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Transforming growth factor-β1 (TGF-β1) is thought to drive fibrogenesis in numerous organ systems. However, we recently established that ectopic expression of TGF-β1 abrogates collagen accumulation via canonical SMAD signaling mechanisms in a shear-induced model of kidney fibrosis. We herein delineate the temporal control of endogenous TGF-β1 signaling that generates sustained synchronous fluctuations in TGF-β1 cascade activation in shear-stimulated proximal tubule epithelial cells (PTECs). During 8-h exposure to physiological shear stress (0.3 dyn/cm2), PTECs experience in situ oscillatory concentrations of active endogenous TGF-β1 that are ∼10-fold greater than those detected under higher stress regimes (2–4 dyn/cm2). The elevated levels of intrinsic TGF-β1 maturation observed under physiological conditions are accompanied by persistent downstream SMAD3 activation. Pathological shear stresses (2 dyn/cm2) first elicit temporal variations in phosphorylated SMAD3 with an apparent period of ∼6 h, whereas even higher stresses (4 dyn/cm2) abolish SMAD3 activation. These divergent patterns of SMAD3 activation are attributed to varying levels of Notch4-dependent phospho-SMAD3 degradation. Depletion of Notch4 in shear-stimulated PTECs eventually increases the levels of active TGF-β1 protein by approximately fivefold, recovers stable SMAD phosphorylation and ubiquitinated SMAD species, and attenuates collagen accumulation. Collectively, these data establish Notch4 as a critical mediator of shear-induced fibrosis and further reinforce the renoprotective effects of canonical TGF-β1 signaling.
- fluid shear
- chronic kidney disease
- tubulointerstitial fibrosis
- frequency-dependent signaling
transforming growth factor-β1 (TGF-β1) is a potent growth factor with many diverse functions. TGF-β1 exerts these pleiotropic effects via a fairly complex signaling cascade, which exhibits multiple points of regulation. Newly synthesized TGF-β1 protein is secreted possessing a latency peptide that effectively blocks activity until properly acted upon by a multitude of molecules (44). Cleaved, active TGF-β1 then binds a type II receptor, which subsequently recruits one of many type I receptors responsible for activating downstream ligands. Canonical signaling involves phosphorylation and activation of the receptor-regulated SMAD transcription factors SMAD2 and/or SMAD3, but MAP kinases can also be stimulated. However, activation of the TGF-β1 cascade does not necessarily initiate a downstream transcriptional response as the levels of both total and phosphorylated SMAD proteins are also subject to tight regulation via ubiquitin-mediated proteosomal degradation. Indeed, a large number of different ubiquitin ligases, such as the SMURF proteins, WWP1 and NEDD4L, are capable of recognizing inactive and/or phosphorylated SMAD proteins and targeting them for destruction (28, 55).
In the kidney, signaling events downstream of TGF-β1 have been hypothesized to drive the development of fibrosis leading to end-stage renal disease and eventual organ failure, potentially by initiating an epithelial-to-mesenchymal transition (EMT) (20). However, we recently demonstrated that not only are EMT and fibrosis mutually exclusive events in sheared proximal tubular epithelial cells (PTECs), but also that ectopic overexpression of TGF-β1 in these cells abrogates EMT-independent matrix accumulation via SMAD-dependent mechanisms (19). Consistent with these observations, recent work by Fragiadaki and colleagues (16) identified dynamic dosage-dependent effects of TGF-β1, whereby low concentrations stimulate collagen synthesis but high concentrations suppress it. Furthermore, we demonstrated that TGF-β1-mediated EMT may be kinetically limited via oscillatory activation of the downstream ERK signaling cascade (5); thus, accumulating evidence suggests that dynamic context-dependent TGF-β1 signaling defines unique cellular responses.
Although it has been discovered that SMAD1, a TGF-β family transcription factor, can exhibit oscillations in response to serum (57), no reports to date have demonstrated sustained oscillations in other TGF-β1 family members. In contrast, the oscillations of the transcription factors NF-κB, p53, and Hes/Her, the most prevalent signaling systems with such dynamics, have been somewhat extensively studied and characterized (37, 42, 53). Additionally, periodic behavior is evident in many important biological processes: the natural circadian rhythms that govern organism behavior (60), the regimented proliferation of the cell cycle (30), and even the complex spatiotemporal expression patterns in systems of genes that drive embryonic development (14, 23). The simplest of these dynamic responses are driven by the presence of a negative feedback loop operating with a delay, although their exact structure can comprise many different network motifs and the resulting output can vary dramatically (18). Because of the inherent interconnectedness of many circuit elements, elucidating key regulatory components of the oscillatory machinery is often both theoretically and experimentally difficult. Here, we discover that a physiologically relevant mechanical stimulus in kidney metabolism, fluid shear stress, induces novel oscillatory variations in both transcriptional expression of TGF-β1 and downstream cascade activation, which are in turn responsible for the regulation of matrix synthesis. Fluid shear stress is an important physical stimulus in kidney homeostasis (25). The physiological magnitudes of shear stress in the proximal tubule range from 0.06 to 0.3 dyn/cm2 (8, 17). Following surgical renal mass reduction, flow in the remaining nephrons has been shown to increase approximately threefold (24). Furthermore, systemic hypertension (33) and diabetes (43) also increase hydrodynamic forces in the kidney. Consequently, understanding the basis for the intrinsic oscillatory TGF-β1 response could highlight complex mechanisms of signal integration in kidney cells and provide new insights into fundamental aspects of renal physiology.
To study the effects of fluid shear on PTECs, an immortalized human PTEC line, HK-2, was subjected to prescribed levels of either physiological (0.3 dyn/cm2) or pathological (2 or 4 dyn/cm2) shear stresses. Because renal damage is the result of chronic insult occurring over decades, we used very high pathological shear stresses (2 or 4 dyn/cm2) to examine fundamental aspects of renal cell function in an experimentally feasible time scale. In other words, we chose the standard approach employed by toxicologists to evaluate the potential toxicity of lifetime exposure of humans to a chemical substance; that is, the investigation of supraphysiological concentrations of the chemical for an experimentally feasible time window.
Prior work in our laboratory demonstrated that collagen accumulation in shear-activated PTECs increases with the magnitude of applied shear stress, further corroborating the critical role of hyperfiltration on the progression of renal fibrosis (19). Moreover, ectopic expression of TGF-β1 evinced distinctly anti-fibrotic effects in shear-stimulated cells (19). To more fully characterize the role of canonical TGF-β1 signaling in shear-induced fibrosis, we examined the time evolution of endogenous TGF-β1-mediated SMAD activation in the absence of an ectopic expression vector. Importantly, we demonstrate shear stress dynamically regulates the SMAD3 signaling axis. Moreover, Notch4, a cell surface receptor important in the well-characterized synchronous oscillatory signaling system of the segmentation clock (12, 20), represents a key regulatory module responsible for the temporal variations of SMAD3 activity observed in shear-stimulated PTECs with direct effects on downstream matrix accumulation. Our data present a novel and important cross-talk between the Notch and TGF-β1 pathways and present new insights regarding the role of these molecules in the progression of renal fibrosis.
Notch4 siRNA and scramble oligonucleotides as well as antibodies specific for p-SMAD2/3, SMAD2/3, and Notch4 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). The proteosomal inhibitor MG-132 and an antibody directed toward β-actin were obtained from Cell Signaling Technology (Danvers, MA). All other reagents for SDS-PAGE experiments were purchased from Bio-Rad Laboratories (Hercules, CA). Mouse monoclonal antibody (mAb) specific to native collagen I was purchased from Abcam (Cambridge, MA). The anti-mouse IgG (H+L) antibody (Alexa Fluor 488 conjugate) as well as Lipofectamine 2000 were acquired through Invitrogen (Carlsbad, CA). The Notch4 intracellular domain (NICD) expression construct was a generous gift from Dr. Mary J. Hendrix at the Children's Hospital of Chicago Research Center (Chicago, IL) (21). Neutralizing α-TGF-β1 antibody was acquired from Genetex (Irvine, CA).
Cell culture and shear stress exposure.
The immortalized human proximal tubular epithelial cells HK-2 (ATCC CRL-2190, Manassas, VA) were grown (37°C in 5% CO2) on untreated glass slides (7.5 × 2.5 cm) in keratinocyte serum-free media (Invitrogen) supplemented with epidermal growth factor (5 ng/ml) and bovine pituitary extract (50 μg/ml). Before shear exposure, cells were incubated for 18 h in media free of exogenous growth factors. Cells were then subjected to a shear stress level of 0.3, 2, or 4 dyn/cm2 for prescribed periods of time in medium lacking growth factor supplements using a streamer flow device (Flexcell International, Hillsborough, NC) (22, 48, 51, 61). In select experiments, the proteosomal inhibitor MG-132 was added to the medium at the indicated concentrations just before the onset of shear exposure.
Transient transfection and reporter assay.
All transfections were carried out employing Lipofectamine 2000 according to the manufacturer's instructions. In RNA interference assays, HK-2 cells were transfected with 100 nM siRNA specific to Notch4 or a scramble control oligonucleotide sequence. For the SMAD reporter experiments, cells were transfected with 5 μg/slide of reporter construct. Transfected cells were allowed to recover for ≥12 h in growth medium and then incubated overnight in shear medium before their exposure to shear or static conditions.
qRT-PCR assays were performed on the iCycler iQ detection system (Bio-Rad) using total RNA (Qiagen RNEasy kit; Qiagen, Valencia, CA), the iScript one-step RT-PCR kit with SYBR green (Bio-Rad), and primers (49, 62). The GenBank accession numbers and forward (F) and reverse (R) primers are as follows: GAPDH (NM_002046.3), F-GGCCTCCAAGGAGTAAGAC, R-AGGGGTCTACATGGCAACT; TGF-β1 (NM_000660.4), F-CAACTCCGGTGACATCAAAA, R-ACGTGGAGCTGTACCAGAAA. The specificity of primers was determined by dissociation curve and 1% agarose electrophoresis (data not shown). GAPDH was used as an internal control. Reaction mixtures were incubated at 50°C for 15 min, followed by 95°C for 5 min, and then 40 PCR cycles were performed with the following temperature profile: 95°C, 15 s; 58°C, 30 s; 68°C, 1 min; and 77°C, 20 s (48, 51, 61). Data were collected at the 77°C, 20-s step to remove possible fluorescent contribution from dimer primers. Gene expression values were normalized to GAPDH.
Western blot analysis.
HK-2 cells from sheared and matched static control specimens were lysed in RIPA buffer (25 mM Tris·HCl, pH 7.6; 150 mM NaCl; 1% Nonidet P-40; 1% sodium deoxycholate; and 0.1% SDS) containing a cocktail of proteinase inhibitors (Pierce Chemical, Rockford, IL) (47, 50). The protein content of the cell lysates was determined using bicinchoninic acid (BCA) protein assay reagent (Pierce Chemical). Total cell lysates (10 μg) were subjected to SDS-PAGE, transferred to a PVDF membrane, and probed with a panel of specific antibodies. β-Actin was used as a loading control. All Western hybridizations were performed at least in triplicate with independent cell samples.
Shear-activated PTECs were washed twice with ice-cold PBS and homogenized in RIPA buffer with complete protease inhibitor mixture for 30 min on ice (9). After centrifugation for 10 min at 16,000 g at 4°C, the supernatant was collected. Protein concentrations were determined by BCA. A total of 300 μg protein was used for each immunoprecipitation experiment. Antibody (2 μg) was added to 0.4-ml protein extracts. The mixtures were rotated at 4°C overnight. Then, 25 μl of protein G agarose beads (Invitrogen) were added to the mixtures, and they were rotated for another 12 h at 4°C. The beads were harvested by centrifugation and washed 5× with RIPA buffer, and 5× with PBS. Finally, samples were boiled for 5 min in PBS to elute bound protein. After centrifugation, the eluants were subjected to immunoblot analysis.
Immunofluorescence staining.
Following the indicated static or shear treatments, HK-2 cells were fixed with 3.7% formaldehyde for 10 min, permeabilized with 0.1% Triton X-100 for 10 min at 4°C, washed 2× with PBS, and incubated in buffer containing 5% BSA/PBS for 60 min at room temperature (RT) (1). Cells were then incubated with a mouse mAb to collagen I overnight at 4°C, washed 2× with PBS, and incubated in buffer containing Alexa Fluor 488-labeled goat anti-mouse IgG for 60 min at RT. Finally, cells were washed 2× with PBS and photographed using a fluorescent microscope (Nikon Eclipse Ti).
Enzyme-linked immunosorbent assay.
Quantification of secreted, active TGF-β1 was performed using a commercially available enzyme-linked immunosorbent assay (ELISA) kit (eBiosciences, San Diego, CA) according to the manufacturer's instructions. Following shear for the indicated periods of time, confluent cell monolayers were carefully scraped into PBS and centrifuged before analysis of supernatant for levels of active TGF-β1. Data were then normalized to the total cell lysate protein concentration of each sample as measured via a BCA assay and corrected for background against static control samples. The experiments were repeated three times in triplicates, and data are shown as means ± SE.
Data represent means ± SE of ≥3 independent experiments. Statistical significance of differences between means was determined by Student's t-test.
Fluid shear stress dynamically regulates TGF-β1-mediated SMAD3 activation.
We recently reported that pathological levels of fluid shear induce renal fibrosis and that ectopic TGF-β1 expression attenuated fibrogenesis in sheared PTECs via SMAD-dependent mechanisms (19). To better understand how fluid shear stress modulates endogenous TGF-β1 signaling and thus potentiates collagen accumulation in PTECs, we quantified TGF-β1 gene expression and assessed activation of the canonical TGF-β1 transcription factor SMAD3 in shear-activated PTECs. Interestingly, we discovered that temporal expression of TGF-β1 mRNA displayed distinct oscillatory variations over the entire range of shear stresses examined herein (Fig. 1). In particular, application of a physiologically relevant shear stress level (0.3 dyn/cm2) rapidly induced low-amplitude bursts of TGF-β1 mRNA expression at 1 and 3 h that subsequently dissipated (Fig. 1A). In contrast, PTECs subjected to a pathological shear stress level of 2 dyn/cm2 exhibited relatively more pronounced TGF-β1 mRNA expression with bursts of transcription separated by ∼4 h (Fig. 1B). At higher shear stresses (4 dyn/cm2), dynamic repression of TGF-β1 transcript expression occurred, although the overall temporal characteristics of the oscillatory response were qualitatively similar to those observed in specimens subjected to 2 dyn/cm2 (Fig. 1C). Immunoblotting assays, using antibodies that recognize both SMAD2 and SMAD3 species, also revealed unique patterns of downstream SMAD3 protein phosphorylation that varied inversely to total protein levels of NICD, the cleaved portion of the activated Notch cell surface receptor responsible for transcriptional induction of Notch-responsive genes (15). Specifically, a polyclonal antibody directed to the COOH terminus region of Notch4 detected a low molecular weight band (∼40 kDa) indicative of cleaved Notch4 intracellular protein resulting from successful receptor-ligand interactions. Additionally, increasing shear stress decreased the persistence of TGF-β1-mediated SMAD3 activation (Fig. 1, D–F). For example, in PTECs subjected to 0.3 dyn/cm2, phosphorylation of SMAD3 protein remained relatively persistent over the time period examined, disappearing only ∼7 h when a marked increase in the protein levels of NICD occurred (Fig. 1D). To the contrary, SMAD3 phosphoprotein displayed more transient oscillatory dynamics in specimens subjected to 2 dyn/cm2 (Fig. 1E). Of note, protein levels of the intracellular domain of Notch4 varied anti-phase to these fluctuations in SMAD3 activation, which reinforced its role as a potential negative regulator of the TGF-β1 cascade (Fig. 1E). Finally, a shear stress level of 4 dyn/cm2 resulted in negligible amounts of both phosphorylated and total SMAD3 protein, while NICD reached dramatic and sustained levels under these conditions (Fig. 1F). In light of our recent observations establishing the anti-fibrotic effects of SMAD-dependent signaling (19), we hypothesized that Notch4 acts as a critical mediator of collagen accumulation in shear-activated PTECs by interfering with TGF-β1-mediated SMAD activation.
