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To form epithelial organs cells must polarize and generate de novo an apical domain and lumen. Epithelial polarization is masterminded by polarity complexes, which are thought to direct downstream events such as polarized membrane traffic, though this interconnection is not well understood. We report that Rab11a regulates apical traffic and lumen formation via the Rab GEF Rabin8, and its target Rab8a. Rab8a/11a act via the exocyst to target Par3 to the apical surface, and control apical Cdc42 activation via the Cdc42 GEF, Tuba. These components assemble at a transient apical membrane initiation site to form the lumen. This Rab11a-directed network directs Cdc42-dependent apical exocytosis during lumen formation, revealing a novel interplay of the machineries of vesicular transport and polarization.
Most internal epithelial organs consist of a monolayer of polarized epithelial cells surrounding a central lumen. Polarization requires the interaction of the signaling complexes and scaffolds that define cortical domains with the polarized membrane sorting machinery1. In yeast, traffic from the trans-Golgi network to the surface is regulated by Ypt32p and Sec4p2, homologs of mammalian Rab11 and Rab8, respectively. These are linked by Sec2p (homolog of mammalian Rabin8), a guanine nucleotide exchange factor (GEF) for Sec4p, which is recruited by Ypt32p. Sec2p and Sec4p in turn interact with the exocyst, which docks vesicles to the surface3.
Definition of cortical domains in metazoa involves a complex of Par3, Par6, atypical PKC (aPKC), and the GTPase Cdc424. This complex is a master regulator of polarity, conventionally depicted upstream of membrane trafficking machinery. How this complex interfaces with membrane transport is poorly understood.
Here we show a molecular mechanism for lumen and apical surface formation, linking Rab8a/11a, exocyst, annexin2, Cdc42 and its GEF Tuba, and the Par3/aPKC complex. This novel pathway shows how the membrane traffic and cortical polarity machineries cooperate to generate the apical surface and lumen.
Upon plating into 3D culture, individual MDCK cells proliferate and assemble into cyst structures - a polarized spherical monolayer surrounding a central lumen. Lumenogenesis requires the apical membrane determinant gp135/podocalyxin5 (PCX in figures). Initially, MDCK aggregates have podocalyxin at the ECM-contacting surface (Fig. 1a, 12 h; Fig. S1a), before polarity inversion occurs, with β-catenin and Na/K-ATPase at cell-cell junctions and podocalyxin now at the lumen (Fig. 1a, 24–48 h, arrows; Fig. S1d)6, 7. Early lumens occur at a site previously termed the "Pre-Apical Patch" (PAP), where opposing plasma membranes are separated, but the podocalyxin signal is non-resolvable by confocal microscopy7; expansion enables luminal space visualization (Fig. 1b). In contrast, apical proteins syntaxin-3 and GFP-CNT1 (Concentrative Nucleoside Transporter-1) label the entire surface before concentrating at the lumen (Fig. S1b–c, e–f)5.
Binding of antibodies to GFP-VSVG-podocalyxin at the periphery of cysts, then incubating further to allow lumenogenesis to occur, revealed that podocalyxin at the PAP (arrowheads) and in vesicles (arrows) is at least partially derived from transcytosed peripheral podocalyxin (Fig. S1g).
Transcytosis of podocalyxin to the surface to establish the PAP represents formation of apical-basal polarization. We thus examined localization of select polarity (Par3/aPKC), trafficking (exocyst complex; Sec8/Sec10/Sec15A), and junctional (occludin) proteins during lumen initiation. Strikingly, although these proteins showed differing localizations before lumen formation, all converged transiently during lumen initiation (Fig. 1c–k; Table 1). When GFP-podocalyxin was peripheral, Par3 and Sec8 colocalized in puncta at the edge of cell-cell contacts (Fig. 1c, arrowheads). When GFP-podocalyxin was internalized and transcytosed some Par3 and Sec8 concentrated at the first detectable site of GFP-podocalyxin delivery to the nascent apical surface (Fig. 1d, arrow). We term this the Apical Membrane Initiation Site (AMIS). Later, Par3 and Sec8 instead enriched at the tight junction (Fig. 1e, arrowheads). We define early apical structures where several tight junction markers have become distinctly localized from podocalyxin as the PAP (see below)7, rather than the earlier AMIS, where tight junction markers are not resolved from podocalyxin by confocal.
Sec10 and occludin, in contrast, initially localized along the entire cell-cell contact in early aggregates with peripheral podocalyxin (Fig. S2a; data not shown). Podocalyxin delivered to the AMIS partially overlapped with Sec10 and occludin (Fig. 1f; Fig. S2b; arrows). Though occludin remained along the entire contact, Sec10 condensed toward the AMIS. As the lumen expanded, Sec10 and occludin enriched at the tight junction (Fig. 1g–h; Fig. S2c–f), though some occludin remained along cell-cell contacts (Fig. 1h).
aPKC follows yet a different pattern, with distinct pools initially localized with peripheral podocalyxin (arrowheads) and the AMIS (Fig. 1i, arrows), before enriching at PAP edges and finally the tight junction and lumen (Fig. 1j–k, arrowheads). Together, these data show the complex movement of trafficking and cortical polarity proteins, converging transiently at the AMIS.
We examined potential control of AMIS and lumen formation upon perturbation of select Rab GTPases involved in apical, basolateral, or junctional trafficking (Fig. 2a–f; Fig. S3f–l). In contrast to control cysts with a single lumen, apical podocalyxin and basolateral β-catenin, knockdown or dominant negatives of Rab8 (Rab8a/b) and Rab11 (Rab11a/25, but not Rab11b) family members significantly decreased single lumenogenesis (Fig 2f), instead displaying multi-lumens and accumulating podocalyxin in vesicles (Fig. 2b–e, arrowheads; data not shown) close to the cell surface (marked by β-catenin). For single lumen-perturbing knockdowns, phenotypes were confirmed using additional shRNAs (Fig. S3l), and additional cargos (Fig. S3a–c). Rab10, Rab11b, Rab13, and Rab14 perturbation did not overtly perturb lumenogenesis and were not pursued (Fig. 2f).
The Rab11 family regulates transcytosis8 and lumenogenesis in diverse systems9–11. GFP-Rab11a localized to vesicles underlying the AMIS (marked by Par3; arrow in Fig. 2g), then remained on subapical vesicles once lumens expanded (Fig. 2h; Table 1). Podocalyxin transcytosed to the AMIS via Rab11a vesicles. When podocalyxin was peripheral (arrow), GFP-Rab11a localized to juxtanuclear and peripheral vesicles (Fig. 2i, white and yellow arrowheads, respectively). Upon internalization, podocalyxin localized to GFP-Rab11a vesicles (Fig. 2j, arrowheads), then both were delivered to the cyst interior (Fig. 2k, arrowheads). Here, regions of podocalyxin devoid of GFP-Rab11a began to emerge (Fig. 2k–m), representing podocalyxin surface delivery (arrows). Similarly, IgA transcytosed to the PAP (Fig. S1h). As the lumen expanded, GFP-Rab11a clustered underneath the apical surface (Fig. 2l–m). Notably, overexpression of GFP-Rab11a (WT or activated Q70L) increased single lumenogenesis, while dominant negative GFP-Rab11aS25N attenuated single lumens and accumulated podocalyxin intracellularly (Fig. S4a; data not shown). Thus, Rab11a promotes transcytosis to the AMIS and single lumenogenesis.
Rab8 family GTPases were also required for single lumen formation (Fig. 2b–c, f), and Rab8a localized to transcytosing podocalyxin vesicles and the AMIS7 (Table 1; data not shown). Knockdown of Rab11a caused upregulation of Rab8a, and vice versa (Fig. S3f–g), suggesting compensation or co-operation between Rab11 and Rab8 families. To this end, we knocked down Rab8 and Rab11 family members, alone or in combination (Fig. S4b). Of tested combinations, co-knockdown of Rab8a/b, with or without Rab11a knockdown, most severely reduced single lumenogenesis. This suggests that the Rab8 family may act downstream of Rab11a. Accordingly, Rab8a knockdown blocked GFP-Rab11aQ70L-induced increased single lumenogenesis (Fig. S4c). These data are consistent with hypothesis that the Rab8 family acts, at least in part, downstream of Rab11a during AP transport and lumenogenesis, though the precise interaction between these Rabs may be more complex.
Rab11 binds the Rab GEF Rabin8 and stimulates its activity towards Rab812. We reasoned that Rab11a may control subapical Rabin8/Rab8 targeting. In control cysts, a small pool of Rabin8, and to a lesser extent Rab8a localized to dispersed puncta, with some clustered subapically (Fig. 3a, arrows). Expression of GFP-Rab11aWT, but not GFP-Rab11aS25N, strongly enhanced recruitment of Rabin8 and Rab8a to Rab11a-positive subapical vesicles (Fig. 3a, arrowheads; see also Fig. 5k for colocalization). Like endogenous Rab8a (Fig. 3a), GFP-Rab8aWT was cytoplasmic and in subapical vesicles, the latter of which was enhanced upon activated GFP-Rab8aQ67L expression (Fig. S4f). Thus, active Rab11a recruits active Rab8a to subapical vesicles, likely through Rabin8.
MDCK Rabin8 appeared as two bands, corresponding to its α and β isoforms: both possess the Rab11-binding region12 (Fig. 3b; Fig. S5a). Rabin8α knockdown accumulated some podocalyxin in vesicles (Fig. 3b–c, arrowheads), and although only to a modest level, significantly decreased single lumenogenesis (Fig. 3e). This modest effect is likely due to compensatory up-regulation of the Rabin8β isoform observed upon Rabin8α knockdown (Fig. 3b). Dual α/β knockdown caused severe cell death, precluding further analysis (not shown). Expression of RNAi-resistant GFP-hRabin8αWT, which localized to the luminal region (arrows), in endogenous Rabin8α knockdown cysts restored single lumenogenesis and podocalyxin localization (Fig. 3d–e). In contrast Rabin8α GEF domain mutants (Fig. S5a–c) further decreased single lumenogenesis and co-accumulated with podocalyxin on vesicles (Fig. 3d–e, arrowheads) beneath the surface marked by F-actin. Similarly, overexpression of GFP-TBC1D30WT, a GAP specific to the Rab8 family13, but not GAP-deficient GFP-TBC1D30R140A, perturbed single lumenogenesis (Fig. S5d–e). These data suggest that a Rab11a-Rabin8α-Rab8a cascade, experimentally opposed by TBC1D30, is part of a regulatory module governing apical transport and lumenogenesis.
Rab8a and Rab11a associate with the Sec15 exocyst subunit14, in turn linking to Sec10 and other subunits as part of a chain tethering vesicles to the basolateral3 and apical membranes15. The exocyst also interacts with the Par3/aPKC complex16, 17. As these factors converge at the AMIS, we examined their requirement in apical traffic and single lumenogenesis.
In contrast to control cysts with apical podocalyxin and basolateral β-catenin (Fig. 4a), Sec15A knockdown (Fig. S3m) accumulated podocalyxin in prominent, GFP-Rab11a positive vesicles close to the surface (Figs. 4b, arrows; 4c arrowheads); this Rab11a compartment seemed expanded relative to control cysts (compare Fig. 2m). Additionally, cysts were defective in apical polarization, mistargeting apical cargo to regions of cell-cell contact (Fig. 4b; Fig. S3d). Accordingly, Sec15A knockdown caused almost complete loss of single lumenogenesis (Fig. 4d). Similarly, Sec10 knockdown decreased single lumenogenesis and caused vesicular accumulation of podocalyxin (Fig. S2g–i). Thus, the exocyst regulates podocalyxin transport from Rab11a vesicles to the forming apical surface.
We also examined the role of the exocyst on Par3 transport and AMIS formation. In control cysts, Par3 localized to tight junctions (Fig. 4e, arrows). In Sec15A knockdown cysts, Par3 showed varying, though always abnormal, localization. In regions where a PAP formed, Par3 was recruited to the surface (Fig. 4f). However, in regions of vesicular podocalyxin accumulation, Par3 failed to be recruited to the surface and an AMIS was undetectable (Fig. 4f, arrowhead). Expression of RNAi-resistant GFP-Sec15AWT, which localized to subapical vesicles (Fig. 4g), in cysts with endogenous Sec15A knockdown rescued single lumenogenesis, and surface delivery of podocalyxin and Par3 localization (i.e. at tight junctions once lumens had formed).
To test the role of exocyst coupling to Rabs, we used GFP-Sec15AN691A, a Rab11-uncoupled mutant15. This mutant was completely unable to rescue the trafficking and single lumenogenesis defects caused by knockdown of endogenous Sec15A (Fig. 4h). Thus, coupling of exocyst to Rab8/11 is required for surface targeting of podocalyxin and Par3 to the AMIS.
