Correspondence: Shu-Wen Chang, Department of Ophthalmology, Far Eastern Memorial Hospital, No. 21, Section 2, Nanya S. Road, New Taipei City 220, Taiwan, Republic of China; swchang2007@ntu.edu.tw.
Tsan-Chi Chen, Shu-Wen Chang; Moxifloxacin Modulated TGF-β1-Related Interleukin-12 Secretion in Corneal Fibroblasts. Invest. Ophthalmol. Vis. Sci. 2017;58(13):5692-5702. doi: 10.1167/iovs.17-21944.
Purpose: To explore the variability in interleukin-12 (IL-12) secretion in human corneal fibroblasts (HCFs), the effects of IL-12 on HCF viability and migration, and the effects of recombinant human transforming growth factor-β1 (rhTGF-β1) and moxifloxacin (MOX) on IL-12 expression.
Methods: IL-12 secretion in 27 primary HCFs incubated with rhTGF-β1 and/or MOX was analyzed using immunoblotting. Viability and migration were assessed using WST-1 assay and culture inserts, respectively. The effects of IL-12 on HCFs were verified via adding recombinant human IL-12 (rhIL-12) or silencing IL-12. The effects of rhTGF-β1 and MOX on IL-12 secretion were also verified.
Results: There were 25.9% HCFs that did not secrete IL-12 and 74.1% HCFs that secreted IL-12 were divided into low- (25.9%), medium- (37.0%), and high-secretion (11.1%) subgroups. After 3-day incubation, rhTGF-β1 stimulated IL-12 by 1.90 ± 0.84 folds (P = 0.026), with no difference in folds of induction among subgroups (P = 0.870). MOX suppressed IL-12 secretion concentration-dependently, more noticeably in the presence of rhTGF-β1. Exogenous rhIL-12 enhanced HCF proliferation (1.29 folds, P < 0.001) and migration. In contrast, rhTGF-β1 only enhanced proliferation (1.24 folds, P < 0.001). In IL-12-silenced HCFs, rhTGF-β1 failed to enhance IL-12 secretion and resulted in reduced cell proliferation and migration.
Conclusions: IL-12 enhanced HCF proliferation and migration. TGF-β1 per se enhanced HCF proliferation but not migration. However, it promoted migration indirectly through upregulating IL-12 secretion. MOX-inhibited HCF migration via suppressing IL-12 secretion. The interindividual variation in IL-12 secretion might contribute to disparity in HCF proliferation and migration after corneal wounding among individuals.
Regulation of corneal stromal cells in corneal wound healing is mediated in part by growth factors,1–3 cytokines,4 and extracellular components.5 Corneal keratocytes are normally quiescent and could differentiate into fibroblasts and myofibroblasts in response to corneal epithelium-derived growth factors such as platelet-derived growth factor and transforming growth factor-β1 (TGF-β1).5–7 In addition, exposure to TGF-β1 increases TGF-β1 secretion through an autocrine process in human corneal fibroblasts (HCFs).8 It is thus inspiring to illustrate whether the increased autocrine TGF-β1 in cornea stromal cells further induce other intracellular responses in themselves.
Interleukin-12 (IL-12) is a covalently linked heterodimer with a light chain IL-12A (p35) and a heavy chain IL-12B (p40), which controls its biological activity.9 IL-12 is a proinflammatory cytokine that activates natural killer cells and T lymphocytes.10 It also inhibits apoptosis of T cells.11 However, the production, function, and regulation of IL-12 in noninflammatory cells remain less well understood.
In the eye, IL-12 is present in the tear at 89.8 ± 115.8 pg/mL.12 Its expression increased significantly in the corneas with fungal13 and herpetic stromal keratitis.14 It also increased in the tears of patients following refractive surgery.15 In vitro study on the stimulation of secretory IL-12 in noninflammatory cells is limited. There is only one study illustrating expression of IL-35, which contains IL-12A as a member of the IL-12 family, following TGF-β1 induction in dermal fibroblasts.16 In contrast, interferon-β suppresses the production of IL-12B production in dendritic cells.17 Whether corneal fibroblasts participate in the regulation of IL-12 and/or if there is an interindividual variation in IL-12 secretion remain less well explored.
Topical moxifloxacin (MOX), a four-generation fluoroquinolone, is frequently used prophylactically after refractive surgery,18,19 cataract surgery,20,21 and intravitreous injection.22,23 In addition to its well-known antimicrobial activity, MOX also modifies corneal fibroblast-to-myofibroblast differentiation in HCFs.8 Furthermore, MOX suppresses inflammatory responses by reducing IL-1β and IL-17A in pneumonia mice.24 Therefore, it is inspiring whether MOX regulates the IL-12 secretion in corneal stromal cells. As reducing inflammation is beneficial in corneal infection as well as following surgery, we investigated if corneal fibroblasts produce proinflammatory cytokine IL-12 and its effects on corneal stromal cells proliferation and migration. We also investigated the interindividual variation of IL-12 and its alteration after MOX application.
This study was approved by the Institutional Research Ethics Review Committee (Protocol No. FEMH-100102-F). Human corneal rims were harvested as the source of corneal fibroblasts after the central corneas were removed for transplantation. A total of 27 donors aged between 20 and 70 years were included and all met the inclusion and exclusion criteria according to the Eye Bank Association of America standards for human corneal transplantation.25 In the laboratory, the residual corneal rims were digested in collagenase-containing (2 mg/mL; Roche Applied Science, Mannheim, Germany) Dulbecco's modified Eagle's medium (DMEM; Gibco, Grand Island, NY, USA) with 10% heat-inactive fetal bovine serum (FBS; Biological Industries, Kibbutz Beit Haemek, Israel), 200 unit/mL penicillin G, and 200 μg/mL streptomycin sulfate (Sigma-Aldrich Corp., St. Louis, MO, USA) for 6 to 8 hours. Then, the digested cells were resuspended and maintained in DMEM with 10% heat-inactive FBS, 100 unit/mL penicillin G, and 100 μg/mL streptomycin sulfate as a complete culture medium under a humidified atmosphere containing 5% CO2 at 37°C. The isolated HCFs were used in passages 4 to 10 with complete culture medium, containing 10% FBS.
To reduce IL-12 secretion, the lentiviral short hairpin RNAi (shRNA) construct shIL12B obtained from the National RNAi Core Facility in Academia Sinica, Taiwan, were used to infect HCFs as the procedures with minor modification.1 Briefly, HCFs were infected continuously for 3 days with lentivirus-based shIL12B pseudovirion at 10 multiplicity of infection, which was produced from pseudovirion-packaging 293T cells with the clone pLKO.1-shIL12B (TRCN0000058760, 5′-GTCGTAGAATTGGATTGGTAT-3′) using X-treme HP Transfection Reagent (Roche, Indianapolis, IN, USA).
To analyze the effect of TGF-β1 and MOX on IL-12 secretion, 2.0 × 105 cells were seeded into a 6-cm culture dish and treated on the next day with 3 mL serum-less culture medium containing 0.1% FBS/DMEM, 2 ng/mL recombinant human TGF-β1 (rhTGF-β1; R&D Systems, Minneapolis, MN, USA), and/or MOX (VIGAMOX; Alcon Laboratory, Inc., Fort Worth, TX, USA) at 0, 10, 50, or 100 μg/mL.
To analyze the effects of conditioned media on cell proliferation and migration, HCFs with/without IL-12 silencing were cultured in a 6-cm culture dish as described above. The conditioned media were collected at 3 days post incubation (dpi) of TGF-β1 and/or MOX 100 μg/mL, respectively. In order to adjust the equal amount of the conditioned medium, the adhesion cells were harvested and disrupted in lysis buffers (150 mM NaCl, 0.5% sodium deoxycholate, 1% NP-40, 50 mM Tris-HCl, pH 7.4) for total protein determination using a bicinchoninic acid (BCA) Protein Assay kit (Pierce, Rockford, IL, USA). The ratio of total protein amounts of cell lysates between control group and individual treatment group was calculated separately for each dish. Subsequently, appropriate amount of DMEM was added to the conditioned media according to the inverse of the ratio to form the adjusted conditioned medium (ACM) for the subsequent experiments. The ACM was considered standardized and contained equal amount of cytokines and growth factors per contributing cell.
To evaluate the effect of exogenous IL-12 on HCF proliferation, HCFs were cultivated in 100 μL serum-less DMEM containing recombinant human IL-12 (rhIL-12; ANTEC Bioscience, Inc., Taipei, Taiwan) and/or MOX for 3 days. In addition, to evaluate the effect of conditioned medium on HCFs, 100 μL ACM collected from the nonsilenced or IL-12-silenced HCFs were used to culture HCFs. HCF cell growth was evaluated by the formazan-based Cell Proliferation Reagent WST-1 assay (Roche Applied Science) according to manufacturer's instructions. Briefly, 5.0 × 103 cells were cultivated individually in each well in a 96-well plate, and the incubation medium was refreshed on the next day according to the above-mentioned conditions. For the above assay conditions, 10 μL WST-1 reagent were directly added into each well at the detection times and subsequently incubated at 37°C for 2 hours. Absorbance of the samples was measured at 450 nm against a reference at 690 nm using a μQuant microplate spectrophotometer with KC Junior software (BioTek Instruments, Inc., Winooski, VT, USA).
To compare the effect of IL-12, TGF-β1, or conditioned medium on the motility of HCFs, each area of Culture Inserts (ibidi GmbH, Martinsried, Germany) was seeded with 1.0 × 104 cells in complete DMEM and cultured for 1 day in a six-well culture dish. The area surrounding the Culture Inserts remained cell-free. After removing the Culture Inserts, the cells were further cultured in 2 mL serum-less DMEM containing 10 ng/ml rhIL-12, 10 ng/mL rhTGF-β1, or ACM of the treated HCFs. The HCFs were allowed to migrate, and the images were directly captured every 1 hour for 3 days using a Real-Time Cultured Monitoring System with Multiple-Point Imaging Capture (CCM-MULTI; ASTEC Co., Ltd., Fukuoka, Japan). The number of migratory cells was counted and the difference among the conditions analyzed.