Due to the complex nature of the TGF-β1 signaling pathway, Notch4-dependent regulation may occur over multiple levels of TGF-β1 signaling. Shear-stimulated cleavage of Notch4 and liberation of the intracellular domain may decrease extracellular activation of latent TGF-β1 and/or directly modulate the stability of downstream intracellular signaling components. We first quantified the levels of active TGF-β1 in shear-stimulated PTECs via an ELISA directed specifically toward mature TGF-β1 protein. Overall, a marked decrease in the levels of accumulating mature TGF-β1 was noted with increasing levels of fluid shear (Fig. 2, A, B, C). These data suggest that, to some degree, the observed variations in canonical signaling may be due to tight regulatory control of TGF-β1 latency complex processing. Interestingly, although PTECs exposed to high shear stresses (4 dyn/cm2) experienced repression of TGF-β1 gene expression and displayed less overall SMAD3 protein (Fig. 1, C and F), active TGF-β1 was still detected in the cell supernatant albeit at low levels (Fig. 2C). This apparent paradox suggests that PTECs under high shear exposure may access preexisting pools of latent TGF-β1 in the extracellular milieu. Furthermore, PTECs subjected to a shear stress level of 4 dyn/cm2 displayed more significant bursts of active TGF-β1 (Fig. 2C) compared with specimens exposed to 2 dyn/cm2 (Fig. 2B), although overall SMAD3 activation declined (compare Fig. 1, E and F). To confirm that the observed SMAD3 phosphorylation was solely a result of TGF-β1-mediated signaling events, confluent PTECs were sheared in the presence or absence of a neutralizing TGF-β1 antibody and assessed for phosphorylated SMAD3 protein. Blocking binding of TGF-β1 to its receptor successfully abolished SMAD3 protein phosphorylation (Fig. 2D). These observations link expression of active TGF-β1 directly to downstream SMAD3 activation. Collectively, our data clearly demonstrate that PTECs exert precise temporal control over the activation of endogenous TGF-β1.
Shear-mediated fibrosis requires Notch4-dependent degradation of phosphorylated SMAD3.
To establish whether ligand-mediated Notch4 cleavage and successful release of the intracellular domain regulate SMAD3 activation, PTECs transfected with a NICD expression construct (21) were subjected to a shear stress level of 0.3 dyn/cm2. This molecular intervention resulted in a dramatic decline in both total and phosphorylated SMAD3 protein levels (Fig. 3A), which is consistent with Notch-mediated suppression. We next wished to elucidate the mechanism by which Notch4 regulates the TGF-β1 cascade. To this end, confluent PTECs were transfected with either Notch4 siRNA or control oligonucleotides and subjected to static or shear stress conditions (4 dyn/cm2). Not only did Notch4 knockdown remove transcriptional repression of TGF-β1 (Fig. 3B), but it also recovered persistent SMAD3 phosphorylation in PTECs sheared at 4 dyn/cm2 (Fig. 3C). This response closely resembles our observations at the physiologically relevant shear stress regime where cleaved NICD was relatively absent (Fig. 1D). Importantly, even complete depletion of the NICD did not alter the oscillatory expression profile of the TGF-β1 transcript (Fig. 3B). These data point to the potential existence of additional oscillatory pathways controlling TGF-β1 signaling. Since pulsatile expression of TGF-β1 mRNA resulted in a concomitant plateau of SMAD phosphorylation (Fig. 3, B and C), we examined whether NICD directly increased the levels of active TGF-β1 protein or whether Notch4 perhaps controlled downstream SMAD degradation. To this end, we examined the levels of circulating active TGF-β1 in Notch4 knockdown samples (Fig. 3D). Although early during shear exposure depletion of NICD had little to no effect, active, mature TGF-β1 levels surged dramatically ∼6 h (Fig. 3D). We also performed an immunoprecipitation assay using an anti-SMAD3 antibody along with cell lysates from PTECs subjected to high stress (4 dyn/cm2) in the presence of the proteosome inhibitor MG-132, vehicle control transfection, or Notch4 siRNA transfection. Both MG-132 and Notch4 depletion successfully recovered SMAD3 and ubiquitinated SMAD3 species in shear-activated PTECs (Fig. 3E). Additional immunoprecipitations performed employing an IgG control clearly revealed that the observed immunopurified species did not reflect nonspecific binding contaminants (data not shown). These findings establish that liberation of the NICD plays an important role in regulating the levels of active TGF-β1 protein and degradation of downstream SMAD3 protein. Furthermore, given the antagonism of SMAD-dependent signaling to extracellular collagen deposition (19), these data suggest Notch4 may be a critical mediator of shear-induced fibrosis.
We next wanted to examine whether these results extend to the shear stress level of 2 dyn/cm2, where SMAD ubiquitination was less dramatically apparent. In particular, we wished to determine whether targeted degradation of phosphorylated SMAD3 could explain the temporal variations of SMAD3 activation observed in PTECs subjected to this stress level. Indeed, in the presence of the proteosomal inhibitor MG-132, shear-activated PTECs (2 dyn/cm2) exhibited persistent SMAD3 activation that accumulated with time (Fig. 4A). Despite sporadic degradation of the phosphorylated SMAD3 species in PTECs exposed to 2 dyn/cm2, total SMAD3 remained relatively stable. This may reflect specific targeting or preference of degradation for the phosphorylated form as previously described in the literature (12, 38). Total SMAD protein may disappear at higher stresses, then, due to an overall higher ratio of the activated SMAD species to unphosphorylated form. Together, these findings identify ubiquitination as a key regulatory mechanism contributing to the dynamic responses of SMAD3 activity in shear-activated PTECs.
Following SMAD3 phosphorylation by TGF-β1 receptor complexes, the transcription factor partners with SMAD4 and translocates to the nucleus where numerous proteins and cofactors are capable of binding SMAD molecules and regulating downstream transcriptional activation (40, 56). To assess whether the variations in SMAD3 phosphorylation detected via Western blotting correlate with direct differences in SMAD transcriptional activity, we transfected confluent monolayers of PTECs with a green fluorescent protein (GFP) reporter construct under the control of SMAD2/3 transcriptional elements. Transfected cells were subsequently subjected to a shear stress level of 2 dyn/cm2 for prescribed periods of time, and GFP expression levels were evaluated via fluorescence microscopy (Fig. 4B). Our results confirmed that SMAD transcriptional activity in shear-stimulated PTECs directly corresponded to increased SMAD3 phosphorylation as detected via Western blotting (Fig. 1E). More importantly, SMAD transcriptional activity in each individual cell fluctuated in concert with the surrounding cells. For such synchrony to occur, the cells themselves must be coupled either directly, as is the case in voltage-controlled synchronous Ca2+ oscillations (26, 54), or indirectly entrained via a "pacemaker" pathway as with the segmentation clock in vertebrates (14, 23). Thus, these data support the involvement of multiple pathways, such as Notch, in the complex temporal regulation of TGF-β1 signaling in sheared PTECs.
To exclude the possibility that Notch4 knockdown results in asynchronous cellular fluctuations that cumulatively hide a persistent oscillatory signal in a Western blot, PTECs were cotransfected with Notch4 siRNA and a GFP SMAD2/3 transcriptional reporter before exposure to 2 dyn/cm2. Our results indicate that Notch4 depletion resulted in persistent SMAD transcriptional activity within the confluent PTEC monolayer and no loss of synchrony was observed (Fig. 4C). Thus, knockdown of Notch4 and inhibition of the intracellular domain fundamentally alter the cyclic dynamics of TGF-β1-mediated SMAD signaling. Since our previous work (19) associated canonical TGF-β1 signaling with an overall anti-fibrotic response, the consequences of Notch4 knockdown on type I collagen deposition were also examined. While sheared PTECs transfected with scramble control displayed a diffuse fluorescent staining pattern consistent with extracellular collagen accumulation, shear-activated PTECs transfected with Notch4 siRNA exhibited primarily intracellular staining with clear demarcation of individual cells as assessed via immunofluorescence (Fig. 4D). The effects of Notch4 signaling on collagen expression were also examined via qRT-PCR analysis of mRNA from PTECs transfected with a NICD construct and subjected to a shear stress level of 2 dyn/cm2. Cumulatively, these data conclusively identify Notch4 as a key regulator of TGF-β1 signaling and a potential therapeutic target in renal fibrosis (Fig. 5).
Our findings clearly establish that exposure to pathological shear stresses results in diminished accumulation of active TGF-β1 in PTECs and, more importantly, inhibits canonical TGF-β1-mediated signaling via Notch4-dependent degradation of phosphorylated SMAD3 protein. Furthermore, we identify oscillatory variations in both the levels of TGF-β1 transcript and the availability of mature signaling factor at least partly due to Notch4 regulation. Although depletion of NICD results in uniform SMAD activation in shear-activated PTECs, it does not alter the fundamental pulsatile pattern of TGF-β1 gene expression, suggesting that other oscillating pathways may also regulate TGF-β1 signaling. Given the numerous reports that establish that TGF-β1 is capable of inducing its own transcription (58, 59), it is possible that recovery of TGF-β1 expression in Notch4 knockdown samples is partly due to recovery of active SMAD3 protein. The potential conversion of a plateau of SMAD phosphorylation into a spurt of TGF-β1 gene transcription could be accomplished through the additional temporal regulation of promoter binding via the partnering of phosphorylated SMAD3 with any one of a number of cofactors such as the AP-1, Ets, forkhead, and homeobox families (32). Similarly, the role of Notch4 in regulating the levels of active TGF-β1 secreted by shear-stimulated PTECs could be mediated via multiple different mechanisms. Integrins (43) and proteases, such as plasmin, thrombospondin, or matrix metalloproteinases, are each capable of liberating active TGF-β1 from the latency peptide (29), and thus they represent potential targets of Notch4 action. Additionally, a host of different molecules have also been shown to target SMAD3 for degradation that could lie downstream of ligand-mediated Notch4 cleavage of the intracellular domain (28, 55). In light of our recent observations showing that TGF-β1 exerts anti-fibrotic effects (19), our new data present striking evidence suggesting that dynamic temporal regulation of the TGF-β1-Notch4 signaling axis is a critical component of shear-induced fibrosis.
Although many other studies have shown a fibrogenic role for TGF-β1 (27, 35), we and others challenge the simplicity of this finding. For example, recent work has suggested overexpression of latent TGF-β1 exerts renoprotective effects, although the authors link these consequences to overall decreases in downstream TGF-β1 signaling via a negative feedback (52). A phase I/II trial of anti-TGF-β1 therapy in systemic sclerosis failed to note any efficacy in halting the development of fibrotic lesions (13). Perhaps TGF-β1 itself is a pleiotropic molecule capable of exerting both anti-fibrotic and profibrotic effects with the overall phenotypic consequence determined by the temporal characteristics of the stimulus. Along these lines, recent work has discovered that low doses of TGF-β1 stimulate expression of collagen while high doses of TGF-β1 trigger inhibition of collagen I via the negative transcriptional regulator CUX1 (16). These observations are in concert with our current data, wherein physiological shear stress is beneficial by inducing larger pulses of active TGF-β1 and persistent downstream SMAD3 activation, whereas pathological shear stresses result in smaller and less persistent TGF-β1 activity. Our ELISA data show distinct temporal profiles of low concentrations of active TGF-β1 at both 2.0 and 4.0 dyn/cm2, thereby suggesting frequency modulation as a potential regulatory mechanism. Furthermore, others have demonstrated this phenomenon whereby differences in pulse frequency of TNF-α result in observable differences in transcriptional activation of different classes of downstream genes (5). Finally, it is also possible that the involvement of an as of yet unidentified pathway acts in concert with TGF-β1 and directly modulates the signaling cascade. The end results of TGF-β1 signal transduction, or even the predominant signaling mechanism, whether it be via SMAD3 or some specific kinase cascade, could be entirely context dependent. There is already extensive evidence of the context-dependent, contradictory roles TGF-β1 plays in cellular physiology in the literature (2, 10).
Since oscillatory fluctuations of TGF-β1-mediated SMAD3 activity in shear-activated PTECs are dependent on variations in the levels of cleaved NICD, it is possible that our observations reflect an incarnation of the segmentation clock. In developing vertebrate embryos, oscillations in Notch signaling arise via feedback from the induction of inhibitory genes such as lunatic fringe (11). Coupled with similar oscillations in the WNT and FGF pathways, a synchronous periodic expression system develops whose spatiotemporal variations allow for precise tissue organization (14, 23). Because the TGF-β1 family of growth factors serves important roles during development (3, 36), it is not surprising for there to be significant cross-talk between these pathways. Indeed, Notch1 has been implicated in both tubulointerstitial fibrosis and EMT (4, 39), while both Notch2 and Notch3 have been shown to antagonize the effects of TGF-β1 (31, 41). The activated intracellular domain of Notch4, in particular, has been shown to bind to the receptor-regulated SMAD3 and block TGF-β1 transcriptional activity (46). Our results provide a new mode of interaction between these two important pathways and further highlight the importance of temporal control exerted by the Notch pathway. Although there is less evidence for its involvement, the circadian clock, the rhythmic expression of genes that control our day-night cycles, has also been shown to control periodic expression of a number of different genes in the kidney (45). Disruption of the natural period of these oscillators even disrupts blood pressure profiles and results in organ damage (34). At the very least, these studies further reinforce the importance of cyclic signaling events to kidney homeostasis.
Taken together, our data indicate that the sole focus on TGF-β1 as a therapeutic target in renal fibrosis may be inadequate. It is possible that some other growth factor or pathway occupies a more fundamental role in driving shear-induced fibrosis. The family of platelet-derived growth factor (PDGF) proteins is typically associated with stimulating growth of myofibroblast-like cells (6), and inhibition of the PDGF-D isoform, in particular, is capable of preventing tubulointerstitial fibrosis in a rat model (7). Finally, we identify synchronous oscillatory dynamics in the TGF-β1 signaling cascade. This discovery suggests that TGF-β1 signaling is subject to complex modes of behavior with potentially divergent downstream consequences. To this end, only a truly systems level analysis, as opposed to an approach focusing on one individual molecule operating on narrow time scales, could fully understand relevant signaling phenomena.
This work was supported by grants from the National Cancer Institute (U54-CA143868) and Kleberg Foundation.
No conflicts of interest, financial or otherwise, are declared by the author(s).
Author contributions: B.M.G. and K.K. conception and design of research; B.M.G. performed experiments; B.M.G. analyzed data; B.M.G. and K.K. interpreted results of experiments; B.M.G. prepared figures; B.M.G. and K.K. drafted manuscript; B.M.G. and K.K. edited and revised manuscript; B.M.G. and K.K. approved final version of manuscript.
- Copyright © 2013 the American Physiological Society
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The Business Innovation stream
is for people who want to own and manage a new or existing business in...
Significant Investor Stream
If you apply for the
Significant Investor stream
, you must be prepared to invest at least AUD 5 million in at least one of the following "complying investments": Commonwealth, state or territory government bonds Certain managed funds that have a mandate for investing in one...
Business Talent Australian Visa
Business Talent Australian Visa
has two streams. The Significant Business History stream is for affluent owners or part owners of a business who want to have a major management role in a new or existing business in Australia. The Venture Capital Entrepreneur stream is for...
Australian Business Visa
Australia Business visa
is the most famous method of immigration that is ideal for those who have proved their successful business skills in their current place of residence and who nurture the aim of taking their success further into the country of Australia and run the business...
GSM Visas NOT Subject to Priority Processing
Currently, the following Australian Visas are NOT subject to priority processing: Skilled - Recognised Graduate Subclass 476 Skilled - Designated Area - Sponsored (Residence) Subclass 883 Skilled - Regional Subclass 887. ...
Investor Stream
If you apply for the
Investor stream
, you must: be younger than 55 years of age, although a state or territory can waive this requirement if your proposed business or investment activity will be of exceptional economic benefit to the relevant region. score at least 65 on the...
List of Skilled Occupations
Please don't hesitate to check your chances even if you are not on the list, as there are many other professions, as we have just listed the main ones and the list changes from time to time. Your personal data will be examined individually and there might be other factors that...
Processing Priority for Business Skills (Provisional) Visa Applications
1. applications from people who are sponsored by the Commonwealth or a State/Territory government 2. all other valid applications are to be processed in the order in which they are received. ...
Requirements for Working Holiday Visa (WHV) in Australia
Australia has quickly become one of the top destinations for immigrants because of its growing economy and high standard of education. To encourage the exchange of cultures and to develop stronger relationships between countries, the Australian Government has made it possible to live in the country...
Business Innovation Stream
If you are in the
Business Innovation
stream, you must: be younger than 55 years of age, although a state or territory can waive this requirement if your proposed business will be of exceptional economic benefit to the relevant region score at least 65 on the Innovation Points...
© 2017 Synch1


