCMSCs) can readily be isolated in vitro. {yields} We induce UCMSCs to differentiate into dermal fibroblasts via conditioned medium. {yields} Collagen type I and collagen type III mRNA level was higher in differentiated cells. {yields} UCMSCs-derived fibroblast-like cells strongly express fibroblast-specific protein. -- Abstract: Tissue-derived umbilical cord mesenchymal stem cells (UCMSCs) can be readily obtained, avoid ethical or moral constraints, and show excellent pluripotency and proliferation potential. UCMSCs are considered to be a promising source of stem cells in regenerative medicine. Inmore » this study, we collected newborn umbilical cord tissue under sterile conditions and isolated UCMSCs through a tissue attachment method. UCMSC cell surface markers were examined using flow cytometry. On the third passage, UCMSCs were induced to differentiate into dermal fibroblasts in conditioned induction media. The induction results were detected using immunofluorescence with a fibroblast-specific monoclonal antibody and real time PCR for type I and type III collagen. UCMSCs exhibited a fibroblast-like morphology and reached 90% confluency 14 to 18 days after primary culture. Cultured UCMSCs showed strong positive staining for CD73, CD29, CD44, CD105, and HLA-I, but not CD34, CD45, CD31, or HLA-DR. After differentiation, immunostaining for collagen type I, type III, fibroblast-specific protein, vimentin, and desmin were all strongly positive in induced cells, and staining was weak or negative in non-induced cells; total transcript production of collagen type I and collagen type III mRNA was higher in induced cells than in non-induced cells. These results demonstrate that UCMSCs can be induced to differentiate into fibroblasts with conditioned induction media and, in turn, could be used as seed cells for tissue-engineered dermis.« less

Developing Depositional Models for Mercury Contaminated Floodplain Deposits Using Geomorphic Mapping and GIS in South River, Virginia

NASA Astrophysics Data System (ADS)

Barbieri, A.; Pizzuto, J.; O'Neal, M. A.; Rhoades, E.

2007-12-01

Mercury was introduced into the South River from the 1930s to the 1950s from an industrial plant in Waynesboro, Virginia. Mercury contamination in fish tissue continues to exceed acceptable levels. The contaminated sediments in the river's floodplains are probably the present source of mercury to the South River ecosystem. Locating and determining the extent and depositional history of these deposits are important for understanding the mercury cycle in the river as well as for remediation plans. The South River is a sinuous, single thread alluvial river with frequent bedrock exposures along its bed and banks. Overbank deposits are discontinuous and thin. Rates of lateral migration by the South River are extremely low, averaging 0.02 m/yr, and the river has been influenced by mill dams along a 19 km study reach. This 19 km section of the 37 km river reach was selected for the study because of its high concentration of Hg. Six different categories of floodplain deposits dating from 1937-2005 have been identified throughout the river using studies of historical aerial photographs in a GIS framework, field mapping, dendro- and radionuclide dating, grain size and Hg analysis. Not surprisingly, traditional depositional models of meandering rivers do not apply. Floodplain depositional units include mill dam deposits, point bar/bench deposits, concave bank bench deposits, islands, cattle deposits, and tributary confluences deposits. The most important deposits for sequestering historic mercury are those that also store the most silt and clay. These include mill dam deposits, point bar/bench deposits, concave bank deposits, and tributary confluence deposits. Many of these deposits represent reservoirs of mercury-contaminated sediments that could supply significant amounts of mercury into the river presently and in the future.

Serum of patients with oral pemphigus vulgaris impairs keratinocyte wound repair in vitro: a time-lapse study on the efficacy of methylprednisolone and pyridostigmine bromide.

PubMed

Lanza, A; Stellavato, A; Heulfe, I; Landi, C; Gombos, F; Cirillo, N

2009-10-01

Pemphigus vulgaris (PV) is an autoimmune blistering disease affecting primarily oral mucosa and skin. Among the drugs used for the therapy of pemphigus, both methylprednisolone (MP) and pyridostigmine bromide (PBr) can prevent acantholysis in vitro. However, their putative therapeutic properties in regenerating PV-like lesions and promoting the healing process still remain to be demonstrated. To address this issue, here we have developed a model for studying the process of epithelial cleft regeneration in PV by artificially wounding keratinocyte monolayers. The experimental model was established by scratching confluent monolayers to simulate the epithelial cleft; then, wound regeneration in the presence of submaximal concentrations of PV sera was studied by time-lapse microscopy, with or without the addition of MP and PBr in the culture medium. Pemphigus vulgaris serum inhibited epithelial cleft repair of wounded monolayers. Indeed, in the presence of 10% (v/v) PV serum, keratinocytes reached only 2% confluence within 72 h vs an almost complete healing of controls. When administered together with PV sera, MP significantly (P < 0.01) enhanced wound fill by 30% after 72 h. PV-associated wound repair was significantly (P < 0.05) ameliorated by PBr by 24 h and keratinocytes reached 20% confluence after 72 h. Interestingly, neither MP nor PBr could accelerate wound healing when compared with untreated control monolayers. In PV, MP and PBr exert their curative effects in part by enhancing the regeneration properties of keratinocytes. Indeed, our data suggest that both drugs can specifically counterbalance the detrimental effects of PV serum on keratinocyte wound healing. These findings provide an explanation for the efficacy of MP and PBr in the treatment of PV lesions in human skin and oral mucosa.

A one-dimensional, steady-state, dissolved-oxygen model and waste-load assimilation study for Cedar Creek, Dekalb and Allen counties, Indiana

USGS Publications Warehouse

Wilber, William G.; Peters, J.G.; Ayers, M.A.; Crawford, Charles G.

1979-01-01

A digital model calibrated to conditions in Cedar Creek was used to develop alternatives for future waste loadings that would be compatible with Indiana stream water-quality standards defined for two critical hydrologic conditions, summer and winter low flows. The model indicates that the dissolved-oxygen concentration of the Auburn wastewater effluent and nitrification are the most significant factors affecting the dissolved-oxygen concentration in Cedar Creek during summer low flows. The observed dissolved-oxygen concentration of the Auburn wastewater effluent was low and averaged 30 percent of saturation. Projected nitrogenous biochemical-oxygen demand loads, from the Indiana State Board of Health, for the Auburn and Waterloo wastewater-treatment facilities will result in violations of the current instream dissolved-oxygen standard (5 mg/l), even with an effluent dissolved-oxygen concentration of 80 percent saturation. Natural streamflow for Cedar Creek upstream from the confluence of Willow and Little Cedar Creeks is small compared with the waste discharge, so benefits of dilution for Waterloo and Auburn are minimal. The model also indicates that, during winter low flows, ammonia toxicity, rather than dissolved oxygen, is the limiting water-quality criterion in the reach of Cedar Creek downstream from the wastewater-treatment facility at Auburn and the confluence of Garrett ditch. Ammonia-nitrogen concentrations predicted for 1978 through 2000 downstream from the Waterloo wastewater-treatment facility do not exceed Indiana water-quality standards for streams. Calculations of the stream 's assimilative capacity indicate that future waste discharge in the Cedar Creek basin will be limited to the reaches between the Auburn wastewater-treatment facility and County Road 68. (Kosco-USGS)

Glutathione Enhances Antibiotic Efficiency and Effectiveness of DNase I in Disrupting Pseudomonas aeruginosa Biofilms While Also Inhibiting Pyocyanin Activity, Thus Facilitating Restoration of Cell Enzymatic Activity, Confluence and Viability

PubMed Central

Das, Theerthankar; Simone, Martin; Ibugo, Amaye I.; Witting, Paul K.; Manefield, Mike; Manos, Jim

2017-01-01

Pyocyanin secreted by Pseudomonas aeruginosa is a virulence factor that damages epithelial cells during infection through the action of reactive oxygen species, however, little is known about its direct effect on biofilms. We demonstrated that pyocyanin-producing P. aeruginosa strains (PA14WT, DKN370, AES-1R, and AES-2) formed robust biofilms in contrast to the poorly formed biofilms of the pyocyanin mutant PA14ΔphzA-G and the low pyocyanin producer AES-1M. Addition of DNase I and reduced glutathione (GSH) significantly reduced biofilm biomass of pyocyanin-producing strains (P < 0.05) compared to non-pyocyanin producers. Subsequently we showed that a combined treatment comprising: GSH + DNase I + antibiotic, disrupted and reduced biofilm biomass up to 90% in cystic fibrosis isolates AES-1R, AES-2, LESB58, and LES431 and promoted lung epithelial cell (A549) recovery and growth. We also showed that exogenously added GSH restored A549 epithelial cell glutathione reductase activity in the presence of pyocyanin through recycling of GSSG to GSH and consequently increased total intracellular GSH levels, inhibiting oxidative stress, and facilitating cell growth and confluence. These outcomes indicate that GSH has multiple roles in facilitating a return to normal epithelial cell growth after insult by pyocyanin. With increased antibiotic resistance in many bacterial species, there is an urgency to establish novel antimicrobial agents. GSH is able to rapidly and comprehensively destroy P. aeruginosa associated biofilms while at a same time assisting in the recovery of host cells and re-growth of damaged tissue. PMID:29312161

Flow structure at low momentum ratio river confluences

NASA Astrophysics Data System (ADS)

Moradi, Gelare; Rennie, Colin. D.; Cardot, Romain; Mettra, François; Lane, Stuart. N.

2017-04-01

The flow structure at river confluences is a complex pattern of fluid motion and can be characterized by the formation of secondary circulation. As river confluences play an essential role on flow hydrodynamics and control the movement of sediment through river networks, there has been substantial attention given to this subject in recent decades. However, there is still much debate over how momentum ratio and sediment transport can control secondary circulation and mixing processes. In particular, studies have tended to assume that there is some equilibrium between the bed morphology present and the flow structures that form in the junction region. However, this overlooks the fact that tributaries may be associated with highly varying sediment supply regimes, especially for shorter and steeper tributaries, with temporal changes in sediment delivery ratios (between the main stem and the tributary) that do not follow exactly changes in momentum ratio. This may lead to bed morphologies that are a function of rates of historical sediment supply during sediment transporting events and not the momentum ratio associated with the junction during its measurement. It is quite possible that tributaries with low flow momentum ratio have a relatively higher sediment delivery ratio, such that the tributary is still able to influence significantly secondary circulation in the main channel, long after the sediment transport event, and despite its low flow momentum during measurement. The focus of this paper is low momentum ratio junctions where it is possible that the tributary can deliver large amounts of sediment. Secondary circulation at junctions is thought to be dominated by streamwise-oriented vortical cells. These cells are produced by the convergence of surface flow towards the centre of the main channel, with descending motion in the zone of maximum flow convergence. Once flow arrives at the bed, it diverges and completes its rotation by an upwelling motion through the surface at the channels margins. Numerical models, laboratory experiments and field studies have confirmed the presence of this motion. However, such studies have focused on situations where the momentum ratio is close to one and there have been fewer investigations of confluences where the momentum ratio is much less than one. This study presents field investigations in two upper Rhône river confluences in Switzerland, using an acoustic Doppler current profiler (aDcp). These two confluences are characterized by low momentum ratios but potentially higher sediment delivery ratios during extreme events. Results show that sediment delivery from the tributary during extreme events leads to the formation of a tributary mouth bar and associated bed discordance as well as a bank attached bar downstream of the tributary. In both cases, this discordant bed forms a two-layer flow and the water from the tributary penetrates into the upper part of the main river water column. This results in a mixing interface that is shifted toward the outer bank. When this mixing layer detaches from the tributary outer bank, it forms a large recirculation region in the upper part of the water column and a pronounced scour hole at this bank. The bank attached bar that forms downstream during sediment supply events leads to substantial curvature of the main channel flow, even when the flow momentum of the tributary is low and helps to shift the zone of deepest main river flow towards the outer bank.

Flood-inundation Maps for the Deerfield River, Franklin County, Massachusetts, from the Confluence with the Cold River Tributary to the Connecticut River

USGS Publications Warehouse

Lombard, Pamela J.; Bent, Gardner C.

2015-09-02

The availability of the flood-inundation maps at http://water.usgs.gov/osw/flood_inundation/, combined with information regarding current (near real-time) stage from the two U.S. Geological Survey streamgages in the study reach, can provide emergency management personnel and residents with information to aid in flood response activities, such as evacuations and road closures, and with postflood recovery efforts. The flood-inundation maps are nonregulatory, but provide Federal, State, and local agencies and the public with estimates of the potential extent of flooding during selected peak-flow events.

Convergence of advances in genomics, team science, and repositories as drivers of progress in psychiatric genomics.

PubMed

Lehner, Thomas; Senthil, Geetha; Addington, Anjené M

2015-01-01

After many years of unfilled promise, psychiatric genetics has seen an unprecedented number of successes in recent years. We hypothesize that the field has reached an inflection point through a confluence of four key developments: advances in genomics; the orientation of the scientific community around large collaborative team science projects; the development of sample and data repositories; and a policy framework for sharing and accessing these resources. We discuss these domains and their effect on scientific progress and provide a perspective on why we think this is only the beginning of a new era in scientific discovery. Published by Elsevier Inc.

Rethinking turbidite paleoseismology along the Cascadia subduction zone

USGS Publications Warehouse

Atwater, Brian F.; Carson, Bobb; Griggs, Gary B.; Johnson, H. Paul; Salmi, Marie

2014-01-01

A stratigraphic synthesis of dozens of deep-sea cores, most of them overlooked in recent decades, provides new insights into deep-sea turbidites as guides to earthquake and tsunami hazards along the Cascadia subduction zone, which extends 1100 km along the Pacific coast of North America. The synthesis shows greater variability in Holocene stratigraphy and facies off the Washington coast than was recognized a quarter century ago in a confluence test for seismic triggering of sediment gravity flows. That test compared counts of Holocene turbidites upstream and downstream of a deep-sea channel junction. Similarity in the turbidite counts among seven core sites provided evidence that turbidity currents from different submarine canyons usually reached the junction around the same time, as expected of widespread seismic triggering. The fuller synthesis, however, shows distinct differences between tributaries, and these differences suggest sediment routing for which the confluence test was not designed. The synthesis also bears on recent estimates of Cascadia earthquake magnitudes and recurrence intervals. The magnitude estimates hinge on stratigraphic correlations that discount variability in turbidite facies. The recurrence estimates require turbidites to represent megathrust earthquakes more dependably than they do along a flow path where turbidite frequency appears limited less by seismic shaking than by sediment supply. These concerns underscore the complexity of extracting earthquake history from deep-sea turbidites at Cascadia.

Chloride control and monitoring program in the Wichita River Basin, Texas, 1996-2009

USGS Publications Warehouse

Haynie, M.M.; Burke, G.F.; Baldys, Stanley

2011-01-01

Water resources of the Wichita River Basin in north-central Texas are vital to the water users in Wichita Falls, Tex., and surrounding areas. The Wichita River Basin includes three major forks of the Wichita River upstream from Lake Kemp, approximately 50 miles southwest of Wichita Falls, Tex. The main stem of the Wichita River is formed by the confluence of the North Wichita River and Middle Fork Wichita River upstream from Truscott Brine Lake. The confluence of the South Wichita River with the Wichita River is northwest of Seymour, Tex. (fig. 1). Waters from the Wichita River Basin, which is part of the Red River Basin, are characterized by high concentrations of chloride and other salinity-related constituents from salt springs and seeps (hereinafter salt springs) in the upper reaches of the basin. These salt springs have their origins in the Permian Period when the Texas Panhandle and western Oklahoma areas were covered by a broad shallow sea. Over geologic time, evaporation of the shallow seas resulted in the formation of salt deposits, which today are part of the geologic formations underlying the area. Groundwater in these formations is characterized by high chloride concentrations from these salt deposits, and some of this groundwater is discharged by the salt springs into the Wichita River.

Estuarine microbial food web patterns in a Lake Erie coastal wetland.

PubMed

Lavrentyev, P J; McCarthy, M J; Klarer, D M; Jochem, F; Gardner, W S

2004-11-01

Composition and distribution of planktonic protists were examined relative to microbial food web dynamics (growth, grazing, and nitrogen cycling rates) at the Old Woman Creek (OWC) National Estuarine Research Reserve during an episodic storm event in July 2003. More than 150 protistan taxa were identified based on morphology. Species richness and microbial biomass measured via microscopy and flow cytometry increased along a stream-lake (Lake Erie) transect and peaked at the confluence. Water column ammonium (NH4+) uptake (0.06 to 1.82 microM N h(-1)) and regeneration (0.04 to 0.55 microM N h(-1)) rates, measured using 15NH4+ isotope dilution, followed the same pattern. Large light/dark NH4+ uptake differences were observed in the hypereutrophic OWC interior, but not at the phosphorus-limited Lake Erie site, reflecting the microbial community structural shift from net autotrophic to net heterotrophic. Despite this shift, microbial grazers (mostly choreotrich ciliates, taxon-specific growth rates up to 2.9 d(-1)) controlled nanophytoplankton and bacteria at all sites by consuming 76 to 110% and 56 to 97% of their daily production, respectively, in dilution experiments. Overall, distribution patterns and dynamics of microbial communities in OWC resemble those in marine estuaries, where plankton productivity increases along the river-sea gradient and reaches its maximum at the confluence.

Integrator complex plays an essential role in adipose differentiation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Otani, Yuichiro; Nakatsu, Yusuke; Sakoda, Hideyuki

2013-05-03

Highlights: •IntS6 and IntS11 are subunits of the Integrator complex. •Expression levels of IntS6 and IntS11 were very low in 3T3-L1 fibroblast. •IntS6 and IntS11 were upregulated during adipose differentiation. •Suppression of IntS6 or IntS11 expression inhibited adipose differentiation. -- Abstract: The dynamic process of adipose differentiation involves stepwise expressions of transcription factors and proteins specific to the mature fat cell phenotype. In this study, it was revealed that expression levels of IntS6 and IntS11, subunits of the Integrator complex, were increased in 3T3-L1 cells in the period when the cells reached confluence and differentiated into adipocytes, while being reducedmore » to basal levels after the completion of differentiation. Suppression of IntS6 or IntS11 expression using siRNAs in 3T3-L1 preadipocytes markedly inhibited differentiation into mature adipocytes, based on morphological findings as well as mRNA analysis of adipocyte-specific genes such as Glut4, perilipin and Fabp4. Although Pparγ2 protein expression was suppressed in IntS6 or IntS11-siRNA treated cells, adenoviral forced expression of Pparγ2 failed to restore the capacity for differentiation into mature adipocytes. Taken together, these findings demonstrate that increased expression of Integrator complex subunits is an indispensable event in adipose differentiation. Although further study is necessary to elucidate the underlying mechanism, the processing of U1, U2 small nuclear RNAs may be involved in cell differentiation steps.« less

Are biological control agents, isolated from tropical fruits, harmless to potential consumers?

PubMed

Ocampo-Suarez, Iris Betsabee; López, Zaira; Calderón-Santoyo, Montserrat; Ragazzo-Sánchez, Juan Arturo; Knauth, Peter

2017-11-01

Postharvest losses of fruits and vegetables can reach up to 25% in developed and up to 50% in developing countries. (Sub)tropical fruits are especially susceptible because their protecting peel can easily be damaged. Traditionally used pesticides are associated to environmental pollution and possible harmful health effects. An alternative are biocontrol agents (BCA), means bacteria or yeasts applied onto the fruits to inhibit the growth of phytopathogens. Many reports on their effectiveness have been published, however, reports on their harmlessness to consumers are still rare. Culture extracts of six BCAs, tested on two human lines (Caco-2, HeLa), exhibited no cytotoxic effect, when used directly (1×) to protect the fruits; however, when they are 5×overconcentrated, the confluence of proliferating cells was reduced, but not of differentiated Caco-2. In both cases necrosis was not increased. On proliferating cells, the 5×-extract from Cryptococcus laurentii or Debaryomyces hansenii reduced lysosome functionality and the 6.25×extract from Meyerozyma guilliermondii or Candida famata increased membrane permeability, while only the 25×-extract from M. guilliermondii or M. caribbica reduced slightly the metabolic activity. The extract of Bacillus subtilis showed no cytotoxic effect up to 10× concentration. Overall, their low cytotoxicity combined with high biodegradability make these products suitable for sustainable agriculture. Copyright © 2017 Elsevier Ltd. All rights reserved.

Channel change and bed-material transport in the Lower Chetco River, Oregon

USGS Publications Warehouse

Wallick, J. Rose; Anderson, Scott W.; Cannon, Charles; O'Connor, Jim E.

2010-01-01

The lower Chetco River is a wandering gravel-bed river flanked by abundant and large gravel bars formed of coarse bed-material sediment. Since the early twentieth century, the large gravel bars have been a source of commercial aggregate for which ongoing permitting and aquatic habitat concerns have motivated this assessment of historical channel change and sediment transport rates. Analysis of historical channel change and bed-material transport rates for the lower 18 kilometers shows that the upper reaches of the study area are primarily transport zones, with bar positions fixed by valley geometry and active bars mainly providing transient storage of bed material. Downstream reaches, especially near the confluence of the North Fork Chetco River, are zones of active sedimentation and channel migration.Multiple analyses, supported by direct measurements of bedload during winter 2008–09, indicate that since 1970 the mean annual flux of bed material into the study reach has been about 40,000–100,000 cubic meters per year. Downstream tributary input of bed-material sediment, probably averaging 5–30 percent of the influx coming into the study reach from upstream, is approximately balanced by bed-material attrition by abrasion. Probably little bed material leaves the lower river under natural conditions, with most net influx historically accumulating in wider and more dynamic reaches, especially near the North Fork Chetco River confluence, 8 kilometers upstream from the Pacific Ocean.The year-to-year flux, however, varies tremendously. Some years may have less than 3,000 cubic meters of bed material entering the study area; by contrast, some high-flow years, such as 1982 and 1997, likely have more than 150,000 cubic meters entering the reach. For comparison, the estimated annual volume of gravel extracted from the lower Chetco River for commercial aggregate during 2000–2008 has ranged from 32,000 to 90,000 cubic meters and averaged about 59,000 cubic meters per year. Mined volumes probably exceeded 140,000 cubic meters per year for several years in the late 1970s.Repeat surveys and map analyses indicate a reduction in bar area and sinuosity between 1939 and 2008, chiefly in the period 1965–95. Repeat topographic and bathymetric surveys show channel incision for substantial portions of the study reach, with local areas of bed lowering by as much as 2 meters. A specific gage analysis at the upstream end of the study reach indicates that incision and narrowing followed aggradation culminating in the late 1970s. These observations are all consistent with a reduction of sediment supply relative to transport capacity since channel surveys in the late 1970s, probably owing to a combination of (1) bed sediment removal and (2) transient river adjustments to large sediment volumes brought by floods such as those in 1964 and, to a lesser extent, 1996.

Channel change and bed-material transport in the Lower Chetco River, Oregon

USGS Publications Warehouse

Wallick, J. Rose; Anderson, Scott W.; Cannon, Charles; O'Connor, Jim E.

2009-01-01

The lower Chetco River is a wandering gravel-bed river flanked by abundant and large gravel bars formed of coarse bed-material sediment. The large gravel bars have been a source of commercial aggregate since the early twentieth century for which ongoing permitting and aquatic habitat concerns have motivated this assessment of historical channel change and sediment transport rates. Analysis of historical channel change and bed-material transport rates for the lower 18 kilometers show that the upper reaches of the study area are primarily transport zones, with bar positions fixed by valley geometry and active bars mainly providing transient storage of bed material. Downstream reaches, especially near the confluence of the North Fork Chetco River, have been zones of active sedimentation and channel migration.Multiple analyses, supported by direct measurements of bedload during winter 2008–09, indicate that since 1970 the mean annual flux of bed material into the study reach has been about 40,000–100,000 cubic meters per year. Downstream tributary input of bed-material sediment, probably averaging 5–30 percent of the influx coming into the study reach from upstream, is approximately balanced by bed-material attrition by abrasion. Probably very little bed material leaves the lower river under natural conditions, with most of the net influx historically accumulating in wider and more dynamic reaches, especially near the North Fork Chetco River confluence, 8 kilometers upstream from the Pacific Ocean.The year-to-year flux, however, varies tremendously. Some years probably have less than 3,000 cubic meters of bed-material entering the study area; by contrast, some high-flow years, such as 1982 and 1997, likely have more than 150,000 cubic meters entering the reach. For comparison, the estimated annual volume of gravel extracted from the lower Chetco River for commercial aggregate during 2000–2008 has ranged from 32,000 to 90,000 cubic meters and averaged about 59,000 cubic meters per year. Mined volumes probably exceeded 140,000 cubic meters per year for several years in the late 1970s.Repeat surveys and map analyses indicate a reduction in bar area and sinuosity between 1939 and 2008, chiefly in the period 1965–95. Repeat topographic and bathymetric surveys show channel incision for substantial portions of the study reach, with local areas of bed lowering by as much as 2 meters. A specific gage analysis at the upstream end of the study reach indicates that incision and narrowing followed aggradation culminating in the late 1970s. These observations are all consistent with a reduction of sediment supply relative to transport capacity since channel surveys in the late 1970s, probably owing to a combination of (1) bed-sediment removal and (2) transient river adjustments to large sediment volumes brought by floods such as those in 1964, and to a lesser extent, 1996.

Channel Change and Bed-Material Transport in the Lower Chetco River, Oregon

NASA Astrophysics Data System (ADS)

O'Connor, J. E.; Wallick, R.; Anderson, S.; Cannon, C.

2009-12-01

The Chetco River drains 914 square kilometers of the Klamath Mountains in far southwestern Oregon. For its lowermost 18 km, it is a wandering gravel-bed river flanked by abundant and large gravel bars formed of coarse bed-material sediment. The large gravel bars have been a source of commercial aggregate since the early twentieth century for which ongoing permitting and aquatic habitat concerns have motivated an assessment of historical channel change and sediment transport rates. Analysis of historical channel change and bed-material transport rates for the lower 18 kilometers show that the upper reaches of the study area are primarily transport zones, with bar positions fixed by valley geometry and active bars mainly providing transient storage of bed material. Downstream reaches, especially near the confluence of the North Fork Chetco River, have been zones of active sedimentation and channel migration. Multiple analyses, supported by direct measurements of bedload during winter 2008-09, indicate that since 1970 the mean annual flux of bed material into the study reach has been about 40,000-100,000 cubic meters per year. Downstream tributary input of bed-material sediment, probably averaging 5-30 percent of the influx coming into the study reach from upstream, is approximately balanced by bed-material attrition by abrasion. Probably very little bed material leaves the lower river under natural conditions, with most of the net influx historically accumulating in wider and more dynamic reaches, especially near the North Fork Chetco River confluence, 8 kilometers upstream from the Pacific Ocean. The year-to-year flux, however, varies tremendously. Some years probably have less than 3,000 cubic meters of bed-material entering the study area; by contrast, some high-flow years, such as 1982 and 1997, likely have more than 150,000 cubic meters entering the reach. For comparison, the estimated annual volume of gravel extracted from the lower Chetco River for commercial aggregate during 2000-2008 has ranged from 32,000 to 90,000 cubic meters and averaged about 59,000 cubic meters per year. Mined volumes probably exceeded 140,000 cubic meters per year for several years in the late 1970s. Repeat surveys and map analyses indicate a reduction in bar area and sinuosity between 1939 and 2008, chiefly in the period 1965-95. Repeat topographic and bathymetric surveys show channel incision for substantial portions of the study reach, with local areas of bed lowering by as much as 2 meters. A specific gage analysis at the upstream end of the study reach indicates that incision and narrowing followed aggradation culminating in the late 1970s. These observations are all consistent with a reduction of sediment supply relative to transport capacity since channel surveys in the late 1970s, probably owing to a combination of (1) bed-sediment removal and (2) transient river adjustments to large sediment volumes brought by floods such as those in 1964, and to a lesser extent, 1996.

Field Investigation of Flow Structure and Channel Morphology at Confluent-Meander Bends

NASA Astrophysics Data System (ADS)

Riley, J. D.; Rhoads, B. L.

2007-12-01

The movement of water and sediment through drainage networks is inevitably influenced by the convergence of streams and rivers at channel confluences. These focal components of fluvial systems produce a complex hydrodynamic environment, where rapid changes in flow structure and sediment transport occur to accommodate the merging of separate channel flows. The inherent geometric and hydraulic change at confluences also initiates the development of distinct geomorphic features, reflected in the bedform and shape of the channel. An underlying assumption of previous experimental and theoretical models of confluence dynamics has been that converging streams have straight channels with angular configurations. This generalized conceptualization was necessary to establish confluence planform as symmetrical or asymmetrical and to describe subsequent flow structure and geomorphic features at confluences. However, natural channels, particularly those of meandering rivers, curve and bend. This property and observation of channel curvature at natural junctions have led to the hypothesis that natural stream and river confluences tend to occur on the concave outer bank of meander bends. The resulting confluence planform, referred to as a confluent-meander bend, was observed over a century ago but has received little scientific attention. This paper examines preliminary data on three-dimensional flow structure and channel morphology at two natural confluent-meander bends of varying size and with differing tributary entrance locations. The large river confluence of the Vermilion River and Wabash River in west central Indiana and the comparatively small junction of the Little Wabash River and Big Muddy Creek in southeastern Illinois are the location of study sites for field investigation. Measurements of time-averaged three-dimensional velocity components were obtained at these confluences with an acoustic Doppler current profiler for flow events with differing momentum ratios. Bed and channel morphology were also surveyed with a digital fathometer to document geomorphic change. Preliminary analysis of the velocity data reveals the presence of a well-defined shear layer between the converging flows and secondary circulation in the main channel. The tributary channel appears to oppose high velocity flow directed toward the outer bank by centrifugal acceleration through the meander bend of the main channel, thereby diminishing erosion along the cut bank and possibly stabilizing the meander bend channel. The flow structure and channel morphology of the study sites are compared to consider the effect of spatial scale and geometric characteristics on confluent-meander bend dynamics.

Use of multispectral satellite remote sensing to assess mixing of suspended sediment downstream of large river confluences

NASA Astrophysics Data System (ADS)

Umar, M.; Rhoads, Bruce L.; Greenberg, Jonathan A.

2018-01-01

Although past work has noted that contrasts in turbidity often are detectable on remotely sensed images of rivers downstream from confluences, no systematic methodology has been developed for assessing mixing over distance of confluent flows with differing surficial suspended sediment concentrations (SSSC). In contrast to field measurements of mixing below confluences, satellite remote-sensing can provide detailed information on spatial distributions of SSSC over long distances. This paper presents a methodology that uses remote-sensing data to estimate spatial patterns of SSSC downstream of confluences along large rivers and to determine changes in the amount of mixing over distance from confluences. The method develops a calibrated Random Forest (RF) model by relating training SSSC data from river gaging stations to derived spectral indices for the pixels corresponding to gaging-station locations. The calibrated model is then used to predict SSSC values for every river pixel in a remotely sensed image, which provides the basis for mapping of spatial variability in SSSCs along the river. The pixel data are used to estimate average surficial values of SSSC at cross sections spaced uniformly along the river. Based on the cross-section data, a mixing metric is computed for each cross section. The spatial pattern of change in this metric over distance can be used to define rates and length scales of surficial mixing of suspended sediment downstream of a confluence. This type of information is useful for exploring the potential influence of various controlling factors on mixing downstream of confluences, for evaluating how mixing in a river system varies over time and space, and for determining how these variations influence water quality and ecological conditions along the river.

77 FR 76916 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2012-12-31

... Southeast. Sweat Mountain Creek At the Willeo Creek +941 Unincorporated Areas of confluence. Cobb County... upstream of the Sweat Mountain Creek confluence. * National Geodetic Vertical Datum. + North American...

Hydrographic survey of Chaktomuk, the confluence of the Mekong, Tonlé Sap, and Bassac Rivers near Phnom Penh, Cambodia, 2012

USGS Publications Warehouse

Dietsch, Benjamin J.; Densmore, Brenda K.; Wilson, Richard C.

2014-01-01

Detailed hydrographic maps of Mekong, Tonlé Sap, and Bassac Rivers showing the riverbed elevations surveyed April 21–May 2, 2012, referenced to Ha Tien 1960 were produced. The surveyed area included a 2-km stretch of the Mekong River between the confluence with the Tonlé Sap and Bassac Rivers, and extended 4 km upstream and 3.6 km downstream from the 2,000-m confluence stretch of the Mekong River. In addition, 0.7 km of the Bassac River downstream and 3.5 km of the Tonlé Sap River (from the confluence to Chroy Changvar Bridge) upstream from their confluence with the Mekong River were surveyed. Riverbed features (such as dunes, shoals, and the effects of sediment mining, which were observed during data collection) are visible on the hydrographic maps. All surveys were completed at low annual water levels as referenced to nearby Mekong River Commission streamflow-gaging stations. Riverbed elevations surveyed ranged from 24.08 m below to 1.54 m above Ha Tien 1960.

Morphological changes and viability of primary cultured human ocular trabecular meshwork cells after exposure to air.

PubMed

Kopsachilis, Nikolaos; Tsaousis, Konstantinos T; Carifi, Gianluca; Welge-Luessen, Ulrich

2014-06-01

To investigate the possible toxic effect of air exposure for an in vitro model of primary human ocular trabecular meshwork cells (HTM). HTM were isolated from five donor eyes and cultivated at 37 °C. After reaching confluence the cells were seeded on two well chamber slides. The chamber slides were turned upside down in a Petri culture dish full of culture medium and filled with air using a 5 ml syringe, starting this way the exposure of the cells to the air. Subsequently they were placed in the incubation chamber at 37 °C. Six groups of HTM cultures were set up: group 1 consisted of samples in which HTM were exposed to air for 30 min, group 2 for 1 h, group 3 for 3 h, group 4 for 6 h, group 5 for 12 h and group 6 for 24 h. At 3 h after exposure, the morphology of the cells was still intact, at 6 h few cells appeared deformed and exhibited characteristics of more senescent cells. At 12 h after exposure to air the HTM cells started losing their typical morphology and appeared enlarged and compromised. Viability was superior to 94% in groups 1-3 while for groups 4, 5, 6 it was 82.7%, 39.5% and 12.7% respectively. The toxic effect of air exposure for the studied in vitro model of HTM is not significant for the time period of one to three hours. However it starts reducing viability and alternating morphology 6 h after exposure until the time period of 24 h, where the percentage of living cells is drastically decreased. Therefore, we suggest that the use of an air bubble especially in glaucomatous patients should be applied with caution. Copyright © 2014 Elsevier Ltd. All rights reserved.

Water quality of the Delaware and Raritan Canal, New Jersey, 1998-99

USGS Publications Warehouse

Gibs, Jacob; Gray, Bonnie; Rice, Donald E.; Tessler, Steven; Barringer, Thomas H.

2001-01-01

The mean and median of continuously monitored turbidity varied along the length of the canal. In the reach between Raven Rock and Lower Ferry Road, the mean and median for continuously monitored turbidity during the study period increased by 7.2 and 6.2 NTU (nephelometric turbidity units), respectively. The mean of continuously monitored turbidity decreased downstream from Lower Ferry Road to Ten Mile Lock. Turbidity could increase locally downstream from influent streams or outfalls, but because the average velocity of water in the canal is low, particles that cause turbidity are not transported appreciable distances. In the reach between Ten Mile Lock and the Route 18 Spillway, the mean and median of the continuously monitored turbidity changed less than 0.5 NTU during the period of record. The small change in turbidity in this reach is not consistent with an average velocity for the reach; the average velocity in this reach was the lowest in all of the reaches studied. The expected decrease in turbidity due to settling of suspended solids is likely offset by turbid water entering the canal from influent streams or discharges from storm drains. Field observation of a sand bar immediately downstream from the confluence of Als Brook and the canal confirmed that the Als Brook drainage basin has contributed stormwatergenerated sediment to the canal that could reach the monitor located at the Route 18 Spillway and the raw water intakes for two drinking-water treatment plants.

A joint probability approach for coincidental flood frequency analysis at ungauged basin confluences

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Cheng

2016-03-12

A reliable and accurate flood frequency analysis at the confluence of streams is of importance. Given that long-term peak flow observations are often unavailable at tributary confluences, at a practical level, this paper presents a joint probability approach (JPA) to address the coincidental flood frequency analysis at the ungauged confluence of two streams based on the flow rate data from the upstream tributaries. One case study is performed for comparison against several traditional approaches, including the position-plotting formula, the univariate flood frequency analysis, and the National Flood Frequency Program developed by US Geological Survey. It shows that the results generatedmore » by the JPA approach agree well with the floods estimated by the plotting position and univariate flood frequency analysis based on the observation data.« less

The morphodynamics and sedimentology of large river confluences

NASA Astrophysics Data System (ADS)

Nicholas, Andrew; Sambrook Smith, Greg; Best, James; Bull, Jon; Dixon, Simon; Goodbred, Steven; Sarker, Mamin; Vardy, Mark

2017-04-01

Confluences are key locations within large river networks, yet surprisingly little is known about how they migrate and evolve through time. Moreover, because confluence sites are associated with scour pools that are typically several times the mean channel depth, the deposits associated with such scours should have a high potential for preservation within the rock record. However, paradoxically, such scours are rarely observed, and the sedimentological characteristics of such deposits are poorly understood. This study reports results from a physically-based morphodynamic model, which is applied to simulate the evolution and resulting alluvial architecture associated with large river junctions. Boundary conditions within the model simulation are defined to approximate the junction of the Ganges and Jamuna rivers, in Bangladesh. Model results are supplemented by geophysical datasets collected during boat-based surveys at this junction. Simulated deposit characteristics and geophysical datasets are compared with three existing and contrasting conceptual models that have been proposed to represent the sedimentary architecture of confluence scours. Results illustrate that existing conceptual models may be overly simplistic, although elements of each of the three conceptual models are evident in the deposits generated by the numerical simulation. The latter are characterised by several distinct styles of sedimentary fill, which can be linked to particular morphodynamic behaviours. However, the preserved characteristics of simulated confluence deposits vary substantial according to the degree of reworking by channel migration. This may go some way towards explaining the confluence scour paradox; while abundant large scours might be expected in the rock record, they are rarely reported.

Independent cellular effects of cold ischemia and reperfusion: experimental molecular study.

PubMed

Lledó-García, E; Humanes-Sánchez, B; Mojena-Sánchez, M; Rodrígez, J C J; Hernández-Fernández, C; Tejedor-Jorge, A; Fernández, A L

2013-04-01

There is less information available on cell cultures on the exclusive effects of either duration of cold ischemia (CI) or rewarming-reperfusion in the kidney subjected to initial warm ischemia (WI). Therefore, the goals of our work were: (1) to evaluate the consequences on tubular cellular viability of different durations of CI on a kidney after an initial period of WI, and (2) to analyze the additional effect on tubular cell viability of rewarming of the same kidney. Sixteen mini-pig were used. All the animals were performed a right nephrectomy after 45-minute occlusion of the vascular pedicle. The kidneys were then divided into 2 groups (phase 1): cold storage in university of wisconsin (UW) solution for 3 hours (group A, n = 8) at 4°C, or cold storage in UW for 12 hours (group B, n = 8) at 4°C. Four organs of group A and four organs of group B were autotrasplanted (AT) and reperfused for 1 hour (phase 2). Nephrectomy was finally done. Biopsies were taken from all groups to perform cultures of proximal tubule epithelium cells. The biopsies were subjected to studies of cellular morphological viability (contrast phase microscopy [CPM]) and quantitative (confluence cell [CC]) parameters. Phase of pure CI effects (phase 1): Both CC rate and CPM parameters were significantly lower in group B compared with group A, where cell activity reached almost normal results. Phase of CI + AT (phase 2): At produced additional harmful effects in cell cultures compared with those obtained in phase 1, more evident in group B cells. The presence of cold storage followed by rewarming-reperfusion induces independent and cumulative detrimental effects in viability of renal proximal tubule cells. CI periods ≤ 3 hours may ameliorate the injuries secondary to reperfusion in comparison with longer CI periods. Copyright © 2013 Elsevier Inc. All rights reserved.

Characterization of corneal pannus removed from patients with total limbal stem cell deficiency.

PubMed

Espana, Edgar M; Di Pascuale, Mario A; He, Hua; Kawakita, Tetsuya; Raju, Vadrevu K; Liu, Chia-Yang; Tseng, Scheffer C G

2004-09-01

To determine the epithelial lineage of origin in corneal pannus tissue surgically removed from patients with total limbal stem cell (SC) deficiency. The lineage of origin of the entire conjunctivalized pannus removed from eight corneas with a diagnosis of total limbal SC deficiency was characterized by anti-keratin (K)-3 and anti-K19 monoclonal antibodies. The protein and mRNA of epithelial outgrowth from segments of five such pannus specimens were analyzed by Western blot and reverse transcription-polymerase chain reaction, respectively. Cross sections of all eight specimens showed a stratified epithelium with goblet cells expressing mucin (MUC)-5AC, and a stroma showing blood vessels and inflammatory cell infiltrates. Immunostaining showed full-thickness expression of K19 in the entire pannus of all eight specimens. Expression of K3 was negative in seven patients, but was sporadically positive in a patient with Stevens-Johnson syndrome. In culture, all five pannus specimens generated a compact, small epithelial cell outgrowth, and except for one, reached confluence in 2 to 3 weeks. The K3/K12 pair was expressed by extracts of cell outgrowth from the control limbal epithelial explant, but not in all five pannus specimens. A 60-kDa band of DeltaNp63 was expressed in the control specimen and in all five pannus specimens. Cell outgrowth expressed K3 transcript in three, but none showed K12 transcript. The resultant epithelial phenotype of the pannus tissue was not corneal, as evidenced by the negative staining to cornea-specific K12 mRNA and protein, but was conjunctival, as evidenced by the presence of goblet cells, the weak expression of K3, and the strong expression of K19. The abundant expression of DeltaNp63 in such conjunctiva-derived epithelium in eyes with total limbal SC deficiency raises doubts as to its validity as a limbal SC marker. Copyright Association for Research in Vision and Ophthalmology

Contributions of moderately low flows and large floods to geomorphic change in the Rio Puerco Arroyo, New Mexico

USGS Publications Warehouse

Griffin, Eleanor R.; Friedman, Jonathan M.

2016-01-01

Abstract—From the mid-1800s to around 1930, monsoonal floods incised an arroyo roughly 100 m wide and 10 m deep along the lower Rio Puerco, NM, from the confluence with the Rio San Jose downstream to the mouth at the Rio Grande, causing sedimentation and flooding downstream. Since the 1930s, the channel has greatly narrowed, a densely vegetated floodplain has developed, the arroyo has partly filled, and downstream sedimentation has greatly decreased. Application of herbicide to a 12-km reach of the arroyo in 2003 to control non-native saltcedar (Tamarix spp.) prompted ongoing studies of channel change in the presence and absence of dense, riparian, woody vegetation. We used digital terrain models and satellite imagery to quantify changes in channel width and location in the sprayed reach and in an unsprayed reach downstream during a moderately low-flow interval (November 2006 to March 2010) and during an interval with a large flood (March 2010 to January/February 2014). Channel width increased in magnitude and variability in the sprayed reach but not in the unsprayed reach over both intervals, continuing a pattern first observed in an earlier study of the period 2003 to 2006. Since the herbicide application in 2003, there have been a total of five meander cutoffs in the sprayed reach and none in the unsprayed reach. In kilometer-long sections of the sprayed reach, channel width is now approaching that at the beginning of the period of channel narrowing in 1935.

A preliminary assessment of streamflow gains and losses for selected stream reaches in the lower Guadalupe River Basin, Texas, 2010-12

USGS Publications Warehouse

Wehmeyer, Loren L.; Winters, Karl E.; Ockerman, Darwin J.

2013-01-01

During the August 19–25, 2011, base-flow period, three reaches had gains greater than the uncertainty in the computed streamflow, including reach 3 on the Comal River (168 ft3/s gain), which was one of the reaches where gains in streamflow also were measured in March 2010 and April 2011. Streamflow gains in August 2011 were primarily from (1) inflows from Comal Springs, (2) inflows from the Yegua Jackson aquifer, and (3) groundwater inflows from the Gulf Coast aquifer, which are enhanced by seepage losses from Coleto Creek Reservoir. During this base-flow period, five reaches had losses greater in magnitude than the uncertainty in the computed streamflow. The reach including the confluence of the Guadalupe and Comal Rivers lost 82.8 ft3/s. Much of that loss likely seeped into the local groundwater system. The reach of the Guadalupe River south of New Braunfels, Tex., to Seguin, Tex., lost 53.5 ft3/s. Part of that loss may have been from seepage through streambed alluvium. Reaches 9 and 10 of the Blanco River near Kyle lost 2.20 and 6.60 ft3/s, respectively, likely as infiltration through numerous faults intersecting the stream channel northwest of Kyle. Plum Creek between Lockhart, Tex., and Luling, Tex., lost 2.11 ft3/s, likely as recharge to the Carrizo-Wilcox aquifer. A base-flow period during September 22–28, 2012, was studied for the reach of the Guadalupe River between Seguin and Gonzalez, including flows from San Marcos River and Plum Creek. During this period, for the Guadalupe River reach between Seguin and Oak Forest, no computed gains or losses were greater in magnitude than the uncertainty in the computed streamflow.

MicroRNA expression profile and functional analysis reveal their roles in contact inhibition and its disruption switch of rat vascular smooth muscle cells.

PubMed

Sun, Ye-Ying; Qin, Shan-Shan; Cheng, Yun-Hui; Wang, Chao-Yun; Liu, Xiao-Jun; Liu, Ying; Zhang, Xiu-Li; Zhang, Wendy; Zhan, Jia-Xin; Shao, Shuai; Bian, Wei-Hua; Luo, Bi-Hui; Lu, Dong-Feng; Yang, Jian; Wang, Chun-Hua; Zhang, Chun-Xiang

2018-05-01

Contact inhibition and its disruption of vascular smooth muscle cells (VSMCs) are important cellular events in vascular diseases. But the underlying molecular mechanisms are unclear. In this study we investigated the roles of microRNAs (miRNAs) in the contact inhibition and its disruption of VSMCs and the molecular mechanisms involved. Rat VSMCs were seeded at 30% or 90% confluence. MiRNA expression profiles in contact-inhibited confluent VSMCs (90% confluence) and non-contact-inhibited low-density VSMCs (30% confluence) were determined. We found that multiple miRNAs were differentially expressed between the two groups. Among them, miR-145 was significantly increased in contact-inhibited VSMCs. Serum could disrupt the contact inhibition as shown by the elicited proliferation of confluent VSMCs. The contact inhibition disruption accompanied with a down-regulation of miR-145. Serum-induced contact inhibition disruption of VSMCs was blocked by overexpression of miR-145. Moreover, downregulation of miR-145 was sufficient to disrupt the contact inhibition of VSMCs. The downregulation of miR-145 in serum-induced contact inhibition disruption was related to the activation PI3-kinase/Akt pathway, which was blocked by the PI3-kinase inhibitor LY294002. KLF5, a target gene of miR-145, was identified to be involved in miR-145-mediated effect on VSMC contact inhibition disruption, as it could be inhibited by knockdown of KLF5. In summary, our results show that multiple miRNAs are differentially expressed in contact-inhibited VSMCs and in non-contact-inhibited VSMCs. Among them, miR-145 is a critical gene in contact inhibition and its disruption of VSMCs. PI3-kinase/Akt/miR-145/KLF5 is a critical signaling pathway in serum-induced contact inhibition disruption. Targeting of miRNAs related to the contact inhibition of VSMCs may represent a novel therapeutic approach for vascular diseases.

Population ecology of variegate darter (Etheostoma variatum) in Virginia

USGS Publications Warehouse

Argentina, Jane E.; Angermeier, Paul; Hallerman, Eric M.

2013-01-01

Variegate darters (Etheostoma variatum) were listed as endangered in Virginia in 1992. Reasons for listing included habitat degradation and concerns about current and future impacts of coal mining throughout their Virginia range. Prior to this research, little was known about variegate darter distribution, habitat use, or populations in Virginia. Two primary goals of this research were to gain knowledge about the current population ecology and the relationship between landscape-level factors (e.g., land cover changes, watershed size, isolation from other populations) on current and past variegate darter population sizes.We investigated distribution, habitat suitability, population genetics, and population size and structure of variegate darters in the upper Big Sandy River drainage, Buchanan, Dickenson, and Wise Co., Virginia. Our results indicate variegate darters are primarily found in the Levisa Fork, with highest densities and abundances between its confluence with Dismal Creek and the Virginia-Kentucky border. Sporadic occurrences in smaller tributaries to the Levisa and Tug forks indicate they exist more widely in low densities, especially near the confluence with the Tug and Levisa mainstems. Detection of variegate darters in smaller tributaries was inconsistent, with reach-level occupancy estimates varying among years. We detected young-of-year variegate darters every year we sampled, but age 1+ darters were indistinguishable from older darters based on standard length.Variegate darter population size and stability in Virginia were estimated via multiple methods, including site occupancy surveys, mark-recapture studies, and population genetic analysis. Using mark-recapture methods at five sites, we estimated overall population size in 2011 to be approximately 12,800 individuals in the 35-km reach between the Levisa Fork - Dismal Creek confluence and the Virginia-Kentucky border. Age structure seemed stable, with breeding adults and young-of-year collected annually during 2008-2011. Population genetic analysis indicated variegate darters in the Levisa Fork and its tributaries are part of a single genetic population. Historical and current genetic stability were seen in our analysis of the variegate darter population, with no genetic differentiation among riffles across the upper Levisa Fork watershed, indicating dispersal among these sites is enough to overcome random genetic drift. This population is genetically isolated from downstream populations by the dam at Fishtrap Lake, Pike Co., Kentucky, and is beginning to show genetic isolation from other nearby populations. As expected, the Virginia population is most closely related to those in the Russell Fork and Levisa Fork downstream of the dam.Regular monitoring of variegate darters in the Levisa Fork mainstem from the Dismal Creek confluence to the Virginia-Kentucky border would facilitate better understanding of normal fluctuations of population size and distribution, as well as assessments of population status. This reach encompasses the core of the variegate darter population in Virginia, and its persistence will determine long-term viability of this species. Given that little is known about long-term population trends, we suggest that annual site-occupancy and population size estimates be made at ten randomly selected riffles for at least ten years to understand normal levels of variability. Thereafter, these population parameters could be monitored bi-annually as a way to detect shrinking distribution or abundance, especially after any fish kill or other pollution event in the Levisa Fork. We further suggest that the sites upstream and downstream of the saline diffusor pipe be monitored to detect changes in the extent of the impact zone.Overall, the variegate darter population in Virginia appears stable, although primarily confined to the lower 35 km of the Levisa Fork. Nevertheless, variegate darters in Virginia remain susceptible to extirpation due to catastrophic events, both physical (chemical spill) and biological (disease outbreak or invasive species introduction).

The Nucleus Introduced

PubMed Central

Pederson, Thoru

2011-01-01

Now is an opportune moment to address the confluence of cell biological form and function that is the nucleus. Its arrival is especially timely because the recognition that the nucleus is extremely dynamic has now been solidly established as a paradigm shift over the past two decades, and also because we now see on the horizon numerous ways in which organization itself, including gene location and possibly self-organizing bodies, underlies nuclear functions. PMID:20660024

Effects of Glen Canyon Dam discharges on water velocity and temperatures at the confluence of the Colorado and Little Colorado Rivers and implications for habitat for young-of-year humpback chub (Gila cypha-

USGS Publications Warehouse

Protiva, Frank R.; Ralston, Barbara E.; Stone, Dennis M.; Kohl, Keith A.; Yard, Michael D.; Haden, G. Allen

2010-01-01

Water velocity and temperature are physical variables that affect the growth and survivorship of young-of-year (YOY) fishes. The Little Colorado River, a tributary to the Colorado River in Grand Canyon, is an important spawning ground and warmwater refuge for the endangered humpback chub (Gila cypha) from the colder mainstem Colorado River that is regulated by Glen Canyon Dam. The confluence area of the Little Colorado River and the Colorado River is a site where YOY humpback chub (size 30-90 mm) emerging from the Little Colorado River experience both colder temperatures and higher velocities associated with higher mainstem discharge. We used detailed surveying and mapping techniques in combination with YOY velocity and temperature preferenda (determined from field and lab studies) to compare the areal extent of available habitat for young fishes at the confluence area under four mainstem discharges (227, 368, 504, and 878 m3/s). Comparisons revealed that the areal extent of low-velocity, warm water at the confluence decreased when discharges exceeded 368 m3/s. Furthermore, mainstem fluctuations, depending on the rate of upramp, can affect velocity and temperature dynamics in the confluence area within several hours. The amount of daily fluctuations in discharge can result in the loss of approximately 1.8 hectares of habitat favorable to YOY humpback chub. Consequently, flow fluctuations and the accompanying changes in velocity and temperature at the confluence may diminish the recruitment potential of humpback chub that spawn in the tributary stream. This study illustrates the utility of multiple georeferenced data sources to provide critical information related to the influence of the timing and magnitude of discharge from Glen Canyon Dam on potential rearing environment at the confluence area of the Little Colorado River.

Desertification, salinization, and biotic homogenization in a dryland river ecosystem

USGS Publications Warehouse

Miyazono, S.; Patino, Reynaldo; Taylor, C.M.

2015-01-01

This study determined long-term changes in fish assemblages, river discharge, salinity, and local precipitation, and examined hydrological drivers of biotic homogenization in a dryland river ecosystem, the Trans-Pecos region of the Rio Grande/Rio Bravo del Norte (USA/Mexico). Historical (1977-1989) and current (2010-2011) fish assemblages were analyzed by rarefaction analysis (species richness), nonmetric multidimensional scaling (composition/variability), multiresponse permutation procedures (composition), and paired t-test (variability). Trends in hydrological conditions (1970s-2010s) were examined by Kendall tau and quantile regression, and associations between streamfiow and specific conductance (salinity) by generalized linear models. Since the 1970s, species richness and variability of fish assemblages decreased in the Rio Grande below the confluence with the Rio Conchos (Mexico), a major tributary, but not above it. There was increased representation of lower-flow/higher-salinity tolerant species, thus making fish communities below the confluence taxonomically and functionally more homogeneous to those above it. Unlike findings elsewhere, this biotic homogenization was due primarily to changes in the relative abundances of native species. While Rio Conchos discharge was > 2-fold higher than Rio Grande discharge above their confluence, Rio Conchos discharge decreased during the study period causing Rio Grande discharge below the confluence to also decrease. Rio Conchos salinity is lower than Rio Grande salinity above their confluence and, as Rio Conchos discharge decreased, it caused Rio Grande salinity below the confluence to increase (reduced dilution). Trends in discharge did not correspond to trends in precipitation except at extreme-high (90th quantile) levels. In conclusion, decreasing discharge from the Rio Conchos has led to decreasing flow and increasing salinity in the Rio Grande below the confluence. This spatially uneven desertification and salinization of the Rio Grande has in turn led to a region-wide homogenization of hydrological conditions and of taxonomic and functional attributes of fish assemblages.

Desertification, salinization, and biotic homogenization in a dryland river ecosystem.

PubMed

Miyazono, Seiji; Patiño, Reynaldo; Taylor, Christopher M

2015-04-01

This study determined long-term changes in fish assemblages, river discharge, salinity, and local precipitation, and examined hydrological drivers of biotic homogenization in a dryland river ecosystem, the Trans-Pecos region of the Rio Grande/Rio Bravo del Norte (USA/Mexico). Historical (1977-1989) and current (2010-2011) fish assemblages were analyzed by rarefaction analysis (species richness), nonmetric multidimensional scaling (composition/variability), multiresponse permutation procedures (composition), and paired t-test (variability). Trends in hydrological conditions (1970s-2010s) were examined by Kendall tau and quantile regression, and associations between streamflow and specific conductance (salinity) by generalized linear models. Since the 1970s, species richness and variability of fish assemblages decreased in the Rio Grande below the confluence with the Rio Conchos (Mexico), a major tributary, but not above it. There was increased representation of lower-flow/higher-salinity tolerant species, thus making fish communities below the confluence taxonomically and functionally more homogeneous to those above it. Unlike findings elsewhere, this biotic homogenization was due primarily to changes in the relative abundances of native species. While Rio Conchos discharge was>2-fold higher than Rio Grande discharge above their confluence, Rio Conchos discharge decreased during the study period causing Rio Grande discharge below the confluence to also decrease. Rio Conchos salinity is lower than Rio Grande salinity above their confluence and, as Rio Conchos discharge decreased, it caused Rio Grande salinity below the confluence to increase (reduced dilution). Trends in discharge did not correspond to trends in precipitation except at extreme-high (90th quantile) levels. In conclusion, decreasing discharge from the Rio Conchos has led to decreasing flow and increasing salinity in the Rio Grande below the confluence. This spatially uneven desertification and salinization of the Rio Grande has in turn led to a region-wide homogenization of hydrological conditions and of taxonomic and functional attributes of fish assemblages. Copyright © 2015 Elsevier B.V. All rights reserved.

LOOKING EASTSOUTHEAST. Showing downstream side of completed bridge, from confluence ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

LOOKING EAST-SOUTHEAST. Showing downstream side of completed bridge, from confluence of Trinity and South Fork Trinity Rivers - South Fork Trinity River Bridge, State Highway 299 spanning South Fork Trinity River, Salyer, Trinity County, CA

1. FIRST SECTION OF PIPELINE BETWEEN CONFLUENCE POOL AND FISH ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

1. FIRST SECTION OF PIPELINE BETWEEN CONFLUENCE POOL AND FISH SCREEN. NOTE RETAINING WALL BESIDE PIPE. VIEW TO NORTH-NORTHEAST. - Santa Ana River Hydroelectric System, Pipeline to Fish Screen, Redlands, San Bernardino County, CA

The ultrastructure of imaginal disc cells in primary cultures and during cell aggregation in continuous cell lines.

PubMed

Peel, D J; Johnson, S A; Milner, M J

1990-01-01

We have examined the ultrastructure of cellular vesicles in primary cultures of wing imaginal disc cells of Drosophila melanogaster. These cells maintain the apico-basal polarity characteristic of epithelial cells. The apical surfaces secrete extracellular material into the lumen of the vesicle from plasma membrane plaques at the tip of microvilli. During the course of one passage, cells from the established cell lines grow to confluence and then aggregate into discrete condensations joined by aligned bridges of cells. Cells in these aggregates are tightly packed, and there appears to be a loss of the epithelial polarity characteristic of the vesicle cells. Elongated cell extensions containing numerous microtubules are found in aggregates, and we suggest that these may be epithelial feet involved in the aggregation process. Virus particles are commonly found both within the nucleus and the cytoplasm of cells in the aggregates.

Torcular Herophili classification and evaluation of dural venous sinus variations using digital subtraction angiography and magnetic resonance venographies.

PubMed

Gökçe, Erkan; Pınarbaşılı, Tansu; Acu, Berat; Fırat, M Murat; Erkorkmaz, Ünal

2014-08-01

The configurations of cerebral veins and dural venous sinuses differ not only between individuals, but also between the two brain hemispheres of an individual, making the anatomical classification of the cerebral veins difficult. We evaluated the superior dural venous sinuses and classified their types and variations using magnetic resonance venography (MRV) and digital substraction angiography (DSA). A total of 394 patients were studied retrospectively. Superior dural venous sinuses were evaluated and the confluence of the sinuses was classified on 2-dimensional time-of-flight MRV, contrast-enhanced 3-dimensional spoiled gradient recalled echo magnetic resonance imaging, and/or cerebral DSA. Confluens sinuum was divided into three types: true confluence, partial confluence, and non-confluence. Of the three types, partial confluence (type II) was most frequently seen. Co-dominance of the transverse sinuses was most frequently observed. An occipital sinus was observed in 15 % of the patients. There were statistically significant differences between the left transverse sinus agenesis and the presence of the occipital sinus (p < 0.001), between the co-presence of the partial confluence type torcular and the occipital sinus (p = 0.040), and between the co-presence of the fenestrated straight sinus and the occipital sinus (p = 0.010). Although anatomical variations of dural venous sinuses are seen frequently, classification of venous sinuses helps surgeons in preoperative evaluation and management, and prevention of possible complications. In this study, we think that a comprehensive evaluation and classification of dural venous sinuses is a significant contribution to the literature.

Mississippi National River and Recreation Area : confluence area shuttle plan

DOT National Transportation Integrated Search

2015-02-28

This report described a proposed passenger shuttle for the Confluence Area of the National Park Service's Mississippi National River and Recreation Area. The report includes a proposed route, description of the stops, proposed schedules, and cost est...

Channel mapping river miles 29–62 of the Colorado River in Grand Canyon National Park, Arizona, May 2009

USGS Publications Warehouse

Kaplinski, Matt; Hazel, Joseph E.; Grams, Paul E.; Kohl, Keith; Buscombe, Daniel D.; Tusso, Robert B.