To analyze the expression of secretory IL-12 in HCFs, 10 μL of each ACM sample was mixed in 5 μL 3× sample buffer (180 mM Tris-HCl, pH 6.8, 30% glycerol, 0.1% bromophenol blue) without β-mercaptoethanol and then analyzed by 10% SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA). Following blocking nonspecific binding with 5% bovine serum albumin (BSA) in Tris-buffered saline with Tween (TBST; 20 mM Tris-HCl, pH 7.5, 0.5 M NaCl, and 0.1% Tween-20) for 1 hour, the membranes were incubated at 4°C overnight in 5% BSA/TBST with the primary IL-12B-specific antibody (#5924-1; 1:1000; rabbit monoclonal; Epitomics, Inc., Burlingame, CA, USA). The membranes were subsequently incubated at room temperature with horseradish peroxidase (HRP)-conjugated antirabbit secondary antibodies for 1 hour. Finally, the blots were developed by chemiluminescence (Millipore) and analyzed by FUJIFILM LAS-4000 Imaging System (FUJIFILM, Tokyo, Japan).
To estimate the secretory IL-12 concentration, a twofold serial dilutions of rhIL-12 ranging from 0.625 to 20 ng/mL were mixed with 3× sample buffer in a final volume of 15 μL and analyzed by immunoblot for each experiment. The density of the blots was digitized and an equation of the IL-12 standard curve by linear regression was generated for every experiment. All the equivalent secretory IL-12 concentrations of the individual experiment conditions were then calculated accordingly.
To determine the silence efficiency, total RNA in nonsilenced and IL-12-silenced HCFs was extracted by RNeasy Mini Extraction kit (QIAGEN, Valencia, CA, USA) according to the manufacturer's recommendations. The first-strand cDNA was synthesized with the reaction containing all extract of total RNA in a total volume of 30 μL containing 20 pmol of oligo(dT)15, 10 pmol of dNTP mix, and 200 units of MMLV Reverse Transcriptase (Promega, Madison, MI, USA). A fluorescein quantitative PCR detection system (Clontech, Mountain View, CA, USA) was used to evaluate the expressions in transcription levels using the specific primer sets, namely 5′-TCAGTGTCAAAAGCAGCAGA-3′ and 5′-TGATGATGTCCCTGATGAAG-3′ for IL-12; 5′-CACCACCAACTGCTTAG-3′ and 5′-CTTCACCACCTTCTTGATG-3′ for GAPDH; 5′-TCCTGTGGCATCCACGAA-3′ and 5′-ATGATCTTGATCTTCATT-3′ for β-actin; 5′-CAGAAGGTGACCAGCCTAAC and 5′-GTTCTGGGAGGAGATGGGTG-3′ for Thy-1; 5′-CCCACCAGAGGAGTATGTCC-3′ and 5′-CCCAGGTCAGAGTTCAATAG-3′ for FAK. The amount of IL-12 mRNA was normalized to mRNA of GAPDH, β-actin, Thy-1, and FAK, respectively. The fluorescence results were analyzed with the LightCycler Software version 3.5 (Roche Applied Science).
To determine the concentration of TGF-β1 in the media, 100 μL ACM from the treated HCFs in a 6-cm dish was loaded on PVDF membrane (Millipore). After blocking nonspecific binding with 5% BSA (Sigma-Aldrich Corp.) in TBST for 1 hour, membranes were incubated at 4°C overnight in 5% BSA containing TBST with the TGF-β1-specific primary antibody (#5249; 1:1000, rabbit monoclonal; Epitomics, Inc.). The PVDF membranes were then incubated at room temperature with HRP-conjugated antirabbit (1:10,000) secondary antibody for 1 hour. Finally, the blots were developed with ECL chemiluminescence (Millipore) and analyzed with a FUJIFILM LAS-4000 Imaging System (FUJIFILM). The density of the blots was digitized and folds of TGF-β1 concentration change estimated.
Each triplicate experiment was performed independently for at least three times as described in figure legends. The means of each triplicates of the indicated parameters were used for further analysis to investigate whether there was difference among different groups in each experiment. The differences in IL-12 concentration, folds of IL-12 concentration change, cell viability, and number of migratory cells among groups were evaluated using 1-way ANOVA followed by Tukey's honestly significant difference (HSD) post hoc test according to the experimental design as described in figure legends. The difference in donors' age between the secretory and nonsecretory groups was examined using Student's t-test. The correlation between the folds of increase in TGF-β1 and IL-12 concentration in the conditioned media was examined using Pearson's two-tailed correlation. A P value less than 0.05 was considered statistically significant. All the statistics were conducted by SPSS version 16.0 software (SPSS, Inc., Chicago, IL, USA).
The secretory IL-12 concentrations in the media were estimated using an IL-12 equivalent equation (Fig. 1A). The 27 isolated HCFs were first classified as nonsecretory (n = 7) and secretory groups (n = 20) according to the signal intensity of the baseline IL-12 on immunoblots (Fig. 1B). The secretory group was further subdivided into three subgroups according to the estimated baseline IL-12 concentrations on immunoblots; namely low secretory group, medium secretory group, and high secretory group. The donors averaged 55.6 ± 18.6 and 56.5 ± 16.5 years old in the IL-12 secretory and nonsecretory group, respectively. There was no significant difference between these two groups (P = 0.92). To mimic the IL-12 expression during corneal wound healing, exogenous rhTGF-β1 was added to HCFs. Two of the nonsecretory HCFs responded to rhTGF-β1 to secrete IL-12, while five of them remained nonsecretory. One representative rhTGF-β1 stimulated IL-12 response in each secretory HCFs group is shown in Figure 1C. There was more IL-12 secretion in the high secretory group. However, there was no significant difference in the folds of TGF-β1-stimulated secretory IL-12 among the three subgroups (P = 0.870), which averaged to 1.90 ± 0.84 folds. We also found a 1.54 ± 0.67-fold increase in TGF-β1 concentration in the conditioned media of TGF-β1 treated cells at 3 days. The folds of increase in TGF-β1 concentration correlated significantly with the folds of increase in IL-12 concentration (correlation coefficient r = 0.454, P = 0.044).
Classification of the isolated HCFs regarding IL-12 secretion. (A) One example of twofold serial dilutions of rhIL-12 in serum-less, ranging from 0.625 to 20 ng/mL, were determined by immunoblot with IL-12–specific antibody to generate a standard curve equation. For this blot, y = 0.1345x + 0.8634 (y value is the concentration of secretory IL-12 and x value is the signal intensity of the band; R2 = 0.9638). (B) According to the signal intensity of the baseline IL-12 secretion on immunoblots, the 27 isolated HCFs were divided into nonsecretory and secretory groups. The secretory group were then further divided into three subgroups according to the calculated baseline concentration of secretory IL-12 in HCFs. (C) One representative conditioned media from the three subgroups were analyzed at 3 dpi of 2 ng/mL rhTGF-β1. The concentration of secretory IL-12 was calculated separately and shown as the numbers below the blots.
To illustrate the temporal IL-12 expression during corneal wound healing, exogenous rhTGF-β1 was added to three of the secretory HCFs and the secreted IL-12 was detected by immunoblotting. We illustrated that continuous rhTGF-β1 incubation significantly induced IL-12 secretion in a time-dependent manner; that is, 5.02 ± 0.53 folds at 2 dpi (P < 0.001) and 6.92 ± 0.87 folds at 3 dpi (P < 0.001) (Fig. 2).
TGF-β1 upregulated IL-12 secretion in HCFs. Cells were incubated in serum-less medium with rhTGF-β1 at 2 ng/mL, and the conditioned media were harvested at the indicated incubation time points. Secretory IL-12 in conditioned media was analyzed by immunoblotting with IL-12–specific antibody and the folds of IL-12 induction calculated. The quantitative data in the histograms represent the mean ± SD of the individual experiments performed in different cell sources (n = 3). Differences in the histograms were analyzed by the 1-way ANOVA followed by Tukey's HSD. Numbers represent P values between the two indicated time periods. u.d., undetectable.
In the absence of rhTGF-β1, MOX inhibited IL-12 secretion concentration-dependently (Fig. 3, lanes 1–4, ANOVA, P = 0.001). The difference was statistically significant at 50 and 100 μg/mL (lanes 3 and 4 versus lane 1, P = 0.007 and < 0.001, respectively). While rhTGF-β1 significantly induced IL-12 secretion by approximate 1.67 folds (Fig. 3, lane 5 versus lane 1, P < 0.001), the increase was also diminished by MOX concentration-dependently (Fig. 3, lanes 5–8, ANOVA, P < 0.001). The difference was significant at concentration of higher than 10 μg/mL and concentration-dependently.
MOX suppressed IL-12 secretion in HCFs. Cells were incubated in serum-less medium with rhTGF-β1 at 2 ng/mL and/or MOX at the indicated concentration for 3 days. The conditioned media were harvested at 3 dpi. The secretory IL-12 was analyzed by immunoblotting with IL-12–specific antibody and the folds of IL-12 induction calculated. The quantitative data in the histograms represent the mean ± SD of the individual experiments performed in different cell sources (n = 5). Differences in the histograms were analyzed by 1-way ANOVA followed by Tukey's HSD. Numbers represent P values between the two indicated groups.
To verify the effect of secretory IL-12 on corneal stromal cell proliferation, rhIL-12 at 0.1, 1, and 10 ng/mL were added to incubate HCFs in serum-less culture medium for 3 days. We illustrated that rhIL-12 incubation significantly enhanced HCFs proliferation (Fig. 4A), suggesting that IL-12 promoted corneal stromal cell proliferation. The difference for 1 ng/mL IL-12 was significant since day 2, while the difference for 10 ng/ml IL-12 was significant since day 1. We also illustrated that MOX incubation significantly diminished the rhIL-12 enhanced HCF proliferation (Fig. 4B).
MOX suppressed IL-12-enhanced HCF cell growth. (A) rhIL-12 at 0.1, 1, and 10 ng/mL were added to the culture media, and viability of HCFs was determined at days 0, 1, 2, and 3. (B) MOX at 100 μg/mL and rhIL-12 at 10 ng/mL were added to the culture media. The HCF viability was determined and calibrated to the individual viability on day 0 at the indicated time points. The quantitative data represent mean ± SD of the individual experiments performed in different cell sources (n = 3). Differences in the relative changes in cell viability were analyzed by 1-way ANOVA followed by Tukey's HSD. Numbers represent P values between the two indicated groups.