Similar to exocyst knockdown, Par3 knockdown also resulted in intracellular podocalyxin accumulation close to the surface marked by β-catenin (Fig. 4i, arrows), in vesicles co-labeled for GFP-Rab11a (Fig. 4j, arrows), and a strong disruption of single lumenogenesis (Fig 4l). Par3 knockdown also mistargeted some GFP-CNT1 to cell-cell contacts (Fig. S3e). Moreover, upon Par3 knockdown, Sec8 was not recruited to surface regions adjacent to vesicular podocalyxin (Fig. 4k), representing a failure to form the AMIS.
Inhibition of aPKC, using its pseudosubstrate inhibitor (aPKC-PS; Fig 4m–n) similarly perturbed AMIS and single lumenogenesis, causing accumulation of podocalyxin in Rab11a-positive vesicles close to the surface marked by β-catenin (Fig. 4n, arrowheads). In addition, aPKC inhibition caused lack of podocalyxin internalization from the periphery in some cells (Fig. 4n, arrows), likely representing an additional function of aPKC at this locale (see Fig. 1i). Together, these data demonstrate a crucial role for the exocyst/Par3/aPKC complex in podocalyxin delivery from Rab11a vesicles to form the lumen.
Luminal targeting of aPKC in MDCK cysts requires interaction of GTP-Cdc42 with the PI(4,5)P2-binding protein Annexin2 (Anx2)6. Anx2 both transits to the surface via, and regulates the function of, Rab11a recycling vesicles18, 19. We thus examined interplay between Anx2, Cdc42 and the Rab11a-Rab8a module.
In early cysts with peripheral podocalyxin (arrow), and subperipheral Apple-Rab11a, GFP-Anx2 localized to the surface (Fig. S6a). When podocalyxin was in condensed Rab11a vesicles beneath the AMIS, some GFP-Anx2 now also localized to these vesicles (Fig. S6b, arrowheads). Once podocalyxin was at the open lumen (Fig. S6c, arrow), GFP-Anx2 localized to both apical and basolateral surfaces, but no longer to subapical Rab11a vesicles. Thus, GFP-Anx2 transiently associates with Rab11a vesicles during lumen initiation.
In contrast to controls expressing Anx2 WT with luminal podocalyxin, subapical Apple-Rab11a, and apical and basolateral GFP-Anx2 (Fig. S6d, f), expression of dominant negative Anx2 (Anx2 XM) perturbed lumenogenesis and caused the accumulation of GFP-Anx2 with podocalyxin in Rab11a-positive vesicles (Fig. S6e, arrowheads). Conversely, knockdown of Rab8a or Rab11a caused intracellular accumulation of podocalyxin in structures co-labeled with GFP-Anx2 (Fig. S6g–h, arrowheads). Thus, Anx2 and Rab8a/11a co-operate in the delivery of podocalyxin to the surface.
We next examined whether Cdc42 associated with Rab11a vesicles. Unlike Anx2, GFP-Cdc42, though possessing a large cytoplasmic pool, strongly overlapped with subapical Apple-Rab11a in cysts with open lumens (Fig. 5a, arrowheads). Activated Cdc42 (GFP-Cdc42Q61L) localized to cell-cell contacts and the luminal region, marked by podocalyxin (Fig. 5b; arrowheads). As GFP-Cdc42Q61L removed cytoplasmic background labeling, and expression did not perturb single lumens (Fig. 6i), we used this allele to further examine Cdc42 localization. In early cysts with peripheral podocalyxin (arrows), GFP-Cdc42Q61L localized to the surface (Fig. 5c). When podocalyxin was internalized into Rab11a vesicles and subsequently concentrated at the AMIS, GFP-Cdc42Q61L now extensively overlapped with these vesicles (Fig. 5d–e, arrowheads). As the PAP (Fig. 5f) and open lumen (Fig. 5g) formed, podocalyxin and Rab11a largely no longer overlapped, whilst GFP-Cdc42Q61L maintained some overlap with both (arrows). Thus, active Cdc42 associates with Rab11a vesicles during lumenogenesis.
We next determined whether Cdc42 is required for transport from Rab11a vesicles. As shown previously, Cdc42 knockdown perturbed lumenogenesis (Fig. 5n), causing accumulation of podocalyxin in VACS (arrows) or vesicles (arrowheads) close to the surface marked by β-catenin (Fig. 5i)6. Notably, intracellular podocalyxin observed upon Cdc42 knockdown was localized to Rab8a/11a vesicles, suggesting Cdc42 regulates transport from these vesicles (Fig. 5l, arrowheads).
We20 and Qin et al21 identified Intersectin-2 and Tuba as the only Cdc42-specific GEFs essential for MDCK lumenogenesis. As Intersectin-2 knockdown did not disrupt transport of podocalyxin in cysts20, we examined if Tuba regulates Cdc42-dependent podocalyxin transport. Tuba knockdown phenocopied Cdc42 knockdown, disrupted single lumenogenesis, and accumulated podocalyxin in Rab8a/11a vesicles (Fig. 5j,m–n, arrowheads). Notably, Tuba, and to greater extent Cdc42, knockdown blocked GFP-Rab11a-induced increased single lumenogenesis (Fig. 5n), suggesting that Rab11a operates upstream of both Tuba and Cdc42. Thus, Tuba-dependent Cdc42 activation is required for podocalyxin apical transport.
Tuba is required for Cdc42 apical targeting21. We examined whether Rab8a/11a also influenced Cdc42 activation. Rab8a, but not Rab11a, knockdown strikingly decreased global GTP-Cdc42 levels (Fig. 6a). Similarly, overexpression of GFP-Rab8aQ67L, but not GFP-Rab11aQ70L, robustly activated Cdc42 (Fig. 6b), suggesting that Rab8a influences global Cdc42 activation.
We examined whether Rab8a/Rab11a regulate apical Cdc42 targeting. A PBD-YFP probe of activated Cdc426 labeled the luminal surface, along with podocalyxin (Fig. 6c, arrowheads), and to a lesser extent cell-cell contacts, mirroring activated Cdc42 localization (Fig. 5b). Rab8a knockdown abrogated PBD-YFP membrane association, despite retaining luminal podocalyxin labeling (Fig. 6d, arrowhead). Strikingly, Rab11a knockdown resulted in a loss of apical (arrowhead), but not basolateral, PBD-YFP (arrows) (Fig. 6c). Thus, Rab8a is required for global activation and surface targeting of Cdc42, while Rab11a controls apical Rab8a, and consequently, active Cdc42 targeting.
We reasoned that as Rab8a/11a influenced apical targeting of active Cdc42, overexpression of active Cdc42 may rescue single lumenogenesis upon Rab8a/11a knockdown. Indeed expression of active Cdc42 (GFP-Cdc42Q61L) rescued apical targeting of podocalyxin (Fig. 6g–h) and single lumenogenesis in cysts with Rab8a or Rab11a knockdown (Fig. 6i). These data support the conclusion that Cdc42, regulated by Rab8a/11a, is required for apical transport of podocalyxin. Taken together, Rab11a regulates a molecular network directing the apical polarity and trafficking machineries to initiate de novo lumen formation.
How membrane trafficking and polarity complex machineries work together to form the apical surface and lumen is a fundamental issue22. We describe a molecular chain linking membrane trafficking machinery with delivery of the Par3/aPKC/Cdc42 complex to the nascent apical surface, from early cell aggregates, to apical membrane initiation (AMIS), formation of a PAP, then to cysts with open lumens (Fig. 7a). This emphasizes the complex spatiotemporal orchestration needed to construct a new membrane (see Table 1).
Podocalyxin is initially at the ECM-contacting periphery (Fig. 7a), before internalization into Rab11a vesicles, to which Rab8a is recruited via the GEF Rabin8 (and opposed by the GAP, TBC1D30) (Fig. 7b). Apical vesicle delivery and lumenogenesis is regulated by both Rab proteins. The exocyst, a Rab effector, docks vesicles with the surface to create the AMIS, in cooperation with Par3/aPKC (Fig. 7b). Anx2 and Cdc42 associate with Rab8a/11a vesicles, regulating apical transport and single lumenogenesis, dependent on the Cdc42 GEF Tuba. Par6 likely bridges Cdc42 to the aPKC/Par3/exocyst complex at the AMIS. Thus, apical polarity and membrane domain identity is initiated de novo by membrane delivery from Rab8a/11a vesicles. Finally, the luminal space is expanded by pumps and channels (Fig. 7k)7, 23.
Podocalyxin is the earliest marker of apical polarization that we have studied, and its initial appearance at the nascent apical surface marks the AMIS. Notably, the Rab11a-Rabin8α-Rab8a cascade also regulates mammalian ciliogenesis, and the homologous yeast pathway regulates budding24, suggesting it is an ancient polarity-generating module.
The exocyst is a Rab effector for transport of podocalyxin from Rab11 endosomes. This transport also required the Par3/aPKC polarity complex, revealing a mutual interdependence of these complexes for localization to the AMIS. Similarly, in developing neurites, the exocyst and Par3/aPKC associate17 with the exocyst required for Par3 localization to growing neurite tips. Interaction of the exocyst with Par3/aPKC may focus exocyst-dependent vesicle docking events transiently to the AMIS, allowing initiation of apical polarity, before both complexes relocalize to the tight junction for subsequent transport events. That the exocyst was also required for Par3 localization demonstrates that vesicle trafficking operates both upstream and downstream of cortical polarity proteins, suggesting a feedback loop.
Rab11 vesicles also deliver the Crumbs3-Pals1-PatJ complex to early lumens25. In Drosophila Crumbs acts to dissociate Par3 from Par6 and aPKC at the apical surface, restricting Par3 to apicolateral borders26, 27. Similarly, Crumbs3 delivered to the AMIS may exclude a pool of Par3 from the nascent apical surface, allowing it to concentrate to the sides of the developing lumens, such as in the transition from AMIS to PAP reported here.
aPKC/Par6 form a complex with Cdc4228. Cdc42, in conjunction with Anx26, regulates vesicular transport to the apical surface from Rab8a/11a vesicles. Cdc42, however, also regulates the orientation of cell division in conjunction with Par6/aPKC20, 21, 29. Indeed, the Cdc42 GEFs Intersectin-2 and Tuba regulate Cdc42 activation and division orientation during MDCK lumenogenesis20, 21. Intersectin-2 knockdown causes multi-lumens without vesicular podocalyxin accumulation20, suggesting it functions in cell division but not vesicle transport. In contrast, Tuba regulates apical targeting of Cdc4221 and, as demonstrated here, podocalyxin transport from Rab8a/11a vesicles.
Active Cdc42 localized to Rab11a vesicles. In yeast, Cdc42 is also delivered to the bud site30, suggesting that vesicular transport of Cdc42 to membrane being generated de novo is a conserved polarity-generating event. Notably, Rab8a/11a were required for Cdc42 activation at the lumen. Rab8a regulated global Cdc42 activation; Rab11a regulated apical Cdc42 targeting. That global Cdc42 activation was greatly decreased upon Rab8a but not Rab11a knockdown, suggests that while Tuba acts downstream of Rab11 and regulates a pool of Cdc42 activation on Rab11a vesicles, Rab8a may influence additional GEF proteins, such as Intersectin-2. How Rab8a/11a influence Cdc42 GEFs remains to be elucidated. Similarly, we demonstrate a role for aPKC in apical transport from Rab11 vesicles, suggesting that the Cdc42-Par6-aPKC complex may function, in addition to in cell division, in membrane transport31, 32. These data reveal a novel role for Anx2-Cdc42-aPKC-Par3 in conjunction with the exocyst and Tuba, in apical transport to the AMIS.
Our perturbations yielded multi-lumens and accumulation of apical proteins in subapical vesicles. Perturbation of apical traffic could cause multiple lumen formation via several overlapping mechanisms. Reduced apical delivery may prevent initially small lumens from enlarging and consolidating into one central lumen, which occurs in several mammalian organs33. Multiple small lumens also require ion pumping to hydrostatically enlarge and coalesce lumens. Apical trafficking defects could cause defective junctions and/or mislocalization of pumps and channels, preventing enlargement23.
Orientation of mitosis also regulates lumen formation. In Caco-2 cysts, Cdc42 knockdown correlates with disrupted spindle orientation, without notable apical transport defects29. Caco-2 lumen formation requires artificially increasing cAMP, which strongly promotes apical exocytosis34, potentially masking detection of apical transport defects. As Rab8/11, exocyst, and Par3/aPKC also have roles in spindle orientation and/or cytokinesis35–37, the extent to which their function in cell division is separate from apical trafficking is unknown. Apical membrane traffic might be needed to localize proteins that orient mitosis, such as Cdc42, aPKC and LGN20, 21, 38, 39. Notably, multi-lumens can occur without disruption to apical transport20, 39, suggesting that the two processes can be uncoupled. For example, in MDCK and other systems, lumens can form in the absence of cell division or apoptosis7, 40–43, instead requiring vesicular transport of podocalyxin to the cyst interior. Thus, coordination of division orientation and apical transport mechanisms are likely to be key to generating a single lumen.