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PSA Wide Bandwidth Digitizer External Calibration Wizard
The PSA Wide Bandwidth Digitizer External Calibration Wizard (PSA Option 235) is a Microsoft Windows based program used with the PSA Series Spectrum Analyzers that have Option 122 80 MHz Bandwidth Digitizer installed.
How to Use the Wizard
For detailed guidance on use of the Wizard select "Using Extended Calibration Software for Wide Bandwidth Measurements" AN 1443.
|Release Date
|Version Description
|File available for download...
How to Download
- Click the Download button.
- Save the file to your PC.
How to Install
- Close all open or running applications by clicking Alt + Tab, selecting the running program and close.
- Go to the location where you saved the PSA firmware package, double click on the executable file to install the program.
To Configure the Calibration Wizard
- Start the Wizard and select the Help button.
- Click on Configuring your Sytem the Table of Contents.
- Does Not Require a License
Installs on:
PC / Instrument
Operating System
- Windows XP
- Windows 2000
- Microsoft Windows based personal computer
- Windows 2000, Windows XP Professional
Supported Instruments
- E4440A, E4443A, E4445A, E4446A,E4447A, E4448A with Option 122 installed


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Forgot your password? Enter the email address you signed up with and we will send you a link to create a new password.
Your password reset has been submitted to the address provided.
Standard character mark
Direct mail advertising; direct marketing advertising for others; advertising services, namely providing corporate and brand identity for others; advertising, namely creating corporate logos for others
Business in Houston, TX