We also investigated whether secretory IL-12 affects HCF migration. Using a migration assay, HCFs were added to the center of an ibidi culture inserts and incubated in 6-cm culture dishes. Either 10 ng/mL rhIL-12 or 10 ng/mL rhTGF-β1 was added to the serum-less medium (Fig. 5). The results revealed that rhIL-12, but not rhTGF-β1, enhanced HCF migration at dpi 3 (Fig. 5C, 141.8 ± 32.9 vs. 93.3 ± 18.4, P = 0.034 and 65.0 ± 19.7 vs. 93.3 ± 18.4, P = 0.520, for rhIL-12 and rhTGF-β1, respectively). These suggest that increased IL-12 secretion in HCF could promote HCF migration.
Enhancement of IL-12 on HCF migration. (A) A scheme depicting the location of image capture in a culture insert in a six-well culture dish and the area surrounding culture inserts remained cell-free. After removing the culture insert, 2 mL serum-less DMEM without (as control) or with rhIL-12 or TGF-β1 at 10 ng/mL were added into the dish to incubate the HCFs for 3 days. (B) Cell migration of IL-12– or TGF-β1–incubated HCFs was monitored daily, and the images captured at 3 dpi were shown. HCF migration was performed in different cell sources (n = 3). (C) The number of migratory HCFs that passed the dotted line, which indicates original ibidi insert margin, was counted. The quantitative data represent mean ± SD of three individual experiments performed in different cell sources with similar trend. The difference among the three groups were analyzed by 1-way ANOVA followed by Tukey's HSD. Numbers represent P values between the two indicated groups.
Using the gene silence method, we had generated shIL12 pseudovirions efficiently to decrease the IL-12 expression in HCFs by approximately 80% at transcription level (Fig. 6A). The HCFs, with or without IL-12 silencing, were treated with MOX and/or rhTGF-β1 in a serum-less medium (0.1% FBS) for 3 days. The secretory IL-12 and TGF-β1 in the conditioned medium were determined by immunoblotting (Fig. 6B) and dot blotting (Fig. 6C), respectively. TGF-β1 significantly enhanced IL-12 secretion while MOX significantly inhibited TGF-β1 related IL-12 secretion (Fig. 6B, lanes 1–4). There was almost no detectable secretory IL-12 in the conditioned medium of IL-12- silenced HCFs (Fig. 6B, lanes 5–8). Addition of TGF-β1 resulted in significantly higher TGF-β1 concentration in the conditioned media (Fig. 6C, dot 3 vs. dot 9). In addition, there was similar TGF-β1 concentration in the medium between nonsilenced and silenced HCFs (Fig. 6C, dot 3 vs. dot 7, P = 0.656). This indicates that silencing IL-12 did not interfere with the auto-regulation of TGF-β1 secretion in HCFs.
Secretory IL-12 and TGF-β1 expression in the IL-12-silenced HCFs. (A) IL-12 gene in HCFs was reduced by infection system with the lentivirus-based shIL12 pseudovirion and cultured in complete culture medium with 10% FBS. Silence efficiency on IL-12 expression was analyzed by semiquantitative real-time PCR with IL-12-specific primers. Differences in the relative changes in silence efficiency were analyzed by the Student's t-test. (B) Conditioned media of nonsilenced or IL-12-silenced HCFs were harvested after 3-day incubation in the serum-less DMEM with 2 ng/mL rhTGF-β1 and/or 100 μg/mL MOX. The amount of IL-12 secretion in each ACM were determined by immunoblotting with IL-12-specific antibody and the folds of IL-12 induction calculated. Differences among groups were analyzed by 1-way ANOVA followed by Tukey's HSD. (C) The amount of TGF-β1 in the same ACM were determined by dot blotting with TGF-β1-specific antibody. The standard indicates 2 ng/mL TGF-β1 used for HCF culture. The quantitative data in the histograms represent mean ± SD of three individual experiments performed in different cell source (n = 3). Differences among groups were analyzed by 1-way ANOVA followed by Tukey's HSD. Numbers represent P values between the two indicated groups.
The above-mentioned conditioned media were further used to incubate HCFs to verify their effects on cell motility and cell proliferation (Fig. 7). The conditioned medium from the nonsilenced, rhTGF-β1-treated HCFs significantly enhanced motility (Figs. 7B, 7C, panel c) and cell proliferation (Fig. 8, 1.91 ± 0.06 vs. 1.54 ± 0.04 folds at 3 days, P < 0.001). In contrast, the conditioned medium from IL-12-silenced HCFs completely retarded HCF migration (Figs. 7B, 7C, panels e–h) but did not suppress HCF proliferation (Fig. 8, 1.36 ± 0.08 vs. 1.54 ± 0.04 folds at 3 days, P = 0.115). These verified again that rhTGF-β1-enhanced IL-12 secretion promoted HCF migration.
Effect of conditioned medium from IL-12-silenced HCFs on cell migration. (A) A scheme depicting the location of image capture in a culture insert in a six-well culture dish and the area surrounding culture inserts remained cell-free. After removing the culture insert, 2 mL ACM from the HCFs with different treatments as described in Figure 6 were used to incubate HCFs. (B) HCF migration incubated with the conditioned media was monitored, and the images were captured at 3 dpi. HCF migration was performed in different cell sources (n = 3). (C) The number of migratory HCFs that passed the dotted line in (B), which indicates original ibidi insert margin, was counted. The quantitative data represent mean ± SD of the individual experiments performed in different cell sources (n = 3). The difference among the eight groups (panels a–h) were analyzed by 1-way ANOVA followed by Tukey's HSD. Numbers represent P values between the two indicated groups.
Effect of conditioned medium from IL-12-silenced HCFs on cell viability. Cell viability of the ACM-incubated HCFs was determined and calibrated to the individual viability on day 0 at the indicated times. The ACM from the HCFs with different treatments was described in Figure 6. The quantitative data represent mean ± SD of the individual experiments performed in different cell sources (n = 3). Differences in the relative changes in cell viability were analyzed by 1-way ANOVA followed by Tukey's HSD. Numbers represent P values between the two indicated groups.
We demonstrated for the first time that 74.1% of the isolated primary HCFs could secrete proinflammatory cytokine IL-12 in vitro. This secretory IL-12 enhanced both viability and migration of HCFs. TGF-β1 enhanced IL-12 expression in 81.5% primary HCFs (in all the 74.1% secretory group HCFs and in 7.4% nonsecretory group). In addition, the TGF-β1 related expression could be suppressed by MOX. These suggest that corneal stromal cells participate actively in the modulation of corneal inflammation and cellular homeostasis in corneal wound healing via secreting IL-12.
Our results indicate an interindividual variation in the production and TGF-β1 regulation of IL-12 in noninflammatory cell; that is, corneal fibroblast. IL-12 is involved in antigen presentation and corneal inflammation.13–15 As IL-12 is also involved in delayed specific donor allograft rejection29 and that TGF-β signaling inhibition is an emerging strategy for cancer therapy,30,31 whether this TGFβ related IL-12 modulation found in our current study represents a new therapeutic approach in organ/tissue transplantation awaits further investigation. Theoretically, it would be more beneficial in high IL-12 secretory group. It implies that the variability in IL-12 expression found in our experiments contributed at least partially to the prediction of variation in inflammation in corneal infection and/or following surgery. In contrast, for those subjects that do not respond to TGF-β, IL-12 related inflammation and/or rejection could be of less concern postoperatively. This information is beneficial in the era of personalized medicine.
Additionally, as immunosenescence is associated with alteration in inflammation32 and natural killer cells function in the elderly33 and that IL-12 is associated with natural killer cells regulation, it is inspiring to clarify whether there is an age-related difference in IL-12 secretion. We found that there was no significant difference in the age between the IL-12 secretory and nonsecretory group. This indicates that age factor is less important than interindividual genetic variation in the management of corneal infection and/or wound healing regarding IL-12.
Our results demonstrated that 2 ng/mL rhTGF-β1 promoted HCF proliferation in complete culture medium (10% FBS/DMEM). This proliferation would confound cell migration as proliferation could force cell migration. We thus used serum-less media (0.1% FBS/DMEM) in the migration experiment. However, HCFs migrated very slowly in serum-less culture medium compared with the same cells in complete culture medium, we hence used higher IL-12 concentration; that is, 10 ng/mL, in cell migration assay. For parallel comparison, we used rhTGF-β1 10 ng/mL. We found that both IL-12 and TGF-β1 enhanced HCF proliferation (Figs. 4, 8). However, only IL-12, but not TGF-β1, promoted HCF migration (Fig. 5). TGF-β1 promoted cell migration indirectly by increasing IL-12 secretion, which subsequently enhanced HCF migration (Fig. 7C). This is different from previous studies that showed TGF-β1 promoted cell migration in HCFs34,35 and dermal fibroblasts.36 The difference could be evidenced by our experiments using a high concentration of rhTGF-β1 to incubate the isolated HCFs. Our results revealed that 2 ng/mL rhTGF-β1 treatment increased the number of viable HCFs (Fig. 8). If rhTGF-β1 could also promote HCF migration, there should have been an enhanced HCF migration front when rhTGF-β1 was added to the culture media, no matter how many cells were seeded in the culture dish. However, there was no enhanced HCF migration front when HCFs were seeded only in the center of the ibidi culture insert (Fig. 5), although the culture media did contain sufficient rhTGF-β1; that is, 10 ng/mL. This confirmed that rhTGF-β1 alone could not promote HCF migration. In addition, if the HCFs could secrete IL-12 to promote HCF migration indirectly, sufficient number of HCFs would be needed to secrete enough amount of IL-12. This assumption is also confirmed when rhTGF-β1 was added to the 6-cm HCF culture dish in our experiment. The amount of the rhTGF-β1-stimulated IL-12 secretion from the large number of HCFs was sufficient to advance the HCF migration front (Fig. 7B). We thus confirmed that TGF-β1 could indirectly enhance HCF migration via increasing the IL-12 secretion.