2017-03-23

Bathymetric, topographic, and grain-size data were collected in May 2009 along a 33-mi reach of the Colorado River in Grand Canyon National Park, Arizona. The study reach is located from river miles 29 to 62 at the confluence of the Colorado and Little Colorado Rivers. Channel bathymetry was mapped using multibeam and singlebeam echosounders, subaerial topography was mapped using ground-based total-stations, and bed-sediment grain-size data were collected using an underwater digital microscope system. These data were combined to produce digital elevation models, spatially variable estimates of digital elevation model uncertainty, georeferenced grain-size data, and bed-sediment distribution maps. This project is a component of a larger effort to monitor the status and trends of sand storage along the Colorado River in Grand Canyon National Park. This report documents the survey methods and post-processing procedures, digital elevation model production and uncertainty assessment, and procedures for bed-sediment classification, and presents the datasets resulting from this study.

An experiment to control nonnative fish in the Colorado River, Grand Canyon, Arizona

USGS Publications Warehouse

Coggins,, Lewis G.; Yard, Michael D.

2011-01-01

The humpback chub (Gila cypha) is an endangered native fish found only in the Colorado River Basin. In Grand Canyon, most humpback chub are found in the Little Colorado River and its confluence with the Colorado River. For decades, however, nonnative rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta), which prey on and compete with native fish, have dominated the Grand Canyon fish community. Between 2003 and 2006, scientists with the U.S. Geological Survey and Arizona Game and Fish Department experimentally removed 23,266 nonnative fish from a 9.4-mile-long reach of the Colorado River near where it joins the Little Colorado River. During the experiment, rainbow trout were reduced by as much as 90% and native fish abundance apparently increased in the reach. Concurrent environmental changes and a decrease in rainbow trout throughout the river make it difficult to determine if the apparent increase in native fish was the result of the experiment.

Geomorphic adjustment to hydrologic modifications along a meandering river: Implications for surface flooding on a floodplain

NASA Astrophysics Data System (ADS)

Edwards, Brandon L.; Keim, Richard F.; Johnson, Erin L.; Hupp, Cliff R.; Marre, Saraline; King, Sammy L.

2016-09-01

Responses of large regulated rivers to contemporary changes in base level are not well understood. We used field measurements and historical analysis of air photos and topographic maps to identify geomorphic trends of the lower White River, Arkansas, USA, in the 70 years following base-level lowering at its confluence with the Mississippi River and concurrent with flood control by dams. Incision was identified below a knickpoint area upstream of St. Charles, AR, and increases over the lowermost 90 km of the study site to 2 m near the confluence with the Mississippi River. Mean bankfull width increased by 30 m (21%) from 1930 to 2010. Bank widening appears to be the result of flow regulation above the incision knickpoint and concomitant with incision below the knickpoint. Hydraulic modeling indicated that geomorphic adjustments likely reduced flooding by 58% during frequent floods in the incised, lowermost floodplain affected by backwater flooding from the Mississippi River and by 22% above the knickpoint area. Dominance of backwater flooding in the incised reach indicates that incision is more important than flood control on the lower White River in altering flooding and also suggests that the Mississippi River may be the dominant control in shaping the lower floodplain. Overall, results highlight the complex geomorphic adjustment in large river-floodplain systems in response to anthropogenic modifications and their implications, including reduced river-floodplain connectivity.

Effects of Titanium Surface Microtopography and Simvastatin on Growth and Osteogenic Differentiation of Human Mesenchymal Stem Cells in Estrogen-Deprived Cell Culture.

PubMed

Arpornmaeklong, Premjit; Pripatnanont, Prisana; Chookiatsiri, Chonticha; Tangtrakulwanich, Boonsin

This study aimed to investigate the effects of titanium surface topography and simvastatin on growth and osteogenic differentiation of human bone marrow stromal cells (hBMSCs) in estrogen-deprived (ED) cell culture. Human BMSCs were seeded on cell culture plates, smooth-surface titanium (Ti) disks, and sandblasted with large grits and acid etched (SLA)-surface Ti disks; and subsequently cultured in regular (fetal bovine serum [FBS]), ED, and ED-with 100 nM simvastatin (ED-SIM) culture media for 14 to 21 days. Live/dead cell staining, scanning electron microscope examination, and cell viability assay were performed to determine cell attachment, morphology, and growth. Expression levels of osteoblast-associated genes, Runx2 and bone sialoprotein and levels of alkaline phosphatase (ALP) activity, calcium content, and osteocalcin in culture media were measured to determine osteoblastic differentiation. Expression levels of bone morphogenetic protein-2 (BMP-2) were investigated to examine stimulating effects of simvastatin (n = 4 to 5, mean ± SD). In vitro mineralization was verified by calcein staining. Human BMSCs exhibited different attachment and shapes on smooth and SLA titanium surfaces. Estrogen-deprived cell culture decreased cell attachment and growth, particularly on the SLA titanium surface, but cells were able to grow to reach confluence on day 21 in the ED-osteogenic (OS) culture medium. Promoting effects of the SLA titanium surface in ED-OS were significantly decreased. Simvastatin significantly increased osteogenic differentiation of human BMSCs on the SLA titanium surface in the ED-OS medium, and the promoting effects of simvastatin corresponded with the increasing of BMP-2 gene expression on the SLA titanium surface in ED-OS-SIM culture medium. The ED cell culture model provided a well-defined platform for investigating the effects of hormones and growth factors on cells and titanium surface interaction. Titanium, the SLA surface, and simvastatin synergistically promoted osteoblastic differentiation of hBMSCs in ED condition and might be useful to promote osteointegration in osteoporotic bone.

Multibody Parachute Flight Simulations for Planetary Entry Trajectories Using "Equilibrium Points"

NASA Technical Reports Server (NTRS)

Raiszadeh, Ben

2003-01-01

A method has been developed to reduce numerical stiffness and computer CPU requirements of high fidelity multibody flight simulations involving parachutes for planetary entry trajectories. Typical parachute entry configurations consist of entry bodies suspended from a parachute, connected by flexible lines. To accurately calculate line forces and moments, the simulations need to keep track of the point where the flexible lines meet (confluence point). In previous multibody parachute flight simulations, the confluence point has been modeled as a point mass. Using a point mass for the confluence point tends to make the simulation numerically stiff, because its mass is typically much less that than the main rigid body masses. One solution for stiff differential equations is to use a very small integration time step. However, this results in large computer CPU requirements. In the method described in the paper, the need for using a mass as the confluence point has been eliminated. Instead, the confluence point is modeled using an "equilibrium point". This point is calculated at every integration step as the point at which sum of all line forces is zero (static equilibrium). The use of this "equilibrium point" has the advantage of both reducing the numerical stiffness of the simulations, and eliminating the dynamical equations associated with vibration of a lumped mass on a high-tension string.

Using digital inline holographic microscopy and quantitative phase contrast imaging to assess viability of cultured mammalian cells

NASA Astrophysics Data System (ADS)

Missan, Sergey; Hrytsenko, Olga

2015-03-01

Digital inline holographic microscopy was used to record holograms of mammalian cells (HEK293, B16, and E0771) in culture. The holograms have been reconstructed using Octopus software (4Deep inwater imaging) and phase shift maps were unwrapped using the FFT-based phase unwrapping algorithm. The unwrapped phase shifts were used to determine the maximum phase shifts in individual cells. Addition of 0.5 mM H2O2 to cell media produced rapid rounding of cultured cells, followed by cell membrane rupture. The cell morphology changes and cell membrane ruptures were detected in real time and were apparent in the unwrapped phase shift images. The results indicate that quantitative phase contrast imaging produced by the digital inline holographic microscope can be used for the label-free real time automated determination of cell viability and confluence in mammalian cell cultures.

Substrate effects on endothelial cell adherence rates.

PubMed

Scott, W J; Mann, P

1990-01-01

Endothelial cell attachment to a synthetic substrate was studied using an in vitro model system. Attachment rate was defined as the number of tritium-labeled endothelial cells attached to a synthetic substrate after 30 minutes. The surface of the synthetic substrate was chemically modified with either laminin or fibronectin. Labeled endothelial cells attached more rapidly to synthetic substrate, chemically modified with biomolecules, as compared with the untreated substrate controls. Unlabeled endothelial cells were grown to confluency on a second set of modified and untreated substrates. The cells were removed with 1% Triton, and the rate of re-endothelialization with tritium-labeled endothelial cells was determined. The rate was 11-13 times that of the same cells on untreated substrate. These data confirm that biomolecules increase the attachment rate of endothelial cells to synthetic substrate, and also suggest that endothelial cells may secrete a Triton-insoluble product (Sigma, St. Louis, MO) into subendothelial matrix that increases re-endothelialization.

Heterologous desensitization of both phosphoinositide and Ca2+ signaling in SH-SY5Y neuroblastoma cells: a role for intracellular Ca2+ store depletion?

PubMed

Willars, G B; Nahorski, S R

1995-03-01

Measurement of the intracellular Ca2+ concentration ([Ca2+]i) in fura-2-loaded single cells of the human neuroblastoma line SH-SY5Y indicated coexpression of muscarinic and bradykinin receptors linked to activation of phosphoinositidase C (PIC). Both agonists elevated [Ca2+]i and inositol-1,4,5-trisphosphate [Ins(1,4,5)P3] levels in populations of adherent cells, although in cells used directly upon attainment of confluence the responses to carbachol were greater than those to bradykinin and displayed additional sustained components. This model system was used to examine heterologous interactions when a second PIC-linked agonist was added 100-300 sec after but in the continued presence of the first. Maximal (1 mM) carbachol concentrations abolished the elevation of [Ca2+]i produced by bradykinin but the muscarinic antagonist atropine (10 microM) restored the response, provided that extracellular Ca2+ was present throughout the experiment or was added before bradykinin. Carbachol also abolished bradykinin-mediated Ins(1,4,5)P3 elevation. In contrast, bradykinin did not influence [Ca2+]i or Ins(1,4,5)P3 responses to carbachol in the presence of extracellular Ca2+. In cells maintained at confluence for 2 weeks, the rapid peak elevations of [Ca2+]i and Ins(1,4,5)P3 levels induced by carbachol and bradykinin were approximately equivalent in magnitude. In these cells carbachol again abolished bradykinin-mediated elevation of [Ca2+]i but only attenuated, rather than abolished, the elevation of Ins(1,4,5)P3 levels. The [Ca2+]i and Ins(1,4,5)P3 responses to bradykinin were fully restored 100 sec after atropine only in the presence of extracellular Ca2+. Thus, depletion of an intracellular Ins(1,4,5)P3-sensitive Ca2+ store may underlie the ability of carbachol to produce not only heterologous desensitization of the [Ca2+]i elevation induced by bradykinin but also that of the Ins(1,4,5)P3 response. This suggests a feed-forward activation of PIC by Ca2+ released from Ins(1,4,5)P3-sensitive stores. Furthermore, studies in which Ins(1,4,5)P3-sensitive stores were depleted with thapsigargin and cells were challenged in the presence or absence of extracellular Ca2+ indicated that Ca2+, irrespective of its origin (intra- or extracellular), potentiated the Ins(1,4,5)P3 response to bradykinin alone. In cells maintained at confluence for 2 weeks, bradykinin was again unable to influence either [Ca2+]i or Ins(1,4,5)P3 responses to carbachol in the presence of Ca2+. This lack of heterologous desensitization may be due to the rapid, full, homologous desensitization of bradykinin receptors, compared with an incomplete homologous desensitization of muscarinic receptors.

In vitro analysis of human periodontal microvascular endothelial cells.

PubMed

Tsubokawa, Mizuki; Sato, Soh

2014-08-01

Endothelial cells (ECs) participate in key aspects of vascular biology, such as maintenance of capillary permeability, initiation of coagulation, and regulation of inflammation. According to previous reports, ECs have revealed highly specific characteristics depending on the organs and tissues. However, some reports have described the characteristics of the capillaries formed by human periodontal ECs. Therefore, the aim of the present study is to examine the functional characteristics of the periodontal microvascular ECs in vitro. Human periodontal ligament-endothelial cells (HPDL-ECs) and human gingiva-endothelial cells (HG-ECs) were isolated by immunoprecipitation with magnetic beads conjugated to a monoclonal anti-CD31 antibody. The isolated HPDL-ECs and HG-ECs were characterized to definitively demonstrate that these cell cultures represented pure ECs. Human umbilical-vein ECs and human dermal microvascular ECs were used for comparison. These cells were compared according to the proliferation potential, the formation of capillary-like tubes, the transendothelial electric resistance (TEER), and the expression of tight junction proteins. HPDL-ECs and HG-ECs with characteristic cobblestone monolayer morphology were obtained, as determined by light microscopy at confluence. Furthermore, the HPDL-ECs and HG-ECs expressed the EC markers platelet endothelial cell adhesion molecule-1 (also known as CD31), von Willebrand factor, and Ulex europaeus agglutinin 1, and the cells stained strongly positive for CD31 and CD309. In addition, the HPDL-ECs and HG-ECs were observed to form capillary-like tubes, and they demonstrated uptake of acetylated low-density lipoprotein. Functional analyses of the HPDL-ECs and HG-ECs showed that, compared to the control cells, tube formation persisted for only a brief period of time, and TEER was substantially reduced at confluence. Furthermore, the cells exhibited delocalization of zonula occludens-1 and occludin at cell-cell contact sites. The present results provide new evidence that HPDL-ECs and HG-ECs have characteristics of fenestrated capillaries. Therefore, capillaries in human periodontal tissues have functional characteristics of fenestrated capillaries, which might be related to the onset and the progression of systemic diseases and inflammation.

Keratinocyte Spray Technology for the Improved Healing of Cutaneous Sulfur Mustard Injuries

DTIC Science & Technology

2009-07-01

evaluations using cells harvested at lower degrees of confluence. REFERENCES Arroyo CM, Schafer RJ, Kurt EM, Broomfield CA, Carmichael AJ...Broomfield CA, Carmichael AJ. (1999) Response of normal human keratinocytes to sulfur mustard (HD): cytokine release using a non-enzymatic...Westchester Hall SUNY, NY 11794-8702 Voice: (n/a) Kertinocyte Spray, Jan30_2009Patient: USex: N/AAge: ?DOB: 30- Jan -09Biopsy Taken: 02-Feb-09Biopsy Received

Breast Cancer Stimulation of Osteolysis

DTIC Science & Technology

2000-09-01

essential medium supplemented with 10% fetal bovine serum (Hyclone, Logan, UT) and antibiotic/antimycotic solution (Sigma, St. Louis, MO) in 5% CO2 at...grown to confluence in alpha-modified minimum essential medium (Sigma) supplemented with 10% fetal bovine serum (Hyclone) at 37°C, 5% CO 2. ST2 cells...phenol red -free minimum essential medium supplemented with 10% FBS, and 50 ng/ml ascorbic acid. 1, 25-dihydroxyvitamin D3 and dexamethasone were

Understanding restoration of Oregon's Wood River through multi-modal hydrogeomorphic monitoring

NASA Astrophysics Data System (ADS)

Dearman, T.; Hughes, M. L.

2017-12-01

Channelized reaches of the lower Wood River in the Upper Klamath Basin of Oregon have undergone extensive restoration since the late 1990's, when the Bureau of Land Management began managing for the benefit of redband trout and other native-endemic species. Restoration included reconstruction of a floodplain and channel meanders, narrowing and deepening of channel, and excavation and reoccupation of fluvio-deltaic channels connecting the river to the Upper Klamath-Agency Lake system. The goals of this study were to extend the restoration monitoring record and evaluate post-restoration performance in light of this record. Monitoring included channel-bathymetry mapping, measurements of sediment transport (bedload), and measurement of discharge at points throughout the project reach under differing stage conditions. Results indicate two distinct domains of channel response to restoration: (1) an upstream domain marked by aggradation in the early and incision in the late post-restoration periods, and (2) a downstream domain marked by the inverse responses of degradation in the early and aggradation in the late post-restoration periods. These domains are separated by the confluence of an artificial channel maintained for boating access. Flow and sediment-transport continuity are interrupted at this confluence. At high stage (winter/spring) impoundment from the lake stalls flow, inducing sediment deposition. Stage falls as lake level recedes in the summer and stream power is restored, thereby releasing the sediment trapped at high stage. Aggradation in the downstream domain coupled with excavation of a birdfoot distributary in 2010 combined to initiate an avulsion from one distributary to another during the 2015 flow recession. With the exception of this recent avulsion, monitoring data suggest the channel system is approaching a dynamic equilibrium and behaving consistently with the rate law in geomorphology. This study provides the first known synthesis of long-term geomorphic monitoring of a freshwater fluvio-deltaic channel restoration in the Pacific Northwest.

ErbB2 and EGFR are downmodulated during the differentiation of 3T3-L1 preadipocytes.

PubMed

Pagano, Eleonora; Calvo, Juan Carlos

2003-10-15

The expression of receptors belonging to the epidermal growth factor receptor subfamily has been largely studied these last years in epithelial cells mainly as involved in cell proliferation and malignant progression. Although much work has focused on the role of these growth factor receptors in the differentiation of a variety of tissues, there is little information in regards to normal stromal cells. We investigated erbB2 expression in the murine fibroblast cell line Swiss 3T3L1, which naturally or hormonally induced undergoes adipocyte differentiation. We found that the Swiss 3T3-L1 fibroblasts express erbB2, in addition to EGFR, and in a quantity comparable to or even greater than the breast cancer cell line T47D. Proliferating cells increased erbB2 and EGFR levels when reaching confluence up to 4- and 10-fold, respectively. This expression showed a significant decrease when growth-arrested cells were stimulated to differentiate with dexamethasone and isobutyl-methylxanthine. Differentiated cells presented a decreased expression of both erbB2 and EGFR regardless of whether the cells were hormonally or spontaneously differentiated. EGF stimulation of serum-starved cells increased erbB2 tyrosine phosphorylation and retarded erbB2 migration in SDS-PAGE, suggesting receptor association and activation. Heregulin-alpha1 and -beta1, two EGF related factors, had no effect on erbB2 or EGFR phosphorylation. Although 3T3-L1 cells expressed heregulin, its specific receptors, erbB3 and erbB4, were not found. This is the first time in which erbB2 is reported to be expressed in an adipocytic cell line which does not depend on non EGF family growth factors (thyroid hormone, growth hormone, etc.) to accomplish adipose differentiation. Since erbB2 and EGFR expression were downmodulated as differentiation progressed it is conceivable that a mechanism of switching from a mitogenic to a differentiating signaling pathway may be involved, through regulation of the expression of these growth factor receptors. Copyright 2003 Wiley-Liss, Inc.

ECOLOGICAL STRUCTURE AMONG MIGRANT AND RESIDENT SEABIRDS OF THE SCOTIA-WEDDELL CONFLUENCE REGION

EPA Science Inventory

Report quantitatively assesses seasonal changes in community structure and habitat selection among seabirds in the Scotia-Weddell Confluence region, Antarctica. iscussed are biological and physical factors underlying the patterns. ata were derived from strip-transects on closely-...

76 FR 29656 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-23

... with +632 Unincorporated Areas of Kentucky River). Cow Creek to Estill County. approximately 0.7 mile... of Clearcreek Road. Cow Creek (backwater effects from From the confluence with +632 Unincorporated... Cow Creek Road. Crooked Creek (backwater effects from From the confluence with +631 Unincorporated...

Effect of low-level laser irradiation and epidermal growth factor on adult human adipose-derived stem cells.

PubMed

Mvula, B; Moore, T J; Abrahamse, H

2010-01-01

The study investigated the effects of low-level laser radiation and epidermal growth factor (EGF) on adult adipose-derived stem cells (ADSCs) isolated from human adipose tissue. Isolated cells were cultured to semi-confluence, and the monolayers of ADSCs were exposed to low-level laser at 5 J/cm(2) using 636 nm diode laser. Cell viability and proliferation were monitored using adenosine triphosphate (ATP) luminescence and optical density at 0 h, 24 h and 48 h after irradiation. Application of low-level laser irradiation at 5 J/cm(2) on human ADSCs cultured with EGF increased the viability and proliferation of these cells. The results indicate that low-level laser irradiation in combination with EGF enhances the proliferation and maintenance of ADSCs in vitro.

Regulation of expression of collagenase-3 in normal, differentiating rat osteoblasts

NASA Technical Reports Server (NTRS)

Winchester, S. K.; Bloch, S. R.; Fiacco, G. J.; Partridge, N. C.

1999-01-01

We investigated the regulation of collagenase-3 expression in normal, differentiating rat osteoblasts. Fetal rat calvarial cell cultures showed an increase in alkaline phosphatase activity reaching maximal levels between 7-14 days post-confluence, then declining with the onset of mineralization. Collagenase-3 mRNA was just detectable after proliferation ceased at day 7, increased up to day 21, and declined at later ages. Postconfluent cells maintained in non-mineralizing medium expressed collagenase-3 but did not show the developmental increase exhibited by cells switched to mineralization medium. Cells maintained in non-mineralizing medium continued to proliferate; cells in mineralization medium ceased proliferation. In addition, collagenase-3 mRNA was not detected in subcultured cells allowed to remineralize. These results suggest that enhanced accumulation of collagenase-3 mRNA is triggered by cessation of proliferation or acquisition of a mineralized extracellular matrix and that other factors may also be required. After initiation of basal expression, parathyroid hormone (PTH) caused a dose-dependent increase in collagenase-3 mRNA. Both the cyclic adenosine monophosphate (cAMP) analogue, 8-bromo-cAMP (8-Br-cAMP), and the protein kinase C (PKC) activator, phorbol myristate acetate, increased collagenase-3 expression, while the calcium ionophore, ionomycin, did not, suggesting that PTH was acting through the protein kinase A (PKA) and PKC pathways. Inhibition of protein synthesis with cycloheximide caused an increase in basal collagenase-3 expression but blocked the effect of PTH, suggesting that an inhibitory factor prevents basal expression while an inductive factor is involved with PTH action. In summary, collagenase-3 is expressed in mineralized osteoblasts and cessation of proliferation and initiation of mineralization are triggers for collagenase-3 expression. PTH also stimulates expression of the enzyme through both PKA and PKC pathways in the mineralizing osteoblast. Copyright 1999 Wiley-Liss, Inc.

Borneo vortex and mesoscale convective rainfall

NASA Astrophysics Data System (ADS)

Koseki, S.; Koh, T.-Y.; Teo, C.-K.

2014-05-01

We have investigated how the Borneo vortex develops over the equatorial South China Sea under cold surge conditions in December during the Asian winter monsoon. Composite analysis using reanalysis and satellite data sets has revealed that absolute vorticity and water vapour are transported by strong cold surges from upstream of the South China Sea to around the Equator. Rainfall is correspondingly enhanced over the equatorial South China Sea. A semi-idealized experiment reproduced the Borneo vortex over the equatorial South China Sea during a "perpetual" cold surge. The Borneo vortex is manifested as a meso-α cyclone with a comma-shaped rainband in the northeast sector of the cyclone. Vorticity budget analysis showed that the growth/maintenance of the meso-α cyclone was achieved mainly by the vortex stretching. This vortex stretching is due to the upward motion forced by the latent heat release around the cyclone centre. The comma-shaped rainband consists of clusters of meso-β-scale rainfall cells. The intense rainfall in the comma head (comma tail) is generated by the confluence of the warmer and wetter cyclonic easterly flow (cyclonic southeasterly flow) and the cooler and drier northeasterly surge in the northwestern (northeastern) sector of the cyclone. Intense upward motion and heavy rainfall resulted due to the low-level convergence and the favourable thermodynamic profile at the confluence zone. In particular, the convergence in the northwestern sector is responsible for maintenance of the meso-α cyclone system. At both meso-α and meso-β scales, the convergence is ultimately caused by the deviatoric strain in the confluence wind pattern but is significantly self-enhanced by the nonlinear dynamics.

Factors controlling the establishment of Fremont cottonwood seedlings on the Upper Green River, USA

USGS Publications Warehouse

Cooper, David J.; Merritt, David M.; Andersen, Douglas C.; Chimner, Rodney A.

1999-01-01

Declines in cottonwood (Populus spp.) recruitment along alluvial reaches of large rivers in arid regions of the western United States have been attributed to modified flow regimes, lack of suitable substrate, insufficient seed rain, and increased interspecific competition. We evaluated whether and how these factors were operating during 1993–1996 to influence demographics of Fremont cottonwood (P. deltoides Marshall subsp. wislizenii (Watson) Eckenwalder) along reaches of the Green and Yampa Rivers near their confluence in northwestern Colorado. We examined seedling establishment, defined as survival through three growing seasons, at three alluvial reaches that differed primarily in the level of flow regulation: a site on the unregulated Yampa, an upper Green River site regulated by Flaming Gorge Dam, and a lower Green River site below the Green–Yampa confluence. Seed rain was abundant in all sites, and led to large numbers of germinants (first-year seedlings) appearing each year at all sites. The regulated flow in the upper Green River reach restricted germination to islands and cut banks that were later inundated or eroded; no seedlings survived there. Mortality at the lower Green River site was due largely to desiccation or substrate erosion; 23% of 1993 germinants survived their first growing season, but at most 2% survived through their second. At the Yampa River site, germinants appeared on vegetated and unvegetated surfaces up to 2.5 m above base flow stage, but survived to autumn only on bare surfaces at least 1.25 m above base flow stage, and where at least 10 of the upper 40 cm of the alluvium was fine-textured. Our studies of rooting depths and the stable isotopic composition of xylem water showed that seedlings in the most favorable locations for establishment at the Yampa site do not become phreatophytic until their third or fourth growing season. Further, the results of experimental field studies examining effects of shade and competition supported the hypothesis that insufficient soil moisture, possibly in combination with insufficient light, restricts establishment to unvegetated sites. Collectively, the demographic and experimental studies suggest that, in arid regions, soil water availability is at least as important as light level in limiting establishment of Fremont cottonwood seedlings. We hypothesize that in cases where arid land rivers experience large spring stage changes, recruitment is further constrained within bare areas to those sites that contain sufficient fine-textured alluvium, saturated during the spring flood, to provide the flood-derived soil moisture normally necessary for late-summer seedling survival.

The density and biomass of mesozooplankton and ichthyoplankton in the Negro and the Amazon Rivers during the rainy season: the ecological importance of the confluence boundary

PubMed Central

Rimachi, Elvis V.; Santos-Silva, Edinaldo N.; Calixto, Laura S.F.; Leite, Rosseval G.; Khen, Adi; Yamane, Tetsuo; Mazeroll, Anthony I.; Inuma, Jomber C.; Utumi, Erika Y.K.; Tanaka, Akira

2017-01-01

The boundary zone between two different hydrological regimes is often a biologically enriched environment with distinct planktonic communities. In the center of the Amazon River basin, muddy white water of the Amazon River meets with black water of the Negro River, creating a conspicuous visible boundary spanning over 10 km along the Amazon River. Here, we tested the hypothesis that the confluence boundary between the white and black water rivers concentrates prey and is used as a feeding habitat for consumers by investigating the density, biomass and distribution of mesozooplankton and ichthyoplankton communities across the two rivers during the rainy season. Our results show that mean mesozooplankton density (2,730 inds. m−3) and biomass (4.8 mg m−3) were higher in the black-water river compared to the white-water river (959 inds. m−3; 2.4 mg m−3); however an exceptionally high mesozooplankton density was not observed in the confluence boundary. Nonetheless we found the highest density of ichthyoplankton in the confluence boundary (9.7 inds. m−3), being up to 9-fold higher than in adjacent rivers. The confluence between white and black waters is sandwiched by both environments with low (white water) and high (black water) zooplankton concentrations and by both environments with low (white water) and high (black water) predation pressures for fish larvae, and may function as a boundary layer that offers benefits of both high prey concentrations and low predation risk. This forms a plausible explanation for the high density of ichthyoplankton in the confluence zone of black and white water rivers. PMID:28507821

The density and biomass of mesozooplankton and ichthyoplankton in the Negro and the Amazon Rivers during the rainy season: the ecological importance of the confluence boundary.

PubMed

Nakajima, Ryota; Rimachi, Elvis V; Santos-Silva, Edinaldo N; Calixto, Laura S F; Leite, Rosseval G; Khen, Adi; Yamane, Tetsuo; Mazeroll, Anthony I; Inuma, Jomber C; Utumi, Erika Y K; Tanaka, Akira

2017-01-01

The boundary zone between two different hydrological regimes is often a biologically enriched environment with distinct planktonic communities. In the center of the Amazon River basin, muddy white water of the Amazon River meets with black water of the Negro River, creating a conspicuous visible boundary spanning over 10 km along the Amazon River. Here, we tested the hypothesis that the confluence boundary between the white and black water rivers concentrates prey and is used as a feeding habitat for consumers by investigating the density, biomass and distribution of mesozooplankton and ichthyoplankton communities across the two rivers during the rainy season. Our results show that mean mesozooplankton density (2,730 inds. m -3 ) and biomass (4.8 mg m -3 ) were higher in the black-water river compared to the white-water river (959 inds. m -3 ; 2.4 mg m -3 ); however an exceptionally high mesozooplankton density was not observed in the confluence boundary. Nonetheless we found the highest density of ichthyoplankton in the confluence boundary (9.7 inds. m -3 ), being up to 9-fold higher than in adjacent rivers. The confluence between white and black waters is sandwiched by both environments with low (white water) and high (black water) zooplankton concentrations and by both environments with low (white water) and high (black water) predation pressures for fish larvae, and may function as a boundary layer that offers benefits of both high prey concentrations and low predation risk. This forms a plausible explanation for the high density of ichthyoplankton in the confluence zone of black and white water rivers.

Evaluation of Flow Paths and Confluences for Saltwater Intrusion and Its Influence on Fish Species Diversity in a Deltaic River Network

NASA Astrophysics Data System (ADS)

Shao, X.; Cui, B.; Zhang, Z.; Fang, Y.; Jawitz, J. W.

2016-12-01

Freshwater in a delta is often at risk of saltwater intrusion, which has been a serious issue in estuarine deltas all over the world. Salinity gradients and hydrologic connectivity in the deltas can be disturbed by saltwater intrusion, which can fluctuate frequently and locally in time and space to affect biotic processes and then to affect the distribution patterns of the riverine fishes throughout the river network. Therefore, identifying the major flow paths or locations at risk of saltwater intrusion in estuarine ecosystems is necessary for saltwater intrusion mitigation and fish species diversity conservation. In this study, we use the betweenness centrality (BC) as the weighted attribute of the river network to identify the critical confluences and detect the preferential flow paths for saltwater intrusion through the least-cost-path algorithm from graph theory approach. Moreover, we analyse the responses of the salinity and fish species diversity to the BC values of confluences calculated in the river network. Our results show that the most likely location of saltwater intrusion is not a simple gradient change from sea to land, but closely dependent on the river segments' characteristics. In addition, a significant positive correlation between the salinity and the BC values of confluences is determined in the Pearl River Delta. Changes in the BC values of confluences can produce significant variation in the fish species diversity. Therefore, the dynamics of saltwater intrusion are a growing consideration for understanding the patterns and subsequent processes driving fish community structure. Freshwater can be diverted into these major flow paths and critical confluences to improve river network management and conservation of fish species diversity under saltwater intrusion.

Echocolor Doppler morpho-functional study of the jugulo-subclavian confluence in chronic cerebro-spinal venous insufficiency and multiple sclerosis patients.

PubMed

Mandolesi, Sandro; d'Alessandro, Aldo; Desogus, Antonello Ignazio; Ciccone, Marco Matteo; Zito, Annapaola; Stammegna, Immacolata; Niglio, Tarcisio; Orsini, Augusto; Mandolesi, Dimitri; d'Alessandro, Alessandro; Revelli, Luca

2017-01-01

The aim of this work is to measure the mean diameter of the confluence jugulo- subclavian, the impact of different types of jugular confluences and the correlation between the types of confluences and the Valsalva maneuver (jugular reflux) in subjects with Chronic Cerebro-Spinal Venous Insufficiency (CCSVI) and Multiple Sclerosis. We investigated by Echo-Color-Doppler (ECD) 103 subjects (67 F 36M) of mean age 45 ± 12 years (a minimum of 22 to a maximum of 79 years, with a median of 44 and a modal value 42 years), mean EDSS of 4.7 and average disease duration of 12 years. The 103 right jugular veins investigated had an average diameter of 8.4 ± 2.4 mm (minimum 4.0, maximum 14.9 mm; median 7.9; modal value 7.6 mm). Three form types were found: 56 cylindrical, 29 conical and 18 funnel. Valsalva maneuver was positive in 30 patients. The 103 left jugular investigated had an average diameter of 8.9 ± 2.4 mm (minimum 2.8, maximum 14.4 mm; median of 8.8; modal value 8.7 mm). The form types were found: 42 cylindrical, 45 conical and 16 funnel. Valsalva maneuver was positive in 30 patients. The mean diameter of the jugular veins was 8.7 mm. Internal jugular veins with cylindrical morphology have a diameter smaller than other forms; this difference is statistically significant. The different morphology of the jugular vein confluence does not increase the possibility of a reflux because the positive Valsalva maneuvers are not statistically significant when compared to the various types. CCSVI, EchoColorDoppler Map, Jugulo-Subclavian Confluence Diameter.

78 FR 36099 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-17

... Communities affected elevation above ground [caret]Elevation in meters (MSL) Modified Maricopa County, Arizona... Unincorporated Areas of upstream of the Camp Maricopa County. Creek Tributary A confluence. Approximately 400... miles +2857 upstream of the Camp Creek Tributary C confluence. Camp Creek Tributary C2 Approximately 400...

Marine Mammal Habitat in Ecuador: Seasonal Abundance and Environmental Distribution

DTIC Science & Technology

2010-06-01

derived macronutrients ) is enhanced by iron inputs derived from the island platform. The confluence of the Equatorial Undercurrent and Peru Current...is initiated by the subsurface derived macronutrients ) is enhanced by iron inputs derived from the island platform. The confluence of the Equatorial

75 FR 19328 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-14

... Lexington Road East. Paint Lick Creek (Backwater effects From the confluence None +573 Unincorporated Areas...). with the Kentucky Estill County. River to approximately 769 feet downstream of Roberts Road. Billey... Creek Road. Buck Creek Tributary 1 (Backwater From the confluence None +635 Unincorporated Areas of...

Family Characteristics and Intellectual Growth: An Examination by Race.

ERIC Educational Resources Information Center

Barnes, Jonathan

1979-01-01

The confluence model for explaining the influence of birth order on intelligence was applied to a sample of 56 Black and 52 White students. The confluence model and parental occupation were deemed insufficient in accounting for racial differences in intelligence scores. (Author/JKS)

Growth of cultured corneal endothelial cells onto a vitreous carbon matrix.

PubMed

Wickham, M G; Cleveland, P H; Binder, P S; Akers, P H

1983-01-01

Fourth passage cells of a rabbit corneal endothelial line were grown for 1 week in flasks containing pieces of a reticulated vitreous carbon matrix. The rate of cell growth in flasks containing the matrix was consistent with that in control flasks. Small fragments of the vitreous carbon material lying on the flask floor were covered by the monolayers as the cells grew to confluency. Vertical growth of cells onto larger pieces of the matrix proceeded in a staged fashion with maximum cell density on pieces of the matrix closest to the floor of the flask. As defined by scanning electron microscopy, cell growth occurred to a level at least 600 microns above the floor of the flask and the confluent monolayer. This novel culture procedure should be a model situation for study of many different aspects of the in vitro capabilities of corneal endothelial cells.

75 FR 64165 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-10-19

... River. Dog Creek (backwater effects from Nolin From the county boundary +560 Unincorporated Areas of Lake). to approximately 0.6 Edmonson County. mile upstream of the confluence with Dog Creek Tributary 1... County. approximately 1.1 miles upstream of the confluence with the Green River. Wolf Creek (backwater...

77 FR 6976 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-10

... Datum. + North American Vertical Datum. Depth in feet above ground. [caret] Mean Sea Level, rounded to... Unincorporated Areas of approximately 0.5 mile Taney County. upstream of the White River confluence. Big Shoals... confluence. * National Geodetic Vertical Datum. + North American Vertical Datum. Depth in feet above ground...

76 FR 19007 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-06

... Birch Creek Road. Black Mingo Creek At the upstream side of None +14 Unincorporated Areas of County Line... Road. Boggy Swamp A At the Black Mingo None +25 Unincorporated Areas of Creek confluence. Williamsburg... mile None +35 Unincorporated Areas of upstream of the Black Williamsburg County. River confluence...

78 FR 9831 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2013-02-12

... Drive. Wake County. Approximately 850 feet +345 upstream of Keighley Forest Drive. Richland Creek Approximately 850 feet +301 Town of Wake Forest. upstream of the confluence with Richland Creek Tributary 2... Approximately 750 feet +301 Town of Wake Forest. upstream of the confluence with Richland Creek. Approximately 0...

75 FR 29290 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-25

... the intersection of Lodge None * 831 Road and Main Street/State Highway 112. National Geodetic... None +976 upstream of Mt. Vernon Road. Calloway Creek (Backwater effects From the confluence None +590.... approximately 635 feet upstream of Doylesville Road. Dreaming Creek Tributary At the confluence with None +837...

75 FR 29268 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-25

... the Owen County Courthouse, 100 North Thomas Street, Owenton, KY 40359. Trigg County, Kentucky, and... County. Route 116. Approximately 2.5 miles +839 +838 upstream of the confluence with Jenkins Branch... County. Approximately 0.9 mile None +944 upstream of Missouri Route DD. Jenkins Branch At the confluence...

Base flow (1966-2009) and streamflow gain and loss (2010) of the Brazos River from the New Mexico-Texas State line to Waco, Texas

USGS Publications Warehouse

Baldys, Stanley; Schalla, Frank E.

2012-01-01

Streamflow was measured at 66 sites from June 6–9, 2010, and at 68 sites from October 16–19, 2010, to identify reaches in the upper Brazos River Basin that were gaining or losing streamflow. Gaining reaches were identified in each of the five subbasins. The gaining reach in the Salt Fork Brazos River Basin began at USGS streamflow-gaging station 08080940 Salt Fork Brazos River at State Highway 208 near Clairemont, Tex. (site SF–6), upstream from where Duck Creek flows into the Salt Fork Brazos River and continued downstream past USGS streamflow-gaging station 08082000 Salt Fork Brazos River near Aspermont, Tex. (site SF–9), to the outlet of the basin. In the Double Mountain Fork Brazos River Basin, a gaining reach from near Post, Tex., downstream to the outlet of the basin was identified. Two gaining reaches were identified in the Clear Fork Brazos River Basin—one from near Roby, Tex., downstream to near Noodle, Tex., and second from Hawley, Tex., downstream to Nugent, Tex. Most of the North Bosque River was characterized as gaining streamflow. Streamflow gains were identified in the main stem of the Brazos River from where the Brazos River main stem forms at the confluence of the Salt Fork Brazos River and Double Mountain Fork Brazos River near Knox City, Tex., downstream to near Seymour, Tex.

Managing injuries of hepatic duct confluence variants after major hepatobiliary surgery: An algorithmic approach

PubMed Central

Fragulidis, Georgios; Marinis, Athanasios; Polydorou, Andreas; Konstantinidis, Christos; Anastasopoulos, Georgios; Contis, John; Voros, Dionysios; Smyrniotis, Vassilios

2008-01-01

AIM:To investigate injuries of anatomy variants of hepatic duct confluence during hepatobiliary surgery and their impact on morbidity and mortality of these procedures. An algorithmic approach for the management of these injuries is proposed. METHODS: During a 6-year period 234 patients who had undergone major hepatobiliary surgery were retrospectively reviewed in order to study postoperative bile leakage. Diagnostic workup included endoscopic and magnetic retrograde cholangiopancreatography (E/MRCP), scintigraphy and fistulography. RESULTS: Thirty (12.8%) patients who developed postoperative bile leaks were identified. Endoscopic stenting and percutaneous drainage were successful in 23 patients with bile leaks from the liver cut surface. In the rest seven patients with injuries of hepatic duct confluence, biliary variations were recognized and a stepwise therapeutic approach was considered. Conservative management was successful only in 2 patients. Volume of the liver remnant and functional liver reserve as well as local sepsis were used as criteria for either resection of the corresponding liver segment or construction of a biliary-enteric anastomosis. Two deaths occurred in this group of patients with hepatic duct confluence variants (mortality rate 28.5%). CONCLUSION: Management of major biliary fistulae that are disconnected from the mainstream of the biliary tree and related to injury of variants of the hepatic duct confluence is extremely challenging. These patients have a grave prognosis and an early surgical procedure has to be considered. PMID:18494057

Earth Observations taken by Expedition 38 crewmember

NASA Image and Video Library

2013-12-26

ISS038-E-021397 (24 Dec. 2013) --- The Caribbean country of Cuba is pictured in this nadir image, photographed by one of the Expedition 38 crew members aboard the International Space Station. (Note: North is at the top of the picture.) Cuba is an archipelago of islands in the northern Caribbean Sea at the confluence with the Gulf of Mexico and the Atlantic Ocean. In the southeast, the dark coastal area is home to the Sierra Maestra mountains. It is the highest mountain range on the island, with Pico Turquino reaching nearly 2000 meters. On the central southern coast of the island is the Sierra Del Escambray mountain range, including the 1160 meter-high Pico San Juan, Cuba's second highest peak.

Hydrodynamics and morphodynamics of a mobile bed confluence under equilibrium conditions for different values of the flow discharge ratio

NASA Astrophysics Data System (ADS)

Birjukova Canelas, Olga; Lage Ferreira, Rui Miguel; Heleno Cardoso, António

2017-04-01

Under steady water and sediment feeding in laboratory conditions, sediment fluxes tend to reach a steady state and bed morphology tends to equilibrium. This laboratory study states two objectives: i) to provide a detailed three-dimensional characterization of the flow field at a movable bed confluence and ii) to contribute to the characterization of the effect of the flow discharge ratio (Qr = Qt/Qm, Qt - tributary flow discharge and Qm - main channel flow discharge) on the flow field. While the junction angle between the main channel and the tributary as well as the sediment discharge ratio were kept constant, two scenarios corresponding to two different flow discharge ratios with dominant main channel flow discharge (Qm > Qt) were analyzed. Prior to the flow velocity measurements, both channels were fed with water and sand mixtures and tests were run until the equilibrium condition was reached, i.e. until the moment where the difference between the total supplied sand discharge and the total outgoing sand discharge was smaller than ± 5%. During the experiments, bed topography was systematically recorded. Then, the flow was stopped, the water was slowly drained and the bed was carefully fixed with a cement and vernix coat and allowed to dry to guarantee that it remained stable from there on, while the clear water (free of sand) discharges were fed again to the fixed bed converging channels. A 3D side looking Acoustic Doppler Velocimeter (ADV), installed on a remotely controlled and precisely positioned (± 0.1 mm) robotic arm, was used to measure the instantaneous velocities at a very dense mesh. Once the velocities measured and recorded, the water surface levels were surveyed. For the case with more dominant tributary flow (Qr = 0.17), as expected, the scour hole formed at the tributary mouth is much deeper and larger and extends towards the outer bank of the post-confluence channel, as well as upstream the junction corner. The avalanche slopes are, consequently, steeper. On the other hand, for the case with a weaker tributary flow (Qr = 0.08), no erosion is observed immediately upstream the junction. Moreover, the bank-attached sediment bar downstream the junction is considerably more developed in longitudinal and vertical directions. In both scenarios, a strong horizontal vortical structure was observed immediately downstream the junction. More detailed analysis of the origin and the characteristics of these and other flow patterns is still under investigation. Acknowledgements This research as partially supported by Portuguese and European funds, within programs COMPETE2020 and PORL-FEDER, through project PTDC/ECM-HID/6387/2014 granted by the National Foundation for Science and Technology (FCT).

Neutrophil-endothelial cell interactions on endothelial monolayers grown on micropore filters.

PubMed Central

Taylor, R F; Price, T H; Schwartz, S M; Dale, D C

1981-01-01

We have developed a technique for growing endothelial monolayers on micropore filters. These monolayers demonstrate confluence by phase and electron microscopy and provide a functional barrier to passage of radiolabeled albumin. Neutrophils readily penetrate the monolayer in response to chemotaxin, whereas there is little movement in the absence of chemotaxin. This system offers unique advantages over available chemotaxis assays and may have wider applications in the study of endothelial function. Images PMID:7007441

76 FR 40670 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2011-07-11

... Incorporated Areas Black Creek At the Contentnea Creek +69 +66 Town of Black Creek, confluence. Unincorporated Areas of Wilson County. Approximately 50 feet +92 +91 upstream of U.S. Route 117. Black Creek Tributary Approximately 1,500 +93 +92 Town of Lucama, feet upstream of the Unincorporated Areas Black Creek confluence. of...

76 FR 21695 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-18

... County. At the Betts Spring None +571 Branch and Bradford Creek confluence. Big Cove Creek Approximately.... Approximately 0.7 mile None +677 upstream of the Big Cove Creek confluence. Blue Spring Creek Approximately 400... Road. Approximately 450 feet None +748 upstream of Spragins Hollow Road Northwest. East Fork Pinhook...

27 CFR 9.105 - Cumberland Valley.

Code of Federal Regulations, 2010 CFR

2010-04-01

... Chesapeake & Ohio (C&O) Canal National Historical Park and the confluence of the Potomac River and... perimeter of the park on the northeastern bank of the Potomac River to the confluence of Antitam Creek and the Potomac River; (2) Then southeast of Limekiln Road which runs along the perimeter of the park from...

27 CFR 9.105 - Cumberland Valley.

Code of Federal Regulations, 2011 CFR

2011-04-01

... Chesapeake & Ohio (C&O) Canal National Historical Park and the confluence of the Potomac River and... perimeter of the park on the northeastern bank of the Potomac River to the confluence of Antitam Creek and the Potomac River; (2) Then southeast of Limekiln Road which runs along the perimeter of the park from...

27 CFR 9.105 - Cumberland Valley.

Code of Federal Regulations, 2012 CFR

2012-04-01

... Chesapeake & Ohio (C&O) Canal National Historical Park and the confluence of the Potomac River and... perimeter of the park on the northeastern bank of the Potomac River to the confluence of Antitam Creek and the Potomac River; (2) Then southeast of Limekiln Road which runs along the perimeter of the park from...

27 CFR 9.105 - Cumberland Valley.

Code of Federal Regulations, 2013 CFR

2013-04-01

... Chesapeake & Ohio (C&O) Canal National Historical Park and the confluence of the Potomac River and... perimeter of the park on the northeastern bank of the Potomac River to the confluence of Antitam Creek and the Potomac River; (2) Then southeast of Limekiln Road which runs along the perimeter of the park from...

27 CFR 9.105 - Cumberland Valley.

Code of Federal Regulations, 2014 CFR

2014-04-01

... Chesapeake & Ohio (C&O) Canal National Historical Park and the confluence of the Potomac River and... perimeter of the park on the northeastern bank of the Potomac River to the confluence of Antitam Creek and the Potomac River; (2) Then southeast of Limekiln Road which runs along the perimeter of the park from...

75 FR 19895 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-16

... +1529 Eastern Band of Cherokee Hanging Dog Creek. Indians, Unincorporated Areas of Cherokee County. Approximately 0.8 mile +1633 upstream of the confluence with Hanging Dog Creek. Big Witch Creek At the.... Indians. Approximately 1,050 feet +1904 upstream of Goose Creek Road. Hanging Dog Creek At the confluence...

75 FR 23642 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-04

... Flat River Approximately 375 feet None +683 City of Desloge, City downstream of the of Park Hills... confluence with Flat of St. Francois Creek. County. Approximately 2,000 None +767 feet upstream of City of... of the St. Francois County. confluence with the Flat River. Approximately 7,000 None +783 feet...

77 FR 21485 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2012-04-10

... Incorporated Areas Docket No.: FEMA-B-1085 Bee Creek Tributary B Approximately 1,700 feet + 286 City of College... Turkey Creek Tributary D. Unnamed Tributary to Bee Creek At the confluence with + 291 City of College Station. Tributary B. Bee Creek Tributary B. Approximately 613 feet + 293 upstream of the confluence with...

75 FR 29264 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-25

... Creek Tributary 1 At the downstream side None +341 Unincorporated Areas of of Keighley Forest Wake... Approximately 850 feet +302 +301 Town of Wake Forest. upstream of the confluence with Richland Creek Tributary 2... Approximately 750 feet None +301 Town of Wake Forest. upstream of the confluence with Richland Creek...

Dexamethasone effects on creatine kinase activity and insulin-like growth factor receptors in cultured muscle cells

NASA Technical Reports Server (NTRS)

Whitson, Peggy A.; Stuart, Charles A.; Huls, M. H.; Sams, Clarence F.; Cintron, Nitza M.

1989-01-01

The effect of dexamethasone on the activity of creatine kinase (CK) and the insulin-like growth factor I (IGF-I) binding were investigated using skeletal- and cardiac-muscle-derived cultured cell lines (mouse, C2C12; rat, L6 and H9c2). It was found that, in skeletal muscle cells, dexamethasone treatment during differentiation of skeletal-muscle cells caused dose-dependent increases in CK activity and increases in the degree of myotube formation, whereas cardiac cells (H9c2) exhibited very low CK activity during culture or dexamethasone treatment. Results for IGF-I binding were similar in all three cell lines. The IGF-I binding to dexamethasone-treated cells (50 nM for 24 hr on the day prior to confluence) resulted in an increased number of available binding sites, with no effect on the binding affinities.

Chromatin plasticity as a differentiation index during muscle differentiation of C2C12 myoblasts

DOE Office of Scientific and Technical Information (OSTI.GOV)

Watanabe, Tomonobu M.; World Premier Initiative, iFREC, Osaka University, Osaka 565-0871; Higuchi, Sayaka

Highlights: Black-Right-Pointing-Pointer Change in the epigenetic landscape during myogenesis was optically investigated. Black-Right-Pointing-Pointer Mobility of nuclear proteins was used to state the epigenetic status of the cell. Black-Right-Pointing-Pointer Mobility of nuclear proteins decreased as myogenesis progressed in C2C12. Black-Right-Pointing-Pointer Differentiation state diagram was developed using parameters obtained. -- Abstract: Skeletal muscle undergoes complicated differentiation steps that include cell-cycle arrest, cell fusion, and maturation, which are controlled through sequential expression of transcription factors. During muscle differentiation, remodeling of the epigenetic landscape is also known to take place on a large scale, determining cell fate. In an attempt to determine the extentmore » of epigenetic remodeling during muscle differentiation, we characterized the plasticity of the chromatin structure using C2C12 myoblasts. Differentiation of C2C12 cells was induced by lowering the serum concentration after they had reached full confluence, resulting in the formation of multi-nucleated myotubes. Upon induction of differentiation, the nucleus size decreased whereas the aspect ratio increased, indicating the presence of force on the nucleus during differentiation. Movement of the nucleus was also suppressed when differentiation was induced, indicating that the plasticity of chromatin changed upon differentiation. To evaluate the histone dynamics during differentiation, FRAP experiment was performed, which showed an increase in the immobile fraction of histone proteins when differentiation was induced. To further evaluate the change in the histone dynamics during differentiation, FCS was performed, which showed a decrease in histone mobility on differentiation. We here show that the plasticity of chromatin decreases upon differentiation, which takes place in a stepwise manner, and that it can be used as an index for the differentiation stage during myogenesis using the state diagram developed with the parameters obtained in this study.« less

Saturated and Unsaturated Fatty Acids Differently Modulate Colonic Goblet Cells In Vitro and in Rat Pups.

PubMed

Benoit, Bérengère; Bruno, Jérémie; Kayal, Fanny; Estienne, Monique; Debard, Cyrille; Ducroc, Robert; Plaisancié, Pascale

2015-08-01

High-fat diets induce intestinal barrier alterations and promote intestinal diseases. Little is known about the effects of long-chain fatty acids (LCFAs) on mucin 2 (MUC2) production by goblet cells, which are crucial for intestinal protection. We investigated the effects of LCFAs on the differentiation of colonic goblet cells, MUC2 expression, and colonic barrier function. Upon reaching confluence, human colonic mucus-secreting HT29-MTX cells were stimulated (21 d) with a saturated LCFA (palmitic or stearic acid), a monounsaturated LCFA (oleic acid), or a polyunsaturated LCFA (linoleic, γ-linolenic, α-linolenic, or eicosapentaenoic acid). In addition, rat pups underwent oral administration of oil (palm, rapeseed, or sunflower oil) or water (10 μL/g body weight, postnatal days 10-15). Subsequently, colon goblet cells were studied by Western blotting, reverse transcriptase-quantitative polymerase chain reaction, and immunohistochemistry and colonic transmucosal electrical resistance was measured by using Ussing chambers. In vitro, palmitic acid enhanced MUC2 production (140% of control) and hepatocyte nuclear factor 4α expression, whereas oleic, linoleic, γ-linolenic, α-linolenic, and eicosapentaenoic acids reduced MUC2 expression (at least -50% of control). All unsaturated LCFAs decreased the expression of human atonal homolog 1, a transcription factor controlling goblet cell differentiation (at least -31% vs. control). In vivo, rats fed palm oil had higher palmitic acid concentrations (3-fold) in their colonic contents and increased mucus granule surfaces in their goblet cells (>2-fold) than did all other groups. Palm oil also increased colonic transmucosal electrical resistance (245% of control), yet had no effect on occludin and zonula occludens-1 expression. In contrast, sunflower and rapeseed oils decreased goblet cell number when compared with control (at least -10%) and palm oil (at least -14%) groups. Palm oil in rat pups and palmitic acid in HT29-MTX cells increase the production of MUC2 and strengthen the intestinal barrier. In contrast, unsaturated LCFAs decrease MUC2 expression. These data should be taken into account in the context of preventive or therapeutic nutritional programs. © 2015 American Society for Nutrition.

A 3-D Navier-Stokes CFD study of turbojet/ramjet nozzle plume interactions at Mach 3.0 and comparison with data

NASA Technical Reports Server (NTRS)

Chang, Ing; Hunter, Louis G.

1995-01-01

Advanced airbreathing propulsion systems used in Mach 4-6 mission scenarios, usually consist of a single integrated turboramjet or as in this study, a turbojet housed in an upper bay with a separate ramjet housed in a lower bay. As the engines transition from turbojet to ramjet, there is an operational envelope where both engines operate simultaneously. One nozzle concept under consideration has a common nozzle, where the plumes from the turbojet and ramjet interact with one another as they expand to ambient conditions. In this paper, the two plumes interact at the end of a common 2-D cowl, when they both reach an approximate Mach 3.0 condition and then jointly expand to Mach 3.6 at the common nozzle exit plane. At this condition, the turbojet engine operated at a higher NPR than the ramjet, where the turbojet overpowers the ramjet plume, deflecting it approximately 12 degrees downward and in turn the turbojet plume is deflected 6 degrees upward. In the process, shocks were formed at the deflections and a shear layer formed at the confluence of the two jets. This particular case was experimentally tested and the data used to compare with the PARC3D code with k-kl two equation turbulence model. The 2-D and 3-D centerline CFD solutions are in good agreement, but as the CFD solutions approach the outer sidewall, a slight variance occurs. The outer wall boundary layers are thin and do not present much of an interaction, however, where the confluence interaction shocks interact with the thin boundary layer on the outer wall, strong vortices run down each shock causing substantial disturbances in the boundary layer. These disturbances amplify somewhat as they propagate downstream axially from the confluence point. The nozzle coefficient (CFG) is reduced 1/2 percent as a result of this sidewall interaction, from 0.9850 to 0.9807. This three-dimensional reduction is in better agreement with the experimental value of 0.9790.

Flood Study of Warren Brook in Alstead and Cold River in Alstead, Langdon, and Walpole, New Hampshire, 2005

USGS Publications Warehouse

Flynn, Robert H.

2006-01-01

This report presents water-surface elevations and profiles as determined using the U.S. Army Corps of Engineers (USACE) one-dimensional Hydrologic Engineering Center River Analysis System, also known as HEC-RAS. Steady flow water-surface profiles were developed for two stream reaches: the Cold River from its confluence with the Connecticut River in Walpole, through Alstead to the McDermott Bridge in Langdon, NH, and Warren Brook from its confluence with the Cold River to Warren Lake in Alstead, NH. Flood events of a magnitude, which are expected to be equaled or exceeded once on the average during any 10-, 50-, 100-, or 500-year period (recurrence interval), were modeled using HEC-RAS as these flood events are recognized as being significant for flood-plain management, determination of flood insurance rates, and design of structures such as bridges and culverts. These flood events are referred to as the 10-, 50-, 100-, and 500-year floods and have a 10-, 2-, 1-, and 0.2-percent chance, respectively, of being equaled or exceeded during any year. The recurrence intervals represent the long-term average between floods of a specific magnitude. The risk of experiencing rare floods at short intervals or within the same year increases when periods greater than one year are considered. The analyses in this study reflect the flooding potentials based on conditions existing in the communities of Walpole, Alstead and Langdon at the time of completion of this study.

76 FR 32896 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2011-06-07

... buildings. Comments on any aspect of the Flood Insurance Study and FIRM, other than the proposed BFEs, will... Creek confluence. At the downstream side +489 +491 of Oak Gate Lane. Long Branch (of Duck Creek) Bypass.. At the upstream side of +498 +490 City of Mesquite. the Long Branch (of Duck Creek) confluence...

Intellectual Development within Transracial Adoptive Families: Retesting the Confluence Model.

ERIC Educational Resources Information Center

Berbaum, Michael L.; Moreland, Richard L.

1985-01-01

Estimates confluence model of intellectual development for a within-family sample of 321 children from 101 transracial adoptive families. Mental ages of children and their parents and birth or adoption intervals were used in a nonlinear least-squares estimation procedure to obtain children's predicted mental ages. Results suggest efficiency of the…

Confluence of the Amazon and Topajos Rivers, Brazil, South America

NASA Image and Video Library

1991-08-11

This view shows the confluence of the Amazon and the Topajos Rivers at Santarem, Brazil (2.0S, 55.0W). The Am,azon flows from lower left to upper right of the photo. Below the river juncture of the Amazon and Tapajos, there is considerable deforestation activity along the Trans-Amazon Highway.

The current content of artificial radionuclides in the water of the Tobol-Irtysh river system (from the mouth of the Iset River to the confluence with the Ob River).

PubMed

Nikitin, Alexander I; Chumichev, Vladimir B; Valetova, Nailia K; Katrich, Ivan Yu; Kabanov, Alexander I; Dunaev, Gennady E; Shkuro, Valentina N; Rodin, Victor M; Mironenko, Alexander N; Kireeva, Elena V

2007-01-01

Data on content of (90)Sr, (137)Cs, (239,240)Pu and (3)H in water of the Tobol-Irtysh part of the Techa-Iset-Tobol-Irtysh-Ob river system (through which the "Mayak" PA radioactive wastes are transported) are presented and discussed. The data were received in 2004-2005 under the ISTC project on radioecological monitoring of the Tobol and Irtysh rivers. Monthly observations of (137)Cs, (90)Sr and (3)H content in water in the area of the Tobol and Irtysh confluence have been conducted starting from May 2004. To obtain information on the investigated river system as a whole, the radioecological survey of the Tobol and Irtysh rivers at the section from the mouth of the Iset River to the confluence with the Ob River was carried out in 2004. It is shown that the impact of "Mayak" PA waste transport by (90)Sr is distinctly traced as far as the area of the Irtysh and Ob confluence.

Borneo Vortex and Meso-scale Convective Rainfall

NASA Astrophysics Data System (ADS)

Koh, T. Y.; Koseki, S.; Teo, C. K.

2014-12-01

We have investigated how the Borneo vortex develops over the equatorial South China Sea under cold surge conditions in December during the Asian winter monsoon. Composite analysis using reanalysis and satellite datasets has revealed that absolute vorticity and water vapour are transported by strong cold surges from upstream of the South China Sea to around the equator. Rainfall is correspondingly enhanced over the equatorial South China Sea. A semi-idealized experiment reproduced the Borneo vortex over the equatorial South China Sea during a perpetual cold surge. The Borneo vortex is manifested as a meso-alpha cyclone with a comma-shaped rainband in the northeast sector of the cyclone. Vorticity budget analysis showed that the growth/maintenance of the meso-alpha cyclone was achieved mainly by the vortex stretching. This vortex stretching is due to the upward motion forced by the latent heat release around the cyclone centre. The comma-shaped rainband consists of clusters of meso-beta scale rainfall cells. The intense rainfall in the comma-head (comma-tail) is generated by the confluence of the warmer and wetter cyclonic easterly flow (cyclonic southeasterly flow) and the cooler and drier northeasterly surge in the northwestern (northeastern) sector of the cyclone. Intense upward motion and heavy rainfall resulted due to the low-level convergence and the favourable thermodynamic profile at the confluence zone. In particular, the convergence in the northwestern sector is responsible for maintenance of the meso-alpha cyclone system. At both meso-alpha and meso-beta scales, the convergence is ultimately caused by the deviatoric strain in the confluence wind pattern but is significantly self-enhanced by the nonlinear dynamics. Reference: Koseki, S., T.-Y. Koh and C.-K. Teo (2014), Atmospheric Chemistry and Physics, 14, 4539-4562, doi:10.5194/acp-14-4539-2014, 2014.