text:
A TWO minutes silence will be held in memory of two Southampton firefighters who tragically lost their lives tackling a tower block blaze last year.
Firefighters Alan Bannon, 38, and Jim Shears, 35, died after being overcome by extreme heat while tackling a serious fire at Shirley Towers on April 6 last year.
All residents of the 15-storey block – including one heavily pregnant woman – were either evacuated or led to safety.
Staff across Hampshire Fire and Rescue Service will mark the one year anniversary of the tragedy by observing the silence at 11am on Wednesday next week and flying flags at half mast.
The fire service drew up the plans with Alan and Jim's families and members of Red Watch at St Mary's Fire Station where the pair served.
Members of Red Watch will lay flowers at a memorial stone at fire headquarters in Eastleigh to remember their colleagues.
Chief officer John Bonney said: "As we move towards the one year anniversary of the tragic fire, we pause to reflect on the year and pay tribute to our lost colleagues.
"Since the incident we have moved forward as a Service and focus on new challenges, but Jim and Alan and their families will forever remain in our thoughts."
The fire service has opened online books of condolence for members of the public to pay their respects.


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The frontier may be geographically rugged but there's no dearth of goodwill amongst its elected representatives when it comes to power.
To ensure that wealth and seats stay within the incumbent political elite, elected representatives from the Federally Administered Tribal Areas (Fata) have decided to either propose names of their family members for the upcoming Senate elections, or make hay otherwise.
The procedure
The Senate has 8 representatives from Fata, four of whom will retire in March 2012. The four new members for these vacant seats are voted for by the 12 representatives of Fata in the lower house of parliament.
The National Assembly has 11 Fata members at present, since elections were not held in South Waziristan due to the ongoing insurgency.
Of these 11 members, eight have paired themselves based on mutual understanding and will pay Rs150 million per pair into a pool that will eventually be divided amongst the existing 11 members, sources told The Express Tribune.
The arrangement
The pairs include Muneer Orakzai with Mohammad Kamran Khan; Sajid Toori with Jawad Hussain; Malik Bilal Rehman with Shaukatullah Khan; and Noorul Haq Qadri with Hameedullah Jan Afridi.
Each pair will nominate one person, which could be one of either lawmakers, or their family member, for the four vacant seats. Both lawmakers, however, will pay Rs75 million for being part of this arrangement, making a total pool of Rs600 million which will be divided amongst nine lawmakers.
Zafar Baig Bhittani, the ninth lawmaker of the group, has expressed no interest in making any nominations for the Senate seats and wants to simply receive a payout from the divisible pool for being a silent part of the arrangement.
These nine lawmakers together have offered Rs5 million each to Akhunzada Chattan of Bajaur Agency and Abdul Malik Wazir from South Waziristan Agency for not proposing any candidates either.
The allotments
Muneer Orakzai, leader of the Fata parliamentarians, has already been allotted a ticket in his pair. From two other pairs, Shaukatullah Khan and Noorul Haq Qadri have proposed names of their brothers, Hidayatullah and Abdul Malik, respectively.
When approached, Muhammad Kamran Khan, the lawmaker from North Waziristan, refused to comment on the issue and said that "none of it has been finalised as yet".
Other lawmakers could not be reached for their version, despite repeated attempts.
According to sources, the deal is set to be finalised by February 9.
Published in The Express Tribune, February 9th, 2012.


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Sofia Vergara Long Wavy Casual Hairstyle - Medium Brunette (Chocolate) Hair Color
Description(Added: 3rd March 2011 )
These luscious locks flow over the shoulders showing off the layers cut around the sides and back to lighten the length and enhance the movement of the soft waves. This is a low-fuss hairstyle perfect for those with natural long locks.
- Hair Category: Casual
- Hair Length: Long
- Hair Elasticity: Wavy
- Gender: Women
- Face Shape: Oval, Oblong, Square, and Diamond
- Hair Density: Medium/Thick
- Hair Texture: Medium/Coarse
- Age: Under 21, 21 - 30, 31 - 40, and 41 - 50
- Height: Any
- Weight: Thin/Average/Large
- Glasses: Suits with and without
- Styling Time: 30 minutes
- Products: Mousse, Smoothing Shine, and Hair Spray
- Styling Tip: Apply your smoothing shine to the top sections and then work it all the way down to the ends to soften this style.