In addition to demonstrating that MOX modifies corneal fibroblast-to-myofibroblast differentiation,8 we further illustrated for the first time that MOX reduced IL-12 expression in HCFs concentration-dependently. Our results are clinically relevant and indicate a potentially beneficial anti-inflammatory effect of the topically applied MOX for two reasons. First, the suppression of IL-12 secretion was significant at MOX concentration of 10 μg/mL and higher in the presence of rhTGF-β1. MOX concentration in the cornea is 18.6 ± 9.66 μg/g at 10 minutes after one drop of topical application37 and achieves 2.1 times higher concentrations with repeated topical instillation.38 Since there is a TGF-β1 surge following corneal wounding27 and/or infection, the difference in IL-12 inhibition at MOX concentration of greater than 10 μg/mL in the presence of rhTGF-β1 reflect true clinical scenario. Second, HCFs secrete IL-12 in the presence of lipopolysaccharide stimulation39 and following refractive surgery.15 MOX is normally applied after corneal surgery and/or infection, indicating the current practice of MOX application in a high IL-12 concentration environment. The result of MOX inhibiting IL-12 secretion echoes previous study demonstrating the immunosuppressive effects of ciprofloxacin during human mixed lymphocyte reaction via suppressing the IL-12 production in monocytes.40 Our results thus facilitate understanding of current MOX anti-inflammatory mechanism, in addition to previous antimicrobial and antifibrotic mechanisms.8 However, the beneficial anti-inflammatory effect should be more noticeable in individuals with high baseline IL-12 secretion. MOX might potentially benefit corneal wound healing by prohibiting myofibroblast differentiation8 and by decreasing inflammation via inhibiting TGF-β-induced IL-12 secretion as shown in this study. On the other hand, MOX inhibited HCF migration, which is necessary for wound healing. MOX also diminished HCF proliferation, which is also necessary in stromal repopulation when there is cell depletion. The advantages and disadvantage in wound repair should thus be balanced in clinical application.
Another limitation of our study is that we estimated secretory IL-12 via equation calculation from digitized immunoblot instead of direct ELISA measurement due to the lack of availability of appropriate antibody. However, we generated an equation of the IL-12 standard curve for every experiment in our experiment design to reduce the variation in IL-12 concentration estimation. We suggest that our results of TGF-β1 and MOX related IL-12 alteration revealed a reliable conclusion. Confirming the moxifloxacin-related inhibition of IL-12 secretion using other fluoroquinolones is expected before generalization this wound healing modulation phenomenon.
Supported by Far Eastern Memorial Hospital (FEMH-2012-C-001, FEMH-101-2314-B-418-006-MY3, FEMH-105-2314-B-418-009) and Ministry of Science and Technology (MOST-101-2314-B-418-006-MY3, MOST-105-2314-B-418-009).
Chen TC, Chang SW. Effect of mitomycin C on IL-1R expression, IL-1-related hepatocyte growth factor secretion and corneal epithelial cell migration. Invest Ophthalmol Vis Sci. 2010; 51: 1389–1396.
Kobayashi T, Shiraishi A, Hara Y, et al. Stromal-epithelial interaction study: the effect of corneal epithelial cells on growth factor expression in stromal cells using organotypic culture model. Exp Eye Res. 2015; 135: 109–117.
Wilson SL, Yang Y, El Haj AJ. Corneal stromal cell plasticity: in vitro regulation of cell phenotype through cell-cell interactions in a three-dimensional model. Tissue Eng Part A. 2014; 20: 225–238.
Arranz-Valsero I, Soriano-Romani L, Garcia-Posadas L, Lopez-Garcia A, Diebold Y. IL-6 as a corneal wound healing mediator in an in vitro scratch assay. Exp Eye Res. 2014; 125: 183–192.
Torricelli AA, Santhanam A, Wu J, Singh V, Wilson SE. The corneal fibrosis response to epithelial-stromal injury. Exp Eye Res. 2016; 142: 110–118.
Ljubimov AV, Saghizadeh M. Progress in corneal wound healing. Prog Retin Eye Res. 2015; 49: 17–45.
Singh V, Jaini R, Torricelli AA, et al. TGFbeta and PDGF-B signaling blockade inhibits myofibroblast development from both bone marrow-derived and keratocyte-derived precursor cells in vivo. Exp Eye Res. 2014; 121: 35–40.
Chen TC, Chang SW, Wang TY. Moxifloxacin modifies corneal fibroblast-to-myofibroblast differentiation. Br J Pharmacol. 2013; 168: 1341–1354.
Lee HW, Chung SH, Moon CM, et al. The correlation of serum IL-12B expression with disease activity in patients with inflammatory bowel disease. Medicine (Baltimore). 2016; 95: e3772.
Hunter CA. New IL-12-family members: IL-23 and IL-27, cytokines with divergent functions. Nat Rev Immunol. 2005; 5: 521–531.
Kawabe K, Lindsay D, Braitch M, Fahey AJ, Showe L, Constantinescu CS. IL-12 inhibits glucocorticoid-induced T cell apoptosis by inducing GMEB1 and activating PI3K/Akt pathway. Immunobiology. 2012; 217: 118–123.
Jun AS, Cope L, Speck C, et al. Subnormal cytokine profile in the tear fluid of keratoconus patients. PLoS One. 2011; 6: e16437.
Hu J, Hu Y, Chen S, et al. Role of activated macrophages in experimental Fusarium solani keratitis. Exp Eye Res. 2014; 129: 57–65.
Frank GM, Divito SJ, Maker DM, Xu M, Hendricks RL. A novel p40-independent function of IL-12p35 is required for progression and maintenance of herpes stromal keratitis. Invest Ophthalmol Vis Sci. 2010; 51: 3591–3598.
Leonardi A, Tavolato M, Curnow SJ, Fregona IA, Violato D, Alio JL. Cytokine and chemokine levels in tears and in corneal fibroblast cultures before and after excimer laser treatment. J Cataract Refract Surg. 2009; 35: 240–247.
Tomcik M, Zerr P, Palumbo-Zerr K, et al. Interleukin-35 is upregulated in systemic sclerosis and its serum levels are associated with early disease. Rheumatology (Oxford). 2015; 54: 2273–2282.
Bauler TJ, Chase JC, Bosio CM. IFN-beta mediates suppression of IL-12p40 in human dendritic cells following infection with virulent Francisella tularensis. J Immunol. 2011; 187: 1845–1855.
Chung JL, Seo KY, Yong DE, et al. Antibiotic susceptibility of conjunctival bacterial isolates from refractive surgery patients. Ophthalmology. 2009; 116: 1067–1074.
Ortega-Usobiaga J, Llovet-Osuna F, Djodeyre MR, Llovet-Rausell A, Beltran J, Baviera J. Incidence of corneal infections after laser in situ keratomileusis and surface ablation when moxifloxacin and tobramycin are used as postoperative treatment. J Cataract Refract Surg. 2015; 41: 1210–1216.
Haripriya A, Chang DF, Namburar S, Smita A, Ravindran RD. Efficacy of intracameral moxifloxacin endophthalmitis prophylaxis at Aravind Eye Hospital. Ophthalmology. 2016; 123: 302–308.
Rudnisky CJ, Wan D, Weis E. Antibiotic choice for the prophylaxis of post-cataract extraction endophthalmitis. Ophthalmology. 2014; 121: 835–841.
O'Brien TP, Arshinoff SA, Mah FS. Perspectives on antibiotics for postoperative endophthalmitis prophylaxis: potential role of moxifloxacin. J Cataract Refract Surg. 2007; 33: 1790–1800.
Pettey JH, Mifflin MD, Olson RJ. Study of the acute effects of povidone-iodine on conjunctival bacterial flora. J Ocul Pharmacol Ther. 2015; 31: 627–630.
Beisswenger C, Honecker A, Kamyschnikow A, Bischoff M, Tschernig T, Bals R. Moxifloxacin modulates inflammation during murine pneumonia. Respir Res. 2014; 15: 82.
Eye Bank Association of America. 2011 Eye Banking Statistical Report. Washington, DC: Eye Bank Association of America; 2012.
Santacruz C, Linares M, Garfias Y, et al. Expression of IL-8, IL-6 and IL-1beta in tears as a main characteristic of the immune response in human microbial keratitis. Int J Mol Sci. 2015; 16: 4850–4864.
Uwaydat S, Jha P, Tytarenko R, et al. The use of topical honey in the treatment of corneal abrasions and endotoxin-induced keratitis in an animal model. Curr Eye Res. 2011; 36: 787–796.
Park GB, Kim D, Kim YS, et al. The Epstein-Barr virus causes epithelial-mesenchymal transition in human corneal epithelial cells via Syk/src and Akt/Erk signaling pathways. Invest Ophthalmol Vis Sci. 2014; 55: 1770–1779.
Verma ND, Hall BM, Plain KM, et al. Interleukin-12 (IL-12p70) promotes induction of highly potent Th1-like CD4(+)CD25(+) T regulatory cells that inhibit allograft rejection in unmodified recipients. Front Immunol. 2014; 5: 190.
Neuzillet C, Tijeras-Raballand A, Cohen R, et al. Targeting the TGFbeta pathway for cancer therapy. Pharmacol Therapeutics. 2015; 147: 22–31.
Colak S, Ten Dijke P. Targeting TGF-beta signaling in cancer. Trends Cancer. 2017; 3: 56–71.
Brown BN, Haschak MJ, Lopresti ST, Stahl EC. Effects of age-related shifts in cellular function and local microenvironment upon the innate immune response to implants. Semin Immunol. 2017; 29: 24–32.
Pera A, Campos C, Lopez N, et al. Immunosenescence: implications for response to infection and vaccination in older people. Maturitas. 2015; 82: 50–55.
Chang Y, Wu XY. The role of c-Jun N-terminal kinases 1/2 in transforming growth factor beta(1)-induced expression of connective tissue growth factor and scar formation in the cornea. J Int Med Res. 2009; 37: 727–736.
He J, Bazan HE. Epidermal growth factor synergism with TGF-beta1 via PI-3 kinase activity in corneal keratocyte differentiation. Invest Ophthalmol Vis Sci. 2008; 49: 2936–2945.
Sapudom J, Rubner S, Martin S, Thoenes S, Anderegg U, Pompe T. The interplay of fibronectin functionalization and TGF-beta1 presence on fibroblast proliferation, differentiation and migration in 3D matrices. Biomater Sci. 2015; 3: 1291–1301.
Domingos LC, Moreira MV, Keller KM, Viana FA, Melo MM, Soto-Blanco B. Simultaneous quantification of gatifloxacin, moxifloxacin, and besifloxacin concentrations in cornea and aqueous humor by LC-QTOF/MS after topical ocular dosing. J Pharmacol Toxicol Methods. 2017; 83: 87–93.
Chung JL, Lim EH, Song SW, et al. Comparative intraocular penetration of 4 fluoroquinolones after topical instillation. Cornea. 2013; 32: 1046–1051.
Kimura K, Orita T, Nomi N, Fujitsu Y, Nishida T, Sonoda KH. Identification of common secreted factors in human corneal fibroblasts exposed to LPS, poly(I:C), or zymosan. Exp Eye Res. 2012; 96: 157–162.
Katsuno G, Takahashi HK, Iwagaki H, et al. The immunosuppressive effects of ciprofloxacin during human mixed lymphocyte reaction. Clin Immunol. 2006; 119: 110–119.