Enhancement of UV-induced nucleotide excision repair activity upon forskolin treatment is cell growth-dependent.

PubMed

Lee, Jeong-Min; Park, Jeong-Min; Kang, Tae-Hong

2016-10-01

Forskolin (FSK), an adenylyl cyclase activator, has recently been shown to enhance nucleotide excision repair (NER) upon UV exposure. However, our study revealed that this effect was detected in human skin epithelial ARPE19 cells only in growing cells, but not in non-cycling cells. When the cells were grown at low density (70% confluence), FSK was capable of stimulating cAMP responsive element binding (CREB) phosphorylation, a marker for FSK-stimulated PKA activation, and resulted in a significant increase of NER activity compared to control treatment. However, cells grown under 100% confluent conditions showed neither FSK-induced CREB phosphorylation nor the resulting NER enhancement. These findings indicate that cellular growth is critical for FSK-induced NER enhancement and suggest that cellular growth conditions should be considered as a variable while evaluating a reagent's pharmacotherapeutic efficacy. [BMB Reports 2016; 49(10): 566-571].

The Volume of Three-Dimensional Cultures of Cancer Cells InVitro Influences Transcriptional Profile Differences and Similarities with Monolayer Cultures and Xenografted Tumors.

PubMed

Boghaert, Erwin R; Lu, Xin; Hessler, Paul E; McGonigal, Thomas P; Oleksijew, Anatol; Mitten, Michael J; Foster-Duke, Kelly; Hickson, Jonathan A; Santo, Vitor E; Brito, Catarina; Uziel, Tamar; Vaidya, Kedar S

2017-09-01

Improving the congruity of preclinical models with cancer as it is manifested in humans is a potential way to mitigate the high attrition rate of new cancer therapies in the clinic. In this regard, three-dimensional (3D) tumor cultures in vitro have recently regained interest as they have been acclaimed to have higher similarity to tumors in vivo than to cells grown in monolayers (2D). To identify cancer functions that are active in 3D rather than in 2D cultures, we compared the transcriptional profiles (TPs) of two non-small cell lung carcinoma cell lines, NCI-H1650 and EBC-1 grown in both conditions to the TP of xenografted tumors. Because confluence, diameter or volume can hypothetically alter TPs, we made intra- and inter-culture comparisons using samples with defined dimensions. As projected by Ingenuity Pathway Analysis (IPA), a limited number of signal transduction pathways operational in vivo were better represented by 3D than by 2D cultures in vitro. Growth of 2D and 3D cultures as well as xenografts induced major changes in the TPs of these 3 modes of culturing. Alterations of transcriptional network activation that were predicted to evolve similarly during progression of 3D cultures and xenografts involved the following functions: hypoxia, proliferation, cell cycle progression, angiogenesis, cell adhesion, and interleukin activation. Direct comparison of TPs of 3D cultures and xenografts to monolayer cultures yielded up-regulation of networks involved in hypoxia, TGF and Wnt signaling as well as regulation of epithelial mesenchymal transition. Differences in TP of 2D and 3D cancer cell cultures are subject to progression of the cultures. The emulation of the predicted cell functions in vivo is therefore not only determined by the type of culture in vitro but also by the confluence or diameter of the 2D or 3D cultures, respectively. Consequently, the successful implementation of 3D models will require phenotypic characterization to verify the relevance of applying these models for drug development. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Temporal and spatial scales of geomorphic adjustments to reduced competency following flow regulation in bedload-dominated systems

NASA Astrophysics Data System (ADS)

Curtis, Katherine E.; Renshaw, Carl E.; Magilligan, Francis J.; Dade, William B.

2010-05-01

Because of the combined effects of reduced sediment transport capacity and competency following flow regulation, morphological changes are expected to occur in channels downstream from dams and, specifically, at tributary junctions where local inputs of water and sediment occur. Using a combination of historical aerial photographs, mainstem- and tributary-channel pebble counts, and HEC-RAS flow modeling for two watersheds in south-central VT, one unregulated and the other regulated since 1961, we document the time series of post-regulation channel narrowing and associated bar growth due to the influx of tributary sediment. Channel adjustments at regulated tributary junctions have been significant in ca. 50 years following impoundment, with channels downstream of the confluences narrowing over 15% after an initial ca. 20-year lag before the onset of accelerated narrowing. Moreover, flow modeling suggests that downstream of regulated confluences, the modern median grain size ( d50) along the channel bed is immobile. No significant channel narrowing has occurred either above or below unregulated tributary junctions or on the mainstem upstream of regulated confluences. However, greater channel sediment fining is observed upstream of regulated confluences than above unregulated confluences. Thus, the primary mode of mainstem channel adjustment differs up- and downstream of regulated tributaries. These confluence effects have occurred where the tributary drainage area is only 0.2 times that of the mainstem, well below the threshold ratio of 0.6 required for significant geomorphic effects at unregulated confluences, highlighting the geomorphic scale shift of dams. Lastly, we evaluate the downstream length required for a river to recover from the impacts of impoundment and demonstrate that even distal locations are impacted by flow regulation. Unlike the impacts of flow regulation in the western US where channel incision and bar erosion predominate following impoundment, we find that in situations where bed incision is minimal and where sediment loads are low but bed caliber high, bar growth and channel narrowing are significant adjustments at tributary junctions following impoundment. Therefore, at our sites the effects of dams on reduced competency may be more profound than on reduced sediment transport capacity, highlighting the importance of geologic and geomorphic settings in understanding fluvial responses to impoundment.

Experimental study of the free surface velocity field in an asymmetrical confluence

NASA Astrophysics Data System (ADS)

Creelle, Stephan; Mignot, Emmanuel; Schindfessel, Laurent; De Mulder, Tom

2017-04-01

The hydrodynamic behavior of open channel confluences is highly complex because of the combination of different processes that interact with each other. To gain further insights in how the velocity uniformization between the upstream channels and the downstream channel is proceeding, experiments are performed in a large scale 90 degree angled concrete confluence flume with a chamfered rectangular cross-section and a width of 0.98m. The dimensions and lay-out of the flume are representative for a prototype scale confluence in e.g. drainage and irrigation systems. In this type of engineered channels with sharp corners the separation zone is very large and thus the velocity difference between the most contracted section and the separation zone is pronounced. With the help of surface particle tracking velocimetry the velocity field is recorded from upstream of the confluence to a significant distance downstream of the confluence. The resulting data allow to analyze the evolution of the incoming flows (with a developed velocity profile) that interact with the stagnation zone and each other, causing a shear layer between the two bulk flows. Close observation of the velocity field near the stagnation zone shows that there are actually two shear layers in the vicinity of the upstream corner. Furthermore, the data reveals that the shear layer observed more downstream between the two incoming flows is actually one of the two shear layers next to the stagnation zone that continues, while the other shear layer ceases to exist. The extensive measurement domain also allows to study the shear layer between the contracted section and the separation zone. The shear layers of the stagnation zone between the incoming flows and the one between the contracted flow and separation zone are localized and parameters such as the maximum gradient, velocity difference and width of the shear layer are calculated. Analysis of these data shows that the shear layer between the incoming flows disappears quite quickly, because of the severe flow contraction that aids the flow uniformization. This is also accelerated because of a flow redistribution process that starts already upstream of the confluence, resulting in a lower than expected velocity difference over the shear layer between the bulk of the incoming flows. In contrast, the shear layer between the contracted section and the separation zone proves to be of a significantly higher order of magnitude, with large turbulent structures appearing that get transported far downstream. In conclusion, the resulting understanding of this analysis of velocity fields with a larger field of view shows that when analyzing confluence hydrodynamics, one should pay ample attention to analyze data far enough up and downstream to assess all the relevant processes.

The effects of Glen Canyon Dam operations on early life stages of rainbow trout in the Colorado River

USGS Publications Warehouse

Korman, Josh; Melis, Theodore S.

2011-01-01

The Lees Ferry reach of the Colorado River-a 16-mile segment from Glen Canyon Dam to the confluence with the Paria River-supports an important recreational rainbow trout (Oncorhynchus mykiss) fishery. In Grand Canyon, nonnative rainbow trout prey on and compete for habitat and food with native fish, such as the endangered humpback chub (Gila cypha). Experimental flow fluctuations from the dam during winter and spring 2003-5 dewatered and killed a high proportion of rainbow trout eggs in gravel spawning bars, but this mortality had no measurable effect on the abundance of juvenile fish. Flow fluctuations during summer months reduced growth of juvenile trout relative to steadier flows. A high-flow experiment in March 2008 increased both trout survival rates for early life stages and fish abundance. These findings demonstrate that Glen Canyon Dam operations directly affect the trout population in the Lees Ferry reach and could be used to regulate nonnative fish abundance to limit potential negative effects of trout on native fish in Grand Canyon.

Use of a Wiki-Based Software to Manage Research Group Activities

ERIC Educational Resources Information Center

Wang, Ting; Vezenov, Dmitri V.; Simboli, Brian

2014-01-01

This paper discusses use of the wiki software Confluence to organize research group activities and lab resources. Confluence can serve as an electronic lab notebook (ELN), as well as an information management and collaboration tool. The article provides a case study in how researchers can use wiki software in "home-grown" fashion to…

Risk Factors for Sexual Aggression in Young Men: An Expansion of the Confluence Model

PubMed Central

Abbey, Antonia; Jacques-Tiura, Angela J.; LeBreton, James M.

2011-01-01

There are many explanations for high rates of sexual aggression, with no one theory dominating the field. This study extends past research by evaluating an expanded version of the confluence model with a community sample. One hour audio computer-assisted self-interviews were completed by 470 young single men. Using structural equation analyses, delinquency, hostile masculinity, impersonal sex, and misperception of women’s sexual cues were positively and directly associated with the number of sexually aggressive acts committed. There were also indirect effects of childhood victimization, personality traits associated with subclinical levels of psychopathy, and alcohol consumption. These findings demonstrate the usefulness of the confluence model, as well as the importance of broadening this theory to include additional constructs. PMID:21678429

Increased viability of fibroblasts when pretreated with ceria nanoparticles during serum deprivation.

PubMed

Genier, Francielli S; Bizanek, Maximilian; Webster, Thomas J; Roy, Amit K

2018-01-01

Conditions of cellular stress are often the cause of cell death or dysfunction. Sustained cell stress can lead to several health complications, such as extensive inflammatory responses, tumor growth, and necrosis. To prevent disease and protect human tissue during these conditions and to avoid medication side effects, nanomaterials with unique characteristics have been applied to biological systems. This paper introduces the pretreatment in human dermal fibroblasts with cerium oxide nanoparticles during nutritional stress. For this purpose, human dermal fibroblast cells received cell culture media with concentrations of 250 µg/mL and 500 µg/mL of nano-cerium oxide before being exposed to 24, 48, and 72 hours of serum starvation. Contrast images demonstrated higher cell confluence and cell integrity in cells pretreated with ceria nanoparticles compared to untreated cells. It was confirmed by MTS assay after 72 hours of serum starvation that higher cell viability was achieved with ceria nanoparticles. The results demonstrate the potential of cerium oxide nanoparticles as protective agents during cellular starvation.

KSHV cell attachment sites revealed by ultra sensitive tyramide signal amplification (TSA) localize to membrane microdomains that are up-regulated on mitotic cells.

PubMed

Garrigues, H Jacques; Rubinchikova, Yelena E; Rose, Timothy M

2014-03-01

Cell surface structures initiating attachment of Kaposi's sarcoma-associated herpesvirus (KSHV) were characterized using purified hapten-labeled virions visualized by confocal microscopy with a sensitive fluorescent enhancement using tyramide signal amplification (TSA). KSHV attachment sites were present in specific cellular domains, including actin-based filopodia, lamellipodia, ruffled membranes, microvilli and intercellular junctions. Isolated microdomains were identified on the dorsal surface, which were heterogeneous in size with a variable distribution that depended on cellular confluence and cell cycle stage. KSHV binding domains ranged from scarce on interphase cells to dense and continuous on mitotic cells, and quantitation of bound virus revealed a significant increase on mitotic compared to interphase cells. KSHV also bound to a supranuclear domain that was distinct from microdomains in confluent and interphase cells. These results suggest that rearrangement of the cellular membrane during mitosis induces changes in cell surface receptors implicated in the initial attachment stage of KSHV entry. Copyright © 2014 Elsevier Inc. All rights reserved.

Biosensors for Cell Analysis.

PubMed

Zhou, Qing; Son, Kyungjin; Liu, Ying; Revzin, Alexander

2015-01-01

Biosensors first appeared several decades ago to address the need for monitoring physiological parameters such as oxygen or glucose in biological fluids such as blood. More recently, a new wave of biosensors has emerged in order to provide more nuanced and granular information about the composition and function of living cells. Such biosensors exist at the confluence of technology and medicine and often strive to connect cell phenotype or function to physiological or pathophysiological processes. Our review aims to describe some of the key technological aspects of biosensors being developed for cell analysis. The technological aspects covered in our review include biorecognition elements used for biosensor construction, methods for integrating cells with biosensors, approaches to single-cell analysis, and the use of nanostructured biosensors for cell analysis. Our hope is that the spectrum of possibilities for cell analysis described in this review may pique the interest of biomedical scientists and engineers and may spur new collaborations in the area of using biosensors for cell analysis.

Predicting Chronic Climate-Driven Disturbances and Their Mitigation

DOE Office of Scientific and Technical Information (OSTI.GOV)

McDowell, Nate G.; Michaletz, Sean T.; Bennett, Katrina E.

Society increasingly demands the stable provision of ecosystem resources to support our population. Resource risks from climate-driven disturbances--including drought, heat, insect outbreaks, and wildfire--are rising as a chronic state of disequilibrium results from increasing temperatures and a greater frequency of extreme events. This confluence of increased demand and risk may soon reach critical thresholds. We explain here why extreme chronic disequilibrium of ecosystem function is likely to increase dramatically across the globe, creating no-analog conditions that challenge adaptation. We also present novel mechanistic theory that combines models for disturbance mortality and metabolic scaling to link size-dependent plant mortality to changesmore » in ecosystem stocks and fluxes. Efforts must anticipate and model chronic ecosystem disequilibrium to properly prepare for resilience planning.« less

Predicting Chronic Climate-Driven Disturbances and Their Mitigation

DOE Office of Scientific and Technical Information (OSTI.GOV)

McDowell, Nate G.; Michaletz, Sean T.; Bennett, Katrina E.

Society increasingly demands the stable provision of ecosystem resources to support our population. Resource risks from climate-driven disturbances, including drought, heat, insect outbreaks, and wildfire, are growing as a chronic state of disequilibrium results from increasing temperatures and a greater frequency of extreme events. This confluence of increased demand and risk may soon reach critical thresholds. Here, we explain here why extreme chronic disequilibrium of ecosystem function is likely to increase dramatically across the globe, creating no-analog conditions that challenge adaptation. We also present novel mechanistic theory that combines models for disturbance mortality and metabolic scaling to link size-dependent plantmore » mortality to changes in ecosystem stocks and fluxes. Our efforts must anticipate and model chronic ecosystem disequilibrium to properly prepare for resilience planning.« less

Predicting Chronic Climate-Driven Disturbances and Their Mitigation

DOE PAGES

McDowell, Nate G.; Michaletz, Sean T.; Bennett, Katrina E.; ...

2017-11-13

Society increasingly demands the stable provision of ecosystem resources to support our population. Resource risks from climate-driven disturbances, including drought, heat, insect outbreaks, and wildfire, are growing as a chronic state of disequilibrium results from increasing temperatures and a greater frequency of extreme events. This confluence of increased demand and risk may soon reach critical thresholds. Here, we explain here why extreme chronic disequilibrium of ecosystem function is likely to increase dramatically across the globe, creating no-analog conditions that challenge adaptation. We also present novel mechanistic theory that combines models for disturbance mortality and metabolic scaling to link size-dependent plantmore » mortality to changes in ecosystem stocks and fluxes. Our efforts must anticipate and model chronic ecosystem disequilibrium to properly prepare for resilience planning.« less

Changes in aquatic vegetation and floodplain land cover in the Upper Mississippi and Illinois rivers (1989–2000–2010)

USGS Publications Warehouse

DeJager, Nathan R.; Rohweder, Jason J.

2017-01-01

Quantifying changes in the cover of river-floodplain systems can provide important insights into the processes that structure these landscapes as well as the potential consequences to the ecosystem services they provide. We examined net changes in 13 different aquatic and floodplain land cover classes using photo interpreted maps of the navigable portions of the Upper Mississippi River (UMR, above the confluence with the Ohio River) and Illinois River from 1989 to 2000 and from 2000 to 2010. We detected net decreases in vegetated aquatic area in nearly all river reaches from 1989 to 2000. The only river reaches that experienced a subsequent recovery of vegetated aquatic area from 2000 to 2010 were located in the northern portion of the UMR (above navigation pool 14) and two reaches in the Illinois River. Changes on the floodplain were dominated by urban development, which increased in nearly every river reach studied from 1989 to 2000. Agricultural lands declined in most river reaches from 2000 to 2010. The loss of agricultural land cover in the northern UMR was accompanied by increases in forest cover, whereas in the lower UMR and Illinois River, declines in agriculture were accompanied by increases in forest and shallow marsh communities. The changes in aquatic vegetation occupied between 5 and 20% of the total aquatic area and are likely associated with previously reported regional improvements in water clarity, while smaller (1–15% of the total floodplain area) changes in anthropogenic land cover types on the floodplain are likely driven by broad-scale socio-economic conditions.

Preparing nuclei from cells in monolayer cultures suitable for counting and for following synchronized cells through the cell cycle.

PubMed

Butler, W B

1984-08-15

A procedure is described for preparing nuclei from cells in monolayer culture so that they may be counted using an electronic particle counter. It takes only 10 to 15 min, and consists of swelling the cells in hypotonic buffer and then lysing them with the quaternary ammonium salt, ethylhexadecyldimethylammonium bromide. The cells are completely lysed, yielding a suspension of clean single nuclei which is stable, free of debris, and easily counted. The method was developed for a cell line of epithelial origin (MCF-7), which is often difficult to trypsinize to single cells. It works equally well at all cell densities up to and beyond confluence, and has been used with a variety of cells in culture, including 3T3 cells, bovine macrophages, rat mammary epithelial cells, mouse mammary tumor cell lines, and human fibroblasts. The size of the nuclei produced by this procedure is related to their DNA content, and the method is thus suitable for following cultures of synchronized cells through the cell cycle, and for performing differential counts of cells with substantial differences in DNA content.

[Differentiation of human periodontal ligament stem cells into neuron-like cells in vitro].

PubMed

Zhen, Lei; Liu, Hong-Wei

2009-02-01

To isolate and purify the human periodontal ligament stem cells (PDLSC) and investigate the differentiation potentials of PDLSC into neuron-like cells in vitro. PDLSC were isolated and cultivated. PDLSC of passage 2 was plated at a density of 5 x 10(3) per mL. At 80% confluence, the PDLSC were preinduced for 24 hours, and were subsequently replaced with an inducing medium containing certain concentration of 13-mercaptoethanal (beta-ME). After 6 hours of induction, the results were evaluated by morphological observation, immunocytochemical staining for neuron specific enolase (NSE), neurofilament (NF) and glial fibrillary acid protein (GFAP) expression and RT-PCR for NSE, NF, GFAP mRNA. Meanwhile, the uninduced PDLSC were used as a negative control. PDLSC could be differentiate into cells with typical neuronal morphology. Immunohisto-chemistry and RT-PCR confirmed that the induced cells expressed NSE and NF, two marked enzymes of neuron cell. PDLSC can be induced into neuron-like cells in vitro. PDLSC have the capability of multilineage differentiations.

The maternal to fetal transfer of immunoglobulins associated with placental lesions in sheep.

PubMed Central

Poitras, B J; Miller, R B; Wilkie, B N; Bosu, W T

1986-01-01

In this study we evaluated maternofetal transmission of immunoglobulins in ewes under conditions of altered placental morphology. Intravenous injection of human red blood cells was used to induce immunoglobulins in pregnant ewes. The hemagglutination test was used to detect antibody in maternal serum, fetal and placental fluids. Placental injury was induced by intravenous inoculation of Escherichia coli endotoxin or spores of Aspergillus fumigatus into pregnant ewes at days 99 or 100 of gestation respectively. Placental infarction, thrombosis of maternal placental vessels and variable neutrophil infiltrate characterized lesions produced by A. fumigatus. Endotoxin treated ewes developed marked placental edema, congestion, hemorrhage and focal loss of uterine epithelium. Human red blood cell agglutinating antibody was not detected in placental or fetal fluids obtained from ewes with either of the above placental lesions. Placentitis of undetermined etiology was observed in seven ewes. Two ewes had received A. fumigatus, two had received endotoxin and three were untreated ewes. Histological examination of their placentas revealed trophoblastic and endometrial epithelial necrosis and necrotizing vasculitis of the chorioallantois. Human red blood cell agglutinating antibody was detected only in the fetal and placental fluids of the seven ewes with these placental lesions. The nature of these lesions would have produced a functional confluence of the maternal and fetal circulations. Antibody transfer from dam to fetus was observed only in association with placental lesions which produced this confluence of circulations. The character of the placental lesions, rather than the mere presence of placental lesions apparently determined the transfer of immunoglobulins.(ABSTRACT TRUNCATED AT 250 WORDS) Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. PMID:3742359

78 FR 39310 - Niobrara Confluence and Ponca Bluffs Conservation Areas, NE and SD; Draft Environmental Impact...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-01

... DEPARTMENT OF THE INTERIOR Fish and Wildlife Service National Park Service [FWS-R6-R-2013-N132; FXRS1265066CCP0--134-FF06R06000] Niobrara Confluence and Ponca Bluffs Conservation Areas, NE and SD; Draft... Recreational River Headquarters, 508 East 2nd Street, Yankton, SD 57078. Mail: Nick Kaczor, USFWS, Division of...

Anatomical variations of the right hepatic veins and their relevance to surgery.

PubMed

Hribernik, Marija; de Cecchis, Lucio; Trotovsek, Blaz; Gadzijev, Eldar M; Ravnik, Dean

2003-01-01

In a morphological study of the right hepatic veins anatomical characteristics of surgical importance were looked for. 110 cadaveric human livers were prepared by the corrosion casts method. The confluence patterns of the superior right hepatic vein, the hepatocaval confluence, the accessory right hepatic veins and the anastomoses between hepatic veins in the right hemiliver were examined. Four types of the superior right hepatic vein, based on the length of its trunk and the confluence pattern of its main tributaries were determined and their frequency was calculated. Type I was found in 20%, type II in 40%, type III in 25% and type IV in 15%. Accessory right hepatic veins with a minimal caliber of 0.4 cm, which were always present in type IV, were also found in other types, all together in 27% of the casts. The tributary-free part of the superior right hepatic vein at hepatocaval confluence was longer than 1 cm in 77%. In the right hemiliver 109 anastomoses were found in 29/110 liver casts. Knowing the characteristics of different superior right hepatic vein types and of the accessory right hepatic veins may be useful in segment-oriented liver resections and in right side living donor resections.

Risk factors for sexual aggression in young men: an expansion of the confluence model.

PubMed

Abbey, Antonia; Jacques-Tiura, Angela J; LeBreton, James M

2011-01-01

There are many explanations for high rates of sexual aggression, with no one theory dominating the field. This study extends past research by evaluating an expanded version of the confluence model with a community sample. One-hour audio computer-assisted self-interviews were completed by 470 young single men. Using structural equation analyses, delinquency, hostile masculinity, impersonal sex, and misperception of women's sexual cues were positively and directly associated with the number of sexually aggressive acts committed. There were also indirect effects of childhood victimization, personality traits associated with subclinical levels of psychopathy, and alcohol consumption. These findings demonstrate the usefulness of the confluence model, as well as the importance of broadening this theory to include additional constructs. © 2011 Wiley-Liss, Inc.

Rapid isolation of choriocapillary endothelial cells by Lycopersicon esculentum-coated Dynabeads.

PubMed

Hoffmann, S; Spee, C; Murata, T; Cui, J Z; Ryan, S J; Hinton, D R

1998-10-01

In vitro studies of choroidal endothelial cells may be critical for understanding the pathogenesis of neovascularization in age-related macular degeneration, since endothelial cells from different sites are highly heterogeneous in their morphology and behavior. Isolation of choroidal endothelial cells is complicated and labor intensive because of the small size of the choroid and the difficulty of excluding contaminating cells. We describe a rapid, simplified method for the isolation of bovine choroidal endothelial cells using microdissection followed by the use of superparamagnetic beads (Dynabeads) coated with the endothelial cell-specific lectin Lycopersicon esculentum, which selectively binds to fucose residues on the endothelial cell surface. Cells bound to beads are isolated using a magnetic particle concentrator. Isolated cells grew to confluence in a monolayer with a cobblestone morphology and were shown to be endothelial cells by their greater than 95% immunoreactivity to von Willebrand factor and phagocytosis of dil-acetylated LDL. Isolated cells grew as tubes in three-dimensional cultures. This method markedly reduces the time needed for pure culture of cells and makes the in vitro study of choroidal endothelial cells practical and reproducible.

Starling resistors, autoregulation of cerebral perfusion and the pathogenesis of idiopathic intracranial hypertension.

PubMed

DE Simone, Roberto; Ranieri, Angelo; Bonavita, Vincenzo

2017-03-01

Two critical functions for the control of intracranial fluids dynamics are carried on the venous side of the perfusion circuit: the first is the avoidance of cortical veins collapse during the physiological increases of cerebrospinal fluid (CSF) pressure in which they are immersed. The second, is the generation of an abrupt venous pressure drop at the confluence of the cortical veins with the dural sinuses that is required to allow a CSF outflow rate balanced with its production. There is evidence that both of these effects are ensured by a Starling resistor mechanism (a fluid dynamic construct that governs the flow in collapsible tubes exposed to variable external pressure) acting at the confluence of cortical veins in the dural sinus. This implies that, in normal circumstances of perfusion balance, a certain degree of venous collapse physiologically occurs at the distal end of the cortical vein. This is passively modulated by the transmural pressure of the venous wall (i.e. the difference between internal blood pressure and external CSF pressure). The mechanism provides that the blood pressure of the cortical vein upstream the collapsed segment is dynamically maintained a few mmHg higher than the CSF pressure, so as to prevent their collapse during the large physiological fluctuations of the intracranial pressure. Moreover, the partial collapse of the vein confluence also generates a sharp pressure drop of the blood entering into the sinus. The CSF is drained in dural sinus through arachnoid villi proportionally to its pressure gradient with the sinus blood. The venous pressure drop between cortical veins and dural sinus is therefore needed to ensure that the CSF can leave the cranio-spinal space with the same speed with which it is produced, without having to reach a too high pressure, which would compress the cortical veins. Notably, the mechanism requires that the walls of the dural sinuses are rigid enough to avoid the collapse under the external cerebrospinal fluid pressure, and predicts that in the presence of excessively flexible dural sinuses, the system admits a second point of balance between cerebral fluid pressure and dural sinus pressure, at higher values. The second balance state is due to the triggering of a self-limiting venous collapse feedback loop between the CSF pressure, that compresses the sinus, and the subsequent increase of the dural sinus pressure, that further raises the intracranial pressure. The loop may stabilize only when the maximum stretching allowed by the venous wall is reached. Then, a new relatively stable and self-sustaining balance state is achieved, at the price of a higher CSF and dural sinus pressure values. We propose that this model is crucially involved in Idiopatic Intracranial Hypertension pathogenesis with and without papilledema, a condition that could be described as a pathological new balance state, relatively stable, between intracranial and dural venous pressure, at higher absolute values.

Bed erosion control at 60 degree river confluence using vanes

NASA Astrophysics Data System (ADS)

Wuppukondur, Ananth; Chandra, Venu

2017-04-01

Confluences are common occurrences along natural rivers. Hydrodynamics at the confluence is complex due to merging of main and lateral flows with different characteristics. Bed erosion occurs at the confluence due to turbulence and also secondary circulation induced by centrifugal action of the lateral flow. The eroded sediment poses various problems in the river ecosystem including river bank failure. Reservoirs are majorly affected due to sediment deposition which reduces storage capacity. The bed erosion also endangers stability of pipeline crossings and bridge piers. The aim of this experimental study is to check the performance of vanes in controlling bed erosion at the confluence. Experiments are performed in a 600 confluence mobile bed model with a non-uniform sediment of mean particle size d50 = 0.28mm. Discharge ratio (q=ratio of lateral flow discharge to main flow discharge) is maintained as 0.5 and 0.75 with a constant average main flow depth (h) of 5cm. Vanes of width 0.3h (1.5cm) and thickness of 1 mm are placed along the mixing layer at an angle of 150, 300 and 600(with respect to main flow) to perform the experiments. Also, two different spacing of 2h and 3h (10cm and 15cm) between the vanes are used for conducting the experiments. A digital point gauge with an accuracy of ±0.1mm is used to measure bed levels and flow depths at the confluence. An Acoustic Doppler Velocitimeter (ADV) with a frequency of 25Hz and accuracy of ±1mm/s is used to measure flow velocities. Maximum scour depth ratio Rmax, which is ratio between maximum scour depth (Ds) and flow depth (h), is used to present the experimental results.From the experiments without vanes, it is observed that the velocities are increasing along the mixing layer and Rmax=0.82 and 1.06, for q=0.5 and 0.75, respectively. The velocities reduce with vanes since roughness increases along the mixing layer. For q=0.5 and 0.75, Rmax reduces to 0.62 and 0.7 with vanes at 2h spacing, respectively. Similarly, for the same discharge ratios (q), Rmax reduces to 0.64 and 0.72 with 3h spacing between the vanes, respectively. Obstruction to the flow increases with an increase of vane angle which leads to decrease of bed erosion. Also, the bed erosion increases with an increase of spacing between the vanes. Hence, vanes placed at 600 vane angle and 2h spacing exhibit better performance.

NF2/Merlin mediates contact-dependent inhibition of EGFR mobility and internalization via cortical actomyosin.

PubMed

Chiasson-MacKenzie, Christine; Morris, Zachary S; Baca, Quentin; Morris, Brett; Coker, Joanna K; Mirchev, Rossen; Jensen, Anne E; Carey, Thomas; Stott, Shannon L; Golan, David E; McClatchey, Andrea I

2015-10-26

The proliferation of normal cells is inhibited at confluence, but the molecular basis of this phenomenon, known as contact-dependent inhibition of proliferation, is unclear. We previously identified the neurofibromatosis type 2 (NF2) tumor suppressor Merlin as a critical mediator of contact-dependent inhibition of proliferation and specifically found that Merlin inhibits the internalization of, and signaling from, the epidermal growth factor receptor (EGFR) in response to cell contact. Merlin is closely related to the membrane-cytoskeleton linking proteins Ezrin, Radixin, and Moesin, and localization of Merlin to the cortical cytoskeleton is required for contact-dependent regulation of EGFR. We show that Merlin and Ezrin are essential components of a mechanism whereby mechanical forces associated with the establishment of cell-cell junctions are transduced across the cell cortex via the cortical actomyosin cytoskeleton to control the lateral mobility and activity of EGFR, providing novel insight into how cells inhibit mitogenic signaling in response to cell contact. © 2015 Chiassson-MacKenzie et al.

Establishment and characterization of a cell line from human adenoid cystic carcinoma of the lacrimal glands and a nude mouse transplantable model.

PubMed

Lin, Tingting; Zhu, Limin; Zhou, Beiqing; Xie, Lianfeng; Lv, Jianmei; Dong, Lijie; He, Yanjin

2015-06-01

Using tissue block culture techniques, we established a new human tumor cell line derived from adenoid cystic carcinoma of the lacrimal glands (LACC-1). The LACC-1 cell line was successfully subcultured for more than 100 passages during the last two years. The outgrowth of cells was observed by day 5 after seeding, and then the cells were generated slowly. The first passage proceeded by day 32, and the classical epithelioid cell colonies formed by day 69 after inoculation. After eight passages, homogeneous epithelioid tumor cells appeared when we combined continuous passage, mechanical scraping, repeated adherence, and dissociation methods to remove the fibroblast cells. LACC-1 cells appeared as a histologically solid pattern and continuous passage culture. The population doubling time was approximately 37.1 h. LACC-1 cells appeared as an epithelioid monolayer culture on the cell culture flask and presented with a cobblestone-like appearance when they reached confluency. The nucleus was large and round with many abnormal mitoses. The nucleoplasm ratio was high. Multinucleated tumor giant cells appeared. LACC-1 cells showed a tendency to have overlapping growth without contact inhibition when the cell density continued to increase. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) showed that the LACC-1 cells were malignant tumor cells that were poorly differentiated. The surface of the LACC-1 cells exhibited affluent microvilli, protuberances and filopodia under SEM. The no. 84 generation LACC-1 cell line was inoculated subcutaneously into the subaxillary of nude mice and the tumorigenic potential was evident. The formation rate of the transplanted tumors was 100% at day 7 after inoculation. This finding showed that the LACC-1 cell line was malignant with tumorigenic ability. The xenograft tumors retained the same histological characteristics of a solid pattern as the LACC-1 original tumor after inoculation for 49 days. Under TEM observation, the xenograft tumor cells had the same ultrastructure as the LACC-1 cells. Immunohistochemical examination revealed the similarity of both cytoskeletal proteins (e.g., cytokeratin, vimentin, desmin and α-SMA) and S-100 expression in the original tumor, LACC-1 cells and xenograft tumors. Immunoreactivity of these proteins was gradually decreased in these three tissues. Reverse transcription-polymerase chain reaction demonstrated that the xenograft tumors originated from the human. Based on these results, the LACC-1 cell line provides a useful model for studying the biological characteristics of human ACC of the lacrimal glands.

Effect of brief exposure to mitomycin C on cultured human nasal mucosa fibroblasts.

PubMed

Hu, D; Sires, B S; Tong, D C; Royack, G A; Oda, D

2000-03-01

To observe the effect of mitomycin C (MMC) on cultured human nasal mucosa fibroblasts. Cultured human nasal mucosa fibroblasts were exposed to MMC (0.1-0.4 mg/ml) for 1 to 5 minutes. The viability of the fibroblasts was determined by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay; DNA fragmentation (apoptosis) by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL); apoptotic percentage by flow cytometry; and morphology by light microscopy. A portion of the fibroblasts survived the mitomycin treatment and showed evidence of regrowth within 2 to 3 days. These cells reached confluence in 5 to 7 days. The inhibition rates by MTT assay of 0.4 mg/ml MMC for 5-minute exposures was 31.3%. Dose-response effect was noted with the lower concentrations and shorter exposure times where the inhibition rates were lower (but not significantly so). DNA fragmentation was observed in fibroblasts 24 hours after MMC exposure (0.4 mg/ml) for 5 minutes compared with normal control. The apoptotic rate of fibroblasts treated by 0.4 mg/ml MMC was significantly higher than the control (p < 0.05). Short MMC exposure times have a variable cytotoxic effect and inhibit proliferation of cultured nasal mucosa fibroblasts. MMC also can induce apoptosis with a 5-minute exposure time. Therefore, it is possible that MMC has a complex effect in dacryocystorhinostomy.

Human computer confluence applied in healthcare and rehabilitation.

PubMed

Viaud-Delmon, Isabelle; Gaggioli, Andrea; Ferscha, Alois; Dunne, Stephen

2012-01-01

Human computer confluence (HCC) is an ambitious research program studying how the emerging symbiotic relation between humans and computing devices can enable radically new forms of sensing, perception, interaction, and understanding. It is an interdisciplinary field, bringing together researches from horizons as various as pervasive computing, bio-signals processing, neuroscience, electronics, robotics, virtual & augmented reality, and provides an amazing potential for applications in medicine and rehabilitation.

76 FR 53827 - Safety Zone; Big Sioux River From the Military Road Bridge North Sioux City to the Confluence of...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-30

...-AA00 Safety Zone; Big Sioux River From the Military Road Bridge North Sioux City to the Confluence of... rule extends the existing temporary safety zone on the Big Sioux River from the Military Road Bridge in... period. SUMMARY: The Coast Guard is extending the effective period for the temporary safety zone...

Field Data Collection Methods and Data Processing of the Influence of Low Momentum Ratio and the Rate of Sediment Transport Forcing on Confluence Hydrodynamics, Morphodynamics and Mixing

NASA Astrophysics Data System (ADS)

Moradi, Gelare; Cardot, Romain; Lane, Stuart; Rennie, Colin

2017-04-01

River confluences are zones where two or more rivers join and form a single channel downstream of their junction. Because of their essential role in the dynamic of fluvial networks, there has been an increase in the attention given to their hydrodynamics and morphodynamics during last three decades. Despite this increased understanding of the complex flow behavior and morphological aspects, few studies has been focused on low momentum ratio river confluences and mixing processes. As among these few studies, most of them have been driven by the mean of laboratory experiments and numerical models, a combination of field data collection and data processing is required to study the effect of low momentum ratio on flow dynamic, rive morphology and rate of mixing in river confluences. In the present poster, the flow discharge and velocity data of two upper Rhône river confluences in Switzerland, which are characterized by low momentum ratio and a varied rate of poorly sorted sediment transport is shown. The data set is mostly collected, using spatial distributed acoustic Doppler current profiling (aDcp) measurements. The morphological changes are studied using a combination of high-resolution aerial imagery data obtained by a phantom drone and acoustic bathymetric surveys. The mixing processes are investigated by measuring the surface water temperature with a thermic camera mounted on an E-bee drone [, whereas sediment pathways can be explored through the use of the 'bottom-tracking' feature of the aDcp device (not sure there will be such results at the conference time)]. These collected data is processed using a matlab code, Pix4D and visualization software. These processed data then can be used to describe the flow behavior, morphological aspects and mixing processes at river confluences characterized by low momentum ratio and to test laboratory derived conceptual models of flow processes at such junctions. The obtained results can be used under a wider range of forcing conditions to provide detailed data on the three-dimensional flow field and the morphology, to validate numerical models.

Role of contact inhibition in the regulation of receptor-mediated uptake of low density lipoprotein in cultured vascular endothelial cells.

PubMed Central

Vlodavsky, I; Fielding, P E; Fielding, C J; Gospodarowicz, D

1978-01-01

Bovine vascular endothelial cells during logarithmic growth bind, internalize, and degrade low density lipoprotein (LDL) via a receptor-mediated pathway. However, contact-inhibited (confluent) monolayers bind but do not internalize LDL. This is in contrast to aortic smooth muscle cells or endothelial cells that have lost the property of contact inhibition. These cells internalize and degrade LDL at both high and low cell densities. The LDL receptors of smooth muscle and sparse endothelial cells down-regulate in response to LDL. In contrast, normal endothelial cells at confluency show little response. When contact inhibition in endothelial monolayers was locally released by wounding, and LDL was present, only cells released from contact inhibition accumulated LDL cholesterol. In smooth muscle cells under the same conditions, the entire culture interiorized lipid. It thus appears that in endothelial cells, unlike smooth muscle cells, contact inhibition is the major factor regulating cellular uptake of LDL cholesteryl ester. Reversal of contact inhibition by wounding provides a mechanism by which the endothelium could be the primary initiator of the atherosclerotic plaque. Images PMID:203937

Iliocaval Confluence Stenting for Chronic Venous Obstructions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Graaf, Rick de, E-mail: r.de.graaf@mumc.nl; Wolf, Mark de, E-mail: markthewolf@gmail.com; Sailer, Anna M., E-mail: anni.sailer@mumc.nl

PurposeDifferent techniques have been described for stenting of venous obstructions. We report our experience with two different confluence stenting techniques to treat chronic bi-iliocaval obstructions.Materials and MethodsBetween 11/2009 and 08/2014 we treated 40 patients for chronic total bi-iliocaval obstructions. Pre-operative magnetic resonance venography showed bilateral extensive post-thrombotic scarring in common and external iliac veins as well as obstruction of the inferior vena cava (IVC). Stenting of the IVC was performed with large self-expandable stents down to the level of the iliocaval confluence. To bridge the confluence, either self-expandable stents were placed inside the IVC stent (24 patients, SECS group) ormore » high radial force balloon-expandable stents were placed at the same level (16 patients, BECS group). In both cases, bilateral iliac extensions were performed using nitinol stents.ResultsRecanalization was achieved for all patients. In 15 (38 %) patients, a hybrid procedure with endophlebectomy and arteriovenous fistula creation needed to be performed because of significant involvement of inflow vessels below the inguinal ligament. Mean follow-up was 443 ± 438 days (range 7–1683 days). For all patients, primary, assisted-primary, and secondary patency rate at 36 months were 70, 73, and 78 %, respectively. Twelve-month patency rates in the SECS group were 85, 85, and 95 % for primary, assisted-primary, and secondary patency. In the BECS group, primary patency was 100 % during a mean follow-up period of 134 ± 118 (range 29–337) days.ConclusionStenting of chronic bi-iliocaval obstruction shows relatively high patency rates at medium follow-up. Short-term patency seems to favor confluence stenting with balloon-expandable stents.« less

Enhancement of particle aggregation in the presence of organic matter during neutralization of acid drainage in a stream confluence and its effect on arsenic immobilization.

PubMed

Arce, Guillermo; Montecinos, Mauricio; Guerra, Paula; Escauriaza, Cristian; Coquery, Marina; Pastén, Pablo

2017-08-01

Acid drainage (AD) is an important environmental concern that impacts water quality. The formation of reactive Fe and Al oxyhydroxides during the neutralization of AD at river confluences is a natural attenuation process. Although it is known that organic matter (OM) can affect the aggregation of Fe and Al oxyhydroxides and the sorption of As onto their surfaces, the role of OM during the neutralization of AD at river confluences has not been studied. Field and experimental approaches were used to understand this role, using the Azufre River (pH 2) - Caracarani River (pH 8.6) confluence (northern Chile) as model system. Field measurements of organic carbon revealed a 10-15% loss of OM downstream the confluence, which was attributed to associations with Fe and Al oxyhydroxides that settle in the river bed. Laboratory mixtures of AD water with synthetic Caracarani waters under varying conditions of pH, concentration and type of OM revealed that OM promoted the aggregation of Fe oxyhydroxides without reducing As sorption, enhancing the removal of As at slightly acidic conditions (pH ∼4.5). At acidic conditions (pH ∼3), aggregation of OM - metal complexes at high OM concentrations could become the main removal mechanism. One type of OM promoted bimodal particle size distributions with larger mean sizes, possibly increasing the settling velocity of aggregates. This work contributes to a better understanding of the role of OM in AD affected basins, showing that the presence of OM during processes of neutralization of AD can enhance the removal of toxic elements. Copyright © 2017 Elsevier Ltd. All rights reserved.

Mass loading of selected major and trace elements in Lake Fork Creek near Leadville, Colorado, September-October 2001

USGS Publications Warehouse

Walton-Day, Katherine; Flynn, Jennifer L.; Kimball, Briant A.; Runkel, Robert L.

2005-01-01

A mass-loading study of Lake Fork Creek of the Arkansas River between Sugarloaf Dam and the mouth was completed in September-October 2001 to help ascertain the following: (1) variation of pH and aqueous constituent concentrations (calcium, sulfate, alkalinity, aluminum, cadmium, copper, iron, manganese, lead, and zinc) and their relation to toxicity standards along the study reach; (2) location and magnitude of sources of metal loading to Lake Fork Creek; (3) amount and locations of metal attenuation; (4) the effect of streamside wetlands on metal transport from contributing mine tunnels; and (5) the effect of organic-rich inflow from the Leadville National Fish Hatchery on water quality in Lake Fork Creek. The study was done in cooperation with the Bureau of Land Management, U.S. Department of Agriculture Forest Service, and U.S. Fish and Wildlife Service. Constituent concentrations and pH showed variable patterns over the study reach. Hardness-based acute and chronic toxicity standards were exceeded for some inflows and some constituents. However, stream concentrations did not exceed standards except for zinc starting in the upper parts of the study reach and extending to just downstream from the inflow from the Leadville National Fish Hatchery. Dilution from that inflow lowered stream zinc concentrations to less than acute and chronic toxicity standards. The uppermost 800 meters of the study reach that contained inflow from the Bartlett, Dinero, and Nelson mine tunnels and the Dinero wetland was the greatest source of loading for manganese and zinc. A middle section of the study reach that extended approximately 2 kilometers upstream from the National Fish Hatchery inflow to just downstream from that inflow was the largest source of aluminum, copper, iron, and lead loading. The loading was partially from the National Fish Hatchery inflow but also from unknown sources upstream from that inflow, possibly ground water. The largest sources for calcium and sulfate load to the stream were the parts of the study reach containing inflow from the tribu-taries Halfmoon Creek (calcium) and Willow Creek (sulfate). The Arkansas River and its tributaries upstream from Lake Fork Creek were the source of most of the calcium (70 percent), sulfate (82 percent), manganese (77 percent), lead (78 percent), and zinc (95 percent) loads in the Arkansas River downstream from the Lake Fork confluence. In contrast, Lake Fork Creek was the major source of aluminum (68 percent), copper (65 percent), and iron (87 percent) loads to the Arkansas River downstream from the confluence. Attenuation was not important for calcium, sulfate, or iron. However, other metals loads were reduced up to 81 percent over the study reach (aluminum, 25 percent; copper, 20 percent; manganese, 81 percent; lead, 30 percent; zinc, 72 percent). Metal attenuation in the stream occurred primarily in three locations (1) the irrigation diversion ditch; (2) the beaver pond complex extending from upstream from the Colorado Gulch inflow to just downstream from that inflow; and (3) the stream reach that included the inflow from Willow Creek. The most likely attenuation mechanism is precipitation of metal oxides and hydroxides (primarily manganese), and sorption or coprecipitation of trace elements with the precipitating phase. A mass-balance calculation indicated that the wetland between the Dinero Tunnel and Lake Fork Creek removed iron, had little effect on zinc mass load, and was a source for, or was releasing, aluminum and manganese. In contrast, the wetland that occurred between the Siwatch Tunnel and Lake Fork Creek removed aluminum, iron, manganese, and zinc from the tunnel drainage before it entered the creek. Inflow from the National Fish Hatchery increased dissolved organic carbon concentrations in Lake Fork Creek and slightly changed the composition of the dissolved organic carbon. However, dissolved organic carbon loads increased in the stream reach downs

10. WHITNEY'S FLUME AND VIEW OF THE CONFLUENCE OF TONTO ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

10. WHITNEY'S FLUME AND VIEW OF THE CONFLUENCE OF TONTO CREEK AND THE SALT RIVER. AREA SHOWN IS PRESENTLY UNDER WATER. TONTO CREEK FLOWS FROM BACKGROUND CENTER TO LEFT, AND THE SALT RIVER FLOWS FROM RIGHT TO LEFT IN THE PHOTO. DAM IS LOCATED OFF THE PHOTO TO THE LEFT Photographer: Walter J. Lubken, March 3, 1906 - Roosevelt Power Canal & Diversion Dam, Parallels Salt River, Roosevelt, Gila County, AZ

Humic substances and trace metals associated with Fe and Al oxides deposited in an acidic mountain stream

USGS Publications Warehouse

McKnight, Diane M.; Wershaw, R. L.; Bencala, K.E.; Zellweger, G.W.; Feder, G.L.

1992-01-01

Hydrous iron and aluminum oxides are deposited on the streambed in the confluence of the Snake River and Deer Creek, two streams in the Colorado Rocky Mountains. The Snake River is acidic and has high concentrations of dissolved Fe and Al. These metals precipitate at the confluence with the pristine, neutral pH, Deer Creek because of the greater pH (4.5-6.0) in the confluence. The composition of the deposited oxides changes consistently with distance downstream, with the most upstream oxide samples having the greatest Fe and organic carbon content. Fulvic acid accounts for most of the organic content of the oxides. Results indicate that streambed oxides in the confluence are not saturated with respect to their capacity to sorb dissolved humic substances from streamwater. The contents of several trace metals (Mn, Zn, Cu, Pb, Ni and Co) also decrease with distance downstream and are correlated with both the Fe and organic carbon contents. Strong metal-binding sites associated with the sorbed fulvic acid are more than sufficient to account for the trace metal content of the oxides. Complexation of trace metals by sorbed fulvic acid may explain the observed downstream decrease in trace metal content.

The influence of Dworshak Dam on epilithic community metabolism in the Clearwater River, U.S.A.

USGS Publications Warehouse

Munn, M.D.; Brusven, M.A.

2004-01-01

Epilithic community metabolism was determined on a seasonal basis over two years in nonregulated and regulated reaches of the Clearwater River in northern Idaho, U.S.A. Metabolism was estimated using three, 12-liter recirculating chambers and the dissolved oxygen method, with parameters expressed as g O2 m−2 d−1. In the nonregulated reach above the reservoir, gross community productivity (GCP) ranged from 0.8 to 3.2, community respiration (CR24) from 0.3 to 1.2, and production/respiration (P/R) ratios from 1.2 to 3.3. Epilithic metabolism in the regulated reach immediately below the dam increased sharply; GCP ranged from 4.2 to 25.5, CR24 from 1.9 to 9.7, and P/R ratios from 1.4 to 5.7. Increased primary production and respiration in the regulated reach was a result of extensive growth of an aquatic moss (Fontanalis neo-mexicanus). The influence of the dam on epilithic community metabolism was mitigated 2.5 km downstream of the dam due to the regulated North Fork of the Clearwater River (NFCR) merging with the larger, nonregulated Clearwater River. While the regulated Clearwater River below the confluence was somewhat affected by the regulated NFCR flows upstream, metabolism was similar to that found above the reservoir (GCP = 1.2 – 2.6, CR24 = 0.6 – 1.3, and P/R = 1.4 – 2.2). This study demonstrates that while Dworshak Dam has altered both primary production and respiration directly below the dam, the placement of the dam only 2.5 km upstream from a nonregulated reach greatly mitigates its effects on stream metabolism downstream.

Channel adjustements over the last century of the Moldova River, Romania

NASA Astrophysics Data System (ADS)

Chiriloaei, F. A.; Radoane, M.; Radoane, N.

2012-04-01

Moldova River is a 205 km long river, right tributary of the Siret River, with the confluence close to Roman city. The most important tributaries are Moldoviţa, Suha Mică, Suha Mare, Râsca, Ozana and Topolita Rivers. The drainage basin area is 4316 km2, a discharge of about 32.8 m3/s and is superimposed on four lithostratigraphic units: the crystalline Mesozoic unit, the flysch unit, the molasse unit and the platform unit. The study reach is 110 km long and is located on the external part of the Eastern-Carpathians at the contact with the Moldavian Plateau. This contact is characterised by a piedmont zone. The high rate of alluviation in this piedmont plain had a decisive role on the spatial and temporal evolution of fluvial forms and processes, expressed in the morphology of alluvial terraces in the valley bottom, the morphology of the active channel, the spatial distribution of bars and secondary channels and lateral migration rates. The morphology of the river in the study reach is dominated by braided and wandering channel patterns. The present (2005) active channel width ranges between 700 and 1000 m. Planform changes of river features over the last 100 years were analyzed on three historical maps (1910, 1960, and 1980) and one orthophoto (2005). Channel width average has significantly decreased in the last century. The Moldova river channel width suffered a strong narrowing, approximately 76% (from 1910 to 2005). So, from a channel width about more than 1200 m, it decreased at about 300 m. We can note two phase of narrowing: a first one - stronger, of 56%, untill 1960 (a reduction of active channel width for about 10.5 m/year) and a second phase, of 35%, after 1960, with a narrowing rate for about 8.8 m/an. The historical trend of braiding index shows a remarkable decrease in the last 50 - 60 years, from 3.2 in 1960, at 2.6 in 1980 and 2.0, in 2005, for all the extra-Carpathian study reach (110 km). There are different situations at local scale, shown using the sectorial analysis. The rates of channel incision were estimated by analyzing the lowest annual stage of the Moldova River, in 4 gauging stations. Along the river, the incision rate (absolute values) increase from - 80 cm (at the exit of mountain area) to - 2.60 m at the confluence with Siret River. In terms of human impact, in-channel gravel mining and local embankments near bridges and towns are the most important direct interventions along the channel. Besides the direct effect of channelization on channel morphology, the major effect of human actions was on sediment regime. A significant decrease of in-channel sediment supply was determined by gravel mining. On the other hand, channel-forming discharges did not undergo significant changes in most of the study streams. It is obvious that, channel with narrowed considerably, river flow concentrated, therefore, the number of channels (links) reduced (from maximum 10, in 1960, to maximum 8 in 1980 and approximately 5, in 2005). In this period, some reaches changed their typology from braided reaches to wandering reaches.

The Use of Multidimensional Image-Based Analysis to Accurately Monitor Cell Growth in 3D Bioreactor Culture

PubMed Central

Baradez, Marc-Olivier; Marshall, Damian

2011-01-01

The transition from traditional culture methods towards bioreactor based bioprocessing to produce cells in commercially viable quantities for cell therapy applications requires the development of robust methods to ensure the quality of the cells produced. Standard methods for measuring cell quality parameters such as viability provide only limited information making process monitoring and optimisation difficult. Here we describe a 3D image-based approach to develop cell distribution maps which can be used to simultaneously measure the number, confluency and morphology of cells attached to microcarriers in a stirred tank bioreactor. The accuracy of the cell distribution measurements is validated using in silico modelling of synthetic image datasets and is shown to have an accuracy >90%. Using the cell distribution mapping process and principal component analysis we show how cell growth can be quantitatively monitored over a 13 day bioreactor culture period and how changes to manufacture processes such as initial cell seeding density can significantly influence cell morphology and the rate at which cells are produced. Taken together, these results demonstrate how image-based analysis can be incorporated in cell quality control processes facilitating the transition towards bioreactor based manufacture for clinical grade cells. PMID:22028809

The use of multidimensional image-based analysis to accurately monitor cell growth in 3D bioreactor culture.

PubMed

Baradez, Marc-Olivier; Marshall, Damian

2011-01-01

The transition from traditional culture methods towards bioreactor based bioprocessing to produce cells in commercially viable quantities for cell therapy applications requires the development of robust methods to ensure the quality of the cells produced. Standard methods for measuring cell quality parameters such as viability provide only limited information making process monitoring and optimisation difficult. Here we describe a 3D image-based approach to develop cell distribution maps which can be used to simultaneously measure the number, confluency and morphology of cells attached to microcarriers in a stirred tank bioreactor. The accuracy of the cell distribution measurements is validated using in silico modelling of synthetic image datasets and is shown to have an accuracy >90%. Using the cell distribution mapping process and principal component analysis we show how cell growth can be quantitatively monitored over a 13 day bioreactor culture period and how changes to manufacture processes such as initial cell seeding density can significantly influence cell morphology and the rate at which cells are produced. Taken together, these results demonstrate how image-based analysis can be incorporated in cell quality control processes facilitating the transition towards bioreactor based manufacture for clinical grade cells.

Transforming capacity of two novel genes JS-1 and JS-2 located in chromosome 5p and their overexpression in human esophageal squamous cell carcinoma.

PubMed

Fatima, Sarwat; Chui, Chung H; Tang, Wing K; Hui, Kin S; Au, Ho W; Li, Wing Y; Wong, Mei M; Cheung, Filly; Tsao, S W; Lam, King Y; Beh, Philip S L; Wong, John; Law, Simon; Srivastava, Gopesh; Ho, Kwok P; Chan, Albert S C; Tang, Johnny C O

2006-01-01

Esophageal squamous cell carcinoma (ESCC) has a high mortality rate and geographic differences in incidence. Previous studies of comparative genomic hybridization (CGH) showed that chromosomal 5p is frequently amplified in cell lines and primary ESCC of Hong Kong Chinese origin. In this report, attempt was made to study two novel genes, named as JS-1 and JS-2, which are located in chromosome 5p15.2 and are 5' upstream to delta catenin for their roles in molecular pathogenesis of ESCC. Eleven cell lines, 27 primary ESCC cases and multiple human tissue cDNA panels (MTC) of digestive system were studied for the expression level of JS-1 and JS-2 by RT-PCR. The full-length cDNA sequences of JS-1 and JS-2 were determined from a non-tumor esophageal epithelial cell line by 3' and 5' rapid amplification of cDNA ends (RACE). The transforming capacity of JS-1 and JS-2 was also investigated by transfecting NIH 3T3 cells with the expression vector pcDNA3.1(-) cloned with the full coding sequences and it was followed by the study of foci formation of the transfected cells under confluence growth and the anchorage-independent growth in soft agar. Forty-five percent (5/11) and 18% (2/11) of the ESCC cell lines showed overexpression of JS-1 and JS-2 respectively, while 55% (15/27) and 14% (3/22) primary ESCC cases showed overexpression of JS-1 and JS-2 respectively. JS-1 overexpression was most common in patients with stage II ESCC (6/27; 22%) whereas JS-2 was only overexpressed in a dysplastic lesion (1/22; 4%) and stage III tumors (2/22; 9%). The expression levels of JS-1 and JS-2 are both low in normal esophageal tissues. Overexpression of JS-1 in NIH 3T3 cells caused foci formation in confluence growth and colony formation in soft agar but not for JS-2. A high grade sarcoma was formed in the athymic nude mice when NIH 3T3 cells overexpressing JS-1 were injected subcutaneously. Our results thus indicate that the frequent overexpression of JS-1 in ESCC and its transforming capacity in normal cells may play a critical role in the molecular pathogenesis of ESCC. The present study also forms the ground work for further identification of novel mechanisms of molecular carcinogenesis in ESCC and other cancers.

The INF (Intermediate-Range Nuclear Forces) Controversy: A Confluence of Foreign and Domestic Interests.

DTIC Science & Technology

1986-09-01

AD-R175 303 THE INF (INTERMEDIRTE-RANSE NUCLEAR FORCES) 1/2 CONTROVERSY: A CONFLUENCE OF FOREIGN AND DOMESTIC INTERESTS(U) NAVAL POSTGRADUATE SCHOOL...on reverse if necessary and identify by block number) F:ELD I GROUP SUB-GROUP INF (Intermediate-Range Nuclear Forces, NATO, European Nuclear Capability...the United Kingdom, and several non- nuclear members of 14ATO are examined and analyzed. The analysis is concerned with alliance and transnational

Kankakee River Basin: Evaluation of Sediment Management Strategies

DTIC Science & Technology

2013-09-01

extends from South Bend, Indiana, to its confluence with the Illinois River near Wilmington, Illinois. The river has a 5,165- square-mile drainage area and...confluence with the Illinois River near Wilmington, IL (Figure 1.1). It has a 5,165-square-mile drainage area and a river length of approximately 150 miles...Yellow River drainage area is overlain by sand-sized sediment. The Rock Island, St. Louis, Chicago, and Detroit Districts collaborated to produce the

The influence of tributary flow density differences on the hydrodynamic behavior of a confluent meander bend and implications for flow mixing

NASA Astrophysics Data System (ADS)

Herrero, Horacio S.; Díaz Lozada, José M.; García, Carlos M.; Szupiany, Ricardo N.; Best, Jim; Pagot, Mariana

2018-03-01

The goal of this study is to evaluate the influence of tributary flow density differences on hydrodynamics and mixing at a confluent meander bend. A detailed field characterization is performed using an Acoustic Doppler Current Profiler (ADCP) for quantification of the 3D flow field, flow discharge and bathymetry, as well as CTD measurements (conductivity, temperature, depth) to characterize the patterns of mixing. Satellite images of the confluence taken at complementary times to the field surveys were analyzed to evaluate the confluence hydrodynamics at different flow conditions. The results illustrate the differences in hydrodynamics and mixing length in relation to confluences with equal density tributaries. At low-density differences, and higher discharge ratio (Qr) between the two rivers, the flow is similar to equi-density confluent meander bends. In contrast, at high-density differences (low Qr), the tributary flow is confined to near the confluence but the density difference causes the flow to move across channel. In this case, the density difference causes the lateral spread of the tributary flow to be greater than at a greater Qr when the density difference is less. These results illustrate the potential importance of density differences between tributaries in determining the rate and spatial extent of mixing and sediment dispersal at confluent meander bends.

Water mass interaction in the confluence zone of the Daning River and the Yangtze River--a driving force for algal growth in the Three Gorges Reservoir.

PubMed

Holbach, Andreas; Wang, Lijing; Chen, Hao; Hu, Wei; Schleicher, Nina; Zheng, Binghui; Norra, Stefan

2013-10-01

Increasing eutrophication and algal bloom events in the Yangtze River Three Gorges Reservoir, China, are widely discussed with regard to changed hydrodynamics and nutrient transport and distribution processes. Insights into water exchange and interaction processes between water masses related to large-scale water level fluctuations in the reservoir are crucial to understand water quality and eutrophication dynamics. Therefore, confluence zones of tributaries with the Yangtze River main stream are dedicated key interfaces. In this study, water quality data were recorded in situ and on-line in varying depths with the MINIBAT towed underwater multi-sensor system in the confluence zone of the Daning River and the Yangtze River close to Wushan City during 1 week in August 2011. Geostatistical evaluation of the water quality data was performed, and results were compared to phosphorus contents of selective water samples. The strongly rising water level throughout the measurement period caused Yangtze River water masses to flow upstream into the tributary and supply their higher nutrient and particulate loads into the tributary water body. Rapid algal growth and sedimentation occurred immediately when hydrodynamic conditions in the confluence zone became more serene again. Consequently, water from the Yangtze River main stream can play a key role in providing nutrients to the algal bloom stricken water bodies of its tributaries.