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I'm new yes and I need to get my legs for this place, but I've noticed that even though I uploaded a profile picture it doesn't appear in my posts. I tried to reload the picture but the option to change my profile picture has been elusive. No idea how to change it now, don't quite remember how I did it in the first place either.
Do profile pictures have a delay for approval before they appear?


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A local woman is being honored at the 2014 Martin Luther King Jr. Prayer Breakfast for her contributions to the community.
Kathy Dickinson is one of five adults in the area that will be recognized for their dedication to improving their surroundings. Dickinson and her daughter started the non-profit organization Zanesville Community of Voices after a tragic loss of a local youth due to violence.
"We started two years ago because there was a death in the community of a young man by the name of Donte Wolford and that's why we started just because the crime and in the community," said Dickinson.
The group aims to keep children off the streets and provide them with fun and educational activities that inspire change.
"My most enjoyable thing that we have done is a summer lunch program and it ran for eight weeks in the summer and we took the kids different places around the community," said Dickinson.
She says if you are interested becoming a member of the Zanesville Community of Voices it is easy and anyone is welcome to join. The membership is $5 per person and they have meetings on the last Tuesday of every month at 502 Market Street. The MLK Jr. Prayer Breakfast will take place Monday morning at North Terrace Church of Christ at 9 a.m.


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Iggmark Form is a one man design studio i Stockholm. I specialize in visual identities and digital design.
I'm currently on the look out for new and exciting projects and collaborations. If you think that your project could benefit from my participation in any way, I'll be happy to hear from you. Meanwhile, have a look at my work.


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Fast Food, Living Fast
by Anita NelsonIt's just a job. Grass grows, birds fly, waves pound the sand.
-- Muhammad Ali
One ounce of mustard. A handful of chopped lettuce. Three tomatoes. Four pickles. Sprinkle diced onions. Peel a slice of cheese, place it neatly over the bottom bun, slap it atop the patty on the grill, and slide it on top of the veggies. Toast another bun. Light mustard. Chopped lettuce. Three tomatoes. Four pickles? Sprinkle onions. Cheese, slap, boom, slide. Mustard. Lettuce. Tomatoes. Pickles. Onions. Oops, customer said, "No pickles." Peel off pickles and discard. Slap, boom, pow. Mustard. Lettuce. Tomatoes. Pickles. Slap, slide. Mustard, pickles, tomatoes, pickles, onions, slap. Mustard, pickles, tomatoes, onions, mustard, pickles, tomatoes, onions, slap! Slap! Pow! STOP! Okay, pause. Rewind a bit -- nope, a little more... more... more -- okay, you know what? Just go all the way back to the beginning.
... you have no formal training in the fast food industry. In fact, you have no work experience, but that's okay, you think; after all, it's only fast food . Sixteen-year-olds sign up for this all the time, and you've got a nifty high school GPA and are pretty cunning in the street-smart department, if you do say so yourself.
It's your first day on the job. Other than the uniform you bought and the mandatory orientation (during which the training manager effectively wasted an hour reiterating the concept of "Sell! Bam! We're running a business!"), you have no formal training in the fast food industry. In fact, you have no work experience, but that's okay, you think; after all, it's only fast food. Sixteen-year-olds sign up for this all the time, and you've got a nifty high school GPA and are pretty cunning in the street-smart department, if you do say so yourself.
Olivia, the overweight manager, greets you with twinkling eyes - which you will only later identify as the same gleam a lion's eyes make before coming in for the kill - so you accept her kind face with gratitude and get introduced to the crew and kitchen. This is Leslie, and that's the freezer. The meats are kept there. Hi Emily, Barbara, Jacob, and that scrawny kid whose name you don't quite catch. Refrigerator, vegetables. Stock. Lauren, Omar. Office, drive-through station. Eric. It's all one long unassorted list that you long to make sense of, but time is ticking fast, so you skip after Olivia to commit more names and places to non-memory. It can't be that important anyway; you're quitting in three months.
Five minutes later, you're wearing a headset and Lauren has you filling up drinks that drive-through customers order while she answers customers on her headset and at the window, calculates correct change, stuffs fries into cartons, and hands out food orders. Your mind cries: This has got to be the easiest job you've ever done. No wonder it's paid minimum wage. What you don't know is that tomorrow, you'll be the one working the register Lauren has today; tomorrow, there won't be a Lauren doing everything for you. If you knew better, you'd be choking on spit right about now.
Fifteen minutes before your three-hour shift will end, Lauren hands you a mop and bucket, and you look at her with the slightest tinge of disgust. Isn't this her job? Never mind the fact that you'll be doing this tomorrow; that can't be used against you, not when you don't know it yet. And so your face morphs back into a smile and with that complacent nature you've always prided in yourself, you fist the mop and nearly knock the bucket over as you begin to work your way through the greasy, greasy floor.
Fast forward two and a half months. You now know better than to try to beat Jacob's four second record for building hamburgers or to interrupt Annette when she's working at the drive-through station. Omar's just been promoted to team leader, and Lauren will be doing the same in February. Michelle's the new face this week, but she won't last long1, so you don't even bother to befriend her. Olivia sends her over to Anita, who sighs. Again. She's trained four new 'uns this month already; no one has lasted more than two weeks.
Four months in, you work your first graveyard shift. You remember telling the gang you'd never work a night shift -- you need to sleep and do homework and breathe, guys -- but Matthew used that puppy-dog face and really, you just need the money. It's not the most ideal situation; Daniel, the night manager and the only manager whose butt isn't well acquainted with the chair in the back office, has the day off. You don't know how to make fast food breakfast in any form, so the only reason you decided to do this was because -- wait, why'd you decide to do this again?
It's too late to back out now, so you work until 1:00 AM, at which time the substitute night manager tells you to clock out. What? She tells you labor is costly, meaning there have not been enough sales to pay workers and maintain the company's 80% profit 2, so someone has to clock out, and everyone else has already done this, so it's only fair you work off the clock for a while too. You're outnumbered, two lazy bums to one timid you; so you clock out for an hour.
The drunk rush is punctual, arriving right as the second hand reaches the sixty-second mark and the two o' clock hour begins. Fewer workers means more chores for you, especially because managers do no more than criticize your work. You are okay with multi-tasking because it gives you excuses to run away from customers. These vile, nocturnal male customers hit on you, attempt to grope you, try to bribe you for your phone number, and suggest activities they'd like to partake in with you beside them, wink, wink. No wonder girls beg to work in the back on nights. How to escape? The night manager watches the camera, but does nothing.
Half an hour in, you're instructed to clean up some dried blood in the men's bathroom; it's from another customer brawl, this time between two drunk brothers smitten over the same girl. Afterward, you wash dishes, sweep and mop, de-salt the fries machine, and remake fresh iced teas for the morning shift.
Half an hour in, you're instructed to clean up some dried blood in the men's bathroom; it's from another customer brawl, this time between two drunk brothers smitten over the same girl. Afterward, you wash dishes, sweep and mop, de-salt the fries machine, and remake fresh iced teas for the morning shift. Your fellow co-worker scrapes the grill and clocks out. No more. You're quitting. Today.
Six months later, Anthony clocks out just as you clock in. He eagerly does a final count of the money in his register and takes off down the street -- hey! he's supposed to wait for you to count yours! -- while your daydreaming eyes cast a final disapproving glance on his vanishing silhouette. Ding! That headset is going to be the death of you. Before you answer -- let those pesky customers wait -- you show the new trainee the onion rings under the fries in the refrigerator in the back. Finally, you turn up your headset speaker and adopt a professional attitude. "Thank you for choosing Onion Rings. This is Annette; how may I help you?"
Credit: Creative Commons License
The new trainee erupts into nervous laughter, and soon the entire kitchen is spilling tears of joy, and jeez, what's so funny? Olivia calls you into the office, and when you finally greet her twinkling eyes, you cower.
"Do you think working here is a joke?" Yes, mostly because I've never seen you get up once from that chair. Could you put on any more weight? Jeez, I was supposed to quit last winter! You don't actually say any of that, not when you need the job so desperately to pay rent, and especially not when her voice mimics the tone she used to fire Omar. If there's one thing you've learned, however, it's that you can't let the lazy bum boss you around -- especially not after six mystery shoppers3 have awarded you perfect scores - so you stick your nose in the air and trot back to your station. Later, while Jacob is rolling up a couple of green leaves and drawing a lighter to his roll -- you refuse to read into the implications of his actions - he teases you about the "onion ring" slip-up. You smirk.
Ashley drops a patty on the floor, but the company's low on large meat, and she's not willing to be blamed. Swoop! The manager arrives, but the victimized meat already lies tucked between two buns, ready to be wrapped and served, ...
Ashley clocks in late for her morning shift. Again. She should know better than to take advantage of you, especially when you work thirteen-hour shifts minimum a day. No one else cleans during the night shift like you do, so she better start appreciating what she's got. Fast. Stupid team leaders with their stupid authority. You're quitting. Today. It's an oddly familiar sentiment that you haven't acted upon in the past months, and you've no doubt that its more frequent appearances will do little to change the future. Ashley drops a patty on the floor, but the company's low on large meat, and she's not willing to be blamed. Swoop! The manager arrives, but the victimized meat already lies tucked between two buns, ready to be wrapped and served, never to be suspected of touching a floor. The first time it happened, you felt shocked, but now, it seems routine. God made dirt, and dirt don't hurt. You, of course, don't condone Ashley's actions; then again, you're not willing to pay for an accidental slip up, either. Whatever.
Finally. It's been nearly a year. You've lived through the federal raise in minimum wage, even though you're still sore about how it eclipsed that $0.50 raise your boss finally gave you, you've fought off the night rush, you've outlived most employees here, and really, you've survived the fast food industry. Sure, there are scars from numerous oil burns the grill gave for memory's sake and there are the few long-term co-workers you've ultimately become great friends with, but today's the big day. Today, you're turning in your two weeks' notice.
Three months later, sitting in class, you receive a text from Jacob. He's fed up with serving his flesh and blood for no worthwhile return, so that morning, he walked out. Jacob is the last of the co-workers you knew; not even Olivia works there anymore. Despite the horrors you endured while working there, your heart can't help but tug a little, but only a little, to return to fast food life. It's an ever-changing workplace, and you wanted to leave a memory. You should know better. You are nothing but a wave pounding on the sand.Endnotes
1. Scarily enough, at this particular fast food joint, most employees did not last even half a month before quitting or getting fired. Replacements were hired every week to maintain a stable number of workers.
2. Bet you didn't know that one of those measly hamburger meats costs the company a whopping $0.35. A single-meat burger costs outside customers $3.28, so yes, it really is a rip-off.
3. Mystery shoppers evaluate employees while disguised as customers. These evaluations are then submitted to the area manager, who determines which employees to fire. Some of the requirements for a drive-through worker to earn a perfect score include that he thank the customer for visiting, supply exactly two napkins per hamburger, and advertise the current special of the week.