Recently, the Par proteins, as well as aPKC and Cdc42, have been demonstrated as regulators of polarity, via endocytosis32. We demonstrate that the membrane traffic, especially exocytosis, is both upstream and downstream of the Par complex. Our data buttress an emerging view of the Par complex as a multifunctional platform modulating membrane traffic31, 32, and suggest Cdc42/aPKC/Par3 as a convergence between the machineries of cortical polarization and vesicular transport.
MDCK cysts were subcultured in 5% FBS (Gibco, Carlsbad, CA) MEM or grown in 3D Matrigel cultures (BD, San Jose, CA), as described6. Cells were trypsinized to a single cell suspension at 1.5 × 104 cells/ml in complete medium containing 2% Matrigel. Suspensions (250 μl) were plated into 8-well coverglass chambers (Nunc, Rochester, NY), pre-coated with 5 μl of 100% Matrigel. Cells were grown for 24–48 h before fixation in 4% PFA. In some experiments, cells were treated with aPKC-PS (40 μM; Invitrogen, Carlsbad, CA) from the time of plating to inhibit aPKCs6. In most instances, exogenous proteins were from stably expressing cell lines. For Anx2 XM experiments, cells in 2D were transiently transfected with Anx2 XM (Lipofectamine 2000, Invitrogen, Carlsbad, CA) 24 h prior to plating into 3D.
Stable RNAi was achieved by viral shRNA. For Sec10 knockdown, cells were transfected with siRNA oligonucleotides as described6. In all instances knockdown was verified by standard western blot or Q-RT PCR procedures (Brilliant-II SYBR Green Kit, Agilent, Santa Clara, CA), normalizing to GAPDH expression. Q-RT PCR Primers are presented in Supplementary Table 1. RNAi target sequences are presented in Supplementary Table 2. Rab8a_1, Rab10, Rab11a_1, Rab11b, in pRVH1-puro or –hygro retroviruses were previously published44. All other shRNAs were generated in pLKO.1-puro45, or pLKO.1-blast, which was constructed by exchanging the puromycin resistance gene for blasticidin. Cdc42, Par3 and Tuba shRNAs were adapted for pLKO.1 from published sequences21, 46. pLKO.1 lentiviruses were constructed according to the Addgene pLKO.1 protocol (www.addgene.org) using iRNAi (www.mekentosj.com), and target sequences were based on an (AA)N19 algorithm. RNAi sequences were submitted to BLAST (NCBI) to verify target specificity. For isoform-specific RNAi to Rabin8, shRNAs predicted to target the α isoform (Rabin8_4, Rabin8_5) or to both α and β (Rabin8_2) isoforms of canine Rabin8 were extrapolated from sequence alignment with human Rabin8 splice forms (mined from NCBI). For knockdown and rescue experiments, GFP-tagged plasmids of transcripts from human or rat, which are not targeted by anti-canine shRNAs, were used.
Retrovirus production was performed essentially as previously described 44, except pRVH1 plasmids were transfected into 293-GPG cells (O. Weiner, UCSF)47. 48 h post transfection, viral supernatants were collected daily for 7 d. For lentivirus production, pLKO.1 plasmids were co-transfected with ViraPower packaging mix into 293-FT cells according to manufacturer's protocols (Invitrogen, Carlsbad, CA). All supernatants were centrifuged to remove cell debris and frozen in liquid nitrogen for further use.
For retrovirus transduction (pRVH1), subconfluent cultures of MDCK cells, 16 h after plating, were incubated with virus-containing supernatants supplemented with 10 μg/ml Polybrene (Millipore, Billerica, MA) for 24 h at 32°C. Upon changing to fresh medium, cells were incubated for a further 24 h at 37°C, before passage into appropriate antibiotic-containing medium. For lentivirus transduction (pLKO.1), subconfluent MDCK cultures, 1–4 h after plating, were infected with virus-containing supernatants for 12–16 h at 37°C. Viral supernatants were then diluted 1:1 with growth medium, cultured for a further 48 h, then passaged into appropriate antibiotic-containing medium. Hygromycin (0.5 mg/ml), puromycin (5 μg/ml), and blasticidin (12.5 μg/ml) were used.
Plasmids kindly provided were: GFP-Cdc42 (Q61L; Addgene, Cambridge, MA), GFP-Rab11a (WT, S22N; E. Brown, UCSF), GFP-Rab8a (WT, Q67L), GFP-Sec15A (J. Stow, U. Queensland, Australia), GFP-VSVG-podocalyxin, pRVH1-puro/hygro (control, Rab8a, Rab10, Rab11a and Rab11b; K. Simons, EMBL), GFP-TBC1D30 (WT, R140A; F. Barr, U. Liverpool). GFP-Cdc42, YFP-PBD, GFP-Annexin2 (WT, XM)6, GFP-Rab13 (T22N; A. Zharaoui, CEA CNRS, France), GFP-Rab14 (S25N; J. Wilson, U. Arizona), GFP-Rabin8α48, Syntaxin3-2xmyc (T Weimbs, UCSD)49, GFP-CNT150 cell lines/plasmids as described. All additional plasmids were constructed through site-directed mutagenesis (Quikchange, Agilent, Santa Clara, CA) or standard subcloning. For generation of stable lines, transfected cells underwent FACS after selection to obtain appropriate expression levels.
Primary mouse antibodies used were: Cdc42 (BD Biosciences, San Jose, CA) [Dilution: Western Blot (WB), 1:1000]; GAPDH (Millipore, Billerica, MA) [Dilution: WB, 1:10,000]; gp135/podocalyxin (G. Ojakian) [Dilution: IF, 1: 1000]; myc (9E10, Santa Cruz Biotechnology, Santa Cruz, CA) [Dilution: IF, 1:200]; p58/Na/K-ATPase (K. Matlin) [Dilution: IF, 1:200]; Tuba (Abnova, Taipei City, Taiwan) [Dilution: WB, 1:500]; VSVG (P5D4) [Dilution: Transcytosis assay, 1:1000] 51. Primary rabbit antibodies used were: aPKCζ (C-20) [Dilution: IF, 1:200]; β-catenin (Santa Cruz Biotechnology, Santa Cruz, CA) [Dilution: IF, 1:200]; GFP (Invitrogen, Carlsbad, CA) [Dilution: Transcytosis assay, 1:1000]; Occludin (Invitrogen, Carlsbad, CA) [Dilution: IF, 1:200]; Par3 (Millipore, Billerica, MA) [Dilution: IF, 1:100; WB, 1:1000]; Rabin8 [Dilution: IF, 1:50; WB, 1:500]; and Rab8a [Dilution: IF, 1:50; WB, 1:500]; 48; Rab8b (ProteinTech, Chicago, IL) [Dilution: WB, 1:500]; Rab10 (Sigma, St. Louis, MO) [Dilution: WB, 1:500]; Rab11a (Millipore, Billerica, MA) [Dilution: IF, 1:100; WB, 1:1000]; Rab25 (Cell Signaling Technology, Danvers, MA) [Dilution: WB, 1:500]; Sec8 (Enzo Life Sciences, Plymouth Meeting, PA) [Dilution: IF, 1:100]; Sec10 [Dilution: IF, 1:250 (fix in MeOH:acetic acid; WB, 1:250] (W. Guo). Rat anti-ZO-1 (R40.76; B. Stevenson) [Dilution: IF, 1:200]; was also used. Alexa Fluorophore-conjugated secondary antibodies [Dilution: IF, 1:250 for all secondary antibodies] or Phalloidin (Invitrogen, Carlsbad, CA) [Dilution: IF, 1:400], and Hoescht to label nuclei [Dilution: 10 μg/ml], were utilized. Cyst were stained essentially as previously described6.
Single lumen formation was quantified as described6. The percentage of cysts with a single lumen was determined, and normalized to control cysts as 1.0. Values are mean ± SD from 3 replicate experiments, with n ≥100 cysts/replicate. Significance was calculated using a paired, two-tailed Student's t-test. *p < 0.05, ** < 0.001, *** < 0.0001.
For PCX transcytosis, cells stably expressing GFP-VSVG-podocalyxin were plated into 8-well chamber slides and grown 12 h at 37°C. Slides were placed on ice and washed twice with serum-free medium at 4°C before incubation at 4°C for 30 min with serum-free medium containing indicated antibodies. Cysts were then washed twice with cold serum-free medium, before being grown for 24 h at 37°C in fresh serum-containing medium supplemented with 2% Matrigel.
pIgA, kindly provided by Professor J.P. Vaerman (Catholic University of Louvain, Brussels, Belgium), was biotinylated using sulfo-NHS-LC-biotin (Pierce, Rockford, IL). For IgA uptake, MDCK PTR-9 cells stably expressing hTfR and rabbit pIgR52 were grown for 36 h in 3D to induce cyst formation, then incubated with 100 μg/ml biotinylated IgA in complete medium for 40 min. Biotinylated IgA was detected using fluorescently labeled streptavidin (Invitrogen, Carlsbad, CA).
GTP loading of Cdc42 was by GST-PAK pull down (Cytoskeleton, Denver, CO), according to manufacturer's protocol, with modifications. Cells were lysed in Mg Lysis Buffer (Millipore, Billerica, MA) containing 1 mM PMSF and protease inhibitor cocktail, lysates were passed through a 27 gauge needle (BD, Franklin Lakes, NJ), and cleared of debris by centrifugation. A sample of lysate was taken for protein concentration determination (BCA, BioRad, Hercules, CA), and lysates were snap-frozen. 3 μg protein from appropriate condition was incubated with 20 μg of GST-PAK bead for 30 min at 4°C. Beads were collected by centrifugation and washed thrice in buffer before SDS-PAGE. Total and GTP-Cdc42 levels were detected by blotting using a Cdc42 antibody (BD Biosciences, San Jose, CA).
GEF mutants of Rabin8 were modeled on critical, conserved residues in the Rab GEF domain governing Sec2p-Sec4p interactions53. L196A and F201A mutants were generated from pEGFP-C1-Rabin8α and pGEX-2T-Rabin8α48 (Quikchange, Agilent, Santa Clara, CA). Decreased, direct association of Rab8a with Rabin8 mutants was verified by GST pulldown. Briefly, GST-Rabin8, GST-Rabin8-L196A, GST-Rabin8-F201A protein (15°C overnight; 200 μM IPTG) or GST protein expression (37°C for 3 h) was induced in bacteria. Lysates containing GST fusion proteins were incubated with glutathione-agarosebeads (Sigma, St. Louis, MO) at 4°C for 2 h, then washed thrice during30 min with binding-buffer (50 mM Tris, pH 7.5, 150 mM NaCl, 2 mM MgCl2,1% Triton X-100). Rab8A-T22N and Rab8A-Q67L were translated in vitro using a TNT Quick kit (Promega, Madison, WI) accordingto the manufacturer's instructions. The in vitro translation productswere incubated with GST proteins coupled to glutathione agarose beads in binding buffer (50 mM Tris, pH 7.5, 150 mM NaCl, 2 mM MgCl2,1% Triton X-100) on a wheel at 4°C for 1 h. The beads were washed four times with binding-buffer over 30 min. Bound material was eluted from the beads with Laemmli sample buffer and loaded onto a 12% SDS-polyacrylamide gel. 1/10 of the in vitro translation reactions was used asa control. Bands were visualized by autoradiographyof the driedgels. Densitometry revealed that the L196A and F201A mutants decreased association with Rab8a by 39% and 52%, respectively, confirming their reduction-of-function characteristics.
We thank F. Barr, E. Brown, J. Stow, W. Guo, I. Macara, K. Simons, J. Wilson, T. Weimbs, and A. Zahraoui for generous gifts of reagents and unpublished data, and the Mostov lab for kind assistance. Supported by a Susan G Komen Foundation Fellowship (DMB), a DOD Breast Cancer Concept Award (AD), and NIH R01DK074398, R01AI25144 and P01AI53194 (KM).
The authors declare that they have no competing financial interests
AUTHOR CONTRIBUTIONSD.M.B., A.D., A.R.F, F.M.B., and J.P. designed the experiments; D.M.B., A.D., A.R.F, and J.P. did the experimental work; D.M.B. and K.E.M. analysed the experiments; D.M.B. and K.E.M wrote the manuscript.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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Is that a teller in your pocket or are you just happy to see me? New mobile banking tech lets you chat face to face with a teller on your mobile device.» Read more
The big selling point for arbitration has been that reducing legal costs by blocking customer lawsuits would result in lower prices for consumers. But that hasn't been the case.» Read more
To ensure the nation's largest banks won't crumble in the next big economic downturn, the Fed put the nation's biggest banks through a tough stress test.» Read more
Of any checking account feature, customers rated debit cards as the most essential. A whopping 60 percent said they couldn't image living without a debit card.» Read more
A new survey shows that young military members are positioning themselves for long-term personal finance success.» Read more
A new report released by the Treasury's Office of Financial Research scores banks on the scale of risk they pose to the financial system, and Chase came in at No. 1 overall by a wide margin.» Read more
As Valentine's Day approaches, a new study looks at how saving and spending philosophies impact relationships.» Read more
An EMV card with a smart chip loaded with encryption technology is difficult for anyone to clone at any price, let alone your run-of-the-mill petty criminal.» Read more
Is your family playing a dangerous game by keeping a low savings account balance? If so, you're not alone.» Read more
The Federal Reserve is forming task forces to combat the two biggest problems in America's '70s-era payment system: slow speeds and slack security.» Read more