Effects of recreational flow releases on natural resources of the Indian and Hudson Rivers in the Central Adirondack Mountains, New York, 2004-06

USGS Publications Warehouse

Baldigo, Barry P.; Mulvihill, C.I.; Ernst, A.G.; Boisvert, B.A.

2011-01-01

The U.S. Geological Survey (USGS), the New York State Department of Environmental Conservation (NYSDEC), and Cornell University carried out a cooperative 2-year study from the fall of 2004 through the fall of 2006 to characterize the potential effects of recreational-flow releases from Lake Abanakee on natural resources in the Indian and Hudson Rivers. Researchers gathered baseline information on hydrology, temperature, habitat, nearshore wetlands, and macroinvertebrate and fish communities and assessed the behavior and thermoregulation of stocked brown trout in study reaches from both rivers and from a control river. The effects of recreational-flow releases (releases) were assessed by comparing data from affected reaches with data from the same reaches during nonrelease days, control reaches in a nearby run-of-the-river system (the Cedar River), and one reach in the Hudson River upstream from the confluence with the Indian River. A streamgage downstream from Lake Abanakee transmitted data by satellite from November 2004 to November 2006; these data were used as the basis for developing a rating curve that was used to estimate discharges for the study period. River habitat at most study reaches was delineated by using Global Positioning System and ArcMap software on a handheld computer, and wetlands were mapped by ground-based measurements of length, width, and areal density. River temperature in the Indian and Hudson Rivers was monitored continuously at eight sites during June through September of 2005 and 2006; temperature was mapped in 2005 by remote imaging made possible through collaboration with the Rochester Institute of Technology. Fish communities at all study reaches were surveyed and characterized through quantitative, nearshore electrofishing surveys. Macroinvertebrate communities in all study reaches were sampled using the traveling-kick method and characterized using standard indices. Radio telemetry was used to track the movement and persistence of stocked brown trout (implanted with temperature-sensitive transmitters) in the Indian and Hudson Rivers during the summer of 2005 and in all three rivers during the summer of 2006. The releases had little effect on river temperatures, but increased discharges by about one order of magnitude. Regardless of the releases, river temperatures at all study sites commonly exceeded the threshold known to be stressful to brown trout. At most sites, mean and median water temperatures on release days were not significantly different, or slightly lower, than water temperatures on nonrelease days. Most differences were very small and, thus, were probably not biologically meaningful. The releases generally increased the total surface area of fast-water habitat (rapids, runs, and riffles) and decreased the total surface area of slow-water habitat (pools, glides, backwater areas, and side channels). The total surface areas of wetlands bordering the Indian River were substantially smaller than the surface areas bordering the Cedar River; however, no channel geomorphology or watershed soil and topographic data were assessed to determine whether the releases or other factors were mainly responsible for observed differences. Results from surveys of resident biota indicate that the releases generally had a limited effect on fish and macroinvertebrate communities in the Indian River and had no effect on communities in the Hudson River. Compared to fish data from Cedar River control sites, the impoundment appeared to reduce total density, biomass, and richness in the Indian River at the first site downstream from Lake Abanakee, moderately reduce the indexes at the other two sites on the Indian River, and slightly reduce the indexes at the first Hudson River site downstream from the confluence with the Indian River. The densities of individual fish populations at all Indian River sites were also reduced, but related effects on fish populations in the Hudson River were less evident. Altho

Hydrology, vegetation, and soils of riverine and tidal floodplain forests of the lower Suwannee River, Florida, and potential impacts of flow reductions

USGS Publications Warehouse

Light, Helen M.; Darst, Melanie R.; Lewis, Lori J.; Howell, David A.

2002-01-01

A study relating hydrologic conditions, soils, and vegetation of floodplain forests to river flow was conducted in the lower Suwannee River, Florida, from 1996 to 2000. The study was done by the U.S. Geological Survey in cooperation with the Suwannee River Water Management District to help determine the minimum flows and levels required for wetlands protection. The study area included forests within the 10-year floodplain of the Suwannee River from its confluence with the Santa Fe River to the tree line (lower limit of forests) near the Gulf of Mexico, and covered 18,600 hectares (ha) of forests, 75 percent of which were wetlands and 25 percent uplands. The floodplain was divided into three reaches, riverine, upper tidal, and lower tidal, based on changes in hydrology, vegetation, and soils with proximity to the coast. The Suwannee River is the second largest river in Florida in terms of average discharge. Median flow at the confluence of the Suwannee and Santa Fe Rivers is approximately 181 cubic meters per second (m3/s) or 6,480 cubic feet per second (ft3/s) (1933-99). At the upper end of the riverine reach, river stages are unaffected by tides and have a typical annual range of 4.1 meters (m). Tides affect river stages at low and medium flows in the upper tidal reach, and at all flows in the lower tidal reach. Median tidal range at the mouth of the Suwannee River is about 1 m. Salinity of river water in the lower tidal reach increases with decreasing flow and proximity to the Gulf of Mexico. Vertically averaged salinity in the river near the tree line is typically about 5 parts per thousand at medium flow. Land-surface elevation and topographic relief in the floodplain decrease with proximity to the coast. Elevations range from 4.1 to 7.3 m above sea level at the most upstream riverine transect and from 0.3 to 1.3 m above sea level on lower tidal transects. Surface soils in the riverine reach are predominantly mineral and dry soon after floods recede except in swamps. Surface soils in upper and lower tidal reaches are predominantly organic, saturated mucks. In the downstream part of the lower tidal reach, conductivities of surface soils are high enough (greater than 4 milli-mhos per centimeter) to exclude many tree species that are intolerant of salinity. Species richness of canopy and subcanopy plants in wetland forests in the lower Suwannee River is high compared to other river floodplains in North America. A total of 77 tree, shrub, and woody vine species were identified in the canopy and subcanopy of floodplain wetland forests (n = 8,376). Fourteen specific forest types were mapped using digitized aerial photographs, defined from vegetative sampling, and described in terms of plant species composition. For discussion purposes, some specific wetland types were combined, resulting in three general wetland forest types for each reach. Riverine high bottomland hardwoods have higher canopy species richness than all other forest types (40-42 species), with Quercus virginiana the most important canopy tree by basal area. The canopy composition of riverine low bottomland hardwoods is dominated by five species with Quercus laurifolia the most important by basal area. Riverine swamps occur in the lowest and wettest areas with Taxodium distichum the most important canopy species by basal area. Upper tidal bottomland hardwoods are differentiated from riverine forests by the presence of Sabal palmetto in the canopy. Upper tidal mixed forests and swamps are differentiated from riverine forests, in part, by the presence of Fraxinus profunda in the canopy. Nyssa aquatica, the most important canopy species by basal area in upper tidal swamps, is absent from most forests in the lower tidal reach where its distribution is probably restricted by salinity. Hydric hammocks, a wetland type that is rare outside of Florida, are found in the lower tidal reach and are flooded every 1-2 years by either storm surge or river floods. Lowe

Inflammatory Cytokines Induce Ubiquitination and Loss of the Prostate Suppressor Protein NKX3.1

DTIC Science & Technology

2007-10-01

Lipoproteins ( LDL ), these receptors mediate the innate immune response against bacterial infection.(44) MSR1 maps to chromosome 8p22 and is shown to...with 5% FBS and grown to 80% confluency. Cells were treated with 40 ng/ml TNF-α for 6 hours and total RNA was extracted using a RNeasy mini kit (Qiagen...NKX3.1 and β-actin primers were added at a final concentration of 0.6µM to 1.5 µg template RNA . Qiagen OneStep RT-PCR enzyme mix was used for

The Morphodynamic Signature of Rivers in the Ucamara Depression: A Habitat for Formative Rivers and the Scavenger Meandering Channels they Feed

NASA Astrophysics Data System (ADS)

Abad, J. D.; Escobar, C.; Shan, J.

2017-12-01

The Pacaya Samiria National Reserve, located in Loreto, Peru, is a region of incomparable biodiversity resulting from the consistent annual climate patterns (little seasonal variability), and more importantly, the dynamics of the freshwater rivers that surround and traverse it. The Ucamara Depression, where the Pacaya Samiria National Reserve is located, presently has a myriad of active and abandoned fluvial landforms. The exploration of the geologic and tectonic history that fabricated this exceptional fluvial system is the foundation for researching and understanding further phenomena of the region. The interpretation of the history of the geologic events that occurred to form this region and the inspection of the river belts, or areas of active river migration, of these fluvial landforms, facilitate the understanding of 1) how the Ucayali and Maranon rivers interact with one another and with the streams and bodies of water in the Ucamara Depression, 2) the role of wetlands, hydrodynamics, and sediment transport mechanisms in the movement of rivers and the extent of mixing before the rivers reach their confluence, and 3) how the water chemistry, flooding, and sediment transport processes of rivers create an environment capable of fostering an unimaginable array of life and how changes in these processes affect the flora and fauna that inhabit the region. This study will discuss field measurements (hydrodynamic and bed morphodynamic) and remote sensing analysis of scavenger meandering channels (Pacaya and Samiria) and discuss confluence dynamics of the two tributaries that form the Amazon River. Morphometric parameters show that these meandering rivers do not achieve typical planform-based conditions.

Simulation of Flood Profiles for Catoma Creek near Montgomery, Alabama, 2008

USGS Publications Warehouse

Lee, K.G.; Hedgecock, T.S.

2008-01-01

A one-dimensional step-backwater model was used to simulate flooding conditions for Catoma Creek near Montgomery, Alabama. A peak flow of 50,000 cubic feet per second was computed by the U.S. Geological Survey for the March 1990 flood at the Norman Bridge Road gaging station. Using this estimated peak flow, flood-plain surveys with associated roughness coefficients, and surveyed high-water marks for the March 1990 flood, a flow model was calibrated to closely match the known event. The calibrated model then was used to simulate flooding for the 10-, 50-, 100-, and 500-year recurrence-interval floods. The 100-year flood stage for the Alabama River also was computed in the vicinity of the Catoma Creek confluence using observed high-water profiles from the 1979 and 1990 floods and gaging-station data. The results indicate that the 100-year flood profile for Catoma Creek within the 15-mile study reach is about 2.5 feet higher, on average, than the profile published by the Federal Emergency Management Agency. The maximum and minimum differences are 6.0 feet and 0.8 foot, respectively. All water-surface elevations computed for the 100-year flood are higher than those published by the Federal Emergency Management Agency. The 100-year flood stage computed for the Alabama River in the vicinity of the Catoma Creek confluence was about 4.5 feet lower than the elevation published by the Federal Emergency Management Agency. The results of this study provide the community with flood-profile information that can be used for flood-plain mitigation, future development, and safety plans for the city.

Comparative 1D and 3D numerical investigation of open-channel junction flows and energy losses

NASA Astrophysics Data System (ADS)

Luo, Hao; Fytanidis, Dimitrios K.; Schmidt, Arthur R.; García, Marcelo H.

2018-07-01

The complexity of open channel confluences stems from flow mixing, secondary circulation, post-confluence flow separation, contraction and backwater effects. These effects in turn result in a large number of parameters required to adequately quantify the junction induced hydraulic resistance and describe mean flow pattern and turbulent flow structures due to flow merging. The recent development in computing power advances the application of 3D Computational Fluid Dynamics (CFD) codes to visualize and understand the Confluence Hydrodynamic Zone (CHZ). Nevertheless, 1D approaches remain the mainstay in large drainage network or waterway system modeling considering computational efficiency and data availability. This paper presents (i) a modified 1D nonlinear dynamic model; (ii) a fully 3D non-hydrostatic, Reynolds-averaged Navier-Stokes Equations (RANS)-based, Computational Fluid Dynamics (CFD) model; (iii) an analysis of changing confluence hydrodynamics and 3D turbulent flow structure under various controls; (iv) a comparison of flow features (i.e. upstream water depths, energy losses and post-confluence contraction) predicted by 1D and 3D models; and (v) parameterization of 3D flow characteristics in 1D modeling through the computation of correction coefficients associated with contraction, energy and momentum. The present comprehensive 3D numerical investigation highlights the driving mechanisms for junction induced energy losses. Moreover, the comparative 1D and 3D study quantifies the deviation of 1D approximations and associated underlying assumptions from the 'true' resultant flow field. The study may also shed light on improving the accuracy of the 1D large network modeling through the parameterization of the complex 3D feature of the flow field and correction of interior boundary conditions at junctions of larger angles and/or with substantial lateral inflows. Moreover, the enclosed numerical investigations may enhance the understanding of the primary mechanisms contributing to hydraulic structure induced turbulent flow behavior and increased hydraulic resistance.

Hybridization and cytonuclear associations among native westslope cutthroat trout, introduced rainbow trout, and their hybrids within the Stehekin River drainage, North Cascades National Park

USGS Publications Warehouse

Ostberg, C.O.; Rodriguez, R.J.

2006-01-01

Historic introductions of nonnative rainbow trout Oncorhynchus mykiss into the native habitats of cutthroat trout O. clarkii have impacted cutthroat trout populations through introgressive hybridization, creating challenges and concerns for cutthroat trout conservation. We examined the effects of rainbow trout introductions on the native westslope cutthroat trout O. c. lewisii within the Stehekin River drainage, North Cascades National Park, Washington, by analyzing 1,763 salmonid DNA samples from 18 locations with nine diagnostic nuclear DNA markers and one diagnostic mitochondrial DNA (mtDNA) marker. Pure westslope cutthroat trout populations only occurred above upstream migration barriers in the Stehekin River and Park Creek. Two categories of rainbow trout admixture were observed: (1) less than 10% within the Stehekin River drainage above the Bridge Creek confluence and the middle and upper Bridge Creek drainage and (2) greater than 30% within the Stehekin River below the Bridge Creek confluence and in lower Bridge Creek. Hybrid indices and multilocus genotypes revealed an absence of rainbow trout and reduced hybrid diversity within the Stehekin River above the Bridge Creek confluence relative to hybrid diversity in the Stehekin River below the confluence and within lower Bridge Creek. Cytonuclear disequilibrium statistics revealed assortative mating between westslope cutthroat and rainbow trout but not among hybrids within the same locations. This suggests that a randomly mating hybrid swarm does not currently exist. However, continual migration of parental genotypes into the study location could also create significant cytonuclear disequilibria. The Stehekin River represents a novel and unique example of a dynamic hybridization zone where the invasion of rainbow trout alleles into the Stehekin River westslope cutthroat trout population above the Bridge Creek confluence appears to be impeded, suggesting that divergent ecological or evolutionary mechanisms promote the population structure within the Stehekin River drainage, depending upon location.

Geomorphic Function and Restoration Potential of Spring Creeks in Southeastern Idaho: Analysis and Communication

NASA Astrophysics Data System (ADS)

Hanrahan, T. P.; Hill, Z.; Levell, A.; Maguire, T.; Risso, D.

2014-12-01

A large wetland and floodplain complex adjacent to the Snake River in southeastern Idaho, USA, encompasses numerous spring-fed creeks that originate on the floodplain and discharge at their confluence with the Snake River and American Falls Reservoir. Resource managers are implementing a program to restore these spring creeks for the recovery of Yellowstone cutthroat trout and ecosystem health. Our objectives were to evaluate the physical characteristics of these spring creeks, develop a conceptual model of their geomorphic function, compare the restoration potential of individual reaches, and communicate our findings to a broad audience of resource managers and regional stakeholders in order to foster restoration planning. A geomorphic assessment along 38 km of three spring creeks was completed by collecting data at several transects within distinct geomorphic reaches, and by collecting data continuously throughout all reaches. These data were summarized in a GIS database and used to quantify the overall geomorphic functioning of each reach. The geomorphic functional scores were scaled from 0% (non-functional) to 100% (fully functional). Among all three spring creeks, geomorphic function ranged from 29% to 63%, with bank conditions and riparian vegetation being the primary causes of overall channel degradation. Results from the geomorphic assessment fostered the development of a conceptual model for spring creek function, whereby degraded bank conditions represent the primary controlling factor of decreased geomorphic function and fish habitat quality. The reach-based geomorphic functional scoring provides an indicator of relative restoration potential for each reach, and is one of the factors used in determining site-specific priorities for protecting, enhancing, and restoring spring creeks on the Fort Hall Bottoms. The study results, conceptual model and restoration strategy were communicated to resource managers and regional stakeholders through a graphically-rich, large format atlas document. Presentation of hard copy and electronic versions of maps and infographics fostered a high level of engagement among those interested in restoring these spring creek systems.

Dynamics of the Brazil-Malvinas Confluence: Energy Conversions

NASA Astrophysics Data System (ADS)

Francisco, C. P. F.; da Silveira, I. C. A.; Campos, E. J. D.

2011-03-01

In this work, we investigated the mesoscale dynamics of the Brazil-Malvinas Confluence (BMC) region. Particularly, we were interested in the role of geophysical instability in the formation and development of the mesoscale features commonly observed in this region. We dynamically analyzed the results of numerical simulations of the BMC region conducted with 'Hybrid Coordinate Ocean Model' (HYCOM). We quantified the effect of barotropic and baroclinic energy conversions in the modeled flow and showed the dominance of the latter in the region.

Hydroxyapatite nanorods: soft-template synthesis, characterization and preliminary in vitro tests.

PubMed

Nguyen, Nga Kim; Leoni, Matteo; Maniglio, Devid; Migliaresi, Claudio

2013-07-01

Synthetic hydroxyapatite nanorods are excellent candidates for bone tissue engineering applications. In this study, hydroxyapatite nanorods resembling bone minerals were produced by using soft-template method with cetyltrimethylammonium bromide. Composite hydroxyapatite/poly(D, L)lactic acid films were prepared to evaluate the prepared hydroxyapatite nanorods in terms of cell affinity. Preliminary in vitro experiments showed that aspect ratio and film surface roughness play a vital role in controlling adhesion and proliferation of human osteoblast cell line MG 63. The hydroxyapatite nanorods with aspect ratios in the range of 5.94-7 were found to possess distinctive properties, with the corresponding hydroxyapatite/poly(D, L)lactic acid films promoting cellular confluence and a fast formation of collagen fibers as early as after 7 days of culture.

Managing the Mississippi River floodplain: Achieving ecological benefits requires more than hydrological connection to the river: Chapter

USGS Publications Warehouse

Schramm, Harold; Richardson, William B.; Knights, Brent C.

2015-01-01

Floodplains are vital to the structure and function of river-floodplain ecosystems. Among the many ecological services provided by floodplains are nutrient cycling and seasonal habitats for fish, including spawning, nursery, foraging and wintering habitats. Connections between the river channel and floodplain habitats are essential to realize these ecological services, but spatial and temporal aspects of the connection and contemporary geomorphology must also be considered in restoration efforts. This chapter synthesizes available information to compare floodplain function and needed management strategies in two extensive reaches (upper impounded and lower free-flowing) of the Mississippi River, USA. The upper impounded reach is the 523-km reach from about Minneapolis, Minnesota to Clinton, Iowa. This reach has been impounded and channelized for navigation. Mean annual water-level fluctuation ranges from 1 to 2 m in the navigation pools in this reach. Floodplain environmental conditions that affect nitrogen cycling and fish production vary seasonally and longitudinally within and among navigation pools. Significant issues affecting ecological services include sedimentation, constrained water level fluctuations, island erosion and seasonal hypoxia. The lower free-flowing reach, the 1570-km reach from the confluence of the Ohio and Mississippi rivers to the Gulf of Mexico, has no dams and average annual fluctuations of 7 m throughout most of the reach. Despite the substantial flood pulse, floodplain inundation is often brief and may not occur annually. Significant issues affecting floodplain ecological function are the short duration and thermal asynchrony of the flood pulse, sedimentation and loss of connection between the river channel and permanent/semi-permanent floodplain water bodies due to channel incision. Needs and strategies for floodplain enhancement to increase ecological services, particularly nitrogen cycling and fish production, differ along the longitudinal gradient of the Mississippi River and provide informative contrasts to guide floodplain management. Prediction of the effects of climate change on this system will be complicated by the magnitude of the watershed that encompasses 41 % of the continental USA and multiple climatic regions.

Modulating prime molecular expressions and in vitro wound healing rate in keratinocyte (HaCaT) population under characteristic honey dilutions.

PubMed

Chaudhary, Amrita; Bag, Swarnendu; Mandal, Mousumi; Krishna Karri, Sri Phani; Barui, Ananya; Rajput, Monika; Banerjee, Provas; Sheet, Debdoot; Chatterjee, Jyotirmoy

2015-05-26

In traditional medicines honey is known for healing efficacy and vividly used as "Anupan" in Ayurvedic medicines appreciating roles in dilutions. Validating efficacy of physico-chemically characterized honey in dilutions, studies on in vitro wound healing and attainment of cellular confluence epithelial cells including expressions of cardinal genes is crucial. To evaluate effects of characterized honey in varied dilutions on cellular viability, in vitro wound healing and modulation of prime epithelial gene expressions. Six Indian honey-samples from different sources were physico-chemically characterized and optimal one was explored in dilutions (v/v%) through in vitro studies on human epithelial (HaCaT) cells for viability, wound healing and expressions of genes p63, E-cadherin, β-catenin, GnT-III and GnT-V. Studied honey samples (i.e. A-F) depicted range of pH (2-4), water (12.48-23.95), electrical conductivity (2.57-14.34), carbohydrate (68.73-98.65), protein (.316-5.36) and antioxidant potential. Though sample A and F showed physico-chemical proximity, but overall bio-impact of the earlier was better, thus studied in 8-.1% (v/v) dilution range. Four dilutions (.01, .04, .1, .25 v/v%) augmented cellular viability but in vitro wound healing was fastest (p<.05) under .1%. Such efficacy was further documented for p63 up-regulation by immunocytochemistry and mRNA studies. The E-cadherin and β-catenin mRNA-expressions were also up-regulated and their proteins were predominantly cytoplasmic. E-cadherin up-regulation was corroborative with down-regulation and up-regulation of GnT-III and GnT-V respectively. Present study illustrated efficacy of particular honey dilution (.1%) with characteristic free radical scavenging activity in facilitating cell proliferation and attainment of confluence towards faster wound healing and modulation of cardinal epithelial genes (viz. p63, E-cadherin, β-catenin, Gnt-III and V). Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Accurate Morphology Preserving Segmentation of Overlapping Cells based on Active Contours

PubMed Central

Molnar, Csaba; Jermyn, Ian H.; Kato, Zoltan; Rahkama, Vesa; Östling, Päivi; Mikkonen, Piia; Pietiäinen, Vilja; Horvath, Peter

2016-01-01

The identification of fluorescently stained cell nuclei is the basis of cell detection, segmentation, and feature extraction in high content microscopy experiments. The nuclear morphology of single cells is also one of the essential indicators of phenotypic variation. However, the cells used in experiments can lose their contact inhibition, and can therefore pile up on top of each other, making the detection of single cells extremely challenging using current segmentation methods. The model we present here can detect cell nuclei and their morphology even in high-confluency cell cultures with many overlapping cell nuclei. We combine the “gas of near circles” active contour model, which favors circular shapes but allows slight variations around them, with a new data model. This captures a common property of many microscopic imaging techniques: the intensities from superposed nuclei are additive, so that two overlapping nuclei, for example, have a total intensity that is approximately double the intensity of a single nucleus. We demonstrate the power of our method on microscopic images of cells, comparing the results with those obtained from a widely used approach, and with manual image segmentations by experts. PMID:27561654

3D-printed PCL scaffolds for the cultivation of mesenchymal stem cells.

PubMed

Steffens, Daniela; Rezende, Rodrigo Alvarenga; Santi, Bruna; Pereira, Frederico David Alencar de Sena; Inforçatti Neto, Paulo; da Silva, Jorge Vicente Lopes; Pranke, Patricia

2016-04-06

Tissue engineering is a field which is currently under a great deal of investigation for the development and/or restoration of tissue and organs, through the combination of cell therapy with biomaterials. Rapid prototyping or additive manufacturing is a versatile technology which makes possible the fabrication of three dimensional (3D) structures from a wide range of materials with complex geometry and accuracy, such as scaffolds. The aim of this study has been to investigate the interaction between mesenchymal stem cells with poly (ε-caprolactone) (PCL) biomaterials used for obtaining scaffolds through additive manufacturing. Scanning electron microscopy, confocal microscopy and biological assays were performed to analyse the successful interaction between the cells and the biomaterials. As a result, the number of viable cells attached to the scaffolds was lower when compared to the control group; however, it was possible to observe cells in the scaffolds since day 1 of analysis, with regions of confluence after 21 days of seeding. To conclude, these biomaterials are interesting if used as medical artifacts, principally in tissue with prolonged regeneration time and which requires 3D supports with good mechanical properties.

Analysis of altimeter data jointly with seafloor electric data (vertically integrated velocity) and VCTD-yoyo data (detailed profiles of VCTD)

NASA Technical Reports Server (NTRS)

Tarits, Pascal D.; Menvielle, M.; Provost, C.; Filloux, J. H.

1991-01-01

We propose simultaneous analyses of the TOPEX/POSEIDON altimetry data, in situ data--mainly permanent seafloor electric recordings--and velocity, conductivity, temperature, density (VCTD)-yoyo data at several stations in areas of scientific interest. We are planning experiments in various areas of low and high energy levels. Several complementary and redundant methods will be used to characterize the ocean circulation and its short- and long-term variability. We shall emphasize long-term measurement using permanent stations. Our major initial objectives with the TOPEX/POSEIDON mission are the Confluence area in the Argentine Basin and the Circumpolar Antarctic Current. An early experiment was carried out in the Confluence zone in 1988 and 1990 (Confluence Principal Investigators, 1990) to prepare for an intensive phase later one. This intensive phase will include new types of instrumentation. Preliminary experiments will be carried out in the Mediterranean Sea (in 1991) and in the North Atlantic Ocean (in 1992, north of the Canary Islands) to test the new instrumentation.

The Arizona Wallow Wildfire: Monitoring It's Progress, Extreme Behavior and Long Range Smoke Transport from Multiple Satellite Platforms

NASA Astrophysics Data System (ADS)

Ruminski, M. G.; Fromm, M. D.; Ramirez, E.

2011-12-01

The Wallow fire in southeast Arizona was the largest wildfire in Arizona history, consuming over 500,000 acres. The fire began on May 29, 2011 and quickly grew to nearly 70,000 acres in size by June 4. This event exhibited anomalous behavioral characteristics as deep pyroconvection was observed for an unprecedented six consecutive days. The rapid spread and extreme pyroconvection occurred in response to a unique confluence of high biomass fuel availability in arid conditions. Strong winds in combination with low relative humidity and atmospheric instability acted to create an environment conducive to explosive fire growth. The resultant smoke from the blaze reached an altitude of nearly 15 km into the upper troposphere/lower stratosphere and eventually crossed the Atlantic reaching Europe. This presentation focuses on the detection and characterization of the Wallow fire from a satellite perspective. Geostationary and polar orbiting satellite platforms have captured various aspects of the fire and resulting smoke plumes. An animation of nearly 100 NOAA polar orbiting 4 micron channel images during the first 2 weeks of the fire illustrate its explosive growth while GOES visible channel animations display the density and coverage of the pall of smoke and the development of pyroconvection. True color MODIS imagery provides higher resolution views of the pyrocumulonimbus clouds at different stages in their evolution. CALIPSO, GOME2, and OMI data detail the vertical structure and composition of the plume as it drifts eastward and expands in coverage, eventually reaching Europe.

Assessing the concentration, speciation, and toxicity of dissolved metals during mixing of acid-mine drainage and ambient river water downstream of the Elizabeth Copper Mine, Vermont, USA

USGS Publications Warehouse

Balistrieri, L.S.; Seal, R.R.; Piatak, N.M.; Paul, B.

2007-01-01

The authors determine the composition of a river that is impacted by acid-mine drainage, evaluate dominant physical and geochemical processes controlling the composition, and assess dissolved metal speciation and toxicity using a combination of laboratory, field and modeling studies. Values of pH increase from 3.3 to 7.6 and the sum of dissolved base metal (Cd + Co + Cu + Ni + Pb + Zn) concentrations decreases from 6270 to 100 ??g/L in the dynamic mixing and reaction zone that is downstream of the river's confluence with acid-mine drainage. Mixing diagrams and PHREEQC calculations indicate that mixing and dilution affect the concentrations of all dissolved elements in the reach, and are the dominant processes controlling dissolved Ca, K, Li, Mn and SO4 concentrations. Additionally, dissolved Al and Fe concentrations decrease due to mineral precipitation (gibbsite, schwertmannite and ferrihydrite), whereas dissolved concentrations of Cd, Co, Cu, Ni, Pb and Zn decrease due to adsorption onto newly formed Fe precipitates. The uptake of dissolved metals by aquatic organisms is dependent on the aqueous speciation of the metals and kinetics of complexation reactions between metals, ligands and solid surfaces. Dissolved speciation of Cd, Cu, Ni and Zn in the mixing and reaction zone is assessed using the diffusive gradients in thin films (DGT) technique and results of speciation calculations using the Biotic Ligand Model (BLM). Data from open and restricted pore DGT units indicate that almost all dissolved metal species are inorganic and that aqueous labile or DGT available metal concentrations are generally equal to total dissolved concentrations in the mixing zone. Exceptions occur when labile metal concentrations are underestimated due to competition between H+ and metal ions for Chelex-100 binding sites in the DGT units at low pH values. Calculations using the BLM indicate that dissolved Cd and Zn species in the mixing and reaction zone are predominantly inorganic, which is consistent with the DGT results. Although the DGT method indicates that the majority of aqueous Cu species are inorganic, BLM calculations indicate that dissolved Cu is inorganic at pH 5.5. Integrated dissolved labile concentrations of Cd, Cu and Zn in the mixing and reaction zone are compared to calculated acute toxicity concentrations (LC50 values) for fathead minnows (Pimephales promelas) (Cd, Cu and Zn) and water fleas (Ceriodaphnia dubia) (Cd and Cu) using the BLM, and to national recommended water quality criteria [i.e., criteria maximum concentration (CMC) and criterion continuous concentration (CCC)]. Observed labile concentrations of Cd and Zn are below LC50 values and CMC for Cd, but above CCC and CMC for Zn at sites <30 m downstream of the confluence. In contrast, labile Cu concentrations exceed LC50 values for the organisms as well as CCC and CMC at sites <30 m downstream of the confluence. These results suggest that environmental conditions at sites closest to the confluence of the river and acid-mine drainage should not support healthy aquatic organisms. ?? 2007 Elsevier Ltd. All rights reserved.

The anti-estrogenic activity of sediments from agriculturally-intense watersheds: Assessment using in vivo and in vitro assays

PubMed Central

Sellin Jeffries, Marlo K.; Conoan, Nicholas H.; Cox, Marc B.; Sangster, Jodi L.; Balsiger, Heather A.; Bridges, Andrew A.; Cowman, Tim; Knight, Lindsey A.; Bartelt-Hunt, Shannon L.; Kolok, Alan S.

2015-01-01

The goal of the current study was to determine whether sediments from agriculturally-intense watersheds can act as a potential source of anti-estrogenic endocrine-disrupting compounds. The specific objectives of the current study were to determine 1) whether female fathead minnows (Pimephales promelas) experience alterations in endocrine function when exposed to sediments collected from agriculturally-intense watersheds and 2) if these sediments display anti-estrogenic activity in an in vitro assay. In addition, sediment samples were analyzed for the presence of steroid hormones and pesticides associated with local agricultural practices. To accomplish this, sediments and water were collected from three sites within two agriculturally-intense Nebraska watersheds (Bow Creek and the Elkhorn River). In 2009, minnows were exposed to sediment and/or water collected from the two Bow Creek sites (East Bow Creek and the Confluence) in the laboratory, while in 2010, minnows were exposed to sediment and/or water from East Bow Creek, the Confluence and the Elkhorn River. Following the 7-d exposure period, the hepatic mRNA expression of two-estrogen responsive genes, estrogen receptor α (ERα) and vitellogenin (Vtg) was determined. In 2009, females exposed to Confluence sediments, in the presence of laboratory water or Confluence water, experienced significant reductions in ERα expression relative to unexposed and Confluence water-exposed females. The defeminization of these females suggests the presence of a biologically-available anti-estrogenic compound in sediments collected from this site. In 2010, sediments were assessed for anti-estrogenic activity on days 0 and 7 of the exposure period using a four-hour yeast estrogen screen. Lipophilic extracts (LEs) of day 0 sediments collected from the Confluence and the Elkhorn River induced significant reductions in the estrogenic reporter activity of treated yeast cultures suggesting the presence of a lipophilic anti-estrogenic compound in these extracts. Chemical analysis revealed the presence of a variety of steroid hormones, including those associated with the production of beef cattle (ie: β-trenbolone, α-zearalanol and α-zearalenol), in sediments indicating that compounds utilized by local beef cattle operations are capable of entering nearby watersheds. Overall, the results of this study indicate that an environmentally-relevant anti-estrogenic compound is present in sediments from agriculturally-intense watersheds and that this compound is bioavailable to fish. Furthermore, the presence of steroid hormones in sediments from these watersheds provides evidence indicating that steroids are capable of sorbing to sediments. Clearly, sediments are capable of acting as a source of endocrine-disrupting compounds in the aquatic environment. PMID:21723217

Technical modification of the Balb/c 3T3 cell transformation assay: the use of serum-reduced medium to optimise the practicability of the protocol.

PubMed

Hayashi, Kumiko; Sasaki, Kiyoshi; Asada, Shin; Tsuchiya, Toshiyuki; Hayashi, Makoto; Yoshimura, Isao; Tanaka, Noriho; Umeda, Makoto

2008-12-01

The two-stage Balb/c 3T3 model of cell transformation can mimic the two-stage carcinogenicity bioassay, and has been recognised as a screening method for detecting potential tumour initiators and promoters. A technical modification to the original protocol (which involved the use of M10F medium, consisting of MEM plus 10% fetal bovine serum [FBS]) has been previously proposed, in order to increase its efficacy, namely: the introduction of enriched, serum-reduced medium (DF2F medium, comprising DMEM/F12 plus 2% FBS and other supplements). The aim of this study was to further modify the protocol, so as to attain higher practicability for the assay. The protocol was further optimised by: a) reducing the number of plates required, through the use of larger plates; b) reducing the cost of the assay by retaining the reduced serum concentration and by using 2microg/ml insulin, rather than the more-complex insulin-transferrin-ethanolamine-sodium selenite (ITES) supplement (i.e. DF2F2I medium); and c) extending the culture period from 24-25 days to 31-32 days, resulting in clearer foci (the number of medium changes did not increase, as less-frequent medium changes were performed during the extended culture period). Growth curve construction revealed that variations in the saturation densities of the parental Balb/c 3T3 cell line and its three transformed clones were highest when M10F medium was replaced with DF2F2I medium just before cells reached confluence. We applied this newly-optimised protocol to the assessment of: a) the tumour initiating activity of 3-methylcholanthrene (MCA), N-methyl-N'-nitro-N-nitrosoguanidine, mitomycin C, methylmethane sulphonate, CdCl(2) and phenacetin, combining a post-treatment of 100ng/ml 12-O-tetradecanoylphorbol-13-acetate at the promotion stage; and b) the tumour promoting activity of insulin, lithocholic acid, CdCl(2) and phenobarbital, with pre-treatment of 0.2microg/ml MCA at the initiation stage. In the present study, only phenobarbital was negative when tested by using the modified protocol. 2008 FRAME.

Production of CMAH Knockout Preimplantation Embryos Derived From Immortalized Porcine Cells Via TALE Nucleases.

PubMed

Moon, JoonHo; Lee, Choongil; Kim, Su Jin; Choi, Ji-Yei; Lee, Byeong Chun; Kim, Jin-Soo; Jang, Goo

2014-05-27

Although noncancerous immortalized cell lines have been developed by introducing genes into human and murine somatic cells, such cell lines have not been available in large domesticated animals like pigs. For immortalizing porcine cells, primary porcine fetal fibroblasts were isolated and cultured using the human telomerase reverse transcriptase (hTERT) gene. After selecting cells with neomycin for 2 weeks, outgrowing colonized cells were picked up and subcultured for expansion. Immortalized cells were cultured for more than 9 months without changing their doubling time (~24 hours) or their diameter (< 20 µm) while control cells became replicatively senescent during the same period. Even a single cell expanded to confluence in 100 mm dishes. Furthermore, to knockout the CMAH gene, designed plasmids encoding a transcription activator-like effector nuclease (TALENs) pairs were transfected into the immortalized cells. Each single colony was analyzed by the mutation-sensitive T7 endonuclease I assay, fluorescent PCR, and dideoxy sequencing to obtain three independent clonal populations of cells that contained biallelic modifications. One CMAH knockout clone was chosen and used for somatic cell nuclear transfer. Cloned embryos developed to the blastocyst stage. In conclusion, we demonstrated that immortalized porcine fibroblasts were successfully established using the human hTERT gene, and the TALENs enabled biallelic gene disruptions in these immortalized cells.

Production of CMAH Knockout Preimplantation Embryos Derived From Immortalized Porcine Cells Via TALE Nucleases

PubMed Central

Moon, JoonHo; Lee, Choongil; Kim, Su Jin; Choi, Ji-Yei; Lee, Byeong Chun; Kim, Jin-Soo; Jang, Goo

2014-01-01

Although noncancerous immortalized cell lines have been developed by introducing genes into human and murine somatic cells, such cell lines have not been available in large domesticated animals like pigs. For immortalizing porcine cells, primary porcine fetal fibroblasts were isolated and cultured using the human telomerase reverse transcriptase (hTERT) gene. After selecting cells with neomycin for 2 weeks, outgrowing colonized cells were picked up and subcultured for expansion. Immortalized cells were cultured for more than 9 months without changing their doubling time (~24 hours) or their diameter (< 20 µm) while control cells became replicatively senescent during the same period. Even a single cell expanded to confluence in 100 mm dishes. Furthermore, to knockout the CMAH gene, designed plasmids encoding a transcription activator-like effector nuclease (TALENs) pairs were transfected into the immortalized cells. Each single colony was analyzed by the mutation-sensitive T7 endonuclease I assay, fluorescent PCR, and dideoxy sequencing to obtain three independent clonal populations of cells that contained biallelic modifications. One CMAH knockout clone was chosen and used for somatic cell nuclear transfer. Cloned embryos developed to the blastocyst stage. In conclusion, we demonstrated that immortalized porcine fibroblasts were successfully established using the human hTERT gene, and the TALENs enabled biallelic gene disruptions in these immortalized cells. PMID:24866481

Isolation and characterization of mesenchymal progenitors derived from the bone marrow of goats native from northeastern Brazil.

PubMed

Silva Filho, Osmar Ferreira da; Argôlo Neto, Napoleão Martins; Carvalho, Maria Acelina Martins de; Carvalho, Yulla Klinger de; Diniz, Anaemilia das Neves; Moura, Laécio da Silva; Ambrósio, Carlos Eduardo; Monteiro, Janaína Munuera; Almeida, Hatawa Melo de; Miglino, Maria Angélica; Alves, Jacyara de Jesus Rosa Pereira; Macedo, Kássio Vieira; Rocha, Andressa Rego da; Feitosa, Matheus Levi Tajra; Alves, Flávio Ribeiro

2014-08-01

To characterize bone marrow progenitors cells grown in vitro, using native goats from northeastern Brazil as animal model. Ten northeastern Brazil native goats of both genders were used from the Piauí Federal University Agricultural Science Center's (UFPI) - Goat Farming Sector. Bone marrow aspirates where taken from the tibial ridge and seeded on culture plates for isolation, expansion and Flow Cytometry (expression markers - Oct-3/4, PCNA, Ck-Pan, Vimentina, Nanog). Progenitor cells showed colonies characterized by the presence of cell pellets with fibroblastoid morphology. Cell confluence was taken after 14 days culture and the non-adherent mononuclear cell progressive reduction. After the first passage, 94.36% cell viability was observed, starting from 4.6 x 106 cell/mL initially seeded. Cells that went through flow cytometry showed positive expression for Oct-3/4, PCNA, Ck-Pan, Vimentina, and Nanog. Bone marrow progenitor isolated of native goats from northeastern Brazil showed expression markers also seen in embryonic stem cells (Oct-3/4, Nanog), markers of cell proliferation (PCNA) and markers for mesenchymal cells (Vimentina and Ck-pan), which associated to morphological and culture growth features, suggest the existence of a mesenchymal stem cell (MSC) population in the goat bone marrow stromal cells studied.

Flood of April 1975 at Lansing, Michigan

USGS Publications Warehouse

Miller, John B.; Swallow, L.A.

1975-01-01

On April 18 between 5 p.m. and 12 p.m. an intense rainstorm fell in the Lansing area resulting in extensive flooding.  The Federal Disaster Assistance Administration estimated that 175 homes were damaged to at least half their value, 4,500 received some damage, with additional losses to schools, utilities, hospitals, and transportation systems.  Early estimates indicated that damages may be as high as $20 million.During the time of flooding the U.S. Geological Survey obtained aerial photography and streamflow data to document the disaster.  This report shows on photomosaic base maps the extent of flooding in the Lansing area.  Areas included are the lower reaches of the Red Cedar River and Sycamore Creek and the Grand River downstream from the confluence of the Red Cedar River.  Little flooding occurred on the Grand River upstream from the Red Cedar so, although aerial photography was obtained for that reach, photomosaics were not prepared.  Streamflow data collected at five gaging stations near Lansing are given.  Information on the magnitude of the flood should be useful in making decisions regarding use of flood plains in the area.  It is one of a series of reports on the April 1975 flood in the Lansing metropolitan area.

An Automated Method to Monitor Cell Migration.

NASA Astrophysics Data System (ADS)

Giaever, Ivar; Keese, Charles R.

2002-03-01

Electric cell-substrate impedance sensing (ECIS) has been developed as a non-invasive means to follow cell behavior in culture. In this method cells are cultured on small (250 micrometer diameter) gold film electrodes. The impedance of the electrode is measured by an AC current about 1 microampere. When challenged by biochemical or physical stimuli the cells will respond by changing their morphology and motion. These changes are reflected in the measured impedance values. In this study, the basic ECIS system was used in both a non-invasive and invasive mode to carry out an automated wound-healing assay for quantifying cell migration activity. BSC-1, MDCK, and NRK cell lines were grown to confluence in ECIS wells before data was collected. An AC current of approximately 1 milliampere at 40,000 Hz was applied for several seconds, killing the cells in contact with the ECIS electrode and dropping the impedance to that of a cell-free electrode. For the next few hours following this incursion, the neighboring cells migrate into the wounded area replacing the dead cells, and the electrodes return to impedance values of unwounded controls. Data shows that the time required for the completion of this activity is strongly dependent upon cell type, medium composition, and the type of protein adsorbed to the substrate.

[Abnormal expression of insulin-like growth factor-I receptor and inhibitory effect of its transcription intervention on nude mice xenograft tumor].

PubMed

Yao, M; Yan, X D; Cai, Y; Gu, J J; Yang, X L; Pan, L H; Wang, L; Yao, D F

2016-11-20

Objective: To investigate the expression of insulin-like growth factor-I receptor (IGF-IR) in liver cancer and the inhibitory effect of its transcription intervention on nude mice xenograft tumor. Methods: A total of 40 patients with primary liver cancer were enrolled, and 40 samples of cancer lesions, peri-cancerous tissues (with a distance of 2 cm to the margin of cancer lesion), or distal liver tissues (with a distance of 5 cm to the margin of cancer lesion), with a weight of 200 mg, were collected after surgery. Some of these samples were used for pathological examination, and the rest were stored at -85°C. A total of 18 BALB/c nude mice aged 4-6 weeks with a body weight of 18-20 g (9 male and 9 female mice) were randomly divided into control group, negative control group, and co-intervention group, with 6 mice in each group, and fed under specific pathogen-free conditions. The cell line was cultured in the dimethyl sulfoxide complete medium containing 10% fetal bovine serum in a CO 2 incubator at 37°C. When the cell confluence reached 90% after cell inoculation, shRNA was divided into co-intervention group, negative control group, and untreated control group and were transfected to hepatoma cells using PolyJetTM transfection reagent. Stable cell clones obtained by G418 screening and used for the in vivo study. Immunohistochemistry, Western blotting, and quantitative real-time PCR were used to analyze the expression of IGF-IR in the human hepatoma tissue and cell line. The IGF-IR shRNA eukaryotic expression plasmids were established and screened for the most effective sequence; they were transfected to PLC/PRF/5 hepatoma cells, and the CCK-8 assay was used to analyze the changes in cell proliferation. The stable cell line screened out by G418 was inoculated to establish the subcutaneous xenograft tumor in nude mice. The tumor growth curve was plotted and histological examination was performed. Graphpad Prism 5.0 and SPSS 18.0 were used for plotting and data analysis; the variance test and Q test were used for comparison of means between multiple samples, the t-test was used for comparison of means between any two samples, the chi-square test or Fisher's exact test was used for comparison of rates between samples, and a rank correlation analysis was performed for expression intensity. Results: The liver cancer group had a significantly higher positive rate of IGF-IR than the peri-cancerous group and distal tissue group (82.5% vs 42.5%/10%, χ 2 = 13.653 and 42.29, both P < 0.01), as well as significantly higher expression intensity than these two groups ( Z = 4.771 and 6.579, both P < 0.01). IGF-IR was not significantly expressed in the L02 cell line and was strongly expressed in the PLC/PRF/5 hepatoma cells, and the expression intensity of IGF-IR in the PLC/PRF/5 hepatoma cells was 4 and 5 times that in Bel-7404 cells and HepG2 cells, respectively. After the PLC/PRF/5 hepatoma cells were transfected with shRNA4 with the best co-intervention effect, the mean inhibition rate of tumor cell growth reached 63.9% at 72 hours, and the mean inhibition rate of IGF-IR transcription reached 59.6%. Tumor cells were arrested in G1 phase, and there was a significant increase in apoptosis rate. As for the subcutaneous hepatoma xenograft in nude mice, the intervention group had significantly slower tumor growth than the blank control group and negative control group (143±24 mm3 vs 372±46 mm3/350±50 mm3, t = 10.776 and 9.142, both P < 0.01); the intervention group had significantly downregulated IGF-IR expression, which was significantly lower than that in the blank control group and negative control group ( t = 11.184 and 9.450, both P < 0.01). Conclusion: Intervention of IGF-IR transcription can effectively inhibit the growth of xenograft tumor in nude mice, suggesting that IGF-IR gene might become a new potential target for the treatment of liver cancer.

River conferences under temperate valley glaciers

NASA Astrophysics Data System (ADS)

Lane, Stuart; Egli, Pascal; Irving, James

2017-04-01

Both geophysical measurements (ground penetrating radar) and hydrological inference has shown that subglacial drainage networks are dendritic and that means that they must have confluences. In general, there are very few studies of rivers under glaciers and almost no consideration at all of confluences, despite the fact that they could be a critical parameter in understanding coupling at the ice-sediment bed interface. Subglacial channels, normally known as conduits, are typically associated with the combined effect of hydraulic pressure driven ice melt (which opens them) and ice overburden pressure (which closes them). Inferences from dye break out curves shows that has the efficiency of ice melt increases progressively during the summer ablation season, melt rates closure rates and a channelized system becomes progressively more effective. Most recently, measurements at the Upper Arolla Glacier show that the effects of this growing efficiency is an evolution in the subglacial hydrological system towards higher peak flows and lower base flows later in the melt season. This increases the probability that late in the melt season, sediment transport becomes discontinuous, with overnight deposition and daytime erosion. This would in turn produce the rapid reductions in sediment transport capacity overnight needed to deposit sediment and to block conduits, increase basal water pressure and explain the hydraulic jacking observed in snout marginal zones at a time when it should not be expected. The question that follows is what effects do confluences have on this process? The geometry of subglacial channels is such that when they join they lead to rapid changes in hydraulic geometry. Crucially, these are likely to have a non-linear impact upon sediment transport capacity, which should reduce disproportionally in the conduits downstream of the junction. Thus, it is possible that confluence zones under glaciers become sites of very rapid sediment accumulation and blockage overnight. In this paper, we present some one-dimensional coupled hydraulic sediment transport modelling to show this process. It suggests that the dendritic form of the subglacial drainage network is the primary reason why sediment blockage occurs and suggests the need for a more in-depth assessment of how sediment moves through confluences under glaciers.

Hydraulic and Geomorphic Effects of Large Woody Debris Additions in the Narraguagus Watershed, Coastal Maine

NASA Astrophysics Data System (ADS)

Johnson, E.; Snyder, N. P.

2009-12-01

Maine coastal rivers host the last remaining runs of endangered anadromous Atlantic salmon in the United States, whose populations have declined from ~500,000 returning adults in the 1880s to only ~1000 in 2000. Restoration projects focus on these rivers, and recent efforts involve adding large woody debris (LWD) to small tributaries to improve salmon rearing habitat. LWD changes the hydraulics and geomorphology of small streams by acting as a barrier to flow and resulting in decreased velocity zones, scour pools, and sediment storage and sorting. We quantify the effects of LWD additions in early August 2008 on hydraulics and substrate in Baker Brook, a west-flowing tributary of the Narraguagus River. Hydraulically, we focus on the treatment reach nearest the confluence with the Narraguagus River (Baker1). We study substrate in this and the upstream treatment location (Baker3). Both study locations are divided into two reaches, treatment (received cut wood) and control (did not receive cut wood). The reaches are further divided into 50 m sub-reaches based on gradient (1-2%). We seek to answer two research questions: (1) how much does mean velocity through the study sub-reaches change as a result of additions; and (2) does sediment storage and sorting result from the LWD additions? We measured reach-average velocities (Ureach) in Baker1 using the salt dilution method in May, July, August and October 2008 and May 2009. We use rating curves to compare the post-treatment to the pre-treatment Ureach-stage relationship. No reach-average change was quantified during the low-to-moderate flows we observed. We found a localized change in cross-sectionally averaged velocity (U⊥) measured with a flow meter in one sub-reach, but this is because an added tree intersected the cross-section where measurements were recorded. We quantified pre- and post-treatment sub-reach substrate median grain size (D50) with intensive clast counts in July 2008 and May 2009. Two of the three treatment sub-reaches exhibited significant fining (D50 decreased by 37-54%) between the surveys, and the other changed less than measurement uncertainty. Two of the three control sub-reaches remained unchanged, and one fined significantly (-42%). In summary, we find that LWD additions in Baker Brook did not affect reach-scale hydraulics during the flows we observed, but did influence bed-grain size during the 10-month study interval, underscoring the importance of floods on channel change.

Iatrogenic bile duct injury with loss of confluence.

PubMed

Mercado, Miguel-Angel; Vilatoba, Mario; Contreras, Alan; Leal-Leyte, Pilar; Cervantes-Alvarez, Eduardo; Arriola, Juan-Carlos; Gonzalez, Bruno-Adonai

2015-10-27

To describe our experience concerning the surgical treatment of Strasberg E-4 (Bismuth IV) bile duct injuries. In an 18-year period, among 603 patients referred to our hospital for surgical treatment of complex bile duct injuries, 53 presented involvement of the hilar confluence classified as Strasberg E4 injuries. Imagenological studies, mainly magnetic resonance imaging showed a loss of confluence. The files of these patients were analyzed and general data were recorded, including type of operation and postoperative outcome with emphasis on postoperative cholangitis, liver function test and quality of life. The mean time of follow-up was of 55.9 ± 52.9 mo (median = 38.5, minimum = 2, maximum = 181.2). All other patients with Strasberg A, B, C, D, E1, E2, E3, or E5 biliary injuries were excluded from this study. Patients were divided in three groups: G1 (n = 21): Construction of neoconfluence + Roux-en-Y hepatojejunostomy. G2 (n = 26): Roux-en-Y portoenterostomy. G3 (n = 6): Double (right and left) Roux-en-Y hepatojejunostomy. Cholangitis was recorded in two patients in group 1, in 14 patients in group 2, and in one patient in group 3. All of them required transhepatic instrumentation of the anastomosis and six patients needed live transplantation. Loss of confluence represents a surgical challenge. There are several treatment options at different stages. Roux-en-Y bilioenteric anastomosis (neoconfluence, double-barrel anastomosis, portoenterostomy) is the treatment of choice, and when it is technically possible, building of a neoconfluence has better outcomes. When liver cirrhosis is shown, liver transplantation is the best choice.

Geological, hydrological, and biological issues related to the proposed development of a park at the confluence of the Los Angeles River and the Arroyo Seco, Los Angeles County, California

USGS Publications Warehouse

Land, Michael; Trenham, Peter C.; Ponti, Daniel J.; Reichard, Eric G.; Tinsley, John C.; Warrick, Jonathan A.; Meyer, Robert W.

2005-01-01

A new park is being considered for the confluence of the Los Angeles River and the Arroyo Seco in Los Angeles County, California. Components of the park development may include creation of a temporary lake on the Los Angeles River, removal of channel lining along part of the Arroyo Seco, restoration of native plants, creation of walking paths, and building of facilities such as a boat ramp and a visitor center. This report, prepared in cooperation with the Mountains Recreation and Conservancy Authority, delineates the geological, hydrological, and biological issues that may have an impact on the park development or result from development at the confluence, and identifies a set a tasks to help address these science issues. Geologic issues of concern relate to surface faulting, earthquake ground motions, liquefaction, landsliding, and induced seismicity. Hydrologic issues of concern relate to the hydraulics and water quality of both surface water and ground water. Biological issues of concern include colonization-extinction dynamics, wildlife corridors, wildlife reintroduction, non-native species, ecotoxicology, and restoration of local habitat and ecology. Potential tasks include (1) basic data collection and follow-up monitoring, and (2) statistical and probabilistic analyses and simulation modeling of the seismic, hydraulic, and ecological processes that may have the greatest impact on the park. The science issues and associated tasks delineated for the proposed confluence park will also have transfer value for river restoration in other urban settings.

Culture of prostate epithelial cells of the rhesus monkey on extracellular matrix substrate: influence of steroids and insulin-like growth factors.

PubMed

Udayakumar, T S; Jeyaraj, D A; Rajalakshmi, M; Sharma, R S

1999-09-01

Rhesus monkey prostate epithelial cells from the cranial lobe were isolated and cultured in flasks coated either with collagen IV or laminin. The effects of stromal cell medium, androgens and growth factors on cell number, thymidine incorporation and secretory activity were assessed. The results indicate that dihydrotestosterone (DHT) and androstenedione have stimulatory influences on cell proliferation and secretion in coated flasks. DHT was more effective in increasing cell number but the induction of secretory activity was similar with both steroids. The combination of IGF-I and -II resulted in inducing better cell proliferation and secretory activity than the individual IGFs but, of the two IGFs, IGF-I was more effective than IGF-II. DHT with IGFs was more potent in inducing proliferation, differentiation and secretion than androstenedione. Even in the absence of steroids or growth factors, colony formation and confluence occurred in coated flasks but cell differentiation and secretion only to a limited extent. In conclusion, we were able to establish an in vitro primary culture of prostate epithelial cells from rhesus monkey using extracellular matrix proteins, steroids and growth factors as additional supplements. This culture system may be useful to study prostate cell physiology and to identify drugs that can inhibit cell proliferation.

Involvement of CTGF, a hypertrophic chondrocyte-specific gene product, in tumor angiogenesis.

PubMed

Shimo, T; Nakanishi, T; Nishida, T; Asano, M; Sasaki, A; Kanyama, M; Kuboki, T; Matsumura, T; Takigawa, M

2001-01-01

Connective tissue growth factor (CTGF) is a potent secreted signaling factor which functions in multiple stages of angiogenesis. In the present study, we examined the role of CTGF in tumor angiogenesis and made the following observations: (1) Histological analysis of human breast cancer (MDA231) cell and human fibrosarcoma (HT1080) cell xenografts in BALB/c nude mice showed a high level of neovascularization. Human squamous cell carcinoma (A431) xenografts induced only a low level of neovascularization. (2) CTGF mRNA was strongly expressed in MDA231 and in HT1080 cells in vivo and in vitro, but not in A431 cells. (3) CTGF protein was markedly produced in MDA231 cells and HT1080 cells and secreted into culture medium, and its production was greater during phases of growth rather than confluency. (4) Production of CTGF in bovine aorta endothelial cells was induced by CTGF, VEGF, bFGF and TGF-beta. (5) Neovascularization induced by HT1080 cells or MDA231 cells on chicken chorioallantoic membrane was suppressed in the presence of neutralizing CTGF-specific polyclonal antibody. These results suggest that CTGF regulates progression in tumor angiogenesis and the release or secretion of CTGF from tumor cells is essential for the angiogenesis. Copyright 2001 S. Karger AG, Basel

Flood-inundation maps for the Green River in Colrain, Leyden, and Greenfield, Massachusetts, from U.S. Geological Survey streamgage 01170100 Green River near Colrain to the confluence with the Deerfield River

USGS Publications Warehouse

Flynn, Robert H.; Bent, Gardner C.; Lombard, Pamela J.

2016-09-02

The U.S. Geological Survey developed flood elevations in cooperation with the Federal Emergency Management Agency for a 14.3-mile reach of the Green River in Colrain, Leyden, and Greenfield, Massachusetts, to assist landowners and emergency management workers to prepare for and recover from floods. The river reach extends from the U.S. Geological Survey Green River near Colrain, MA (01170100) streamgage downstream to the confluence with the Deerfield River. A series of seven digital flood inundation maps were developed for the upper 4.4 miles of the river reach downstream from the stream. Flood discharges corresponding to the 50-, 10-, 1-, and 0.2-percent annual exceedance probabilities were computed for the reach from updated flood-frequency analyses. These peak flows and the flood flows associated with the stages of 10.2, 12.4, and 14.4 feet (ft) at the Green River streamgage were routed through a one-dimensional step-backwater hydraulic model to obtain the corresponding peak water-surface elevations and to place the Tropical Storm Irene flood of August 28, 2011 (stage 13.97 ft), into historical context. The hydraulic model was calibrated by using the current (2015) stage-discharge relation at the U.S. Geological Survey Green River near Colrain, MA (01170100) streamgage and from documented high-water marks from the Tropical Storm Irene flood, which had a flow higher than a 0.2-percent annual exceedance probability flood discharge.The hydraulic model was used to compute water-surface profiles for flood stages referenced to the streamgage and ranging from the 50-percent annual exceedance probability (bankfull flow) at 7.6 ft (439.8 ft above the North American Vertical Datum of 1988 [NAVD 88]) to 14.4 ft (446.7 ft NAVD 88), which exceeds the maximum recorded water level of 13.97 ft (Tropical Storm Irene) at the streamgage. The mapped stages of 7.6 to 14.4 ft were selected to match the stages for bankfull; the 50-, 10-, 1-, and 0.2-percent annual exceedance probabilities; incremental stages of 10.2 and 12.4 ft; and the maximum stage of the stage-discharge rating curve. The simulated water-surface profiles were combined with a geographic information system digital elevation model derived from light detection and ranging (lidar) data having a 0.5-ft vertical accuracy to create a set of flood-inundation maps.The availability of the flood-inundation maps, combined with information regarding near real-time stage from U.S. Geological Survey Green River near Colrain, MA (01170100) streamgage, can provide emergency management personnel and residents with information that is critical for flood response activities, such as evacuations and road closures, and postflood recovery efforts. The flood-inundation maps are nonregulatory but provide Federal, State, and local agencies and the public with estimates of the potential extent of flooding during selected peak-flow events.

Bedload transport in a river confluence

NASA Astrophysics Data System (ADS)

Martín-Vide, J. P.; Plana-Casado, A.; Sambola, A.; Capapé, S.

2015-12-01

The confluence of the regulated Toltén River and its tributary the unregulated Allipén (south of Chile) has proved dynamic in the last decade. Daily bedload measurements with a Helley-Smith sampler, bed surveys, and grain-size distributions of the two rivers are obtained from a field campaign that lasts 3 months in high-flow season. The goals are to quantify total bedload and to understand the balance between tributary and main river and the bedload distribution in space and texture. The bedload transport varies 200-fold, with a maximum of 5000 t/day. The discharge varies five-fold, with a maximum of 900 m3/s. Two-thirds of the total bedload volume are transported through the deeper area of the cross section and gravel is predominant (64%). Average bedload volumes in the confluence seem unbalanced in favour of the tributary. Main river bedload transport is predominantly at below-capacity conditions, while the tributary bedload transport is at-capacity conditions. This is deemed the main reason of inaccuracy of the bedload predictors. The roles of entrainment into suspension, helical flow, partial transport, and mobile armour are discussed.

A Comparison of Real-Time and Endpoint Cell Viability Assays for Improved Synthetic Lethal Drug Validation.