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Search Results
Entries tagged with "
Marketing Funnel"
July 31, 201411:25 PM
The 7 Deadly Sins Of Digital Marketing In 2014 tagged "Marketing Funnel"
Have we solved the most basic of questions for our digital marketing efforts? I would argue that the vast majority of brands engaged in digital marketing have left the fundamentals behind them. These brands and are, at mass scale, following/chasing... Read more


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Artist description:
This is how they wanted themselves drawn. If you don't like how I drew you, It's your own fault for not being more specific in your descriptions.
Flash Draw Created May 24, 2007 470 x 338px Unless noted Copyright 2007 MonkeyNate.
We encourage you to submit a comment and let the artist know what you think of this drawing. Please log in or join to post a comment.
Post a comment
good pic, i love your style, just a lil' disproportionate, but not in to bad of a way, ~9
cool characters ! i like Rena
MAN that is one of the best drawings man keep it on u could do better than tat ,u ROCK!!
damn! *i still like your style :$
weeee i look cool :D is my new nickname angelface now? meh...great pic :D i style like your style XD
hope im not bothering ;((
lmao. awesome xD may i request jeans next time? (sorry to be fussy.) also, I have a trademark. I wear LOTS of bracelets (green&black beads).
Damn i look cool xD. You did did better than i was hoping :D Rena and angel look awsome too ^^
haha i love viirus :D he looks uber cool XD
Sorry Rena, I drew you too big and made your legs too short so they would fit in the picture.


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UFC 137 Fight Card: Nick Diaz Vacates Strikeforce Title to Fight at UFC 137June 9, 2011
Ahead of his October 27th match at UFC 137 with current UFC welterweight champion, Georges St-Pierre, Nick Diaz has vacated his Strikeforce welterweight title.
The news was broken by MMA Weekly, who says Strikeforce will crown a new champion.
According to MMA Weekly, UFC president Dana White said:
"He will give up the title, he will give up his Strikeforce title. He will come in and fight Georges St-Pierre, two guys will fight for the vacant belt in Strikeforce and we'll go from there."
Strikeforce is already lacking heavily in their welterweight department, and it is not yet known who could own the belt next.
The big names that come up from that shallow division are Tyron Woodley and Paul Daley. Daley recently lost to Diaz by first-round knockout in early April for the Strikeforce welterweight title.
There is also a bout with another one of the fighters who lost to Diaz in July containing Evangelista Santos who will face Tarec Saffiedine.
If Diaz so chooses, he could make a return to Strikeforce, but for now Diaz is strictly part of the UFC's welterweight division.
On October 27th, Diaz will catapult to the top of the UFC's welterweight division and fight GSP for the title in Las Vegas at Mandalay Bay.


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On Saturday, the final message of the General Synod of Bishops, entitled, 'The Eucharist: Living Bread for the Peace of the World', was made public in five languages by the Vatican.
It stresses, above all, a call to Eucharistic centrality in the life of all the faithful, and a particular call to priests and laity alike, to be "workers in the vineyard."
While many had speculated about dissent, particularly in the realm of celibacy and intercommunion, the bishops held closely to Church teaching and practice, stressing the importance of unity with the Magisterium and Chair of Peter--the Pope.
In the message, the Bishops say that they were, "Called to Rome by Pope John Paul II, of venerable memory, and confirmed by His Holiness Benedict XVI," to "come from the five continents of the world to pray and reflect together on the Eucharist, 'Source and Summit of the Life and Mission of the Church.'"
"The goal of the Synod" they point out, "was to offer proposals to the Holy Father that might help him to update and deepen the Eucharistic life of the Church. We have been able to experience what the Holy Eucharist has been from the very beginning: one faith and one Church, nourished by one bread of life, in visible communion with the successor of Peter."
The Bishops implored the faithful, "Christian brothers and sisters of every confession," to "pray more fervently that the day of reconciliation, and the full visible unity of the Church, might come in the celebration of the Holy Eucharist in conformity with Jesus' prayer on the eve of His death: 'That all may be one'."
They also recalled the testimonies, given by many bishops throughout the course of the Synod, on the various situations of relativism, secularism and even violent opposition in which the faithful live around the world.
In this light, the group vowed to continue to fight against abuses of human dignity and the scourge of relativism which seeks to undermine the Christian faith.
Liturgical reform?
Speaking specifically on the Eucharistic liturgy in the Mass, the Bishops declared that they "firmly believe and we teach in the constant tradition of the Church that the words of Jesus pronounced by the priest at the Mass, in the power of the Holy Spirit, effect what they signify. They bring about the real presence of the Risen Christ."
"Forty years after Vatican Council II", they said they "wanted to examine to what extent the mysteries of the faith are adequately expressed and celebrated in our liturgical assemblies."
The Synod reaffirmed "that Vatican Council II provided the necessary basis for an authentic liturgical renewal. It is necessary now to cultivate the positive fruits of this reform, and to correct abuses that have crept into liturgical practice."
"We are convinced", they said, "that respect for the sacred character of the liturgy is transmitted by genuine fidelity to liturgical norms of legitimate authority. No one should consider himself master of the Church's liturgy."
Here, the bishops noted many positive strides which have been made toward greater recognition and respect for the Eucharist in the Church.
These include; a "renewed consciousness of the importance of the Sunday Mass; the increase in the number of vocations to the priesthood and to consecrated life in various places of the world; the powerful experiences of World Youth Days, culminating at Cologne in Germany; the development of numerous initiatives for the adoration of the Blessed Sacrament almost everywhere in the world; the renewal of the catechesis on Baptism and the Eucharist in the light of the Catechism of the Catholic Church; the growth of movements and communities forming missionaries for the new evangelization."
At the same time however, they lamented a loss of the sense of sin and Sacrament of Penance among many of the faithful, as well as dropping Mass attendance and priest shortages in many regions.
Vocational challenge
Issuing a special call to priests and seminarians, the bishops challenged them to be, "with us and following the example of the Holy Father Pope Benedict XVI, 'humble workers in the vineyard of the Lord,' following a consistent priestly life."
"To all young seminarians," the bishops said that they wished "to express our hope that their formation will be permeated by an authentic Eucharistic spirituality."
Throughout the course of the Synod, the value of celibacy has been strongly reaffirmed as a valuable and vital practice, despite criticism from many referring to the sexual abuse scandal in the U.S.
The Bishop also extended their challenge of Eucharistic centrality to all married couples and lay people, as well as deacons and consecrated religious in particular.
The 257 prelates closed their message likening their own experience to that of the disciples whom Jesus appeared to on the road to Emmaus in the Gospels.
"At the end of this Synod", they wrote, "we experience that peace full of hope which the disciples of Emmaus, with burning hearts, received from the Risen Lord. They arose and returned in haste to Jerusalem, to share their joy with their brothers and sisters in the faith. We hope that you will go joyfully to meet Him in the Holy Eucharist, and that you will experience the truth of His words: 'And I am with you until the end of the world'."