text:
package org.apache.jackrabbit.oak.segment.util;

import static org.junit.Assert.assertArrayEquals;
import static org.junit.Assert.assertEquals;
import static org.junit.Assert.fail;

import java.io.IOException;
import java.nio.charset.StandardCharsets;
import java.util.UUID;

import org.junit.Test;

public class CharsetEncodingUtilsTest {

    private static char[] SURROGATE_PAIR = Character.toChars(0x1f4a9);

    @Test
    public void encodeValid() throws IOException {

        // a a-umlaut euro plane-1-char
        String test = "a \u00E4 \u20ac " + new String(SURROGATE_PAIR);

        byte[] withStringClass = test.getBytes(StandardCharsets.UTF_8);
        byte[] withUtilsClass = CharsetEncodingUtils.encodeAsUTF8(test);
        assertArrayEquals(withStringClass, withUtilsClass);
        assertEquals(test, new String(withUtilsClass, StandardCharsets.UTF_8));
    }

    @Test
    public void encodeInValid() {

        // a a-umlaut euro plane-1-char, second char in surrogate pair missing
        String test = "a \u00E4 \u20ac " + SURROGATE_PAIR[0];

        try {
            CharsetEncodingUtils.encodeAsUTF8(test);
            fail("expected encoding to fail");
        } catch (IOException expected) {
            // expected
        }
    }

    @Test
    public void encodeMultiThreaded() throws InterruptedException {

        int tc = 20;

        Thread[] threads = new Thread[tc];

        for (int i = 0; i < tc; i++) {
            threads[i] = new Thread(new Runnable() {
                @Override
                public void run() {
                    // encode and decode 100 random UUID strings
                    for (int j = 0; j < 100; j++) {
                        String test = UUID.randomUUID().toString();
                        String roundtripped = null;
                        try {
                            byte[] bytes = CharsetEncodingUtils.encodeAsUTF8(test);
                            roundtripped = new String(bytes, StandardCharsets.UTF_8);
                        } catch (IOException exignored) {
                        }
                        assertEquals(test, roundtripped);
                    }

                }
            });
        }
        for (int i = 0; i < tc; i++) {
            threads[i].start();
        }
        for (int i = 0; i < tc; i++) {
            threads[i].join();
        }
    }
}

redpajama_set_name: RedPajamaGithub


On July 10th, I was invited to K-Palette's Eyes On You Beauty Bloggers' Workshop by The Sample Store.
The event was held at Fika - Swedish Café and Bistro. Whimsical and cozy, you and your lovelies can find it on the corner of Beach Road and Arab Street (No. 257 Beach Road/ Arab Street).
I have been waiting for this event since I got the invitation in the middle of June!
I went with Fel and bumped into Lynn, who just got back from the States!
Since I made the switch from pencil eyeliners to liquid ones, the 24H Real Lasting Eyeliner is all that I have been using!
Sharp and thin, I can decide on the thickness of my liner with ease and it doesn't go all flaky on me. It doesn't smudge on my oily eyelids too, which is the main reason why I ditched pencil and went liquid.
The Lasting 2Way Eyebrow is K-Palette's newest product to hit our shores!
There's Mr. Soh in the top picture. He's the head of Japalang. Sharon's the one with the mike and Ariel's the one in the white-dotted dress. Mariko-san, in the striped tee, was the makeup artist for the day!
Mariko-san taught us how to do Glamorous and Kawaii looks. Check out my friends Eunice and Mich sporting the 2 looks here!
The latest makeup fad in Japan is to draw bottom lashes right at the corner to give your lashes a fuller look (Now you know!)!


The PrivateBank's mission is to advance the goals of its clients through compelling financial solutions and to strengthen our communities by supporting organizations that make a difference in our cities, towns and neighborhoods. Our relationship-based business model means our is dedicated to knowing our clients and what matters most to them. The PrivateBank provides middle market commercial and commercial real estate companies with customized relationship banking services including financing, treasury management, capital markets and liquidity solutions. The private wealth team works with the owners and executives of commercial clients as well as other high-net-worth families to help with asset growth, preservation and transition. Additionally, its branch network provides personal, small business and mortgage banking services to the communities it serves.


Priority boarding with access to 1st come 1st Serve open seating, reserved private area, dedicated server, plus upgraded Glow Party Favors.
Admits 5 with all the VIP Perks at a special Group Rate.
From the creators of Pier Pressure... Halloween Night! Saturday, October 26th, 2019 -- Prepare for the ultimate Halloween party cruise that will be transformed into a haunting Titanic, filled with over 300+ ghosts & ghouls raised from the dead of the historic Titanic that sank over a hundred years ago.
Don't be afraid to come aboard the most magnificent and spacious yacht 'The Magic Spirit'. We'll rock this boat with the best club mixes on the planet.
- This is the biggest Yacht Party in Canada this Halloween!
- Over 300 Titanic party people in attendance having the time of their lives!
Costumes are not required but strongly encouraged. All mask will need to be removed prior to entering for verification of your ID. No weapons will be tolerated including toy: swords, knifes and/or guns regardless if they do not appear to be real or not. We reserve the right to refuse service to anyone or confiscate any props to a costume that may deem inappropriate or dangerous.
This isn't like any other cruise or event. It's Vancouver Nightlife's mission is to make sure our 'Pier Pressure' Yacht Party is a perfect one for you and all your friends. Join us on the ultimate Party Cruise! Call us today if you have any questions or need assistance: 604.283.6601.
:| All Sales are final. NO Refund or exchanges. Prices are subject to change while supplies last.


Size: 1.320mq - 14.200 sq.ft.
Pisa is one of the most important cities in Tuscany and well-known places in the world because of its famous symbol, the leaning tower, known as Pisa Tower. Situated in the center of the Piazza deiMiracoli, Miracle Square, it is a really magic monument that attracts millions of tourists yearly. It is also a city of Alma Mater with its three important Italian universities: the University if Pisa, the Sant'Anna School of Advanced Studies and the ScuolaNormaleSuperiore. The city is proud by its rich history, it is used to be one of the four marittime republics. Nowadays it can also be proud of its modern international Galileo Galiley Airport that connects it to anywhere in the world.
Pisa is surrounded by charming Tuscan countryside. Having a promenade through the narrow streets of the city center or visiting interesting places close to the town like Certosa di Calci is a pleasure that never ends. The beautiful spa place CascianaTerme proposes relaxing and healthy baths for the well-being of the entire family. Different activities such as hiking, biking and horse riding make the life in Pisa full and exiting.


Amazon Internal Press Release in Product Management.
Last year I start using Amazon Internal Press Release and it improved my approach to product development and product management.
Is your product development process creating more problems than it's solving?
Product Development Process: I've been in all types of startups from where there's no product development process at all, to a startup that established a programming military machine as their go-to product process, and a startup where I implemented a product development process myself.
When product becomes a military procedure.
Something interesting is that each startup, each team, and each product is a new world. Even though each of them operates differently, there's always something that joins them: The Process. Because even if you have no established process, at the end it's ad hoc process or an on-demand one.
Building a product is a cross-functional endeavor, as PMs we must deal with everything and everyone, we are the ajonjolí de todos los moles.
The Product's outcome is on the product manager, in other words, the product manager is ultimately responsible for the success or failure of a product, and I say no one and everyone, because, we PMs don't have any authority over any of the people involved in the collective effort.
So we are no one's boss, but at the end, if the product fails, it's our responsibility; sounds like a Catch 22 to me. That's why the Product Manager should always have a card up their sleeve: Process.
Let me explain with an example; many startups and product teams have adopted Objective and Key Results ("OKRs"): please read my article about OKRs if you don't know what they are, and also so you can establish better NYE resolutions.
OKRs, are a great example of a process that establishes Communication, Autonomy, and Execution.
Let's say a company's main Goal aka Objective is to increase brand awareness, that would require a couple of key results; maybe improving the landing page, adding a product tour, increasing the signup funnel by xyz%, and so on.
What's interesting about this example, is that you are i) communicating at the company or team level the main objective; but you leave ii) autonomy to the team of how to solve this, and how such iii) execution will be measured.
So any process, should follow such schema, unless you are reading this in Fort Bragg and you want to apply it with your squad, be my guest but demanding is not the same as communicating.
So moving back to Product Development Process, how do we establish certainty, replace chaos with order, in a such a fast changing world? Maybe a process that solved yesterday's problems could block a team from developing tomorrow's solutions.
1.- It takes twice the planning & execution of the process, than the actual execution of the Product.
Sometimes it's easy to underestimate the time and cost of executing a process with the product & Dev team. If the managers are asking for 1 Pagers, user flows, lowframes, high frames, wireframes, Mocks, PRD (product requirement doc), Postmortems, Sprint Planning, Mid-Sprint Review, costing, burn-down charts post this and that. You could get lost more on the planning and reviewing than actually executing the product.
All product managers have the same 24 hours, and it's up to the best PM's using their time talking to the customer rather than going through a bureaucratic development process.
Product is King, but the User is the King of Kings, and if the product development process is not centered towards her you are just wasting yours team's time.
The life-cycle of product is dependent on its iterations, new incoming variables, and in & out factors, and that's how the Product process should be.
We must always allow the process to murmurate in a constant flow ?
If you enjoyed this post, please hit the "recommend" ❤ button below. Thanks, and remember to follow me on Instagram.
I remember when I was in business class and my professor said: "Cash is King". This meant good cash flow and very good financial health inside a company.
The Product is King because a great product is created by an amazing team, based on outstanding customer insight, clear data understanding, and yes sometimes luck along the way.
Product is the realm where things actually happen because it is built to focus on just one thing, to bring a solution to an existing problem.
The hardest thing is getting your product to work on a global scale; marketing, sales, and money will come in their own time if the product excels. We should create a dependable commodity product, something that people love so much. Something that people cannot live without.