PubMed

Single, Andrew; Beetham, Henry; Telford, Bryony J; Guilford, Parry; Chen, Augustine

2015-12-01

Cell viability assays fulfill a central role in drug discovery studies. It is therefore important to understand the advantages and disadvantages of the wide variety of available assay methodologies. In this study, we compared the performance of three endpoint assays (resazurin reduction, CellTiter-Glo, and nuclei enumeration) and two real-time systems (IncuCyte and xCELLigence). Of the endpoint approaches, both the resazurin reduction and CellTiter-Glo assays showed higher cell viabilities when compared directly to stained nuclei counts. The IncuCyte and xCELLigence real-time systems were comparable, and both were particularly effective at tracking the effects of drug treatment on cell proliferation at sub-confluent growth. However, the real-time systems failed to evaluate contrasting cell densities between drug-treated and control-treated cells at full growth confluency. Here, we showed that using real-time systems in combination with endpoint assays alleviates the disadvantages posed by each approach alone, providing a more effective means to evaluate drug toxicity in monolayer cell cultures. Such approaches were shown to be effective in elucidating the toxicity of synthetic lethal drugs in an isogenic pair of MCF10A breast cell lines. © 2015 Society for Laboratory Automation and Screening.

Onyx embolization of anterior condylar confluence dural arteriovenous fistula

PubMed Central

Takemoto, Koichiro; Tateshima, Satoshi; Rastogi, Sachin; Gonzalez, Nestor; Jahan, Reza; Duckwiler, Gary; Vinuela, Fernando

2013-01-01

The anterior condylar confluence (ACC) is a small complex venous structure located medial to the jugular vein and adjacent to the hypoglossal canal. To our knowledge, this is the first report of transvenous Onyx embolization for ACC dural arteriovenous fistula (DAVF). Three patients with ACC DAVF were treated using the Onyx liquid embolic agent with or without detachable coils. Complete angiographic obliteration of the fistulas was achieved in all cases without permanent lower cranial neuropathy. This report suggests that the controlled penetration of Onyx is advantageous in order to obliterate ACC DAVFs with a small amount of embolic material. PMID:23459160

Microscale diffusion measurements and simulation of a scaffold with a permeable strut.

PubMed

Lee, Seung Youl; Lee, Byung Ryong; Lee, Jongwan; Kim, Seongjun; Kim, Jung Kyung; Jeong, Young Hun; Jin, Songwan

2013-10-10

Electrospun nanofibrous structures provide good performance to scaffolds in tissue engineering. We measured the local diffusion coefficients of 3-kDa FITC-dextran in line patterns of electrospun nanofibrous structures fabricated by the direct-write electrospinning (DWES) technique using the fluorescence recovery after photobleaching (FRAP) method. No significant differences were detected between DWES line patterns fabricated with polymer supplied at flow rates of 0.1 and 0.5 mL/h. The oxygen diffusion coefficients of samples were estimated to be ~92%-94% of the oxygen diffusion coefficient in water based on the measured diffusion coefficient of 3-kDa FITC-dextran. We also simulated cell growth and distribution within spatially patterned scaffolds with struts consisting of either oxygen-permeable or non-permeable material. The permeable strut scaffolds exhibited enhanced cell growth. Saturated depths at which cells could grow to confluence were 15% deeper for the permeable strut scaffolds than for the non-permeable strut scaffold.

Tangeritin inhibits adipogenesis by down-regulating C/EBPα, C/EBPβ, and PPARγ expression in 3T3-L1 fat cells.

PubMed

He, Y F; Liu, F Y; Zhang, W X

2015-10-29

The treatment of obese patients is a topic investigated by an increasing number of researchers. This study aimed to elucidate the possible inhibitory effect of tangeritin on the development and function of fat cells. 3T3-L1 fat cells were grown to confluence and subjected to different concentrations of tangeritin. The most effective tangeritin inhibition concentration was determined by the MTT assay. The treated cells were subjected to real-time reverse transcriptase PCR and western blot analysis, to detect changes in the CCAAT/enhancer binding protein (C/EBP)α, C/EBPβ, and peroxisome proliferator activated receptor (PPAR)γ expression levels. The MTT assay revealed that the fat cell growth was inhibited at a 20 ng/mL concentration of tangeritin. The results of real-time PCR revealed a significant decrease in the expression of C/EBPα, C/EBPβ, and PPARγ mRNA, following the treatment with tangeritin. Western blot analysis also presented similar results at a protein level. Therefore, we concluded that tangeritin inhibits adipogenesis via the down-regulation of C/EBPα, C/EBPβ, and PPARγ mRNA and protein expression in 3T3-L1 cells.

Differential Properties of Human ALP+ Periodontal Ligament Stem Cells vs Their ALP- Counterparts

PubMed Central

Tran, Quynh T; El-Ayachi, Ikbale; Bhatti, Fazal-Ur-Rehman; Bahabri, Rayan; Al-Habib, Mey; Huang, George TJ

2015-01-01

Characterizing subpopulations of stem cells is important to understand stem cell properties. Tissue-nonspecific alkaline phosphatase (ALP) is associated with mineral tissue forming cells as well as stem cells. Information regarding ALP subpopulation of human periodontal ligament stem cells (hPDLSCs) is limited. In the present study, we examined ALP+ and ALP− hPDLSC subpopulations, their surface markers STRO-1 and CD146, and the expression of stemness genes at various cell passages. We found that ALP+ subpopulation had higher levels of STRO-1 (30.6 ± 5.6%) and CD146 (90.4 ± 3.3%) compared to ALP− (STRO-1: 0.5 ± 0.1%; CD146: 75.3 ± 7.2%). ALP+ cells expressed significantly higher levels of stemness associated genes, NANOG, OCT4 and SOX than ALP− cells at low cell passages of 2-3 (p<0.05). ALP+ and ALP− cells had similar osteogenic, chondrogenic and neurogenic potential while ALP−, not ALP+ cells, lacked adipogenic potential. Upon continuous culturing and passaging, ALP+ continued to express higher stemness genes and STRO-1 and CD146 than ALP− cells at ≥passage 19. Under conditions (over-confluence and vitamin C treatment) when ALP+ subpopulation was increased, the stemness gene levels of ALP+ was no longer significantly higher than those in ALP− cells. In conclusion, ALP+ hPDLSCs possess differential properties from their ALP− counterparts. PMID:26807329

Polarity and transport properties of rabbit kidney proximal tubule cells on collagen IV-coated porous membranes.

PubMed

Genestie, I; Morin, J P; Vannier, B; Lorenzon, G

1995-07-01

A high degree of functional polarity has been obtained in primary cultures of rabbit kidney proximal tubule cells grown on collagen IV-coated porous membranes. Tight confluency was attained 6 days after seeding and maintained for at least 6 more days, as shown by analysis of paracellular inulin diffusion. From day 6 onward, L-lactate, ammonia, and D-glucose concentration gradient and a pH difference of approximately 1 unit developed between the two nutrient medium compartments. Confluent monolayers expressed organic ion transport properties higher than those formerly reported for other cell models. Transcellular transport of 20 microM tetraethylammonium was directed from basal to apical compartment and was specifically inhibited by mepiperphenidol (1 mM). Unidirectional transport of 2.4 microM p-aminohippurate also occurred from basal to apical compartment, was saturable, and specifically inhibited by probenecid (1 mM). These results suggest that rabbit kidney proximal tubule cells, cultured under the experimental conditions described here, may be a useful model for the in vitro study of highly polarized renal transport processes.

Iatrogenic bile duct injury with loss of confluence

PubMed Central

Mercado, Miguel-Angel; Vilatoba, Mario; Contreras, Alan; Leal-Leyte, Pilar; Cervantes-Alvarez, Eduardo; Arriola, Juan-Carlos; Gonzalez, Bruno-Adonai

2015-01-01

AIM: To describe our experience concerning the surgical treatment of Strasberg E-4 (Bismuth IV) bile duct injuries. METHODS: In an 18-year period, among 603 patients referred to our hospital for surgical treatment of complex bile duct injuries, 53 presented involvement of the hilar confluence classified as Strasberg E4 injuries. Imagenological studies, mainly magnetic resonance imaging showed a loss of confluence. The files of these patients were analyzed and general data were recorded, including type of operation and postoperative outcome with emphasis on postoperative cholangitis, liver function test and quality of life. The mean time of follow-up was of 55.9 ± 52.9 mo (median = 38.5, minimum = 2, maximum = 181.2). All other patients with Strasberg A, B, C, D, E1, E2, E3, or E5 biliary injuries were excluded from this study. RESULTS: Patients were divided in three groups: G1 (n = 21): Construction of neoconfluence + Roux-en-Y hepatojejunostomy. G2 (n = 26): Roux-en-Y portoenterostomy. G3 (n = 6): Double (right and left) Roux-en-Y hepatojejunostomy. Cholangitis was recorded in two patients in group 1, in 14 patients in group 2, and in one patient in group 3. All of them required transhepatic instrumentation of the anastomosis and six patients needed live transplantation. CONCLUSION: Loss of confluence represents a surgical challenge. There are several treatment options at different stages. Roux-en-Y bilioenteric anastomosis (neoconfluence, double-barrel anastomosis, portoenterostomy) is the treatment of choice, and when it is technically possible, building of a neoconfluence has better outcomes. When liver cirrhosis is shown, liver transplantation is the best choice. PMID:26527428

Ethics in cryptomarket research.

PubMed

Martin, James; Christin, Nicolas

2016-09-01

The recent proliferation of cryptomarkets and the associated emergence of a sub-field of research on the anonymous web have outpaced the development of an ethical consensus regarding research methods and dissemination amongst scholars working in this unique online space. The peculiar characteristics of cryptomarket research, which often involves encryption, illegal activity, large-scale data collection, and geographical separation from research participants, challenge conventional ethical frameworks. A further complicating factor for reaching ethical consensus is the confluence of scholars drawn from a variety of academic disciplines, each with their own particular norms, practices and perspectives. This paper is intended to stimulate awareness and debate, and to prompt further reflection amongst scholars studying these fascinating online phenomena. The paper explores tensions and addresses some of the more prominent and pressing ethical questions, including public vs. private online spaces, anonymity, data sharing and ownership, risks and threats to research subjects and researchers. Also discussed is how best to balance the potential harms of cryptomarket research against benefits to the public. Copyright © 2016 Elsevier B.V. All rights reserved.

Flood of September 3, 1972, in Hillsboro, New Mexico

USGS Publications Warehouse

Waite, Loyd A.

1973-01-01

Four deaths, two persons seriously injured, and property damage estimated at $846,500 resulted from a flood that struck the village of Hillsboro, N. Mex. during the early morning hours of September 3, 1972. Hillsboro is situated at the confluence of Percha Creek and its main tributary, North Percha Creek. Heavy rainfall in the drainage basin upstream from Hillsboro occurred during the night of September 2-3. Peak discharge computed by slope-area measurements indicated that Percha Creek carried a maximum flow of 12,200 cubic feet per second (346 cubic meters per second) and that North Percha Creek carried a flow of 20,900 cubic feet per second (592 cubic meters per second) at points just upstream from Hillsboro. Flood-control facilities in Hillsboro consisting of levees on the south bank of Percha Creek failed and flood waters reached depths of 6 feet (1.8 meters) in Hillsboro. Hillsboro was declared a disaster area by Governor Bruce King and by President Richard Nixon.

Immersed Boundary Models for Quantifying Flow-Induced Mechanical Stimuli on Stem Cells Seeded on 3D Scaffolds in Perfusion Bioreactors.

PubMed

Guyot, Yann; Smeets, Bart; Odenthal, Tim; Subramani, Ramesh; Luyten, Frank P; Ramon, Herman; Papantoniou, Ioannis; Geris, Liesbet

2016-09-01

Perfusion bioreactors regulate flow conditions in order to provide cells with oxygen, nutrients and flow-associated mechanical stimuli. Locally, these flow conditions can vary depending on the scaffold geometry, cellular confluency and amount of extra cellular matrix deposition. In this study, a novel application of the immersed boundary method was introduced in order to represent a detailed deformable cell attached to a 3D scaffold inside a perfusion bioreactor and exposed to microscopic flow. The immersed boundary model permits the prediction of mechanical effects of the local flow conditions on the cell. Incorporating stiffness values measured with atomic force microscopy and micro-flow boundary conditions obtained from computational fluid dynamics simulations on the entire scaffold, we compared cell deformation, cortical tension, normal and shear pressure between different cell shapes and locations. We observed a large effect of the precise cell location on the local shear stress and we predicted flow-induced cortical tensions in the order of 5 pN/μm, at the lower end of the range reported in literature. The proposed method provides an interesting tool to study perfusion bioreactors processes down to the level of the individual cell's micro-environment, which can further aid in the achievement of robust bioprocess control for regenerative medicine applications.

Morphology of primary human venous endothelial cell cultures before and after culture medium exchange.

PubMed

Krüger-Genge, A; Fuhrmann, R; Jung, F; Franke, R P

2015-01-01

The evaluation of the interaction of human, venous endothelial cells (HUVEC) with body foreign materials on the cellular level cannot be performed in vivo, but is investigated in vitro under standard culture conditions. To maintain the vitality, proliferation and morphology of HUVEC seeded on body foreign substrates over days, the cell culture medium is usually exchanged every second day. It is well known, that alterations in the microenvironment of cells bear the risk of influencing cell morphology and function. In the current study the influence of cell culture medium exchange on HUVEC cytoskeletal microfilament structure and function was investigated. HUVEC in the third passage were seeded on extracellular matrix (ECM) - which was secreted from bovine corneal endothelial cells on glass- until functional confluence was reached. The experiment started 11 days after HUVEC seeding with an exchange of the cell culture medium followed by a staining of the actin microfilaments with phalloidin-rhodamin 1.5 and 5 minutes after medium exchange. The microfilaments were documented by use of an Olympus microscope (IMT-2) equipped with a UV lamp and online connected to a TV chain (Sony XC 50 ST/monochrome) implying an OPTIMAS - Image analysis system. Prostacyclin was analysed in the cell culture supernatant. 1.5 min after culture medium exchange in the functionally confluent cultures a slight disturbance of the actin microfilament structure with a broadening of the marginal filament band, a partial disconnection of cell-cell contacts and the appearance of intercellular fenestrations were observed. 5 minutes after medium exchange a redevelopment of the slightly disturbed microfilament structure with a condensation and narrowing of the marginal filament band was seen. 12 h later a further consolidation of the microfilament structure occurred. In addition, a perturbation of the cultured HUVEC occurred after cell culture medium exchange. The prostacyclin concentration in the supernatant increased significantly after 1.5 min to 466 ± 543 pg·mL-1 (p <  0.001) and after 5 min to 408 ± 458 pg·mL-1 (p <  0.001), while in control cells the prostacyclin concentration did not change remaining in the range of 50 ± 48.9 pg·mL-1. This study revealed that the exchange of the cell culture medium led to a rapid disturbance of the HUVEC with stress fiber formation, disconnection of cell-cell contacts and an altered prostacyclin secretion, which had regressed nearly completely after 12 hours. Therefore, the evaluation of HUVEC on body foreign materials should be performed not earlier than 12 hours after cell culture medium exchange to avoid a misinterpretation of the endothelial cell morphological state. This procedure minimizes the risk of a misinterpretation of the endothelial cell morphology - caused by the culture medium exchange and not by the interaction between biomaterials and HUVEC.

Gene expression of stem cells at different stages of ontological human development.

PubMed

Allegra, Adolfo; Altomare, Roberta; Curcio, Patrizia; Santoro, Alessandra; Lo Monte, Attilio I; Mazzola, Sergio; Marino, Angelo

2013-10-01

To compare multipotent mesenchymal stem cells (MSCs) obtained from chorionic villi (CV), amniotic fluid (AF) and placenta, with regard to their phenotype and gene expression, in order to understand if MSCs derived from different extra-embryonic tissues, at different stages of human ontological development, present distinct stemness characteristics. MSCs obtained from 30 samples of CV, 30 of AF and 10 placentas (obtained from elective caesarean sections) were compared. MSCs at second confluence cultures were characterized by immunophenotypic analysis with flow cytometry using FACS CANTO II. The expression of the genes Oct-4 (Octamer-binding transcription factor 4, also known as POU5F1), Sox-2 (SRY box-containing factor 2), Nanog, Rex-1 (Zfp-42) and Pax-6 (Paired Box Protein-6), was analyzed. Real-time quantitative PCR was performed by ABI Prism 7700, after RNA isolation and retro-transcription in cDNA. Statistical analysis was performed using non-parametric test Kruskal-Wallis (XLSTAT 2011) and confirmed by REST software, to estimate fold changes between samples. Each gene was defined differentially expressed if p-value was <0.05. Cells from all samples were negative for haematopoietic antigens CD45, CD34, CD117 and CD33 and positive for the typical MSCs antigens CD13, CD73 and CD90. Nevertheless, MSCs from AF and placentas showed different fluorescence intensity, reflecting the heterogeneity of these tissues. The gene expression of OCT-4, SOX-2, NANOG was not significantly different among the three groups. In AF, REX-1 and PAX-6 showed a higher expression in comparison to CV. MSCs of different extra-embryonic tissues showed no differences in immunophenotype when collected from second confluence cultures. The expression of OCT-4, NANOG and SOX-2 was not significantly different, demonstrating that all fetal sources are suitable for obtaining MSCs. These results open new possibilities for the clinical use of MSCs derived from easily accessible sources, in order to develop new protocols for clinical and experimental research. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Isolation and culture of rabbit embryonic stem cells.

PubMed

Honda, Arata

2013-01-01

Mammalian stem cells are invaluable research resources for the study of cell and embryonic development as well as practical tools for use in the production of genetically engineered animals and further therapeutics. It is important that we further our knowledge and understanding of a variety of stem cells from several different animal species before trials in humans commence. Here we describe methods for establishing rabbit embryonic stem (rES) cell lines with indefinite proliferation potential. rES cells attain maximum proliferation potential when cultured at a feeder cell density of one-sixth of that of full confluency. Higher and lower densities of feeder cells induced ES cell differentiation or division arrest. Fibroblast growth factor (FGF)2 can maintain the undifferentiated status of rES cells; however leukemia inhibitory factor (LIF) is dispensable. Under optimized conditions, rES cells could be passaged by trypsinization 50 times. This culture system enabled efficient gene transduction and clonal expansion from single cells. rES cells grew as flat monolayer cell colonies, as reported for monkey and human ES cells, and expressed pluripotency markers. Embryoid bodies and teratomas formed readily in vitro and in vivo, respectively. Characterization of ES cells from different species is important for establishing common features of pluripotency. We have demonstrated the similarity of ES cells between rabbit and humans. These cell lines could be applied directly using gene-targeting techniques, or in combination with induced pluripotent stem cells. Thus, rES cells are a suitable model for studying human transplantation therapy and disease treatments.

The moving confluence route technology with WAD scheme for 3D hydrodynamic simulation in high altitude inland waters

NASA Astrophysics Data System (ADS)

Wang, Yonggui; Yang, Yinqun; Chen, Xiaolong; Engel, Bernard A.; Zhang, Wanshun

2018-04-01

For three-dimensional hydrodynamic simulations in inland waters, the rapid changes with moving boundary and various input conditions should be considered. Some models are developed with moving boundary but the dynamic change of discharges is unresolved or ignored. For better hydrodynamic simulation in inland waters, the widely used 3D model, ECOMSED, has been improved by moving confluence route (MCR) method with a wetting and drying scheme (WAD). The fixed locations of water and pollutants inputs from tributaries, point sources and non-point sources have been changed to dynamic confluence routes as the boundary moving. The improved model was applied in an inland water area, Qingshuihai reservoir, Kunming City, China, for a one-year hydrodynamic simulation. The results were verified by water level, flow velocity and water mass conservation. Detailed water level variation analysis and velocity field comparison at different times showed that the improved model has better performance for simulating the boundary moving phenomenon and moving discharges along with water level changing than the original one. The improved three-dimensional model is available for hydrodynamics simulation in water bodies where water boundary shifts along with change of water level and have various inlets.

Confluence or independence of microwave plasma bullets in atmospheric argon plasma jet plumes

NASA Astrophysics Data System (ADS)

Li, Ping; Chen, Zhaoquan; Mu, Haibao; Xu, Guimin; Yao, Congwei; Sun, Anbang; Zhou, Yuming; Zhang, Guanjun

2018-03-01

Plasma bullet is the formation and propagation of a guided ionization wave (streamer), normally generated in atmospheric pressure plasma jet (APPJ). In most cases, only an ionization front produces in a dielectric tube. The present study shows that two or three ionization fronts can be generated in a single quartz tube by using a microwave coaxial resonator. The argon APPJ plumes with a maximum length of 170 mm can be driven by continuous microwaves or microwave pulses. When the input power is higher than 90 W, two or three ionization fronts propagate independently at first; thereafter, they confluence to form a central plasma jet plume. On the other hand, the plasma bullets move independently as the lower input power is applied. For pulsed microwave discharges, the discharge images captured by a fast camera show the ionization process in detail. Another interesting finding is that the strongest lightening plasma jet plumes always appear at the shrinking phase. Both the discharge images and electromagnetic simulations suggest that the confluence or independent propagation of plasma bullets is resonantly excited by the local enhanced electric fields, in terms of wave modes of traveling surface plasmon polaritons.

White Paper on the Use of Team Calendars with the JIRA Issue Tracking System and Confluence Collaboration Tools for the xLPR Project

DOE Office of Scientific and Technical Information (OSTI.GOV)

Klasky, Hilda B.; Williams, Paul T.; Bass, Bennett Richard

ORNL was tasked by xLPR project management to propose a team calendar for use within the xLPR consortium. Among various options that were considered, the approach judged by ORNL to best fit the needs of the xLPR project is presented in this document. The Atlassian Team Calendars plug-in used with the Confluence collaboration tool was recommended for several reasons, including the advantage that it provides for a tight integration between Confluence (found at https://xlpr.ornl.gov/wiki ) and xLPR s JIRA issue tracking system (found at https://xlpr.ornl.gov/jira ). This document is divided into two parts. The first part (Sections 1-6) consists ofmore » the white paper, which highlights some of the ways that Team Calendars can improve com mun ication between xLPR project managers, group leads, and team members when JIRA is applied for both issue tracking and change-management activities. Specific points emphasized herein are as follows: The Team Calendar application greatly enhances the added value that the JIRA and Confluence tools bring to the xLPR Project. The Team Calendar can improve com mun ication between xLPR project managers, group leads, and team members when JIRA is applied for both issue tracking and change-management activities. The Team Calendar works across different email tools such as Outlook 2011, Outlook 2010, Outlook 2007, Google Calendars and Mac s iCalendar to name a few. xLPR users can now access the wiki Confluence (with embedded Team Calendars) directly from JIRA without having to re-validate their login. The second part consists of an Annex (Section 7), which describes how users can subscribe to Team Calendars from different calendar applications. Specific instructions are given in the Annex that describe how to Import xLPR Team Calendar to Outlook Version Office 2010 Import xLPR Team Calendar to Outlook Version Office 2007 Subscribe to Team Calendar from Google Calendar The reader is directed to Section 4 for instructions on adding events to the Team Calendar or accessing ORNL staff for assistance with such additions. To seek help with your questions and problems regarding the content of this document, please contact Hilda Klasky at klaskyhb@ornl.gov« less

Suspended Sediment Loads and Tributary Inputs in the Mississippi River below St. Louis, MO, 1990-2013 Compared With Earlier Results

NASA Astrophysics Data System (ADS)

Allison, M. A.; Biedenharn, D. S.; Dahl, T. A.; Kleiss, B.; Little, C. D.

2017-12-01

Annual suspended sediment loads and water discharges were calculated in the Mississippi River mainstem channel, and at the most downstream gaging station for major tributaries, from below the Missouri confluence near St. Louis, MO to Belle Chasse, LA, as well as down the Atchafalaya distributary for water years 1990 to 2013. The purpose of the present study was to assess changes in the Mississippi River sediment budget over the past half century, and to examine the continuing role that anthropogenic (e.g., dams, river control works, soil conservation practices) and natural (e.g., rainfall and denudation rates) factors have in controlling these changes. Sixteen of the 17 measured Mississippi River tributaries decreased in total suspended sediment load) from 1970-1978 to 1990-2013. The largest decreases occurred in the 2nd (Ohio River, 41% of 1970-1978) and 4th (Arkansas River, 45% of 1970-1978) largest water sources to the Mississippi. The Missouri River remains the largest Mississippi River tributary in terms of average annual suspended sediment flux; its relative contribution increased from 38% to 51% of the total flux from the 17 measured tributaries, even as its total suspended flux declined by 13%. Averaged over the period of study (WY 1990-2013), water flux increased by 468% and sediment flux increased by 37,418% downstream from the Gavin's Point Dam to the confluence with the Mississippi. Possible reasons for this disproportional increase in suspended sediment load downstream include sediment-rich contributions from 2nd order rivers below the dams and channel incision. Suggested station improvements to the system include improved monitoring of the Upper Mississippi and Arkansas River tributaries, establishing additional mainstem stations in the reach between Thebes, IL and Arkansas City, AR, and standardization of laboratory and field methodologies to eliminate a major source of station-to-station and time-series variability in the sediment budgeting.

Upstream movements of Atlantic Salmon in the Lower Penobscot River, Maine following two dam removals and fish passage modifications

USGS Publications Warehouse

Izzo, Lisa K.; Maynard, George A.; Zydlewski, Joseph D.

2016-01-01

The Penobscot River Restoration Project (PRRP), to be completed in 2016, involved an extensive plan of dam removal, increases in hydroelectric capacity, and fish passage modifications to increase habitat access for diadromous species. As part of the PRRP, Great Works and Veazie dams were removed, making Milford Dam the first impediment to federally endangered Atlantic Salmon Salmo salar. Upstream habitat access for Atlantic Salmon is dependent upon successful and timely passage at Milford Dam because nearly all suitable spawning habitat is located upstream. In 2014 and 2015, a total of 73 adult salmon were radio-tagged to track their upstream movements through the Penobscot River to assess potential delays at (1) the dam remnants, (2) the confluence of the Stillwater Branch and the main stem of the Penobscot River below the impassable Orono Dam, and (3) the Milford Dam fish lift (installed in 2014). Movement rates through the dam remnants and the Stillwater confluence were comparable to open river reaches. Passage efficiency of the fish lift was high in both years (95% and 100%). However, fish experienced long delays at Milford Dam, with approximately one-third of fish taking more than a week to pass in each year, well below the Federal Energy Regulatory Commission passage standard of 95% within 48 h. Telemetry indicates most fish locate the fishway entrance within 5 h of arrival and were observed at the entrance at all hours of the day. These data indicate that overall transit times through the lower river were comparable to reported movement rates prior to changes to the Penobscot River due to the substantial delays seen at Milford Dam. The results of this study show that while adult Atlantic Salmon locate the new fish lift entrance quickly, passage of these fish was significantly delayed under 2014–2015 operations.

In vitro evaluation of crosslinked electrospun fish gelatin scaffolds.

PubMed

Gomes, S R; Rodrigues, G; Martins, G G; Henriques, C M R; Silva, J C

2013-04-01

Gelatin from cold water fish skin was electrospun, crosslinked and investigated as a substrate for the adhesion and proliferation of cells. Gelatin was first dissolved in either water or concentrated acetic acid and both solutions were successfully electrospun. Cross-linking was achieved via three different routes: glutaraldehyde vapor, genipin and dehydrothermal treatment. Solution's properties (surface tension, electrical conductivity and viscosity) and scaffold's properties (chemical bonds, weight loss and fiber diameters) were measured. Cellular viability was analyzed culturing 3T3 fibroblasts plated on the scaffolds and grown up to 7 days. The cells were fixed and observed with SEM or stained for DNA and F-actin and observed with confocal microscopy. In all scaffolds, the cells attached and spread with varying degrees. The evaluation of cell viability showed proliferation of cells until confluence in scaffolds crosslinked by glutaraldehyde and genipin; however the rate of growth in genipin crosslinked scaffolds was slow, recovering only by day five. The results using the dehydrothermal treatment were the less satisfactory. Our results show that glutaraldehyde treated fish gelatin is the most suitable substrate, of the three studied, for fibroblast adhesion and proliferation. Copyright © 2012 Elsevier B.V. All rights reserved.

Flood-inundation maps for Suwanee Creek from the confluence of Ivy Creek to the Noblin Ridge Drive bridge, Gwinnett County, Georgia

USGS Publications Warehouse

Musser, Jonathan W.

2012-01-01

Digital flood-inundation maps for a 6.9-mile reach of Suwanee Creek, from the confluence of Ivy Creek to the Noblin Ridge Drive bridge, were developed by the U.S. Geological Survey (USGS) in cooperation with Gwinnett County, Georgia. The inundation maps, which can be accessed through the USGS Flood Inundation Mapping Science Web site at http://water.usgs.gov/osw/flood_inundation/, depict estimates of the areal extent and depth of flooding corresponding to selected water levels (stages) at the USGS streamgage at Suwanee Creek at Suwanee, Georgia (02334885). Current stage at this USGS streamgage may be obtained at http://waterdata.usgs.gov/ and can be used in conjunction with these maps to estimate near real-time areas of inundation. The National Weather Service (NWS) is incorporating results from this study into the Advanced Hydrologic Prediction Service (AHPS) flood-warning system (http://water.weather.gov/ahps/). The NWS forecasts flood hydrographs at many places that commonly are collocated at USGS streamgages. The forecasted peak-stage information for the USGS streamgage at Suwanee Creek at Suwanee (02334885), available through the AHPS Web site, may be used in conjunction with the maps developed in this study to show predicted areas of flood inundation. A one-dimensional step-backwater model was developed using the U.S. Army Corps of Engineers HEC-RAS software for Suwanee Creek and was used to compute flood profiles for a 6.9-mile reach of the creek. The model was calibrated using the most current stage-discharge relations at the Suwanee Creek at Suwanee streamgage (02334885). The hydraulic model was then used to determine 19 water-surface profiles for flood stages at the Suwanee Creek streamgage at 0.5-foot intervals referenced to the streamgage. The profiles ranged from just above bankfull stage (7.0 feet) to approximately 1.7 feet above the highest recorded water level at the streamgage (16.0 feet). The simulated water-surface profiles were then combined with a geographic information system digital elevation model - derived from light detection and ranging (LiDAR) data having a 5.0-foot horizontal resolution - to delineate the area flooded for each 0.5-foot increment of stream stage. The availability of these maps, when combined with real-time stage information from USGS streamgages and forecasted stream stage from the NWS, provides emergency management personnel and residents with critical information during flood-response activities, such as evacuations and road closures, as well as for post-flood recovery efforts.

Occurrence and sources of Escherichia coli in metropolitan St. Louis streams, October 2004 through September 2007

USGS Publications Warehouse

Wilkison, Donald H.; Davis, Jerri V.

2010-01-01

The occurrence and sources of Escherichia coli (E. coli), one of several fecal indicator bacteria, in metropolitan St. Louis streams known to receive nonpoint source runoff, occasional discharges from combined and sanitary sewers, and treated wastewater effluent were investigated from October 2004 through September 2007. Three Missouri River sites, five Mississippi River sites, and six small basin tributary stream sites were sampled during base flow and storm events for the presence of E. coli and their sources. E. coli host-source determinations were conducted using local library based genotypic methods. Human fecal contamination in stream samples was additionally confirmed by the presence of Bacteroides thetaiotaomicron, an anaerobic, enteric bacterium with a high occurrence in, and specificity to, humans. Missouri River E. coli densities and loads during base flow were approximately 10 times greater than those in the Mississippi River above its confluence with the Missouri River. Although substantial amounts of E. coli originated from within the study area during base flow and storm events, considerable amounts of E. coli in the Missouri River, as well as in the middle Mississippi River sections downstream from its confluence with the Missouri River, originated in Missouri River reaches upstream from the study area. In lower Mississippi River reaches, bacteria contributions from the numerous combined and sanitary sewer overflows within the study area, as well as contributions from nonpoint source runoff, greatly increased instream E. coli densities. Although other urban factors cannot be discounted, average E. coli densities in streams were strongly correlated with the number of upstream combined and sanitary sewer overflow points, and the percentage of upstream impervious cover. Small basin sites with the greatest number of combined and sanitary sewer overflows (Maline Creek and the River des Peres) had larger E. coli densities, larger loads, and a greater percentage of E. coli attributable to humans than other small basin sites; however, even though small basin E. coli densities typically were much larger than in large river receiving streams, small basins contributed, on average, only a small part (a maximum of 16 percent) of the total E. coli load to larger rivers. On average, approximately one-third of E. coli in metropolitan St. Louis streams was identified as originating from humans. Another one-third of the E. coli was determined to have originated from unidentified sources; dogs and geese contributed lesser amounts, 10 and 20 percent, of the total instream bacteria. Sources of E. coli were largely independent of hydrologic conditions-an indication that sources remained relatively consistent with time.

A Novel Method for Assessing Sex-Specific and Genotype-Specific Response to Injury in Astrocyte Culture

PubMed Central

Liu, Mingyue; Oyarzabal, Esteban; Yang, Rui; Murphy, Stephanie J; Hurn, Patricia D.

2008-01-01

Female astrocytes sustain less cell death from oxygen-glucose deprivation (OGD) than male astrocytes. Arimidex, an aromatase inhibitor, abolishes these sex differences. To verify sex-dependent differences in P450 aromatase function in astrocyte cell death following OGD, we developed a novel method that uses sex-specific and genotype-specific single pup primary astrocyte cultures from wild-type (WT) and aromatase-knockout (ArKO) mice. After determining sex by external and internal examination as well as PCR and genotype by PCR amplification of tail cDNA, we established cultures from 1−3 day-old male and female, WT and ArKO mice pups and grew them to confluence in estrogen-free media. Cell death was measured by lactate dehydrogenase (LDH) assay. Our study shows that, while WT female astrocytes are more resistant to OGD than WT male cells, sex differences disappear in ArKO cells. Cell death is significantly increased in ArKO compared to WT in female astrocytes but not male cells. Therefore, P450 aromatase appears to be essential in endogenous neuroprotection in females, and this finding may have clinical implications. This innovative technique may also be applied to other in vitro studies of sex-related functional differences. PMID:18436308

Pattern Classification of Endocervical Adenocarcinoma: Reproducibility and Review of Criteria

PubMed Central

Rutgers, Joanne K.L.; Roma, Andres; Park, Kay; Zaino, Richard J.; Johnson, Abbey; Alvarado, Isabel; Daya, Dean; Rasty, Golnar; Longacre, Teri; Ronnett, Brigitte; Silva, Elvio

2017-01-01

Previously, our international team proposed a 3-tiered pattern classification (Pattern Classification) system for endocervical adenocarcinoma of the usual type that correlates with nodal disease and recurrence. Pattern Classification- A have well demarcated glands lacking destructive stromal invasion or lymphovascular invasion (lymphovascular invasion), Pattern Classification- B show localized, limited destructive invasion arising from A-type glands, and Pattern Classification- C have diffuse destructive stromal invasion, significant (filling a 4× field) confluence, or solid architecture. 24 Pattern Classification-A, 22 Pattern Classification-B, 38 Pattern Classification-C from the tumor set used in the original description were chosen using the reference diagnosis (reference diagnosis) originally established. 1 H&E slide per case was reviewed by 7 gynecologic pathologists, 4 from the original study. Kappa statistics were prepared, and cases with discrepancies reviewed. We found a majority agreement with reference diagnosis in 81% of cases, with complete or near complete (6 of 7) agreement in 50%. Overall concordance was 74%. Overall Kappa (agreement among pathologists) was .488 (moderate agreement). Pattern Classification- B has lowest kappa, and agreement is not improved by combining B+C. 6 of 7 reviewers had substantial agreement by weighted kappas (>.6), with one reviewer accounting for the majority of cases under or overcalled by 2 tiers. Confluence filling a 4× field, labyrinthine glands, or solid architecture accounted for undercalling other reference diagnosis-C cases. Missing a few individually infiltrative cells was the most common cause of undercalling reference diagnosis- B. Small foci of inflamed, loose or desmoplastic stroma lacking infiltrative tumor cells in reference diagnosis-A appeared to account for those cases up-graded to Pattern Classification-B. In summary, an overall concordance of 74% indicates that the criteria can be reproducibly applied by gynecologic pathologists. Further refinement of criteria should allow use of this powerful classification system to delineate which cervical adenocarcinomas can be safely treated conservatively. PMID:27255163

Binding of human endothelium to Ulex europaeus I-coated Dynabeads: application to the isolation of microvascular endothelium.

PubMed

Jackson, C J; Garbett, P K; Nissen, B; Schrieber, L

1990-06-01

A major problem encountered when isolating human microvascular endothelium is the presence of contaminating cells such as fibroblasts that rapidly over-grow the endothelial cells. We describe here a simple, rapid technique for purifying endothelial cells derived from the microvasculature of neonatal foreskin and osteoarthritic and rheumatoid arthritic synovium. This technique is based on the selective binding of the lectin Ulex europaeus I (UEA I) to the endothelial cell surface via fucose residues. Initially UEA I was covalently bound to tosyl-activated super-paramagnetic polystyrene beads (Dynabeads) by incubation for 24 h at room temperature. Cells were isolated by extracting microvascular segments from enzyme-treated (trypsin and Pronase) cubes of tissue. The mixed population of cells obtained were purified by incubating them at 4 degrees C for 10 min with the UEA I-coated Dynabeads. Endothelium bound to the beads whilst contaminating cells were removed by five washes with HBSS using a magnetic particle concentrator. The endothelial cells thus obtained grew to confluence as a cobblestone-like monolayer and expressed von Willebrand factor antigen. The cells were released from the Dynabeads by the competitive binding of fucose (10 min at 4 degrees C). This new method is simple and reproducible and allows pure human microvascular endothelial cells to be cultured within 2 h of obtaining a specimen.

Efficient gene transfer into nondividing cells by adeno-associated virus-based vectors.

PubMed Central

Podsakoff, G; Wong, K K; Chatterjee, S

1994-01-01

Gene transfer vectors based on adeno-associated virus (AAV) are emerging as highly promising for use in human gene therapy by virtue of their characteristics of wide host range, high transduction efficiencies, and lack of cytopathogenicity. To better define the biology of AAV-mediated gene transfer, we tested the ability of an AAV vector to efficiently introduce transgenes into nonproliferating cell populations. Cells were induced into a nonproliferative state by treatment with the DNA synthesis inhibitors fluorodeoxyuridine and aphidicolin or by contact inhibition induced by confluence and serum starvation. Cells in logarithmic growth or DNA synthesis arrest were transduced with vCWR:beta gal, an AAV-based vector encoding beta-galactosidase under Rous sarcoma virus long terminal repeat promoter control. Under each condition tested, vCWR:beta Gal expression in nondividing cells was at least equivalent to that in actively proliferating cells, suggesting that mechanisms for virus attachment, nuclear transport, virion uncoating, and perhaps some limited second-strand synthesis of AAV vectors were present in nondividing cells. Southern hybridization analysis of vector sequences from cells transduced while in DNA synthetic arrest and expanded after release of the block confirmed ultimate integration of the vector genome into cellular chromosomal DNA. These findings may provide the basis for the use of AAV-based vectors for gene transfer into quiescent cell populations such as totipotent hematopoietic stem cells. Images PMID:8057446

Efficient gene transfer into nondividing cells by adeno-associated virus-based vectors.

PubMed

Podsakoff, G; Wong, K K; Chatterjee, S

1994-09-01

Gene transfer vectors based on adeno-associated virus (AAV) are emerging as highly promising for use in human gene therapy by virtue of their characteristics of wide host range, high transduction efficiencies, and lack of cytopathogenicity. To better define the biology of AAV-mediated gene transfer, we tested the ability of an AAV vector to efficiently introduce transgenes into nonproliferating cell populations. Cells were induced into a nonproliferative state by treatment with the DNA synthesis inhibitors fluorodeoxyuridine and aphidicolin or by contact inhibition induced by confluence and serum starvation. Cells in logarithmic growth or DNA synthesis arrest were transduced with vCWR:beta gal, an AAV-based vector encoding beta-galactosidase under Rous sarcoma virus long terminal repeat promoter control. Under each condition tested, vCWR:beta Gal expression in nondividing cells was at least equivalent to that in actively proliferating cells, suggesting that mechanisms for virus attachment, nuclear transport, virion uncoating, and perhaps some limited second-strand synthesis of AAV vectors were present in nondividing cells. Southern hybridization analysis of vector sequences from cells transduced while in DNA synthetic arrest and expanded after release of the block confirmed ultimate integration of the vector genome into cellular chromosomal DNA. These findings may provide the basis for the use of AAV-based vectors for gene transfer into quiescent cell populations such as totipotent hematopoietic stem cells.

The relationship between in vitro cellular aging and in vivo human age.

PubMed Central

Schneider, E L; Mitsui, Y

1976-01-01

Differences between early and late passage cell cultures on the organelle and macromolecular levels have been attributed to cellular "aging". However, concern has been expressed over whether changes in diploid cell populations after serial passage in vitro accurately reflect human cellular aging in vivo. Studies were therefore undertaken to determine if significant differences would be observed in the in vitro lifespans of skin fibroblast cultures from old and young normal, non-hospitalized volunteers and to examine if parameters that change with in vitro "aging" are altered as a function of age in vivo. Statistically signigificant (P less than 0.05) decreases were found in the rate of fibroblast migration, onset of cell culture senescence, in vitro lifespan, cell population replication rate, and cell number at confluency of fibroblast cultures derived from the old donor group when compared to parallel cultures from young donors. No significant differences were observed in modal cell volumes and cellular macromolecular contents. The differences observed in cell cultures from old and young donors were quantitatively and qualitatively distinct from those cellular alterations observed in early and late passage WI-38 cells (in vitro "aging"). Therefore, although early and late passage cultures of human diploid cells may provide an important cell system for examining loss of replicative potential, fibroblast cultures derived from old and young human donors may be a more appropriate model system for studying human cellular aging. PMID:1068470

Assessment of dissolved-solids loading to the Colorado River in the Paradox Basin between the Dolores River and Gypsum Canyon, Utah

USGS Publications Warehouse

Shope, Christopher L.; Gerner, Steven J.

2014-01-01

Salinity loads throughout the Colorado River Basin have been a concern over recent decades due to adverse impacts on population, natural resources, and regional economics. With substantial financial resources and various reclamation projects, the salt loading to Lake Powell and associated total dissolved-solids concentrations in the Lower Colorado River Basin have been substantially reduced. The Colorado River between its confluence with the Dolores River and Lake Powell traverses a physiographic area where saline sedimentary formations and evaporite deposits are prevalent. However, the dissolved-solids loading in this area is poorly understood due to the paucity of water-quality data. From 2003 to 2011, the U.S. Geological Survey in cooperation with the U.S. Bureau of Reclamation conducted four synoptic sampling events to quantify the salinity loading throughout the study reach and evaluate the occurrence and impacts of both natural and anthropogenic sources. The results from this study indicate that under late-summer base-flow conditions, dissolved-solids loading in the reach is negligible with the exception of the Green River, and that variations in calculated loads between synoptic sampling events are within measurement and analytical uncertainties. The Green River contributed approximately 22 percent of the Colorado River dissolved-solids load, based on samples collected at the lower end of the study reach. These conclusions are supported by water-quality analyses for chloride and bromide, and the results of analyses for the stable isotopes of oxygen and deuterium. Overall, no significant sources of dissolved-solids loading from tributaries or directly by groundwater discharge, with the exception of the Green River, were identified in the study area.

Relation between Streamflow of Swiftcurrent Creek, Montana, and the Geometry of Passage for Bull Trout (Salvelinus confluentus)

USGS Publications Warehouse

Auble, Gregor T.; Holmquist-Johnson, Christopher L.; Mogen, Jim T.; Kaeding, Lynn R.; Bowen, Zachary H.

2009-01-01

Operation of Sherburne Dam in northcentral Montana has typically reduced winter streamflow in Swiftcurrent Creek downstream of the dam and resulted in passage limitations for bull trout (Salvelinus confluentus). We defined an empirical relation between discharge in Swiftcurrent Creek between Sherburne Dam and the downstream confluence with Boulder Creek and fish passage geometry by considering how the cross-sectional area of water changed as a function of discharge at a set of cross sections likely to limit fish passage. With a minimum passage window of 15 x 45 cm, passage at the cross sections increased strongly with discharge over the range of 1.2 to 24 cfs. Most cross sections did not satisfy the minimum criteria at 1.2 cfs, 25 percent had no passage at 12.7 cfs, whereas at 24 cfs all but one of 26 cross sections had some passage and 90 percent had more than 3 m of width satisfying the minimum criteria. Sensitivity analysis suggests that the overall results are not highly dependent on exact dimensions of the minimum passage window. Combining these results with estimates of natural streamflow in the study reach further suggests that natural streamflow provided adequate passage at some times in most months and locations in the study reach, although not for all individual days and locations. Limitations of our analysis include assumptions about minimum passage geometry, measurement error, limitations of the cross-sectional model we used to characterize passage, the relation of Sherburne Dam releases to streamflow in the downstream study reach in the presence of ephemeral accretions, and the relation of passage geometry as we have measured it to fish responses of movement, stranding, and mortality, especially in the presence of ice cover.

Spontaneous Packaging and Hypothermic Storage of Mammalian Cells with a Cell-Membrane-Mimetic Polymer Hydrogel in a Microchip.

PubMed

Xu, Yan; Mawatari, Kazuma; Konno, Tomohiro; Kitamori, Takehiko; Ishihara, Kazuhiko

2015-10-21

Currently, continuous culture/passage and cryopreservation are two major, well-established methods to provide cultivated mammalian cells for experiments in laboratories. Due to the lack of flexibility, however, both laboratory-oriented methods are unable to meet the need for rapidly growing cell-based applications, which require cell supply in a variety of occasions outside of laboratories. Herein, we report spontaneous packaging and hypothermic storage of mammalian cells under refrigerated (4 °C) and ambient conditions (25 °C) using a cell-membrane-mimetic methacryloyloxyethyl phosphorylcholine (MPC) polymer hydrogel incorporated within a glass microchip. Its capability for hypothermic storage of cells was comparatively evaluated over 16 days. The results reveal that the cytocompatible MPC polymer hydrogel, in combination with the microchip structure, enabled hypothermic storage of cells with quite high viability, high intracellular esterase activity, maintained cell membrane integrity, and small morphological change for more than 1 week at 4 °C and at least 4 days at 25 °C. Furthermore, the stored cells could be released from the hydrogel and exhibited the ability to adhere to a surface and achieve confluence under standard cell culture conditions. Both hypothermic storage conditions are ordinary flexible conditions which can be easily established in places outside of laboratories. Therefore, cell packaging and storage using the hydrogel incorporated within the microchip would be a promising miniature and portable solution for flexible supply and delivery of small amounts of cells from bench to bedside.

Migration and survival of Atlantic salmon Salmo salar smolts in a large natural lake.

PubMed

Kennedy, R J; Rosell, R; Millane, M; Doherty, D; Allen, M

2018-06-08

An investigation with acoustic telemetry of the passage of Salmo salar smolts through a large natural lake found heavy mortality occurred at the river-to-lake confluences (mean 31 . 2 % km -1 ), but was lower in the main body of the lake (mean 2 . 4 % km -1 ). Predation was a significant pressure on emigrating smolts as tagged pike Esox lucius aggregated at river-to-lake confluences during the peak of the smolt run. Tagged smolts mainly emmigrated into the lake in the late evening after dusk, possibly as a predator-avoidance behaviour. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Potential Role of S100A8 in Cutaneous Squamous Cell Carcinoma Differentiation.

PubMed

Shin, Jung-Min; Chang, In-Kyu; Lee, Young-Ho; Yeo, Min-Kyung; Kim, Jin-Man; Sohn, Kyung-Cheol; Im, Myung; Seo, Young-Joon; Kim, Chang-Deok; Lee, Jeung-Hoon; Lee, Young

2016-04-01

S100A8 is differentially expressed in various cell types and is associated with a number of malignant disorders. S100A8 may affect tumor biology. However, its role in cutaneous squamous cell carcinoma (SCC) is not well established. This study aims to investigate the relationship between S100A8 and cutaneous SCC development. We performed immunohistochemical staining to detect S100A8 expression in facial skin specimens of premalignant actinic keratosis (AK), malignant SCC, and normal tissues. In addition, we utilized postconfluence and high calcium-induced differentiation in a culture system model. Furthermore, we constructed a recombinant adenovirus expressing GFP-tagged S100A8 to investigate the role of S100A8 in SCC cell differentiation. S100A8 was significantly overexpressed in human cutaneous SCC compared to that in normal and AK tissues. S100A8 was gradually upregulated in SCC cells in a post-confluence-induced differentiation model. Overexpression of S100A8 in SCC cells induced by adenoviral transduction led to increased expression levels of differentiation markers, such as loricrin, involucrin, and filaggrin. S100A8 overexpression also increased loricrin and involucrin luciferase activity. S100A8 regulates cutaneous SCC differentiation and induces well-differentiated SCC formation in skin.

Skin cell isolation and expansion for cell transplantation is limited in patients using tobacco, alcohol, or are exhibiting diabetes mellitus.

PubMed

Johnen, Christa; Hartmann, Bernd; Steffen, Ingo; Bräutigam, Kirsten; Witascheck, Tom; Toman, Nidal; Küntscher, Markus V; Gerlach, Jörg C

2006-03-01

The aim of this exploratory study was to investigate the isolation and expansion of keratinocytes and fibroblasts from donors with certain medical histories. Biopsies were taken from donors (N=32) falling into one or more of the following categories: a history of heavy smoking and/or alcohol abuse, drug abuse, diabetes mellitus or steroid treatment. Cells from donors who did not fall into any of the above-mentioned categories were used as controls. Proliferation and growth behaviour of cells were analyzed by measurement of passage duration, absorbance (MTT-assay) and light microscopy. Donors with a specific medical history required larger biopsy areas than the control group for isolating a sufficient number of fibroblasts and keratinocytes. Times to confluence were significantly prolonged and absorbances (MTT) were significantly reduced in several donor groups when compared to control cultures. Biopsies from donors with steroid treatment, drug abuse and combined nicotine and alcohol abuse could not be established beyond passage 0 degrees or 1 degree, respectively. We conclude that isolation and expansion of skin cells from donors with certain medical histories may require larger biopsies, prolonged expansion times or may even result in failure. These findings may therefore be of clinical importance in the field of autologous skin cell transplantation.

BIOFILM FORMATION OF Vibrio cholerae ON STAINLESS STEEL USED IN FOOD PROCESSING.

PubMed

Fernández-Delgado, Milagro; Rojas, Héctor; Duque, Zoilabet; Suárez, Paula; Contreras, Monica; García-Amado, M Alexandra; Alciaturi, Carlos

2016-01-01

Vibrio cholerae represents a significant threat to human health in developing countries. This pathogen forms biofilms which favors its attachment to surfaces and its survival and transmission by water or food. This work evaluated the in vitro biofilm formation of V. cholerae isolated from clinical and environmental sources on stainless steel of the type used in food processing by using the environmental scanning electron microscopy (ESEM). Results showed no cell adhesion at 4 h and scarce surface colonization at 24 h. Biofilms from the environmental strain were observed at 48 h with high cellular aggregations embedded in Vibrio exopolysaccharide (VPS), while less confluence and VPS production with microcolonies of elongated cells were observed in biofilms produced by the clinical strain. At 96 h the biofilms of the environmental strain were released from the surface leaving coccoid cells and residual structures, whereas biofilms of the clinical strain formed highly organized structures such as channels, mushroom-like and pillars. This is the first study that has shown the in vitro ability of V. cholerae to colonize and form biofilms on stainless steel used in food processing.

The Nucleolus

PubMed Central

Pederson, Thoru

2011-01-01

When cells are observed by phase contrast microscopy, nucleoli are among the most conspicuous structures. The nucleolus was formally described between 1835 and 1839, but it was another century before it was discovered to be associated with a specific chromosomal locus, thus defining it as a cytogenetic entity. Nucleoli were first isolated in the 1950s, from starfish oocytes. Then, in the early 1960s, a boomlet of studies led to one of the epochal discoveries in the modern era of genetics and cell biology: that the nucleolus is the site of ribosomal RNA synthesis and nascent ribosome assembly. This epistemologically repositioned the nucleolus as not merely an aspect of nuclear anatomy but rather as a cytological manifestation of gene action—a major heuristic advance. Indeed, the finding that the nucleolus is the seat of ribosome production constitutes one of the most vivid confluences of form and function in the history of cell biology. This account presents the nucleolus in both historical and contemporary perspectives. The modern era has brought the unanticipated discovery that the nucleolus is plurifunctional, constituting a paradigm shift. PMID:21106648

The nucleolus.

PubMed

Pederson, Thoru

2011-03-01

When cells are observed by phase contrast microscopy, nucleoli are among the most conspicuous structures. The nucleolus was formally described between 1835 and 1839, but it was another century before it was discovered to be associated with a specific chromosomal locus, thus defining it as a cytogenetic entity. Nucleoli were first isolated in the 1950s, from starfish oocytes. Then, in the early 1960s, a boomlet of studies led to one of the epochal discoveries in the modern era of genetics and cell biology: that the nucleolus is the site of ribosomal RNA synthesis and nascent ribosome assembly. This epistemologically repositioned the nucleolus as not merely an aspect of nuclear anatomy but rather as a cytological manifestation of gene action-a major heuristic advance. Indeed, the finding that the nucleolus is the seat of ribosome production constitutes one of the most vivid confluences of form and function in the history of cell biology. This account presents the nucleolus in both historical and contemporary perspectives. The modern era has brought the unanticipated discovery that the nucleolus is plurifunctional, constituting a paradigm shift.

Streamflow and selenium loads during synoptic sampling of the Gunnison River and its tributaries near Delta, Colorado, November 2015

USGS Publications Warehouse

Stevens, Michael R.; Leib, Kenneth J.; Thomas, Judith C.; Bauch, Nancy J.; Richards, Rodney J.

2018-06-13

In response to the need for more information about selenium (Se) sources and transport, the U.S. Geological Survey, in cooperation with the Colorado Water Conservation Board, completed a study that characterized Se loads in a reach of the Gunnison River between Delta and Grand Junction, Colo. This report identifies where possible dissolved Se loading is occurring in a study reach in the Lower Gunnison River Basin between Delta and Grand Junction on November 19, 2015.The combined Se loads from the Gunnison River at Delta (site 3) and the Uncompahgre River at Delta (site 4) were about 95 percent of the load at the furthest downstream main-stem sample location at the Gunnison River below Roubideau Creek near Delta (site 20) (31.6 and 33.4 pounds per day, respectively), indicating that about 5 percent of the total load (1.8 pounds) was potentially contributed from diffuse groundwater inflow. Main-stem streamflow accounting during November 2015 in a downstream direction was not supportive of substantial net gains or losses in the main-stem water balance.The cumulative load from measured tributary inflows downstream from the Uncompahgre River confluence only amounted to 1.2 pounds of the main-stem loads (1.8 pounds gain) from site 4 to the end of the synoptic reach at site 20. The remaining 33 percent (about 0.6 pounds) of Se load increase was not accounted for by known tributary inflow. Yet, the small changes in the streamflow mass balance in the same reach does not strongly support a net inflow explanation for the apparent gain in load.Based on the results of the loading and streamflow analysis, when errors in the loading estimates are considered, there is no conclusive evidence of an appreciable amount of Se load that is unaccounted for in the study reach of the Gunnison River as was originally hypothesized. Differences determined from comparisons of cumulative tributary loads and Gunnison River main-stem loads for this study are within error estimates of the main-stem loads.

Water and sediment study of the Snake River watershed, Colorado, Oct. 9-12, 2001

USGS Publications Warehouse

Fey, D.L.; Church, S.E.; Unruh, D.M.; Bove, D.J.

2002-01-01

The Snake River watershed, located upstream from Dillon Reservoir in the central mountains of Colorado, has been affected by historical base-metal mining. Trout stocked in the Snake River for recreational purposes do not survive through the winter. Sediment cores analyzed by previous investigators from the reservoir revealed elevated concentrations of base metals and mercury. We collected 36 surface water samples (filtered and unfiltered) and 38 streambed-sediment samples from streams in the Snake River watershed. Analyses of the sediment and water samples show that concentrations of several metals exceed aquatic life standards in one or both media. Ribbon maps showing dissolved concentrations of zinc, cadmium, copper, and manganese in water (0.45-micron filtered and corrected for the ameliorating effect of hardness), and copper, cadmium, and zinc in sediment indicate reaches where toxic effects on trout would be expected and stream reaches where toxicity standards for rainbow, brown, and brook trout are exceeded. Instantaneous loads for sulfate, strontium, iron, cadmium, copper, and zinc were calculated from 0.45-micron-filtered water concentrations and discharge measurements were made at each site. Sulfate and strontium behave conservatively, whereas copper, cadmium, and zinc are reactive. The dissolved copper load entering the reservoir is less than 20 percent of the value calculated from some upper reaches; copper is transferred to suspended and or streambed sediment by sorption to iron oxyhydroxides. Higher percentages of zinc and cadmium reach the reservoir in dissolved form; however, load calculations indicate that some of these metals are also precipitated out of solution. The most effective remediation activities should be concentrated on reducing the dissolved loads of zinc, cadmium, and copper in two reaches of lower Peru Creek between the confluence with the Snake River and Cinnamon Gulch. We analyzed all streambed sediment for mercury and selected streambed-sediment and reservoir core samples for lead isotope signatures. Results indicate that the mercury anomaly in the reservoir sediment was not from any known source in the Snake River, Blue River, or Tenmile Creek watersheds. Its source remains an enigma.

Fish population and habitat analysis in Buck Creek, Washington, prior to recolonization by anadromous salmonids after the removal of Condit Dam

USGS Publications Warehouse

Allen, M. Brady; Burkhardt, Jeanette; Munz, Carrie; Connolly, Patrick J.

2012-01-01

We assessed the physical and biotic conditions in the part of Buck Creek, Washington, potentially accessible to anadromous fishes. This creek is a major tributary to the White Salmon River upstream of Condit Dam, which was breached in October 2011. Habitat and fish populations were characterized in four stream reaches. Reach breaks were based on stream gradient, water withdrawals, and fish barriers. Buck Creek generally was confined, with a single straight channel and low sinuosity. Boulders and cobble were the dominant stream substrate, with limited gravel available for spawning. Large-cobble riffles were 83 percent of the available fish habitat. Pools, comprising 15 percent of the surface area, mostly were formed by bedrock with little instream cover and low complexity. Instream wood averaged 6—10 pieces per 100 meters, 80 percent of which was less than 50 centimeters in diameter. Water temperature in Buck Creek rarely exceeded 16 degrees Celsius and did so for only 1 day at river kilometer (rkm) 3 and 11 days at rkm 0.2 in late July and early August 2009. The maximum temperature recorded was 17.2 degrees Celsius at rkm 0.2 on August 2, 2009. Minimum summer discharge in Buck Creek was 3.3 cubic feet per second downstream of an irrigation diversion (rkm 3.1) and 7.7 cubic feet per second at its confluence with the White Salmon River. Rainbow trout (Oncorhynchus mykiss) was the dominant fish species in all reaches. The abundance of age-1 or older rainbow trout was similar between reaches. However, in 2009 and 2010, the greatest abundance of age-0 rainbow trout (8 fish per meter) was in the most downstream reach. These analyses in Buck Creek are important for understanding the factors that may limit fish abundance and productivity, and they will help identify and prioritize potential restoration actions. The data collected constitute baseline information of pre-dam removal conditions that will allow assessment of changes in fish populations now that Condit Dam has been removed and anadromous fish have an opportunity to recolonize Buck Creek.

Movements of adult chinook salmon during spawning migration in a metals-contaminated system, Coeur d'Alene River, Idaho

USGS Publications Warehouse

Goldstein, J.N.; Woodward, D.F.; Farag, A.M.

1999-01-01

Spawning migration of adult male chinook salmon Oncorhynchus tshawytscha was monitored by radio telemetry to determine their response to the presence of metals contamination in the South Fork of the Coeur d'Alene River, Idaho. The North Fork of the Coeur d'Alene River is relatively free of metals contamination and was used as a control. In all, 45 chinook salmon were transported from their natal stream, Wolf Lodge Creek, tagged with radio transmitters, and released in the Coeur d'Alene River 2 km downstream of the confluence of the South Fork and the North Fork of the Coeur d'Alene River. Fixed telemetry receivers were used to monitor the upstream movement of the tagged chinook salmon through the confluence area for 3 weeks after release. During this period, general water quality and metals concentrations were monitored in the study area. Of the 23 chinook salmon observed to move upstream from the release site and through the confluence area, the majority (16 fish, 70%) moved up the North Fork, and only 7 fish (30%) moved up the South Fork, where greater metals concentrations were observed. Our results agree with laboratory findings and suggest that natural fish populations will avoid tributaries with high metals contamination.

Borneo vortex and meso-scale convective rainfall

NASA Astrophysics Data System (ADS)

Koseki, S.; Koh, T.-Y.; Teo, C.-K.