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April 10, 2009
April 9, 2009
April 8, 2009
May 13, 2008
Michael New Jr. is the author of The San Francisco Giants Blog at http://www.michaelnewjr.com and is an avid fan. His favorite player is Will Clark. Go Giants!
Hey Michael—Jeff here, a Bleacher Report Community Coordinator.
I've been spending time on our Giants page and read a couple of your old articles. Really on-point analysis...you do good work.
Given that you haven't written in a while, I thought I'd check in and see if you're interested in picking up your coverage again. What do you say?
If not, it would be great if you could give me some feedback on why you decided to stop writing and what we could have done to better serve you. If you wouldn't mind, please shoot me a line at email@example.com with any thoughts.
Thanks so much,
Hey Michael,
Sorry for editing your article so many times... my internet is being strange and it won't let me leave you feedback. Anyway, I reworded a few things to make it clearer and added a new picture so check it out and see what you think.
Hey Michael,
I just wanted to let you know that Bleacher Report has launched out of beta with an updated look and feel and some great new features. Check out the "Game On" link in the header to find out more.
Next week we'll be launching the site to the world with some public announcements—and it'd be great to get a contribution from you between now and Tuesday to make Bleacher Report's launch a success.
Let me know if you need any help with ideas or have questions. Have a great weekend Michael!


text:
Build campaigns using InfusionSoft
Budget $250-750 USD
We have several initiatives and need a contractor to create multiple campaigns. In the future we may need those to be maintained as well.
4 freelancers are bidding on average $458 for this job
Hello Please be sure to use the chat feature here to discuss your project. I can help with Internet Marketing to drive sales. I have helped many others succeed and I am sure I can help you find success as well. I More
I have work experience of Affiliated Marketing and Affiliated Sales. I will look after Affiliated Sales Job with a pay for sale formula also. I have worked as a virtual assistant in the field of online customer ser More


text:
Artist description:
Bad thumb!
Flash Draw Created May 27, 2008 470 x 338px Unless noted Copyright 2008 crystallisa.
We encourage you to submit a comment and let the artist know what you think of this drawing. Please log in or join to post a comment.
Post a comment
It's amazing!
its so preety, I really liked the thumbnail! ^_^ 10!
So nice colors! 10!
thats really cool 10
Lovely draw 10++(:O)
Wow! Well drawn! Really Kool! ~!_ |[] _!~(Thx for comment)
wow this is so awsm 10
Can't have a new day if the stars didn't fade away
i like it , 10!
beautiful sky!


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A movie like Mary Kom is something that deserves the attention of even the president of India and we don't really know if that's what Priyanka Chopra visited the president for, but right before the release of the movie, she spent some time with Pranab Mukherjee and his wife at the Rashtrapathi Bhavan.
And looks like she loved the rendezvous with the first man of the country for this is what she said on her twitter about it, "Always incredible to be at the Rashtrapati Bhawan. A big thank you to the ever gracious Pranabda and Suvrama for having me at their home."
Priyanka Chopra was in the capital for a couple of days to promote Mary Kom. And just as the movie released today she flew off to Toronto to promote it their as it makes a premiere at the Toronto international film festival today. Here's wishing Priyanka Chopra good luck on the film's performance.d.getElementsByTagName('head')[0].appendChild(s);


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MGIMS Wardha
MGIMS Wardha Admission Notice has been issued by the regulatory authority for Admission in MBBS programme. Candidates who want to apply for the MBBS PMT 2016 they need to fill application form before the closing date.
|Organization Name
|MGIMS Wardha
|Closing date
|18th March 2016
|Exam Date
|17th April 2016
Selection will be on the base of merit list prepared after entrance exam held on 17th April 2016. From and prospectus of PMT admission 2016 are available till 11th march 2016.
The exam will consist of MCQ from subjects Physics, Chemistry and Biology. Recruitmentresult.com is publicizing rest of admission details for academic session 2016 are mentioned below.
MGIMS Wardha
MBBS PMT Application form 2016:
Eligibility criteria
- Candidate must have completed 17 years of age as on 31st December in the respective year of admission.
- Aspirants obtained a certificate of 12th standard under the system of Maharashtra State Higher Secondary educations with 50% marks or equivalent form a reputed university.
Medical Entrance Exams in India
Application form or applying procedure
The Prospectus and Application Form with syllabus, question papers of Gandhian thought for the last 6 years, with model question papers of Physics, Chemistry and Biology can be had from the office of the Dean either by:
- Paying a sum of Rs. 5000 (including exam fee) personally at the cash counter of Dean's Office
- Or by remitting a sum of Rs 5000/-(including exam fee) at any branch of State Bank of India by Challan which can be downloaded from the Institute website www.mgims.ac.in.
- The entrance exam will be held in Nagpur, Delhi, Hyderabad, Mumbai, Varanasi and Kolkata.
Also read- Scholarship In India
Prospectuses are available at office
The Dean,
Mahatma Gandhi Institute of Medical Sciences,
P.O. Sevagram,
Wardha – 442102
Important Dates
- Closing date: 18th March 2016
- Exam Date: 17th April 2016
Note: for further details must go through the Official Link
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List price: $17.99Publisher: St. Martin's PressPublication date: 5/30/2006Binding: PaperbackPages: 288Size: 5.75" wide x 8.25" long x 0.75" tall Weight: 0.594Language: English
Steven Saylor (born March 23, 1956) is an American author of historical novels. He is a graduate of the University of Texas at Austin, where he studied history and Classics. Although he also has written novels about Texas history, Saylor's best-known work is his Roma Sub Rosa series, set in ancient Rome. The novels' hero is a detective named Gordianus the Finder, active during the time of Sulla, Cicero, Julius Caesar, and Cleopatra. He divides his time residing in California and texas.
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FREMONT — It had been nearly 30 years since Elyria High made the hour-long trip down the turnpike to take on Fremont on the football field.
It may be another 30 before the Pioneers want to go back after the non-conference opener Friday.
The Pioneers had to come from 12 points down to salvage a 28-19 win over the Little Giants, a one-time conference rival.
"We're a young football team and we tried to keep our composure," said coach Kevin Fell, who won his fourth straight opener with Elyria. "It got to be a pretty physical game at the end and (Fremont) scared us to death. Every play, they could go. First game, you're going to get some goofy stuff."
Fremont had control early when it scored on back-to-back plays — kicking a 23-yard field goal and coming right back on the next play with an interception return by Jeffery Barnett to put the Pioneers in a 10-point hole with 5:05 left in the first quarter.
"We were down 19-7 and really hadn't done anything," said Fell, recalling Fremont's 68-yard scoring strike from quarterback Jalen Slick to Russ Yeast with 7:35 remaining in the third quarter. "Our quarterback (senior Hunter Parsh) finally settled down. We knew he would."
The Pioneers managed to keep the wheels on the wagon and eventually wore down Fremont with their defense and physical play.
"Elyria's a good football team," said Craig Yeast, Fremont's first-year coach. "They just continued to fight. We've got to learn to win. Right now, Elyria knows how to win."
Parsh finished with 304 passing yards, hitting senior Brad Schneider for a pair of scores and finding junior Kyle Ringer and senior Brendon Koziura with key strikes to keep possession. Ringer made two terrific catches in traffic on successive fourth downs to help Elyria run out the clock.
"Our line blocked for Hunter, and he just tried to get me the ball," Ringer said. "I just have to go and get it. It doesn't matter who's around me."
Schneider credited Elyria cornerback Mike Church, a three-year letterman.
"It helps when you go against the best DB in LorainCounty every day in practice," Schneider said. "And I love having Hunter as my QB. We've been together four years now."
Sophomore Chris Atkinson shook off an early fumble to help seal the win on the ground late. He finished with 84 yards on 23 carries behind Elyria's veteran line. Atkinson scored twice in the fourth quarter, the first giving Elyria its first lead and the second buttoning up the victory.
"We came out pretty slow," Parsh said. "We'll have to pick it up once we start playing some tough teams."
Elyria's defense was very good. Yes, it gave up the 68-yard pass play, but that was more of a great play by Yeast, the sophomore son of the coach. For the most part, Fremont struggled to sustain its offense.
Mike Smosny, the replacement for two-time All-Ohio linebacker Connor Kamczyk, tipped a Slick pass into the arms of Shamar Lucky to stop one Fremont drive and Orlando Scott covered a fumble to stop the Little Giants from scoring and possibly setting up an onside kick late.
"I knew if I caught it, it was a pick-six," Smosny said. "I tipped it, I looked back and (Lucky) caught it. We figured the defense was going to win us this game."
Elyria sported red helmets for the first time since the early 1960s. The Pioneers have worn white helmets with the red "E" on the side for almost the entire time since, the exception coming in Darrell Mayne's two-year reign in 1996-97 when Elyria wore black helmets.
Contact Tim Gebhardt at 329-7135 or email@example.com.


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Cottage Grove man sentenced for break-in, gun incident
A Cottage Grove man who broke into his parents' home and staged loaded guns and knives throughout the home to thwart an "alien invasion" will spend 10 years on probation.
Alexander C. Seaton, 20, was sentenced last month after pleading guilty in Washington County District Court to first-degree burglary with a deadly weapon. Two other charges were dismissed in a plea deal that stemmed from an incident nearly two years ago.
The criminal complaint describes the incident: Seaton, who was 18 at the time, broke into his parents' 85th Street Place home while they were out of town. He had been kicked out of the home three months prior.
Seaton sent a text message to his parents with a picture of himself holding two rifles. There were weapons in the home and not all had been locked away. Seaton's brother contacted him and the suspect admitted that he broke in through a basement window. He sounded "extremely paranoid and was talking about government conspiracies and people following him," according to the complaint.
Police and the Washington County Special Response Team went to the home and tried to contact Seaton by phone. Neighbors reported hearing gunshots from the residence.
Later, Seaton walked out of the home holding a cellphone and appeared startled by the police presence. He was safely apprehended.
Inside the home police found numerous loaded guns and unsheathed knives placed by doors and windows. A safe was damaged and discharged shotgun shells were found outside the front and back doors.
In an interview with an investigator, an irritated Seaton said he had stayed in his parents' garage for two days before "the voices (in his head) told him to break into the house," the complaint says. He admitted to shooting five rounds outside the home and said he was told by voices to break into the home and load weapons to "stave off an alien invasion."
The court issued a stay of imposition on his sentence for the felony crime and placed him on 10 years probation with numerous conditions, including that he pay a fine, remain law-abiding, not use drugs or alcohol, complete an offender recovery program and finish all mental health programming.