So are you sick of YouTube like me. Well maybe Revver isn't the answer as it's all about making money from your videos. But then if your creating video content and you want some monetary compensation for doing it then it probably is the answer for you. Otherwise Id probably tell you to continue wastering Youtube's bandwidth.
As you can see from the video YouTube is playing catchup.
This entry was posted in Internet / Online on May 2, 2008 by JAB_au.


text:
CHRISTIAN HALL
Born 1973, Mona Vale, New South Wales
Lives Adelaide, South Australia
Working across furniture and industrial design, and sculpture and jewellery, each piece Christian Hall makes is designed to inspire a moment of delight. His meticulous designs focus not only on form, but on physicality, resulting in sculptural objects which seamlessly combine hand-crafting skills with a sleek industrial-looking finish.
Graduating from Sydney College of the Arts in 1998 with a Bachelor of Visual Arts, First Class Honours, Hall began his career in jewellery and object design. Going onto complete his Masters in Visual Arts in 2003, Hall has been the creative director at JamFactory Jewellery & Metal Design Studio since 2009. An accomplished teacher and maker, Hall has exhibited widely across Australia and internationally, in addition to being the recipient of numerous awards and grants.
Mesh Room Divider was born from a commission by a local shared space in Kent Town, Adelaide which hosts a cafe, gallery and design studio, to find a solution to separate the cafe and the design studio, without losing the sense of open space. In the original design Hall used repurposed rebar) to create a floating room divider. In this new version Hall opted for a powder coated mild steel, instead of rebar, while also changing the grid subtly, to diamond shaped pattern, forming a slightly softer aesthetic.
The jewellery making process combines laser cutting and micro-welding with more traditional hand-making techniques and elements of play-like construction. Each unit is made from the same two, flat, interlocking components that are snapped together by hand to sit at right angles to each other. These singular, wire frame structures create a description of three-dimensional space that when multiplied produce works that explore an ambiguity between line and volume.

redpajama_set_name: RedPajamaCommonCrawl


text:
namespace PactNet.Matchers
{
    /// <summary>
    /// Match an integer specifically
    /// </summary>
    public class IntegerMatcher : IMatcher
    {
        /// <summary>
        /// Type of the matcher
        /// </summary>
        public string Type => "integer";

        /// <summary>
        /// Matcher value
        /// </summary>
        public dynamic Value { get; }

        /// <summary>
        /// Initialises a new instance of the <see cref="IntegerMatcher"/> class.
        /// </summary>
        /// <param name="value">Example value</param>
        internal IntegerMatcher(int value)
        {
            this.Value = value;
        }
    }
}


Collect contact details for Building Contractors in California USA.
Collect contact details for (1) Building Contractors, (2) General Contractors and (3) Real Estate Developers in California USA. Need minimum 5,000 businesses with email addresses.
I am interested to do this project. ..............Can start now............................................Lets start. Please contact for further communication.


Join a committee - Help us to help you get better gigs.
Don't be a square. Get hep to the crazy new jazz & latin scene in town!
Got a band but can't get action?
Blowin' in your pad, but can't find the right cats to make the gig?
Earthbound, but cranked on jazz & want to know who's cookin?
We serve our membership by providing and promoting a range of resources for networking among jazz musicians, patrons and aficionados, and venues at which jazz may be heard.


Renovation is a field with low barriers to entry. Anyone can pick up a hammer and claim to be a remodeler. Choose a contractor who has been around for a number of years and who has a substantial number of positive references. Ideally, the contractor should be affiliated with organizations like the National Association of the Remodeling Industry (NARI), the National Association of Home Builders (NAHB), the Home Builders Association of Grand Rapids (HBA). Awards from local or national organizations also speak well of a contractor's experience and credibility.
Past successes are always an important factor in selecting a contractor. Determine if the contractor has completed any projects that are similar in scope to your planned renovation. Check out their portfolios and see if they can personally take you to view any similar completed projects first hand.
Timely, accurate answers from your contractor are the foundation of a successful project and a solid contractor-client relationship. If the contractor doesn't communicate clearly and effectively from the outset, you are more likely to encounter frustrations - both emotional and financial - as the project continues.
Without a detailed, fixed-sum proposal there are no guarantees against budget overruns that could cost you greatly. Knowing exactly what the contractor is bidding on will help you determine if your concept matches up with the proposed work and limits the need for change orders for work that the contractor may claim was not included in their bid. We do not recommend "cost plus" contracts, as they leave little incentive for the contractor to remain within your budget.
Be certain that any contractor you choose to work with carries a General Liability insurance policy of at least $1 million. Confirm that they can offer Builders Risk insurance if necessary. Request a paper copy of the contractor's insurance certificate for your records.
When making your final selection for a renovation contractor, be sure ask for a minimum of 5 references and do the legwork of checking them. A contractor without references is one to be avoided.
The work performed by the contractor is something you will have to live with for many years. Be certain that there are protections and remedies in place should something go wrong down the road. The contract should contain a warranty description that states clearly what is covered.
Without a strong commitment to a completion date, you could find yourself unable to return to your home when planned, which could cause you additional financial hardship as well as headaches in determining where to reside until the work is completed. A reputable contractor will provide you with a firm completion date.


text:
William Farrington Aldrich (* 11. März 1853 in Palmyra, Wayne County, New York; † 30. Oktober 1925 in Birmingham, Jefferson County, Alabama) war ein US-amerikanischer Politiker (Republikanische Partei). Er war der Bruder von Truman H. Aldrich und Ururgroßvater von William J. Edwards, ebenfalls Kongressabgeordnete für Alabama.

Werdegang
William Farrington Aldrich besuchte die öffentliche Schule in seiner Heimatstadt. Dann zog er 1865 mit seinem Vater nach New York City, wo er mehrere Schulen besuchte und schließlich 1873 an Warrens Military Academy in Poughkeepsie (New York) graduierte. Im nachfolgenden Jahr zog er nach Alabama, wo er Bergbau- und Fabrikgeschäften nachging. Ferner erbaute er die Stadt Aldrich, die seinen Namen trägt.

Aldrich verfolgte ebenfalls eine politische Laufbahn. Er hat zweimal die Wiederwahl von Gaston A. Robbins (54. und 56. US-Kongress) und einmal die Wahl von Thomas S. Plowman (55. US-Kongress) erfolgreich in den US-Kongress angefochten. Aldrich diente im US-Repräsentantenhaus vom 13. März 1896 bis zum 3. März 1897, dann vom 9. Februar 1898 bis zum 3. März 1899 und schließlich vom 8. März 1900 bis zum 3. März 1901. Er lehnte 1900 eine erneute Kandidatur für den US-Kongress ab.

Danach war er als Redakteur, Eigentümer und Verleger der Birmingham Times tätig. Er nahm 1904 an der Republican National Convention in Chicago teil. Ferner beschäftigte er sich bis zu seinem Tod 1925 mit der Erschließung der Rohstoffvorkommen. Sein Leichnam wurde eingeäschert und in der Familiengruft auf dem Rock Creek Cemetery in Washington, D.C. beigesetzt.

Weblinks
William F. Aldrich auf der Website von www.archive.org

Mitglied des Repräsentantenhauses der Vereinigten Staaten für Alabama
Mitglied der Republikanischen Partei
US-Amerikaner
Geboren 1853
Gestorben 1925
Mann

redpajama_set_name: RedPajamaWikipedia


Traded on coins and $0,01. The points are normal, especially for the price. But the store the case-considers time for free and it goes, he would learn from other sellers (VEITHDIA, 20/20), which do not suffer.
It Is really cool sunglasses! The quality is excellent! Complete accessories for storage and maintenance. Clever changed the desired color!
Great product and the store! Send fast, came too quickly. Track to track. All happy.
It 'S excellent points! Complete with case and real polarized filters. All advice!


Raazi is a 2018 Indian spy suspenseful story film in which the Alia bhatt and Vicky Kaushal has played the leading role .Director of movie is Meghna Gulzar and producer of movie is Vineet Jain, Karan Johar, Hiroo Yash Johar and Apoorva Mehta under the banner of Dharma Productions. Raazi is title for a second weekend of 20 crore net plus and it might turn out to be the second utmost second weekend of the year after Padmaavat.
Business of Raazi till date is as follows.


text:
Richard Rankin
January 4 Actor#19
Capricorn Named Richard#9
Last Name Rankin#4
Most well known for his work on the Scottish television sketch comedy program Burnistoun and for his portrayal of Captain Thomas Gillan on the BBC One historical series The Crimson Field, Rankin is also notable for his recurring roles on The Syndicate and American Odyssey.
After attending Langside College in Glasgow, Scotland, he made his television acting debut in a 2006 episode of VideoGaiden.
His film credits include roles in The House of Him (2014) and Adam Jones (2015).
Born Richard Harris, he is more widely known by his stage name of Richard Rankin (Rankin being his mother's maiden name). He and his brother and fellow television actor, Colin Harris, were raised in Rutherglen, Scotland.
In 2010, he appeared with Alex Norton in an episode of the crime series Taggart.
Richard Rankin Popularity
Richard Rankin Is A Member Of
The Syndicate
From Darkness
Richard Rankin Fans Also Viewed


We are excited to announce our NEW website! We hope you all love it. Take a peek at the new site and new departments on the left hand side to help you easily navigate our colorful selection of products!
Sign-up for our E-News HERE.
Request a copy of our SUMMER 2011 catalog (yet to be mailed) HERE.
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It looks beautiful. I love the colors and the design.


93 points David Schildknecht, Wine Advocate, "Diversely-sourced - its Merlot blended with 35% Cabernet Sauvignon- the Betz 2010 Clos de Betz emphasizes black pepper, pungently resinous herbs, and arbor vitae, characteristics I imagine are in part a reflection of its 7% of Petit Verdot, that grape being demonstrably (I'm tempted to say notoriously ) efficacious. Cassis, huckleberry, and walnut offer bitter-edged but sappy fruit satisfaction; a crushed stone undertone adds a sense of intrigue; and mineral salts savory saliva-inducement in a gripping, palate-staining finish. This ought to be worth following for at least the better part of a decade. "