2013-08-01

We have investigated how the Borneo vortex develops over the equatorial South China Sea under cold surge conditions in December during the Asian winter monsoon. Composite analysis using reanalysis and satellite datasets has revealed that absolute vorticity and water vapour are transported by strong cold surges from upstream of the South China Sea to around the equator. Rainfall is correspondingly enhanced over the equatorial South China Sea. A semi-idealized experiment reproduced the Borneo vortex over the equatorial South China Sea during a "perpetual" cold surge. The Borneo vortex is manifested as a meso-α cyclone with a comma-shaped rainband in the northeast sector of the cyclone. Vorticity budget analysis showed that the growth of the meso-α cyclone was achieved mainly by vortex stretching. The comma-shaped rainband consists of clusters of meso-β scale rainfall patches. The warm and wet cyclonic southeasterly flow meets with the cold and dry northeasterly surge forming a confluence front in the northeastern sector of the cyclone. Intense upward motion and heavy rainfall result both due to the low-level convergence and the favourable thermodynamic profile at the confluence front. At both meso-α and meso-β scales, the convergence is ultimately caused by the deviatoric strain in the confluence wind pattern but is much enhanced by nonlinear self-enhancement dynamics.

Multidetector CT angiography in living donor renal transplantation: accuracy and discrepancies in right venous anatomy.

PubMed

Kulkarni, S; Emre, S; Arvelakis, A; Asch, W; Bia, M; Formica, R; Israel, G

2011-01-01

Multidetector computed tomography (MDCT) angiography is a reliable technique for assessing pre-operative renal anatomy in living kidney donors. The method has largely evolved into protocols that eliminate dedicated venous phase and instead utilize a combined arterial/venous phase to delineate arterial and venous anatomy simultaneously. Despite adoption of this protocol, there has been no study to assess its accuracy. To assess whether or not MDCT angiography compares favorably to intra-operative findings, 102 donors underwent MDCT angiography without a dedicated venous phase with surgical interpretation of renal anatomy. Anatomical variants included multiple arteries (12%), multiple veins (7%), early arterial bifurcation (13%), late venous confluence (5%), circumaortic renal veins (5%), retroaortic vein (1%), and ureteral duplication (2%). The sensitivity and specificity of multiple arterial anomalies were 100% and 97%, respectively. The sensitivity and specificity of multiple venous anomalies were 92% and 98%, respectively. The most common discrepancy was noted exclusively in the interpretation of right venous anatomy as it pertained to the renal vein/vena cava confluence (3%). MDCT angiography using a combined arterial/venous contrast-enhanced phase provides suitable depiction of renal donor anatomy. Careful consideration should be given when planning a right donor nephrectomy whether the radiographic interpretation is suggestive of a late confluence. © 2010 John Wiley & Sons A/S.

High content analysis of differentiation and cell death in human adipocytes.

PubMed

Doan-Xuan, Quang Minh; Sarvari, Anitta K; Fischer-Posovszky, Pamela; Wabitsch, Martin; Balajthy, Zoltan; Fesus, Laszlo; Bacso, Zsolt

2013-10-01

Understanding adipocyte biology and its homeostasis is in the focus of current obesity research. We aimed to introduce a high-content analysis procedure for directly visualizing and quantifying adipogenesis and adipoapoptosis by laser scanning cytometry (LSC) in a large population of cell. Slide-based image cytometry and image processing algorithms were used and optimized for high-throughput analysis of differentiating cells and apoptotic processes in cell culture at high confluence. Both preadipocytes and adipocytes were simultaneously scrutinized for lipid accumulation, texture properties, nuclear condensation, and DNA fragmentation. Adipocyte commitment was found after incubation in adipogenic medium for 3 days identified by lipid droplet formation and increased light absorption, while terminal differentiation of adipocytes occurred throughout day 9-14 with characteristic nuclear shrinkage, eccentric nuclei localization, chromatin condensation, and massive lipid deposition. Preadipocytes were shown to be more prone to tumor necrosis factor alpha (TNFα)-induced apoptosis compared to mature adipocytes. Importantly, spontaneous DNA fragmentation was observed at early stage when adipocyte commitment occurs. This DNA damage was independent from either spontaneous or induced apoptosis and probably was part of the differentiation program. © 2013 International Society for Advancement of Cytometry. Copyright © 2013 International Society for Advancement of Cytometry.

Timescale Separation of Positive and Negative Signaling Creates History-Dependent Responses to IgE Receptor Stimulation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Harmon, Brooke; Chylek, Lily A.; Liu, Yanli

The high-affinity receptor for IgE expressed on the surface of mast cells and basophils interacts with antigens, via bound IgE antibody, and triggers secretion of inflammatory mediators that contribute to allergic reactions. To understand how past inputs (memory) influence future inflammatory responses in mast cells, a microfluidic device was used to precisely control exposure of cells to alternating stimulatory and non-stimulatory inputs. We determined that the response to subsequent stimulation depends on the interval of signaling quiescence. For shorter intervals of signaling quiescence, the second response is blunted relative to the first response, whereas longer intervals of quiescence induce anmore » enhanced second response. Through an iterative process of computational modeling and experimental tests, we found that these memory-like phenomena arise from a confluence of rapid, short-lived positive signals driven by the protein tyrosine kinase Syk; slow, long-lived negative signals driven by the lipid phosphatase Ship1; and slower degradation of Ship1 co-factors. This work advances our understanding of mast cell signaling and represents a generalizable approach for investigating the dynamics of signaling systems.« less

Different sucrose-isomaltase response of Caco-2 cells to glucose and maltose suggests dietary maltose sensing

PubMed Central

Cheng, Min-Wen; Chegeni, Mohammad; Kim, Kee-Hong; Zhang, Genyi; Benmoussa, Mustapha; Quezada-Calvillo, Roberto; Nichols, Buford L.; Hamaker, Bruce R.

2014-01-01

Using the small intestine enterocyte Caco-2 cell model, sucrase-isomaltase (SI, the mucosal α-glucosidase complex) expression and modification were examined relative to exposure to different mono- and disaccharide glycemic carbohydrates. Caco-2/TC7 cells were grown on porous supports to post-confluence for complete differentiation, and dietary carbohydrate molecules of glucose, sucrose (disaccharide of glucose and fructose), maltose (disaccharide of two glucoses α-1,4 linked), and isomaltose (disaccharide of two glucoses α-1,6 linked) were used to treat the cells. qRT-PCR results showed that all the carbohydrate molecules induced the expression of the SI gene, though maltose (and isomaltose) showed an incremental increase in mRNA levels over time that glucose did not. Western blot analysis of the SI protein revealed that only maltose treatment induced a higher molecular weight band (Mw ~245 kDa), also at higher expression level, suggesting post-translational processing of SI, and more importantly a sensing of maltose. Further work is warranted regarding this putative sensing response as a potential control point for starch digestion and glucose generation in the small intestine. PMID:24426192

Different sucrose-isomaltase response of Caco-2 cells to glucose and maltose suggests dietary maltose sensing.

PubMed

Cheng, Min-Wen; Chegeni, Mohammad; Kim, Kee-Hong; Zhang, Genyi; Benmoussa, Mustapha; Quezada-Calvillo, Roberto; Nichols, Buford L; Hamaker, Bruce R

2014-01-01

Using the small intestine enterocyte Caco-2 cell model, sucrase-isomaltase (SI, the mucosal α-glucosidase complex) expression and modification were examined relative to exposure to different mono- and disaccharide glycemic carbohydrates. Caco-2/TC7 cells were grown on porous supports to post-confluence for complete differentiation, and dietary carbohydrate molecules of glucose, sucrose (disaccharide of glucose and fructose), maltose (disaccharide of two glucoses α-1,4 linked), and isomaltose (disaccharide of two glucoses α-1,6 linked) were used to treat the cells. qRT-PCR results showed that all the carbohydrate molecules induced the expression of the SI gene, though maltose (and isomaltose) showed an incremental increase in mRNA levels over time that glucose did not. Western blot analysis of the SI protein revealed that only maltose treatment induced a higher molecular weight band (Mw ~245 kDa), also at higher expression level, suggesting post-translational processing of SI, and more importantly a sensing of maltose. Further work is warranted regarding this putative sensing response as a potential control point for starch digestion and glucose generation in the small intestine.

The Weddell-Scotia Confluence in midwinter

NASA Astrophysics Data System (ADS)

Muench, Robin D.; Gunn, John T.; Husby, David M.

1990-10-01

The southern central Scotia Sea, site of the Weddell-Scotia Confluence where outflowing Weddell Sea waters converge with the eastward flowing waters of the Scotia Sea, was sampled during June-August (austral winter) 1988 with respect to temperature and salinity. Both drogued and ice-mounted drifters, tracked by Argos, were deployed in the region and yielded Lagrangian drift tracks of ice and water motion. The data substantiate past accounts of the region, based upon summer field research, as dominated by eastward flow upon which a complex array of mesoscale features is superimposed. Weddell-Scotia Confluence Water, documented by past summer work in the region and characterized by decreased static stability, was not detected, and the Scotia Front was not well defined. The region was one of intense mixing activity and primarily anticyclonic mesoscale features. Two such features, one an eddy and the other either an eddy or a meander in the Scotia Front, dominated the mesoscale field. With warm cores and containing Polar Front Water, they may have been advected eastward from Drake Passage or may have formed as detached eddies from a sharp northward bend in the Polar Front which typically lies just west of the study region. Several smaller eddies, primarily anticyclonic and some having warm cores, were also detected. There was no evidence of the deep convective mixing which has been hypothesized, on the basis of past summer data, to occur in winter, and vigorous vertical mixing was limited to a 100-m-thick upper mixed layer. Vertical stability in the upper layers was enhanced by low-salinity water derived from melting ice. Temperature-salinity analyses show that winter water in the study region can be derived through isopycnal mixing between waters from the Scotia Sea and waters from the northwestern Weddell Sea. This is in apparent contrast with summer conditions, wherein conditioning of water either through vertical mixing or via lateral mixing on continental margins has been invoked to arrive at the water mass characteristics which typify the Weddell-Scotia Confluence.

Neural correlates of target selection for reaching movements in superior colliculus

PubMed Central

McPeek, Robert M.

2014-01-01

We recently demonstrated that inactivation of the primate superior colliculus (SC) causes a deficit in target selection for arm-reaching movements when the reach target is located in the inactivated field (Song JH, Rafal RD, McPeek RM. Proc Natl Acad Sci USA 108: E1433–E1440, 2011). This is consistent with the notion that the SC is part of a general-purpose target selection network beyond eye movements. To understand better the role of SC activity in reach target selection, we examined how individual SC neurons in the intermediate layers discriminate a reach target from distractors. Monkeys reached to touch a color oddball target among distractors while maintaining fixation. We found that many SC neurons robustly discriminate the goal of the reaching movement before the onset of the reach even though no saccade is made. To identify these cells in the context of conventional SC cell classification schemes, we also recorded visual, delay-period, and saccade-related responses in a delayed saccade task. On average, SC cells that discriminated the reach target from distractors showed significantly higher visual and delay-period activity than nondiscriminating cells, but there was no significant difference in saccade-related activity. Whereas a majority of SC neurons that discriminated the reach target showed significant delay-period activity, all nondiscriminating cells lacked such activity. We also found that some cells without delay-period activity did discriminate the reach target from distractors. We conclude that the majority of intermediate-layer SC cells discriminate a reach target from distractors, consistent with the idea that the SC contains a priority map used for effector-independent target selection. PMID:25505107

Post-mitotic human dermal fibroblasts efficiently support the growth of human follicular keratinocytes.

PubMed

Limat, A; Hunziker, T; Boillat, C; Bayreuther, K; Noser, F

1989-05-01

For growth at low seeding densities, keratinocytes isolated from human tissues like epidermis or hair follicles are dependent on mesenchyme-derived feeder cells such as the 3T3-cell employed so far. As an alternative method, the present study describes the use of post-mitotic human dermal fibroblasts sublethally irradiated or mitomycin C-treated. Special emphasis was put on efficient growth of primary keratinocyte cultures plated at very low seeding densities. Thus, outer root sheath cells isolated from two anagen human hair follicles and plated in a 35-mm culture dish (3 - 6 X 10(2) attached cells) grew to confluence within 3 weeks (6 - 8 X 10(5) cells). Similar results were obtained for interfollicular keratinocytes. A crucial point for the function of these fibroblast feeder cells is plating at appropriate densities, considering their tremendous increase in cell size at the post-mitotic state. Plating densities of 4 - 5 X 10(3/cm2 allow full spreading of the feeder cells and do not impede the settling and expansion of the keratinocytes. Major advantages of this system include easier handling and better reproducibility than using 3T3-cells. Moreover, homologous fibroblast feeders mimic more closely the physiologic situation and therefore might provide a valuable tool for studying interactions between human mesenchymal and epithelial cells. Finally, potential hazards of using transformed feeder cells from a different species in keratinocyte cultures raised for wound covering in humans could be thus avoided.

An in-vitro scaffold-free epithelial-fibroblast coculture model for the larynx

PubMed Central

Walimbe, Tanaya; Panitch, Alyssa; Sivasankar, M. Preeti

2017-01-01

Objective Physiologically relevant, well-characterized in vitro vocal fold coculture models are needed to test the effects of various challenges and therapeutics on vocal fold physiology. We characterize a healthy state coculture model, created by using bronchial/tracheal epithelial cells and immortalized vocal fold fibroblasts. We also demonstrate that this model can be induced into a fibroplastic state to overexpress stress fibers using TGFβ1. Method Cell metabolic activity of immortalized human vocal fold fibroblasts incubated in different media combinations were confirmed with MTT assay. Fibroblasts were grown to confluence and primary bronchial/tracheal epithelial cells suspended in coculture media were seeded directly over the base layer of the fibroblasts. Cells were treated with TGFβ1 to induce myofibroblast formation. Cell shape and position was confirmed by live cell tracking, fibrosis was confirmed by probing for α smooth muscle actin (α-SMA) and phenotype was confirmed by immunostaining for vimentin and E-cadherin. Results Fibroblasts retain metabolic activity in coculture epithelial media. Live cell imaging revealed a layer of epithelial cells atop fibroblasts. α-SMA expression was enhanced in TGFβ1 treated cells, confirming that both cell types maintained a healthy phenotype in coculture, and can be induced into overexpressing stress fibers. Vimentin and E-cadherin immunostaining show that cells retain phenotype in coculture. Conclusion These data lay effective groundwork for a functional coculture model that retains the reproducibility necessary to serve as a viable diagnostic and therapeutic screening platform. Level of Evidence NA PMID:27859361

Preliminary assessment of channel stability and bed-material transport along Hunter Creek, southwestern Oregon

USGS Publications Warehouse

Jones, Krista L.; Wallick, J. Rose; O'Connor, Jim E.; Keith, Mackenzie K.; Mangano, Joseph F.; Risley, John C.

2011-01-01

This preliminary assessment of (1) bed-material transport in the Hunter Creek basin, (2) historical changes in channel condition, and (3) supplementary data needed to inform permitting decisions regarding instream gravel extraction revealed the following: Along the lower 12.4 km (kilometers) of Hunter Creek from its confluence with the Little South Fork Hunter Creek to its mouth, the river has confined and unconfined segments and is predominately alluvial in its lowermost 11 km. This 12.4-km stretch of river can be divided into two geomorphically distinct study reaches based primarily on valley physiography. In the Upper Study Reach (river kilometer [RKM] 12.4-6), the active channel comprises a mixed bed of bedrock, boulders, and smaller grains. The stream is confined in the upper 1.4 km of the reach by a bedrock canyon and in the lower 2.4 km by its valley. In the Lower Study Reach (RKM 6-0), where the area of gravel bars historically was largest, the stream flows over bed material that is predominately alluvial sediments. The channel alternates between confined and unconfined segments. The primary human activities that likely have affected bed-material transport and the extent and area of gravel bars are (1) historical and ongoing aggregate extraction from gravel bars in the study area and (2) timber harvest and associated road construction throughout the basin. These anthropogenic activities likely have varying effects on sediment transport and deposition throughout the study area and over time. Although assessing the relative effects of these anthropogenic activities on sediment dynamics would be challenging, the Hunter Creek basin may serve as a case study for such an assessment because it is mostly free of other alterations to hydrologic and geomorphic processes such as flow regulation, dredging, and other navigation improvements that are common in many Oregon coastal basins. Several datasets are available that may support a more detailed physical assessment of Hunter Creek. The entire study area has been captured in aerial photographs at least once per decade since the 1940s. This temporally rich photograph dataset would support quantitative analyses of changes in channel planform as well as vegetation cover. Light Detection And Ranging (LiDAR) data collected in 2008 would facilitate hydraulic and sediment-transport modeling and characterization of bar elevations throughout most of the study area. Few studies describing channel morphology and sediment transport exist for the Hunter Creek basin. The most detailed study reported channel incision and bank instability as well as the loss of point bars and pools in the lower 3.9 km of Hunter Creek from slightly downstream of its confluence with Yorke Creek to its mouth (EA Engineering, Sci-ence, and Technology, 1998). Repeat channel cross-sections collected from 1994 to 2010 at four bridges indicate that Hunter Creek is dynamic and subject to channel shifting, aggradation, and incision. Despite this dynamism, the channel at three bridge crossings showed little net change in thalweg elevation during this period. However, the channel thalweg aggraded 0.55 m from 2004 to 2008 near the bridge at RKM 3.5. Systematic delineation of gravel bars from aerial photographs collected in 1940, 1965, 2005, and 2009 indicates a 52-percent reduction in the area of bed-material sediment throughout the study area from 1940 to 2009. Net bar loss was greatest in the Lower Study Reach from RKM 1-4 and mainly is associ-ated with the encroachment of vegetation onto upper-bar surfaces lacking apparent vegetation in 1940. Bar-surface material was approximately equal in size to bar-subsurface material at Conn Creek Bar, whereas it was distinctly coarser than the subsurface material at Menasha Bar. Armoring ratios, which indicate the coarseness of the bar surface relative to the bar subsurface, were calculated as 0.97 for Conn Creek Bar and 1.5 for Menasha Bar. These ratios tentatively show that

Overview of Climate Confluence Security Issues

NASA Astrophysics Data System (ADS)

Reisman, J. P.

2011-12-01

Presentation will focus on an overview of the security perspectives based on the confluence considerations including energy, economics and climate change. This will include perspectives from reports generated by the Quadrennial Defense Review, Joint Forces Command, the Center for Strategic International Studies, MIT, the Inter-agency Climate Change Adaptation Task Force, the Central Intelligence Agency, the Center for Naval Analysis, and other relevant reports. The presentation will highlight the connections between resource issues and climate change which can be interpreted into security concerns. General discussion of global issues, contextual review of AR4 WGII may be included and any other report updates as applicable. The purpose of this presentation is to give a rounded view of the general qualitative and quantitative perspectives regarding climate related security considerations.

Design of a peptide-based vector, PepFect6, for efficient delivery of siRNA in cell culture and systemically in vivo

PubMed Central

EL Andaloussi, Samir; Lehto, Taavi; Mäger, Imre; Rosenthal-Aizman, Katri; Oprea, Iulian I.; Simonson, Oscar E.; Sork, Helena; Ezzat, Kariem; Copolovici, Dana M.; Kurrikoff, Kaido; Viola, Joana R.; Zaghloul, Eman M.; Sillard, Rannar; Johansson, Henrik J.; Said Hassane, Fatouma; Guterstam, Peter; Suhorutšenko, Julia; Moreno, Pedro M. D.; Oskolkov, Nikita; Hälldin, Jonas; Tedebark, Ulf; Metspalu, Andres; Lebleu, Bernard; Lehtiö, Janne; Smith, C. I. Edvard; Langel, Ülo

2011-01-01

While small interfering RNAs (siRNAs) have been rapidly appreciated to silence genes, efficient and non-toxic vectors for primary cells and for systemic in vivo delivery are lacking. Several siRNA-delivery vehicles, including cell-penetrating peptides (CPPs), have been developed but their utility is often restricted by entrapment following endocytosis. Hence, developing CPPs that promote endosomal escape is a prerequisite for successful siRNA implementation. We here present a novel CPP, PepFect 6 (PF6), comprising the previously reported stearyl-TP10 peptide, having pH titratable trifluoromethylquinoline moieties covalently incorporated to facilitate endosomal release. Stable PF6/siRNA nanoparticles enter entire cell populations and rapidly promote endosomal escape, resulting in robust RNAi responses in various cell types (including primary cells), with minimal associated transcriptomic or proteomic changes. Furthermore, PF6-mediated delivery is independent of cell confluence and, in most cases, not significantly hampered by serum proteins. Finally, these nanoparticles promote strong RNAi responses in different organs following systemic delivery in mice without any associated toxicity. Strikingly, similar knockdown in liver is achieved by PF6/siRNA nanoparticles and siRNA injected by hydrodynamic infusion, a golden standard technique for liver transfection. These results imply that the peptide, in addition to having utility for RNAi screens in vitro, displays therapeutic potential. PMID:21245043

Tumor endothelial marker 5 expression in endothelial cells during capillary morphogenesis is induced by the small GTPase Rac and mediates contact inhibition of cell proliferation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vallon, Mario, E-mail: m.vallon@arcor.de; Rohde, Franziska; Janssen, Klaus-Peter

2010-02-01

Tumor endothelial marker (TEM) 5 is an adhesion G-protein-coupled receptor upregulated in endothelial cells during tumor and physiologic angiogenesis. So far, the mechanisms leading to upregulation of TEM5 and its function during angiogenesis have not been identified. Here, we report that TEM5 expression in endothelial cells is induced during capillary-like network formation on Matrigel, during capillary morphogenesis in a three-dimensional collagen I matrix, and upon confluence on a two-dimensional matrix. TEM5 expression was not induced by a variety of soluble angiogenic factors, including VEGF and bFGF, in subconfluent endothelial cells. TEM5 upregulation was blocked by toxin B from Clostridium difficile,more » an inhibitor of the small GTPases Rho, Rac, and Cdc42. The Rho inhibitor C3 transferase from Clostridium botulinum did not affect TEM5 expression, whereas the Rac inhibitor NSC23766 suppressed TEM5 upregulation. An excess of the soluble TEM5 extracellular domain or an inhibitory monoclonal TEM5 antibody blocked contact inhibition of endothelial cell proliferation resulting in multilayered islands within the endothelial monolayer and increased vessel density during capillary formation. Based on our results we conclude that TEM5 expression during capillary morphogenesis is induced by the small GTPase Rac and mediates contact inhibition of proliferation in endothelial cells.« less

Hydraulic Conductivity of Endothelial Cell-Initiated Arterial Cocultures

PubMed Central

Mathura, Rishi A.; Russell-Puleri, Sparkle; Cancel, Limary; Tarbell, John M.

2014-01-01

This study describes cocultures of arterial smooth muscle cells (SMC) and endothelial cells (EC) and the influences of their heterotypic interactions on hydraulic conductivity (Lp), an important transport property. A unique feature of these cocultures is that ECs were first grown to confluence and then SMCs were inoculated. Bovine aortic smooth muscle cells (BASMCs) and bovine aortic endothelial cells (BAECs) were cocultured on Transwell Permeable Supports, and then exposed to a pressure-driven transmural flow. Lp across each culture was measured using a bubble tracking apparatus that determined water flux (Jv). Our results indicate that arterial Lp is significantly modulated by EC-SMC proximity, and serum content in culture. The Lp of cocultures was also compared to the predictions of a resistances-in-series model to distinguish the contributions of heterotypic interactions between SMCs and ECs. Conditions that lead to significantly reduced coculture Lp, compared to BAEC monoculture controls, have been uncovered and the lowest Lp in the literature for an in-vitro system are reported. In addition, VE-cadherin immunostaining of intact BAEC monolayers in each culture configuration reveals that EC-SMC proximity on a porous membrane has a dramatic influence on EC morphology patterns. The cocultures with the lowest Lp have ECs with significantly elongated morphology. Confocal imaging indicates that there are no direct EC-SMC contacts in coculture. PMID:24264601

Hilar Inflammatory Pseudotumour with Hepatic Artery Atheroma- mimicker of Klatskin Tumour.

PubMed

Rastogi, Archana; Bihari, Chhagan; Gupta, Nalini; Deka, Pranjal; Kumar, Arvind; Negi, Sanjay Singh; Arora, Ankur

2015-03-01

Inflammatory pseudotumour of hilar biliary structures is an extremely rare benign lesion that can mimic hilar cholangiocarcinoma. Clinical presentation and imaging findings often pose diagnostic difficulties. Main histopathological findings are the presence of myofibroblastic spindle cells, plasma cells, macrophages, and lymphocytes without cellular atypia or atypical mitotic figures. We describe a case of 62 year old male who presented with surgical obstructive jaundice. Imaging revealed a mass lesion involving the biliary confluence with upstream dilatation of biliary tree. Diagnosis of hilar cholangiocarcinoma with type III hilar block was made. Intraoperately hilar mass lesion was found which was encasing right hepatic artery with no evidence of metastasis. The patient underwent Right hepatectomy with caudate lobectomy with complete common bile duct (CBD) excision with Roux en Y hepaticojejunostomy. Unexpectedly histopathological examination showed no evidence of malignancy and revealed hilar inflammatory pseudotumour with hepatic artery atherosclerosis. Preoperative imaging, operative management, pathologic diagnosis and literature review are being presented in view of rarity of the case.

BAG-1 enhances cell-cell adhesion, reduces proliferation and induces chaperone-independent suppression of hepatocyte growth factor-induced epidermal keratinocyte migration

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hinitt, C.A.M.; Wood, J.; Lee, S.S.

2010-08-01

Cell motility is important in maintaining tissue homeostasis, facilitating epithelial wound repair and in tumour formation and progression. The aim of this study was to determine whether BAG-1 isoforms regulate epidermal cell migration in in vitro models of wound healing. In the human epidermal cell line HaCaT, endogenous BAG-1 is primarily nuclear and increases with confluence. Both transient and stable p36-Bag-1 overexpression resulted in increased cellular cohesion. Stable transfection of either of the three human BAG-1 isoforms p36-Bag-1 (BAG-1S), p46-Bag-1 (BAG-1M) and p50-Bag-1 (BAG-1L) inhibited growth and wound closure in serum-containing medium. However, in response to hepatocyte growth factor (HGF)more » in serum-free medium, BAG-1S/M reduced communal motility and colony scattering, but BAG-1L did not. In the presence of HGF, p36-Bag-1 transfectants retained proliferative response to HGF with no change in ERK1/2 activation. However, the cells retained E-cadherin localisation at cell-cell junctions and exhibited pronounced cortical actin. Point mutations in the BAG domain showed that BAG-1 inhibition of motility is independent of its function as a chaperone regulator. These findings are the first to suggest that BAG-1 plays a role in regulating cell-cell adhesion and suggest an important function in epidermal cohesion.« less

Hydrogeology of Valley-Fill Aquifers and Adjacent Areas in Eastern Chemung County, New York

USGS Publications Warehouse

Heisig, Paul M.

2015-10-19

Water-resource potential is greatest within saturated sand and gravel in the Chemung River valley (nearly 1 mile wide), especially where induced infiltration of additional water from the Chemung River is possible. The second most favorable area is the Newtown Creek valley at the confluence of Newtown Creek with North Branch Newtown Creek east of Horseheads, N.Y. Extensive sand and gravel deposits within the Breesport, N.Y., area are largely unsaturated but may have greater saturation along the east side of Jackson Creek immediately north of Breesport. Till deposits confine sand and gravel along Newtown Creek at Erin, N.Y., and along much of the upper reach of North Branch Newtown Creek; this confining unit may limit recharge and potential well yield. The north-south oriented valleys of Baldwin and Wynkoop Creeks end at notched divides that imply input of glacial meltwater and limited sediment from outside of the present watersheds. These two valleys are relatively narrow but contain variably sorted sand and gravel, which, in places, may be capable of supplying modest-size community water systems.

Differences in Ichthyophonus prevalence and infection severity between upper Yukon River and Tanana River chinook salmon, Oncorhynchus tshawytscha (Walbaum), stocks

USGS Publications Warehouse

Kocan, R.; Hershberger, P.

2006-01-01

Two genetically distinct populations of chinook salmon, Oncorhynchus tshawytscha (Walbaum), were simultaneously sampled at the confluence of the Yukon and Tanana rivers in 2003. Upper Yukon-Canadian fish had significantly higher infection prevalence as well as more severe infections (higher parasite density in heart tissue) than the lower Yukon-Tanana River fish. Both populations had migrated the same distance from the mouth of the Yukon River at the time of sampling but had significantly different distances remaining to swim before reaching their respective spawning grounds. Multiple working hypotheses are proposed to explain the differences between the two stocks: (1) the two genetically distinct populations have different inherent resistance to infection, (2) genetically influenced differences in feeding behaviour resulted in temporal and/or spatial differences in exposure, (3) physiological differences resulting from different degrees of sexual maturity influenced the course of disease, and (4) the most severely infected Tanana River fish either died en route or fatigued and were unable to complete their migration to the Tanana River, thus leaving a population of apparently healthier fish. ?? 2006 Blackwell Publishing Ltd.

Interactive Video, The Next Step

NASA Astrophysics Data System (ADS)

Strong, L. R.; Wold-Brennon, R.; Cooper, S. K.; Brinkhuis, D.

2012-12-01

Video has the ingredients to reach us emotionally - with amazing images, enthusiastic interviews, music, and video game-like animations-- and it's emotion that motivates us to learn more about our new interest. However, watching video is usually passive. New web-based technology is expanding and enhancing the video experience, creating opportunities to use video with more direct interaction. This talk will look at an Educaton and Outreach team's experience producing video-centric curriculum using innovative interactive media tools from TED-Ed and FlixMaster. The Consortium for Ocean Leadership's Deep Earth Academy has partnered with the Center for Dark Energy Biosphere Investigations (C-DEBI) to send educators and a video producer aboard three deep sea research expeditions to the Juan de Fuca plate to install and service sub-seafloor observatories. This collaboration between teachers, students, scientists and media producers has proved a productive confluence, providing new ways of understanding both ground-breaking science and the process of science itself - by experimenting with new ways to use multimedia during ocean-going expeditions and developing curriculum and other projects post-cruise.

Neotectonic Activity from the Upper Reaches of the Arabian Gulf and Possibilities of New Oil Fields

NASA Astrophysics Data System (ADS)

Sissakian, V. K.; Abdul Ahad, A. D.; Al-Ansari, N.; Knutsson, S.

2018-03-01

Upper reaches of the Arabian Gulf consist of different types of fine sediments including the vast Mesopotamia Plain sediments, tidal flat sediments and estuarine sabkha sediments. The height of the plain starts from zero meter and increases northwards to three meters with extremely gentle gradient. The vast plain to the north of the Arabian Gulf is drained by Shat Al-Arab (Shat means river in Iraqi slang language) and Khor Al-Zubair (Khor means estuary). The former drains the extreme eastern part of the plain; whereas, the latter drains the western part. Shat Al-Arab is the resultant of confluence of the Tigris and Euphrates rivers near Al-Qurna town; about 160 km north of the Arabian Gulf mouth at Al-Fao town; whereas, the length of Khor Al-Zubair is about 50 km; as measured from Um Qasir Harbor. The drainage system around Khor Al-Zubair is extremely fine dendritic; whereas around Shat Al-Arab is almost parallel running from both sides of the river towards the river; almost perpendicularly. The fine dendritic drainage around Khor Al-Zubair shows clear recent erosional activity, beside water divides, abandoned irrigation channels and dislocated irrigational channels and estuarine distributaries; all are good indication for a Neotectonic activity in the region. These may indicate the presence of subsurface anticlines, which may represent oil fields; since tens of subsurface anticlines occur in near surroundings, which are oil fields.

The Propagation of a Surge Front on Bering Glacier, Alaska, 2001-2011

NASA Technical Reports Server (NTRS)

Turrin, James; Forster, Richard R.; Larsen, Chris; Sauber, Jeanne

2013-01-01

Bering Glacier, Alaska, USA, has a 20 year surge cycle, with its most recent surge reaching the terminus in 2011. To study this most recent activity a time series of ice velocity maps was produced by applying optical feature-tracking methods to Landsat-7 ETM+ imagery spanning 2001-11. The velocity maps show a yearly increase in ice surface velocity associated with the down-glacier movement of a surge front. In 2008/09 the maximum ice surface velocity was 1.5 plus or minus 0.017 kilometers per a in the mid-ablation zone, which decreased to 1.2 plus or minus 0.015 kilometers per a in 2009/10 in the lower ablation zone, and then increased to nearly 4.4 plus or minus 0.03 kilometers per a in summer 2011 when the surge front reached the glacier terminus. The surge front propagated down-glacier as a kinematic wave at an average rate of 4.4 plus or minus 2.0 kilometers per a between September 2002 and April 2009, then accelerated to 13.9 plus or minus 2.0 kilometers per a as it entered the piedmont lobe between April 2009 and September 2010. Thewave seems to have initiated near the confluence of Bering Glacier and Bagley Ice Valley as early as 2001, and the surge was triggered in 2008 further down-glacier in the mid-ablation zone after the wave passed an ice reservoir area.

Enhancing magnetic nanoparticle-based DNA transfection: Intracellular-active cassette features

NASA Astrophysics Data System (ADS)

Vernon, Matthew Martin

Efficient plasmid DNA transfection of embryonic stem cells, mesenchymal stem cells, neural cell lines and the majority of primary cell lines is a current challenge in gene therapy research. Magnetic nanoparticle-based DNA transfection is a gene vectoring technique that is promising because it is capable of outperforming most other non-viral transfection methods in terms of both transfection efficiency and cell viability. The nature of the DNA vector implemented depends on the target cell phenotype, where the particle surface chemistry and DNA binding/unbinding kinetics of the DNA carrier molecule play a critical role in the many steps required for successful gene transfection. Accordingly, Neuromag, an iron oxide/polymer nanoparticle optimized for transfection of neural phenotypes, outperforms many other nanoparticles and lipidbased DNA carriers. Up to now, improvements to nanomagnetic transfection techniques have focused mostly on particle functionalization and transfection parameter optimization (cell confluence, growth media, serum starvation, magnet oscillation parameters, etc.). None of these parameters are capable of assisting the nuclear translocation of delivered plasmid DNA once the particle-DNA complex is released from the endosome and dissociates in the cell's cytoplasm. In this study, incorporation of a DNA targeting sequence (DTS) feature in the transfecting plasmid DNA confers improved nuclear translocation, demonstrating significant improvement in nanomagnetic transfection efficiency in differentiated SH-SY5Y neuroblastoma cells. Other parameters, such as days in vitro, are also found to play a role and represent potential targets for further optimization.

JNK-associated scattered growth of YD-10B oral squamous carcinoma cells while maintaining the epithelial phenotype

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lee, Gayoung; Kim, Hyun-Man

Cell scattering of epithelial carcinoma cancer cells is one of the critical event in tumorigenesis. Cells losing epithelial cohesion detach from aggregated epithelial cell masses and may migrate to fatal organs through metastasis. The present study investigated the molecular mechanism by which squamous cell carcinoma cells grow scattered at the early phase of transformation while maintaining the epithelial phenotype. We studied YD-10B cells, which are established from human oral squamous cell carcinoma, because the cells grow scattered without the development of E-cadherin junctions (ECJs) under routine culture conditions despite the high expression of functional E-cadherin. The functionality of their E-cadherinmore » was demonstrated in that YD-10B cells developed ECJs, transiently or persistently, when they were cultured on substrates coated with a low amount of fibronectin or to confluence. The phosphorylation of JNK was up-regulated in YD-10B cells compared with that in human normal oral keratinocyte cells or human squamous cell carcinoma cells, which grew aggregated along with well-organized ECJs. The suppression of JNK activity induced the aggregated growth of YD-10B cells concomitant with the development of ECJs. These results indicate for the first time that inherently up-regulated JNK activity induces the scattered growth of the oral squamous cell carcinoma cells through down-regulating the development of ECJ despite the expression of functional E-cadherin, a hallmark of the epithelial phenotype. - Highlights: • JNK dissociates YD-10B oral squamous cell carcinoma cells. • JNK suppresses the development of E-cadherin junctions of oral carcinoma cells. • Suppression of JNK activity reverses the scattered growth of oral carcinoma cells.« less

Agent-Based Computational Modeling of Cell Culture ...

EPA Pesticide Factsheets

Quantitative characterization of cellular dose in vitro is needed for alignment of doses in vitro and in vivo. We used the agent-based software, CompuCell3D (CC3D), to provide a stochastic description of cell growth in culture. The model was configured so that isolated cells assumed a “fried egg shape” but became increasingly cuboidal with increasing confluency. The surface area presented by each cell to the overlying medium varies from cell-to-cell and is a determinant of diffusional flux of toxicant from the medium into the cell. Thus, dose varies among cells for a given concentration of toxicant in the medium. Computer code describing diffusion of H2O2 from medium into each cell and clearance of H2O2 was calibrated against H2O2 time-course data (25, 50, or 75 uM H2O2 for 60 min) obtained with the Amplex Red assay for the medium and the H2O2-sensitive fluorescent reporter, HyPer, for cytosol. Cellular H2O2 concentrations peaked at about 5 min and were near baseline by 10 min. The model predicted a skewed distribution of surface areas, with between cell variation usually 2 fold or less. Predicted variability in cellular dose was in rough agreement with the variation in the HyPer data. These results are preliminary, as the model was not calibrated to the morphology of a specific cell type. Future work will involve morphology model calibration against human bronchial epithelial (BEAS-2B) cells. Our results show, however, the potential of agent-based modeling

Topography and behavior of Sertoli cells in sparse culture during the transitional remodeling phase.

PubMed

Tung, P S; Choi, A H; Fritz, I B

1988-01-01

We report observations on the behavior of Sertoli cells in sparse culture during the period from the time of plating to the time of initial confluence (the transitional remodeling phase). Changes in shape, structure, and polarity of cells, as well as changes in migration patterns and cell-cell association patterns, have been followed during the transitional remodeling phase with the aid of topographical markers. These markers are based upon differences between ultrastructural features of the basolateral and apicolateral surfaces. The basolateral surface is characterized by plasmalemmal blebs, whereas the apicolateral surface is characterized by filopodial extensions. Structural differences observed in situ remain evident in Sertoli cells isolated by sequential enzymatic treatments that are described. Another marker is provided by laminin-binding sites, which are detected exclusively on the blebbed, basolateral surfaces of freshly prepared Sertoli cell aggregates. The orientation described is sustained during the initial radial migration of Sertoli cells explanted on uncoated glass coverslips. Under these conditions, blebs are detected only on the dorsal surfaces, and filopodial extensions are evident only on the ventral surfaces. In contrast, Sertoli cells sparsely plated on a reconstituted basement membrane (air-dried Matrigel) migrate rapidly, display an extraordinary capacity to form elaborate cytoplasmic extensions for cell-cell and cell-substratum contacts, and readily retract blebs and filopodial extensions. These cells do not form mosaic borders, whereas cells plated on uncoated glass do form a monolayer with mosaic-like borders. Cells sparsely seeded on gelated Matrigel migrate preferentially at gaps between adjacent cell explants, and develop a compact cell-cell association pattern. These cells display few, if any, cytoplasmic extensions. We compare the behavior of Sertoli cells sparsely plated on Matrigel with the behavior of Sertoli cells in situ during different stages of development.

Ecological risk assessment of heavy metals and polycyclic aromatic hydrocarbons in sediments of rivers Niger and Benue confluence, Lokoja, Central Nigeria.

PubMed

Ekere, Nwachukwu; Yakubu, Newman; Ihedioha, Janefrances

2017-08-01

The concentrations of six heavy metals (HMs) and 16 US EPA priority polycyclic aromatic hydrocarbons (PAHs) in sediment samples of the confluence of rivers Niger and Benue were investigated. The ecological risk assessment of the contaminants was carried out. The results showed that the sediment samples were heavily polluted with iron and moderately polluted with Cd while other metals posed no pollution problem when compared with USEPA sediment quality guidelines. Only six out of the 16 priority PAHs were detected in the samples, and source apportionment of the PAHs indicated that they are of pyrogenic origin. The ∑PAHs in the samples were lower than many of similar studies and were of no pollution risk. The ecological risk assessment result of the heavy metals showed that the sediments were of considerable risk due majorly to Cd levels. The HM concentration results statistically showed significant difference between seasons at probability value (P < .05). Data analysis by PCA classified the metals into three different components according to sources. The levels of HMS and PAHs detected in the sediments were correlated for source identification, and the correlation showed that the majority of the pollutants were mainly from anthropogenic sources. There is increasing level of anthropogenic activities at the vicinity of the confluence due to urbanization which may call for periodic monitoring of the sediment quality.

A biologically based model of growth and senescence of Syrian hamster embryo (SHE) cells after exposure to arsenic.

PubMed Central

Liao, K H; Gustafson, D L; Fox, M H; Chubb, L S; Reardon, K F; Yang, R S

2001-01-01

We modified the two-stage Moolgavkar-Venzon-Knudson (MVK) model for use with Syrian hamster embryo (SHE) cell neoplastic progression. Five phenotypic stages are proposed in this model: Normal cells can either become senescent or mutate into immortal cells followed by anchorage-independent growth and tumorigenic stages. The growth of normal SHE cells was controlled by their division, death, and senescence rates, and all senescent cells were converted from normal cells. In this report, we tested the modeling of cell kinetics of the first two phenotypic stages against experimental data evaluating the effects of arsenic on SHE cells. We assessed cell division and death rates using flow cytometry and correlated cell division rates to the degree of confluence of cell cultures. The mean cell death rate was approximately equal to 1% of the average division rate. Arsenic did not induce immortalization or further mutations of SHE cells at concentrations of 2 microM and below, and chromium (3.6 microM) and lead (100 microM) had similar negative results. However, the growth of SHE cells was inhibited by 5.4 microM arsenic after a 2-day exposure, with cells becoming senescent after only 16 population doublings. In contrast, normal cells and cells exposed to lower arsenic concentrations grew normally for at least 30 population doublings. The biologically based model successfully predicted the growth of normal and arsenic-treated cells, as well as the senescence rates. Mechanisms responsible for inducing cellular senescence in SHE cells exposed to arsenic may help explain the apparent inability of arsenic to induce neoplasia in experimental animals. PMID:11748027

Potential Role of S100A8 in Cutaneous Squamous Cell Carcinoma Differentiation

PubMed Central

Shin, Jung-Min; Chang, In-Kyu; Lee, Young-Ho; Yeo, Min-Kyung; Kim, Jin-Man; Sohn, Kyung-Cheol; Im, Myung; Seo, Young-Joon; Kim, Chang-Deok; Lee, Jeung-Hoon

2016-01-01

Background S100A8 is differentially expressed in various cell types and is associated with a number of malignant disorders. S100A8 may affect tumor biology. However, its role in cutaneous squamous cell carcinoma (SCC) is not well established. Objective This study aims to investigate the relationship between S100A8 and cutaneous SCC development. Methods We performed immunohistochemical staining to detect S100A8 expression in facial skin specimens of premalignant actinic keratosis (AK), malignant SCC, and normal tissues. In addition, we utilized postconfluence and high calcium-induced differentiation in a culture system model. Furthermore, we constructed a recombinant adenovirus expressing GFP-tagged S100A8 to investigate the role of S100A8 in SCC cell differentiation. Results S100A8 was significantly overexpressed in human cutaneous SCC compared to that in normal and AK tissues. S100A8 was gradually upregulated in SCC cells in a post-confluence-induced differentiation model. Overexpression of S100A8 in SCC cells induced by adenoviral transduction led to increased expression levels of differentiation markers, such as loricrin, involucrin, and filaggrin. S100A8 overexpression also increased loricrin and involucrin luciferase activity. Conclusion S100A8 regulates cutaneous SCC differentiation and induces well-differentiated SCC formation in skin. PMID:27081264

EFFECTS OF PLATING DENSITY AND CULTURE TIME ON BONE MARROW STROMAL CELL CHARACTERISTICS

PubMed Central

Neuhuber, Birgit; Swanger, Sharon A.; Howard, Linda; Mackay, Alastair; Fischer, Itzhak

2008-01-01

Objective Bone marrow stromal cells (MSC) are multipotent adult stem cells that have emerged as promising candidates for cell therapy in disorders including cardiac infarction, stroke and spinal cord injury. While harvesting methods used by different laboratories are relatively standard, MSC culturing protocols vary widely. This study is aimed at evaluating the effects of initial plating density and total time in culture on proliferation, cell morphology, and differentiation potential of heterogeneous MSC cultures and more homogeneous cloned subpopulations. Methods Rat MSC were plated at 20, 200 and 2000 cells/cm2 and grown to 50% confluency. The numbers of population doublings and doubling times were determined within and across multiple passages. Changes in cell morphology and differentiation potential to adipogenic, chondrogenic, and osteogenic lineages were evaluated and compared among early, intermediate and late passages, as well as between heterogeneous and cloned MSC populations. Results We found optimal cell growth at a plating density of 200 cells/cm2. Cultures derived from all plating densities developed increased proportions of flat cells over time. Assays for chondrogenesis, osteogenesis and adipogenesis showed that heterogeneous MSC plated at all densities sustained the potential for all three mesenchymal phenotypes through at least passage 5; the flat subpopulation lost adipogenic and chondrogenic potential. Conclusion Our findings suggest that the initial plating density is not critical for maintaining a well-defined, multipotent MSC population. Time in culture, however, affects cell characteristics, suggesting that cell expansion should be limited, especially until the specific characteristics of different MSC subpopulations are better understood. PMID:18495329

Drainage areas of the Potomac River basin, West Virginia

USGS Publications Warehouse

Wiley, Jeffrey B.; Hunt, Michelle L.; Stewart, Donald K.

1996-01-01

This report contains data for 776 drainage-area divisions of the Potomac River Basin, from the headwaters to the confluence of the Potomac River and the Shenandoah River. Data, compiled in downstream order, are listed for streams with a drainage area of approximately 2 square miles or larger within West Virginia and for U.S. Geological Survey streamflow-gaging stations. The data presented are the stream name, the geographical limits in river miles, the latitude and longitude of the point, the name of the county, and the 7 1/2-minute quadrangle in which the point lies, and the drainage area of that site. The total drainage area of the Potomac River Basin downstream of the confluence of the Shenandoah River at the State boundary is 9,367.29 square miles.

The fate of arsenic in sediments formed at a river confluence affected by acid mine drainage

NASA Astrophysics Data System (ADS)

Guerra, P. A.; Pasten, P. A.; Pizarro, G.; Simonson, K.; Escauriaza, C. R.; Gonzalez, C.; Bonilla, C.

2012-12-01

Fluvial confluences receiving acid mine drainage may play a critical role in a watershed as a suite of interactions between chemistry and hydrodynamics occur, determining the fate of toxic contaminants like arsenic. Solid reactive phases of iron and/or aluminum oxi-hydroxides may form or transform, ranging from iron oxide nanoparticles that aggregate and form floccules that are transported in the suspended load up to gravel and arsenic-rich rock coatings. In order to further understand the role of reactive fluvial confluences, we have studied the mixing between the Caracarani River (flow=170-640 L/s, pH 8, conductivity 1.5 mS/cm, total As<0.1 mg/L and total Fe< 5 mg/L) and the Azufre River (flow=45-245 L/s, pH<2, conductivity > 10 mS/cm, total As>2 mg/L, total Fe=35-125 mg/L), located in the Lluta watershed in northern Chile. This site is an excellent natural laboratory located in a water-scarce area, where the future construction of a dam has prompted the attention of decision makers and scientists interested in weighing the risks derived by the accumulation of arsenic-rich sediments. Suspended sediments (> 0.45 μm), riverbed sediments, and coated rocks were collected upstream and downstream from the confluence. Suspended sediments >0.45 μm and riverbed sediments were analyzed by total reflection x-ray fluorescence for metals, while coated river bed rocks were analyzed by chemical extractions and a semi-quantitative approach through portable x-ray fluorescence. Water from the Caracarani and Azufre rivers were mixed in the laboratory at different ratios and mixing velocities aiming to characterize the effect of the chemical-hydrodynamic environment where arsenic solids were formed at different locations in the confluence. Despite a wide range of iron and arsenic concentrations in the suspended sediments from the field (As=1037 ± 1372 mg/kg, Fe=21.0 ± 24.5 g/kg), we found a rather narrow As/Fe ratio, increasing from 36.5 to 55.2 mgAs/kgFe when the bulk water pH increased from 3 to 6. Sequential extraction analyses suggest that ~80% of As in the solid pahse is strongly sorbed to the sediments, whereas ~15% of As is forming co-precipitated phase. Riverbed sediments (sand and gravel) showed much lower concentrations of Fe and As (17.1 ± 3.0 g/kg and 67.5 ± 53.9 mg/kg, respectively), owing to a dilution from non-reactive phases like quartz from sand. Coated rocks showed concentrations of Fe and As of 34.8 ± 12.5 g/kg and 680 ± 401 mg/kg, respectively, suggesting the coatings are possibly constituted by As-rich particles sequestered from the flow. In the case of the laboratory mixing tests, suspended solids ranged from 10 to 60 mg/L, with higher values at lower mixing ratios or higher pHs. Our findings suggest that the spatial and temporal variability driven by the hydrodynamics of a confluence determine distinct geochemical characteristics of arsenic-rich solid phases, thus playing a role in the fate of reactive contaminants like arsenic in the watershed. Research funded by Fondecyt project 1100943.

Gas6 Induces Growth, β-Catenin Stabilization, and T-Cell Factor Transcriptional Activation in Contact-Inhibited C57 Mammary Cells

PubMed Central

Goruppi, Sandro; Chiaruttini, Cristina; Ruaro, Maria Elisabetta; Varnum, Brian; Schneider, Claudio

2001-01-01

Gas6 is a growth factor related to protein S that was identified as the ligand for the Axl receptor tyrosine kinase (RTK) family. In this study, we show that Gas6 induces a growth response in a cultured mammalian mammary cell line, C57MG. The presence of Gas6 in the medium induces growth after confluence and similarly causes cell cycle reentry of density-inhibited C57MG cells. We show that Axl RTK but not Rse is efficiently activated by Gas6 in density-inhibited C57MG cells. We have analyzed the signaling required for the Gas6 proliferative effect and found a requirement for PI3K-, S6K-, and Ras-activated pathways. We also demonstrate that Gas6 activates Akt and concomitantly inhibits GSK3 activity in a wortmannin-dependent manner. Interestingly, Gas6 induces up-regulation of cytosolic β-catenin, while membrane-associated β-catenin remains unaffected. Stabilization of β-catenin in C57MG cells is correlated with activation of a T-cell factor (TCF)-responsive transcriptional element. We thus provide evidence that Gas6 is mitogenic and induces β-catenin proto-oncogene stabilization and subsequent TCF/Lef transcriptional activation in a mammary system. These results suggest that Gas6-Axl interaction, through stabilization of β-catenin, may have a role in mammary development and/or be involved in the progression of mammary tumors. PMID:11154277

A Method for the Immortalization of Newborn Mouse Skin Keratinocytes

PubMed Central

Hammiller, Brianna O.; El-Abaseri, Taghrid Bahig; Dlugosz, Andrzej A.; Hansen, Laura A.

2015-01-01

Isolation and culture of mouse primary epidermal keratinocytes is a common technique that allows for easy genetic and environmental manipulation. However, due to their limited lifespan in culture, experiments utilizing primary keratinocytes require large numbers of animals, and are time consuming and expensive. To avoid these issues, we developed a method for the immortalization of primary mouse epidermal keratinocytes. Upon isolation of newborn epidermal keratinocytes according to established methods, the cells were cultured long-term in keratinocyte growth factor-containing medium. The cells senesced within a few weeks and eventually, small, slowly growing colonies emerged. After they regained confluency, the cells were passaged and slowly refilled the dish. With several rounds of subculture, the cells adapted to culture conditions, were easily subcultured, maintained normal morphology, and were apparently immortal. The immortalized cells retained the ability to differentiate with increased calcium concentrations, and were maintained to high passage numbers while maintaining a relatively stable karyotype. Analysis of multiple immortalized cell lines as well as primary keratinocyte cultures revealed increased numbers of chromosomes, especially in the primary keratinocytes, and chromosomal aberrations in most of the immortalized cultures and in the primary keratinocytes. Orthotopic grafting of immortalized keratinocytes together with fibroblasts onto nude mouse hosts produced skin while v-rasHa infection of the immortalized keratinocytes prior to grafting produced squamous cell carcinoma. In summary, this method of cell line generation allows for decreased use of animals, reduces the expense and time involved in research, and provides a useful model for cutaneous keratinocyte experimentation. PMID:26284198

75 FR 59192 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-27

... of Perry, downstream of the Township of Toby. confluence with Black Fox Run. Approximately 480 feet... No. 97.022, ``Flood Insurance.'') Dated: September 13, 2010. Sandra K. Knight, Deputy Federal...

Polyelectrolyte Multilayer-Treated Electrodes for Real-Time Electronic Sensing of Cell Proliferation

PubMed Central

Mijares, Geraldine I.; Reyes, Darwin R.; Geist, Jon; Gaitan, Michael; Polk, Brian J.; DeVoe, Don L.

2010-01-01

We report on the use of polyelectrolyte multilayer (PEM) coatings as a non-biological surface preparation to facilitate uniform cell attachment and growth on patterned thin-film gold (Au) electrodes on glass for impedance-based measurements. Extracellular matrix (ECM) proteins are commonly utilized as cell adhesion promoters for electrodes; however, they exhibit degradation over time, thereby imposing limitations on the duration of conductance-based biosensor experiments. The motivation for the use of PEM coatings arises from their long-term surface stability as promoters for cell attachment, patterning, and culture. In this work, a cell proliferation monitoring device was fabricated. It consisted of thin-film Au electrodes deposited with a titanium-tungsten (TiW) adhesion layer that were patterned on a glass substrate and passivated to create active electrode areas. The electrode surfaces were then treated with a poly(ethyleneimine) (PEI) anchoring layer and subsequent bilayers of sodium poly(styrene sulfonate) (PSS) and poly(allylamine hydrochloride) (PAH). NIH-3T3 mouse embryonic fibroblast cells were cultured on the device, observed by optical microscopy, and showed uniform growth characteristics similar to those observed on a traditional polystyrene cell culture dish. The optical observations were correlated to electrical measurements on the PEM-treated electrodes, which exhibited a rise in impedance with cell proliferation and stabilized to an approximate 15 % increase as the culture approached confluency. In conclusion, cells proliferate uniformly over gold and glass PEM-treated surfaces, making them useful for continuous impedance-based, real-time monitoring of cell proliferation and for the determination of cell growth rate in cellular assays. PMID:27134780

Combined use of stable isotopes and hydrologic modeling to better understand nutrient sources and cycling in highly altered systems (Invited)

NASA Astrophysics Data System (ADS)

Young, M. B.; Kendall, C.; Guerin, M.; Stringfellow, W. T.; Silva, S. R.; Harter, T.; Parker, A.

2013-12-01

The Sacramento and San Joaquin Rivers provide the majority of freshwater for the San Francisco Bay Delta. Both rivers are important sources of drinking and irrigation water for California, and play critical roles in the health of California fisheries. Understanding the factors controlling water quality and primary productivity in these rivers and the Delta is essential for making sound economic and environmental water management decisions. However, these highly altered surface water systems present many challenges for water quality monitoring studies due to factors such as multiple potential nutrient and contaminant inputs, dynamic source water inputs, and changing flow regimes controlled by both natural and engineered conditions. The watersheds for both rivers contain areas of intensive agriculture along with many other land uses, and the Sacramento River receives significant amounts of treated wastewater from the large population around the City of Sacramento. We have used a multi-isotope approach combined with mass balance and hydrodynamic modeling in order to better understand the dominant nutrient sources for each of these rivers, and to track nutrient sources and cycling within the complex Delta region around the confluence of the rivers. High nitrate concentrations within the San Joaquin River fuel summer algal blooms, contributing to low dissolved oxygen conditions. High δ15N-NO3 values combined with the high nitrate concentrations suggest that animal manure is a significant source of nitrate to the San Joaquin River. In contrast, the Sacramento River has lower nitrate concentrations but elevated ammonium concentrations from wastewater discharge. Downstream nitrification of the ammonium can be clearly traced using δ15N-NH4. Flow conditions for these rivers and the Delta have strong seasonal and inter-annual variations, resulting in significant changes in nutrient delivery and cycling. Isotopic measurements and estimates of source water contributions derived from the DSM2-HYDRO hydrologic model demonstrate that mixing between San Joaquin and Sacramento River water can occur as far as 30 miles upstream of the confluence within the San Joaquin channel, and that San Joaquin-derived nitrate only reaches the western Delta during periods of high flow.

Interactions between Point Bar Growth and Bank Erosion on a Low Sinuosity Meander Bend in an Ephemeral Channel: Insights from Repeat Topographic Surveys and Numerical Modeling

NASA Astrophysics Data System (ADS)

Ursic, M.; Langendoen, E. J.

2017-12-01

Interactions between point bar growth, bank migration, and hydraulics on meandering rivers are complicated and not well understood. For ephemeral streams, rapid fluctuations in flow further complicate studying and understanding these interactions. This study seeks to answer the following `cause-and-effect' question: Does point bar morphologic adjustment determine where bank erosion occurs (for example, through topographic steering of the flow), or does local bank retreat determine where accretion/erosion occurs on the point bar, or do bank erosion and point bar morphologic adjustment co-evolve? Further, is there a response time between the `cause-and-effect' processes and what variables determine its magnitude and duration? In an effort to answer these questions for an ephemeral stream, a dataset of forty-eight repeat topographic surveys over a ten-year period (1996-2006) of a low sinuosity bend within the Goodwin Creek Experimental Watershed, located near Batesville, MS, were utilized in conjunction with continuous discharge measurements to correlate flow variability and erosional and depositional zones, spatially and temporally. Hydraulically, the bend is located immediately downstream of a confluence with a major tributary. Supercritical flumes on both the primary and tributary channels just upstream of the confluence provide continuous measured discharges to the bend over the survey period. In addition, water surface elevations were continuously measured at the upstream and downstream ends of the bend. No spatial correlation trends could be discerned between reach-scale bank retreat, point bar morphologic adjustment, and flow discharge. Because detailed flow patterns were not available, the two-dimensional computer model Telemac2D was used to provide these details. The model was calibrated and validated for a set of runoff events for which more detailed flow data were available. Telemac2D simulations were created for each topographic survey period. Flows greater than baseflow were combined to create contiguous hydrographs for each survey period. Statistical examination of local flow variability and morphological changes throughout the bend will be conducted and presented.

Comparative evaluation of the isolation and quantification of stem cells derived from dental pulp and periodontal ligament of a permanent tooth and to assess their viability and proliferation on a platelet-rich fibrin scaffold.

PubMed

Khurana, Rohit; Kudva, Praveen Bhasker; Husain, Syed Yawer

2017-01-01

The present study aims to comparatively evaluate the isolation and quantification of stem cells derived from dental pulp and periodontal ligament of a permanent tooth and to assess their viability and proliferation on a platelet-rich fibrin (PRF) scaffold. A total of 15 systemically healthy individuals between the age group of 15-25 years requiring third molar or orthodontic premolar extractions. Teeth were extracted atraumatically and transported to the laboratory. Stem cells were isolated from dental pulp and periodontal ligament. After attaining more than 90% confluency by the 7 th day, these cells were tested for their viability and characterization. Stem cells were also incubated with PRF and viability was assessed on the 7 th day. The mean number of cell for dental pulp stem cells (DPSCs) and periodontal ligament stem cell (PDLSC) was statistically insignificant ( P > 0.05). The mean live cell viability was compared between DPSC (98.07%) and PDLSC (98%). Both DPSC and PDLSC showed a high percentage of expression of CD73 markers, 30.40% and 29.80%, respectively. However, DPSCs and PDLSCs lacked expression of CD34 expressing only 3.47% and 3.53%, respectively. PRF membrane as a scaffold exhibited no cytotoxic effects on DPCS's or PDLSC's. The cell viability of cells cultured with PRF was statistically insignificant ( P > 0.05) when compared to the cells cultured with culture media. The study thus indicates that dental pulp and periodontal ligament are both rich sources of mesenchymal stem cells and can be successfully used for obtaining stem cells. PRF exhibits no cytotoxic effects on the cells and can be used in conjunction with dental stem cells.

76 FR 72627 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-25

.... confluence with Black Fox Run. Approximately 480 feet +868 upstream of State Route 368. Allegheny River... Assistance No. 97.022, ``Flood Insurance.'' Dated: November 14, 2011. Sandra K. Knight, Deputy Associate...

77 FR 49367 - Final Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2012-08-16

... County. Verde Creek confluence. Approximately 500 feet +2593 downstream of the Agua Caliente Wash... Pima County. Tanque Verde Road. Approximately 0.4 mile +2624 upstream of East Tanque Verde Road. Agua...