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Bipartisan bill would spur innovation, competitiveness, say authors
U.S. Senators Amy Klobuchar (D-MN) and Scott Brown (R-MA) introduced bipartisan legislation this week that they say would help revitalize America's innovative edge and ability to compete in the global economy.
The Innovate America Act would cut red tape to help businesses utilize research and development for new products, target successful education programs, and promote U.S. exports in new markets to strengthen America's ability to innovate and compete in the global economy.
"Innovation has always been a catalyzing force in Minnesota's economy," Klobuchar said.
"By cutting red tape for businesses and focusing on science, technology, engineering, and math education, we can help our businesses attract and retain our country's brightest scientists, engineers, and researchers. This bill shows we can come together on a competitive agenda that will move America forward."
"The Innovate America Act will create jobs and foster innovation by establishing a more favorable environment for small businesses and entrepreneurs everywhere," Brown said. "During these challenging economic times, it is imperative that we give our universities in Massachusetts and throughout the country the tools and resources they need to turn their ideas into companies and jobs. We also must ensure that our manufacturers and exporters are able to compete in the global marketplace."
Klobuchar serves on the Senate Commerce Committee; chairs the Subcommittee on Innovation, Competitiveness, and Export Promotion; and recently held an "Innovation Summit" attended by hundreds of business leaders and policy experts to discuss public and private strategies to strengthen America's ability to compete globally.
Klobuchar traveled to more 20 Minnesota cities, including Alexandria, in January highlighting the potential for economic innovation to drive long-term prosperity and job creation.
Brown was recently named to the Senate Committee on Small Business and Entrepreneurship in the 112th Congress. Last year, he received the Manufacturing Legislative Excellence Award from the National Association of Manufacturers.
The legislation would also help increase the competitiveness of small- and medium-sized businesses by focusing on turning research and new technologies created at universities into products; promoting and rewarding schools, technical colleges, and universities that focus on science, technology, engineering and math; and removing red tape and reducing production costs for manufacturing businesses.
Klobuchar added, "In recent years, the country has fallen behind in its efforts to research, develop, and compete in the global economy. We are resting on our laurels at a time when other countries, including China and India, are moving full-steam ahead. This legislation can help create a new wave of invention, entrepreneurship, and exports in the short term, while laying the education foundation to secure America's standing in the global economy in the long term."
"The challenges we face as a nation require all of us working together to get our economy moving again, and I am proud of this bipartisan legislation that will spur American innovation and entrepreneurship," Brown concluded.
Specifically the legislation would:
--Encourage greater commercialization of Research and Development by expanding the Basic Research Tax Credit to include all industry-funded university research
--Allow companies to take a flat 30-percent tax credit for donating equipment to high schools and technical and community colleges
--Fund 100 new Science, Technology, Engineering, and Mathematics (STEM)-focused high schools
--Provide incentives to colleges and universities to increase graduation rates for STEM students
--Expand undergraduate research opportunities
--Establish a Website for technology commercialization ideas
--Establish a manufacturing assistance program
--Remove regulatory barriers for top 20 exporting industries
--Enforce existing international exporting and importing laws
--Reduce unnecessary printing and publishing costs of government documents
--Eliminate bonuses for poor performance by government contractors


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Lyres (The) - Some Lyres (Music CD)
Release Date: 08 November 2010
Artist: Lyres (The)
£5.89 includes FREE UK Postage (other delivery options available)
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Falkland Islands
An excellent, if brief (12 tracks) part-career retrospective, part-collection of oddities that includes their 1979 debut single "How Do You Know" b/w "Don't Give it Up Now." Packaged as a parody of the Rolling Stones' controversial Some Girls, Some Lyres proves to be a valuable introduction. ~ John Dougan
Release Date: 08 November 2010
Artist: Lyres (The)
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I'm probably late to the game, but I didn't realize until last night that PHP4 is included by default in Mac OS X (10.1). All you need to do is activate it. With a little snooping around I found some simple instructions on how to do this here (scroll down about 3/4 of the way and see the command line steps under "Activating the PHP Module"). In 2 minutes I was building a site with PHP on my local machine.
Salon: "'You've probably been thanked for 'Ferris Bueller' a thousand times, so I'll just thank you for a movie you weren't even in, as a sort of acknowledged irony between two clever chaps.' But actually, it was more like, 'Hi Matthew, I'm stupid — no, just kidding, but I am tremendously ludicrous and abstruse. And did you know that I like to pee in my own socks?'" — What happens when you try to be funny while meeting Matthew Broderick in line at the movie theatre.


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The St. Paul school board has extended the contract of Superintendent Valeria Silva for three years. Her new contract will take effect when her current deal expires in December.
With a 2 percent raise and a possible 1 percent performance bonus, she'll make about $193,000 a year.
Silva can get similar pay hikes in following years, along with an $11,000 annual longevity payment for having worked for the district for more than two decades.
School Board Chairwoman Jean O'Connell praised Silva's performance.
"We feel that she's done really solid work in communications, strategic planning, leadership of the team, instructional leadership. And we expect even better results as we go forward," O'Connell said.
Silva's commitment to the district is very valuable, O'Connell said.
"To have that stability in the superintendent's position is absolutely key to keeping the organization moving forward," O'Connell said. "In effect, we have set ourselves up to have a superintendent at least for six [years] and she is interested in even longer tenure."
Silva has been the district's superintendent since late 2009. And she's been an employee for 25 years, starting as an educational assistant in the district's Spanish language immersion program.


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Charlie Carter looked at the blackened, bare framework of what used to be his bus and wept.
Firefighters were hosing down the smoking ruin of the Ford Econoline that Carter has used for the past five years to carry his six severely disabled foster children.
''I don't know how we're going to replace it,'' he said. ''We've got a lot of memories in it.''
Carter will remember the trips to Cherokee, N.C., with his ''kids'' -- wheelchair-bound foster children ranging in age from 8 to 26. He'll remember carrying a little girl named Whitney to see the beach in Jacksonville, Fla. The 8-year-old, on a respirator and paralyzed from the neck down, died on the return trip, her wish fulfilled.
Whitney christened the lift-equipped bus with the name ''Charlie's Angels,'' a moniker that was airbrushed on the side.
''We lost a child on that bus and everything,'' Carter said, his voice choked with tears.
The fire that engulfed the bus in the yard of Carter's Hughes Street home Wednesday afternoon moved quickly. Carter was readying the bus for a trip to have 20-year-old Joel Grier fitted for a new wheelchair when he spotted flames spitting from the dashboard. He suspects an electrical short was the cause.
''It went fast,'' he said. ''I just filled it up with gas the other day.''
An insurance policy may cover the $3,500 Carter and his wife, Kaye, paid for the bus at a county auction. But most handicapped-equipped buses, even used, cost at least three times that, Carter said. Without it, the Carters can't take more than two of their foster children at a time fishing or to the doctor's office.
''There ain't no way I can replace it, at that price,'' said Carter, who makes do with a disability check and the $500 a month the children get. ''I don't know what we're going to do.''
Anyone interested in offering assistance can call Charles Carter at (706) 549-2928.
Courts reporter Stephen Gurr can be reached at email@example.com or (706) 208-2219.
This article published in the Athens Daily News on Thursday, April 19, 2001.
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The situation was eerily similar.
After grabbing the rebound off a missed free throw in a game against West Virginia, the final seconds ticking away, Chris Walker called a timeout.
Rewind to Feb. 16. Tech was pushing the ball up the court with about 10 seconds left when Walker called time, the Red Raiders trailing by two points. It appeared as if Tech may have had numbers, but Walker said he didn't want to see the game end with his team turning the ball over, having been unable to get off a final shot.
But all Tech could muster in the final possession was a contested Josh Gray 3-pointer that glanced harmlessly off the rim.
The only difference Wednesday night was that the game was tied. Walker stuck to his guns and called for a time out. This time the staff made it clear they wanted a play going to the basket, unless they could free up sharpshooter Dusty Hannahs for a 3-pointer.
Hannahs couldn't shake loose, so Gray went to work. He attacked the rim. His shot got blocked, but as is often the case, action going toward the basket resulted in a second opportunity. The block went right to Jamal Williams. His free was off the mark, but with West Virginia inexplicably failing to box out, Kravic was all alone for the game-winning put back.
The whole sequence illsutrated that Tech had learned from the lessons of last month's sour defeat in Morgantown. It showed growth by a young head coach and a new level of maturity for an inexperienced team that is playing perhaps its best basketball of the season.


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food packaging, plastic container, disposable container, disposable cup, plastic packaging, Manufacturer and Supplier - Taiwan
As a leading food packaging manufacturer, Jadebox Packaging is dedicated in manufacturing & worldwide export. We specialized in food service packaging for supermarket and food processing industry. We have extensive range of thermoform plastics packagings: Blisters, Clamshells, Trays, Containers, Box... More information


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Farm Dreams workshop scheduled Jan. 8 in Clinton
Is Farming in Your Future?
Land Stewardship Project's "Farm Dreams" Workshop gives participants the opportunity to explore this question with an upcoming workshop:
Sunday, Jan. 8 from 1-5 p.m. in Clinton, Minn.
The registration fee is $20 for Land Stewardship Project (LSP) members and $40 for non-members. Pre-registration is required. For more details or to register, contact: Nick Olson at 320-269-2105 or email@example.com
This workshop is designed to help people who are seeking practical, common sense information on whether farming is the next step for them. In this workshop you will assess your resources, skills and motivations for farming; plan your educational path toward farming; learn about regional training opportunities and support networks and prioritize your next steps.
Land Stewardship Project is a grassroots membership organization made up of farmers, as well as rural and urban residents working together for a food an agriculture system that makes sense for the land and for the people. To learn more and to join, visit: www.landstewardshipproject.org
The "Farm Dreams" initiative is supported by the Beginning Farmer and Rancher Development Program of the National Institute of Food and Agriculture, USDA, Grant 2010-03107.