text:
Why use rapid capture time-lapse?
11th September 2017 - Posted by: Wallis Seaton
Time-lapse is often associated with rendering long-term work in a neat and manageable visual sequence. For projects taking place over a much shorter timeframe, however, rapid interval time-lapse offers a means of capturing faster-paced works over a matter of days, weeks, and months.
Rapid capture means that the camera system takes images at a faster pace than is usually expected for long-term works.
One image every 15 minutes over the course of an average working day from 8am to 5pm, would be an ideal set-up for construction works of one year or more. For faster-paced projects, though, such as a one-day event or an office fit-out over a few weeks for example, the camera will need to be equipped to capture much more frequently.
Indeed, the term 'rapid capture' takes its name from the greater number of images needed to produce a time-lapse video of a worthwhile length.
Capturing every detail
Internal works benefit greatly from rapid interval photography as any visual changes that occur within a particular enclosed space are significantly emphasised.
Expansion of Sky Bet's head office in Leeds provides an excellent example of what can be achieved with this technique. Tracking 15-weeks worth of intense activity, the quicker frequency of capture means that every detail of progress was picked up for a concise but comprehensive minute-long narrative.
Spaces set up temporarily for special events and leisure activities also benefit greatly from rapid capture as construction and assembly of such spaces are often limited to hours and days.
This was the case when capturing the prestigious Horse Guards Parade St James' Park and The Mall in London, where a temporary building was constructed on the Parade Ground. Rapid capture enabled extensive coverage of the lavish decoration of this interior.
The construction of the British Film Foundation's temporary Embankment Garden Cinema also required both internal and external rapid capture.
A 60-second post-produced edit was perfect for sharing on social media platforms, where the visual sequence helped to spread the word of these short-term works as part of special 60th anniversary celebrations for the London Film Festival.
"If you build it they will come…" #LFF pic.twitter.com/eSgTROhGDw
— BFI (@BFI) October 7, 2016
Remote time-lapse monitoring allows for a greater degree of control over rate of capture in external environments, as camera settings can be adjusted as and when is appropriate. This is essential when at the mercy of temperamental weather conditions, like those in Scotland where we captured the erection of the first ever two-tier grandstand at St Andrews.
Stunning combinations
Of course, rapid capture can also be utilised in combination with video and time-lapse in the same sequence.
Different aspects of a project require different things from the camera and so it is not always appropriate to use the same rate of capture throughout.
Above: rapid interval time-lapse was effectively used to capture everyday activities at Southampton Freight Services.
A fit-out for retailer Vision Express at their flagship Oxford Street store, for example, involved video, time-lapse, and rapid capture in order to draw attention to important details as the shop transformed from a basic skeletal structure into a stylish commercial venue as part of a stunning visual narrative.
Honing in on specific details of internal works may also require more than a fixed camera method. For this behind-the-scenes look at the everyday workings of Southampton Freight Services, for instance, rapid capture was engaged on the ground as well in addition to a fixed position solution.
Even from this handful of examples, then, it is clear that time-lapse is an extremely versatile mode of photography.
And while time-lapse is typically employed to showcase long-term projects where progress unfolds at a steady pace, short-term works can be rendered just as effectively using rapid capture solutions.
Contact us to find out more about what we can offer.
Categories: Construction Event External Internal Off-grid Photography Time-lapseTags: BFIcamera systemsevent monitoringHorse Guards ParadeLondonlong-termpost-productionrapid captureremote captureshort-termSouthampton Freight ServicesSt Andrewstime-lapse videoVision Express
Time-lapsing the Northern Powerhouse: spotlight on Manchester
The importance of a multimedia approach to business


Transformers Series 2 Decepticon Jets. In 1985 Hasbro launched 3 more jets into their line up. These jets were all retools from the series 1 jets. This time the wings were all changed instead of just a new color. The weapons were changed on Ramjet and Dirge while Thrust keeps the series 1 weapons, just colored differently.
The Series 2 Jets are made up of Thrust, Ramjet, and Dirge. All three are nicknamed the coneheads for the way they appear in the cartoon. Although not show as such on the box art or the instructions most of us will transform them with the nose cone of the jet facing up to make that conehead look.
Shown here are very nice examples from my personal collection. The Thrust is graded a high 85 by AFA while Ramjet and Dirge are both graded a 90 overall by AFA.


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text:
Benedict Cumberbatch Will Play Satan In Amazon's Good Omens
By Laura Hurley published 14 February 19
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Benedict Cumberbatch is returning to the small screen, but not to tackle his iconic role as Sherlock Holmes. (Not yet, at least.) No, the actor has landed a role in the upcoming adaptation of Neil Gaiman's Good Omens, which premieres on Amazon later this year. Cumberbatch is on board to play Satan.
Yes, the Sherlock and Doctor Strange star has signed on to play Satan for Good Omens, but don't expect to see him in the flesh. Benedict Cumberbatch will actually be voicing Satan. While that might disappoint Cumberbatch fans who would rather see him in live-action, there's a very good reason why he won't appear as Satan.
Neil Gaiman, who wrote the Good Omens novel, revealed at the TCA press tour panel for the new show (via Deadline) how Benedict Cumberbatch will be incorporated as Satan:
It's a giant animated Satan. He turns up and we needed a performance that works. So we found a young British help that needed a lot of direction from Douglas… Benedict Cumberbatch.
Benedict Cumberbatch is a versatile actor who can do many things on screen, but appearing as a giant animated Satan isn't really within any actor's wheelhouse. While Neil Gaiman didn't give away exactly what viewers should expect Cumberbatch's Satan to look like, he should be equal to the role. Cumberbatch has plenty of experience with voice-acting. I have to wonder how he felt when he learned he was deemed the right man for the job of Satan!
Prospective viewers shouldn't count on seeing lots of Benedict Cumberbatch's Satan on Good Omens. Satan will only appear in the sixth episode, which is also the season finale. Still, I'm guessing what screentime he has will be compelling. Neil Gaiman and Co. needed a memorable and formidable actor to play Satan because they landed a memorable and formidable actress to play God.
Frances McDormand, who won an Academy Award and a Golden Globe in 2018 for her performance in Three Billboards Outside Ebbing, Missouri, is the voice of God on Good Omens. Will Benedict Cumberbatch be a worthy equal as Satan?
Benedict Cumberbatch and Frances McDormand are only two of the star-studded cast of Good Omens. David Tennant (Doctor Who) and Michael Sheen (Masters of Sex) star as a loose-living Demon and a fussy Angel, respectively. They team up to prevent the Armageddon, as they had become fond of life on Earth. Also appearing on Good Omens are Jon Hamm, Nick Offerman, Jack Whitehall, Miranda Richardson, Adria Arjona, Michael McKean, Anna Maxwell Martin, Mireille Enos, and another Doctor Who alum.
The very good news is that Amazon has finally given Good Omens a premiere date. The show will hit the web for Amazon subscribers on Friday, May 31. All things considered, 2019 is already shaping up to be a big year for Amazon. Jack Ryan was just renewed for a third season, a superhero series is on the way, and more. For some upcoming Amazon premiere dates, swing by our 2019 midseason schedule.
Leonardo DiCaprio Weighs In On Netflix's Wild Don't Look Up Ending, And Why It Needed To Play Out The Way It Did


text:
<!DOCTYPE html>

    <meta charset="utf-8">
    <title>BiwaScheme</title>
    <link href="/website/css/screen.css" rel="stylesheet" type="text/css">
    <link href="https://cdn.jsdelivr.net/npm/jquery.terminal/css/jquery.terminal.min.css" rel="stylesheet"/>
    <link href="/website/css/prism-coy.css" rel="stylesheet" type="text/css"></script>
    <meta name="viewport" content="width=device-width, initial-scale=1.0"/>

Features

Getting Started

Reference

FAQ

Other docs

List of functions

Basic types

Numeric types

Control structure

Macro

Advanced types

Utilities

Syntax

JavaScript language interface

Browser functions

Node.js

BiwaScheme JavaScript API

Basic types

List

length append reverse list-tail list-ref map for-each

find for-all exists filter partition fold-left fold-right

remp remove remv remq

memp member memv memq

assp assoc assv assq

cons*

list-sort

(iota count start? step?) (srfi-1)

(list-copy list) (srfi-1)

String

string? make-string string string-length string-ref

string=? string<? string>? string<=? string>=?

substring string-append string->list list->string

string-for-each string-copy

(open-input-string s) (srfi-6)

(open-output-string) (srfi-6)

(get-output-string strport) (srfi-6)

Vector

vector? vector vector-length vector-ref vector-set!

vector->list list->vector vector-fill! vector-map vector-for-each

(vector-append v1 v2...) (vector-copy v) (srfi-43)

vector-sort vector-sort!

Boolean

boolean? boolean= not and or

Pair

pair? null? list?

cons list car cdr set-car! set-cdr!

caar cadr ... cdddar cddddr

Symbol

symbol? symbol->string symbol=? string->symbol

Char

char? char->integer integer->char char=? char<? char>? char<=? char>=?

Numeric types

Integer (1, -2, etc.)

integer? odd? even?

(random-integer n) => 0..n-1 (srfi-27)

Rational (literal not supported yet)

rational?

Real (1.2, -2.3, etc.)

(random-real) => 0.0...1.0 (srfi-27)

Complex // literal not supported yet

complex? make-rectangular make-polar real-part imag-part magnitude angle

Number

number? = < > <= >= zero? positive? negative?

finite? infinite? nan?

max min + - * / abs div0-and-mod0 div mod div0-and-mod0 div0 mod0

numerator denominator floor ceiling truncate round (rationalize)

exp log sin cos tan asin acos atan sqrt exact-integer-sqrt expt

number->string string->number

Control structure

Conditional

if when unless cond case

Loop

do let(named let) dotimes

Exception

raise

Assignment

let let1 let\* letrec letrec\*

Values

values call-with-values let-values let\*-values

(receive <formals> <expression> <body>...) (srfi-8)

apply

eval

eq? eqv? equal?

Macro

BiwaScheme does not have syntax-rules or syntax-case, but has define-macro.

(define-macro (test expr)
  `(if ,expr
    #t
    (print (format "test failed: ~a" (quote ,expr)))))

(test (= 1 2)) ;=> test failed: (= 1 2)

Other macro-related functions:

macroexpand macroexpand-1 %macroexpand %macroexpand-1 gensym

Advanced types

Port (I/O)

Record

define-record-type

Hashtable

make-eq-hashtable make-eqv-hashtable make-hashtable

equal-hash string-hash string-ci-hash symbol-hash

hashtable? hashtable-size hashtable-ref hashtable-set!

hashtable-delete! hashtable-contains? hashtable-update!

hashtable-copy hashtable-clear! hashtable-keys hashtable-entries

hashtable-equivalence-function hashtable-hash-function hashtable-mutable?

Enumeration

define-enumeration

enum-set-member? enum-set-subset? enum-set=? enum-set->list

enum-set-union enum-set-intersection enum-set-difference
 enum-set-complement enum-set-projection

Date (srfi-19)

(current-date [tz-offset])

Note: tz-offset is currently ignored and always local time is returned

Note: If you need a JavaScript Date object, use js-eval or
(js-new "Date").

(date? x) (date-nanosecond d) (date-millisecond d)
(date-second d) (date-minute d) (date-hour d) (date-day d)
(date-month d) (date-year d) (date-week-day d)

Note: date-month returns 1..12, not 0..11

(date->string date [template])

eg. (date->string (current-date) "~Y-~m-~d ~H:~M:~S")

See srfi-19
for full list of ~?

(parse-date date) = Date.parse

Regexp

regexp-exec (regexp-exec) (regexp-replace-all)

string->regexp regexp? regexp->string

Promise (R7RS)

syntax (delay expression)

Return a promise

syntax (delay-force promise-expr)

Return a promise

promise-expr must evaluates to a promise

Mostly the same as (delay (force promise-expr)), but runs effectively

(force promise)

If no value has been computed for the promise, a value is computed and returned.