Chickamauga National Military Park Tour Roads, Gordon's Slough Bridge, At ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Chickamauga National Military Park Tour Roads, Gordon's Slough Bridge, At the confluence of Alexander's Bridge Road and Gordon's Slough, southeast of Alexander's Bridge, Fort Oglethorpe, Catoosa County, GA

Hydrodynamics of concordant and discordant fixed bed open-channel confluences

NASA Astrophysics Data System (ADS)

Birjukova Canelas, Olga; Lage Ferreira, Rui Miguel; Heleno Cardoso, António

2017-04-01

The detailed characterization of the flow field in river confluences constitutes a relevant step towards the understanding of the hydro-morpho-dynamics of these key zones of the fluvial system. With a few exceptions, existing works on this topic covered concordant bed scenarios, meaning that both confluent channels had the same elevation. This laboratory study aims to contribute to a detailed three-dimensional characterization of the flow field at a fixed bed confluence, as well as to shed light on how bed elevation discordance modifies the flow patterns of the converging flows. While the junction angle and the discharge ratio were kept fixed, two scenarios were studied on the basis of detailed water level and 3D ADV measurements at the denser mesh ever. The internal flow structure of the concordant bed scenario mostly complied with the classical conceptual models. A relevant difference concerns the size of the stagnation zone, much smaller close to the bed of the discordant bed confluence. A more significant difference is a horizontal flow structure, not previously identified in the literature, characterized by strong streamwise mean vorticity and strong secondary motion. It is observed for the discordant bed case, occurring along the inner wall of the main channel and downstream the junction corner. This structure is spatially well-correlated to a pronounced imbalance of cross-stream and vertical normal Reynolds stresses. This highlights the role of Reynolds stress anisotropy (RSA) that is generated in the shear layers than accompany the entrance of the tributary flow. Since this structure is not present in the concordant case, where RSA is also evident, it is argued that convective effects should also play a role in its formation, presumably due to deflection of the flow in the main channel by the tributary. The newly identified secondary motion should, thus, be a combination of Prandtĺs second kind and Prandtĺs first kind of secondary flow. The relative importance of each generating mechanism is still under investigation. Acknowledgements This research as partially supported by Portuguese and European funds, within programs COMPETE2020 and PORL-FEDER, through project PTDC/ECM-HID/6387/2014 granted by the National Foundation for Science and Technology (FCT).

Further thoughts about "nothing".

PubMed

Shengold, L

1989-04-01

Some meanings of "nothing" are delineated according to their acquisition, confluence, and transformation in the course of the child's development of drives, the ego, and object relations. This sketch is illustrated mainly from King Lear.

Zinc and dexamethasone induce metallothionein accumulation by endothelial cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Briske-Anderson, M.; Bobilya, D.J.; Reeves, P.G.

1991-03-11

Several tissues increase their metallothionein (MT) concentration when exposed to elevated amounts of plasma Zn. Endothelial cells form the blood vessels that supply all tissues and constitute a barrier between cells of tissues and the blood. This study examined the ability of endothelial cells to synthesize MT and accumulate Zn in response to high amounts of Zn and dexamethasone. Bovine pulmonary endothelial cells were grown to confluence in Minimum Essential Medium with Earle's salts and 10% fetal calf serum. The monolayer was maintained for 2 d prior to use in medium containing EDTA-dialyzed serum. This low Zn medium was replacedmore » with one containing 1, 6, 25, 50, 100, 150, or 200 {mu}M Zn and incubated for 24 hr before harvesting the cells. MT was quantified by the cadmium binding assay. Cellular Zn concentrations were analyzed by atomic absorption after a nitric acid digestion. The MT concentration was elevated in response to Zn concentrations of 100 {mu}M or more. Cellular Zn concentration was elevated when media Zn was 25 {mu}M or more. MT and cellular Zn concentrations were positively correlated. In another study, inclusion of 0.1 {mu}M dexamethasone in the media increased concentration at all Zn concentrations studied. However, the inclusion of 0.3 {mu}M cis-platinum had no effect. In conclusion, endothelial cells in culture respond to elevated amounts of Zn and dexamethasone in the media by accumulating Zn and MT.« less

Effect of Stratification on Surface Properties of Corneal Epithelial Cells

PubMed Central

Yáñez-Soto, Bernardo; Leonard, Brian C.; Raghunathan, Vijay Krishna; Abbott, Nicholas L.; Murphy, Christopher J.

2015-01-01

Purpose The purpose of this study was to determine the influence of mucin expression in an immortalized human corneal epithelial cell line (hTCEpi) on the surface properties of cells, such as wettability, contact angle, and surface heterogeneity. Methods hTCEpi cells were cultured to confluence in serum-free medium. The medium was then replaced by stratification medium to induce mucin biosynthesis. The mucin expression profile was analyzed using quantitative PCR and Western blotting. Contact angles were measured using a two-immiscible liquid method, and contact angle hysteresis was evaluated by tilting the apparatus and recording advancing and receding contact angles. The spatial distribution of mucins was evaluated with fluorescently labeled lectin. Results hTCEpi cells expressed the three main ocular mucins (MUC1, MUC4, and MUC16) with a maximum between days 1 and 3 of the stratification process. Upon stratification, cells caused a very significant increase in contact angle hysteresis, suggesting the development of spatially discrete and heterogeneously distributed surface features, defined by topography and/or chemical functionality. Although atomic force microscopy measurements showed no formation of appreciable topographic features on the surface of the cells, we observed a significant increase in surface chemical heterogeneity. Conclusions The surface chemical heterogeneity of the corneal epithelium may influence the dynamic behavior of tear film by “pinning” the contact line between the cellular surface and aqueous tear film. Engineering the surface properties of corneal epithelium could potentially lead to novel treatments in dry eye disease. PMID:26747762

Hydraulic conductivity of endothelial cell-initiated arterial cocultures.

PubMed

Mathura, Rishi A; Russell-Puleri, Sparkle; Cancel, Limary M; Tarbell, John M

2014-04-01

This study describes cocultures of arterial smooth muscle cells (SMCs) and endothelial cells (ECs) and the influences of their heterotypic interactions on hydraulic conductivity (L p ), an important transport property. A unique feature of these cocultures is that ECs were first grown to confluence and then SMCs were inoculated. Bovine aortic smooth muscle cells and bovine aortic endothelial cells (BAECs) were cocultured on Transwell Permeable Supports, and then exposed to a pressure-driven transmural flow. L p across each culture was measured using a bubble tracking apparatus that determined water flux (J v ). Our results indicate that arterial L p is significantly modulated by EC-SMC proximity, and serum content in culture. The L p of cocultures was also compared to the predictions of a resistances-in-series model to distinguish the contributions of heterotypic interactions between SMCs and ECs. Conditions that lead to significantly reduced coculture L p , compared to BAEC monoculture controls, have been uncovered and the lowest L p in the literature for an in vitro system are reported. In addition, VE-cadherin immunostaining of intact BAEC monolayers in each culture configuration reveals that EC-SMC proximity on a porous membrane has a dramatic influence on EC morphology patterns. The cocultures with the lowest L p have ECs with significantly elongated morphology. Confocal imaging indicates that there are no direct EC-SMC contacts in coculture.

Medium Calcium Concentration Determines Keratin Intermediate Filament Density and Distribution in Immortalized Cultured Thymic Epithelial Cells (TECs)

NASA Astrophysics Data System (ADS)

Sands, Sandra S.; Meek, William D.; Hayashi, Jun; Ketchum, Robert J.

2005-08-01

Isolation and culture of thymic epithelial cells (TECs) using conventional primary tissue culture techniques under conditions employing supplemented low calcium medium yielded an immortalized cell line derived from the LDA rat (Lewis [Rt1l] cross DA [Rt1a]) that could be manipulated in vitro. Thymi were harvested from 4 5-day-old neonates, enzymically digested using collagenase (1 mg/ml, 37°C, 1 h) and cultured in low calcium WAJC404A medium containing cholera toxin (20 ng/ml), dexamethasone (10 nM), epidermal growth factor (10 ng/ml), insulin (10 [mu]g/ml), transferrin (10 [mu]g/ml), 2% calf serum, 2.5% Dulbecco's Modified Eagle's Medium (DMEM), and 1% antibiotic/antimycotic. TECs cultured in low calcium displayed round to spindle-shaped morphology, distinct intercellular spaces (even at confluence), and dense reticular-like keratin patterns. In high calcium (0.188 mM), TECs formed cobblestone-like confluent monolayers that were resistant to trypsinization (0.05%) and displayed keratin intermediate filaments concentrated at desmosomal junctions between contiguous cells. Changes in cultured TEC morphology were quantified by an analysis of desmosome/membrane relationships in high and low calcium media. Desmosomes were significantly increased in the high calcium medium. These studies may have value when considering the growth conditions of cultured primary cell lines like TECs.

Biocompatibility of nanostructured boron doped diamond for the attachment and proliferation of human neural stem cells.

PubMed

Taylor, Alice C; Vagaska, Barbora; Edgington, Robert; Hébert, Clément; Ferretti, Patrizia; Bergonzo, Philippe; Jackman, Richard B

2015-12-01

We quantitatively investigate the biocompatibility of chemical vapour deposited (CVD) nanocrystalline diamond (NCD) after the inclusion of boron, with and without nanostructuring. The nanostructuring method involves a novel approach of growing NCD over carbon nanotubes (CNTs) that act as a 3D scaffold. This nanostructuring of BNCD leads to a material with increased capacitance, and this along with wide electrochemical window makes BNCD an ideal material for neural interface applications, and thus it is essential that their biocompatibility is investigated. Biocompatibility was assessed by observing the interaction of human neural stem cells (hNSCs) with a variety of NCD substrates including un-doped ones, and NCD doped with boron, which are both planar, and nanostructured. hNSCs were chosen due to their sensitivity, and various methods including cell population and confluency were used to quantify biocompatibility. Boron inclusion into NCD film was shown to have no observable effect on hNSC attachment, proliferation and viability. Furthermore, the biocompatibility of nanostructured boron-doped NCD is increased upon nanostructuring, potentially due to the increased surface area. Diamond is an attractive material for supporting the attachment and development of cells as it can show exceptional biocompatibility. When boron is used as a dopant within diamond it becomes a p-type semiconductor, and at high concentrations the diamond becomes quasi-metallic, offering the prospect of a direct electrical device-cell interfacing system.

2. VIEW OF POWER PLANT LOOKING SOUTHEAST. Potomac Power ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

2. VIEW OF POWER PLANT LOOKING SOUTHEAST. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

Surgical Treatment for Biliary Carcinoma Arising After Pancreatoduodenectomy

PubMed Central

Seki, Hitoshi; Kobayashi, Akira; Kawasaki, Seiji

1998-01-01

The clinicopathological features and surgical treatment of biliary carcinoma around the major hepatic duct confluence arising after pancreatoduodenectomy (PD) due to initial bile duct carcinoma are described in three patients. Occurrence of biliary carcinoma more than 12 years after initial surgery and a histological finding of cholangiocellular carcinoma mixed with hepatocellular carcinoma suggested metachronous incidence of biliary carcinoma after PD. Extended right hemihepatectomy with complete removal of the residual extrahepatic bile duct and segmental, resection of the jejunal loop were carried out safely without operative death or severe postoperative complications. Two patients died of tumor recurrence 6 months after surgery, and the remaining patient is currently living a normal life without evidence of recurrence 17 months after surgery. These surgical procedures are a therapeutic option in patients with biliary carcinoma around the major hepatic duct confluence arising after PD. PMID:9515238

Imaging the equilibrium state and magnetization dynamics of partially built hard disk write heads

DOE Office of Scientific and Technical Information (OSTI.GOV)

Valkass, R. A. J., E-mail: rajv202@ex.ac.uk; Yu, W.; Shelford, L. R.

Four different designs of partially built hard disk write heads with a yoke comprising four repeats of NiFe (1 nm)/CoFe (50 nm) were studied by both x-ray photoemission electron microscopy (XPEEM) and time-resolved scanning Kerr microscopy (TRSKM). These techniques were used to investigate the static equilibrium domain configuration and the magnetodynamic response across the entire structure, respectively. Simulations and previous TRSKM studies have made proposals for the equilibrium domain configuration of similar structures, but no direct observation of the equilibrium state of the writers has yet been made. In this study, static XPEEM images of the equilibrium state of writer structures weremore » acquired using x-ray magnetic circular dichroism as the contrast mechanism. These images suggest that the crystalline anisotropy dominates the equilibrium state domain configuration, but competition with shape anisotropy ultimately determines the stability of the equilibrium state. Dynamic TRSKM images were acquired from nominally identical devices. These images suggest that a longer confluence region may hinder flux conduction from the yoke into the pole tip: the shorter confluence region exhibits clear flux beaming along the symmetry axis, whereas the longer confluence region causes flux to conduct along one edge of the writer. The observed variations in dynamic response agree well with the differences in the equilibrium magnetization configuration visible in the XPEEM images, confirming that minor variations in the geometric design of the writer structure can have significant effects on the process of flux beaming.« less

Linking hydro-morphology with invertebrate ecology in diverse morphological units of a large river-floodplain system

NASA Astrophysics Data System (ADS)

Blettler, Martín. C. M.; Amsler, Mario L.; Eberle, Eliana G.; Szupiany, Ricardo; Latosinski, Francisco G.; Abrial, Elie; Oberholster, Paul J.; Espinola, Luis A.; Paira, Aldo; Poza, Ailen; Rodrigues Capítulo, Alberto

2016-12-01

Interdisciplinary research in the fields of ecohydrology and ecogeomorphology is becoming increasingly important as a way to understand how biological and physical processes interact with each other in river systems. The objectives of the current study were 1) to determine changes in invertebrate community due to hydrological stages, 2) to link local physical features [flow configuration, sediment composition and morphological feature) with the ecological structure between and within dissimilar morphological units (meander and confluence), and 3) to determine the existence and the origin of bed hydro-geomorphic patches, determining their ecological structure. Results were discussed in the frame of prevailing ecological models and concepts. The study site extends over a floodplain area of the large Paraná River (Argentina), including minor and major secondary channels as well as the main channel. Overall results suggested that hydrodynamics was the driving force determining distribution patterns of benthic assemblages in the floodplain. However, while the invertebrates living in minor secondary channels seem to benefit from flooding, this hydrological phase had the opposite effect on organisms from the main and major secondary channels. We also found a clear linkage between physical features and invertebrate ecology, which caused a dissimilar fauna structure between and within the meander and the confluence. Furthermore, several sandy-patches were recorded in the confluence. These patches were colonized by the particular benthic assemblage recorded in the main channel, supported the view of rivers as patchy discontinua, under uncertain ecological equilibrium.

Growth and differentiation of human lens epithelial cells in vitro on matrix

NASA Technical Reports Server (NTRS)

Blakely, E. A.; Bjornstad, K. A.; Chang, P. Y.; McNamara, M. P.; Chang, E.; Aragon, G.; Lin, S. P.; Lui, G.; Polansky, J. R.

2000-01-01

PURPOSE: To characterize the growth and maturation of nonimmortalized human lens epithelial (HLE) cells grown in vitro. METHODS: HLE cells, established from 18-week prenatal lenses, were maintained on bovine corneal endothelial (BCE) extracellular matrix (ECM) in medium supplemented with basic fibroblast growth factor (FGF-2). The identity, growth, and differentiation of the cultures were characterized by karyotyping, cell morphology, and growth kinetics studies, reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescence, and Western blot analysis. RESULTS: HLE cells had a male, human diploid (2N = 46) karyotype. The population-doubling time of exponentially growing cells was 24 hours. After 15 days in culture, cell morphology changed, and lentoid formation was evident. Reverse transcription-polymerase chain reaction (RT-PCR) indicated expression of alphaA- and betaB2-crystallin, fibroblast growth factor receptor 1 (FGFR1), and major intrinsic protein (MIP26) in exponential growth. Western analyses of protein extracts show positive expression of three immunologically distinct classes of crystallin proteins (alphaA-, alphaB-, and betaB2-crystallin) with time in culture. By Western blot analysis, expression of p57(KIP2), a known marker of terminally differentiated fiber cells, was detectable in exponential cultures, and levels increased after confluence. MIP26 and gamma-crystallin protein expression was detected in confluent cultures, by using immunofluorescence, but not in exponentially growing cells. CONCLUSIONS: HLE cells can be maintained for up to 4 months on ECM derived from BCE cells in medium containing FGF-2. With time in culture, the cells demonstrate morphologic characteristics of, and express protein markers for, lens fiber cell differentiation. This in vitro model will be useful for investigations of radiation-induced cataractogenesis and other studies of lens toxicity.

Development of a walleye spleen stromal cell line sensitive to viral hemorrhagic septicemia virus (VHSV IVb) and to protection by synthetic dsRNA.

PubMed

Vo, Nguyen T K; Bender, Aaron W; Ammendolia, Dustin A; Lumsden, John S; Dixon, Brian; Bols, Niels C

2015-07-01

A cell line, WE-spleen6, has been developed from the stromal layer of primary spleen cell cultures. On conventional plastic, WE-spleen6 cells had a spindle-shaped morphology at low cell density but grew to become epithelial-like at confluency. On the commercial extracellular matrix (ECM), Matrigel, the cells remained spindle-shaped and formed lumen-like structures. WE-spleen6 cells had intermediate filament protein, vimentin and the ECM protein, collagen I, but not smooth muscle α-actin (SMA) and von Willebrand factor (vWF) and lacked alkaline phosphatase and phagocytic activities. WE-spleen6 was more susceptible to infection with VHSV IVb than a fibroblast and epithelial cell lines from the walleye caudal fin, WE-cfin11f and WE-cfin11e, respectively. Viral transcripts and proteins appeared earlier in WE-spleen6 cultures as did cytopathic effect (CPE) and significant virus production. The synthetic double-stranded RNA (dsRNA), polyinosinic: polycytidylic acid (pIC), induced the antiviral protein Mx in both cell lines. Treating WE-spleen6 cultures with pIC prior to infection with VHSV IVb inhibited the early accumulation of viral transcripts and proteins and delayed the appearance of CPE and significant viral production. Of particular note, pIC caused the disappearance of viral P protein 2 days post infection. WE-spleen6 should be useful for investigating the impact of VHSV IVb on hematopoietic organs and the actions of pIC on the rhabdovirus life cycle. Copyright © 2015 Elsevier Ltd. All rights reserved.

Stepwise Synthesis of Giant Unilamellar Vesicles on a Microfluidic Assembly Line

PubMed Central

2011-01-01

Among the molecular milieu of the cell, the membrane bilayer stands out as a complex and elusive synthetic target. We report a microfluidic assembly line that produces uniform cellular compartments from droplet, lipid, and oil/water interface starting materials. Droplets form in a lipid-containing oil flow and travel to a junction where the confluence of oil and extracellular aqueous media establishes a flow-patterned interface that is both stable and reproducible. A triangular post mediates phase transfer bilayer assembly by deflecting droplets from oil, through the interface, and into the extracellular aqueous phase to yield a continuous stream of unilamellar phospholipid vesicles with uniform and tunable size. The size of the droplet precursor dictates vesicle size, encapsulation of small-molecule cargo is highly efficient, and the single bilayer promotes functional insertion of a bacterial transmembrane pore. PMID:21309555

Collaboration for rare disease drug discovery research.

PubMed

Litterman, Nadia K; Rhee, Michele; Swinney, David C; Ekins, Sean

2014-01-01

Rare disease research has reached a tipping point, with the confluence of scientific and technologic developments that if appropriately harnessed, could lead to key breakthroughs and treatments for this set of devastating disorders. Industry-wide trends have revealed that the traditional drug discovery research and development (R&D) model is no longer viable, and drug companies are evolving their approach. Rather than only pursue blockbuster therapeutics for heterogeneous, common diseases, drug companies have increasingly begun to shift their focus to rare diseases. In academia, advances in genetics analyses and disease mechanisms have allowed scientific understanding to mature, but the lack of funding and translational capability severely limits the rare disease research that leads to clinical trials. Simultaneously, there is a movement towards increased research collaboration, more data sharing, and heightened engagement and active involvement by patients, advocates, and foundations. The growth in networks and social networking tools presents an opportunity to help reach other patients but also find researchers and build collaborations. The growth of collaborative software that can enable researchers to share their data could also enable rare disease patients and foundations to manage their portfolio of funded projects for developing new therapeutics and suggest drug repurposing opportunities. Still there are many thousands of diseases without treatments and with only fragmented research efforts. We will describe some recent progress in several rare diseases used as examples and propose how collaborations could be facilitated. We propose that the development of a center of excellence that integrates and shares informatics resources for rare diseases sponsored by all of the stakeholders would help foster these initiatives.

Collaboration for rare disease drug discovery research

PubMed Central

Litterman, Nadia K.; Rhee, Michele; Swinney, David C.; Ekins, Sean

2014-01-01

Rare disease research has reached a tipping point, with the confluence of scientific and technologic developments that if appropriately harnessed, could lead to key breakthroughs and treatments for this set of devastating disorders. Industry-wide trends have revealed that the traditional drug discovery research and development (R&D) model is no longer viable, and drug companies are evolving their approach. Rather than only pursue blockbuster therapeutics for heterogeneous, common diseases, drug companies have increasingly begun to shift their focus to rare diseases. In academia, advances in genetics analyses and disease mechanisms have allowed scientific understanding to mature, but the lack of funding and translational capability severely limits the rare disease research that leads to clinical trials. Simultaneously, there is a movement towards increased research collaboration, more data sharing, and heightened engagement and active involvement by patients, advocates, and foundations. The growth in networks and social networking tools presents an opportunity to help reach other patients but also find researchers and build collaborations. The growth of collaborative software that can enable researchers to share their data could also enable rare disease patients and foundations to manage their portfolio of funded projects for developing new therapeutics and suggest drug repurposing opportunities. Still there are many thousands of diseases without treatments and with only fragmented research efforts. We will describe some recent progress in several rare diseases used as examples and propose how collaborations could be facilitated. We propose that the development of a center of excellence that integrates and shares informatics resources for rare diseases sponsored by all of the stakeholders would help foster these initiatives. PMID:25685324

Streamflow and water-quality conditions including geologic sources and processes affecting selenium loading in the Toll Gate Creek watershed, Aurora, Arapahoe County, Colorado, 2007

USGS Publications Warehouse

Paschke, Suzanne S.; Runkel, Robert L.; Walton-Day, Katherine; Kimball, Briant A.; Schaffrath, Keelin R.

2013-01-01

Toll Gate Creek is a perennial stream draining a suburban area in Aurora, Colorado, where selenium concentrations have consistently exceeded the State of Colorado aquatic-life standard for selenium of 4.6 micrograms per liter since the early 2000s. In cooperation with the City of Aurora, Colorado, Utilities Department, a synoptic water-quality study was performed along an 18-kilometer reach of Toll Gate Creek extending from downstream from Quincy Reservoir to the confluence with Sand Creek to develop a detailed understanding of streamflow and concentrations and loads of selenium in Toll Gate Creek. Streamflow and surface-water quality were characterized for summer low-flow conditions (July–August 2007) using four spatially overlapping synoptic-sampling subreaches. Mass-balance methods were applied to the synoptic-sampling and tracer-injection results to estimate streamflow and develop spatial profiles of concentration and load for selenium and other chemical constituents in Toll Gate Creek surface water. Concurrent groundwater sampling determined concentrations of selenium and other chemical constituents in groundwater in areas surrounding the Toll Gate Creek study reaches. Multivariate principal-component analysis was used to group samples and to suggest common sources for dissolved selenium and major ions. Hydrogen and oxygen stable-isotope ratios, groundwater-age interpretations, and chemical analysis of water-soluble paste extractions from core samples are presented, and interpretation of the hydrologic and geochemical data support conclusions regarding geologic sources of selenium and the processes affecting selenium loading in the Toll Gate Creek watershed.

Groundwater/surface-water interactions in the Tunk, Bonaparte, Antoine, and Tonasket Creek Subbasins, Okanogan River Basin, North-Central Washington, 2008

USGS Publications Warehouse

Sumioka, S.S.; Dinicola, R.S.

2009-01-01

An investigation into groundwater/surface-water interactions in four tributary subbasins of the Okanogan River determined that streamflows and shallow groundwater levels beneath the streams varied seasonally and by location. Streamflows measured in June 2008 indicated net losses of streamflow along 10 of 17 reaches, and hydraulic gradients measured between streams and shallow groundwater indicated potential recharge of surface water to groundwater at 11 of 21 measurement sites. In September 2008, net losses of streamflow were indicated along 9 of 17 reaches, and potential recharge of surface water to groundwater was indicated at 18 of 21 measurement sites. The greatest losses of streamflow occurred near the confluences with the Okanogan River, likely due to the presence of thick layers of unconsolidated deposits in the flood plain of the Okanogan River. Based on available geologic information compiled from drillers' logs, a surficial geologic map, and streamflow records, the extensive and thick deposits of unconsolidated material in the Tunk and Bonaparte Creek subbasins are factors in sustaining the almost perennial streamflow in those creeks. The less extensive and generally thinner unconsolidated deposits in the Tonasket and Antoine subbasins are contributing factors to the occasional extended periods of zero flow (a dry stream channel) in those creeks. Even though groundwater withdrawals would affect streamflows, relatively low precipitation in the area, along with limited groundwater storage capacity and the presence of permeable, unconsolidated deposits underlying the stream channels, would likely lead to loss of surface water to the groundwater system without any withdrawals.

Impacts of the Columbia River hydroelectric system on main-stem habitats of fall chinook salmon

USGS Publications Warehouse

Dauble, D.D.; Hanrahan, T.P.; Geist, D.R.; Parsley, M.J.

2003-01-01

Salmonid habitats in main-stem reaches of the Columbia and Snake rivers have changed dramatically during the past 60 years because of hydroelectric development and operation. Only about 13% and 58% of riverine habitats in the Columbia and Snake rivers, respectively, remain. Most riverine habitat is found in the upper Snake River; however, it is upstream of Hells Canyon Dam and not accessible to anadromous salmonids. We determined that approximately 661 and 805 km of the Columbia and Snake rivers, respectively, were once used by fall chinook salmon Oncorhynchus tshawytscha for spawning. Fall chinook salmon currently use only about 85 km of the main-stem Columbia River and 163 km of the main-stem Snake River for spawning. We used a geomorphic model to identify three river reaches downstream of present migration barriers with high potential for restoration of riverine processes: the Columbia River upstream of John Day Dam, the Columbia-Snake-Yakima River confluence, and the lower Snake River upstream of Little Goose Dam. Our analysis substantiated the assertion that historic spawning areas for fall chinook salmon occurred primarily within wide alluvial floodplains, which were once common in the mainstem Columbia and Snake rivers. These areas possessed more unconsolidated sediment and more bars and islands and had lower water surface slopes than did less extensively used areas. Because flows in the main stem are now highly regulated, the predevelopment alluvial river ecosystem is not expected to be restored simply by operational modification of one or more dams. Establishing more normative flow regimes - specifically, sustained peak flows for scouring - is essential to restoring the functional characteristics of existing, altered habitats. Restoring production of fall chinook salmon to any of these reaches also requires that population genetics and viability of potential seed populations (i.e., from tributaries, tailrace spawning areas, and hatcheries) be considered.

Simulation of wastewater effects on dissolved oxygen during low streamflow in the Red River of the North at Fargo, North Dakota, and Moorhead, Minnesota

USGS Publications Warehouse

Wesolowski, Edwin A.

1996-01-01

Pursuant to Section 303(d) of the Clean Water Act, both North Dakota and Minnesota identified part of the Red River of the North (Red River) as water-quality limited. The states are required to determine the total maximum daily load (TMDL) that can be discharged to a water-quality limited reach from various pollution sources without contravening water-quality standards (U.S. Environmental Protection Agency, 1991). A work group consisting of local, State, and Federal agency representatives that was organized in June 1994 decided that a TMDL should be developed in phases for a subreach of the Red River at Fargo, N. Dak., and Moorhead, Minn. (fig. 1). In the first phase, which is the basis for this report, the focus is on attainment of the instream dissolved-oxygen (DO) standard during low streamflows, and only Fargo and Moorhead wastewater-treatment-plant discharges and Sheyenne River inflow are considered. The study reach begins about 0.1 mile (mi) downstream (north) of the 12th Avenue North bridge in Fargo and extends 30.8 mi downstream to a site 0.8 mi upstream of the confluence of the Buffalo and Red Rivers (fig. 1). Nitrification of total ammonia (ammonia) from Fargo and Moorhead wastewater consumes most of the DO in the study reach (Wesolowski, 1994). Because the new (1995) Fargo plant already is nitrifying its wastewater, the work group needed to determine the maximum ammonia concentration for wastewater from the nonnitrifying Moorhead plant. To accomplish this task, the Red River at Fargo Water-Quality (RRatFGO QW) model (Wesolowski, 1994, 1996b) was used to simulate the effects of various wastewater-management alternatives during low streamflow. This report presents the results of those simulations to determine the usefulness of the model for management decisions. The simulations and report were completed in cooperation with the North Dakota Department of Health.

Lens epithelial cells derived from alphaB-crystallin knockout mice demonstrate hyperproliferation and genomic instability.

PubMed

Andley, U P; Song, Z; Wawrousek, E F; Brady, J P; Bassnett, S; Fleming, T P

2001-01-01

alphaB-crystallin is a member of the small heat shock protein family and can act as a molecular chaperone preventing the in vitro aggregation of other proteins denatured by heat or other stress conditions. Expression of alphaB-crystallin increases in cells exposed to stress and enhanced in tumors of neuroectodermal origin and in many neurodegenerative diseases. In the present study, we examined the properties of lens epithelial cells derived from mice in which the alphaB-crystallin gene had been knocked out. Primary rodent cells immortalize spontaneously in tissue culture with a frequency of 10(-5) to 10(-6). Primary lens epithelial cells derived from alphaB-crystallin-/- mice produced hyperproliferative clones at a frequency of 7.6 x 10(-2), four orders of magnitude greater than predicted by spontaneous immortalization (1). Hyperproliferative alphaB-crystallin-/- cells were shown to be truly immortal since they have been passaged for more than 100 population doublings without any diminution in growth potential. In striking contrast to the wild-type cells, which were diploid, the alphaB-crystallin-/- cultures had a high proportion of tetraploid and higher ploidy cells, indicating that the loss of alphaB-crystallin is associated with an increase in genomic instability. Further evidence of genomic instability of alphaB-crystallin-/- cells was observed when primary cultures were infected with Ad12-SV40 hybrid virus. In striking contrast to wild-type cells, alphaB-crystallin-/- cells expressing SV40 T antigen exhibited a widespread cytocidal response 2 to 3 days after attaining confluence, indicating that SV40 T antigen enhanced the intrinsic genomic instability of alphaB-crystallin-/- lens epithelial cells. These observations suggest that the widely distributed molecular chaperone alphaB-crystallin may play an important nuclear role in maintaining genomic integrity.

Theobroma cacao increases cells viability and reduces IL-6 and sVCAM-1 level in endothelial cells induced by plasma from preeclamptic patients.

PubMed

Rahayu, Budi; Baktiyani, Siti Candra Windu; Nurdiana, Nurdiana

2016-01-01

This study aims to investigate whether an ethanolic extract of Theobroma cacao bean is able to increase cell viability and decrease IL-6 and sVCAM-1 in endothelial cells induced by plasma from preeclamptic patients. Endothelial cells were obtained from human umbilical vascular endothelial cells. At confluency, endothelial cells were divided into six groups, which included control (untreated), endothelial cells exposed to plasma from normal pregnancy, endothelial cells exposed to 2% plasma from preeclamptic patients (PP), endothelial cells exposed to PP in the presence of ethanolic extract of T. cacao (PP+TC) at the following three doses: 25, 50, and 100 ppm. The analysis was performed in silico using the Hex 8.0, LigPlus and LigandScout 3.1 software. Analysis on IL-6 and sVCAM-1 levels were done by enzyme linked immunosorbent assay (ELISA). We found that seven of them could bind to the protein NFκB (catechin, leucoanthocyanidin, niacin, phenylethylamine, theobromine, theophylline, and thiamin). This increase in IL-6 was significantly (P<0.05) attenuated by both the 50 and 100 ppm treatments of T. cacao extract. Plasma from PP significantly increased sVCAM-1 levels compared to untreated cells. This increase in sVCAM-1 was significantly attenuated by all doses of the extract. In conclusion, T. cacao extract prohibits the increase in IL-6 and sVCAM-1 in endothelial cells induced by plasma from preeclamptic patients. Therefore this may provide a herbal therapy for attenuating the endothelial dysfunction found in preeclampsia. Copyright © 2016 International Society for the Study of Hypertension in Pregnancy. Published by Elsevier B.V. All rights reserved.

Characteristics and EGFP expression of goat mammary gland epithelial cells.

PubMed

Zheng, Y-M; He, X-Y; Zhang, Y

2010-12-01

The aims of this study were (i) to establish a goat mammary gland epithelial (GMGE) cell line, and (ii) to determine if these GMGE cells could be maintained long-term in culture by continuous subculturing following transfection with a reporter gene, enhanced green fluorescence protein (EGFP). Primary culture of GMGE cells was achieved by outgrowth of migrating cells from the fragments of the mammary gland tissue of a lactating goat. The passage 16 GMGE cells were transfected with EGFP gene using lipofection. The expression of Cell keratins of epithelial cells in GMGE cells was test by immunofluorescence. Βeta-Casein gene mRNA was test for GMGE cells by RT-PCR. The results showed that when grown at low density on a plastic substratum, the GMGE cells formed islands, and when grown to confluency, the cells formed a monolayer and aggregated with the characteristic cobble-stone morphology of epithelial cells. GMGE cells could form dome-like structure which looked like nipple, and the lumen-like structures formed among the cells. Several blister-like structures appeared in the appearance of the cells. The GMGE cells contained different cell types, majority of the cells were short shuttle-like or polygon which were beehive-like. A part of cells were round and flat, a small number of cells were elongated. Some of the GMGE cells contained milk drops. The cell nuclei were round which had 2-4 obvious cores. The expression of Cell keratins demonstrated the property of epithelial cells in GMGE cells by immunofluorescence. The GMGE cells could express transcript encoding a Βeta-Casein protein. EGFP gene was successfully transferred into the GMGE cells, and the transfected cells could be maintained long-term in culture by continuous subculturing. In conclusion, we have established a EGFP gene transfected GMGE (ET-GMGE) cell line and maintained it long-term in culture by continuous subculturing. © 2010 Blackwell Verlag GmbH.

75 FR 68738 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-11-09

... +156 +155 upstream of Taylors Store Road (Secondary Road 1004). Polecat Branch At the confluence with...). Just upstream of U.S. +151 +152 Route 64. Swift Creek Approximately 1.8 miles +90 +88 City of Rocky...

Unit 6, downstream from Ferndale Bridge Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 6, downstream from Ferndale Bridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 5, upstream toward incline bridge Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 5, upstream toward incline bridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 2, downstream from Coppersdale Bridge Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 2, downstream from Coppersdale Bridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Elevated view of city from incline Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Elevated view of city from incline - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 3, downstream from Point Park Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 3, downstream from Point Park - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 1, downstream from Laurel Run Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 1, downstream from Laurel Run - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 3, upstream from footbridge Johnstown Local Flood Protection ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 3, upstream from footbridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

4. NORTH ELEVATION OF POWER PLANT LOOKING SOUTH SOUTHWEST. ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

4. NORTH ELEVATION OF POWER PLANT LOOKING SOUTH SOUTHWEST. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

11. EAST WALL OF POWER PLANT BUILDING LOOKING WEST. ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

11. EAST WALL OF POWER PLANT BUILDING LOOKING WEST. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

40 CFR 140.4 - Complete prohibition.

Code of Federal Regulations, 2011 CFR

2011-07-01

... Florida within the boundaries of the Florida Keys National Marine Sanctuary as delineated on a map of the... State, the first is bounded by an east-west line through the most northern confluence of the Mohawk...

76 FR 21857 - Endangered and Threatened Species; Take of Anadromous Fish

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-19

... Klamath River and estuary, and at the confluence of Independence Creek and the Klamath River. USGS... the estuary and will consist of capture (beach seine), handle (identify, measure, weigh), and release...

78 FR 20942 - Niobrara Confluence and Ponca Bluffs Conservation Areas, NE and SD; Draft Environmental Impact...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-04-08

... that remains unchannelized, relatively free-flowing, and undeveloped. This area of the Missouri River's... resources to enhance conservation; enhance recreation; increase tourism; instill new money into local...

10. Detail of Derrick Foot Showing Pipe Construction, Looking Southwest ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

10. Detail of Derrick Foot Showing Pipe Construction, Looking Southwest - David Renfrew Oil Rig, East side of Connoquenessing Creek, 0.4 mile North of confluence with Thorn Creek, Renfrew, Butler County, PA

76 FR 50960 - Proposed Flood Elevation Determinations

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-17

... 1.5 miles None +65 upstream of Beaman Old Creek Road. Cow Branch At the Nahunta Swamp +61 +60 Unincorpo rated Areas confluence. of Greene County. Approximately 2.1 miles None +114 upstream of Cow Branch...

The emergence of the nanobiotechnology industry

NASA Astrophysics Data System (ADS)

Maine, Elicia; Thomas, V. J.; Bliemel, Martin; Murira, Armstrong; Utterback, James

2014-01-01

The confluence of nanotechnology and biotechnology provides significant commercial opportunities. By identifying, classifying and tracking firms with capabilities in both biotechnology and nanotechnology over time, we analyse the emergence and evolution of the global nanobiotechnology industry.

Electrical Detection of Cancer Biomarker using Aptamers with Nanogap Break-Junctions

PubMed Central

Ilyas, Azhar; Asghar, Waseem; Allen, Peter B.; Duhon, Holli; Ellington, Andrew D.; Iqbal, Samir M.

2012-01-01

Epidermal Growth Factor Receptor (EGFR) is a cell surface protein overexpressed in cancerous cells. It is known to be the most common oncongene. EGFR concentration also increases in the serum of cancer patients. The detection of small changes in the concentration of EGFR can be critical for early diagnosis, resulting in better treatment and improved survival rate of cancer patients. This article reports an RNA aptamer based approach to selectively capture EGFR protein and an electrical scheme for its detection. Pairs of gold electrodes with nanometer separation were made through confluence of focused ion beam scratching and electromigration. The aptamer was hybridized to a single stranded DNA molecule, which in turn was immobilized on SiO2 surface between the gold nanoelectrodes. The selectivity of the aptamer was demonstrated by using control chips with mutated non–selective aptamer and with no aptamer. Surface functionalization was characterized by optical detection and two orders of magnitude increase in direct current (DC) was measured when selective capture of EGFR occurred. This represents an electronic biosensor for the detection of proteins of interest for medical applications. PMID:22706642

Spectral phasor analysis of LAURDAN fluorescence in live A549 lung cells to study the hydration and time evolution of intracellular lamellar body-like structures.

PubMed

Malacrida, Leonel; Astrada, Soledad; Briva, Arturo; Bollati-Fogolín, Mariela; Gratton, Enrico; Bagatolli, Luis A

2016-11-01

Using LAURDAN spectral imaging and spectral phasor analysis we concurrently studied the growth and hydration state of subcellular organelles (lamellar body-like, LB-like) from live A549 lung cancer cells at different post-confluence days. Our results reveal a time dependent two-step process governing the size and hydration of these intracellular LB-like structures. Specifically, a first step (days 1 to 7) is characterized by an increase in their size, followed by a second one (days 7 to 14) where the organelles display a decrease in their global hydration properties. Interestingly, our results also show that their hydration properties significantly differ from those observed in well-characterized artificial lamellar model membranes, challenging the notion that a pure lamellar membrane organization is present in these organelles at intracellular conditions. Finally, these LB-like structures show a significant increase in their hydration state upon secretion, suggesting a relevant role of entropy during this process. Copyright © 2016 Elsevier B.V. All rights reserved.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Varley, Claire; Hill, Gemma; Pellegrin, Stephanie

Regeneration of the urothelium is rapid and effective in order to maintain a barrier to urine following tissue injury. Whereas normal human urothelial (NHU) cells are mitotically quiescent and G0 arrested in situ, they rapidly enter the cell cycle upon seeding in primary culture and show reversible growth arrest at confluency. We have used this as a model to investigate the role of EGF receptor signaling in urothelial regeneration and wound-healing. Transcripts for HER-1, HER-2, and HER-3 were expressed by quiescent human urothelium in situ. Expression of HER-1 was upregulated in proliferating cultures, whereas HER-2 and HER-3 were more associatedmore » with a growth-arrested phenotype. NHU cells could be propagated in the absence of exogenous EGF, but autocrine signaling through HER-1 via the MAPK and PI3-kinase pathways was essential for proliferation and migration during urothelial wound repair. HB-EGF was expressed by urothelium in situ and HB-EGF, epiregulin, TGF-{alpha}, and amphiregulin were expressed by proliferating NHU cells. Urothelial wound repair in vitro was attenuated by neutralizing antibodies against HER-1 ligands, particularly amphiregulin. By contrast, the same ligands applied exogenously promoted migration, but inhibited proliferation, implying that HER-1 ligands provoke differential effects in NHU cells depending upon whether they are presented as soluble or juxtacrine ligands. We conclude that proliferation and migration during wound healing in NHU cells are mediated through an EGFR autocrine signalling loop and our results implicate amphiregulin as a key mediator.« less

Galectin-8 induces partial epithelial–mesenchymal transition with invasive tumorigenic capabilities involving a FAK/EGFR/proteasome pathway in Madin–Darby canine kidney cells

PubMed Central

Oyanadel, Claudia; Holmes, Christopher; Pardo, Evelyn; Retamal, Claudio; Shaughnessy, Ronan; Smith, Patricio; Cortés, Priscilla; Bravo-Zehnder, Marcela; Metz, Claudia; Feuerhake, Teo; Romero, Diego; Roa, Juan Carlos; Montecinos, Viviana; Soza, Andrea; González, Alfonso

2018-01-01

Epithelial cells can acquire invasive and tumorigenic capabilities through epithelial–mesenchymal-transition (EMT). The glycan-binding protein galectin-8 (Gal-8) activates selective β1-integrins involved in EMT and is overexpressed by certain carcinomas. Here we show that Gal-8 overexpression or exogenous addition promotes proliferation, migration, and invasion in nontumoral Madin–Darby canine kidney (MDCK) cells, involving focal-adhesion kinase (FAK)-mediated transactivation of the epidermal growth factor receptor (EGFR), likely triggered by α5β1integrin binding. Under subconfluent conditions, Gal-8–overexpressing MDCK cells (MDCK-Gal-8H) display hallmarks of EMT, including decreased E-cadherin and up-regulated expression of vimentin, fibronectin, and Snail, as well as increased β-catenin activity. Changes related to migration/invasion included higher expression of α5β1 integrin, extracellular matrix-degrading MMP13 and urokinase plasminogen activator/urokinase plasminogen activator receptor (uPA/uPAR) protease systems. Gal-8–stimulated FAK/EGFR pathway leads to proteasome overactivity characteristic of cancer cells. Yet MDCK-Gal-8H cells still develop apical/basolateral polarity reverting EMT markers and proteasome activity under confluence. This is due to the opposite segregation of Gal-8 secretion (apical) and β1-integrins distribution (basolateral). Strikingly, MDCK-Gal-8H cells acquired tumorigenic potential, as reflected in anchorage-independent growth in soft agar and tumor generation in immunodeficient NSG mice. Therefore, Gal-8 can promote oncogenic-like transformation of epithelial cells through partial and reversible EMT, accompanied by higher proliferation, migration/invasion, and tumorigenic properties. PMID:29298841

SASH1 mediates sensitivity of breast cancer cells to chloropyramine and is associated with prognosis in breast cancer

PubMed Central

Burgess, Joshua T.; Bolderson, Emma; Saunus, Jodi M.; Zhang, Shu-Dong; Reid, Lynne E.; McNicol, Anne Marie; Lakhani, Sunil R.; Cuff, Katharine; Richard, Kerry; Richard, Derek J.; O'Byrne, Kenneth J.

2016-01-01

Expression of the SASH1 protein is reduced in a range of human cancers and has been implicated in apoptotic cancer cell death. This study investigated whether increasing SASH1 expression could be a useful therapeutic strategy in breast cancer. Ectopic SASH1 expression increased apoptosis in 7/8 breast cancer cell lines. Subsequent in silico connectivity screening demonstrated that the clinically approved antihistamine drug, chloropyramine, increased SASH1 mRNA levels. Chloropyramine has previously been shown to have anti-tumour activity in breast cancer in part through modulation of FAK signalling, a pathway also regulated by SASH1. This study demonstrated that chloropyramine increased SASH1 protein levels in breast cancer cells. Consistent with this the agent reduced cell confluency in 7/8 cell lines treated irrespective of their ER status but not apoptosis incompetent MCF7 cells. In contrast SASH1 siRNA-transfected breast cancer cells exhibited reduced chloropyramine sensitivity. The prognostic significance of SASH1 expression was also investigated in two breast cancer cohorts. Expression was associated with favourable outcome in ER-positive cases, but only those of low histological grade/proliferative status. Conversely, we found a very strong inverse association in HER2+ disease irrespective of ER status, and in triple-negative, basal-like cases. Overall, the data suggest that SASH1 is prognostic in breast cancer and could have subtype-dependent effects on breast cancer progression. Pharmacologic induction of SASH1 by chloropyramine treatment of breast cancer warrants further preclinical and clinical investigation. PMID:27637080

SASH1 mediates sensitivity of breast cancer cells to chloropyramine and is associated with prognosis in breast cancer.

PubMed

Burgess, Joshua T; Bolderson, Emma; Saunus, Jodi M; Zhang, Shu-Dong; Reid, Lynne E; McNicol, Anne Marie; Lakhani, Sunil R; Cuff, Katharine; Richard, Kerry; Richard, Derek J; O'Byrne, Kenneth J

2016-11-08

Expression of the SASH1 protein is reduced in a range of human cancers and has been implicated in apoptotic cancer cell death. This study investigated whether increasing SASH1 expression could be a useful therapeutic strategy in breast cancer. Ectopic SASH1 expression increased apoptosis in 7/8 breast cancer cell lines. Subsequent in silico connectivity screening demonstrated that the clinically approved antihistamine drug, chloropyramine, increased SASH1 mRNA levels. Chloropyramine has previously been shown to have anti-tumour activity in breast cancer in part through modulation of FAK signalling, a pathway also regulated by SASH1. This study demonstrated that chloropyramine increased SASH1 protein levels in breast cancer cells. Consistent with this the agent reduced cell confluency in 7/8 cell lines treated irrespective of their ER status but not apoptosis incompetent MCF7 cells. In contrast SASH1 siRNA-transfected breast cancer cells exhibited reduced chloropyramine sensitivity. The prognostic significance of SASH1 expression was also investigated in two breast cancer cohorts. Expression was associated with favourable outcome in ER-positive cases, but only those of low histological grade/proliferative status. Conversely, we found a very strong inverse association in HER2+ disease irrespective of ER status, and in triple-negative, basal-like cases. Overall, the data suggest that SASH1 is prognostic in breast cancer and could have subtype-dependent effects on breast cancer progression. Pharmacologic induction of SASH1 by chloropyramine treatment of breast cancer warrants further preclinical and clinical investigation.

Type I collagen-induced YAP nuclear expression promotes primary cilia growth and contributes to cell migration in confluent mouse embryo fibroblast 3T3-L1 cells.

PubMed

Xu, Qian; Liu, Xiaoling; Liu, Weiwei; Hayashi, Toshihiko; Yamato, Masayuki; Fujisaki, Hitomi; Hattori, Shunji; Tashiro, Shin-Ichi; Onodera, Satoshi; Ikejima, Takashi

2018-05-30

The extracellular matrix (ECM) is a major biomechanical environment for all cells in vivo, and tightly controls wound healing and cancer progression. Type I collagen (Col I) is the most abundant component in ECM and plays an essential role for cell motility control and migration beyond structural support. Our previous results showed that Col I increased the length of primary cilia and the expression of primary cilia-associated proteins in 3T3-L1 cells. The Hippo/YAP pathway serves as a major integrator of cell surface-mediated signals and regulates key processes for the development and maintenance of tissue functions. In this study, we investigated the role of Hippo/YAP signaling in primary cilia growth of cells cultured on Col I-coated plate, as well as the potential link between primary cilia and migration. At 2-day post-confluence, YAP localization in the nucleus was dramatically increased when the cells were cultured on Col I-coated plate, accompanied by cilia growth. YAP inhibitor verteporfin repressed the growth of primary cilia as well as the expressions of ciliogenesis-associated proteins in confluent 3T3-L1 cells cultured on Col I-coated plate. Moreover, knockdown of either YAP or IFT88, one of the ciliogenesis-associated proteins, reversed the migration of confluent 3T3-L1 cells promoted by Col I-coating. In conclusion, activation of YAP pathway by Col I-coating of culture plate for confluent 3T3-L1 cells is positively associated with the primary cilia growth, which eventually results in promoted migration.

Thermal and overcharge abuse analysis of a redox shuttle for overcharge protection of LiFePO4

NASA Astrophysics Data System (ADS)

Lamb, Joshua; Orendorff, Christopher J.; Amine, Khalil; Krumdick, Gregory; Zhang, Zhengcheng; Zhang, Lu; Gozdz, Antoni S.

2014-02-01

This work investigated the performance and abuse tolerance of cells protected using the redox shuttle 1,4-bis(2-methoxyethoxy)-2,5-di-tert-butylbenzene. The thermal efficiencies were evaluated using isothermal battery calorimetry. Cells containing the overcharge shuttle were observed to reach a steady state value of approximately 3.8 V, with a small variance in direct proportion to the applied current. In all cases the heat output from the cells was measured to reach ∼90% of the total input power. The heat output was also measured using isothermal calorimetry. At higher rates of overcharge, the data shows that the cell containing the shuttle rapidly reaches a steady state voltage, while the temperature increases until a moderately high steady state temperature is reached. The control cell meanwhile rapidly increases in both applied voltage and cell temperature until cell failure. Two cells in series were taken deliberately out of balance individually, then charged as a single pack to observe the time needed to bring the cells into balance with one another.

Travel Times, Streamflow Velocities, and Dispersion Rates in the Yellowstone River, Montana

USGS Publications Warehouse

McCarthy, Peter M.

2009-01-01

The Yellowstone River is a vital natural resource to the residents of southeastern Montana and is a primary source of water for irrigation and recreation and the primary source of municipal water for several cities. The Yellowstone River valley is the primary east-west transportation corridor through southern Montana. This complex of infrastructure makes the Yellowstone River especially vulnerable to accidental spills from various sources such as tanker cars and trucks. In 2008, the U.S. Geological Survey (USGS), in cooperation with the Montana Department of Environmental Quality, initiated a dye-tracer study to determine instream travel times, streamflow velocities, and dispersion rates for the Yellowstone River from Lockwood to Glendive, Montana. The purpose of this report is to describe the results of this study and summarize data collected at each of the measurement sites between Lockwood and Glendive. This report also compares the results of this study to estimated travel times from a transport model developed by the USGS for a previous study. For this study, Rhodamine WT dye was injected at four locations in late September and early October 2008 during reasonably steady streamflow conditions. Streamflows ranged from 3,490 to 3,770 cubic feet per second upstream from the confluence of the Bighorn River and ranged from 6,520 to 7,570 cubic feet per second downstream from the confluence of the Bighorn River. Mean velocities were calculated for each subreach between measurement sites for the leading edge, peak concentration, centroid, and trailing edge at 10 percent of the peak concentration. Calculated velocities for the centroid of the dye plume for subreaches that were completely laterally mixed ranged from 1.83 to 3.18 ft/s within the study reach from Lockwood Bridge to Glendive Bridge. The mean of the completely mixed centroid velocity for the entire study reach, excluding the subreach between Forsyth Bridge and Cartersville Dam, was 2.80 ft/s. Longitudinal dispersion rates of the dye plume for this study ranged from 0.06 ft/s for the subreach upstream from Forsyth Bridge to 2.25 ft/s for the subreach upstream from Calyspo Bridge for subreaches where the dye was completely laterally mixed. A relation was determined between travel time of the peak concentration and time for the dye plume to pass a site (duration). This relation can be used to estimate when the receding concentration of a potential contaminant reaches 10 percent of its peak concentration for accidental spills into the Yellowstone River. Data from this dye-tracer study were used to evaluate velocity and concentration estimates from a transport model developed as part of an earlier USGS study. Comparison of the estimated and calculated velocities for the study reach indicate that the transport model estimates the velocities of the Yellowstone River between Huntley Bridge and Glendive Bridge with reasonable accuracy. Velocities of the peak concentration of the dye plume calculated for this study averaged 10 percent faster than the most probable velocities and averaged 12 percent slower than the maximum probable velocities estimated from the transport model. Peak Rhodamine WT dye concentrations were consistently lower than the transport model estimates except for the most upstream subreach of each dye injection. The most upstream subreach of each dye injection is expected to have a higher concentration because of incomplete lateral mixing. Lower measured peak concentrations for all other sites were expected because Rhodamine WT dye deteriorates when exposed to sunlight and will sorb onto the streambanks and stream bottom. Velocity-streamflow relations developed by using routine streamflow measurements at USGS gaging stations and the transport model can be used to estimate mean streamflow velocities throughout a range of streamflows. The variation in these velocity-streamflow relations emphasizes the uncertainty in estimating the mean streamflow veloc

The influence of network structure upon sediment routing in two disturbed catchments, East Cape, New Zealand

NASA Astrophysics Data System (ADS)

Walley, Yasmin; Tunnicliffe, Jon; Brierley, Gary

2018-04-01

Lateral inputs from hillslopes and tributaries exert a variable impact upon the longitudinal connectivity of sediment transfer in river systems with differing drainage network configurations. Network topology influences channel slope and confinement at confluence zones, thereby affecting patterns of sediment storage and the conveyance of sediments through catchments. Rates of disturbance response, patterns of sediment propagation, and the implications for connectivity and recovery were assessed in two neighbouring catchments with differing network configurations on the East Cape of New Zealand. Both catchments were subject to forest clearing in the late 1940s and a major cyclonic storm in 1988. However, reconstruction of landslide runout pathways, and characterization of connectivity using a Tokunaga framework, demonstrates different patterns and rates of sediment transfer and storage in a dendritic network relative to a more elongate, herringbone drainage network. The dendritic network has a higher rate of sediment transfer between storage sites in successive Strahler orders, whereas longitudinal connectivity along the fourth-order mainstem is disrupted by lateral sediment inputs from multiple low-order tributaries in the more elongate, herringbone network. In both cases the most dynamic ('hotspot') reaches are associated with a high degree of network side-branching.

Flooding in the South Platte River and Fountain Creek Basins in eastern Colorado, September 9–18, 2013

USGS Publications Warehouse

Kimbrough, Robert A.; Holmes, Robert R.

2015-11-25

Flooding in the Fountain Creek Basin was primarily contained to Fountain Creek from southern Colorado Springs to its confluence with the Arkansas River in Pueblo, in lower Monument Creek, and in several mountain tributaries. New record peak streamflows occurred at four mountain tributary streamgages having at least 10 years of record; Bear Creek, Cheyenne Creek, Rock Creek, and Little Fountain Creek. Five streamgages with at least 10 years of record in a 32-mile reach of Fountain Creek extending from Colorado Springs to Piñon had peak streamflows in the top five for the period of record. A peak of 15,300 ft3/s at Fountain Creek near Fountain was the highest streamflow recorded in the Fountain Creek Basin during the September 2013 event and ranks the third highest peak in 46 years. Near the mouth of the basin, a peak of 11,800 ft3/s in Pueblo was only the thirteenth highest annual peak in 74 years. A new Colorado record for daily rainfall of 11.85 inches was recorded at a USGS rain gage in the Little Fountain Creek Basin on September 12, 2013.

A novel land surface-hydrologic-sediment dynamics model for stream corridor conservation assessment and its first application

NASA Astrophysics Data System (ADS)

Smithgall, K.; Shen, C.; Langendoen, E. J.; Johnson, P. A.

2015-12-01

Nationally and in the Chesapeake Bay (CB), Stream Corridor restoration costs unsustainable amount of public resources, but decisions are often made with inadequate knowledge of regional-scale system behavior. Bank erosion is a significant issue relevant to sediment and nutrient pollution, aquatic and riparian habitat and stream health. Existing modeling effort either focuses only on reach-scale responses or overly simplifies the descriptions for bank failure mechanics. In this work we present a novel regional-scale processes model integrating hydrology, vegetation dynamics, hydraulics, bank mechanics and sediment transport, based on a coupling between Community Land Model, Process-based Adaptive Watershed Simulator and CONservational Channel Evolution and Pollutant Transport System (CLM + PAWS + CONCEPTS, CPC). We illustrate the feasibility of this modeling platform in a Valley and Ridge basin in Pennsylvania, USA, with channel geometry data collected in 2004 and 2014. The simulations are able to reproduce essential pattern of the observed trends. We study the causes of the noticeable evolution of a relocated channel and the hydrologic controls. Bridging processes on multiple scales, the CPC model creates a new, integrated system that may serve as a confluence point for inter-disciplinary research.

Catastrophic flood disturbance and a community's response to plant resilience in the heart of the Texas Hill Country

NASA Astrophysics Data System (ADS)

Meitzen, Kimberly M.; Phillips, John N.; Perkins, Thaïs; Manning, Aspen; Julian, Jason P.

2018-03-01

The Blanco River, which flows through the limestone Balcones Canyonlands of central Texas (USA), experienced catastrophic flooding in May 2015 that resulted in significant biogeomorphic disturbance to its riparian corridor. High-resolution aerial and satellite imagery from pre- and post-flooding for a 55-km reach of river were used to map and categorize patterns of disturbance by degree of severity ranging from complete floodplain stripping to no disturbance. The most severe disturbance occurred within the floodway near the channel and decreased with lateral distance into the 100- and 500-year floodplains. Disturbance patterns previously identified in the literature including meander scour, parallel chute scour, convex bank erosion, and macroturbulent scour were all present following this event, as well as substantial disturbance proximal to tributary confluences. In the aftermath of this event, TreeFolks, a local nonprofit organization, engaged with the community to actively replant and restore the riparian corridor of the Blanco River on public and private lands. These reforestation efforts supplement the natural passive recovery of the riparian corridor, enabling the system to recover more quickly and be resilient to future flood events.

Unit 5, downstream from Hickory Street Bridge Johnstown Local ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 5, downstream from Hickory Street Bridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 6, downstream from Horner Street Bridge Johnstown Local ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 6, downstream from Horner Street Bridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 3, downstream from Fourth Avenue Bridge Johnstown Local ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 3, downstream from Fourth Avenue Bridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 4, upstream from Johns Street Bridge Johnstown Local ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 4, upstream from Johns Street Bridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 5, downstream from Haynes Street Bridge Johnstown Local ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 5, downstream from Haynes Street Bridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 4, downstream from Johns Street Bridge Johnstown Local ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 4, downstream from Johns Street Bridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 4, downstream from First Street Bridge Johnstown Local ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 4, downstream from First Street Bridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 6, upstream from Hickory Street Bridge Johnstown Local ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 6, upstream from Hickory Street Bridge - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

10. WEST WALL OF POWER PLANT BUILDING LOOKING EAST SOUTHEAST. ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

10. WEST WALL OF POWER PLANT BUILDING LOOKING EAST SOUTHEAST. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

14. INTERIOR OF POWER PLANT LOOKING SOUTHEAST AT ELECTRICAL PANEL. ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

14. INTERIOR OF POWER PLANT LOOKING SOUTHEAST AT ELECTRICAL PANEL. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

12. CANAL SLUICE GATE LOCATED 150' WEST OF POWER PLANT. ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

12. CANAL SLUICE GATE LOCATED 150' WEST OF POWER PLANT. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

17. TOP OF TURBINE LOOKING NORTHWEST. SHAFT TO GENERATOR IN ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

17. TOP OF TURBINE LOOKING NORTHWEST. SHAFT TO GENERATOR IN CENTER. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

9. Mispillion Lighthouse, Tower Lantern Floor Hatch Mispillion Lighthouse, ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

9. Mispillion Lighthouse, Tower Lantern Floor Hatch - Mispillion Lighthouse, South bank of Mispillion River at its confluence with Delaware River at northeast end of County Road 203, 7 miles east of Milford, Milford, Sussex County, DE

Sacramento River Water Treatment Plant Intake Pier & Access Bridge, ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Sacramento River Water Treatment Plant Intake Pier & Access Bridge, Spanning Sacramento River approximately 175 feet west of eastern levee on river; roughly .5 mile downstream from confluence of Sacramento & American Rivers, Sacramento, Sacramento County, CA

13. Bull Wheel Core Showing Name of Manufacturer (National Supply ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

13. Bull Wheel Core Showing Name of Manufacturer (National Supply Co.), Looking Southwest - David Renfrew Oil Rig, East side of Connoquenessing Creek, 0.4 mile North of confluence with Thorn Creek, Renfrew, Butler County, PA

7. Band Wheel, Showing Cloth Web Belt and Wooden Bearing ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

7. Band Wheel, Showing Cloth Web Belt and Wooden Bearing Blocks, Looking West - David Renfrew Oil Rig, East side of Connoquenessing Creek, 0.4 mile North of confluence with Thorn Creek, Renfrew, Butler County, PA

9. Detail Showing top of Samson Post and Walking Beam ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

9. Detail Showing top of Samson Post and Walking Beam Pivot, Looking East - David Renfrew Oil Rig, East side of Connoquenessing Creek, 0.4 mile North of confluence with Thorn Creek, Renfrew, Butler County, PA

Artificial Intelligence and Vocational Education: An Impending Confluence.