subset: commoncrawl


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Data from: Weak evidence for anticipatory parental effects in plants and animals
Date Published: June 20, 2013
DOI: http://dx.doi.org/10.5061/dryad.pd5ss
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Lately, I've been doing behest every chance I can get. I'll usually go afk between them and do chores or homework. I wonder if that is why there are so many people afk at the atheryte =P I'm getting anywhere from 2.5k-5k SP in a period of 15mins with my rank 22 Gladiator. Anyways, I don't think this is common knowledge yet but a lot of people are catching on which is great. WE SHOULD BE HELPING EACH OTHER FIGURE THIS GAME OUT AND LESS TIME COMPLAINING ABOUT HOW BROKE IT IS.
Lately, I've been hoping to get into a solid group to knock out leves. I hear 'linking' them gives a significant SP bonus. Nevertheless, I've yet to be able to since it seems people would rather solo their leves. I guess only shot I would have to get into something like this would be to organize one ahead of time on our LS website >.>
I'm curious just how significant the SP bonus is and if it is worth the time and effort. I've heard that if two people in the group are linking a leve, you'll get x2 the SP, if three people, x3, and so on. That seems pretty crazy. I feel if more people knew this then we would see more parties forming and the search function being used more often. This would be the great incentive we would need to encourage group play. Another question I had is whether all members experience this SP bonus or if it is only experienced by the individuals that had the guild leve.
If all this is news to you, please reply as such. I'm really curious as to how many people aren't taking advantage of these two prominent ways to gain skill ups. Mindless grinding groups I've been in lately pale in comparison with this new update, especially past rank 20, but yet some people still insist on doing them. As easy mode as it sounds, I really wish there was less ambiguity in the game and SE would just tell us what there design and intention was to help adventures skill up.


