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Joseph Mitchell
Joseph Mitchell

Tight Mature REPACK



This study demonstrates the presence of tight junction antigens in adult and developing human epidermis. Indirect immunofluorescence labeling and immunoelectron microscopy with antibodies to ZO-1 and occludin localized tight junction components ZO-1 and occludin to a narrow zone of the granular cells of adult epidermis. Double immunolabeling for tight junction components with adherens junction or desmosome proteins suggested that occludin is more specific for tight junctions than ZO-1, which may also be associated with adherens junctions. In developing skin, tight junctions interconnected the peridermal cells, and after the fetal stratification localized to the granular cell layer. Immunolabeling of psoriasis, lichen planus, and ichthyosis vulgaris, representing aberrant differentiation of the epidermis, showed that these conditions were associated with relocation of ZO-1 and occludin to the spinous cells. Cultures of epidermal keratinocytes, which offer a useful model for the formation of cellular contacts, revealed that tight junction components, ZO-1 and occludin, displayed a marked degree of colocalization relatively late during the process when the fusion zone had assumed a linear appearance. This suggests that the formation of adherens junctions and desmosomes precedes that of tight junctions. We speculate that the epidermal barrier, isolating the human body from the external environment, is in part formed by tight junctions of stratum granulosum.




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The Margham Dubai Establishment (MDE) asset is a mature gas-condensate field that was put on production in 1985. As the energy demands of Dubai are ever increasing, MDE has been releasing (net) gas resulting in rapid dwindling of remaining reserves. In order to reduce the amount of alternate fuels required to meet the Dubai energy demands, MDE initiated a drastic field intervention campaign prior to June 1, 2006. Preliminary feasibility studies concluded that the only identified practical option of increasing drainage capacity was to re-enter existing wells drilling high deviation multi-laterals with coiled-tubing, underbalanced drilling (CTUBD) techniques. After approximately three months of rigorous project planning and engineering, the well sequence of Phase I was initiated (4 wells). Upon completion, a 73% net increase (including average field decline) of gas from the Margham field was observed equating to a gas lift cost of less than $2/Mscf (25% less cost than alternate fuels). Due to the success of Phase I, an additional four wells were approved. Upon Phase II completion, a 125% increase (over nonintervention predicted field production rate) resulted, equating to a gas lift cost of less than $1.33/Mscf. The reduction in lift cost in Phase II was attributed to the cost of front-end engineering, mobilization, retention of equipment and additional start-up costs all being absorbed in Phase I. Also, the technical limit (rig move, milling time, drilling time, etc.) for the CTUBD operation was not reached until the drilling of the third well. Thereafter a third and final phase was approved consisting of three additional wells. The total number of wells rejuvenated was eleven.


This paper addresses the conceptualization of the project, the well selection criteria, identified risk and uncertainties, the risk management methods employed, environmental impact, implementation practices, key achievements, production improvement, ultimate recovery enhancement and a cost versus value summary. Overall the CTUBD intervention campaign as a mature field rejuvenation technique has been a great technical and economic achievement. In addition to the apparent increase in production, the rejuvenation of the Margham field will allow for more efficient gas storage methods in the future (if required) due to the increased injectivity index and the ability to use fewer wells for gas cycling.


Colorado also signed two other tight ends in February: Will Pericak (6-4, 245) from Boulder High School and Ryan Wallace (6-5, 230) from Bowling Green, Ky. Each has made some nice catches during their first two days of practice.


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The maturation process of newly-formed subretinal vessels was studied ultrastructurally. Primitive new vessels have thick endothelial cells with cytoplasm that contains many ribosomes and rough endoplasmic reticulum. Some immature endothelial cells show budding, and many of the intercellular junctions have small gaps. As the newly-formed vessels mature, the endothelial cells become attenuated and fenestrated, and their junctions become tight. Mature subretinal new vessels closely resemble those of the normal choriocapillaris.


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I sunk into the cloud of bubbles. I had longish red hair with a natural wave in it and a tight body with pointed, high breasts, a petite stomach and flared hips. I had slept with two men; one was a drunken mistake and the other I had been dating for about two years. They had both been in my age group, but I was curious as to what an older man would be like. That's where my thoughts about Jack came in.


During the movie I was distracted by Jack. I wanted to hold his hand. I liked the way the screen highlighted his face. Occasionally our hands would touch as we would dig into the tub of popcorn and we would exchange a smile. But the popcorn ran out, and now there was no reason to touch. And I wanted to touch him. In the bedroom, I was submissive and wondered what power his charms would work on me. Before I had time to act, he put his arm around me and I rested my head on his shoulder. It seemed so natural. I felt so excited inside. This man was old enough to be my father, but that didn't seem to bother either of us. In all his years, he must've had plenty of women, but if we were ever to sleep together, I wanted to be his greatest and his tightest.


I took off my heels. I knew Jack liked heels. I remember him telling me that once and these particular pumps went well with my jeans and sweater. And this particular sweater was tight and showed everything.


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Cdc42 regulates tight junction formation in 16HBE cells. 16HBE cells were seeded at low density on glass coverslips and transfected with the indicated siRNA. (A) Three days after transfection cells were fixed and analyzed by immunofluorescence microscopy with anti-occludin (green) and Hoechst (blue). Scale bar, 20 μm in all images. (B) Quantification of tight junction formation from three independent experiments (see Materials and Methods for details); error bars, SEM; nsd, no significant difference, **p


PAK4, Par6B, and aPKC regulate tight junction formation in 16HBE cells. (A) 16HBE cells were transfected with the indicated siRNAs. Three days after transfection, cells were fixed and analyzed by immunofluorescence microscopy with anti-ZO-1 (green) and Hoechst (blue). Scale bar, 20 μm in all images. (B, D, and F) Quantification of tight junction formation from three independent experiments after transfection of the indicated siRNAs or treatment with the indicated inhibitors (see Materials and Methods for details); error bars, SEM; nsd, no significant difference, ***p


PAK4 function during tight junction formation is Cdc42- and kinase-dependent. (A) Sequence alignment of the CRIB motif of PAK family kinases with conserved residues in bold. Mutations introduced in to PAK4 to generate PAK4*CRIB are indicated in red. (B) myc-PAK4, but not myc-PAK4*CRIB, coimmunoprecipitates with GFP-Cdc42-L61 when expressed in HEK293T cells. (C and D) Western blot analysis, using the indicated antibodies, of lysates from 16HBE cells described in F. (E) Quantification of tight junction formation from F. Three independent experiments were quantified (see Materials and Methods); error bars, SEM; nsd, no significant difference; **p 041b061a72


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