ERIC Educational Resources Information Center

Roth, Gene L.; McEwing, Richard A.

1986-01-01

Reports on the relatively new field of artificial intelligence and its relationship to vocational education. Compares human intelligence with artificial intelligence. Discusses expert systems, natural language technology, and current trends. Lists potential applications for vocational education. (CH)

Phytoplankton bloom in Persian Gulf

NASA Technical Reports Server (NTRS)

2002-01-01

There is a large amount of sediment clearly visible in the true-color image of the Persian Gulf, acquired on November 1, 2001, by MODIS. Carried by the confluence of the Tigris and Euphrates Rivers (at center), the sediment-laden waters appear light brown where they enter the northern end of the Persian Gulf and then gradually dissipate into turquoise swirls as they drift southward. The nutrients these sediments carry are helping to support a phytoplankton bloom in the region, which adds some darker green hues in the rich kaleidoscope of colors on the surface (see the high resolution image). The confluence of the Tigris and Euphrates Rivers marks the southernmost boundary between Iran (upper right) and Iraq (upper left). South of Iraq are the countries of Kuwait and Saudi Arabia. The red dots indicate the probable locations of fires burning at oil refineries. Thin black plumes of smoke can be seen streaming away from several of these.

An Experimental Investigation of the Confluent Boundary Layer on a High-Lift System

NASA Technical Reports Server (NTRS)

Thomas, F. O.; Nelson, R. C.

1997-01-01

This paper describes a fundamental experimental investigation of the confluent boundary layer generated by the interaction of a leading-edge slat wake with the boundary layer on the main element of a multi-element airfoil model. The slat and airfoil model geometry are both fully two-dimensional. The research reported in this paper is performed in an attempt to investigate the flow physics of confluent boundary layers and to build an archival data base on the interaction of the slat wake and the main element wall layer. In addition, an attempt is made to clearly identify the role that slat wake / airfoil boundary layer confluence has on lift production and how this occurs. Although complete LDV flow surveys were performed for a variety of slat gap and overhang settings, in this report the focus is on two cases representing both strong and weak wake boundary layer confluence.

Trends in the Brazil/Malvinas Confluence region

NASA Astrophysics Data System (ADS)

Combes, Vincent; Matano, Ricardo P.

2014-12-01

Observations show abrupt changes in the oceanic circulation of the southwestern Atlantic. These studies report a southward drift of the Brazil/Malvinas Confluence (BMC) and a change in the spectral characteristics of the Malvinas Current (MC) transport. We address the cause of these changes using the result of a high-resolution numerical experiment. The experiment, which is consistent with observations, shows a southward BMC displacement at a rate of 0.62°/decade between 1993 and 2008, and a shift of the spectral characteristics of the MC transport after 1999. We find that these changes are driven by a weakening of the northern branch of the Antarctic Circumpolar Current, which translates to a weakening of the MC transport and a southward BMC drift. The drift changes the spectral characteristics of the MC transport, which becomes more influenced by annual and semiannual variations associated with the BMC.

Why Do Some Men Misperceive Women’s Sexual Intentions More Frequently Than Others Do? An Application of the Confluence Model

PubMed Central

Jacques-Tiura, Angela J.; Abbey, Antonia; Parkhill, Michele R.; Zawacki, Tina

2015-01-01

Although many researchers have documented men’s tendency to misperceive women’s friendliness as a sign of sexual interest, few have examined individual differences in men’s attitudes and past experiences that might predict their likelihood of making these types of misjudgments. We applied an expanded version of Malamuth, Sockloskie, Koss, and Tanaka’s (1991) Confluence model to predict frequency of misperception of women’s sexual intent with a sample of 356 male college students. Using structural equation modeling, hostile masculinity, impersonal sex, and drinking in dating and sexual situations predicted men’s frequency of misperception. Furthermore, the more risk factors men possessed, the more times they misperceived women’s sexual intentions. Suggestions are made for theory development and future research incorporating situational as well as personality measures in longitudinal studies. PMID:17933743

Apparatus And Method For Reducing Drag Of A Bluff Body In Ground Effect Using Counter-Rotating Vortex Pairs

DOEpatents

Ortega, Jason M.; Sabari, Kambiz

2005-12-27

An aerodynamic base drag reduction apparatus and method for bluff bodies, such as tractor-trailer trucks, utilizing a pair of lift surfaces extending to lift surface tips and located alongside the bluff body such as on opposing left and right side surfaces. In a flowstream substantially parallel to the longitudinal centerline of the bluff body, the pair of lift surfaces generate a pair of counter-rotating trailing vortices which confluence together in the wake of the bluff body in a direction orthogonal to the flowstream. The confluence draws or otherwise turns the flowstream, such as the flowstream passing over a top surface of the bluff body, in and around behind a trailing end of the bluff body to raise the pressure on a base surface at the trailing end and thereby reduce the aerodynamic base drag.

Apparatus And Method For Reducing Drag Of A Bluff Body In Ground Effect Using Counter-Rotating Vortex Pairs

DOEpatents

Ortega, Jason M.; Salari, Kambiz

2005-08-09

An aerodynamic base drag reduction apparatus and method for bluff bodies, such as tractor-trailer trucks, utilizing a pair of lift surfaces extending to lift surface tips and located alongside the bluff body such as on opposing left and right side surfaces. In a flowstream substantially parallel to the longitudinal centerline of the bluff body, the pair of lift surfaces generate a pair of counter-rotating trailing vortices which confluence together in the wake of the bluff body in a direction orthogonal to the flowstream. The confluence draws or otherwise turns the flowstream, such as the flowstream passing over a top surface of the bluff body, in and around behind a trailing end of the bluff body to raise the pressure on a base surface at the trailing end and thereby reduce the aerodynamic base drag.

Modeling Cancer Cell Growth Dynamics In vitro in Response to Antimitotic Drug Treatment

PubMed Central

Lorz, Alexander; Botesteanu, Dana-Adriana; Levy, Doron

2017-01-01

Investigating the role of intrinsic cell heterogeneity emerging from variations in cell-cycle parameters and apoptosis is a crucial step toward better informing drug administration. Antimitotic agents, widely used in chemotherapy, target exclusively proliferative cells and commonly induce a prolonged mitotic arrest followed by cell death via apoptosis. In this paper, we developed a physiologically motivated mathematical framework for describing cancer cell growth dynamics that incorporates the intrinsic heterogeneity in the time individual cells spend in the cell-cycle and apoptosis process. More precisely, our model comprises two age-structured partial differential equations for the proliferative and apoptotic cell compartments and one ordinary differential equation for the quiescent compartment. To reflect the intrinsic cell heterogeneity that governs the growth dynamics, proliferative and apoptotic cells are structured in “age,” i.e., the amount of time remaining to be spent in each respective compartment. In our model, we considered an antimitotic drug whose effect on the cellular dynamics is to induce mitotic arrest, extending the average cell-cycle length. The prolonged mitotic arrest induced by the drug can trigger apoptosis if the time a cell will spend in the cell cycle is greater than the mitotic arrest threshold. We studied the drug’s effect on the long-term cancer cell growth dynamics using different durations of prolonged mitotic arrest induced by the drug. Our numerical simulations suggest that at confluence and in the absence of the drug, quiescence is the long-term asymptotic behavior emerging from the cancer cell growth dynamics. This pattern is maintained in the presence of small increases in the average cell-cycle length. However, intermediate increases in cell-cycle length markedly decrease the total number of cells and can drive the cancer population to extinction. Intriguingly, a large “switch-on/switch-off” increase in the average cell-cycle length maintains an active cell population in the long term, with oscillating numbers of proliferative cells and a relatively constant quiescent cell number. PMID:28913178

Sources and transport of anthropogenic radionuclides in the Ob River system, Siberia

NASA Astrophysics Data System (ADS)

Cochran, J. Kirk; Moran, S. Bradley; Fisher, Nicholas S.; Beasley, Thomas M.; Kelley, James M.

2000-06-01

The potential sources of anthropogenic radionuclides to the Ob River system of western Siberia include global stratospheric fallout, tropospheric fallout from atomic weapons tests and releases from production and reprocessing facilities. Samples of water, suspended and bottom sediments collected in 1994 and 1995 have been used to characterize the sources and transport of 137Cs, Pu isotopes, 237Np and 129I through the system. For the radionuclides that associate with particles, isotope ratios provide clues to their sources, providing any geochemical fractionation can be taken into account. Activity ratios of 239,240Pu/ 137Cs in suspended sediments are lower than the global fallout ratio in the Irtysh River before its confluence with the Ob, comparable to fallout in the central reach of the Ob, and greater than the fallout values in the lower Ob and in the Taz River. This pattern mirrors the downriver decrease in dissolved organic carbon (DOC) concentrations. Laboratory adsorption experiments with Ob River sediment and water show that Kd values for Am (and presumably other actinides) are depressed by two orders of magnitude in the presence of Ob DOC concentrations, relative to values measured in DOC-free Ob water. Iodine and cesium Kd values show little or no (less than a factor of 2) dependence on DOC. Mixing plots using plutonium isotope ratios (atom ratios) show that Pu in suspended sediments of the Ob is a mixture of stratospheric global fallout at northern latitudes, tropospheric fallout from the former Soviet Union test site at Semipalatinsk and reprocessing of spent fuel at Tomsk-7. Plutonium from Semipalatinsk is evident in the Irtysh River above its confluence with the Tobal. Suspended sediment samples taken in the Ob above its confluence with the Irtysh indicate the presence of Pu derived from the Tomsk-7 reprocessing facilities. A mixing plot constructed using 237Np/ 239Pu vs. 240Pu/ 239Pu shows similar mixtures of stratospheric and tropospheric fallout, with the likely addition of inputs from reprocessing facilities and reactor operations. As with Pu/Cs ratios, Np/Pu ratios could be modified by differential geochemical behaviors of Np and Pu. Dissolved 129I only weakly interacts with particles in the Ob; size-fractionated sampling shows that the colloidal 129I fraction (defined as 1 kDa-0.2 μm) contains generally <5% relative to that passing a 0.2 μm filter. Iodine-129 concentrations decrease from 8.3×10 9 to 0.65×10 9 atoms l -1 through the Ob system toward the Kara Sea, with highest values in the Tobal River and lowest in the Taz River. The likely source of the elevated 129I in the Tobal is release from the production-reprocessing facilities at Mayak, and decreases downriver are predominantly due to dilution as the various tributaries with low 129I join the system. Fluxes of 129I to the lower Ob at Salekhard are <1% of the releases of this radionuclide from the nuclear fuel reprocessing facilities at Sellafield, UK and La Hague, France.

CFD Study of Turbo-Ramjet Interactions in Hypersonic Airbreathing Propulsion System

NASA Technical Reports Server (NTRS)

Chang, Ing; Hunter, Louis G.

1996-01-01

Advanced airbreathing propulsion systems used in Mach 4-6 mission scenarios, usually involve turbo-ramjet configurations. As the engines transition from turbojet to ramjet, there is an operational envelope where both engines operate simultaneously. In the first phase of our study, an over/under nozzle configuration was analyzed. The two plumes from the turbojet and ramjet interact at the end of a common 2-D cowl, where they both reach an approximate Mach 3.0 condition and then jointly expand to Mach 3.6 at the common nozzle exit plane. For the problem analyzed, the turbojet engine operates at a higher nozzle pressure ratio than the ramjet, causes the turbojet plume overpowers the ramjet plume, deflecting it approximately 12 degrees downward and in turn the turbojet plume is deflected 6 degrees upward. In the process, shocks were formed at the deflections and a shear layer formed at the confluence of the two jets. This particular case was experimentally tested and the data were used to compare with a computational fluid dynamics (CFD) study using the PARC2D code. The CFD results were in good agreement with both static pressure distributions on the cowl separator and on nozzle walls. The thrust coefficients were also in reasonable agreement. In addition, inviscid relationships were developed around the confluence point, where the two exhaust jets meet, and these results compared favorably with the CFD results. In the second phase of our study, a 3-D CFD solution was generated to compare with the 2-D solution. The major difference between the 2-D and 3-D solutions was the interaction of the shock waves, generated by the plume interactions, on the sidewall. When a shock wave interacts with a sidewall and sidewall boundary layer, it is called a glancing shock sidewall interaction. These interactions entrain boundary layer flow down the shockline into a vortical flow pattern. The 3-D plots show the streamlines being entrained down the shockline. The pressure of the flow also decreases slightly as the sidewall is approached. Other difference between the 2-D and 3-D solutions were a lowering of the nozzle thrust coefficient value from 0.9850 (2-D) to 0.9807 (3-D), where the experimental value was 0.9790. In the third phase of our study, a different turbo-ramjet configuration was analyzed. The confluence of a supersonic turbojet and a subsonic ramjet in the turbine based combined-cycle (TBCC) propulsion system was studied by a 2-D CFD code. In the analysis, Mach 1.4 primary turbojet was mixed with the subsonic ramjet secondary flow in an ejector mode operation. Reasonable agreements were obtained with the supplied I-D TBCC solutions. For low downstream backpressure, the Fabri choke condition (Break-Point condition) was observed in the secondary flow within mixing zone. For sufficient high downstream backpressure, the Fabri choke no longer exist, the ramjet flow was reduced and the ejector flow became backpressure dependent. Highly non-uniform flow at ejector exit were observed, indicated that for smooth downstream combustion, the mixing of the two streams probably required some physical devices.

Bathymetrically controlled velocity-shear front at a tidal river confluence

NASA Astrophysics Data System (ADS)

Blain, Cheryl Ann; Mied, Richard P.; McKay, Paul; Chen, Wei; Rhea, W. Joseph

2015-08-01

Nonbuoyant front formation at the confluence of Nanjemoy Creek and the main Potomac River (MD) channel is examined. Terra satellite ASTER imagery reveals a sediment color front emerging from Nanjemoy Creek when the Potomac is near maximum ebb. Nearly contemporaneous ASTER and Landsat ETM+ imagery are used to extract surface velocities, which suggest a velocity shear front is collocated with the color front. In situ velocities (measured by RiverRay traverses near the Nanjemoy Creek mouth) confirm the shear front's presence. A finite-element simulation (using ADCIRC) replicates the observed velocity-shear front and is applied to decipher its physics. Three results emerge: (1) the velocity-shear front forms, confined to a shoal downstream of the creek-river confluence for most of the tidal cycle, (2) a simulation with a flat bottom in Nanjemoy Creek and Potomac River (i.e., no bathymetry variation) indicates the velocity-shear front never forms, hence the front cannot exist without the bathymetry, and (3) an additional simulation with a blocked-off Creek entrance demonstrates that while the magnitude of the velocity shear is largely unchanged without the creek, shear front formation is delayed in time. Without the Creek, there is no advection of the M6 tidal constituent (generated by nonlinear interaction of the flow with bottom friction) onto the shoals, only a locally generated contribution. A tidal phase difference between Nanjemoy and Potomac causes the ebbing Nanjemoy Creek waters to intrude into the Potomac as far south as its deep channel, and draw from a similar location in the Potomac during Nanjemoy flood.

Earth observations taken by the Expedition 12 crew

NASA Image and Video Library

2005-11-15

ISS012-E-11779 (8 Dec. 2005) --- Ciudad Guayana, Venezuela is featured in this image photographed by an Expedition 12 crew member on the International Space Station. Ciudad Guayana lies on the south bank of the Orinoco River, the second largest in South America, at its confluence with the Caroní (lower left) River. Islands have developed in the Orinoco at a wide sector opposite the city. Guayana is one of the newest cities in Venezuela, having been constructed in 1961 as a new economic growth center for the interior of Venezuela, especially for major industries such as iron, steel, and aluminum. The city stretches 40 kilometers along the south bank of the Orinoco with a population of approximately 800,000 people, one of Venezuela’s largest urban populations. A low barrage (lower left) dams the Caroní as a water supply for the city. The Orinoco River carries brown muddy sediment produced by erosion of the young Andes Mountains far to the southwest. By contrast, the Caroní drains the ancient landscapes of the Guyana Highlands where erosion is so slow that the river water is clear (appearing dark blue) due to a significantly lower sediment load. As at the famous confluence of the brown Amazon and clear Negro rivers in Amazonia, a mixing zone can be seen in the middle of the view, the clear Caroní water producing a less muddy zone for tens of kilometers downstream of the confluence (this part of the Orinoco flows almost due east, left to right).

Ligation of superior mesenteric vein and portal to splenic vein anastomosis after superior mesenteric-portal vein confluence resection during pancreaticoduodenectomy - Case report.

PubMed

Tang, Jianlin; Abbas, Jihad; Hoetzl, Katherine; Allison, David; Osman, Mahamed; Williams, Mallory; Zelenock, Gerald B

2014-12-01

62 year old Caucasian female with pancreatic head mass abutting the superior mesenteric vein (SMV) presented with fine needle aspiration biopsy confirmed diagnosis of ductal adenocarcinoma. CT scan showed near complete obstruction of portal vein and large SMV collateral development. After 3 months of neoadjuvant therapy, her portal vein flow improved significantly, SMV collateral circulation was diminished. Pancreaticoduodenectomy (PD) and superior mesenteric portal vein (SMPV) confluence resection were performed; A saphenous vein interposition graft thrombosed immediately. The splenic vein remnant was distended and adjacent to the stump of the portal vein. Harvesting an internal jugular vein graft required extra time and using a synthetic graft posed a risk of graft thrombosis or infection. As a result, we chose to perform a direct anastomosis of the portal and splenic vein in a desperate situation. The anastomosis decompressed the mesenteric venous system, so we then ligated the SMV. The patient had an uneventful postoperative course, except transient ascites. She redeveloped ascites more than one year later. At that time a PET scan showed bilateral lung and right femur metastatic disease. She expired 15 months after PD. The lessons we learned are (1) Before SMPV confluence resection, internal jugular vein graft should be ready for reconstruction. (2) Synthetic graft is an alternative for internal jugular vein graft. (3) Direct portal vein to SMV anastomosis can be achieved by mobilizing liver. (4) It is possible that venous collaterals secondary to SMV tumor obstruction may have allowed this patient's post-operative survival.

Ligation of superior mesenteric vein and portal to splenic vein anastomosis after superior mesenteric-portal vein confluence resection during pancreaticoduodenectomy – Case report

PubMed Central

Tang, Jianlin; Abbas, Jihad; Hoetzl, Katherine; Allison, David; Osman, Mahamed; Williams, Mallory; Zelenock, Gerald B.

2014-01-01

62 year old Caucasian female with pancreatic head mass abutting the superior mesenteric vein (SMV) presented with fine needle aspiration biopsy confirmed diagnosis of ductal adenocarcinoma. CT scan showed near complete obstruction of portal vein and large SMV collateral development. After 3 months of neoadjuvant therapy, her portal vein flow improved significantly, SMV collateral circulation was diminished. Pancreaticoduodenectomy (PD) and superior mesenteric portal vein (SMPV) confluence resection were performed; A saphenous vein interposition graft thrombosed immediately. The splenic vein remnant was distended and adjacent to the stump of the portal vein. Harvesting an internal jugular vein graft required extra time and using a synthetic graft posed a risk of graft thrombosis or infection. As a result, we chose to perform a direct anastomosis of the portal and splenic vein in a desperate situation. The anastomosis decompressed the mesenteric venous system, so we then ligated the SMV. The patient had an uneventful postoperative course, except transient ascites. She redeveloped ascites more than one year later. At that time a PET scan showed bilateral lung and right femur metastatic disease. She expired 15 months after PD. Conclusion The lessons we learned are (1) Before SMPV confluence resection, internal jugular vein graft should be ready for reconstruction. (2) Synthetic graft is an alternative for internal jugular vein graft. (3) Direct portal vein to SMV anastomosis can be achieved by mobilizing liver. (4) It is possible that venous collaterals secondary to SMV tumor obstruction may have allowed this patient's post-operative survival. PMID:25568802

Discontinuities concentrate mobile predators: Quantifying organism-environment interactions at a seascape scale

USGS Publications Warehouse

Kennedy, Christina G.; Mather, Martha E.; Smith, Joseph M.; Finn, John T.; Deegan, Linda A.

2016-01-01

Understanding environmental drivers of spatial patterns is an enduring ecological problem that is critical for effective biological conservation. Discontinuities (ecologically meaningful habitat breaks), both naturally occurring (e.g., river confluence, forest edge, drop-off) and anthropogenic (e.g., dams, roads), can influence the distribution of highly mobile organisms that have land- or seascape scale ranges. A geomorphic discontinuity framework, expanded to include ecological patterns, provides a way to incorporate important but irregularly distributed physical features into organism–environment relationships. Here, we test if migratory striped bass (Morone saxatilis) are consistently concentrated by spatial discontinuities and why. We quantified the distribution of 50 acoustically tagged striped bass at 40 sites within Plum Island Estuary, Massachusetts during four-monthly surveys relative to four physical discontinuities (sandbar, confluence, channel network, drop-off), one continuous physical feature (depth variation), and a geographic location variable (region). Despite moving throughout the estuary, striped bass were consistently clustered in the middle geographic region at sites with high sandbar area, close to channel networks, adjacent to complex confluences, with intermediate levels of bottom unevenness, and medium sized drop-offs. In addition, the highest striped bass concentrations occurred at sites with the greatest additive physical heterogeneity (i.e., where multiple discontinuities co-occurred). The need to incorporate irregularly distributed features in organism–environment relationships will increase as high-quality telemetry and GIS data accumulate for mobile organisms. The spatially explicit approach we used to address this challenge can aid both researchers who seek to understand the impact of predators on ecosystems and resource managers who require new approaches for biological conservation.

Unit 3, STA. 158+ 40 RB, Hinckson Run culvertdetail ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 3, STA. 158+ 40 RB, Hinckson Run culvert-detail - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 1, STA. 43 + 00 + lb sideslope without ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 1, STA. 43 + 00 + lb sideslope without walls-context - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

26. DETAIL VIEW OF THE ADJUSTABLE FIELD RESISTTOR MANUAL CONTROLS ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

26. DETAIL VIEW OF THE ADJUSTABLE FIELD RESISTTOR MANUAL CONTROLS AND SWITCH BOXES. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

18. TURBINE AND GENERATOR SHAFT IN CONCRETE HOUSING OF THE ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

18. TURBINE AND GENERATOR SHAFT IN CONCRETE HOUSING OF THE TURBINE FLUME. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

8. VIEW OF WESTERN END OF THE POWER PLANT BUILDING ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

8. VIEW OF WESTERN END OF THE POWER PLANT BUILDING LOOKING NORTH. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

Implications of Weltanschauungen for Value Formulation by Guidance Personnel

ERIC Educational Resources Information Center

Weinstock, Henry R.; O'Dowd, Peter S.

1970-01-01

Examines two divergent world-views (empiricism and ration alism) of human values, presenting definitions from many philosophers. Anticipates possibility of confluence of beliefs on nature of man which may offer guidance personnel more accurate tools for direction. (CJ)

3. View Looking Northwest Showing Base of Derrick and Scattered ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

3. View Looking Northwest Showing Base of Derrick and Scattered Pipe Fragments in Foreground - David Renfrew Oil Rig, East side of Connoquenessing Creek, 0.4 mile North of confluence with Thorn Creek, Renfrew, Butler County, PA

4. Band Wheel and Walking Beam Mechanism, Including Remains of ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

4. Band Wheel and Walking Beam Mechanism, Including Remains of Frame Belt House, Looking Southeast - David Renfrew Oil Rig, East side of Connoquenessing Creek, 0.4 mile North of confluence with Thorn Creek, Renfrew, Butler County, PA

15. Detail of Well Head Showing Horizontal Release Pipe for ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

15. Detail of Well Head Showing Horizontal Release Pipe for Natural Gas, Looking North - David Renfrew Oil Rig, East side of Connoquenessing Creek, 0.4 mile North of confluence with Thorn Creek, Renfrew, Butler County, PA

CAT-1 as a novel CAM stabilizes endothelial integrity and mediates the protective actions of L-Arg via a NO-independent mechanism.

PubMed

Guo, Lu; Tian, Shuang; Chen, Yuguo; Mao, Yun; Cui, Sumei; Hu, Aihua; Zhang, Jianliang; Xia, Shen-Ling; Su, Yunchao; Du, Jie; Block, Edward R; Wang, Xing Li; Cui, Zhaoqiang

2015-10-01

Interendothelial junctions play an important role in the maintenance of endothelial integrity and the regulation of vascular functions. We report here that cationic amino acid transporter-1 (CAT-1) is a novel interendothelial cell adhesion molecule (CAM). We identified that CAT-1 protein localized at cell-cell adhesive junctions, similar to the classic CAM of VE-cadherin, and knockdown of CAT-1 with siRNA led to an increase in endothelial permeability. In addition, CAT-1 formed a cis-homo-dimer and showed Ca(2+)-dependent trans-homo-interaction to cause homophilic cell-cell adhesion. Co-immunoprecipitation assays showed that CAT-1 can associate with β-catenin. Furthermore, we found that the sub-cellular localization and function of CAT-1 are associated with cell confluency, in sub-confluent ECs CAT-1 proteins distribute on the entire surface and function as L-Arg transporters, but most of the CAT-1 in the confluent ECs are localized at interendothelial junctions and serve as CAMs. Further functional characterization has disclosed that extracellular L-Arg exposure stabilizes endothelial integrity via abating the cell junction disassembly of CAT-1 and blocking the cellular membrane CAT-1 internalization, which provides the new mechanisms for L-Arg paradox and trans-stimulation of cationic amino acid transport system (CAAT). These results suggest that CAT-1 is a novel CAM that directly regulates endothelial integrity and mediates the protective actions of L-Arg to endothelium via a NO-independent mechanism. Copyright © 2015 Elsevier Ltd. All rights reserved.

The recovery time course of the endothelial-cell glycocalyx in vivo and its implications in vitro

PubMed Central

Potter, Daniel R.; Jiang, John; Damiano, Edward R.

2009-01-01

Compelling evidence continues to emerge suggesting that the glycocalyx surface layer on vascular endothelial cells plays a determining role in numerous physiological processes including inflammation, microvascular permeability, and endothelial mechanotransduction. Previous research has shown that enzymes degrade the glycocalyx, while inflammation causes shedding of the layer. To track the endogenous recovery of the glycocalyx in vivo, we used fluorescent micro-particle image velocimetry (µ-PIV) in mouse cremaster-muscle venules to estimate the hydrodynamically relevant glycocalyx thickness 1, 3, 5, and 7 days after enzymatic or cytokine-mediated degradation of the layer. Results indicate that after acute degradation of the glycocalyx, 5–7 days are required for the layer to endogenously restore itself to its native hydrodynamically relevant thickness in vivo. In light of these findings, and since demonstrable evidence has emerged that standard cell-culture conditions are not conducive to providing the environment and/or cellular conditions necessary to produce and maintain a physiologically relevant cell-surface glycocalyx in vitro, we sought to determine if merely the passage of time would be sufficient to promote the production of a hydrodynamically relevant glycocalyx on a confluent monolayer of human umbilical vein endothelial cells (HUVECs). Using µ-PIV, we found that the hydrodynamically relevant glycocalyx was substantially absent 7 days post-confluence on HUVEC-lined cylindrical collagen microchannels maintained under standard culture conditions. Thus it remains to be determined how a hydrodynamically relevant glycocalyx surface layer can be synthesized and maintained in culture before the endothelial-cell culture model can be used to elucidate glycocalyx-mediated mechanisms of endothelial-cell function. PMID:19443840

Mechanical stimuli differentially control stem cell behavior: morphology, proliferation, and differentiation

PubMed Central

Maul, Timothy M.; Chew, Douglas W.; Nieponice, Alejandro

2011-01-01

Mesenchymal stem cell (MSC) therapy has demonstrated applications in vascular regenerative medicine. Although blood vessels exist in a mechanically dynamic environment, there has been no rigorous, systematic analysis of mechanical stimulation on stem cell differentiation. We hypothesize that mechanical stimuli, relevant to the vasculature, can differentiate MSCs toward smooth muscle (SMCs) and endothelial cells (ECs). This was tested using a unique experimental platform to differentially apply various mechanical stimuli in parallel. Three forces, cyclic stretch, cyclic pressure, and laminar shear stress, were applied independently to mimic several vascular physiologic conditions. Experiments were conducted using subconfluent MSCs for 5 days and demonstrated significant effects on morphology and proliferation depending upon the type, magnitude, frequency, and duration of applied stimulation. We have defined thresholds of cyclic stretch that potentiate SMC protein expression, but did not find EC protein expression under any condition tested. However, a second set of experiments performed at confluence and aimed to elicit the temporal gene expression response of a select magnitude of each stimulus revealed that EC gene expression can be increased with cyclic pressure and shear stress in a cell-contact-dependent manner. Further, these MSCs also appear to express genes from multiple lineages simultaneously which may warrant further investigation into post-transcriptional mechanisms for controlling protein expression. To our knowledge, this is the first systematic examination of the effects of mechanical stimulation on MSCs and has implications for the understanding of stem cell biology, as well as potential bioreactor designs for tissue engineering and cell therapy applications. PMID:21253809

Numerical simulation of transverse mixing of waters at the confluence of two rivers

NASA Astrophysics Data System (ADS)

Lyubimova, Tatyana; Lepikhin, Anatoliy; Parshakova, Yanina

2017-04-01

Surface water bodies, both natural (rivers, lakes) and artificial (ponds, reservoirs) are the main source of drinking water. In this regard, special attention should be paid to their pollution, first of all, to extreme pollution, creating a direct danger to their consumption properties. Traditionally, it is believed that the primary mechanism of the pollutants transport in surface water bodies is the Fick diffusion for which the concentration and nature of the pollutants does not affect the transport mechanisms. However, recent studies have shown that these traditional concepts are not always acceptable. In some cases, even relatively low concentrations of pollutants can fundamentally change the hydrodynamics of a flow through a change in density. Density effects playing in these cases an important role can be very important in solving applied problems of water consumption. One an important question is the mechanism of the formation of the streams do not mixing at large distances from the place of confluence of two rivers with significantly different physical and chemical properties of water. In particular, the Vishera river and the Kama river in Perm Region, as evidenced by the numerous space- and aero photographs, virtually do not mix from the place of their confluence to the territory of Solikamsk-Berezniki industrial area. This is fundamentally important for solving the problem of regulation of technological impacts of this source of technogenic pollution, largest in the basin of the Kama river. In the present paper, the phenomenon of a significant weakening of the transverse mixing of water masses in the surface water bodies is investigated with the help of the numerical simulation of the transverse mixing of water masses at the confluence of two rivers. The calculations are carried out with the help of CFD package ANSYS Fluent using k-epsilon model to describe the turbulent pulsations. The dependence of the transverse mixing speed on the flow rates in the confluent rivers typical for the various seasonal periods is studied. It is found that with the growth of the flow rates in confluent rivers the intensity of mixing is reduced. The work was supported by the Russian Foundation for Basic Research and the Government of Perm region (grant 16-41-590005).

Pathway of oral absorption of heparin with sodium N-[8-(2-hydroxybenzoyl)amino] caprylate.

PubMed

Malkov, Dmitry; Wang, Huai-Zhen; Dinh, Steven; Gomez-Orellana, Isabel

2002-08-01

The oral bioavailability of heparin is negligible. Recent studies, however, have shown that sodium N-[8-(2-hydroxybenzoyl) amino]caprylate (SNAC) and other N-acylated amino acids enable oral heparin absorption. To investigate the mechanism by which heparin crosses the intestinal epithelium in the presence of SNAC, we have used fluorescence microscopy to follow the transport of heparin across Caco-2 cell monolayers. The experiments were carried out on Caco-2 monolayers and Caco-2 cells grown to confluence on culture dishes, using different concentrations of SNAC. The localization of fluorescently labeled heparin was determined using epi-fluorescence and confocal microscopy. DNA dyes were used to determine the effect of SNAC on the plasma membrane integrity. F-actin was labeled with fluorescent phalloidin to investigate the stability of perijunctional actin rings in the presence of SNAC. Heparin was detected in the cytoplasm only after incubation of the cells with heparin and SNAC. No DNA staining was observed in cells incubated with a DNA dye in the presence of SNAC concentrations at which heparin transport occurred. In addition, no signs of actin redistribution or perijunctional ring disbandment were observed during the transport of heparin. The results indicate that SNAC enables heparin transport across Caco-2 monolayers via the transcellular pathway. Heparin transport in the presence of SNAC is selective and does not involve permeabilization of the plasma membrane or tight junction disruption.

Effects of Iron on Physical and Mechanical Properties, and Osteoblast Cell Interaction in β-Tricalcium Phosphate

PubMed Central

Vahabzadeh, Sahar; Bose, Susmita

2017-01-01

Iron (Fe) is a vital element and its deficiency causes abnormal bone metabolism. We investigated the effects of Fe and its concentration in β-tricalcium phosphate (β-TCP) on physicomechanical properties and in vitro proliferation and differentiation of osteoblasts. Our results showed that Fe addition at concentrations of 0.5 wt. % (0.5 Fe-TCP) and 1.0 wt. % (1.0 Fe-TCP) inhibits the β-TCP to α-TCP phase transformation at sintering temperature of 1250 °C. Addition of 0.25 wt. % Fe (0.25 Fe-TCP) increased the compressive strength of β-TCP from 167.27±16.2 MPa to 227.10±19.3 MPa. After 3 days of culture, surfaces of 0.5 Fe-TCP and 1.0 Fe-TCP samples were covered by osteoblast cells, compared to that of pure and 0.25 Fe-TCP. Cells grew to confluency on all Fe-doped samples after 7 days of culture and monolayer sheetlike cellular structure was found at 11 days. Optical cell density and alkaline phosphatase activity were significantly higher on Fe-doped samples and the highest values were found in 0.5 Fe-TCP samples. Our results show that Fe concentration had significant effect on physical and mechanical properties of TCP ceramics, and also on the in vitro osteoblast cellular interactions in TCP ceramics. PMID:27896489

On the Normal Force Mechanotransduction of Human Umbilical Vein Endothelial Cells

NASA Astrophysics Data System (ADS)

Vahabikashi, Amir; Wang, Qiuyun; Wilson, James; Wu, Qianhong; Vucbmss Team

2016-11-01

In this paper, we report a cellular biomechanics study to examine the normal force mechanotransduction of Human Umbilical Vein Endothelial Cells (HUVECs) with their implications on hypertension. Endothelial cells sense mechanical forces and adjust their structure and function accordingly. The mechanotransduction of normal forces plays a vital role in hypertension due to the higher pressure buildup inside blood vessels. Herein, HUVECs were cultured to full confluency and then exposed to different mechanical loadings using a novel microfluidic flow chamber. One various pressure levels while keeps the shear stress constant inside the flow chamber. Three groups of cells were examined, the control group (neither shear nor normal stresses), the normal pressure group (10 dyne/cm2 of shear stress and 95 mmHg of pressure), and the hypertensive group (10 dyne/cm2 of shear stress and 142 mmHg of pressure). Cellular response characterized by RT-PCR method indicates that, COX-2 expressed under normal pressure but not high pressure; Mn-SOD expressed under both normal and high pressure while this response was stronger for normal pressure; FOS and e-NOS did not respond under any condition. The differential behavior of COX-2 and Mn-SOD in response to changes in pressure, is instrumental for better understanding the pathogenesis of hypertensive cardiovascular diseases. This research was supported by the National Science Foundation under Award #1511096.

Impedance spectroscopy for the detection and identification of unknown toxins

NASA Astrophysics Data System (ADS)

Riggs, B. C.; Plopper, G. E.; Paluh, J. L.; Phamduy, T. B.; Corr, D. T.; Chrisey, D. B.

2012-06-01

Advancements in biological and chemical warfare has allowed for the creation of novel toxins necessitating a universal, real-time sensor. We have used a function-based biosensor employing impedance spectroscopy using a low current density AC signal over a range of frequencies (62.5 Hz-64 kHz) to measure the electrical impedance of a confluent epithelial cell monolayer at 120 sec intervals. Madin Darby canine kidney (MDCK) epithelial cells were grown to confluence on thin film interdigitated gold electrodes. A stable impedance measurement of 2200 Ω was found after 24 hrs of growth. After exposure to cytotoxins anthrax lethal toxin and etoposide, the impedance decreased in a linear fashion resulting in a 50% drop in impedance over 50hrs showing significant difference from the control sample (~20% decrease). Immunofluorescent imaging showed that apoptosis was induced through the addition of toxins. Similarities of the impedance signal shows that the mechanism of cellular death was the same between ALT and etoposide. A revised equivalent circuit model was employed in order to quantify morphological changes in the cell monolayer such as tight junction integrity and cell surface area coverage. This model showed a faster response to cytotoxin (2 hrs) compared to raw measurements (20 hrs). We demonstrate that herein that impedance spectroscopy of epithelial monolayers serves as a real-time non-destructive sensor for unknown pathogens.

Flood-inundation maps for Sweetwater Creek from above the confluence of Powder Springs Creek to the Interstate 20 bridge, Cobb and Douglas Counties, Georgia

USGS Publications Warehouse

Musser, Jonathan W.

2012-01-01

Digital flood-inundation maps for a 10.5-mile reach of Sweetwater Creek, from about 1,800 feet above the confluence of Powder Springs Creek to about 160 feet below the Interstate 20 bridge, were developed by the U.S. Geological Survey (USGS) in cooperation with Cobb County, Georgia. The inundation maps, which can be accessed through the USGS Flood Inundation Mapping Science Web site at http://water.usgs.gov/osw/flood_inundation/, depict estimates of the areal extent and depth of flooding corresponding to selected water levels (stages) at the USGS streamgage at Sweetwater Creek near Austell, Georgia (02337000). Current stage at this USGS streamgage may be obtained at http://waterdata.usgs.gov/ and can be used in conjunction with these maps to estimate near real-time areas of inundation. The National Weather Service (NWS) is incorporating results from this study into the Advanced Hydrologic Prediction Service (AHPS) flood-warning system (http://water.weather.gov/ahps/). The NWS forecasts flood hydrographs at many places that commonly are collocated at USGS streamgages. The forecasted peak-stage information for the USGS streamgage at Sweetwater Creek near Austell (02337000), which is available through the AHPS Web site, may be used in conjunction with the maps developed in this study to show predicted areas of flood inundation. A one-dimensional step-backwater model was developed using the U.S. Army Corps of Engineers Hydrologic Engineering Centers River Analysis System (HEC–RAS) software for Sweetwater Creek and was used to compute flood profiles for a 10.5-mile reach of the creek. The model was calibrated using the most current stage-discharge relations at the Sweetwater Creek near Austell streamgage (02337000), as well as high-water marks collected during annual peak-flow events in 1982 and 2009. The hydraulic model was then used to determine 21 water-surface profiles for flood stages at the Sweetwater Creek streamgage at 1-foot intervals referenced to the streamgage datum and ranging from just above bankfull stage (12.0 feet) to approximately 1.2 feet above the highest recorded water level at the streamgage (32.0 feet). The simulated water-surface profiles were then combined with a geographic information system digital elevation model—derived from contour data (8-foot horizontal resolution), in Cobb County, and USGS National Elevation Dataset (31-foot horizontal resolution), in Douglas County—to delineate the area flooded for each 1-foot increment of stream stage. The availability of these maps, when combined with real-time information regarding current stage from USGS streamgages and forecasted stream stages from the NWS, provides emergency management personnel and residents with critical information during flood-response activities, such as evacuations and road closures, as well as for post-flood recovery efforts.

Radiologic evaluation of vasculobiliary anatomy in the umbilical fissure.

PubMed

Ji, Gu-Wei; Zhu, Fei-Peng; Wang, Ke; Xia, Yong-Xiang; Jiao, Chen-Yu; Shao, Zi-Cheng; Li, Xiang-Cheng

2017-06-15

Preoperative evaluation of vasculobiliary anatomy in the umbilical fissure (U-point) is pivotal for perihilar cholangiocarcinoma (PCCA) applied to right-sided hepatectomy. The purpose of our study was to review the vasculobiliary anatomy in the U-point using three-dimensional (3D) reconstruction technique, to investigate the diagnostic ability of 2D scans to evaluate anatomic variations, and to discuss its surgical implications. A retrospective study of 159 patients with Bismuth type I, II, and IIIa PCCA, who received surgery at our institution from November 2012 to September 2016, was conducted. Anatomic structures were assessed using multidetector computed tomography (MDCT) by one hepatobiliary surgeon, whereas 3D images were reconstructed by an independent radiologist. Normal confluence pattern of left biliary system was defined as the left medial segmental bile duct (B4) joining the common trunk of segment II (B2) and segment III (B3) ducts, whereas aberrant confluence patterns were classified into 3 types: type I, triple confluence of B2, B3, and B4; type II, B2 draining into the common trunk of B3 and B4; type III, other patterns. Surgical anatomy of B4 was classified into the central, peripheral, and combined type according to its relation to the hepatic confluence. The lengths from the bile duct branch of Spiegel's lobe (B1l) to the orifice of B4 and the junction of B2 and B3 were measured on 3D images. The anatomy of left hepatic artery (LHA) was classified according to different origins and the spatial relationship related to the U-point. 3D reconstruction revealed that normal confluence pattern of left biliary system was observed in 71.1% (113/159) of all patients, and variant patterns were type I in 11.9% (19/159), type II in 12.6% (20/159), and type III in 4.4% (7/159). The length from B1l to the junction of B2 and B3 was 12.1 ± 3.1 mm in type I variation, which was significantly shorter than that in normal configuration (30.0 ± 6.8 mm, P < 0.001) but significantly longer than that in type II variation (9.6 ± 3.4 mm, P = 0.019). Surgical anatomy of B4: the peripheral type was most commonly seen (74.2%, 118/159), followed by central type (15.7%, 25/159) and combined type (10.1%, 16/159). The distance between the B1l and B4 was 8.4 ± 2.4 mm in central and combined type, which was significantly shorter than that in peripheral type (14.5 ± 4.1 mm, P < 0.001). A replaced or accessory LHA from the left gastric artery was present in 6 (3.8%) and 9 (5.7%) patients, respectively. LHA running along the left caudal position of U-point was present in 143 cases (89.9%), along the right cranial position of U-point in nine cases (5.7 %), and combined position in seven cases (4.4%). Interobserver agreement of two imaging modalities was almost perfect in biliary confluence pattern (kappa = 0.90; 95% confidence interval: 0.79-1.00), substantial in surgical anatomy of B4 (kappa = 0.74; 95% confidence interval: 0.62-0.86), and perfect in LHA (kappa = 1.00). Thoroughly understanding the imaging characters of surgical anatomy in the U-point may be benefit for preoperative evaluation of PCCA by successive review of 2D images alone, whereas 3D reconstruction technique allows detailed hepatic anatomy and individualized surgical planning for advanced cases. Copyright © 2017 Elsevier Inc. All rights reserved.

Magnetic nanomaterials and sensors for biological detection.

PubMed

Sobczak-Kupiec, Agnieszka; Venkatesan, Jayachandran; Alhathal AlAnezi, Adnan; Walczyk, Dorota; Farooqi, Ammad; Malina, Dagmara; Hosseini, Seyed Hossein; Tyliszczak, Bozena

2016-11-01

It is becoming progressively more understandable that sensitivity and versatility of magnetic biosensors provides unique platform for high performance diagnostics in clinical settings. Confluence of information suggested that magnetic biosensors required well-tailored magnetic particles as probes for detection that generate large and specific biological signal with minimum possible nonspecific binding. However, there are visible knowledge gaps in our understanding of the strategies to overcome existing challenges related to even smaller size of intracellular targets and lower signal-to-noise ratio than that in whole-cell studies, therefore tool designing and development for intracellular measurement and manipulation is problematic. In this review we describe magnetic nanoparticles, synthesis and sensing principles of magnetic nanoparticles as well as surface functionalization and modification and finally magnetic nanoparticles for medical diagnostics. This review gathers important and up-to-date information and may help to develop the method of obtaining magnetic materials especially for medical application. Copyright © 2016 Elsevier Inc. All rights reserved.

Bacterial colonization stimulates a complex physiological response in the immature human intestinal epithelium

PubMed Central

Hill, David R; Huang, Sha; Nagy, Melinda S; Yadagiri, Veda K; Fields, Courtney; Mukherjee, Dishari; Bons, Brooke; Dedhia, Priya H; Chin, Alana M; Tsai, Yu-Hwai; Thodla, Shrikar; Schmidt, Thomas M; Walk, Seth

2017-01-01

The human gastrointestinal tract is immature at birth, yet must adapt to dramatic changes such as oral nutrition and microbial colonization. The confluence of these factors can lead to severe inflammatory disease in premature infants; however, investigating complex environment-host interactions is difficult due to limited access to immature human tissue. Here, we demonstrate that the epithelium of human pluripotent stem-cell-derived human intestinal organoids is globally similar to the immature human epithelium and we utilize HIOs to investigate complex host-microbe interactions in this naive epithelium. Our findings demonstrate that the immature epithelium is intrinsically capable of establishing a stable host-microbe symbiosis. Microbial colonization leads to complex contact and hypoxia driven responses resulting in increased antimicrobial peptide production, maturation of the mucus layer, and improved barrier function. These studies lay the groundwork for an improved mechanistic understanding of how colonization influences development of the immature human intestine. PMID:29110754

The Activity of Differentiation Factors Induces Apoptosis in Polyomavirus Large T-Expressing Myoblasts

PubMed Central

Fimia, Gian Maria; Gottifredi, Vanesa; Bellei, Barbara; Ricciardi, Maria Rosaria; Tafuri, Agostino; Amati, Paolo; Maione, Rossella

1998-01-01

It is commonly accepted that pathways that regulate proliferation/differentiation processes, if altered in their normal interplay, can lead to the induction of programmed cell death. In a previous work we reported that Polyoma virus Large Tumor antigen (PyLT) interferes with in vitro terminal differentiation of skeletal myoblasts by binding and inactivating the retinoblastoma antioncogene product. This inhibition occurs after the activation of some early steps of the myogenic program. In the present work we report that myoblasts expressing wild-type PyLT, when subjected to differentiation stimuli, undergo cell death and that this cell death can be defined as apoptosis. Apoptosis in PyLT-expressing myoblasts starts after growth factors removal, is promoted by cell confluence, and is temporally correlated with the expression of early markers of myogenic differentiation. The block of the initial events of myogenesis by transforming growth factor β or basic fibroblast growth factor prevents PyLT-induced apoptosis, while the acceleration of this process by the overexpression of the muscle-regulatory factor MyoD further increases cell death in this system. MyoD can induce PyLT-expressing myoblasts to accumulate RB, p21, and muscle- specific genes but is unable to induce G00 arrest. Several markers of different phases of the cell cycle, such as cyclin A, cdk-2, and cdc-2, fail to be down-regulated, indicating the occurrence of cell cycle progression. It has been frequently suggested that apoptosis can result from an unbalanced cell cycle progression in the presence of a contrasting signal, such as growth factor deprivation. Our data involve differentiation pathways, as a further contrasting signal, in the generation of this conflict during myoblast cell apoptosis. PMID:9614186

The effects of blood and blood products on the arachnoid cell.

PubMed

Hansen, Eric A; Romanova, Liudmila; Janson, Christopher; Lam, Cornelius H

2017-06-01

After traumatic brain injury (TBI), large amounts of red blood cells and hemolytic products are deposited intracranially creating debris in the cerebrospinal fluid (CSF). This debris, which includes heme and bilirubin, is cleared via the arachnoid granulations and lymphatic systems. However, the mechanisms by which erythrocytes and their breakdown products interfere with normal CSF dynamics remain poorly defined. The purpose of this study was to model in vitro how blood breakdown products affect arachnoid cells at the CSF-blood barrier, and the extent to which the resorption of CSF into the venous drainage system is mechanically impaired following TBI. Arachnoid cells were grown to confluency on permeable membranes. Rates of growth and apoptosis were measured in the presence of blood and lysed blood, changes in transepithelial electrical resistance (TEER) was measured in the presence of blood and hemoglobin, and small molecule permeability was determined in the presence of blood, lysed blood, bilirubin, and biliverdin. These results were directly compared with an established rat brain endothelial cell line (RBEC4) co-cultured with rat brain astrocytes. We found that arachnoid cells grown in the presence of whole or lysed erythrocytes had significantly slower growth rates than controls. Bilirubin and biliverdin, despite their low solubilities, altered the paracellular transport of arachnoid cells more than the acute blood breakdown components of whole and lysed blood. Mannitol permeability was up to four times higher in biliverdin treatments than controls, and arachnoid membranes demonstrated significantly decreased small molecule permeabilities in the presence of whole and lysed blood. We conclude that short-term (<24 h) arachnoid cell transport and long-term (>5 days) arachnoid cell viability are affected by blood and blood breakdown products, with important consequences for CSF flow and blood clearance after TBI.

A Study of Parameters Affecting Fibroblast Morphology in Response to an Applied Mechanical Force

NASA Technical Reports Server (NTRS)

Grymes, Rosalind A.; Sawyer, Christine

1994-01-01

A precisely controlled stretch/relaxation regimen (20% elongation at 6.6 cycles/min) was applied to normal human fetal, neonatal and aged dermal fibroblasts cultured on flexible membranes. Culture conditions included poly (NH2) or collagen type I coated substrate membranes; control cultures were grown on the same pliable material in the absence of applied stretch. Direct observation and immunofluorescence analyses revealed a progressive change in cell body orientation limited to the stretched dermal fibroblast cultures. Monolayers gradually (over 4 days) acquired a symmetric, radial distribution equivalent to the biaxial array of the applied force. At high seeding density, alignment was inhibited in the fetal cell cultures. This cell strain required collagen type I coating for optimal attachment to the flexible membrane, preferring growth in three-dimensional cell 'balls' on the poly(NH2) coated substrate. Neonatal cells also required the collagen type I coating, but both neonatal and aged dermal fibroblasts aligned efficiently at all seeding densities examined. The randomly oriented neonatal cells on the unstretched control membranes spontaneously detached at confluence, as a single cell sheet. Their aligned counterparts did not detach until the applied stretch stimulus was removed. Low concentrations of cytochalasin D (62.5 ng/ml) disrupted the stretch-related alignment response. Rhodamine phalloidin staining visualized fewer actin stress fibers in stretched, aligned cells than in controls. Both intercellular interactions and cytoskeletal integrity mediate the response to mechanical strain. Normal rabbit corneal stroma fibroblasts (NRC) were also analyzed, and failed to orient under these conditions. This cell type may require a different regimen, or a longer time period, to demonstrate alignment behavior. Supported by NASA Space Biology RTOP 199-40-22 and the NASA-ARC Director's Discretionary Fund.

Organizational behavior of human umbilical vein endothelial cells

PubMed Central

1982-01-01

Culture conditions that favor rapid multiplication of human umbilical vein endothelial cells (HUV-EC) also support long-term serial propagation of the cells. This is routinely achieved when HUV-EC are grown in Medium 199 (M-199) supplemented with fetal bovine serum (FBS) and endothelial cell growth factor (ECGF), on a human fibronectin (HFN) matrix. The HUV-EC can shift from a proliferative to an organized state when the in vitro conditions are changed from those favoring low density proliferation to those supporting high density survival. When ECGF and HFN are omitted, cultures fail to achieve confluence beyond the first or second passage: the preconfluent cultures organize into tubular structures after 4-6 wk. Some tubes become grossly visible and float in the culture medium, remaining tethered to the plastic dish at either end of the tube. On an ultrastructural level, the tubes consist of cells, held together by junctional complexes, arranged so as to form a lumen. The smallest lumens are formed by one cell folding over to form a junction with itself. The cells contain Weibel-Palade bodies and factor VIII-related antigen. The lumens contain granular, fibrillar and amorphous debris. Predigesting the HFN matrix with trypsin (10 min, 37 degrees C) or plasmin significantly accelerates tube formation. Thrombin and plasminogen activator had no apparent effect. Disruption of the largest tubes with trypsin/EDTA permits the cells to revert to a proliferative state if plated on HFN, in M-199, FBS, and ECGF. These observations indicate that culture conditions that do not favor proliferation permit attainment of a state of nonterminal differentiation (organization) by the endothelial cell. Furthermore, proteolytic modification of the HFN matrix may play an important role in endothelial organization. PMID:6813338

CDB-4124 does not cause apoptosis in cultured fibroid cells.

PubMed

Roeder, Hilary; Jayes, Friederike; Feng, Liping; Leppert, Phyllis C

2011-09-01

Selective progesterone receptor modulators (SPRMs), such as asoprisnil (J867) and ulipristal (CDB-2914), have been shown to reduce fibroid volume in vivo and to induce apoptosis in vitro. CDB-4124 (telapristone), a SPRM with different side groups, also reduced fibroid volume in vivo, and we hypothesized that this SPRM would also cause apoptosis in cultured fibroid cells. Immortalized, progesterone receptor-positive fibroid cells, known to be capable of apoptosis, were grown to 80% confluence in serum-containing media. Cells were then treated for 48 hours in serum-free media with 0, 10, 100, or 1000 nmol/L CDB-4124. Actinomycin-D and staurosporine were used as positive controls to induce apoptosis. Apoptosis was quantified using a TUNEL-fluorescein kit. Images were captured with a widefield-fluorescence microscope and analyzed using MetaMorph image analysis software. To validate results, Western blots of total cell lysates were probed for cleaved caspase-3 (c-CASP3). Experiments were repeated 3 times using independent cell batches. Analysis of 19 712 nuclei indicated 14.8% ± 10.9% (mean ± SEM), 8.4% ± 4.6%, 8.2% ± 4.7%, and 9.3% ± 6.3% apoptosis in 0, 10, 100, and 1000 nmol/L CDB-4124-treated cells, respectively. There was no evidence of elevated c-CASP3 over vehicle control after treatment with CDB-4124. CDB-4124 did not significantly induce apoptosis in cultured fibroid cells under the conditions described suggesting apoptosis may not be the main pathway responsible for CDB-4124-induced fibroid shrinkage. Variations in SPRM biological effects may be due to differences in fibroid source cells, binding kinetics, or extracellular matrix characteristics, and can be exploited in further investigations of the mechanisms of action of SPRMs in fibroid biology.

Endoscopic bilateral stent-in-stent placement for malignant hilar obstruction using a large cell type stent.

PubMed

Park, Jin Myung; Lee, Sang Hyub; Chung, Kwang Hyun; Jang, Dong Kee; Ryu, Ji Kon; Kim, Yong-Tae; Lee, Jae Min; Paik, Woo Hyun

2016-12-01

Bilateral stent-in-stent (SIS) self-expandable metal stent placement is technically challenging for palliation of unresectable malignant hilar obstruction. In the SIS technique, the uniform large cell type biliary stent facilitates contralateral stent deployment through the mesh of the first metallic stent. This study aimed to assess the technical success and clinical effectiveness of this technique with a uniform large cell type biliary stent. Thirty-one patients who underwent bilateral SIS placement using a large cell type stent were reviewed retrospectively. All patients showed malignant hilar obstruction (Bismuth types II, III, IV) with different etiologies. Sixteen (51.6%) patients were male. The mean age of the patients was 67.0+/-14.0 years. Most patients were diagnosed as having hilar cholangiocarcinoma (58.1%) and gallbladder cancer (29.0%). Technical success rate was 83.9%. Success was achieved more frequently in patients without masses obstructing the biliary confluence (MOC) than those with MOC (95.2% vs 60.0%, P=0.03). Functional success rate was 77.4%. Complications occurred in 29.0% of the patients. These tended to occur more frequently in patients with MOC (50.0% vs 19.0%, P=0.11). Median time to recurrent biliary obstruction was 188 days and median survival was 175 days. The large cell type stent can be used efficiently for bilateral SIS placement in malignant hilar obstruction. However, the risk of technical failure increases in patients with MOC, and caution is needed to prevent complications for these patients.

An expert protocol for immunofluorescent detection of calcium channels in tsA-201 cells.

PubMed

Koch, Peter; Herzig, Stefan; Matthes, Jan

Pore-forming subunits of voltage gated calcium channels (VGCC) are large membrane proteins (260kDa) containing 24 transmembrane domains. Despite transfection with viral promoter driven vectors, biochemical analysis of VGCC is often hampered by rather low expression levels in heterologous systems rendering VGCC challenging targets. Especially in immunofluorescent detection, calcium channels are demanding proteins. We provide an expert step-by-step protocol with adapted conditions for handling procedures (tsA-201 cell culture, transient transfection, incubation time and temperature at 28°C or 37°C and immunostaining) to address the L-type calcium-channel pore Ca v 1.2 in an immunofluorescent approach. We performed immunocytochemical analysis of Ca v 1.2 expression at single-cell level in combination with detection of different markers for cellular organelles. We show confluency levels and shapes of tsA-201 cells at different time points during an experiment. Our experiments reveal sufficient levels of Ca v 1.2 protein and a correct Ca v 1.2 expression pattern in polygonal shaped cells already 12h after transfection. A sequence of elaborated protocol modifications allows subcellular localization analysis of Ca v 1.2 in an immunocytochemical approach. We provide a protocol that may be used to achieve insights into physiological and pathophysiological processes involving voltage gated calcium channels. Our protocol may be used for expression analysis of other challenging proteins and efficient overexpression may be exploited in related biochemical techniques requiring immunolabels. Copyright © 2016 Elsevier Inc. All rights reserved.

Unit 5, STA. 13+00+RB, point stadiumcontext Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 5, STA. 13+00+RB, point stadium-context - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 5, STA, 13+00+RB, point stadiumdetail Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 5, STA, 13+00+RB, point stadium-detail - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 3, STA. 6+00 Little Conemaugh River, weirdetail Johnstown ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 3, STA. 6+00 Little Conemaugh River, weir-detail - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 5, STA. 19+00+RB, access rampcontext Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 5, STA. 19+00+RB, access ramp-context - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 5, STA. 53+25+RB, Conrad buildingcontext Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 5, STA. 53+25+RB, Conrad building-context - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 3, STA. 158+40 RB, Hinckson Run culvert context ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 3, STA. 158+40 RB, Hinckson Run culvert context - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 3, STA. 6+00 Little Conemaugh River, weircontext Johnstown ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 3, STA. 6+00 Little Conemaugh River, weir-context - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 5, STA. 53+25+RB, Conrad buildingdetail Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 5, STA. 53+25+RB, Conrad building-detail - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

Unit 5, STA. 19+00+RB, access rampdetail Johnstown Local Flood ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Unit 5, STA. 19+00+RB, access ramp-detail - Johnstown Local Flood Protection Project, Beginning on Conemaugh River approx 3.8 miles downstream from confluence of Little Conemaugh & Stony Creek Rivers at Johnstown, Johnstown, Cambria County, PA

6. VIEW OF POWER PLANT BUILDING LOOKING NORTHWEST. DRY CANAL ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

6. VIEW OF POWER PLANT BUILDING LOOKING NORTHWEST. DRY CANAL BED TO THE LEFT. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

9. EXTERIOR PERSPECTIVE OF POWER PLANT BUILDING LOOKING NORTHEAST. DRY ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

9. EXTERIOR PERSPECTIVE OF POWER PLANT BUILDING LOOKING NORTHEAST. DRY CANAL BED IN FOREGROUND. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

1. VIEW OF POWER PLANT LOOKING SOUTHEAST. SEVEN TURBINE FLUMES ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

1. VIEW OF POWER PLANT LOOKING SOUTHEAST. SEVEN TURBINE FLUMES VISIBLE IN FRONT OF BUILDING. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

3. VIEW OF POWER PLANT LOOKING SOUTH INTO THE REMAINS ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

3. VIEW OF POWER PLANT LOOKING SOUTH INTO THE REMAINS OF THE TURBINE FLUMES. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

7. EXTERIOR OF POWER PLANT BUILDING LOOKING NORTHWEST. DETAIL OF ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

7. EXTERIOR OF POWER PLANT BUILDING LOOKING NORTHWEST. DETAIL OF TRASH RACK IN FOREGROUND. - Potomac Power Plant, On West Virginia Shore of Potomac River, about 1 mile upriver from confluence with Shenandoah River, Harpers Ferry, Jefferson County, WV

COMPLETED STRUCTURE. View is eastsoutheast of downstream side of bridge, ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

COMPLETED STRUCTURE. View is east-southeast of downstream side of bridge, from beyond confluence of Trinity and South Fork Trinity Rivers - South Fork Trinity River Bridge, State Highway 299 spanning South Fork Trinity River, Salyer, Trinity County, CA