If the promise is forced a second time, the cached value is returned.

(promise? obj)

Return #t if obj is a promise

(make-promise obj)

Return a promise which, when forced, will return obj

Just return obj if obj is already a promise

Utilities

Simple format (srfi-28)

eg. (format "Result: ~s" (some-calculation))

(format format-str obj1 obj2 ...) -> string

(format #f format-str ...) -> string

(format #t format-str ...) -> output to current port

(format port format-str ...) -> output to the port

~a: display

~s: write

~%: newline

~~: tilde

Write with shared structure (srfi-38)

(write/ss x)

alias: write-with-shared-structure, write*

Syntax

Comment

Single line comment (;)

Multi line comment (#| ... |#) (srfi-30)

One sexp comment (#;) (srfi-62)

Quote

quote (')

quasiquote (`) unquote (,) unquote-splicing (,@)

JavaScript language interface

(js-eval str) evaluate str as JavaScript code

(js-ref jsobj str) = a.b

(js-set! jsobj str value) = a.b = c

(js-call jsfunc args...) = a()

(js-invoke jsobj methodname args...) = a.b()

js-invocation

macro ..

TODO: write doc

(js-new ctor args...) = new a

eg. (js-new (js-eval "Date") 2017 1 2)

(js-new ctorname args...) = new a

ctorname is evaluated as JavaScript program.

eg. (js-new "Date" 2008 1 2)

(js-obj key1 value1 key2 value2...)

eg. (js-obj "foo" 1 "bar" 2) → {"foo": 1, "bar": 2}

(js-closure proc) returns a JS function that executes a Scheme function proc

Predicates

(js-null? x)

(js-undefined? x)

(js-function? x)

Conversion

(js-array->list)

(list->js-array)

(js-obj->alist)

(alist->js-obj)

Timer

(timer proc sec) = setTimeout

(set-timer! proc sec) = setInterval

(clear-timer! timer-id) = clearInterval

(sleep sec)

Blocking sleep. (sleep 3) will wait 3 seconds and then
BiwaScheme resumes running the rest of the code.

Console

(console-debug obj1 ...) = console.debug

(console-log obj1 ...)

(console-info obj1 ...)

(console-warn obj1 ...)

(console-error obj1 ...)

Browser functions

These functions are only available in browsers (i.e. does not work on Node.js.)

Dialog

(alert msg)

= window.alert

(confirm msg) => boolean

= window.confirm

Element

(element-empty! elem) = (element-clear! elem)

(element-visible? elem)

(element-toggle! elem)

(element-hide! elem)

(element-show! elem)

(element-remove! elem)

(element-update! elem html)

(element-replace! elem x)

(element-insert! elem x)

(element-select elem)

(element-read-attribute elem attr)

(element-write-attribute! elem attr value)

(element-height elem)

(element-width elem)

(element-has-class-name? elem class)

(element-add-class-name! elem class)

(element-remove-class-name! elem class)

(element-toggle-class-name! elem class)

(element-dimensions elem) => (values width height)

(element-focus! elem)

(element-content elem) => string (html content or input value)

(element-append-child! elem child)

(element-new spec)

  (element-new '(div "foo"))        => <div>foo</div>
  (element-new '("div#main" "foo")) => <div id='main'>foo</div>
  (element-new '("div.red" "foo"))  => <div class='red'>foo</div>
  (element-new '(div align "right" "foo"))  => <div align='right'>foo</div>
  (element-new '(div (span "foo"))  => <div><span>foo</span></div>

(getelem selector) = ($ selector)

Same as jQuery(selector) except returns #f when no element is found

(set-style! selector prop value)

eg. (set-style! "#box" "left" 300)

(get-style selector prop)

(set-content! selector text)

Replace content of selector with text

Newlines in text are replaced with <br>

Tabs in text are replaced with &nbsp;&nbsp;&nbsp;

(get-content selector) => string

Event

(add-handler! selector event proc) => js-handler

(remove-handler! selector event js-handler)

eg. (define h (add-handler! "#button1" "click" (lambda (ev) ...)))

eg. (remove-handler! "#button1" "click" h)

(wait-for selector event)

eg. (wait-for "#button1" "click") (alert "pushed")

Synchronously wait for the event
(rest of the program will be evaluated after the event)

Ajax

(http-request path) => string

eg. (http-get "/foo")

Synchronously issue GET request
(rest of the program will be evaluated after server responded)

(http-post path params) => string

eg. (http-post "/foo" '(("x" . 1) ("y" . 2)))

Synchronously issue POST request
(rest of the program will be evaluated after server responded)

(receive-jsonp url) => string

eg. (receive-jsonp "http://server/x") will issue GET to
http://server/x?callback=BiwaScheme.jsonp_receiver[123]

Load Scheme or JS file on the server

(load path)

(eg. (load "lib/foo.scm"))

Synchronously load the Scheme program
(rest of the program will be evaluated after server responded)

(js-load js-path check)

(eg. (js-load "lib/foo.js" "Foo"))

Synchronously load the JavaScript file
(rest of the program will be evaluated after server responded)

Implemented with script tag. The argument check is used
for detecting the end of loading. In the above example,
BiwaScheme waits until window.Foo is defined.

Node.js

System functions

R6RS

(file-exists? filepath)

(delete-file filepath)

(command-line)

(exit) (exit num)

srfi-98

(get-environment-variable name) -> string or #f

(get-environment-variables) -> alist of string (("key" . "value"))

Loading other files

(load path)

Load a scheme source file

(js-load path)

Load a javascript source file

path is relative to the current directory, unless it starts with / or
/^\w:/ (i.e. c:, d:, etc.)

(node-require name)

Load an npm package or Node.js stdlib

eg. (node-require "fs") (node-require "marked")

Using Node.js libraries

You can also use Node.js libraries via js-eval, etc.

(define fs (js-eval "require('fs')"))

(define path ".")
(print (js-invoke fs 'readdirSync path))
; Alternatively you can use `..` macro syntax:
; (print (.. fs `(readdirSync ,path)))

BiwaScheme JavaScript API

Evaluate Scheme program

<script type="text/javascript" src="biwascheme.js"></script>
<script type="text/javascript">
var onError = function(e){ console.error(e); }
var biwa = new BiwaScheme.Interpreter(onError)
biwa.evaluate("(+ 1 2)", function(result) {
  alert(result);  //=> 3
});
</script>

Define library function

You can write JavaScript library function with BiwaScheme.define_libfunc.

1st argument: function name

2nd argument: minimum number of arguments

3rd argument: maximum number of arguments (pass null for infinite)

4th argument: function body

BiwaScheme.define_libfunc("add", 2, 2, function(ar){
  BiwaScheme.assert_integer(ar[0]);
  BiwaScheme.assert_integer(ar[1]);
  return ar[0] + ar[1];
});

See src/library/*.js for more examples.

Stringify scheme object

BiwaScheme.write(obj)

Returns a JS string whose content is the same as (write obj) generates.

Supports cyclic list/vector


ABSOLUTELY STUNNING! Just Like A Brand New Home! Many Renos Inside & Out Include: Peaked Roof, Siding, Skirting, Back Addition & Covered Deck, Siding, Sauffit, Facia, Eaves, Leveling, All Plumbing & Sewer Lines, All Electrical (Except Panel), Insulation, Drywall, Vapor Barrier, Doors, Windows, Flooring, Kitchen, Bathroom, Stainless Appliances, Front Load Washer/Dryer... All Completed in the Last Few Years. This Home has 2 Bedrooms plus a Den which is Large Enough to be a Third Bedroom. The Spacious Living Room is Open to the Upgraded Kitchen. In Between the 2 additions, There is a Covered Deck, Which Leads to the Main Door Into the Foyer Complete with Closet. There is Also a Back Door from the Deck Leading to the 2nd bedroom. Mobile City Estates is Located in the Centre of Spruce Grove & is Walking Distance to Shopping, Amenities, Walking/Bike Trails, and Schools.


Balda is a German brand founded in 1908 by Max Baldeweg in Dresden, Germany. Although not quite as legendary or well-known as many of its contemporaries like Agfa or Zeiss Ikon, Balda found their success in other ways such as manufacturing cameras like the Voigtländer Bessy AK for other companies who would then rebadge them for sale.
Camera production became a vital part of Balda in the 1920s, especially its glass plate and rollfilm cameras which continued until the outbreak of the Second World War. As part of the war effort, Balda converted its factory to manufacture gauges for the Luftwaffe (German air force) and suffered severe damage as Allied bombs rained down upon the city in February 1945, less than three months before Germany's formal surrender. As was the case with many Dresden-based camera companies, Balda split along with Germany after World War II when Max Baldeweg fled to the West German city of Bünde where he reestablished the company as Balda Kamera-Werk in 1948. The East German government seized what was left of Balda's operations in Dresden and renamed it Belca-Werk in 1951 before merging it with several other companies to form the company now known as Pentacon.
Balda Kamera-Werk continued to produce high quality consumer cameras in West Germany for sale under its own name as well as other brands until the mid 1980s. After that, camera production was moved to China in 1985 where it continued until it fizzled out sometime in the 1990s. The company then reestablished itself as Balda AG in 1999, left cameras behind, and refocused its efforts towards manufacturing medical equipment, electronic components for mobile phones, and high-precision plastics for consumer electronics, optical devices, and automobiles.
Interested in starting or growing your own collection of Balda cameras?
McKeown, James M. and Joan C. McKeown's Price Guide to Antique and Classic Cameras, 2001-2002. (Grantsburg, USA: Centennial Photo Service, 2001), 76-84.


Say goodbye to odors from cooking, pets, dirty socks, diaper pails, and musty rooms. The award winning REME HALO® in-duct air purifier is the next generation of indoor air quality (IAQ) technology and capable of purifying every cubic inch of air that your central air conditioning system reaches. It's effective against particulates, microbial, and gases. Common allergies due to mold, dander, dust, and pollen are reduced for cleaner, breathable air in your home. The system utilizes RGF's Reflective Electro Magnetic Energy technology and installs it in your HVAC system's air ducts.


November 6, 2018 November 6, 2018 Categories Uncategorizedfestival, occasion10 Comments on HAPPY DIWALI!!!!
HAPPY DIWALI TO ALL MY BELOVED READERS!!!
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Lots of thanks and beat wishes!
My pleasure cute looking girl!
By the way yr name pl!