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Series: Geophysical Monograph Series Volume: 222
426 pages, illustrations
Over a half century of exploration of the Earth's space environment, it has become evident that the interaction between the ionosphere and the magnetosphere plays a dominant role in the evolution and dynamics of magnetospheric plasmas and fields. Interestingly, it was recently discovered that this same interaction is of fundamental importance at other planets and moons throughout the solar system. Based on papers presented at an interdisciplinary AGU Chapman Conference at Yosemite National Park in February 2014, Magnetosphere-Ionosphere Coupling in the Solar System provides an intellectual and visual journey through our exploration and discovery of the paradigm-changing role that the ionosphere plays in determining the filling and dynamics of Earth and planetary environments. The 2014 Chapman conference marks the 40th anniversary of the initial magnetosphere-ionosphere coupling conference at Yosemite in 1974, and thus gives a four decade perspective of the progress of space science research in understanding these fundamental coupling processes. Digital video links to an online archive containing both the 1974 and 2014 meetings are presented throughout this volume for use as an historical resource by the international heliophysics and planetary science communities.
Topics covered in this volume include:
- Ionosphere as a source of magnetospheric plasma
- Effects of the low energy ionospheric plasma on the stability and creation of the more energetic plasmas
- The unified global modeling of the ionosphere and magnetosphere at the Earth and other planets
- New knowledge of these coupled interactions for heliophysicists and planetary scientists, with a cross-disciplinary approach involving advanced measurement and modeling techniques
Magnetosphere-Ionosphere Coupling in the Solar System is a valuable resource for researchers in the fields of space and planetary science, atmospheric science, space physics, astronomy, and geophysics.
Part I: Introduction
1. Magnetosphere-Ionosphere Coupling—Past to Future
James L. Burch
Part II: The Earth's Ionosphere As a Source
2. Thermal and low-energy ion outflows in and through the polar cap: The polar wind and the low-energy component of the cleft ion fountain
Andrew W. Yau, William K. Peterson, and Takumi Abe
3. Low-energy Ion Outflow Observed by Cluster: Utilizing the Spacecraft Potential
Stein Haaland, M. Andre, A. Eriksson K. Li, H. Nilsson, L. Baddeley, C. Johnsen, L. Maes, B. Lybekk, and A. Pedersen
4. Advances in Understanding Ionospheric Convection at High Latitudes
Roderick A. Heelis
5. Energetic and Dynamic Coupling of the Magnetosphere-Ionosphere-Thermosphere System
Gang Lu
6. The Impact of O+ on Magnetotail Dynamics
Lynn Kistler
7. Thermal and Low-Energy Ion Outflows In and Through the Polar Cap: The Polar Wind and the Low-Energy Component of the Cleft Ion Fountain
Naritoshi Kitamura, K. Seki, Y. Nishimura, T. Abe, M. Yamada, S. Watanabe, A. Kumamoto, A. Shinbori, and A. W. Yau
8. Ionospheric and Solar Wind Contributions to Magnetospheric Ion Density and Temperature Throughout the Magnetotail
Michael Liemohn and Daniel T. Welling
Part III: The Effect of Low Energy Plasma on the Stability of Energetic Plasmas
9. How Whistler-Mode Waves and Thermal Plasma Density Control the Global Distribution of the Diffuse Aurora and the Dynamical Evolution of Radiation Belt Electrons
Richard M. Thorne, Jacob Bortnik, Wen Li, Lunjin Chen, Bbinbin Ni, and Qianli Ma
10. Plasma Wave Measurements from the Van Allen Probes
George B. Hospodarsky, W. S. Kurth, C. A. Kletzing, S. R. Bounds, O. Santolik, R.M. Thorne, W. Li, T. F. Averkamp, J. R. Wygant, and J. W. Bonnell
11. Ring Current Ions Measured by the RBSPICE Instrument on the Van Allen Probes Mission
Louis J. Lanzerotti and Andrew Gerrard
12. Global Modeling of Wave Generation Processes in the Inner Magnetosphere
Vania K. Jordanova
Part IV: Unified Global Modeling of Ionosphere and Magnetosphere at Earth
13. Modeling Magnetosphere-Ionosphere Coupling via Ion Outflow: Past, Present, and Future
Robert W. Schunk
14. Coupling the Generalized Polar Wind Model to Global Magnetohydrodynamics: Initial Results
Daniel Welling, A. R. Barakat, J. V. Eccles, R. W. Schunk, and C. R. Chappell
15. Coupling Ionospheric Outflow into Magnetospheric Models: Transverse Heating From Wave-Particle Interactions
Alex Glocer
16. Modeling of the Evolution of Storm-Enhanced Density (SED) Plume during the Oct. 24-25, 2011 Geomagnetic Storm
Shasha Zou and Aaron J. Ridley
17. Forty-Seven Years of the Rice Convection Model
Richard Wolf, R. W. Spiro, S. Sazykin, F. R. Toffoletto, and J. Yang
18. Magnetospheric Model Performance During Conjugate Aurora
William Longley, Patricia Reiff, Jone Reistad and Nikolai Østgaard
19. Day-to-Day Variability of the Quiet-Time Plasmasphere Caused by Thermospheric Winds
Jonathan Krall, J. D. Huba, D. P. Drob, G. Crowley, and R. E. Denton
Part V: The Coupling of the Ionosphere and Magnetosphere at Other Planets and Moons in the Solar System
20. Magnetosphere-Ionosphere Coupling at Planets and Satellites
Thomas Cravens
21. Plasma Measurements at Non-Magnetic Solar System Bodies
Andrew Coates
22. Plasma Wave Observations with Cassini at Saturn
George B. Hospodarsky, D. Menietti, D. Pisa, W. S. Kurth, D. A. Gurnett, A. M. Persoon, J. S. Leisner, and T. F. Averkamp
23. Titan's Interaction with Saturn's Magnetosphere
Joseph H. Westlake, T.E. Cravens, R.E. Johnson, S. Ledvina, J.G. Luhmann, D.G. Mitchell, M.S. Richard, I. Sillanpää, S. Simon, D. Snowden, J.H. Waite, Jr., and A. K. Woodson
Part VI: The Unified Modeling of the Ionosphere and Magnetosphere at Other Planets and Moons in the Solar System
24. Magnetosphere-Ionosphere Coupling at Jupiter and Saturn
Thomas W.Hill
25. Global MHD Modeling of the Coupled Magnetosphere-Ionosphere System at Saturn
Xianzhe Jia, Margaret G. Kivelson, and Tamas I. Gombosi
26. Simulation Studies of Magnetosphere and Ionosphere Coupling in Saturn's Magnetosphere
Raymond J. Walker and Keiichiro Fukazawa
27. Characterizing the Enceladus Torus by Its Contribution to Saturn's Magnetosphere
Yingdong Jia, H. Y. Wei, and C. T. Russell
Part VII: Future Directions for Magnetosphere-Ionosphere Coupling Research
28. Future Atmosphere-Ionosphere-Magnetosphere Coupling Study Requirements
Thomas E. Moore, K. Brenneman, C.R. Chappell, J.H. Clemmons, G.A. Collinson, C.M. Cully, E. Donovan, G. D. Earle, D. Gershman, R.A. Heelis, L.M. Kistler, M.L. Kepko, G.V. Khazanov, D.J. Knudsen, M. Lessard, E. MacDonald, M.J. Nicolls, C.J. Pollock, R.F. Pfaff, D.E. Rowland, E. Sanchez, R.W. Schunk, J.L. Semeter, R. J. Strangeway, and J.P. Thayer
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text: November-7 November-7 is back with a NEW ALBUM and an innovative therapy!! Formed in 2005, November-7 are known for their powerful combination of industrial metal, fat guitar riffs and melodic vocals. Between 2006 and 2010 they release several records and DVDs and in 2011 they launch their first full-length studio album, Season 3 (Daily Rock Records / Musikvertrieb) mixed by the legendary Stefan Glaumann (Rammstein, Within Temptation, Deathstars). The record receives tremendous feedback both from the international media and the fans. November-7 open for well-established artists such as Marilyn Manson, Within Temptation, Sabaton, Pradise Lost, Lacuna Coil and in 2012 they tour the UK, Germany, Belgium, Austria and the Netherlands. In 2013 the band goes back to the studio and records a new album, Awaraxid 7mg, titled after a drug name. Why this title? « Increasingly more people are not happy with their life – explains Annamaria, singer and lyricist of the band – The older we get, the worse it becomes and we end up considering this as a natural consequence of adulthood. So we thought that it'd be great if there were a drug that could help us bring back our childhood dreams ». But once those dreams are re-discovered, it's up to each individual to realize them. In any case, they will not be able to go back to their previous life anymore and forget their dreams again. The album Awaraxid 7mg is pure November-7: it embraces melodies and martial sounds, fusing catchy choruses, fat and obsessive guitars and industrial samples. Stefan Glaumann's mix delivers each sound with extreme power and accuracy. Release date is on April 25th 2014 (Daily Rock Records / Musikvertrieb) and several concerts in Switzerland and in Europe are already confirmed. More will follow. Users who follow November-7 Elias Rodarte Elias Rodarte Serge Oleynikov-hutt Serge Oleynikov-hutt Riffenstein Riff Enstein SiNist3r_Vamp Jim Jay 6 Jim Jay pine ridge Natalie Chandra Natalie Chandra Zurich, Switzerland jbht Switzerland Hakan ertaş Hakan ertaş My Playlist Collection 2 Maik Baptist Hannover, Germany ShanSenpai


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How to buy original and high quality square wave inverter?
Posted by Rajanya Ganguly on
If the home makers are frequently experiencing the irritating situations of the power cuts and you need an alternate electrical source then buying the best type of home inverters for your home is highly significant to everyone. When it comes to the inverter models for your home appliances, there are several types of the inverters available in the market. Several new types of inverters are creating the different wave forms in order to give different ways of power supply to your household items. From among them, square wave inverters are the best choice for the house owners.
Some of the old style and the solid state home inverters are producing the square waves to supply the enough amount of electricity to your home appliances. Whenever your refrigerator, mixer grinder, television or any other home appliance require the constant force of electricity, the need of the square wave home inverters are very much important to switch the direct current into alternate current in the instant manner. It will give the sufficient electrical supply as the normal grid supply for the proper running of your household items with the proper voltage.
When the house owners have decided to buy the home inverters which are producing the square wave forms for the standard power supply for the household appliances, luminous is one and only a right choice of brand currently available in the market. The square wave inverters from our luminous brand are usually manufactured with the latest microprocessors, most advanced range of the PCB programming and also the strong FSW transformers in order to make it more efficient to store the power for your potential use. Our inverters will consume only lesser power for their functions of converting the current and storing them in the battery for your future use during the power outages at home.


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No.5 Wildcats Downed by No.3 South Carolina, 15-5
No.5 Wildcats Downed by No.3 South Carolina, 15-5
Sept. 20, 2013
COLUMBIA, S.C. – The No.5 Kansas State Equestrian team opened the 2013-14 season with a loss on Friday, after dropping a 15-5 decision to No.3 South Carolina.
"Congratulations to USC on their win today," said head coach Casie Maxwell. "Their riders were extremely prepared and rode very well. I can say without reservation that our team went out and rode the hardest they ever have for the first meet of the season. Their confidence and aggression was outstanding and what we want to build on for the rest of the season. We had a few errors which kept us from a few of our points, so I expect to see those riders adjust their plan for tomorrow's competitions. We look forward to adjusting from our mistakes and having the opportunity to continue building momentum in two road meets tomorrow."
The Hunter Seat team was bested 8-2 by the Gamecocks.
Captain Rachel Webster picked up the lone point for her team in Equitation on the Flat, defeating Kelsey Hart, 82-77. Freshman Alexis Graves had a successful Wildcat debut, knocking off Emily Reynolds, 79-76, in Equitation Over Fences.
K-State's Western team fell to USC, 7-3.
In Horsemanship, the Wildcats were led by Chayna DeNicolo and Kelly Bovaird. DeNicolo gave K-State their first point after taking down Kelsey Urban, 76-73. Bovaird answered with a strong ride of her own, giving her team another point with her 77-76.5 victory over Sam Chiodo. Freshman Alyssa Lombardi scored an exhibition win by besting Bailey Mierzejewski, 75.5-73.5. Kara Guy earned the Wildcat's only point in Reining after just narrowly defeating Cody McMillian, 67-66.5.
The Wildcats will look to bounce back on Saturday as the team travels to Dover, Del., for a showdown with Delaware State.


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Pier Village
Pier Village is a Victorian-inspired mixed-use community located in Long Branch, New Jersey along the Atlantic Ocean. It opened in 2005.
The land that Pier Village now sits on was originally that of the Long Branch Pier and the Kid's World amusement park.
On June 8, 1987, at around 3 PM, a fire broke out underneath the boardwalk in an electrical and gas box at the end of the pier. The fire quickly spread throughout the pier despite efforts by the Long Branch Fire Department to save it. After the fire only the water attractions across the street from the pier remained. In the following weeks business had slowed down so the remaining portion of Kid's World closed down permanently at the end of the summer.
The owner, Pat Cicalese, planned on rebuilding the pier. However, his insurance did not cover the cost to rebuild what was destroyed and the town would not fund the project so the plans were abandoned. What was left of Kid's World remained abandoned for 17 years after the fire.
Remains of the pier were demolished in the late 1990s. The rest of Kid's World across the street met the wrecking ball in 2002.
Applied Development Company, a Hoboken, NJ-based developer led by brothers David Barry and Michael Barry (the sons of Joseph Barry), was designated the developer of the 16-acre (65,000 m2) site by the City of Long Branch and soon began construction of Pier Village, starting with the rebuilding of the boardwalk.
Pier Village consists of 536 rental residences sitting atop more than 100,000 square feet (9,300 m2) of retail space. A public grassy area called Festival Plaza is the site of regular events, including concerts, arts & crafts fairs, outdoor movies and holiday events. The first story of Pier Village is filled with shops, restaurants, lounges, a salon, and a gym, while the upper floors are filled with ocean-view Apartments. There is two-hour metered surface parking and two enclosed parking garages.
The tenants are mostly local businesses, most from Monmouth County, but there are national brands, such as Stewart's Root Beer, Rocky Mountain Chocolate Factory, Red Mango, and Gold's Gym.
There are three restaurants situated on the boardwalk facing the ocean: Avenue, Sirena and McLoones. Of these, only McLoone's Pier House is free-standing. McLoone's was the only building built from the ground up by the restaurateur and was open a year in advance of the rest of the commercial properties within the Village. Also located in Pier Village is The Wine Loft a high end Wine Bar. The developers David Barry and Michael Barry, along with partners, opened up a members-only beach club called Le Club. Designed by London-based architect David Collins, who also designed London's Nobu Berkeley Street restaurant and the famed Blue Bar, Le Club is a 25,000-square-foot (2,300 m2) resort featuring a modern tri-level building with a rooftop pool and sun deck offering views of the ocean and a private beach area with palm trees and cabanas. Travel & Leisure Magazine named it one of "20 Great American Beaches" in its July, 2007 issue [1], and New York Magazine profiled it in its annual Summer issue [2].
The primary criticism of Pier Village was that Eminent Domain was abused to secure the land for development. Many argued that the homes seized were not "blighted" as the developer claimed. Residents were removed from their homes so that new homes could be constructed, which falls outside of the legal bounds of Eminent Domain, which states land can only be seized for public use.
The New York Times reported: "Either way, the fight has become a public-relations disaster for the city as it tries to reclaim its glory days, when seven United States presidents vacationed here and New York's business barons traveled by stagecoach to fill the gambling halls. The 30 or so holdout homeowners have planted signs saying NOT For Sale in their sandy yards, and they have filled the windows of their pastel bungalows with posters declaring Eminent Domain Abuse, signed petitions, shouted at public meetings and written vitriolic letters to the local newspaper questioning the mayor's and the City Council's motives." [1]
In 2010, the state ruled that the downtown area was not blighted, and thus could not be seized by Eminent Domain to be redeveloped.
The Urban Land Institute (ULI) named Pier Village "Project of the Year" in 2006 [3].
New Jersey Governor's Tourism Award in 2009 [4]
New pier[edit]
In 2005, plans for a new pier were unveiled.
The new plans would include:
- A ferry terminal
- An area for fishing
- A 30,000 sq ft (2,800 m2). area for exhibitions that would include a restaurant
- A beach-side stage
- An amphitheater
- A 3,500 to 5,000 sq ft (460 m2). area offshore that will be used for oceanographic and marine science educational classes
Hotel addition[edit]
Part of the Phase II expansion is Bungalow, a 25,000-square-foot (2,300 m2), 24-room boutique hotel with views of the ocean that is scheduled to open in May, 2009. Bungalow is much more modern than the original Victorian-styled buildings and sits at the entrance of Pier Village at the corner of Ocean Blvd. and Laird Street.
Pier Village Phase III[edit]
Applied Development Company is seeking approvals from the City for a third and final phase at Pier Village that will consist of a 100-room hotel, 200 rental residences, 75 condominium homes, 20,000 square feet (1,900 m2) of retail space and a new oceanfront public park.
In popular culture[edit]
- In a March 2008 episode of the ABC hidden-camera show, Primetime: What Would You Do?, which has actors do abnormal things (such as get into verbal fights, rob stores, etc.), showed an actor, who was significantly overweight, sitting on a bench in Pier Village while three other actors verbally insulted her.[5]
Pier Village is being developed by David Barry and Michael Barry, principals of Applied Development Company. The brothers are also principals of Ironstate Development. Other notable projects from them include:
- The W Hoboken Hotel & Residences, a new 26-story hotel offering 225 rooms and 40 condominium residences on the Hudson Riverfront in Hoboken, NJ facing mid-town Manhattan. The hotel is scheduled to open February, 2009;
- Port Liberte a 1,290-unit master-planned community in Jersey City, NJ. 804 homes have been built to date;
- 50 Columbus, a 35-story building offering 400 rental units, 950-car parking garage and 25,000 square feet (2,300 m2) of retail space in Jersey City, NJ;
- W Marrakech Hotel & Residences: a future hotel in Morocco, North Africa. The hotel will offer 150 hotel rooms and 50 whole-ownership villa units. The property is scheduled to open in mid-2010. W Marrakech Hotel & Residences will be a core element of the Al Maaden Resort development and will overlook an 18-hole championship golf course.
- Harrison Commons, a New Urbanist Master Planned community featuring more than 2,600 residences and 80,000 square feet (7,400 m2) of retail space in Harrison, NJ adjacent to the PATH Station. Sitework is underway.
- ^ George, Jason (31 March 2004). "Testing the Boundary Lines of Eminent Domain; Long Branch Wants to Seize Old Homes to Make Room for New Ones". Retrieved 18 May 2017 – via NYTimes.com.
External links[edit]
- Pier Village
- The Apartments at Pier Village
- New Pier Article - Darkinthepark.com
- Fall 2006 Expansion
- Article about expansion
- Asbury Park Press: The Good Life
- NY Times article about Pier Village's impact on Long Branch
- NY Times article about Phase II expansion
- NY Times article
- NY Post article about expansion and Le Club Beach Club
- Ironstate Development Website
Coordinates: 40°18′07″N 73°58′43″W / 40.301818°N 73.978736°W


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First quality bouncy wig made with skin center part and long, layered waves. 25" long.
Color available: #10/18: A Mix of 10-medium golden brown and 18-light strawberry blonde
Available in a 23" head size with skin top.
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What's the Catch?
Owning individual bonds has its risks and rewards. However, buying a bond entails an unseen transaction cost, which may not always be clear to retail investors. This transaction cost exists because bonds are not typically sold with a commission. Instead, a markup is built into the bond price.
This report offers a transparent look at these hidden transaction costs for U.S. municipal and corporate bonds. To determine these costs, we used the investment-grade bonds tracked by the S&P National AMT-Free Municipal Bond Index, the S&P AMT-Free Municipal Series, high-yield bonds tracked by the S&P Municipal Bond High Yield Index, investment-grade corporate bonds tracked by the S&P U.S. Issued Investment Grade Corporate Bond Index, and high-yield corporate bonds tracked by the S&P U.S. Issued High Yield Corporate Bond Index, in conjunction with bond transaction data provided by the Municipal Securities Rulemaking Board (MSRB) and the Financial Industry Regulatory Authority (FINRA). Based on this data, we have determined the average implied transaction cost of municipal bonds since May 2011 and that of corporate bonds since July 2011. This information can help investors compare the cost of buying individual bonds to the cost of investing in bond alternatives, such as mutual funds and exchange-traded funds (ETFs).
Through October 2015, the average implied transaction cost of buying an individual municipal bond of investment-grade quality was 1.21% for retail investors. The average implied transaction cost for high yield municipal bonds was 2.10%. Investment-grade corporate bond transaction costs were lower, at 0.85%, and high-yield corporate bonds had an implied transaction cost of 1.62%.


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Liam Hemsworth named PETA's sexiest vegan
Liam Hemsworth has been named PETA's sexiest vegan.
The 26-year-old actor has been recognised for his hunky looks and plant-based diet by the animal rights organisation, who also bestowed the honour on singer/songwriter Jhené Aiko.
The pair are inheriting their sexiest vegan titles from Liam's girlfriend Miley Cyrus, who he reunited with late last year.
Other past recipients of the accolade include Ellen Page, Kristen Bell, Jared Leto, Jessica Chastain and Russell Brand.
Liam recently explained he changed his diet both for health reasons and because of his love of animals, and although he isn't missing meat, he finds himself longing for dairy.
He said: "I never get cravings for meat. The less I eat meat, the harder it is to imagine eating it. I get cravings for cheese though.
"Every time I'd go to New York, one of my favourite things to do was to go and eat pizza. I've had cheese pizza, like, once in the past four months.
"It was in New York and it was in the first few weeks of being vegan. I was with some friends and we'd had a couple of beers - you walk around and there's pizza everywhere and it's like, 'Ah God, I've gotta have a slice.'"
And the 26-year-old actor's strict diet meant he was "forced" to cook for himself a lot while away shooting 'Independence Day: Resurgence'.
He said: "I do like to cook. I was actually forced to cook a lot more food for myself in New Mexico because you can't really find vegan restaurants there. I did a lot of cooking in high school - I really enjoy it.
"I cook a lot of beans and rice and sweet potatoes and lentils... just the sort of stuff where you can cook a lot of it and throw it in the fridge."


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Martin Davies (17)
Scott Grayson (SSE) (17)
Martyn Thompson (SLA) (19)
Errol Barron (Arch) (17)
Harish Sujan (ABF) (18)
Amy Gaida (Law) (19)
Jane Parker (SSW) (19)
Benjamin Lee (SOM) (17)
Felicia Rabito (SPHTM) (17)
Rosaria Guastella (SCS) (18)
Advisory: To nominate candidates for honorary degrees.
One faculty member from each of the academic constituencies except Newcomb-Tulane College.
Tulane University, New Orleans, LA 70118 504-865-5000 email@example.com


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Here's The List Of Everyone Who's Ever Died From A Cannabis Overdose
As I'm sure you all know, yesterday was 4/20 (the international stoner's holiday) and the internet was awash with weed-related news and information.
Featured Image VIA
But while this plant has time and time again been proven to have serious medicinal and mental benefits, for some reason our government presents it as the root of all evil. The media continues to publish stories about how marijuana and the psychoactive ingredient in it, THC, is a hazard to users' health.
This is obviously an outrageous view to take when far more harmful substances such as alcohol and tobacco are legal. To make this point even clearer, The Huffington Post decided to compile a list of everyone who has ever died solely as the result of a marijuana overdose. Before we go into this, bear in mind that according to the Office for National Statistics, in 2015 there were 8,758 alcohol-related deaths in the UK. That's just in one year.
Comparatively, The Huffington Post discovered that there have been zero recorded incidences of anyone ever dying from a cannabis dose alone. So while there might have been instances of people choking on food or falling over and dying, no one has taken a fatal dose of weed. Ever.
In fact, this is seemingly impossible. Back in 1988, Judge Francis Young, who spoke in favour of changing the laws on marijuana, stated that it would take somewhere between 20,000-40,000 the amount of THC in a regular joint to even approach a lethal dose:
A smoker would theoretically have to consume nearly 1,500 pounds of marijuana within about fifteen minutes to induce a lethal response.
Meanwhile, a recent analysis on cannabis found this figure to be even higher, at an ounce and a half of pure THC, which I think you'll agree would be impossible to ingest.
Image VIA
Now I'm not saying there are literally no dangers in smoking weed – but there are also many, many benefits and the risks are small and mostly only relevant if you're smoking all day every day. When you compare them with the drugs that are legal, it makes absolutely no sense that weed is still not decriminalised.
So Theresa May, if you're reading this, let's make the UK a green country.


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I'm going to share with you my make-up brushes collection!
-Just to point out, I have brought most of these brushes, kindly been given them or they have came free!
I have 2 makes which are the Real Techniques and Seventeen brushes:
~Powder Brush
~Angled Highlighter Brush
~Contour Brush
~Detailer Brush
~Buffing Brush
~Foundation Brush
~Multi-task Brush
~Expert Face Brush
~Fine Liner brush
~Tapered Foundation Brush
~Pointed Foundation Brush
~Base Shadow Brush
^Those ones are all Real Techniques and my favourite one is the Tapered Foundation one, it comes with the Delux set, which I think you can only buy at christmas! It is amazing for bronzing up your face, to give you a nice flawless 'tan' look!
For the Seventeen Brush I own:
~2x Eye Shadow Brush
~Cheek Brush
~Lip Brush
Sorry that this is a short post!
Hope you liked it though
Amy x


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00355295, Extension
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Muscat: Virtually out of reckoning after four losses in a row, a beleaguered Indian football team would look to salvage some pride and put up a decent performance against higher-ranked Oman in a World Cup qualifying round match today.
India are without a win in their World Cup qualifying campaign, even losing to lower-ranked Guam and pressure has been piling on the players and coach Stephen Constantine who took up the job early this year.
In the five-team group, India are yet to open account and have scored just three goals so far. They face an uphill task against Oman, ranked 102nd in FIFA charts as against the 167th of India, who had emerged 2-1 victorious in the first leg in Bangalore.
Constantine's boys would surely be the underdogs in today's match as Oman are high on confidence after holding the continent's highest-ranked team, Iran, to a 1-1 draw on October 8 at home.
On top of that, the Indians are short on practice as they have lost at least a day's training after being stranded in Ashgabat due to non-availability of flights after their 1-2 loss to Turkmenistan.


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I had the pleasure to shoot my first IPSC club match (Level I) up at Port Coquitlam District Hunting and Fishing Club (PCDHFC) on Sunday.
I was one of those rare gloriously sunny west coast spring days. I ran my Smith and Wesson M&P in 9mm. This seemed to be a hot seller in the Production division as I saw quite a few people with the same set up as I.
I placed a respectable 12th out of 20 shooters in the Production division
CTCSupplies gave away two $25 gift certificates as draw prizes, won by TommyC and RobertC. Congrats!
I had some great chats with customers. It's alway a great day when you can chew the fat and shoot with some great guys and gals.
I met some great people including KellyD from Fig.11 Firearms. Fig.11 specializes in firearms safety training and the Canadian Firarms Safety Course and Canadian Restricted Firearms Safety Course. If you are looking to get your PAL/RPAL please give Kelly a shout at email@example.com.
I also had a good chat with BradP from Custom Reoading Services. He's a great guy and a terrific source for reloaded ammunition and components.
Finally, thanks to Dr. Richard Cho and Chung Leung for overseeing a great match!


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Fri 10th Sep 2010 7:00amgoofy_ft's Bob Hall Pier Surf Report
Thigh high and sloppy with some waist high drops. could get better with a changing tide.
View the Bob Hall Pier Surf Report and Forecast. goofy_ft's Reports have received a total of 12040 views.
This report has received 265 views since 10/9/10.


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Top Festival Appearances Ever
Subscribe to KYBB-FM / B102.7 on
When it comes to performances at festivals, Queen's set at 1985's Live Aid tops a Toronto Sun list of the Top Music Festival Performances of All-Time.
"There's no better example of a frontman having the crowd in the palm of his hand than Freddie Mercury at London's Wembley Stadium on July 13th, 1985. In what will go down as one of the greatest live performances in rock history, the English rockers breeze through a six-song greatest hits set in rapid fire succession. Mercury, acting as a choirmaster, leads the 72,000 in attendance in an unforgettable sing-along."
2) Jimi Hendrix at the Monterey Pop Festival in 1967. "We all know him now as one of the most influential guitarists of all time. But in June of 1967, a clearly stoned axeman played to a large — and also very stoned — U.S. crowd for the first time. The reaction on the faces in attendance is priceless — some with their mouths wide open, and some just tripping at the set they're taking in. It all culminates in Hendrix's now-infamous guitar burning finale."
3) The Rolling Stones at Altamont in 1969. "This is memorable for all the wrong reasons. It was dubbed as the official end to the Peace & Love generation. In what was supposed to be the Stones version of Woodstock, the vibe of this free concert turned dark when, earlier on in the day-long rock festival, Jefferson Airplane's lead singer Marty Balin was punched and knocked out by a member of the Hell's Angels, who were inexplicably acting as security for the event (for a reported $500 worth of beer). As the Stones hit the stage, the tension increased, and during 'Under My Thumb,' a scuffle broke out which resulted in the stabbing death of a 18-year-old boy."
4) U2 at Live Aid in London – 1985. "The eager Irish foursome were on the cusp of superstardom, and already had a arena-sized following after the release of 1984's Unforgettable Fire. The powerful showing at Bob Geldof's Ethiopian famine relief benefit put them over the top. Frontman Bono, during an extended version of 'Bad,' pulled a woman out of the crowd and danced with her briefly. The incident caused a frenzy at the front of the stage, but connected with the millions watching. It also caused the band to skip the third scheduled song, 'Pride (In the Name of Love),' because they went over their allotted time."
5) Janis Joplin at the Monterey Pop Festival 1967. "It will be forever known as her coming out party. Joplin, a relative unknown, appeared as a member of Big Brother and The Holding Company, blew everyone away with her intense set, topped by a snarly version of 'Ball & Chain.' The set ends and the camera pans to Mamas & the Papas star Mama Cass mouthing, "Wow, that's really heavy." The band were immediately signed to a contract after the show."


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Join the 'Hybrisil' group to help and get support from people like you.
Hybrisil News
Forget Steroid Shots for Long-Term Relief of Arthritic Knees
Posted 16 May 2017 by Drugs.com
TUESDAY, May 16, 2017 – Steroid shots are a common treatment for arthritic knees. But, a new study says their long-term use is ineffective and may even reduce cartilage. Knee osteoarthritis patients who got steroid injections every three months for two years had no less pain than those taking a placebo treatment. And they had greater loss of cartilage, the rubbery tissue that acts as a cushion between the bones of joints, researchers found. "This research will change how I talk to patients," said Dr. Seth Leopold, a professor of orthopedics and sports medicine at the University of Washington School of Medicine. "I will tell patients that the best information we now have suggests that these injections may not help, and extended use may thin your cartilage," said Leopold, who wasn't involved in the study. "We should only try them in someone who doesn't have alternatives, and we should ... Read more
Related support groups: Osteoarthritis, Chronic Pain, Methylprednisolone, Medrol, Cortisone, Solu-Medrol, Medrol Dosepak, MethylPREDNISolone Dose Pack, Depo-Medrol, Methylprednisolone Topical, A-methapred, Hybrisil, Adlone-40, Duralone, Bupivacaine/lidocaine/triamcinolone, Cortone Acetate, Methacort 40, Dexamethasone/Lidocaine, Med-Jec-40, Dep Medalone 80
Is It Wise to Take a Steroid for a Sore Throat?
Posted 18 Apr 2017 by Drugs.com
TUESDAY, April 18, 2017 – Researchers seeking new sore throat treatments report only modest success with a single dose of a steroid medication. Concerns about growing bacterial resistance to antibiotics have led scientists to look for alternative therapies for sore throat, a common reason for doctor visits. In this new British study, a steroid medication led to improvement in about one-third of patients with sore throat. But, two U.S. physicians said they aren't rushing to prescribe the drugs on the basis of these results. Steroid treatment "might mask a more serious problem. That's really pretty important," said Dr. Robert Centor, a professor of internal medicine at the University of Alabama at Birmingham. "Most sore throats are relatively simple, but some are relatively dangerous, and people can die or end up in the ICU," said Centor, who wasn't involved in the study. "Steroids could ... Read more
Related support groups: Prednisone, Methylprednisolone, Medrol, Dexamethasone, Sore Throat, Decadron, TobraDex, Solu-Medrol, Ciprodex, Medrol Dosepak, MethylPREDNISolone Dose Pack, Maxitrol, Depo-Medrol, Ozurdex, Dexamethasone/Tobramycin, Deltasone, Maxidex, Tobradex ST, Rayos, Decadron Tablets
Steroid Shots Offer No Long-Term Relief for Low-Back Pain
Posted 20 Mar 2017 by Drugs.com
MONDAY, March 20, 2017 – Chronic lower back pain affects millions of Americans. Many try steroid injections to ease their discomfort, but researchers now say this remedy provides only short-term relief. In their study, investigators from France focused on 135 patients with back pain seemingly caused by inflammation between the discs and bones (vertebrae) in the lower spine. The researchers found that a single steroid injection eased pain for one month. After that, however, effectiveness waned. Virtually no difference was seen one year after treatment between patients who did or didn't get the injection. "Our results do not support the wide use of an injection of glucocorticoid in alleviating symptoms in the long term in this condition," said lead researcher Dr. Christelle Nguyen. The findings are consistent with earlier studies, said Nguyen, an assistant professor of physical medicine ... Read more
Related support groups: Pain, Back Pain, Chronic Pain, Methylprednisolone, Sciatica, Herniated Disc, Medrol, Breakthrough Pain, Solu-Medrol, Medrol Dosepak, MethylPREDNISolone Dose Pack, Depo-Medrol, A-methapred, Methylprednisolone Topical, Adlone-40, M-Prednisolone, Hybrisil, Decadron with Xylocaine, Methacort 80, Dexamethasone/Lidocaine
4 Ways to Protect Yourself Against Poison Ivy, Oak and Sumac
Posted 11 Aug 2016 by Drugs.com
THURSDAY, Aug. 11, 2016 – When you're enjoying the great outdoors, be on the lookout for poison ivy, poison oak and poison sumac. The urushiol oil in their sap can cause itching, a red rash and blisters. These symptoms can appear from a few hours to several days after exposure, according to the U.S. Food and Drug Administration. Learn what these plants look like so you can avoid them. The old saying "Leaves of three, let it be" is a helpful reminder for poison ivy and poison oak. But it's not foolproof – the form may vary depending on the type of plant you encounter. Poison sumac, meanwhile, usually has clusters of 7 to 13 leaves, according to the U.S. National Institute for Occupational Safety and Health. If you're working in areas with these plants, wear long sleeves, long pants tucked into boots and impermeable gloves. Wash garden tools and gloves regularly. Wash pets if they may ... Read more
Related support groups: Allergic Reactions, Allergies, Benadryl, Hydroxyzine, Zyrtec, Promethazine, Claritin, Loratadine, Allegra, Diphenhydramine, Phenergan, Vistaril, Cetirizine, Cyproheptadine, Clobetasol, Atarax, Fluocinonide, Fexofenadine, Xyzal, Periactin
Steroid Shot for Hip Pain May Carry Infection Risk If Too Close to Surgery
Posted 2 Mar 2016 by Drugs.com
WEDNESDAY, March 2, 2016 – Patients who've received a steroid injection for hip pain should wait at least three months before having hip replacement surgery, a new study suggests. "The risk of developing an infection after surgery increased significantly in patients who had a hip replacement within three months of receiving a steroid injection," study author Dr. William Schairer, from the Hospital for Special Surgery in New York City, said in a hospital news release. "However, in patients who had a steroid injection and then waited three months or longer to have the surgery, there was no increased risk at all." Steroid injections are widely used to ease pain and inflammation associated with arthritis. These injections can weaken the immune system, which could increase infection risk, the researchers explained. "Hip replacement is a common and safe procedure that relieves pain and ... Read more
Related support groups: Infections, Prednisone, Methylprednisolone, Prednisolone, Hydrocortisone, Medrol, Cortisone, Dexamethasone, Triamcinolone, Hip Replacement, Betamethasone, Budesonide, Decadron, Entocort, Orthopedic Surgery, Solu-Medrol, Entocort EC, Florinef, Fludrocortisone, Cortef
Retail Prices of Dermatology Drugs Skyrocket
Posted 25 Nov 2015 by Drugs.com
WEDNESDAY, Nov. 25, 2015 – Patients using prescription creams, gels, sprays and pills for skin conditions may shell out substantially more at the pharmacy than they did just six years ago, a new study suggests. Between 2009 and 2015, retail prices of brand-name dermatologic drugs rose 401 percent, on average, study authors reported Nov. 25 in JAMA Dermatology. Even generics have succumbed to price inflation, up 279 percent between 2011 and 2014, based on the drugs surveyed. Price increases for skin treatments far outpaced the general inflation rate of 11 percent during the six-year study period, the researchers said. "Cancer drugs were the worst in terms of the numbers" – up 1,240 percent or nearly $11,000 over the six-year study period – primarily because of two medicines, said Dr. Steven Rosenberg, voluntary professor of dermatology at the University of Miami Miller School of ... Read more
Related support groups: Monistat, RID, Monistat 3, Eczema, Monistat 7, Voltaren Gel, Dermatitis, Clobetasol, Contact Dermatitis, Mupirocin, Therapeutic, Bactroban, Efudex, Maintain, Sulfur, Drysol, Hypercare, Epiduo, Retin-A, Fluocinonide
Knee Arthritis: Steroid Shots May Not Help Long-Term, Ozone Injections Promising
Posted 10 Nov 2015 by Drugs.com
TUESDAY, Nov. 10, 2015 – Millions of aging Americans are plagued by arthritic knees, and two new studies offer insight into what might – or might not – help curb the condition. Both studies were presented this week at the American College of Rheumatology's annual meeting in San Francisco. One study found that a popular therapy, steroid drug injections, do nothing to slow progression of osteoarthritis in the knee. This type of treatment is common, but has never been specifically tested, and there are concerns about its safety, according to a team led by Dr. Tim McAlindon, chief of rheumatology at Tufts Medical Center in Boston. His team tracked outcomes for 140 people – mainly overweight white women – with knee arthritis who averaged 58 years of age. The patients received either injections of the steroid triamcinolone hexacetonide, or placebo injections of saline, every three months ... Read more
Related support groups: Prednisone, Osteoarthritis, Testosterone, Methylprednisolone, Prednisolone, AndroGel, Hydrocortisone, Medrol, Cortisone, Dexamethasone, Triamcinolone, Testim, Betamethasone, Axiron, Budesonide, Decadron, Entocort, Kenalog, Androderm, Nasacort
'Tennis Elbow' Usually Heals Without Therapy, Study Finds
Posted 1 Oct 2015 by Drugs.com
THURSDAY, Oct. 1, 2015 – Most people with tennis elbow recover without physical therapy and steroid injections, according to a study by researchers in Norway. "I'm not surprised because that's really been the classic teaching," said Dr. Joshua Dines, an orthopedic surgeon at the Hospital for Special Surgery in New York City. "The number that's often cited is that 90 percent of tennis elbow, golfer's elbow, will get better by the end of the year no matter what you do." Study first author Dr. Morten Olaussen, a specialist in family medicine at the University of Oslo, agreed that the finding was expected. But, he added, "it is interesting to note that after one year, as much as one-third of the patients still reported considerable discomfort." What was surprising, said Olaussen, was that physical therapy was not effective. "It has been shown to be effective in earlier research but then on ... Read more
Related support groups: Methylprednisolone, Medrol, Solu-Medrol, Medrol Dosepak, MethylPREDNISolone Dose Pack, Depo-Medrol, Epicondylitis - Tennis Elbow, A-methapred, Methylprednisolone Topical, Adlone-40, Methacort 40, Methylprednisolone/Neomycin, Med-Jec-40, Dep Medalone 80, Methylcotol, Hybrisil, Medralone, Methylpred DP, Medralone 80, Depopred
Health Tip: Coping With Mosquito Bites
Posted 1 Sep 2015 by Drugs.com
-- Mosquito bites are itchy and annoying, and tough to ignore when they're virtually screaming "scratch me!" The Mayo Clinic recommends: applying calamine lotion, using hydrocortisone cream, applying an ice pack or cold compress, combating a more significant allergic reaction by taking an oral, over-the-counter antihistamine. Read more
Related support groups: Clobetasol, Fluocinonide, Clobex, Desonide, Kenalog, Calamine, Desoximetasone, Elocon, Topicort, Lidex, Cordran, Halog, Cordran Tape, Cloderm, Olux, Cortizone-10, Halobetasol, Beta-Val, Locoid, Vanos
Got a Rash? You Might Be Allergic to Nickel, Dermatologist Says
Posted 20 Aug 2015 by Drugs.com
THURSDAY, Aug. 20, 2015 – Nickel is one of the most common causes of a skin rash that occurs due to contact with an allergen, a dermatologist says. This type of rash, allergic contact dermatitis, can be caused by nickel in jewelry, a patient's diet, nickel in a medical implant or nickel in a medication that's applied to the skin, said Dr. Jennifer Chen, a clinical assistant professor of dermatology at Stanford Hospital and Clinics, in Stanford, Calif. Typically, an allergic reaction to this metal occurs in an area of skin that comes into contact with an item such as a necklace, belt buckle, zipper, eyeglass frames or cellphone. But nickel in foods can cause an allergic reaction that is more widespread on the body. Foods high in nickel include nuts, seeds, chocolate, wheat and rye, Chen said. "Although allergic reactions to dietary nickel are not as common as nickel allergies overall, ... Read more
Related support groups: Allergic Reactions, Allergies, Benadryl, Hydroxyzine, Zyrtec, Promethazine, Claritin, Loratadine, Allegra, Diphenhydramine, Phenergan, Dermatitis, Vistaril, Cetirizine, Cyproheptadine, Clobetasol, Atarax, Fluocinonide, Fexofenadine, Xyzal
Health Tip: Treating Poison Ivy
Posted 25 Jun 2015 by Drugs.com
-- If you've been exposed to poison ivy, a few suggestions can help ease the itch, prevent the rash's spread and reduce your risk of skin infection. The American Academy of Dermatology suggests: Immediately after possible exposure, use soap and lukewarm water to wash the skin. Avoid scrubbing areas already laden with a poison ivy rash. Remove all clothing that could have touched the plant, and wash immediately. Use lukewarm soapy water to wash anything that may have touched the plant, including gardening tools, golf clubs, pet fur or pet leashes. Don't scratch the area, since doing so could lead to a skin infection. Don't rub or remove peeling skin. Soothe itchy skin with a short, lukewarm bath in colloidal oatmeal or baking soda. Then apply calamine lotion or hydrocortisone to help ease itching. Use a cool compress to soothe itchy skin, or take an oral antihistamine. Read more
Related support groups: Benadryl, Hydroxyzine, Zyrtec, Promethazine, Claritin, Loratadine, Allegra, Diphenhydramine, Phenergan, Vistaril, Cetirizine, Cyproheptadine, Clobetasol, Atarax, Fluocinonide, Fexofenadine, Xyzal, Periactin, Chlorpheniramine, Clobex
Poison Ivy's Gonna Get Ya...
Posted 7 Jun 2015 by Drugs.com
SATURDAY, June 6, 2015 – Poison ivy, oak and sumac are common outdoor hazards, but there are a number of ways to prevent exposure and reduce your suffering if you do come into contact with these plants, an expert says. "Millions of Americans every year develop an allergic rash after being exposed, and these poisonous plants are pretty much everywhere in the United States except Alaska and Hawaii," Renee Miller, from the Tennessee Poison Center at Vanderbilt University Medical Center, said in center news release. Oils in the plants' leaves, roots and vines get into the skin almost immediately after contact and bind with proteins, causing an immune system reaction that leads to extreme itchiness. "If there's a risk for exposure, wear long pants, long sleeves, gloves and boots," Miller said. But she noted that rubber gloves won't protect you because the plant oils are soluble in rubber ... Read more
Related support groups: Skin Rash, Benadryl, Hydroxyzine, Zyrtec, Promethazine, Claritin, Loratadine, Allegra, Diphenhydramine, Phenergan, Vistaril, Cetirizine, Cyproheptadine, Clobetasol, Atarax, Fluocinonide, Fexofenadine, Xyzal, Periactin, Chlorpheniramine
Use of Low-Dose Steroid Creams During Pregnancy Won't Affect Baby: Study
Posted 4 Sep 2013 by Drugs.com
WEDNESDAY, Sept. 4 – Women who apply prescription steroid creams such as cortisone to ease a medical issue during pregnancy should not worry that the medication will affect their baby, a new study finds. "Cortisone [creams] can be a very powerful drug for a pregnant woman suffering from a variety of conditions both related and unrelated to pregnancy," noted one expert, Dr. Doris Day, a dermatologist at Lenox Hill Hospital in New York City. "In many cases, there are no substitutes that will relieve the itch and suffering of the conditions, which without proper treatment with cortisone are not only miserable to tolerate but can also lead to permanent scarring," said Day, who was not connected to the new study. Reassuringly, the researchers found that use of corticosteroid gels or creams during pregnancy is not linked to an increased risk for certain complications, including premature ... Read more
Related support groups: Clobetasol, Fluocinonide, Clobex, Desonide, Kenalog, Desoximetasone, Elocon, Topicort, Lidex, Cordran, Halog, Cordran Tape, Olux, Cloderm, Cortizone-10, Halobetasol, Locoid, Betnovate, Beta-Val, Vanos


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Today, I was babysitting my barely-pubescent cousins, and they started talking about giving blowjobs to their "boyfriends". When I got mad at them and told them they shouldn't be thinking of that stuff, they said I was just pissed 'cause I haven't gotten laid. FML
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By bella / Sunday 8 December 2013 01:07 / United States - Edinburg
Today, I had to explain to my daughter that just because it says non-toxic on the crayons, it doesn't mean that you should eat them. She's 16. FML
By no she wasn't high / Monday 2 December 2013 02:50 / United States - Garland
Today, my boyfriend broke up with me but wanted to make sure that we were still friends, so he could still use my Netflix. FML
By unwantedforlife - / Wednesday 20 November 2013 00:19 / United States
Today, my workplace had a big employee photoshoot for an ad campaign. I was there all of 30 seconds before the photographer said, "What the fuck? Look guys, this ain't an ad for facial abortions." He then asked me and another colleague to step out of the shot. FML
By fuggers :/ - / Sunday 24 November 2013 21:38 / United Kingdom - Basingstoke
Today, I had to have a talk with my stalker. After telling him not to snapchat me, not to text me, and that I'm not interested, all he said was "I think persistence is going to be key here." FML
By AshleyRose24 - / Saturday 23 November 2013 08:02 / United States - Fullerton


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Arihant Tools Mfg Co
Address: B/20, Jagdusha Nagar, Paras Darshan Bldg, Golibar Rd, Ghatkopar (w), Mumbai, Maharashtra, 400086
Phone number: 022 - 25151701Post A Recommendation Bookmark this Business Claim this Business
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